U.S. patent application number 17/192083 was filed with the patent office on 2021-07-22 for egfr proteolysis targeting chimeric molecules and associated methods of use.
The applicant listed for this patent is Arvinas Operations, Inc., Yale University. Invention is credited to George Burslem, Andrew P. Crew, Craig M. Crews, Saul Jaime-Figueroa, Jing Wang, Kurt Zimmermann.
Application Number | 20210220475 17/192083 |
Document ID | / |
Family ID | 1000005475753 |
Filed Date | 2021-07-22 |
United States Patent
Application |
20210220475 |
Kind Code |
A1 |
Crew; Andrew P. ; et
al. |
July 22, 2021 |
EGFR PROTEOLYSIS TARGETING CHIMERIC MOLECULES AND ASSOCIATED
METHODS OF USE
Abstract
The present invention relates to bifunctional compounds, which
find utility to degrade and (inhibit) TBK1. In particular, the
present invention is directed to compounds, which contain on one
end an E3 ubiquitin ligase binding moiety which binds to an E3
ubiquitin ligase and on the other end a moiety which binds TBK1
such that TBK1 is placed in proximity to the ubiquitin ligase to
effect degradation (and inhibition) of TBK1. The present invention
exhibits a broad range of pharmacological activities associated
with compounds according to the present invention, consistent with
the degradation/inhibition of TBK1.
Inventors: |
Crew; Andrew P.; (Chester,
CT) ; Zimmermann; Kurt; (Durham, CT) ; Wang;
Jing; (Milford, CT) ; Crews; Craig M.; (New
Haven, CT) ; Jaime-Figueroa; Saul; (Morris Plains,
NJ) ; Burslem; George; (Sandwich, GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Arvinas Operations, Inc.
Yale University |
New Haven
New Haven |
CT
CT |
US
US |
|
|
Family ID: |
1000005475753 |
Appl. No.: |
17/192083 |
Filed: |
March 4, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15852854 |
Dec 22, 2017 |
10994015 |
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17192083 |
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62563494 |
Sep 26, 2017 |
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62438901 |
Dec 23, 2016 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 471/04 20130101;
A61K 31/496 20130101; A61K 45/06 20130101; A61K 31/427 20130101;
A61K 31/517 20130101; A61K 31/55 20130101; A61K 31/454 20130101;
A61K 31/551 20130101; A61K 31/52 20130101; A61K 47/55 20170801;
A61K 38/07 20130101; A61K 31/506 20130101; A61K 31/519 20130101;
A61K 47/545 20170801 |
International
Class: |
A61K 47/54 20060101
A61K047/54; A61K 47/55 20060101 A61K047/55; A61K 31/519 20060101
A61K031/519; A61K 31/427 20060101 A61K031/427; A61K 31/454 20060101
A61K031/454; A61K 31/517 20060101 A61K031/517; A61K 31/55 20060101
A61K031/55; A61K 31/496 20060101 A61K031/496; A61K 31/551 20060101
A61K031/551; A61K 45/06 20060101 A61K045/06; A61K 38/07 20060101
A61K038/07; A61K 31/52 20060101 A61K031/52; A61K 31/506 20060101
A61K031/506; C07D 471/04 20060101 C07D471/04 |
Goverment Interests
STATEMENT OF FEDERAL SUPPORT
[0002] This invention was made with government support under grant
number: R35CA197589 from the National Institutes of Health. The
government has certain rights in the invention.
Claims
1. A compound having the chemical structure: PTM-Linker-ULM, or a
pharmaceutically acceptable salt, enantiomer, stereoisomer,
solvate, polymorph or prodrug thereof, wherein: (a) the ULM is a
small molecule E3 ubiquitin ligase binding moiety that binds an E3
ubiquitin ligase selected from Von Hippel-Lindau (VLM) and cereblon
(VLM); (b) the PTM is a receptor tyrosine kinase (RTK) protein
targeting moiety having the generical structure: ##STR00813##
wherein: R.sup.k5, R.sup.k6 and R.sup.k7 are independently selected
from H, alkyl, alkyne, alkoxy, hydroxyl, cyano, halogen, haloalkyl,
haloalkoxy, and NO.sub.2, wherein the alkyl or alkoxy is optionally
substituted with 1 or 2 substituents selected from alkyl, halogen,
haloalkyl, hydroxyl, alkoxy, amino, alkylamino, dialkylamino, and
cyano; and X is N, CH, halogen, C(C.sub.1-6 alkyl), C(C.sub.1-6
haloalkyl), C(C.sub.1-6 alkoxy), C(C.sub.1-6 haloalkyl),
C(NO.sub.2), C(NH.sub.2), C(OH), or C(CN); the PTM is coupled via
the L to the ULM; and (c) the Linker is a bond or a chemical
linking moiety covalently coupling the ULM and the PTM.
2. The compound according to claim 1, wherein: R.sup.k5, R.sup.k6
and R.sup.k7 are independently selected from H, alkyl, alkyne,
alkoxy, hydroxyl, cyano, and halogen, wherein the alkyl or alkoxy
is optionally substituted with 1 to 2 substituents selected from
alkyl, halogen, haloalkyl, hydroxyl, alkoxy, amino, alkylamino,
dialkylamino and cyano; and X is N or CH.
3. The compound according to claim 1, wherein the Linker is coupled
to R.sup.k5, R.sup.k6, or R.sup.k7.
4. The compound according to claim 1, wherein the PTM is selected
from a PTM of compound 80, 82-84, 90, 91-96, 254-267, 275, 283,
289, 290, 293-300, 307, or 309-311 of FIG. 2.
5. The compound according to claim 1, wherein the ULM is a VLM
represented by: ##STR00814## wherein: X.sup.1 and X.sup.2 are each
independently selected from the group of a bond, O, NR.sup.Y3,
CR.sup.Y3R.sup.Y4, C.dbd.O, C.dbd.S, SO, and SO.sub.2; R.sup.Y3 and
R.sup.Y4 are each independently selected from the group of H,
linear or branched C.sub.1-6 alkyl, optionally substituted by 1 or
more halogen, C.sub.1-6 alkoxyl optionally substituted by 0-3
R.sup.P groups; R.sup.P is 0, 1, 2, or 3 groups, each independently
selected from the group consisting of H, halogen, --OH, C.sub.1-3
alkyl, C.dbd.O; W.sup.3 is selected from the group of an optionally
substituted -T-N(R.sup.1aR.sup.1b)X.sup.3, optionally substituted
-T-N(R.sup.1aR.sup.1b), optionally substituted -T-Aryl, an
optionally substituted -T-Heteroaryl, an optionally substituted
-T-Heterocycle, an optionally substituted --NR.sup.1-T-Aryl, an
optionally substituted --NR.sup.1-T-Heteroaryl or an optionally
substituted --NR.sup.1-T-Heterocycle; X.sup.3 is C.dbd.O, R.sup.1,
R.sup.1a, or R.sup.1b; each of R.sup.1, R.sup.1a, and R.sup.1b is
independently selected from the group consisting of H, linear or
branched C.sub.1-C.sub.6 alkyl group optionally substituted by 1 or
more halogen or --OH groups, R.sup.Y3C.dbd.O, R.sup.Y3C.dbd.S,
R.sup.Y3SO, R.sup.Y3SO.sub.2, N(R.sup.Y3R.sup.Y4)C.dbd.O,
N(R.sup.Y3R.sup.Y4)C.dbd.S, N(R.sup.Y3R.sup.Y4)SO, and
N(R.sup.Y3R.sup.Y4)SO.sub.2; T is selected from the group of an
optionally substituted alkyl, --(CH.sub.2).sub.n-- group, wherein
each one of the methylene groups is optionally substituted with one
or two substituents selected from the group of halogen, methyl, a
linear or branched C.sub.1-C.sub.6 alkyl group optionally
substituted by 1 or more halogen or --OH groups or an amino acid
side chain optionally substituted; and n is 0 to 6; W.sup.4 is
##STR00815## R.sub.14a and R.sub.14b are each independently
selected from the group of H, haloalkyl, or optionally substituted
alkyl; W.sup.5 is selected from the group of a phenyl or a 5-10
membered heteroaryl, R.sub.15 is selected from the group of H,
halogen, CN, OH, NO.sub.2, NR.sub.14aR.sub.14b, OR.sub.14a,
CONR.sub.14aR.sub.14b, NR.sub.14aCOR.sub.14b,
SO.sub.2NR.sub.14aR.sub.14b, NR.sub.14a SO.sub.2R.sub.14b,
optionally substituted alkyl, optionally substituted haloalkyl,
optionally substituted haloalkoxy; optionally substituted aryl;
optionally substituted heteroaryl; optionally substituted
cycloalkyl; or optionally substituted cycloheteroalkyl; and the
indicates the site of attachment of the PTM or the chemical linking
moiety coupling the PTM to the ULM.
6. The compound according to any of claim 1, wherein the ULM is a
VLM represented by: ##STR00816## wherein: W.sup.3 is selected from
the group of an optionally substituted aryl, optionally substituted
heteroaryl, or ##STR00817## R.sub.9 and R.sub.10 are independently
hydrogen, optionally substituted alkyl, optionally substituted
cycloalkyl, optionally substituted hydroxyalkyl, optionally
substituted heteroaryl, or haloalkyl, or R.sub.9, R.sup.10, and the
carbon atom to which they are attached form an optionally
substituted cycloalkyl; R.sub.11 is selected from the group of an
optionally substituted heterocyclic, optionally substituted alkoxy,
optionally substituted heteroaryl, optionally substituted aryl,
##STR00818## R.sub.12 is selected from the group of H or optionally
substituted alkyl; R.sub.13 is selected from the group of H,
optionally substituted alkyl, optionally substituted alkylcarbonyl,
optionally substituted (cycloalkyl)alkylcarbonyl, optionally
substituted aralkylcarbonyl, optionally substituted arylcarbonyl,
optionally substituted (heterocyclyl)carbonyl, or optionally
substituted aralkyl; R.sub.14a, R.sub.14b, are each independently
selected from the group of H, haloalkyl, or optionally substituted
alkyl; W.sup.5 is selected from the group of a phenyl or a 5-10
membered heteroaryl, R.sub.15 is selected from the group of H,
halogen, CN, OH, NO.sub.2, NR.sub.14aR.sub.14b, OR.sub.14a,
CONR.sub.14aR.sub.14b, NR.sub.14aCOR.sub.14b,
SO.sub.2NR.sub.14aR.sub.14b, NR.sub.14a SO.sub.2R.sub.14b,
optionally substituted alkyl, optionally substituted haloalkyl,
optionally substituted haloalkoxy; optionally substituted aryl;
optionally substituted heteroaryl; optionally substituted
cycloalkyl; or optionally substituted cycloheteroalkyl; R.sub.16 is
independently selected from the group of halogen, optionally
substituted alkyl, optionally substituted haloalkyl, hydroxy, or
optionally substituted haloalkoxy; o is 0, 1, 2, 3, or 4; R.sub.18
is independently selected from the group of H, halogen, optionally
substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl,
haloalkoxy or a linker; and p is 0, 1, 2, 3, or 4; and the
indicates the site of attachment of the PTM or the chemical linking
moiety coupling the PTM to the ULM.
7. The compound of any of the claim 1, wherein the ULM is a VLM
selected from the group of: ##STR00819## wherein: R.sub.1 is H,
ethyl, isopropyl, tert-butyl, sec-butyl, cyclopropyl, cyclobutyl,
cyclopentyl, or cyclohexyl; optionally substituted alkyl,
optionally substituted hydroxyalkyl, optionally substituted
heteroaryl, or haloalkyl; R.sub.14a is H, haloalkyl, optionally
substituted alkyl, methyl, fluoromethyl, hydroxymethyl, ethyl,
isopropyl, or cyclopropyl; R.sub.15 is selected from the group
consisting of H, halogen, CN, OH, NO.sub.2, optionally substituted
heteroaryl, optionally substituted thiazole, optionally substituted
imidazole, optionally substituted oxazole, optionally substituted
pyrazole, optionally substituted pyrrole, optionally substituted
furan, optionally substituted aryl; optionally substituted alkyl,
optionally substituted haloalkyl, optionally substituted
haloalkoxy, cycloalkyl, or cycloheteroalkyl; X is C or C.dbd.O;
R.sub.3 is an optionally substituted 5 or 6 membered heteroaryl;
and the indicates the site of attachment of the PTM or the chemical
linking moiety coupling the PTM to the ULM.
8. The compound according of claim 1, wherein the ULM is a VLM
represented by: wherein: ##STR00820## R.sub.14a is H, haloalkyl,
optionally substituted alkyl, methyl, fluoromethyl, hydroxymethyl,
ethyl, isopropyl, or cyclopropyl; R.sub.9 is H; R.sub.10 is H,
ethyl, isopropyl, tert-butyl, sec-butyl, cyclopropyl, cyclobutyl,
cyclopentyl, or cyclohexyl; R.sub.11 is ##STR00821## optionally
substituted heteroaryl, ##STR00822## p is 0, 1, 2, 3, or 4; and
each R.sub.18 is independently halogen, optionally substituted
alkoxy, cyano, optionally substituted alkyl, haloalkyl, or
haloalkoxy; R12 is H or C.dbd.O; R13 is H, optionally substituted
alkyl, optionally substituted alkylcarbonyl, optionally substituted
(cycloalkyl)alkylcarbonyl, optionally substituted aralkylcarbonyl,
optionally substituted arylcarbonyl, optionally substituted
(heterocyclyl)carbonyl, or optionally substituted aralkyl, R.sub.15
is selected from the group consisting of H, halogen, Cl, CN, OH,
NO.sub.2, optionally substituted heteroaryl, optionally substituted
aryl; ##STR00823## and the indicates the site of attachment of the
PTM or the chemical linking moiety coupling the PTM to the ULM.
9. The compound of claim 1, wherein the ULM is a CLM selected from
thalidomide, lenalidomide, pomalidomide, analogs thereof, isosteres
thereof, or derivatives thereof.
10. The compound of claim 1, wherein the ULM is a CLM represented
by: ##STR00824## wherein: W is selected from the group consisting
of CH.sub.2, CHR, C.dbd.O, SO.sub.2, NH, and N-alkyl; each X is
independently selected from the group consisting of absent, O, and
S; Y is selected from the group consisting of CH.sub.2,
--C.dbd.CR', NH, N-alkyl, N-aryl, N-hetaryl, N-cycloalkyl,
N-heterocyclyl, O, and S; Z is selected from the group consisting
of absent, O, and S; G and G' are independently selected from the
group consisting of H, optionally substituted alkyl, OH, R'OCOOR,
R'OCONRR'', CH.sub.2-heterocyclyl optionally substituted with R',
and benzyl optionally substituted with R'; Q.sub.1, Q.sub.2,
Q.sub.3, and Q.sub.4 represent a carbon C substituted with a group
independently selected from H, R, N or N-oxide; A is independently
selected from the group H, alkyl, cycloalkyl, Cl and F; n is an
integer from 1 to 10; R comprises --CONR'R'', --OR', --NR'R'',
--SR', --SO.sub.2R', --SO.sub.2NR'R'', --CR'R''--, --CR'NR'R''--,
(--CR'O).sub.nR'', -aryl, -hetaryl, -alkyl, -cycloalkyl,
-heterocyclyl, --P(O)(OR')R'', --P(O)R'R'', --OP(O)(OR')R'',
--OP(O)R'R'', --Cl, --F, --Br, --I, --CF.sub.3, --CN,
--NR'SO.sub.2NR'R'', --NR'CONR'R'', --CONR'COR'',
--NR'C(.dbd.N--CN)NR'R'', --C(.dbd.N--CN)NR'R'',
--NR'C(.dbd.N--CN)R'', --NR'C(.dbd.C--NO.sub.2)NR'R'',
--SO.sub.2NR'COR'', --NO.sub.2, --CO.sub.2R', --C(C.dbd.N--OR')R'',
--CR'.dbd.CR'R'', --CCR', --S(C.dbd.O)(C.dbd.N--R')R'', --SF.sub.5
and --OCF.sub.3, wherein one R is covalently joined to the linker
group (L) or to the PTM; R' and R'' are independently selected from
the group consisting of a H, optionally substituted alkyl,
cycloalkyl, aryl, heteroaryl, heterocyclic, and heterocyclyl, each
of which is optionally substituted; and represents a bond that may
be stereospecific ((R) or (S)) or non-stereospecific.
11. The compound of claim 1, wherein the ULM is a CLM represented
by: ##STR00825## wherein: W is independently selected from the
group CH.sub.2, C.dbd.O, NH, and N-alkyl; A is independently
selected from a H, methyl, and optionally substituted alkyl; n is
an integer from 1 to 4; and represents a bond that may be
stereospecific ((R) or (S)) or non-stereospecific.
12. The compound of claim 1, wherein the chemical linking moiety
comprises a chemical structural unit represented by the formula:
-(A.sup.L).sub.q-, wherein: (A.sup.L).sub.q is a group which is
connected to the ULM and the PTM; q is an integer greater than or
equal to 1; each A.sup.L is independently selected from the group
consisting of CR.sup.L1R.sup.L2, O, S, SO, SO.sub.2, NR.sup.L3,
SO.sub.2NR.sup.L3, SONR.sup.L3, CONR.sup.L3, NR.sup.L3CONR.sup.L4,
NR.sup.L3SO.sub.2NR.sup.L4, CO, CR.sup.L1.dbd.CR.sup.L2, C.ident.C,
SiR.sup.L1R.sup.L2, P(O)R.sup.L1, P(O)OR.sup.L1,
NR.sup.L3C(.dbd.NCN)NR.sup.L4, NR.sup.L3C(.dbd.NCN),
NR.sup.L3C(.dbd.CNO.sub.2)NR.sup.L4, C.sub.3-11cycloalkyl
optionally substituted with 0-6 R.sup.L1 and/or R.sup.L2 groups,
C.sub.3-11heteocyclyl optionally substituted with 1-6 R.sup.L1
and/or R.sup.L2 groups, aryl optionally substituted with 1-6
R.sup.L1 and/or R.sup.L2 groups, and heteroaryl optionally
substituted with 1-6 R.sup.L1 and/or R.sup.L2 groups, where
R.sup.L1 or R.sup.L2, each independently are optionally linked to
other groups to form cycloalkyl and/or heterocyclyl moiety,
optionally substituted with 1-4 R.sup.L5 groups; and R.sup.L1,
R.sup.L2, R.sup.L3, R.sup.L4 and R.sup.L5 are, each independently,
H, halogen, C.sub.1-8alkyl, OC.sub.1-8alkyl, SC.sub.1-8alkyl,
NHC.sub.1-8alkyl, N(C.sub.1-8alkyl).sub.2, C.sub.3-11cycloalkyl,
aryl, heteroaryl, C.sub.3-11heterocyclyl, OC.sub.3-8cycloalkyl,
SC.sub.3-8cycloalkyl, NHC.sub.3-8cycloalkyl,
N(C.sub.3-8cycloalkyl).sub.2,
N(C.sub.3-8cycloalkyl)(C.sub.1-8alkyl), OH, NH.sub.2, SH,
SO.sub.2C.sub.1-8alkyl, P(O)(OC.sub.1-8alkyl)(C.sub.1-8alkyl),
P(O)(OC.sub.1-8alkyl).sub.2, CC--C.sub.1-8alkyl, CCH,
CH.dbd.CH(C.sub.1-8alkyl),
C(C.sub.1-8alkyl).dbd.CH(C.sub.1-8alkyl),
C(C.sub.1-8alkyl).dbd.C(C.sub.1-8alkyl).sub.2, Si(OH).sub.3,
Si(C.sub.1-8alkyl).sub.3, Si(OH)(C.sub.1-8alkyl).sub.2,
COC.sub.1-8alkyl, CO.sub.2H, CN, CF.sub.3, CHF.sub.2, CH.sub.2F,
NO.sub.2, SF.sub.5, SO.sub.2NHC.sub.1-8alkyl,
SO.sub.2N(C.sub.1-8alkyl).sub.2, SONHC.sub.1-8alkyl,
SON(C.sub.1-8alkyl).sub.2, CONHC.sub.1-8alkyl,
CON(C.sub.1-8alkyl).sub.2, N(C.sub.1-8alkyl)CONH(C.sub.1-8alkyl),
N(C.sub.1-8alkyl)CON(C.sub.1-8alkyl).sub.2, NHCONH(C.sub.1-8alkyl),
NHCON(C.sub.1-8alkyl).sub.2, NHCONH.sub.2,
N(C.sub.1-8alkyl)SO.sub.2NH(C.sub.1-8alkyl), N(C.sub.1-8alkyl)
SO.sub.2N(C.sub.1-8alkyl).sub.2, NH SO.sub.2NH(C.sub.1-8alkyl), NH
SO.sub.2N(C.sub.1-8alkyl).sub.2, or NH SO.sub.2NH.sub.2.
13. The compound of claim 1, wherein the chemical linking moiety is
selected from the group consisting of:
N(R)--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2).sub.q---
O(CH2).sub.r-OCH2-,
--O--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--(CH2).sub.p--O(CH2).sub.q--O(-
CH2).sub.r-OCH2-,
--O--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2).sub.q--O-
(CH2).sub.r-O--;
--N(R)--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2).sub.q-
--O(CH2).sub.r-O--;
--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2).sub.q--O(CH-
2).sub.r-O--;
--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2).sub.q--O(CH-
2).sub.r-OCH2-; ##STR00826## ##STR00827## ##STR00828## ##STR00829##
##STR00830## ##STR00831## ##STR00832## ##STR00833## ##STR00834##
##STR00835## ##STR00836## ##STR00837## ##STR00838## ##STR00839##
##STR00840## ##STR00841## ##STR00842## ##STR00843## ##STR00844##
wherein m, n, o, p, q, and r, are independently 0, 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, with the
proviso that when m, n, o, p, q, and r is zero, there is no N--O or
O--O bond, R is selected from the group H, methyl and ethyl, and X
is selected from the group H and F.
14. The compound of claim 1, wherein the chemical linking moiety is
selected from the group consisting of: ##STR00845## ##STR00846##
##STR00847## ##STR00848## ##STR00849## ##STR00850## ##STR00851##
wherein each m and n is independently selected from 0, 1, 2, 3, 4,
5, or 6.
15. The compound of claim 1, wherein the chemical linking moiety is
selected from the group consisting of: ##STR00852## ##STR00853##
##STR00854## ##STR00855## ##STR00856## ##STR00857## ##STR00858##
##STR00859## ##STR00860## ##STR00861## ##STR00862## ##STR00863##
##STR00864## ##STR00865## ##STR00866## ##STR00867## ##STR00868##
##STR00869## ##STR00870## ##STR00871## ##STR00872## ##STR00873##
##STR00874## ##STR00875## ##STR00876## ##STR00877## ##STR00878##
##STR00879## ##STR00880## ##STR00881## ##STR00882## ##STR00883##
##STR00884## ##STR00885## ##STR00886## ##STR00887## ##STR00888##
##STR00889## ##STR00890## ##STR00891## ##STR00892## ##STR00893##
##STR00894## ##STR00895## ##STR00896## ##STR00897## ##STR00898##
##STR00899## ##STR00900## ##STR00901## ##STR00902## ##STR00903##
##STR00904## ##STR00905## wherein each m, n, o, p, q, and r is
independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
16, 17, 18, 19, or 20.
16. The compound of claim 1, wherein the chemical linking moiety is
selected from the group consisting of: ##STR00906## ##STR00907##
##STR00908## ##STR00909## ##STR00910## ##STR00911## ##STR00912##
##STR00913## ##STR00914## ##STR00915## ##STR00916## ##STR00917##
##STR00918## ##STR00919## ##STR00920## ##STR00921## ##STR00922##
##STR00923## ##STR00924##
17. The compound of claim 1, wherein the chemical linking moiety
has a chemical structure selected from: ##STR00925## wherein:
W.sup.L1 and W.sup.L2 are each independently a 4-8 membered ring
with 0-4 heteroatoms, optionally substituted with H, halogen, OH,
CN, CF.sub.3, optionally substituted linear or branched
C.sub.1-C.sub.6alkyl, optionally substituted linear or branched
C.sub.1-C.sub.6 alkoxy, or groups taken together with the atom they
are attached to, form a 4-8 membered ring system containing 0-4
heteroatoms; Y.sup.L1 is each independently a bond, optionally
substituted linear or branched C.sub.1-C.sub.6 alkyl and optionally
one or more C atoms are replaced with O; or optionally substituted
linear or branched C.sub.1-C.sub.6 alkoxy; n is 0-10; and the
##STR00926## indicates the attachment point to the PTM or the
ULM.
18. The compound of claim 1, wherein the chemical linking moiety
has a chemical structure selected from: ##STR00927## wherein:
W.sup.L1 and W.sup.L2 are each independently aryl, heteroaryl,
cyclic, heterocyclic, C.sub.1-6 alkyl, bicyclic, biaryl,
biheteroaryl, or biheterocyclic, each independently optionally
substituted with H, halogen, OH, CN, NH.sub.2, NR.sup.Y1R.sup.Y2,
CF.sub.3, hydroxyl, nitro, C.ident.CH, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl, optionally substituted linear or branched C.sub.1-C.sub.6
alkyl, optionally substituted linear or branched C.sub.1-C.sub.6
alkoxy, OC.sub.1-3alkyl optionally substituted by 1 or more --F, or
groups taken together with the atom they are attached to, form a
4-8 membered ring system containing 0-4 heteroatoms; Y.sup.L1 is
each independently a bond, NR.sup.YL1, O, S, NR.sup.YL2,
CR.sup.YL1R.sup.YL2, C.dbd.O, C.dbd.S, SO, SO.sub.2, optionally
substituted linear or branched C.sub.1-C.sub.6 alkyl and optionally
one or more C atoms are replaced with O; optionally substituted
linear or branched C.sub.1-C.sub.6 alkoxy; Q.sup.L is a 3-6
membered alicyclic or aromatic ring with 0-4 heteroatoms,
optionally bridged, optionally substituted with 0-6 R.sup.Q, each
R.sup.Q is independently H, linear or branched C.sub.1-6 alkyl
optionally substituted by 1 or more halogen or C.sub.1-6 alkoxyl,
or 2 R.sup.Q groups taken together with the atom they are attached
to, form a 3-8 membered ring system containing 0-2 heteroatoms;
R.sup.YL1, R.sup.YL2 are each independently H, OH, linear or
branched C.sub.1-6 alkyl optionally substituted by 1 or more
halogen or C.sub.1-6 alkoxyl, or R.sup.YL1, R.sup.YL2 together with
the atom they are attached to, form a 3-8 membered ring system
containing 0-2 heteroatoms; n is 0-10; and the ##STR00928##
indicates the attachment point to the PTM or the ULM.
19. The compound of claim 1, wherein the chemical linking moiety is
a polyethylenoxy group optionally substituted with aryl or phenyl
comprising from 1 to 10 ethylene glycol units.
20. A compound selected from the group consisting of examples 80,
82-84, 90, 91-96, 254-267, 275, 283, 289, 290, 293-300, 307, or
309-311 of FIG. 2.
21. A composition comprising an effective amount of a compound of
claim 1, and a pharmaceutically acceptable carrier or
excipient.
22. The composition of claim 21, wherein the composition further
comprises at least one of an additional bioactive agent or an
additional compound of claim 2.
23. The composition of claim 22, wherein the additional bioactive
agent is an anti-cancer or anti-inflammatory agent.
24. A composition comprising a pharmaceutically acceptable carrier
and an effective amount of at least one compound from examples 80,
82-84, 90, 91-96, 254-267, 275, 283, 289, 290, 293-300, 307, or
309-311 of FIG. 2.
25. A method of treating a receptor tyrosine kinase (RTK)-related
disease or disorder in a subject comprising administering to a
subject in need thereof an effective amount of a compound of claim
1 or a composition comprising the same, wherein the compound or
composition is effective for ameliorating at least one symptom of
the RTK-related disease or disorder related to EGFR overexpression
or hyperactivity, wherein the RTK-related disease or disorder is at
least one of squamous-cell carcinoma of the lung, colon cancer,
anal cancer, glioblastoma, epithelial tumor of the head and neck,
psoriasis, eczema, atherosclerosis, or a combination thereof.
26. The method of claim 25, wherein the RTK-related disease or
disorder is squamos-cell carcinoma of the lung.
27. The method of claim 25, wherein the RTK-related disease or
disorder is colon cancer.
28. The method of claim 25, wherein the RTK-related disease or
disorder is epithelial tumor of the head and neck.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a Continuation of U.S. patent
application Ser. No. 15/852,854, titled: EGFR PROTEOLYSIS TARGETING
CHIMERIC MOLECULES AND ASSOCIATED METHODS OF USE, filed 22 Dec.
2017, which claims the benefit of and priority to U.S. Provisional
Patent Application Ser. No. 62/438,901, titled: EGFR Proteolysis
Targeting Chimeric Molecules and Associated Methods of Use, filed
23 Dec. 2016, and U.S. Provisional Patent Application Ser. No.
62/563,494, titled: EGFR Proteolysis Targeting Chimeric Molecules
and Associated Methods of Use, filed 26 Sep. 2017, which are
incorporated by reference herein in their entirety.
FIELD
[0003] The description provides bifunctional compounds comprising a
target protein binding moiety and a E3 ubiquitin ligase binding
moiety, and associated methods of use. The bifunctional compounds
are useful as modulators of ubiquitination and subsequent
degradation of targeted polypeptides, in particular EGFR, which are
degraded and/or otherwise inhibited by bifunctional compounds
according to the present disclosure.
BACKGROUND
[0004] Most small molecule drugs bind enzymes or receptors in tight
and well-defined pockets. On the other hand, protein-protein
interactions are notoriously difficult to target using small
molecules due to their large contact surfaces and the shallow
grooves or flat interfaces involved. E3 ubiquitin ligases (of which
hundreds are known in humans) confer substrate specificity for
ubiquitination, and therefore, are more attractive therapeutic
targets than general proteasome inhibitors due to their specificity
for certain protein substrates. The development of ligands of E3
ligases has proven challenging, in part due to the fact that they
must disrupt protein-protein interactions. However, recent
developments have provided specific ligands which bind to these
ligases. For example, since the discovery of nutlins, the first
small molecule E3 ligase inhibitors, additional compounds have been
reported that target E3 ligases but the field remains
underdeveloped.
[0005] One E3 ligase with exciting therapeutic potential is the von
Hippel-Lindau (VHL) tumor suppressor, the substrate recognition
subunit of the E3 ligase complex VCB, which also consists of
elongins B and C, Cul2 and Rbx1. The primary substrate of VHL is
Hypoxia Inducible Factor 1.alpha. (HIF-1.alpha.), a transcription
factor that upregulates genes such as the pro-angiogenic growth
factor VEGF and the red blood cell inducing cytokine erythropoietin
in response to low oxygen levels. The first small molecule ligands
of Von Hippel Lindau (VHL) to the substrate recognition subunit of
the E3 ligase were generated, and crystal structures were obtained
confirming that the compound mimics the binding mode of the
transcription factor HIF-1.alpha., the major substrate of VHL.
[0006] Cereblon is a protein that in humans is encoded by the CRBN
gene. CRBN orthologs are highly conserved from plants to humans,
which underscores its physiological importance. Cereblon forms an
E3 ubiquitin ligase complex with damaged DNA binding protein 1
(DDB1), Cullin-4A (CUL4A), and regulator of cullins 1 (ROC1). This
complex ubiquitinates a number of other proteins. Through a
mechanism which has not been completely elucidated, cereblon
ubquitination of target proteins results in increased levels of
fibroblast growth factor 8 (FGF8) and fibroblast growth factor 10
(FGF10). FGF8 in turn regulates a number of developmental
processes, such as limb and auditory vesicle formation. The net
result is that this ubiquitin ligase complex is important for limb
outgrowth in embryos. In the absence of cereblon, DDB1 forms a
complex with DDB2 that functions as a DNA damage-binding
protein.
[0007] Inhibitors of Apotosis Proteins (IAPs) are a protein family
involved in suppressing apoptosis, i.e. cell death. The human IAP
family includes 8 members, and numerous other organisms contain IAP
homologs. IAPs contain an E3 ligase specific domain and baculoviral
IAP repeat (BIR) domains that recognize substrates, and promote
their ubiquitination. IAPs promote ubiquitination and can directly
bind and inhibit caspases. Caspases are proteases (e.g. caspase-3,
caspase-7 and caspace-9) that implement apoptosis. As such, through
the binding of caspases, IAPs inhibit cell death.
[0008] The discovery of small molecule receptor tyrosine kinase
(RTK) inhibitors greatly enabled the study of these key proteins in
normal and oncogenic signalling. Eukaryotic cell proliferation is
driven by RTK activation following binding of cognate growth
factors, and many forms of cancer are driven by the hyperactivation
of specific RTKs due either to overexpression of the protein to
super-physiological levels, or to mutations that confer growth
factor-independent activation. For example, epidermal growth factor
receptor (EGFR) is implicated in cancers and inflammatory diseases.
Activated EGFR elicits downstream activation and signaling by
several other proteins leading to DNA synthesis and cell
proliferation.
[0009] In order to obtain RTK inhibition over an extended time,
exposure to small molecule kinase inhibitors at sustained and
saturating concentrations is required. Studies have shown that
cancerous cells can co-opt other existing RTK signalling pathways
in order to permit the inhibited RTK to continue to exist as a
node, thereby restoring downstream oncogenic signalling.
[0010] Degradation of the RTK, as opposed to inhibition of the
kinase activity, is a strategy with the potential to yield a more
complete and lasting inactivation of downstream signalling and
circumvent the problem of "kinome re-wiring", whereby inhibition of
receptor signalling leads to compensatory feedback activation via
alternate kinases. Indeed, RTK elimination would prevent the
inactive kinase from persisting as a scaffolding node for oncogenic
signalling. For example, Small molecule-mediated degradation of the
protein itself rather than inhibition of the kinase domain could
provide advantages, such as reduced drug exposure time required to
suppress signalling and the ability to target kinase-independent
functions.
[0011] Bifunctional compounds such as those that are described in
U.S. Patent Application Publications 2015-0291562 and 2014-0356322
(incorporated herein by reference), function to recruit endogenous
proteins to an E3 ubiquitin ligase for degradation. In particular,
the publications describe bifunctional or proteolysis targeting
chimeric (PROTAC) compounds, which find utility as modulators of
targeted ubiquitination of a variety of polypeptides and proteins,
which are then degraded and/or otherwise inhibited by the
bifunctional compounds. Through the specific degradation of the
target proteins, the bifunctional compounds provide a therapeutic
effect.
[0012] An outstanding question, however, has been whether the
PROTAC methodology is able to induce degradation of
transmembrane-spanning proteins, given their restricted cellular
localization and the questionable accessibility of membrane-bound
receptors for ubiquitination by the cytosolic machinery. PROTACs
could provide key advantages such as improved physicochemical
properties, reduced toxicity, facile modular design, and a defined
mechanism of degradation.
[0013] There exists in the art an ongoing need for effective
treatments for disease associated with overexpression or
hyperactivation of RTK, e.g., EGFR, IGFR, and HGFR. As such,
small-molecule therapeutic agents that target RTKs are highly
desired for the treatment of RTK-related diseases.
SUMMARY
[0014] The present disclosure describes bifunctional compounds
which function to recruit endogenous RTK proteins to an E3
ubiquitin ligase for ubiquitination and subsequent degradation, and
methods of using the same. The sustained loss of RTK function can
be accomplished using Proteolysis Targeting Chimeras (PROTACs)), a
technology for post-translational protein degradation. By
chemically tethering ligands for two different proteins--an E3
ubiquitin ligase and a protein of interest--a new pharmacological
entity is created that facilitates the ubiquitination and
proteasomal degradation of the protein of interest. Its net effect
on target protein levels is similar to that achievable using RNAi
technology; however, the small-molecule approach of PROTACs does
not have the inherent liabilities of proposed nucleic-based
modalities. Indeed, PROTACs are comparable to RTK inhibitors, in
that both are amenable to adjustable dosing and can offer temporal
control to achieve the desired level of signal inactivation, nor
does it require any genetic manipulations/modification of cells in
order to work.
[0015] Thus, the present disclosure provides bifunctional
proteolysis targeting chimeric (PROTAC) compounds, which find
utility as modulators of target protein ubiquitination and
subsequent degradation. An advantage of the PROTAC compounds
provided herein is that a broad range of pharmacological activities
is possible, consistent with the degradation/inhibition of targeted
polypeptides from virtually any protein class or family. In
addition, the description provides methods of using an effective
amount of the compounds as described herein for the treatment or
amelioration of a disease condition, such as cancer.
[0016] In one aspect the disclosure provides bifunctional or PROTAC
compounds, which comprise an E3 ubiquitin ligase binding moiety
(i.e., a ligand for an E3 ubiquitin ligase or "ULM" group), and a
moiety that binds a RTK, e.g., the epidermal growth factor receptor
(EGFR) protein (i.e., a protein/polypeptide targeting moiety or
"PTM" group), such that the RTK protein is placed in proximity to
the ubiquitin ligase to effect degradation (and inhibition) of that
protein.
[0017] In a preferred embodiment, the ULM (ubiquitination ligase
modulator) can be Von Hippel-Lindau E3 ubiquitin ligase (VHL)
binding moiety (VLM), or a cereblon E3 ubiquitin ligase binding
moiety (CLM), or a mouse double minute 2 homolog (MDM2) E3
ubiquitin ligase binding moiety (MLM), or an IAP E3 ubiquitin
ligase binding moiety (i.e., a "ILM"). For example, the structure
of the bifunctional compound can be depicted as:
[PTM]-[ULM],
[0018] wherein PTM is a RTK (e.g., EGFR) binding moiety, and ULM is
an E3 ubiquitin ligase binding moiety.
[0019] The respective positions of the PTM and ULM moieties (e.g.,
VLM, CLM, MLM or ILM) as well as their number as illustrated herein
is provided by way of example only and is not intended to limit the
compounds in any way. As would be understood by the skilled
artisan, the bifunctional compounds as described herein can be
synthesized such that the number and position of the respective
functional moieties can be varied as desired.
[0020] In certain embodiments, the bifunctional compound further
comprises a chemical linker ("L"). In this example, the structure
of the bifunctional compound can be depicted as:
[PTM]-L-[ULM],
[0021] wherein PTM is a RTK (e.g., EGFR) binding moiety, L is a
linker, e.g., a bond or a chemical group coupling PTM to ULM, and
ULM is a IAP E3 ubiquitin ligase binding moiety, or a Von
Hippel-Lindau E3 ubiquitin ligase (VHL) binding moiety (VLM), or a
cereblon E3 ubiquitin ligase binding moiety (CLM), or a mouse
double minute 2 homolog (MDM2) E3 ubiquitin ligase binding moiety
(MLM).
[0022] For example, the structure of the bifunctional compound can
be depicted as:
[PTM]-L-[VLM or CLM or MLM or ILM]
[0023] wherein PTM is a RTK (e.g., EGFR) binding moiety, L is a
linker (e.g. a bond or a chemical linker group) coupling the PTM
and at least one of VLM, CLM, MLM, ILM, or a combination thereof;
VLM is Von Hippel-Lindau E3 ubiquitin ligase binding moiety that
binds to VHL E3 ligase; CLM is cereblon E3 ubiquitin ligase binding
moiety that binds to cereblon; MLM is an MDM2 E3 ubiquitin ligase
binding moiety; and ILM is a IAP binding moiety which binds to
IAP.
[0024] In certain preferred embodiments, the ILM is an AVPI
tetrapeptide fragment. As such, in certain additional embodiments,
the ILM of the bifunctional compound comprises the amino acids
alanine (A), valine (V), proline (P), and isoleucine (I) or their
unnatural mimetics, respectively. In additional embodiments, the
amino acids of the AVPI tetrapeptide fragment are connected to each
other through amide bonds (i.e., --C(O)NH-- or --NHC(O)--).
[0025] In certain embodiments, the compounds as described herein
comprise multiple independently selected ULMs, multiple PTMs,
multiple chemical linkers or a combination thereof.
[0026] In certain embodiments, ILM comprises chemical moieties such
as those described herein.
[0027] In additional embodiments, VLM can be hydroxyproline or a
derivative thereof. Furthermore, other contemplated VLMs are
included in U.S. Patent Application Publication No. 2014/03022523,
which as discussed above, is incorporated herein in its
entirety.
[0028] In an embodiment, the CLM comprises a chemical group derived
from an imide, a thioimide, an amide, or a thioamide. In a
particular embodiment, the chemical group is a phthalimido group,
or an analog or derivative thereof. In a certain embodiment, the
CLM is thalidomide, lenalidomide, pomalidomide, analogs thereof,
isosteres thereof, or derivatives thereof. Other contemplated CLMs
are described in U.S. Patent Application Publication No.
2015/0291562, which is incorporated herein in its entirety.
[0029] In certain embodiments, MLM can be nutlin or a derivative
thereof. Furthermore, other contemplated MLMs are included in U.S.
patent application Ser. No. 15/206,497 filed 11 Jul. 2016, which as
discussed above, is incorporated herein in its entirety. In certain
additional embodiments, the MLM of the bifunctional compound
comprises chemical moieties such as substituted imidazolines,
substituted spiro-indolinones, substituted pyrrolidines,
substituted piperidinones, substituted morpholinones, substituted
pyrrolopyrimidines, substituted imidazolopyridines, substituted
thiazoloimidazoline, substituted pyrrolopyrrolidinones, and
substituted isoquinolinones. In additional embodiments, the MLM
comprises the core structures mentioned above with adjacent
bis-aryl substitutions positioned as cis- or
trans-configurations.
[0030] In certain embodiments, "L" is a bond. In additional
embodiments, the linker "L" is a connector with a linear
non-hydrogen atom number in the range of 1 to 20. The connector "L"
can contain, but not limited to the functional groups such as
ether, amide, alkane, alkene, alkyne, ketone, hydroxyl, carboxylic
acid, thioether, sulfoxide, and sulfone. The linker can contain
aromatic, heteroaromatic, cyclic, bicyclic and tricyclic moieties.
Substitution with halogen, such as Cl, F, Br and I can be included
in the linker. In the case of fluorine substitution, single or
multiple fluorines can be included.
[0031] In certain embodiments, VLM is a derivative of
trans-3-hydroxyproline, where both nitrogen and carboxylic acid in
trans-3-hydroxyproline are functionalized as amides.
[0032] In certain embodiments, CLM is a derivative of
piperidine-2,6-dione, where piperidine-2,6-dione can be substituted
at the 3-position, and the 3-substitution can be bicyclic
hetero-aromatics with the linkage as C--N bond or C--C bond.
Examples of CLM can be, but not limited to, pomalidomide,
lenalidomide and thalidomide and their derivatives.
[0033] In certain embodiments, the description provides a compound
having the structure selected from compound 1-351 as described in
FIG. 2. In certain embodiments, the description provides a
therapeutic composition comprising an effective amount of at least
one compound selected from compound 1-351 as described in FIG. 2,
and a pharmaceutically acceptable carrier or excipient. In certain
embodiments, the description provides a combination for
co-administration (e.g., either separately or in a single dosage
form) comprising an effective amount of at least one compound as
described herein, at least one additional bioactive agent, and a
pharmaceutically acceptable carrier or excipient. In certain
embodiments, the additional bioactive agent is an anti-oncologic
agent.
[0034] In an additional aspect, the description provides
therapeutic compositions comprising an effective amount of a
compound as described herein or salt form thereof, and a
pharmaceutically acceptable carrier. The therapeutic compositions
modulate protein degradation in a patient or subject, for example,
an animal such as a human, and can be used for treating or
ameliorating disease states or conditions which are modulated
through the degraded protein. In certain embodiments, the
therapeutic compositions as described herein may be used to
effectuate the degradation of proteins of interest for the
treatment or amelioration of a disease, e.g., cancer.
[0035] In yet another aspect, the present disclosure provides a
method of ubiquitinating/degrading a target RTK protein, e.g.,
EGFR, in a cell. In certain embodiments, the method comprises
administering a bifunctional compound as described herein
comprising an ILM and a PTM, a PTM and a VLM, or a PTM and a CLM,
or a PTM and a MLM, preferably linked through a linker moiety, as
otherwise described herein, wherein the VLM/ILM/CLM/MLM is coupled
to the PTM through a linker to target protein that binds to PTM for
degradation. Similarly, the PTM can be coupled to VLM or CLM or MLM
or ILM through a linker to target a protein or polypeptide for
degradation. Degradation of the target protein will occur when the
target protein is placed in proximity to the E3 ubiquitin ligase,
thus resulting in degradation/inhibition of the effects of the
target protein and the control of protein levels. The control of
protein levels afforded by the present disclosure provides
treatment of a disease state or condition, which is modulated
through the target protein by lowering the level of that protein in
the cells of a patient.
[0036] In still another aspect, the description provides methods
for treating or ameliorating a disease, disorder or symptom thereof
in a subject or a patient, e.g., an animal such as a human,
comprising administering to a subject in need thereof a composition
comprising an effective amount, e.g., a therapeutically effective
amount, of a compound as described herein or salt form thereof, and
a pharmaceutically acceptable carrier, wherein the composition is
effective for treating or ameliorating the disease or disorder or
symptom thereof in the subject.
[0037] In another aspect, the description provides methods for
identifying the effects of the degradation of proteins of interest
in a biological system using compounds according to the present
disclosure.
[0038] The preceding general areas of utility are given by way of
example only and are not intended to be limiting on the scope of
the present disclosure and appended claims. Additional objects and
advantages associated with the compositions, methods, and processes
of the present disclosure will be appreciated by one of ordinary
skill in the art in light of the instant claims, description, and
examples. For example, the various aspects and embodiments of the
disclosure may be utilized in numerous combinations, all of which
are expressly contemplated by the present description. These
additional aspects and embodiments are expressly included within
the scope of the present disclosure. The publications and other
materials used herein to illuminate the background of the
invention, and in particular cases, to provide additional details
respecting the practice, are incorporated by reference.
BRIEF DESCRIPTION OF THE DRAWINGS
[0039] The accompanying drawings, which are incorporated into and
form a part of the specification, illustrate several embodiments of
the present disclosure and, together with the description, serve to
explain the principles of the invention. The drawings are only for
the purpose of illustrating an embodiment of the invention and are
not to be construed as limiting the invention. Further objects,
features and advantages of the invention will become apparent from
the following detailed description taken in conjunction with the
accompanying figures showing illustrative embodiments of the
invention, in which:
[0040] FIGS. 1A and 1B. Illustration of general principle for
PROTAC function. (1A) Exemplary PROTACs comprise a protein
targeting moiety (PTM; darkly shaded rectangle), a ubiquitin ligase
binding moiety (ULM; lightly shaded triangle), and optionally a
linker moiety (L; black line) coupling or tethering the PTM to the
ULM. (1B) Illustrates the functional use of the PROTACs as
described herein. Briefly, the ULM recognizes and binds to a
specific E3 ubiquitin ligase, and the PTM binds and recruits a
target protein bringing it into close proximity to the E3 ubiquitin
ligase. Typically, the E3 ubiquitin ligase is complexed with an E2
ubiquitin-conjugating protein, and either alone or via the E2
protein catalyzes attachment of ubiquitin (dark circles) to a
lysine on the target protein via an isopeptide bond. The
poly-ubiquitinated protein (far right) is then targeted for
degradation by the proteosomal machinery of the cell.
[0041] FIG. 2. Table of Exemplary PROTAC compounds as described
herein.
[0042] FIG. 3. Structures of exemplary PROTAC compounds as
described herein. (tPSA=total surface area). 1--Lapatinib-based
PROTAC (2 PEG linker); 2--Lapatinib-based PROTAC diastereomer (2
PEG linker); 3--Geftinib-based PROTAC; 4--Afatinib-based PROTAC;
5--Lapatinib-based PROTAC (3 PEG linker); 6--Lapatinib-based PROTAC
diastereomer (3 PEG linker); 7--Foretinib-based PROTAC;
8--Foretinib-based PROTAC diastereomer.
[0043] FIGS. 4A, 4B, 4C, 4D, 4E, 4F, and 4G. Degradation activity
of EGFR and mutants by PROTACs of FIG. 3. FIG. 4A-4F--Immunoblots
of cells expressing different EGFR variants treated with increasing
doses of the indicated compound for 24 hours. FIG. 4A--OVCAR8 cells
treated with lapatinib-based PROTAC 1. FIG. 4B--OVCAR8 cells
treated with compound 2, an inactive diastereomer of
lapatinib-based PROTAC 1. FIG. 4C--HeLa cells expressing
FLAG-tagged exon 20 insertion (ASV duplication) EGFR treated with
lapatinib-based PROTAC 1. FIG. 4D--HCC827 cells expressing exon 19
deletion EGFR treated with gefitinib-based PROTAC 2. FIG. 4E--H3255
cells expressing L858R EGFR treated with gefitinib-based PROTAC 3.
FIG. 4F--H1975 cells expressing double mutant (L858R/T790M) EGFR
treated with afatinib-based PROTAC 4. FIG. 4G--Summary table of
DC.sub.50 (the concentration at which half-maximal degradation is
achieved) and D.sub.max (the maximum percentage of degradation)
values.
[0044] FIGS. 5A, 5B, 5C, and 5D. Selective PROTAC-mediated
degradation of HER2 with the compounds of FIG. 3. FIG.
5A--Employing different linker lengths imparts PROTAC selectivity
for EGFR over HER2. OVCAR8 cells were treated with PROTAC 1 or 5
for 24 hours before being lysed and probed for EGFR, HER2 and
tubulin (as a loading control). FIG. 5B--Cell proliferation assay
in SKBr3 cells after 72 hours of treatment with the indicated
compound concentrations. FIG. 5C--Treatment of SKBr3 cells with
sub-lethal concentrations (500 nM) of PROTAC 1 or diastereomer 2
over 48 hours shows a gradual increase in downstream signalling
consistent with kinome re-wiring, previously observed in SKBr3
cells, with diastereomer but not with PROTAC. FIG.
5D--Immunoblotting analysis of c-Met phosphorylation after 48 hours
with 500 nM PROTAC 1 or diastereomer 2.
[0045] FIGS. 6A, 6B, and 6C. FIG. 6A--Gefitinib-based PROTAC 3
spares WT EGFR. OVCAR8 Cells were treated for 24 hours with
increasing doses of PROTAC 3 or with DMSO control before
immunoblotting. FIG. 6B/6C--Characterization of PROTAC 1 (6B) and
diastereomer 2 (6C) in SKBr3 cells. Cells were treated for 24 hours
in full serum with increasing doses of PROTAC 1 or with
diastereomer 2 before immunoblotting.
[0046] FIGS. 7A, 7B, 7C, 7D, 7E, and 7F. Characterization of
Foretinib-based PROTACs in GTL16 cells. FIG. 7A/7B--GTL16 cells
were treated with increasing concentrations of PROTAC 7 (7A) or
diastereomer 8 (7B) in media containing full serum for 24 hours
before immunoblotting analysis. FIG. 7C--Representative blot of
cells treated with 500 nM PROTAC 7 or 500 nM diastereomer 8 for 48
hr before immunoblotting analysis FIG. 7D--Quantification of
washout experiments. c-MET levels normalized to tubulin after
treatment with the indicated compounds at the indicated time
points. Average of 3 independent repeats and error bars represent
S.E.M. FIG. 7E--Structure of VHL-Ligand 9 used in competition
experiments. FIG. 7F--Co-treatment competition of PROTAC 7 with
VHL-Ligand 9 in MDA-MB-231 cells for 24 hours.
[0047] FIGS. 8A, 8B, 8C, 8D, and 8E. PROTAC-mediated degradation of
c-Met. FIGS. 8A and 8B--MDA-MB-231 cells treated for 24 hr with
increasing concentrations of foretinib-based PROTAC 7 (8A) or
diastereomer 8 (8B). FIGS. 8C and 8D--Cell-proliferation assay in
GTL16 cells (PROTAC 7 IC50=66.7 nM, diastereomer 8 IC50=156 nM)
(8C) and time course of c-Met degradation by foretinibbased PROTAC
(500 nM) 7 (8D). (8E) PROTAC effects are longer lasting in cell
culture. Cells were treated for 24 hr with 500 nM PROTAC 7 or
diastereomer 8 before replating on new plastic, in fresh medium for
24 or 48 hr. Excess VHL (25 mM) ligand was added to the indicated
wells.
[0048] FIGS. 9A, 9B, and 9C. PROTAC-mediated internalization. FIGS.
9A and 9B--Cell-surface proteins were labeled with a cell membrane
impermeant biotin reagent prior to treatment with 500 nM
foretinib-based PROTAC 7 (9A) or 100 ng/mL HGF (9B) for the
indicated times and lysed. Biotinylated proteins were enriched by
streptavidin pull-down and immunoblotted for c-Met. Biotinylated
proteins represent the cell-surface fraction. Corresponding
whole-cell lysates are also shown. (9C) Representative confocal
microscopy images of c-Met (green) internalization in response to
PROTAC 7 (500 nM) treatment for the indicated times (DAPI nuclear
stain in blue).
[0049] FIGS. 10A, 10B, 10C, 10D, 10E, 10F, 10G, 10H, and 10I. Exon
14-deleted c-Met has increased stability and resistance to
HGF-mediated degradation that can be combated by foretinib-based
PROTAC 7. FIG. 10A--Quantitation of WT c-Met or exon 14-deleted
c-Met degradation upon treatment with HGF, PROTAC 7, or DMSO
control in the presence of cycloheximide (CHX). FIG. 10B--Table of
calculated half-lives. FIG. 10C--Representative CHX time course of
WT c-Met degradation and signaling in MDA-MB-231 cells treated with
HGF. FIG. 10D--Representative CHX time course of exon 14-deleted
c-Met degradation and signaling in Hs746T cells treated with HGF.
FIG. 10E--Representative CHX time course of exon 14-deleted c-Met
degradation in Hs746T cells treated with PROTAC 7. FIGS. 10F and
10G--MDA-MB-231 (10F) and Hs746T (10G) cells were treated with
either DMSO or PROTAC for 18 hr before the addition of HGF and
lysis at the indicated time points following stimulation. FIG.
10H--Immunoprecipitation of c-Met from PROTAC 7-treated (1 mM) or
DMSO-treated Hs746T cells followed by immunoblotting for ubiquitin.
FIG. 10I--Tandem ubiquitin binding entity 1 (TUBE1) pull-down from
PROTAC 7 (1 mM) or DMSO-treated Hs746T cells followed by
immunoblotting for c-Met.
[0050] FIGS. 11A, 11B, 11C, and 11D. FIG. 11A--Quantitative real
time PCR was performed at the indicated timepoints after PROTAC
treatment (500 nM). Data is normalized to beta-Tubulin. FIG.
11B-11D Representative Western blots and quantitation for
cotreatment experiments. FIG. 11B--Co-treatment of PROTAC 7 (500
nM) with proteasome inhibitor epoxomicin (500 nM) for 6 hours in
MDA-MB-231 cells. Quantified data represent average of 2 repeats.
FIG. 11C--Co-treatment of PROTAC 7 (500 nM) with neddylation
inhibitor MLN-4924 (1 .mu.M) for 6 hours in MDA-MB-231 cells.
Quantified data represent average of 2 repeats. FIG.
11D--Co-treatment of PROTAC 7 (500 nM) with HSP90 inhibitor 17-AAG
(1 .mu.M) for 6 hours in MDA-MB-231 cells. Quantified data
represent average of 2 repeats.
[0051] FIGS. 12A, 12B, 12C, 12D, and 12E. Representative confocal
microscopy images of HGF-mediated internalization of c-Met. FIG.
12A--MDA-MB-231 cells treated with 100 ng/ml HGF for the indicated
times before fixing, permeabilizing, and immunostaining for c-Met.
FIG. 12B--Representative confocal microscopy images demonstrating
PROTAC-mediated colocalization with early endosome antigen 1
(EEA1). MDA-MB-231 cells treated with 500 nM PROTAC 7 for the
indicated times before fixing, permeabilizing, and immunostaining
for c-Met and EEA1. FIG. 12C--Representative confocal microscopy
images demonstrating c-Met co-localization with p230 (a trans-Golgi
marker). FIG. 12D--Quantification of images from FIG. 12C.
Percentage of cellular pixels occupied by c-Met immunofluorescence
and average cellular pixel intensity were used as a proxy for
puncta formation and reduction in cell surface c-Met. FIG.
12E--Clathrin heavy chain (CHC) siRNA experiment. MDA-MB-231 cells
were transfected with CHC siRNA before treatment with PROTAC 7 for
24 hours prior to lysis and immunoblotting.
[0052] FIGS. 13A, 13B, 13C, 13D, and 13E. Cycloheximide pulse-chase
western blots. FIG. 13A--MDA-MB-231 cells were treated with
cycloheximide followed by DMSO, PROTAC 7 or HGF and lysed at the
indicated incubation times--Set 1. FIG. 13B--MDA-MB231 cells were
treated with cycloheximide followed by DMSO, PROTAC 7 or HGF and
lysed at the indicated incubation times--Set 2. FIG. 13C--Hs746T
cells were treated with cycloheximide followed by DMSO, PROTAC 7 or
HGF and lysed at the indicated incubation times. FIG. 13D--c-Met
immunoprecipitation experiments. Hs746T cells were treated with 2
uM epoxomicin for 30 minutes before the addition of PROTAC 7 for 4
hours prior to c-Met immunoprecipitation. (WCL=Whole-cell lysate).
FIG. 13E--Hs746T cells were treated as in D prior to TUBE1
immunoprecipitation experiments.
[0053] FIG. 14. Structures of exemplary PROTAC compounds as
described herein (Lapatinib-based (furan) PROTACs).
[0054] FIGS. 15A and 15B. Degradation activity of exemplary PROTAC
compounds of FIG. 14. FIG. 15A--the percent degraded HER1 and HER2
protein at 1 uM, linker atoms, linker length (in Angstroms), linker
type and E3 ligase binding moiety (ULM) is indicated. FIG.
15B--demonstrates the degradation activity (dose-response) of HER1
in OVCAR8 cells by lapatinib-based PROTACS as indicated.
[0055] FIGS. 16A and 16B. Degradation activity of exemplary PROTAC
compounds. FIG. 16A--shows structures of exemplary lapatinib
(furan)-based PROTACs. FIG. 16B--Western blot demonstrating
degradation activity of compounds of FIG. 13A. OVCAR8 treated cells
for 24 hours. NRG (5 ng/mL) stimulation for the last 5 minutes.
Anti-EGFR rabbit (CST), anti-HER2 (Santa Cruz Biotechnologies), and
anti-tubulin (Sigma-Aldrich) were used for detection of
proteins.
[0056] FIGS. 17A and 17B. Degradation activity of exemplary PROTAC
compounds. FIG. 17A--shows structures of exemplary lapatinib
(phenyl)-based PROTACs. FIG. 17B--Western blot demonstrating
degradation activity of compounds of FIG. 17A. OVCAR8 treated cells
for 24 hours. NRG (5 ng/mL) stimulation for the last 5 minutes.
Anti-EGFR rabbit (CST), anti-HER2 (Santa Cruz Biotechnologies), and
anti-tubulin (Sigma-Aldrich) were used for detection of
proteins.
DETAILED DESCRIPTION
[0057] The following is a detailed description provided to aid
those skilled in the art in practicing the present invention. Those
of ordinary skill in the art may make modifications and variations
in the embodiments described herein without departing from the
spirit or scope of the present disclosure. All publications, patent
applications, patents, figures and other references mentioned
herein are expressly incorporated by reference in their
entirety.
[0058] Presently described are compositions and methods that relate
to the surprising and unexpected discovery that an E3 ubiquitin
ligase protein (e.g., inhibitors of apoptosis proteins (IAP), a Von
Hippel-Lindau E3 ubiquitin ligase (VHL), a cereblon E3 ubiquitin
ligase, or a mouse double minute 2 homolog (MDM2) E3 ubiquitin
ligase) ubiquitinates a target protein once it and the target
protein are placed in proximity by a bifunctional or chimeric
construct that binds the E3 ubiquitin ligase protein and the target
protein. Accordingly the present disclosure provides such compounds
and compositions comprising an E3 ubiquitin ligase binding moiety
("ULM") coupled to a protein target binding moiety ("PTM"), which
result in the ubiquitination of a chosen target protein, which
leads to degradation of the target protein by the proteasome (see
FIGS. 1A and 1B). The present disclosure also provides a library of
compositions and the use thereof.
[0059] In certain aspects, the present disclosure provides
compounds which comprise a ligand, e.g., a small molecule ligand
(i.e., having a molecular weight of below 2,000, 1,000, 500, or 200
Daltons), which is capable of binding to a ubiquitin ligase, such
as IAP, VHL, MDM2, or cereblon. The compounds also comprise a
moiety, e.g., a small molecule, that is capable of binding to
target protein, in such a way that the target protein is placed in
proximity to the ubiquitin ligase to effect degradation (and/or
inhibition) of that protein. Small molecule can mean, in addition
to the above, that the molecule is non-peptidyl, that is, it is not
generally considered a peptide, e.g., comprises fewer than 4, 3, or
2 amino acids. In accordance with the present description, the PTM,
ULM or PROTAC molecule can be a small molecule.
[0060] U.S. patent application Ser. No. 15/230,354, filed on Aug.
5, 2016; and U.S. patent application Ser. No. 15/206,497 filed 11
Jul. 2016; and U.S. patent application Ser. No. 15/209,648 filed 13
Jul. 2016; and U.S. Patent Application Ser. No. 62/406,888, filed
on Oct. 11, 2016; and U.S. patent application Ser. No. 14/686,640,
filed on Apr. 14, 2015, published as U.S. Patent Application
Publication No. 2015/0291562; and U.S. patent application Ser. No.
14/792,414, filed on Jul. 6, 2015, published as U.S. Patent
Application Publication No. 2016/0058872; and U.S. patent
application Ser. No. 14/371,956, filed on Jul. 11, 2014, published
as U.S. Patent Application Publication No. 2014/0356322; and U.S.
patent application Ser. No. 15/074,820, filed on Mar. 18, 2016,
published as U.S. Patent Application Publication No. 2016/0272639,
are incorporated herein by reference in their entirety.
Furthermore, all references cited herein are incorporated by
reference herein in their entirety.
[0061] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. The
terminology used in the description is for describing particular
embodiments only and is not intended to be limiting of the
invention.
[0062] Where a range of values is provided, it is understood that
each intervening value, to the tenth of the unit of the lower limit
unless the context clearly dictates otherwise (such as in the case
of a group containing a number of carbon atoms in which case each
carbon atom number falling within the range is provided), between
the upper and lower limit of that range and any other stated or
intervening value in that stated range is encompassed within the
invention. The upper and lower limits of these smaller ranges may
independently be included in the smaller ranges is also encompassed
within the invention, subject to any specifically excluded limit in
the stated range. Where the stated range includes one or both of
the limits, ranges excluding either both of those included limits
are also included in the invention.
[0063] The following terms are used to describe the present
invention. In instances where a term is not specifically defined
herein, that term is given an art-recognized meaning by those of
ordinary skill applying that term in context to its use in
describing the present invention.
[0064] The articles "a" and "an" as used herein and in the appended
claims are used herein to refer to one or to more than one (i.e.,
to at least one) of the grammatical object of the article unless
the context clearly indicates otherwise. By way of example, "an
element" means one element or more than one element.
[0065] The phrase "and/or," as used herein in the specification and
in the claims, should be understood to mean "either or both" of the
elements so conjoined, i.e., elements that are conjunctively
present in some cases and disjunctively present in other cases.
Multiple elements listed with "and/or" should be construed in the
same fashion, i.e., "one or more" of the elements so conjoined.
Other elements may optionally be present other than the elements
specifically identified by the "and/or" clause, whether related or
unrelated to those elements specifically identified. Thus, as a
non-limiting example, a reference to "A and/or B", when used in
conjunction with open-ended language such as "comprising" can
refer, in one embodiment, to A only (optionally including elements
other than B); in another embodiment, to B only (optionally
including elements other than A); in yet another embodiment, to
both A and B (optionally including other elements); etc.
[0066] As used herein in the specification and in the claims, "or"
should be understood to have the same meaning as "and/or" as
defined above. For example, when separating items in a list, "or"
or "and/or" shall be interpreted as being inclusive, i.e., the
inclusion of at least one, but also including more than one, of a
number or list of elements, and, optionally, additional unlisted
items. Only terms clearly indicated to the contrary, such as "only
one of" or "exactly one of," or, when used in the claims,
"consisting of," will refer to the inclusion of exactly one element
of a number or list of elements. In general, the term "or" as used
herein shall only be interpreted as indicating exclusive
alternatives (i.e., "one or the other but not both") when preceded
by terms of exclusivity, such as "either," "one of," "only one of,"
or "exactly one of."
[0067] In the claims, as well as in the specification above, all
transitional phrases such as "comprising," "including," "carrying,"
"having," "containing," "involving," "holding," "composed of," and
the like are to be understood to be open-ended, i.e., to mean
including but not limited to. Only the transitional phrases
"consisting of" and "consisting essentially of" shall be closed or
semi-closed transitional phrases, respectively, as set forth in the
United States Patent Office Manual of Patent Examining Procedures,
Section 2111.03.
[0068] As used herein in the specification and in the claims, the
phrase "at least one," in reference to a list of one or more
elements, should be understood to mean at least one element
selected from anyone or more of the elements in the list of
elements, but not necessarily including at least one of each and
every element specifically listed within the list of elements and
not excluding any combinations of elements in the list of elements.
This definition also allows that elements may optionally be present
other than the elements specifically identified within the list of
elements to which the phrase "at least one" refers, whether related
or unrelated to those elements specifically identified. Thus, as a
nonlimiting example, "at least one of A and B" (or, equivalently,
"at least one of A or B," or, equivalently "at least one of A
and/or B") can refer, in one embodiment, to at least one,
optionally including more than one, A, with no B present (and
optionally including elements other than B); in another embodiment,
to at least one, optionally including more than one, B, with no A
present (and optionally including elements other than A); in yet
another embodiment, to at least one, optionally including more than
one, A, and at least one, optionally including more than one, B
(and optionally including other elements); etc.
[0069] It should also be understood that, in certain methods
described herein that include more than one step or act, the order
of the steps or acts of the method is not necessarily limited to
the order in which the steps or acts of the method are recited
unless the context indicates otherwise.
[0070] The terms "co-administration" and "co-administering" or
"combination therapy" refer to both concurrent administration
(administration of two or more therapeutic agents at the same time)
and time varied administration (administration of one or more
therapeutic agents at a time different from that of the
administration of an additional therapeutic agent or agents), as
long as the therapeutic agents are present in the patient to some
extent, preferably at effective amounts, at the same time. In
certain preferred aspects, one or more of the present compounds
described herein, are coadministered in combination with at least
one additional bioactive agent, especially including an anticancer
agent. In particularly preferred aspects, the co-administration of
compounds results in synergistic activity and/or therapy, including
anticancer activity.
[0071] The term "compound", as used herein, unless otherwise
indicated, refers to any specific chemical compound disclosed
herein and includes tautomers, regioisomers, geometric isomers, and
where applicable, stereoisomers, including optical isomers
(enantiomers) and other stereoisomers (diastereomers) thereof, as
well as pharmaceutically acceptable salts and derivatives,
including prodrug and/or deuterated forms thereof where applicable,
in context. Deuterated small molecules contemplated are those in
which one or more of the hydrogen atoms contained in the drug
molecule have been replaced by deuterium.
[0072] Within its use in context, the term compound generally
refers to a single compound, but also may include other compounds
such as stereoisomers, regioisomers and/or optical isomers
(including racemic mixtures) as well as specific enantiomers or
enantiomerically enriched mixtures of disclosed compounds. The term
also refers, in context to prodrug forms of compounds which have
been modified to facilitate the administration and delivery of
compounds to a site of activity. It is noted that in describing the
present compounds, numerous substituents and variables associated
with same, among others, are described. It is understood by those
of ordinary skill that molecules which are described herein are
stable compounds as generally described hereunder. When the bond is
shown, both a double bond and single bond are represented or
understood within the context of the compound shown and well-known
rules for valence interactions.
[0073] The term "ubiquitin ligase" refers to a family of proteins
that facilitate the transfer of ubiquitin to a specific substrate
protein, targeting the substrate protein for degradation. For
example, IAP an E3 ubiquitin ligase protein that alone or in
combination with an E2 ubiquitin-conjugating enzyme causes the
attachment of ubiquitin to a lysine on a target protein, and
subsequently targets the specific protein substrates for
degradation by the proteasome. Thus, E3 ubiquitin ligase alone or
in complex with an E2 ubiquitin conjugating enzyme is responsible
for the transfer of ubiquitin to targeted proteins. In general, the
ubiquitin ligase is involved in polyubiquitination such that a
second ubiquitin is attached to the first; a third is attached to
the second, and so forth. Polyubiquitination marks proteins for
degradation by the proteasome. However, there are some
ubiquitination events that are limited to mono-ubiquitination, in
which only a single ubiquitin is added by the ubiquitin ligase to a
substrate molecule. Mono-ubiquitinated proteins are not targeted to
the proteasome for degradation, but may instead be altered in their
cellular location or function, for example, via binding other
proteins that have domains capable of binding ubiquitin. Further
complicating matters, different lysines on ubiquitin can be
targeted by an E3 to make chains. The most common lysine is Lys48
on the ubiquitin chain. This is the lysine used to make
polyubiquitin, which is recognized by the proteasome.
[0074] The term "patient" or "subject" is used throughout the
specification to describe an animal, preferably a human or a
domesticated animal, to whom treatment, including prophylactic
treatment, with the compositions according to the present
disclosure is provided. For treatment of those infections,
conditions or disease states which are specific for a specific
animal such as a human patient, the term patient refers to that
specific animal, including a domesticated animal such as a dog or
cat or a farm animal such as a horse, cow, sheep, etc. In general,
in the present disclosure, the term patient refers to a human
patient unless otherwise stated or implied from the context of the
use of the term.
[0075] The term "effective" is used to describe an amount of a
compound, composition or component which, when used within the
context of its intended use, effects an intended result. The term
effective subsumes all other effective amount or effective
concentration terms, which are otherwise described or used in the
present application.
[0076] As some of the most compelling anti-cancer targets are RTKs,
the demonstration of their susceptibility to PROTAC-mediated
degradation has remained an important question. Given their
well-defined role in human cancers and the broad understanding of
their regulation and downstream signalling pathways, EGFR, HER2 and
c-Met represent potential PROTAC targets of interest.
[0077] Herein, we show effective PROTAC-mediated degradation of
these RTKs, including a number of relevant oncogenic mutant
isoforms. The described results demonstrate that not only are RTKs
viable substrates for post-translational degradation, but also that
the signalling inactivation and growth inhibition achieved by
PROTACs is more potent, more sustained, and less susceptible to
kinome re-wiring than that achieved via RTK inhibition.
[0078] Compounds and Compositions
[0079] In one aspect, the description provides compounds comprising
an E3 ubiquitin ligase binding moiety ("ULM") that is an IAP E3
ubiquitin ligase binding moiety (an "ILM"), a cereblon E3 ubiquitin
ligase binding moiety (a "CLM"), a Von Hippel-Lindae E3 ubiquitin
ligase (VHL) binding moiety (VLM), and/or a mouse double minute 2
homologue (MDM2) E3 ubiquitin ligase binding moiety (MLM). In an
exemplary embodiment, the ULM is coupled to a RTK target protein
binding moiety (PTM) via a chemical linker (L) according to the
structure:
PTM-L-ULM (A)
wherein L is a bond or a chemical linker group, ULM is a E3
ubiquitin ligase binding moiety, and PTM is a target protein
binding moiety. The number and/or relative positions of the
moieties in the compounds illustrated herein is provided by way of
example only. As would be understood by the skilled artisan,
compounds described herein can be synthesized with any desired
number and/or relative position of the respective functional
moieties.
[0080] The terms ULM, ILM, VLM, MLM, and CLM are used in their
inclusive sense unless the context indicates otherwise. For
example, the term ULM is inclusive of all ULMs, including those
that bind IAP (i.e., ILMs), MDM2 (i.e., MLM), cereblon (i.e., CLM),
and VHL (i.e., VLM). Further, the term ILM is inclusive of all
possible IAP E3 ubiquitin ligase binding moieties, the term MLM is
inclusive of all possible MDM2 E3 ubiquitin ligase binding
moieties, the term VLM is inclusive of all possible VHL binding
moieties, and the term CLM is inclusive of all cereblon binding
moieties.
[0081] In another aspect, the present disclosure provides
bifunctional or multifunctional compounds (e.g., PROTACs) useful
for regulating protein activity by inducing the degradation of a
target protein. In certain embodiments, the compound comprises an
ILM or a VLM or a CLM or a MLM coupled, e.g., linked covalently,
directly or indirectly, to a moiety that binds a target protein
(i.e., a protein targeting moiety or a "PTM"). In certain
embodiments, the ILM/VLM/CLM/MLM and PTM are joined or coupled via
a chemical linker (L). The ILM binds the IAP E3 ubiquitin ligase,
the VLM binds VHL, CLM binds the cereblon E3 ubiquitin ligase, and
MLM binds the MDM2 E3 ubiquitin ligase, and the PTM recognizes a
target protein and the interaction of the respective moieties with
their targets facilitates the degradation of the target protein by
placing the target protein in proximity to the ubiquitin ligase
protein. An exemplary bifunctional compound can be depicted as:
PTM-ILM (B)
PTM-CLM (C)
PTM-VLM (D)
PTM-MLM (E)
[0082] In certain embodiments, the bifunctional compound further
comprises a chemical linker ("L"). For example, the bifunctional
compound can be depicted as:
PTM-L-ILM (F)
PTM-L-CLM (G)
PTM-L-VLM (H)
PTM-L-MLM (I)
[0083] wherein the PTM is a protein/polypeptide targeting moiety,
the L is a chemical linker, the ILM is a IAP E3 ubiquitin ligase
binding moiety, the CLM is a cereblon E3 ubiquitin ligase binding
moiety, the VLM is a VHL binding moiety, and the MLM is a MDM2 E3
ubiquitin ligase binding moiety.
[0084] In certain embodiments, the ULM (e.g., a ILM, a CLM, a VLM,
or a MLM) shows activity or binds to the E3 ubiquitin ligase (e.g.,
IAP E3 ubiquitin ligase, cereblon E3 ubiquitin ligase, VHL, or MDM2
E3 ubiquitin ligase) with an IC.sub.50 of less than about 200
.mu.M. The IC.sub.50 can be determined according to any method
known in the art, e.g., a fluorescent polarization assay.
[0085] In certain additional embodiments, the bifunctional
compounds described herein demonstrate an activity with an
IC.sub.50 of less than about 100, 50, 10, 1, 0.5, 0.1, 0.05, 0.01,
0.005, 0.001 mM, or less than about 100, 50, 10, 1, 0.5, 0.1, 0.05,
0.01, 0.005, 0.001 .mu.M, or less than about 100, 50, 10, 1, 0.5,
0.1, 0.05, 0.01, 0.005, 0.001 nM, or less than about 100, 50, 10,
1, 0.5, 0.1, 0.05, 0.01, 0.005, 0.001 .mu.M.
[0086] In certain embodiments, the compounds as described herein
comprise multiple PTMs (targeting the same or different protein
targets), multiple ULMs, one or more ULMs (i.e., moieties that bind
specifically to multiple/different E3 ubiquitin ligase, e.g., VHL,
IAP, cereblon, and/or MDM2) or a combination thereof. In any of the
aspects of embodiments described herein, the PTMs and ULMs (e.g.,
ILM, VLM, CLM, and/or MLM) can be coupled directly or via one or
more chemical linkers or a combination thereof. In additional
embodiments, where a compound has multiple ULMs, the ULMs can be
for the same E3 ubiquitin ligase or each respective ULM can bind
specifically to a different E3 ubiquitin ligase. In still further
embodiments, where a compound has multiple PTMs, the PTMs can bind
the same target protein or each respective PTM can bind
specifically to a different target protein.
[0087] In certain embodiments, where the compound comprises
multiple ULMs, the ULMs are identical. In additional embodiments,
the compound comprising a plurality of ULMs (e.g., ULM, ULM',
etc.), at least one PTM coupled to a ULM directly or via a chemical
linker (L) or both. In certain additional embodiments, the compound
comprising a plurality of ULMs further comprises multiple PTMs. In
still additional embodiments, the PTMs are the same or, optionally,
different. In still further embodiments, wherein the PTMs are
different, the respective PTMs may bind the same protein target or
bind specifically to a different protein target.
[0088] In certain embodiments, the compound may comprise a
plurality of ULMs and/or a plurality of ULM's. In further
embodiments, the compound comprising at least two different ULMs, a
plurality of ULMs, and/or a plurality of ULM's further comprises at
least one PTM coupled to a ULM or a ULM' directly or via a chemical
linker or both. In any of the embodiments described herein, a
compound comprising at least two different ILMs can further
comprise multiple PTMs. In still additional embodiments, the PTMs
are the same or, optionally, different. In still further
embodiments, wherein the PTMs are different the respective PTMs may
bind the same protein target or bind specifically to a different
protein target. In still further embodiments, the PTM itself is a
ULM (or ULM'), such as an ILM, a VLM, a CLM, a MLM, an ILM', a
VLM', a CLM', and/or a MLM'.
[0089] In additional embodiments, the description provides the
compounds as described herein including their enantiomers,
diastereomers, solvates and polymorphs, including pharmaceutically
acceptable salt forms thereof, e.g., acid and base salt forms.
[0090] Exemplary ILMs
[0091] AVPI Tetrapeptide Fragments
[0092] In any of the compounds described herein, the ILM can
comprise an alanine-valine-proline-isoleucine (AVPI) tetrapeptide
fragment or an unnatural mimetic thereof. In certain embodiments,
the ILM is selected from the group consisting of chemical
structures represented by Formulas (I), (II), (III), (IV), and
(V):
##STR00001##
wherein: [0093] R.sup.1 for Formulas (I), (II), (III), (IV), and
(V) is selected from H or alkyl; [0094] R.sup.2 for Formulas (I),
(II), (III), (IV), and (V) is selected from H or alkyl; [0095]
R.sup.3 for Formulas (I), (II), (III), (IV), and (V) is selected
from H, alkyl, cycloalkyl and heterocycloalkyl; [0096] R.sup.5 and
R.sup.6 for Formulas (I), (II), (III), (IV), and (V) are
independently selected from H, alkyl, cycloalkyl, heterocycloalkyl,
or more preferably, R.sup.5 and R.sup.6 taken together for Formulas
(I), (II), (III), (IV), and (V) form a pyrrolidine or a piperidine
ring further optionally fused to 1-2 cycloalkyl, heterocycloalkyl,
aryl or heteroaryl rings, each of which can then be further fused
to another cycloalkyl, heterocycloalkyl, aryl or heteroaryl ring;
[0097] R.sup.3 and R.sup.5 for Formulas (I), (II), (III), (IV), and
(V) taken together can form a 5-8-membered ring further optionally
fused to 1-2 cycloalkyl, heterocycloalkyl, aryl or heteroaryl
rings; [0098] R.sup.7 for Formulas (I), (II), (III), (IV), and (V)
is selected from cycloalkyl, cycloalkylalkyl, heterocycloalkyl,
heterocycloalkylalkyl, aryl, arylalkyl, heteroaryl or
heteroarylalkyl, each one further optionally substituted with 1-3
substituents selected from halogen, alkyl, haloalkyl, hydroxyl,
alkoxy, cyano, (hetero)cycloalkyl or (hetero)aryl, or R.sup.7 is
--C(O)NH--R.sup.4; and [0099] R.sup.4 is selected from alkyl,
cycloalkyl, heterocycloalkyl, cycloalkylalkyl,
heterocycloalkylalkyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl, further optionally substituted with 1-3
substituents as described above.
[0100] As shown above, P1, P2, P3, and P4 of Formula (II) correlate
with A, V, P, and I, respectively, of the AVPI tetrapeptide
fragment or an unnatural mimetic thereof. Similarly, each of
Formulas (I) and (III) through (V) have portions correlating with
A, V, P, and I of the AVPI tetrapeptide fragment or an unnatural
mimetic thereof.
[0101] In any of the compounds described herein, the ILM can have
the structure of Formula (VI), which is a derivative of IAP
antagonists described in WO Pub. No. 2008/014236, or an unnatural
mimetic thereof:
##STR00002##
wherein: [0102] R.sub.1 of Formula (VI) is, independently selected
from H C.sub.1-C.sub.4-alky, C.sub.1-C.sub.4-alkenyl,
C.sub.1-C.sub.4-alkynyl or C.sub.3-C.sub.10-cycloalkyl which are
unsubstituted or substituted; [0103] R.sub.2 of Formula (VI) is,
independently selected from H, C.sub.1-C.sub.4-alkyl,
C.sub.1-C.sub.4-alkenyl, C.sub.1-C.sub.4-alkynyl or
C.sub.3-C.sub.10-cycloalkyl which are unsubstituted or substituted;
[0104] R.sub.3 of Formula (VI) is, independently selected from H,
--CF.sub.3, --C.sub.2H.sub.5, C.sub.1-C.sub.4-alkyl,
C.sub.1-C.sub.4-alkenyl, C.sub.1-C.sub.4-alkynyl, --CH.sub.2--Z or
any R.sub.2 and R.sub.3 together form a heterocyclic ring: each Z
of Formula (VI) is, independently selected from H, --OH, F, Cl,
--CH.sub.3, --CF.sub.3, --CH.sub.2Cl, --CH.sub.2F or --CH.sub.2OH;
[0105] R.sub.4 of Formula (VI) is, independently selected from
C.sub.1-C.sub.16 is straight or branched alkyl,
C.sub.1-C.sub.16-alkenyl, C.sub.1-C.sub.16-alkynyl,
C.sub.3-C.sub.10-cycloalkyl, --(CH.sub.2).sub.0-6--Z.sub.1,
--(CH.sub.2).sub.0-6-aryl, and --(CH.sub.2).sub.0-6-het, wherein
alkyl, cycloalkyl, and phenyl are unsubstituted or substituted;
[0106] R.sub.5 of Formula (VI) is, independently selected from H,
C.sub.1-10-alkyl, aryl, phenyl, C.sub.3-7-cycloalkyl,
--(CH.sub.2).sub.1-6--C.sub.3-7-cycloalkyl,
--C.sub.1-10-alkyl-aryl,
--(CH.sub.2).sub.0-6--C.sub.3-7-cycloalkyl-(CH.sub.2).sub.0-6-phenyl,
--(CH.sub.2).sub.0-4--CH[(CH.sub.2).sub.1-4-phenyl].sub.2, indanyl,
--C(O)--C.sub.1-10-alkyl,
--C(O)--(CH.sub.2).sub.1-6--C.sub.3-7-cycloalkyl,
--C(O)--(CH.sub.2).sub.0-6-phenyl,
--(CH.sub.2).sub.0-6--C(O)-phenyl, --(CH.sub.2).sub.0-6-het,
--C(O)--(CH.sub.2).sub.1-6-het, or R.sub.5 is selected from a
residue of an amino acid, wherein the alkyl, cycloalkyl, phenyl,
and aryl substituents are unsubstituted or substituted; [0107]
Z.sub.1 of Formula (VI) is, independently selected from
--N(R.sub.10)--C(O)--C.sub.1-10-alkyl,
--N(R.sub.10)--C(O)--(CH.sub.2).sub.0-6--C.sub.3-7-cycloalkyl,
--N(R.sub.10)--C(O)--(CH.sub.2).sub.0-6-phenyl,
--N(R.sub.10)--C(O)(CH.sub.2).sub.1-6-het,
--C(O)--N(R.sub.11)(R.sub.12), --C(O)--O--C.sub.1-10-alkyl,
--C(O)--O--(CH.sub.2).sub.1-6--C.sub.3-7-cycloalkyl,
--C(O)--O--(CH.sub.2).sub.0-6-phenyl,
--C(O)--O--(CH.sub.2).sub.1-6-het, --O--C(O)--C.sub.1-10-alkyl,
--O--C(O)--(CH.sub.2).sub.1-6--C.sub.3-7-cycloalkyl,
--O--C(O)--(CH.sub.2).sub.0-66-phenyl,
--O--C(O)--(CH.sub.2).sub.1-6-het, wherein alkyl, cycloalkyl, and
phenyl are unsubstituted or substituted; [0108] het of Formula (VI)
is, independently selected from a 5-7 member heterocyclic ring
containing 1-4 heteroatoms selected from N, O, and S, or an 8-12
member fused ring system including at least one 5-7 member
heterocyclic ring containing 1, 2, or 3 heteroatoms selected from
N, O, and S, which heterocyclic ring or fused ring system is
unsubstituted or substituted on a carbon or nitrogen atom; [0109]
R.sub.10 of Formula (VI) is selected from H, --CH.sub.3,
--CF.sub.3, --CH.sub.2OH, or --CH.sub.2Cl; [0110] R.sub.11 and
R.sub.12 of Formula (VI) are independently selected from H,
C.sub.1-4-alkyl, C.sub.3-7-cycloalkyl,
--(CH.sub.2).sub.1-6--C.sub.3-7-cycloakyl,
(CH.sub.2).sub.0-6-phenyl, wherein alkyl, cycloalkyl, and phenyl
are unsubstituted or substituted; or R.sub.11 and R.sub.12 together
with the nitrogen form het, and U of Formula (VI) is,
independently, as shown in Formula (VII):
##STR00003##
[0110] wherein: [0111] each n of Formula (VII) is, independently
selected from 0 to 5; [0112] X of Formula (VII) is selected from
the group --CH and N; [0113] R.sub.a and R.sub.b, of Formula (VII)
are independently selected from the group O, S, or N atom or
C.sub.0-8-alkyl wherein one or more of the carbon atoms in the
alkyl chain are optionally replaced by a heteroatom selected from
O, S, or N, and where each alkyl is, independently, either
unsubstituted or substituted; [0114] R.sub.d of Formula (VII) is
selected from the group Re-Q-(R.sub.f).sub.p(R.sub.g).sub.q, and
Ar.sub.1-D-Ar.sub.2; [0115] R.sub.c of Formula (VII) is selected
from the group H or any R.sub.c and R.sub.d together form a
cycloalkyl or het; where if R.sub.c and R.sub.d form a cycloalkyl
or het, R.sub.5 is attached to the formed ring at a C or N atom:
[0116] p and q of Formula (VII) are independently selected from 0
or 1; [0117] R.sub.e of Formula (VII) is selected from the group
C.sub.1-8-alkyl and alkylidene, and each Re is either unsubstituted
or substituted; [0118] Q is selected from the group N, O, S, S(O),
and S(O).sub.2; [0119] Ar.sub.1 and Ar.sub.2 of Formula (VII) are
independently selected from the group of substituted or
unsubstituted aryl and het; [0120] R.sub.f and R.sub.g of Formula
(VII) are independently selected from H, --C1-10-alkyl,
C.sub.1-10-alkylaryl, --OH, --O--C.sub.1-10-alkyl,
--(CH.sub.2).sub.0-6--C.sub.3-7-cycloalky,
--O--(CH.sub.2).sub.0-6-aryl, phenyl, aryl, phenyl-phenyl,
--(CH.sub.2).sub.1-6-het, --O--(CH.sub.2).sub.1-6-het, --OR.sub.13,
--C(0)-R.sub.13, --C(O)--N(R.sub.13)(R.sub.14),
--N(R.sub.13)(R.sub.14), --S--R.sub.13, --S(O)--R.sub.13,
--S(O).sub.2--R.sub.13, --S(O).sub.2--NR.sub.13R.sub.14,
--NR.sub.13--S(O).sub.2--R.sub.14, --S--C.sub.1-10-alkyl,
aryl-C.sub.1-4-alkyl, or het-C.sub.1-4-alkyl wherein alkyl,
cycloalkyl, het, and aryl are unsubstituted or substituted,
--SO.sub.2--C.sub.1-2-alkyl, --SO.sub.2--C.sub.1-2-alkylphenyl,
--O--C.sub.1-4-alkyl, or any R.sub.g and R.sub.f together form a
ring selected from het or aryl; [0121] D of Formula (VII) is
selected from the group --CO--, --C(O)--C.sub.1-7-alkylene or
arylene, --CF.sub.2--, --O--, --S(O).sub.r where r is 0-2,
1,3-dioxalane, or C.sub.1-7-alkyl-OH; where alkyl, alkylene, or
arylene are unsubstituted or substituted with one or more halogens,
OH, --O--C.sub.1-6-alkyl, --S--C.sub.1-6-alkyl, or --CF.sub.3; or
each D is, independently selected from N(R.sub.h); [0122] Rh is
selected from the group H, unsubstituted or substituted
C.sub.1-7-alkyl, aryl, unsubstituted or substituted
--O--(C.sub.1-7-cycloalkyl), --C(O)--C.sub.1-10-alkyl,
--C(O)--C.sub.0-10-alkyl-aryl, --C--O--C.sub.01-10-alkyl,
--C--O--C.sub.0-10-alkyl-aryl, --SO.sub.2--C.sub.1-10-alkyl, or
--SO.sub.2--(C.sub.0-10-alkylaryl); [0123] R.sub.6, R.sub.7,
R.sub.8, and R.sub.9 of Formula (VII) are, independently, selected
from the group H, --C.sub.1-10-alkyl, --C.sub.1-10-alkoxy,
aryl-C.sub.1-10-alkoxy, --OH, --O--C.sub.1-10-alkyl,
--(CH.sub.2).sub.0-6--C.sub.3-7-cycloalkyl,
--O--(CH.sub.2).sub.0-6-aryl, phenyl, --(CH.sub.2).sub.1-6-het,
--O--(CH.sub.2).sub.1-6-het, --OR.sub.13, --C(O)--R.sub.13,
--C(O)--N(R.sub.13)(R.sub.14), --N(R.sub.13)(R.sub.14),
--S--R.sub.13, --S(O)--R.sub.13, --S(O).sub.2--R.sub.13,
--S(O).sub.2--NR.sub.13R.sub.14, or
--NR.sub.13--S(O).sub.2--R.sub.14; wherein each alkyl, cycloalkyl,
and aryl is unsubstituted or substituted and any R.sub.6, R.sub.7,
R.sub.8, and R9 optionally together form a ring system [0124]
R.sub.13 and R.sub.14 of Formula (VII) are independently selected
from the group H, C.sub.1-10-alkyl,
--(CH.sub.2).sub.0-6--C.sub.3-7-cycloalkyl,
--(CH.sub.2).sub.0-6--(CH).sub.0-1-(aryl).sub.1-2,
--C(O)--C.sub.1-10-alkyl,
--C(O)--(CH.sub.2).sub.1-6--C.sub.3-7-cycloalkyl,
--C(O)--O--(CH.sub.2).sub.0-6-aryl,
--C(O)--(CH.sub.2).sub.0-6--O-fluorenyl,
--C(O)--NH--(CH.sub.2).sub.0-6-aryl,
--C(O)--(CH.sub.2).sub.0-6-aryl, --C(O)--(CH.sub.2).sub.0-6-het,
--C(S)--C.sub.1-10-alkyl,
--C(S)--(CH.sub.2).sub.1-6--C.sub.3-7-cycloalkyl,
--C(S)--O--(CH.sub.2).sub.0-6-aryl,
--C(S)--(CH.sub.2).sub.0-6--O-fluorenyl,
--C(S)--NH--(CH.sub.2).sub.0-6-aryl,
--C(S)--(CH.sub.2).sub.0-6-aryl, or --C(S)--(CH.sub.2).sub.1-6-het,
wherein each alkyl, cycloalkyl and aryl is unsubstituted or
substituted: or any R.sub.13 and R.sub.14 together with a nitrogen
atom form het; [0125] wherein alkyl substituents of R.sub.13 and
R.sub.14 of Formula (VII) are unsubstituted or substituted and when
substituted, are substituted by one or more substituents selected
from C.sub.1-10 alkyl, halogen, OH, --O--C.sub.1-6-alkyl,
--S--C.sub.1-6-alkyl, and --CF.sub.3; and substituted phenyl or
aryl of R.sub.13 and R.sub.14 are substituted by one or more
substituents selected from halogen, hydroxyl. C.sub.1-4-alkyl,
C.sub.1-4-alkoxy, nitro, --CN, --O--C(O)--C.sub.1-4-alkyl, and
--C(O)--O--C.sub.1-4-aryl; or a pharmaceutically acceptable salt or
hydrate thereof.
[0126] In certain embodiments, the compound further comprises an
independently selected second ILM attached to the ILM of Formula
(VI), or an unnatural mimetic thereof, by way of at least one
additional independently selected linker group. In an embodiment,
the second ILM is a derivative of Formula (VI), or an unnatural
mimetic thereof. In a certain embodiment, the at least one
additional independently selected linker group comprises two
additional independently selected linker groups chemically linking
the ILM and the second ILM. In an embodiment, the at least one
additional linker group for an ILM of the Formula (VI), or an
unnatural mimetic thereof, chemically links groups selected from
R.sub.4 and R.sub.5. For example, an ILM of Formula (VI) and a
second ILM of Formula (VI), or an unnatural mimetic thereof, can be
linked as shown below:
##STR00004##
[0127] In certain embodiments, the ILM, the at least one additional
independently selected linker group L, and the second ILM has a
structure selected from the group consisting of:
##STR00005## ##STR00006##
[0128] which are derivatives of IAP antagonists described in WO
Pub. No. 2008/014236.
[0129] In any of the compounds described herein, the ILM can have
the structure of Formula (VIII), which is based on the IAP ligrands
described in Ndubaku, C., et al. Antagonism of c-IAP and XIAP
proteins is required for efficient induction of cell death by
small-molecule IAP antagonists, ACS Chem. Biol., 557-566, 4 (7)
(2009), or an unnatural mimetic thereof:
##STR00007##
[0130] wherein each of A1 and A2 of Formula (VIII) is independently
selected from optionally substituted monocyclic, fused rings, aryls
and hetoroaryls; and
[0131] R of Formula (VIII) is selected from H or Me.
[0132] In a particular embodiment, the linker group L is attached
to A1 of Formula (VIII). In another embodiment, the linker group L
is attached to A2 of Formula (VIII).
[0133] In a particular embodiment, the ILM is selected from the
group consisting of
##STR00008##
[0134] In any of the compounds described herein, the ILM can have
the structure of Formula (IX), which is derived from the chemotypes
cross-referenced in Mannhold, R., et al. IAP antagonists: promising
candidates for cancer therapy, Drug Discov. Today, 15 (5-6), 210-9
(2010), or an unnatural mimetic thereof:
##STR00009##
[0135] wherein R.sup.1 is selected from alkyl, cycloalkyl and
heterocycloalkyl and, most preferably, from isopropyl, tert-butyl,
cyclohexyl and tetrahydropyranyl, and R.sup.2 of Formula (IX) is
selected from --OPh or H.
[0136] In any of the compounds described herein, the ILM can have
the structure of Formula (X), which is derived from the chemotypes
cross-referenced in Mannhold, R., et al. IAP antagonists: promising
candidates for cancer therapy, Drug Discov. Today, 15 (5-6), 210-9
(2010), or an unnatural mimetic thereof:
##STR00010##
[0137] wherein:
[0138] R.sup.1 of Formula (X) is selected from H, --CH.sub.2OH,
--CH.sub.2CH.sub.2OH, --CH.sub.2NH.sub.2,
--CH.sub.2CH.sub.2NH.sub.2;
[0139] X of Formula (X) is selected from S or CH.sub.2;
[0140] R.sup.2 of Formula (X) is selected from:
##STR00011##
[0141] R.sup.3 and R.sup.4 of Formula (X) are independently
selected from H or Me
[0142] In any of the compounds described herein, the ILM can have
the structure of Formula (XI), which is derived from the chemotypes
cross-referenced in Mannhold, R., et al. IAP antagonists: promising
candidates for cancer therapy, Drug Discov. Today, 15 (5-6), 210-9
(2010), or an unnatural mimetic thereof:
##STR00012##
[0143] wherein R.sup.1 of Formula (XI) is selected from H or Me,
and R.sup.2 of Formula (XI) is selected from H or
##STR00013##
[0144] In any of the compounds described herein, the ILM can have
the structure of Formula (XII), which is derived from the
chemotypes cross-referenced in Mannhold, R., et al. IAP
antagonists: promising candidates for cancer therapy, Drug Discov.
Today, 15 (5-6), 210-9 (2010), or an unnatural mimetic thereof:
##STR00014##
wherein: R.sup.1 of Formula (XII) is selected from:
##STR00015##
and R.sup.2 of Formula (XII) is selected from:
##STR00016##
[0145] In any of the compounds described herein, the IAP E3
ubiquitin ligase binding moiety is selected from the group
consisting of:
##STR00017## ##STR00018## ##STR00019##
[0146] In any of the compounds described herein, the ILM can have
the structure of Formula (XIII), which is based on the IAP ligands
summarized in Flygare, J. A., et al. Small-molecule pan-IAP
antagonists: a patent review, Expert Opin. Ther. Pat., 20 (2),
251-67 (2010), or an unnatural mimetic thereof:
##STR00020##
wherein:
[0147] Z of Formula (XIII) is absent or O;
[0148] R.sup.1 of Formula (XIII) is selected from:
##STR00021##
[0149] R.sup.10 of
##STR00022##
is selected from H, alkyl, or aryl;
[0150] X is selected from CH2 and O; and
##STR00023##
is a nitrogen-containing heteroaryl.
[0151] In any of the compounds described herein, the ILM can have
the structure of Formula (XIV), which is based on the IAP ligands
summarized in Flygare, J. A., et al. Small-molecule pan-IAP
antagonists: a patent review, Expert Opin. Ther. Pat., 20 (2),
251-67 (2010), or an unnatural mimetic thereof:
##STR00024##
wherein:
[0152] Z of Formula (XIV) is absent or O;
[0153] R.sup.3 and R.sup.4 of Formula (XIV) are independently
selected from H or Me;
[0154] R.sup.1 of Formula (XIV) is selected from:
##STR00025##
[0155] R.sup.10 of
##STR00026##
is selected from H, alkyl, or aryl;
[0156] X of
##STR00027##
is selected from CH2 and O; and
##STR00028##
of or is a nitrogen-containing heteraryl.
[0157] In any of the compounds described herein, the ILM is
selected from the group consisting of:
##STR00029##
[0158] which are derivatives of ligands disclose in US Patent Pub.
No. 2008/0269140 and U.S. Pat. No. 7,244,851.
[0159] In any of the compounds described herein, the ILM can have
the structure of Formula (XV), which was a derivative of the IAP
ligand described in WO Pub. No. 2008/128171, or an unnatural
mimetic thereof:
##STR00030##
wherein:
[0160] Z of Formula (XV) is absent or O;
[0161] R.sup.1 of Formula (XV) is selected from:
##STR00031##
[0162] R.sup.10 of
##STR00032##
is selected from H, alkyl, or aryl;
[0163] X of
##STR00033##
is selected from CH2 and O; and
##STR00034##
is a nitrogen-containing heteraryl; and
[0164] R.sup.2 of Formula (XV) selected from H, alkyl, or acyl;
[0165] In a particular embodiment, the ILM has the following
structure:
##STR00035##
[0166] In any of the compounds described herein, the ILM can have
the structure of Formula (XVI), which is based on the IAP ligand
described in WO Pub. No. 2006/069063, or an unnatural mimetic
thereof:
##STR00036##
[0167] wherein: [0168] R.sup.2 of Formula (XVI) is selected from
alkyl, cycloalkyl and heterocycloalkyl; more preferably, from
isopropyl, tert-butyl, cyclohexyl and tetrahydropyranyl, most
preferably from cyclohexyl;
##STR00037##
[0168] of Formula (XVI) is a 5- or 6-membered nitrogen-containing
heteroaryl; more preferably, 5-membered nitrogen-containing
heteroaryl, and most preferably thiazole; and Ar of Formula (XVI)
is an aryl or a heteroaryl.
[0169] In any of the compounds described herein, the ILM can have
the structure of Formula (XVII), which is based on the IAP ligands
described in Cohen, F. et al., Antogonists of inhibitors of
apoptosis proteins based on thiazole amide isosteres, Bioorg. Med.
Chem. Lett., 20(7), 2229-33 (2010), or an unnatural mimetic
thereof:
##STR00038##
[0170] wherein:
[0171] R.sup.1 of Formula (XVII) is selected from te group halogen
(e.g. fluorine), cyano,
##STR00039##
[0172] X of Formula (XVII) is selected from the group O or CH2.
[0173] In any of the compounds described herein, the ILM can have
the structure of Formula (XVIII), which is based on the IAP ligands
described in Cohen, F. et al., Antogonists of inhibitors of
apoptosis proteins based on thiazole amide isosteres, Bioorg. Med.
Chem. Lett., 20(7), 2229-33 (2010), or an unnatural mimetic
thereof:
##STR00040##
[0174] wherein R of Formula (XVIII) is selected from alkyl, aryl,
heteroaryl, arylalkyl, heteroarylalkyl or halogen (in variable
substitution position).
[0175] In any of the compounds described herein, the ILM can have
the structure of Formula (XIX), which is based on the IAP ligands
described in Cohen, F. et al., Antogonists of inhibitors of
apoptosis proteins based on thiazole amide isosteres, Bioorg. Med.
Chem. Lett., 20(7), 2229-33 (2010), or an unnatural mimetic
thereof:
##STR00041##
[0176] wherein
##STR00042##
is a 6-member nitrogen heteroaryl.
[0177] In a certain embodiment, the ILM of the composition is
selected from the group consisting of:
##STR00043##
[0178] In certain embodiments, the ILM of the composition is
selected from the group consisting of:
##STR00044##
[0179] In any of the compounds described herein, the ILM can have
the structure of Formula (XX), which is based on the IAP ligands
described in WO Pub. No. 2007/101347, or an unnatural mimetic
thereof:
##STR00045##
[0180] wherein X of Formula (XX) is selected from CH.sub.2, O, NH,
or S.
[0181] In any of the compounds described herein, the ILM can have
the structure of Formula (XXI), which is based on the IAP ligands
described in U.S. Pat. Nos. 7,345,081 and 7,419,975, or an
unnatural mimetic thereof:
##STR00046##
wherein:
[0182] R.sup.2 of Formula (XXI) is selected from:
##STR00047##
[0183] R.sup.5 of Formula (XXI) is selected from:
##STR00048##
and
[0184] W of Formula (XXI) is selected from CH or N; and
[0185] R.sup.6 of
##STR00049##
and are independently a mono- or bicyclic fused aryl or
heteroaryl.
[0186] In certain embodiments, the ILM of the compound is selected
from the group consisting of:
##STR00050##
[0187] In certain embodiments, the ILM of the compound is selected
from the group consisting of:
##STR00051## ##STR00052##
which are described in WO Pub. No. 2009/060292, U.S. Pat. No.
7,517,906, WO Pub. No. 2008/134679, WO Pub. No. 2007/130626, and WO
Pub. No. 2008/128121.
[0188] In any of the compounds described herein, the ILM can have
the structure of Formula (XXII) or (XXIII), which are derived from
the IAP ligands described in WO Pub. No. 2015/006524 and Perez H L,
Discovery of potent heterodimeric antagonists of inhibitor of
apoptosis proteins (IAPs) with sustained antitumor activity. J.
Med. Chem. 58(3), 1556-62 (2015), or an unnatural mimetic
thereof:
##STR00053##
wherein: [0189] R.sup.1 of Formula (XII) or (XXII) is optionally
substituted alkyl, optionally substituted cycloalkyl, optionally
substituted cycloalkylalkyl, optionally substituted heterocyclyl,
optionally substituted arylalkyl or optionally substituted aryl;
[0190] R.sup.2 of Formula (XXII) or (XXIII) is optionally
substituted alkyl, optionally substituted cycloalkyl, optionally
substituted cycloalkylalkyl, optionally substituted heterocyclyl,
optionally substituted arylalkyl or optionally substituted aryl;
[0191] or alternatively, R.sup.1 and R.sup.2 of Formula (XXII) or
(XXIII) are independently optionally substituted thioalkyl wherein
the substituents attached to the S atom of the thioalkyl are
optionally substituted alkyl, optionally substituted branched
alkyl, optionally substituted heterocyclyl,
--(CH.sub.2).sub.vCOR.sup.20, --CH.sub.2CHR.sup.21COR.sup.22 or
--CH.sub.2R.sup.23; [0192] wherein: [0193] v is an integer from
1-3; [0194] R.sup.20 and R.sup.22 of --(CH.sub.2).sub.vCOR.sup.20
and --CH.sub.2R.sup.23 are independently selected from OH,
NR.sup.24R.sup.25 or OR.sup.26; [0195] R.sup.21 of
--CH.sub.2CHR.sup.21COR.sup.2 is selected from the group
NR.sup.24R.sup.25; [0196] R.sup.23 of --CH.sub.2R.sup.23 is
selected from optionally substituted aryl or optionally substituted
heterocyclyl, where the optional substituents include alkyl and
halogen; [0197] R.sup.24 of NR.sup.24R.sup.25 is selected from
hydrogen or optionally substituted alkyl; [0198] R.sup.25 of
NR.sup.24R.sup.25 is selected from hydrogen, optionally substituted
alkyl, optionally substituted branched alkyl, optionally
substituted arylalkyl, optionally substituted heterocyclyl,
--CH.sub.2(OCH.sub.2CH.sub.2O).sub.mCH.sub.3, or a polyamine chain,
such as spermine or spermidine; [0199] R.sup.26 of OR.sup.26 is
selected from optionally substituted alkyl, wherein the optional
substituents are OH, halogen or NH.sub.2; and [0200] m is an
integer from 1-8; [0201] R.sup.3 and R.sup.4 of Formula (XXII) or
(XXIII) are independently selected from optionally substituted
alkyl, optionally substituted cycloalkyl, optionally substituted
aryl, optionally substituted arylalkyl, optionally substituted
arylalkoxy, optionally substituted heteroaryl, optionally
substituted heterocyclyl, optionally substituted heteroarylalkyl or
optionally substituted heterocycloalkyl, wherein the substituents
are alkyl, halogen or OH; [0202] R.sup.5, R.sup.6, R.sup.7 and
R.sup.8 of Formula (XXII) or (XXIII) are independently selected
from hydrogen, optionally substituted alkyl or optionally
substituted cycloalkyl; and [0203] X is selected from a bond or a
chemical linker group, and/or a pharmaceutically acceptable salt,
tautomer or stereoisomer thereof.
[0204] In certain embodiments, X is a bond or is selected from the
group consisting of:
##STR00054## ##STR00055##
wherein "*" is the point of attachment of a PTM, L or ULM, e.g., an
ILM.
[0205] In any of the compounds described herein, the ILM can have
the structure of Formula (XXIV) or (XXVI), which are derived from
the IAP ligands described in WO Pub. No. 2015/006524 and Perez H L,
Discovery of potent heterodimeric antagonists of inhibitor of
apoptosis proteins (IAPs) with sustained antitumor activity. J.
Med. Chem. 58(3), 1556-62 (2015), or an unnatural mimetic thereof,
and the chemical linker to linker group L as shown:
##STR00056##
wherein: [0206] R.sup.1 of Formula (XXIV), (XXV) or (XXVI) is
selected from optionally substituted alkyl, optionally substituted
cycloalkyl, optionally substituted cycloalkylalkyl, optionally
substituted heterocyclyl, optionally substituted arylalkyl or
optionally substituted aryl; [0207] R.sup.2 of Formula (XXIV),
(XXV) or (XXVI) is selected from optionally substituted alkyl,
optionally substituted cycloalkyl, optionally substituted
cycloalkylalkyl, optionally substituted heterocyclyl, optionally
substituted arylalkyl or optionally substituted aryl; [0208] or
alternatively, [0209] R.sup.1 and R.sup.2 of Formula (XXIV), (XXV)
or (XXVI) are independently selected from optionally substituted
thioalkyl wherein the substituents attached to the S atom of the
thioalkyl are optionally substituted alkyl, optionally substituted
branched alkyl, optionally substituted heterocyclyl,
--(CH.sub.2).sub.vCOR.sup.20, --CH.sub.2CHR.sup.21COR.sup.22 or
--CH.sub.2R.sup.23, [0210] wherein: [0211] v is an integer from
1-3; [0212] R.sup.20 and R.sup.22 of --(CH.sub.2).sub.vCOR.sup.20
and --CH.sub.2R.sup.23 are independently selected from OH,
NR.sup.24R.sup.25 or OR.sup.26; [0213] R.sup.21 of
--CH.sub.2CHR.sup.21COR.sup.2 is selected from NR.sup.24R.sup.25;
[0214] R.sup.23 of --CH.sub.2R.sup.23 is selected from optionally
substituted aryl or optionally substituted heterocyclyl, wherein
the optional substituents include alkyl and halogen; [0215]
R.sup.24 of NR.sup.24R.sup.25 is selected from hydrogen or
optionally substituted alkyl; [0216] R.sup.25 of NR.sup.24R.sup.25
is selected from hydrogen, optionally substituted alkyl, optionally
substituted branched alkyl, optionally substituted arylalkyl,
optionally substituted heterocyclyl,
--CH.sub.2(OCH.sub.2CH.sub.2O).sub.mCH.sub.3, or a polyamine chain,
such as spermine or spermidine; [0217] R.sup.26 of OR.sup.26 is
selected from optionally substituted alkyl, wherein the optional
substituents are OH, halogen or NH.sub.2; and [0218] m is an
integer from 1-8; [0219] R.sup.3 and R.sup.4 of Formula (XXIV),
(XXV) or (XXVI) are independently optionally substituted alkyl,
optionally substituted cycloalkyl, optionally substituted aryl,
optionally substituted arylalkyl, optionally substituted
arylalkoxy, optionally substituted heteroaryl, optionally
substituted heterocyclyl, optionally substituted heteroarylalkyl or
optionally substituted heterocycloalkyl, wherein the substituents
are alkyl, halogen or OH; [0220] R.sup.5, R.sup.6, R.sup.7 and
R.sup.8 of Formula (XXIV), (XXV) or (XXVI) are independently
hydrogen, optionally substituted alkyl or optionally substituted
cycloalkyl; and/or a pharmaceutically acceptable salt, tautomer or
stereoisomer thereof.
[0221] In a particular embodiment, the ILM according to Formulas
(XXII) through (XXVI):
R.sup.7 and R.sup.8 are selected from the H or Me; R.sup.5 and
R.sup.6 are selected from the group comprising:
##STR00057##
R.sup.3 and R.sup.4 are selected from the group comprising:
##STR00058##
[0222] In any of the compounds described herein, the ILM can have
the structure of Formula (XXVII) or (XXVII), which are derived from
the IAP ligands described in WO Pub. No. 2014/055461 and Kim, K S,
Discovery of tetrahydroisoquinoline-based bivalent heterodimeric
IAP antagonists. Bioorg. Med. Chem. Lett. 24(21), 5022-9 (2014), or
an unnatural mimetic thereof:
##STR00059##
wherein: [0223] R.sup.35 is 1-2 substituents selected from alkyl,
halogen, alkoxy, cyano and haloalkoxy; [0224] R.sup.1 of Formula
(XXVII) and (XXVIII) is selected from H or an optionally
substituted alkyl, optionally substituted cycloalkyl, optionally
substituted cycloalkylalkyl, optionally substituted heterocyclyl,
optionally substituted arylalkyl or optionally substituted aryl;
[0225] R.sup.2 of Formula (XXVII) and (XXVIII) is selected from H
or an optionally substituted alkyl, optionally substituted
cycloalkyl, optionally substituted cycloalkylalkyl, optionally
substituted heterocyclyl, optionally substituted arylalkyl or
optionally substituted aryl; [0226] or alternatively, [0227]
R.sup.1 and R.sup.2 of Formula (XXVII) and (XXVIII) are
independently selected from an optionally substituted thioalkyl
--CR.sup.60R.sup.61SR.sup.70, wherein R.sup.60 and R.sup.61 are
selected from H or methyl, and R.sup.70 is selected from an
optionally substituted alkyl, optionally substituted branched
alkyl, optionally substituted heterocyclyl,
--(CH.sub.2).sub.vCOR.sup.20, --CH.sub.2CHR.sup.21COR.sup.22 or
--CH.sub.2R.sup.23, [0228] wherein: [0229] v is an integer from
1-3; [0230] R.sup.20 and R.sup.22 of --(CH.sub.2).sub.vCOR.sup.20
and --CH.sub.2CHR.sup.21COR.sup.22 are independently selected from
OH, NR.sup.24R.sup.25 or OR.sup.26; [0231] R.sup.21 of
--CH.sub.2CHR.sup.21COR.sup.22 is selected from NR.sup.24R.sup.25;
[0232] R.sup.23 of --CH.sub.2R.sup.23 is selected from an
optionally substituted aryl or optionally substituted heterocyclyl,
where the optional substituents include alkyl and halogen; [0233]
R.sup.24 of NR.sup.24R.sup.25 is selected from hydrogen or
optionally substituted alkyl; [0234] R.sup.25 of NR.sup.24R.sup.25
is selected from hydrogen, optionally substituted alkyl, optionally
substituted branched alkyl, optionally substituted arylalkyl,
optionally substituted heterocyclyl,
--CH.sub.2CH.sub.2(OCH.sub.2CH.sub.2).sub.mCH.sub.3, or a polyamine
chain
--[CH.sub.2CH.sub.2(CH.sub.2).sub..delta.NH].sub..psi.CH.sub.2CH.sub.2(CH-
.sub.2).omega.NH.sub.2, such as spermine or spermidine; [0235]
wherein .delta.=0-2, .psi.=1-3, .omega.=0-2; [0236] R.sup.26 of
OR.sup.26 is an optionally substituted alkyl, wherein the optional
substituents are OH, halogen or NH.sub.2; and [0237] m is an
integer from 1-8, [0238] R.sup.3 and R.sup.4 of Formula (XXVII) and
(XXVIII) are independently selected from an optionally substituted
alkyl, optionally substituted cycloalkyl, optionally substituted
aryl, optionally substituted arylalkyl, optionally substituted
arylalkoxy, optionally substituted heteroaryl, optionally
substituted heterocyclyl, optionally substituted heteroarylalkyl or
optionally substituted heterocycloalkyl, wherein the substituents
are alkyl, halogen or OH; [0239] R.sup.5, R.sup.6, R.sup.7 and
R.sup.8 of Formula (XXVII) and (XXVIII) are independently selected
from hydrogen, optionally substituted alkyl or optionally
substituted cycloalkyl; [0240] R.sup.31 of Formulas (XXVII) and
(XXVIII) is selected from alkyl, aryl, arylalkyl, heteroaryl or
heteroarylalkyl optionally further substituted, preferably selected
form the group consisting of:
[0240] ##STR00060## [0241] X of Formulas (XXVII) and (XXVIII) is
selected from --(CR.sup.81R.sup.82).sub.m--, optionally substituted
heteroaryl or heterocyclyl,
[0241] ##STR00061## [0242] Z of Formulas (XXVII) is selected from
C.dbd.O, --O--, --NR, --CONH--, --NHCO--, or may be absent; [0243]
R.sup.81 and R.sup.82 of --(CR.sup.81R.sup.82).sub.m-- are
independently selected from hydrogen, halogen, alkyl or cycloalkyl,
or R.sup.81 and R.sup.82 can be taken together to form a
carbocyclic ring; [0244] R.sup.10 and R.sup.11 of
[0244] ##STR00062## are independently selected from hydrogen,
halogen or alkyl; [0245] R.sup.12, R.sup.13, R.sup.14, R.sup.15 and
R.sup.16 of
[0245] ##STR00063## and are independently selected from hydrogen,
halogen or optionally substituted alkyl or OR.sup.17; [0246]
R.sup.17 is selected from hydrogen, optionally substituted alkyl or
optionally substituted cycloalkyl; [0247] m and n of
--(CR.sup.21R.sup.22).sub.m-- and
[0247] ##STR00064## are independently 0, 1, 2, 3, or 4; [0248] o
and p of
[0248] ##STR00065## are independently 0, 1, 2 or 3; [0249] q and t
of
[0249] ##STR00066## are independently 0, 1, 2, 3, or 4; [0250] r
of
##STR00067##
[0250] is 0 or 1; [0251] and/or a pharmaceutically acceptable salt,
tautomer or stereoisomer thereof.
[0252] In any of the compounds described herein, the ILM can have
the structure of Formula (XXIX), (XXX), (XXXI), or (XXXII), which
are derived from the IAP ligands described in WO Pub. No.
2014/055461 and Kim, K S, Discovery of tetrahydroisoquinoline-based
bivalent heterodimeric IAP antagonists. Bioorg. Med. Chem. Lett.
24(21), 5022-9 (2014), or an unnatural mimetic thereof, and the
chemical linker to linker group L as shown:
##STR00068##
wherein: [0253] R.sup.2 of Formula (XXIX) through (XXXII) is
selected from H, an optionally substituted alkyl, optionally
substituted cycloalkyl, optionally substituted cycloalkylalkyl,
optionally substituted heterocyclyl, optionally substituted
arylalkyl or optionally substituted aryl; [0254] or alternatively;
[0255] R.sup.1 and R.sup.2 of Formula (XXVII) and (XXVIII) are
independently selected from H, an optionally substituted thioalkyl
--CR.sup.60R.sup.61SR.sup.70 wherein R.sup.60 and R.sup.61 are
selected from H or methyl, and R.sup.70 is an optionally
substituted alkyl, optionally substituted branched alkyl,
optionally substituted heterocyclyl, --(CH.sub.2).sub.vCOR.sup.20,
--CH.sub.2CHR.sup.21COR.sup.22 or --CH.sub.2R.sup.23. [0256]
wherein: [0257] v is an integer from 1-3; [0258] R.sup.20 and
R.sup.22 of --(CH.sub.2).sub.vCOR.sup.20 and
--CH.sub.2CHR.sup.21COR.sup.22 are independently selected from OH,
NR.sup.24R.sup.25 or OR.sup.26; [0259] R.sup.21 of
--CH.sub.2CHR.sup.21COR.sup.22 is selected from NR.sup.24R.sup.25;
[0260] R.sup.23 of --CH.sub.2R.sup.23 is selected from an
optionally substituted aryl or optionally substituted heterocyclyl,
where the optional substituents include alkyl and halogen; [0261]
R.sup.24 of NR.sup.24R.sup.25 is selected from hydrogen or
optionally substituted alkyl; [0262] R.sup.25 of NR.sup.24R.sup.25
is selected from hydrogen, optionally substituted alkyl, optionally
substituted branched alkyl, optionally substituted arylalkyl,
optionally substituted heterocyclyl,
--CH.sub.2CH.sub.2(OCH.sub.2CH.sub.2).sub.mCH.sub.3, or a polyamine
chain
--[CH.sub.2CH.sub.2(CH.sub.2).sub..delta.NH].sub..psi.CH.sub.2CH.sub.2(CH-
.sub.2).omega..sub.rNH.sub.2, such as spermine or spermidine,
[0263] wherein .delta.=0-2, .psi.=1-3, .omega.=0-2; [0264] R.sup.26
of OR.sup.26 is an optionally substituted alkyl, wherein the
optional substituents are OH, halogen or NH.sub.2; [0265] m is an
integer from 1-8; [0266] R.sup.6 and R.sup.8 of Formula (XXIX)
through (XXXII) are independently selected from hydrogen,
optionally substituted alkyl or optionally substituted cycloalkyl;
and [0267] R.sup.31 of Formulas (XXIX) through (XXXII) is selected
from alkyl, aryl, arylalkyl, heteroaryl or heteroarylalkyl
optionally further substituted, preferably selected form the group
consisting of:
##STR00069##
[0268] In certain embodiments, the ILM of the compound is:
##STR00070##
[0269] In any of the compounds described herein, the ILM can have
the structure of Formula (XXXIII), which are derived from the IAP
ligands described in WO Pub. No. 2014/074658 and WO Pub. No.
2013/071035, or an unnatural mimetic thereof:
##STR00071##
wherein: [0270] R.sup.2 of Formula (XXXIII) is selected from H, an
optionally substituted alkyl, optionally substituted cycloalkyl,
optionally substituted cycloalkylalkyl, optionally substituted
heterocyclyl, optionally substituted arylalkyl or optionally
substituted aryl; [0271] R.sup.6 and R.sup.8 of Formula (XXXIII)
are independently selected from hydrogen, optionally substituted
alkyl or optionally substituted cycloalkyl; [0272] R.sup.32 of
Formula (XXXIII) is selected from (C.sub.1-C.sub.4
alkylene)-R.sup.33 wherein R.sup.33 is selected from hydrogen,
aryl, heteroaryl or cycloalkyl optionally further substituted;
[0273] X of Formula (XXXIII) is selected from:
[0273] ##STR00072## [0274] Z and Z' of Formula (XXXIII) are
independently selected from:
##STR00073##
[0274] wherein each
##STR00074##
represents a point of attachment to the compound, and Z and Z'
cannot both be
##STR00075##
in any given compound; [0275] Y of Formula (XXXIII) is selected
from:
##STR00076## ##STR00077##
[0275] wherein Z and Z' of Formula (XXXIII) are the same and Z
is
##STR00078##
wherein each
##STR00079##
represents a point of attachment to the compound, X is selected
from:
##STR00080## ##STR00081## ##STR00082##
and [0276] Y of Formula (XXXIII) is independently selected
from:
[0276] ##STR00083## ##STR00084## ##STR00085## [0277] wherein:
[0277] ##STR00086## represents a point of attachment to a --C.dbd.O
portion of the compound;
##STR00087## represents a point of attachment to a --NH portion of
the compound;
##STR00088## represents a first point of attachment to Z;
##STR00089## represents a second point of attachment to Z; [0278] m
is an integer from 0-3; [0279] n is an integer from 1-3; [0280] p
is an integer from 0-4; and [0281] A is --C(O)R.sup.3; [0282]
R.sup.3 is selected from --C(O)R.sup.3 is OH, NHCN,
NHSO.sub.2R.sup.10, NHOR.sup.11 or N(R.sup.12)(R.sup.13); [0283]
R.sup.10 and F.sup.11 of NHSO.sub.2R.sup.10 and NHOR.sup.11 are
independently selected from hydrogen, optionally substituted
--C.sub.1-C.sub.4 alkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl
or heterocycloalkyl; [0284] R.sup.12 and R.sup.13 of
N(R.sup.12)(R.sup.13) are independently selected from hydrogen,
--C.sub.1-C.sub.4 alkyl, --(C.sub.1-C.sub.4)
alkylene)-NH--(C.sub.1-C.sub.4 alkyl), and --(C.sub.1-C.sub.4
alkylene)-O--(C.sub.1-C.sub.4 hydroxyalkyl), or R.sup.12 and
R.sup.13 taken together with the nitrogen atom to which they are
commonly bound to form a saturated heterocyclyl optionally
comprising one additional heteroatom selected from N, O and S, and
wherein the saturated heterocycle is optionally substituted with
methyl.
[0285] In any of the compounds described herein, the ILM can have
the structure of Formula (XXXIV) or (XXXV), which are derived from
the IAP ligands described in WO Pub. No. 2014/047024, or an
unnatural mimetic thereof:
##STR00090##
wherein: [0286] X of Formula (XXXIV) or (XXXV) is absent or a group
selected from --(CR.sup.10R.sup.11).sub.m--, optionally substituted
heteroaryl or optionally substituted heterocyclyl,
[0286] ##STR00091## [0287] Y and Z of Formula (XXXIV) or (XXXV) are
independently selected from C.dbd.O, --O--, --NR.sup.9--, --CONH--,
--NHCO-- or may be absent; [0288] R.sup.1 and R.sup.2 of Formula
(XXXIV) or (XXXV) are independently selected from an optionally
substituted alkyl, optionally substituted cycloalkyl, optionally
substituted cycloalkylalkyl, optionally substituted arylalkyl,
optionally substituted aryl, or [0289] R.sup.1 and R.sup.2 of
Formula (XXXIV) or (XXXV) are independently selected from
optionally substituted thioalkyl wherein the substituents attached
to the S atom of the thioalkyl are optionally substituted alkyl,
optionally substituted branched alkyl, optionally substituted
heterocyclyl, --(CH.sub.2).sub.vCOR.sup.20,
--CH.sub.2CHR.sup.21COR.sup.22 or --CH.sub.2R.sup.23; wherein v is
an integer from 1-3; [0290] R.sup.20 and R.sup.22 of
--(CH.sub.2).sub.vCOR.sup.20 and --CH.sub.2CHR.sup.21COR.sup.22 are
independently selected from OH, NR.sup.24R.sup.25 or OR.sup.26;
[0291] R.sup.21 of --CH.sub.2CHR.sup.21COR.sup.22 is selected from
NR.sup.24R.sup.25; [0292] R.sup.23 of --CH.sub.2R.sup.23 are
selected from an optionally substituted aryl or optionally
substituted heterocyclyl, where the optional substituents include
alkyl and halogen; [0293] R.sup.24 of NR.sup.24R.sup.25 is selected
from hydrogen or optionally substituted alkyl; [0294] R.sup.25 of
NR.sup.24R.sup.25 is selected from hydrogen, optionally substituted
alkyl, optionally substituted branched alkyl, optionally
substituted arylalkyl, optionally substituted heterocyclyl,
--CH.sub.2(OCH.sub.2CH.sup.20)mCH3, or a polyamine chain; [0295]
R.sup.26 is an optionally substituted alkyl, wherein the optional
substituents are OH, halogen or NH.sub.2; [0296] m of
--(CR.sup.10R.sup.11).sub.m is an integer from 1-8; [0297] R.sup.3
and R.sup.4 of Formula (XXXIV) or (XXXV) are independently selected
from optionally substituted alkyl, optionally substituted
cycloalkyl, optionally substituted aryl, optionally substituted
arylalkyl, optionally substituted arylalkoxy, optionally
substituted heteroaryl, optionally substituted heterocyclyl,
optionally substituted heteroarylalkyl or optionally substituted
heterocycloalkyl, wherein the substituents are alkyl, halogen or
OH; [0298] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 of Formula (XXXIV)
or (XXXV) are independently selected from hydrogen, optionally
substituted alkyl or optionally substituted cycloalkyl; [0299]
R.sup.10 and R.sup.11 of --(CR.sup.10R.sup.11).sub.m are
independently selected from hydrogen, halogen or optionally
substituted alkyl; [0300] R.sup.12 and R.sup.13 of
##STR00092##
[0300] are independently selected from hydrogen, halogen or
optionally substituted alkyl, or R.sup.12 and R.sup.13 can be taken
together to form a carbocyclic ring; [0301] R.sup.14, R.sup.15,
R.sup.16, R.sup.17 and R.sup.18 of
##STR00093##
[0301] and are independently selected from hydrogen, halogen,
optionally substituted alkyl or OR.sup.19; [0302] R.sup.19 of
OR.sup.19 is selected from hydrogen, optionally substituted alkyl
or optionally substituted cycloalkyl; [0303] m and n of
--(CR.sup.10R.sup.11).sub.m are independently 0, 1, 2, 3, or 4;
[0304] o and p of --(CR.sup.10R.sup.11).sub.m are independently 0,
1, 2 or 3; [0305] q of --(CR.sup.10R.sup.11).sub.m is 0, 1, 2, 3,
or 4; r is 0 or 1; [0306] t of --(CR.sup.10R.sup.11).sub.m is 1, 2,
or 3; and/or a pharmaceutically acceptable salt, tautomer or
stereoisomer thereof.
[0307] In any of the compounds described herein, the ILM can have
the structure of Formula (XXXVI), which are derived from the IAP
ligands described in WO Pub. No. 2014/025759, or an unnatural
mimetic thereof:
##STR00094##
where: [0308] A of Formula (XXXVI) is selected from:
##STR00095##
[0308] where the dotted line represents an optional double bond;
[0309] X of Formula (XXXVI) is selected from:
--(CR.sup.21R.sup.22).sub.m--,
[0309] ##STR00096## [0310] Y and Z of Formula (XXXVI) are
independently selected from -0-, --NR.sup.6-- or are absent; [0311]
V of Formula (XXXVI) is selected from --N-- or --CH--; [0312] W of
Formula (XXXVI) is selected from --CH-- or --N--; [0313] R.sup.1 of
Formula (XXXVI) is selected from an optionally substituted alkyl,
optionally substituted cycloalkyl, optionally substituted
cycloalkylalkyl, optionally substituted arylalkyl or optionally
substituted aryl; [0314] R.sup.3 and R.sup.4 of Formula (XXXVI) are
independently selected from optionally substituted alkyl,
optionally substituted cycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, optionally substituted
heterocyclyl, optionally substituted arylalkyl, optionally
substituted heteroarylalkyl or optionally substituted
heterocycloalkyl; [0315] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 of
Formula (XXIV), (XXV) or (XXVI) are independently selected from
hydrogen, optionally substituted alkyl or optionally substituted
cycloalkyl, or preferably methyl; [0316] R.sup.9 and R.sup.10
of
##STR00097##
[0316] are independently selected from hydrogen, halogen or
optionally substituted alkyl, or R.sup.9 and R.sup.10 can be taken
together to form a ring; [0317] R.sup.11, R.sup.12, R.sup.13 and
R.sup.14 of
##STR00098##
[0317] are independently selected from hydrogen, halogen,
optionally substituted alkyl or OR.sup.15; [0318] R.sup.15 of
OR.sup.15 is selected from hydrogen, optionally substituted alkyl
or optionally substituted cycloalkyl; [0319] m and n of
--(CR.sup.21R.sup.22).sub.m-- and
##STR00099##
[0319] are independently selected from 0, 1, 2, 3, or 4; [0320] o
and p of
##STR00100##
[0320] and are independently selected from 0, 1, 2 or 3; [0321] q
of
##STR00101##
[0321] is selected from 0, 1, 2, 3, or 4; [0322] r of
##STR00102##
[0322] is selected from 0 or 1, and/or or a pharmaceutically
acceptable salt, tautomer or stereoisomer thereof.
[0323] In any of the compounds described herein, the ILM can have
the structure of Formula (XXXVII) or (XXXVIII), which are derived
from the IAP ligands described in WO Pub. No. 2014/011712, or an
unnatural mimetic thereof:
##STR00103##
wherein: [0324] X of Formulas (XXXVII) and (XXXVIII) is
--(CR.sup.16R.sup.17).sub.m--,
##STR00104##
[0324] or absent; [0325] Y and Z of Formula (XXXVII) and (XXXVIII)
are independently selected from -0-, C=0, NR.sup.6 or are absent;
[0326] R.sup.1 and R.sup.2 of Formula (XXXVII) and (XXXVIII) are
selected from optionally substituted alkyl, optionally substituted
cycloalkyl, optionally substituted alkylaryl or optionally
substituted aryl; [0327] R.sup.3 and R.sup.4 of Formula (XXXVII)
and (XXXVIII) are independently selected from optionally
substituted alkyl, optionally substituted cycloalkyl, optionally
substituted cycloalkylalkyl, optionally substituted arylalkyl or
optionally substituted aryl; [0328] R.sup.5 and R.sup.6 of Formula
(XXXVII) and (XXXVIII) are independently selected from optionally
substituted alkyl or optionally substituted cycloalkyl; [0329]
R.sup.7 and R.sup.8 of Formula (XXXVII) and (XXXVIII) are
independently selected from hydrogen, optionally substituted alkyl
or optionally substituted cycloalkyl, or preferrably methyl; [0330]
R.sup.9 and R.sup.10 of
##STR00105##
[0330] are independently selected from hydrogen, optionally
substituted alkyl, or R.sup.9 and R.sup.10 may be taken together to
form a ring; [0331] R.sup.11 to R.sup.14 of
##STR00106##
[0331] are independently selected from hydrogen, halogen,
optionally substituted alkyl or OR.sup.15; [0332] R.sup.15 of
OR.sup.15 is selected from hydrogen, optionally substituted alkyl
or optionally substituted cycloalkyl; [0333] R.sup.16 and R.sup.17
of --(CR.sup.16R.sup.17).sub.m-- are independently selected from
hydrogen, halogen or optionally substituted alkyl; [0334] R.sup.50
and R.sup.51 of Formula (XXXVII) and (XXXVIII) are independently
selected from optionally substituted alkyl, or R.sup.50 and
R.sup.51 are taken together to form a ring; [0335] m and n of
--(CR.sup.16R.sup.17).sub.m-- and
##STR00107##
[0335] are independently an integer from 0-4; [0336] o and p of
##STR00108##
[0336] are independently an integer from 0-3; [0337] q of
##STR00109##
[0337] is an integer from 0-4; and [0338] r of
##STR00110##
[0338] is an integer from 0-1; [0339] or a pharmaceutically
acceptable salt, tautomer or stereoisomer thereof.
[0340] In an embodiment, R.sup.1 and R.sup.2 of the ILM of Formula
(XXXVII) or (XXXVIII) are t-butyl and R.sup.3 and R.sup.4 of the
ILM of Formula (XXXVII) or (XXXVIII) are tetrahydronaphtalene.
[0341] In any of the compounds described herein, the ILM can have
the structure of Formula (XXXIX) or (XL), which are derived from
the IAP ligands described in WO Pub. No. 2013/071039, or an
unnatural mimetic thereof:
##STR00111##
wherein: [0342] R.sup.43 and R.sup.44 of Formulas (XXXIX) and (XL)
are independently selected from hydrogen, alkyl, aryl, arylalkyl,
heteroaryl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl further
optionally substituted, and [0343] R.sup.6 and R.sup.8 of Formula
(XXXIX) and (XL) are independently selected from hydrogen,
optionally substituted alkyl or optionally substituted cycloalkyl.
[0344] each X of Formulas (XXXIX) and (XL) is independently
selected from:
[0344] ##STR00112## ##STR00113## [0345] each Z of Formulas (XXXIX)
and (XL) is selected from
##STR00114##
[0345] wherein each
##STR00115##
represents a point of attachment to the compound; and [0346] each Y
is selected from:
##STR00116## ##STR00117## ##STR00118## ##STR00119##
[0346] wherein:
##STR00120##
represents a point of attachment to a --C.dbd.O portion of the
compound;
##STR00121##
represents a point of attachment to an amino portion of the
compound;
##STR00122## [0347] represents a first point of attachment to
Z;
[0347] ##STR00123## [0348] represents a second point of attachment
to Z; and [0349] A is selected from --C(O)R.sup.3 or
##STR00124##
[0349] or a tautomeric form of any of the foregoing, wherein:
[0350] R.sup.3 of --C(O)R.sup.3 is selected from OH, NHCN,
NHS0.sub.2R.sup.10, NHOR.sup.11 or N(R.sup.12)(R.sup.13); [0351]
R.sup.10 and R.sup.11 of NHS0.sub.2R.sup.10 and NHOR.sup.11 are
independently selected from --C.sub.1-C.sub.4 alkyl, cycloalkyl,
aryl, heteroaryl, or heterocycloalkyl, any of which are optionally
substituted, and hydrogen; [0352] each of R.sup.12 and R.sup.13 of
N(R.sup.12)(R.sup.13) are independently selected from hydrogen,
--C.sub.1-C.sub.4 alkyl, --(C.sub.1-C.sub.4
alkylene)-NH--(C.sub.1-C.sub.4 alkyl), benzyl, --(C.sub.1-C.sub.4
alkylene)-C(O)OH, [0353] --(C.sub.1-C.sub.4 alkylene)-C(O)CH3,
--CH(benzyl)-COOH, --C.sub.1-C.sub.4 alkoxy, and [0354]
--(C.sub.1-C.sub.4 alkylene)-O--(C.sub.1-C.sub.4 hydroxyalkyl); or
R.sup.12 and R.sup.13 of N(R.sup.12)(R.sup.13) are taken together
with the nitrogen atom to which they are commonly bound to form a
saturated heterocyclyl optionally comprising one additional
heteroatom selected from N, O and S, and wherein the saturated
heterocycle is optionally substituted with methyl.
[0355] In any of the compounds described herein, the ILM can have
the structure of Formula (XLI), which are derived from the IAP
ligands described in WO Pub. No. 2013/071039, or an unnatural
mimetic thereof:
##STR00125##
wherein: [0356] W.sup.1 of Formula (XLI) is selected from O, S,
N--R.sup.A, or C(R.sup.8a)(R.sup.8b); [0357] W.sup.2 of Formula
(XLI) is selected from O, S, N--R.sup.A, or C(R.sup.8c)(R.sup.8d);
provided that W.sup.1 and W.sup.2 are not both O, or both S; [0358]
R.sup.1 of Formula (XLI) is selected from H, C.sub.1-C.sub.6alkyl,
C.sub.3-C.sub.6cycloalkyl, --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.3-C.sub.6cycloalkyl), substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl);
[0359] when X.sup.1 is selected from O, N--R.sup.A, S, S(O), or
S(O).sub.2, then X.sup.2 is C(R.sup.2aR.sup.2b); [0360] or: [0361]
X.sup.1 of Formula (XLI) is selected from CR.sup.2cR.sup.2d and
X.sup.2 is CR.sup.2aR.sup.2b, and R.sup.2c and R.sup.2a together
form a bond; [0362] or: [0363] X.sup.1 and X.sup.2 of Formula (XLI)
are independently selected from C and N, and are members of a fused
substituted or unsubstituted saturated or partially saturated 3-10
membered cycloalkyl ring, a fused substituted or unsubstituted
saturated or partially saturated 3-10 membered heterocycloalkyl
ring, a fused substituted or unsubstituted 5-10 membered aryl ring,
or a fused substituted or unsubstituted 5-10 membered heteroaryl
ring; [0364] or: [0365] X.sup.1 of Formula (XLI) is selected from
CH.sub.2 and X.sup.2 is C.dbd.0, C.dbd.C(R.sup.C).sub.2, or
C.dbd.NR.sup.C; where each R.sup.c is independently selected from
H, --CN, --OH, alkoxy, substituted or unsubstituted
C.sub.1-C.sub.6alkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl);
[0366] R.sup.A of N--R.sup.A is selected from H,
C.sub.1-C.sub.6alkyl, --C(.dbd.O)C.sub.1-C.sub.2alkyl, substituted
or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
[0367] R.sup.2a, R.sup.2b, R.sup.2c, R.sup.2d of CR.sup.2cR.sup.2d
and CR.sup.2aR.sup.2b are independently selected from H,
substituted or unsubstituted C1-C6alkyl, substituted or
unsubstituted C.sub.1-C.sub.6heteroalkyl, substituted or
unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl)
and --C(.dbd.O)R.sup.B; [0368] R.sup.B of --C(.dbd.O)R.sup.B is
selected from substituted or unsubstituted C.sub.1-C.sub.6alkyl,
substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted
or unsubstituted C.sub.2-C.sub.5heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl),
or --NR.sup.DR.sup.E; [0369] R.sup.D and R.sup.E of NR.sup.DR.sup.E
are independently selected from H, substituted or unsubstituted
C.sub.1-C.sub.6alkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, --C.sub.1-C.sub.6alkyl-
(substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted heteroaryl);
[0370] m of Formula (XLI) is selected from 0, 1 or 2; [0371]
--U--of Formula (XLI) is selected from --NHC(.dbd.O)--,
--C(.dbd.O)NH--, --NHS(.dbd.O).sub.2--, --S(.dbd.O).sub.2NH--,
--NHC(.dbd.O)NH--, --NH(C.dbd.O)O--, --O(C.dbd.O)NH--, or
--NHS(.dbd.O).sub.2NH--; [0372] R.sup.3 of Formula (XLI) is
selected from C.sub.1-C.sub.3alkyl, or C.sub.1-C.sub.3fluoroalkyl;
[0373] R.sup.4 of Formula (XLI) is selected from --NHR.sup.5,
--N(R.sup.5)2, --N+(R.sup.5)3 or --OR.sup.5; [0374] each R.sup.5 of
--NHR.sup.5, --N(R.sup.5)2, --N+(R.sup.5)3 and --OR.sup.5 is
independently selected from H, C.sub.1-C.sub.3alkyl,
C.sub.1-C.sub.3haloalkyl, C.sub.1-C.sub.3heteroalkyl and
--C.sub.1-C.sub.3alkyl-(C.sub.3-C.sub.5cycloalkyl); [0375] or:
[0376] R.sup.3 and R.sup.5 of Formula (XLI) together with the atoms
to which they are attached form a substituted or unsubstituted 5-7
membered ring; [0377] or: [0378] R.sup.3 of Formula (XLI) is bonded
to a nitrogen atom of U to form a substituted or unsubstituted 5-7
membered ring; [0379] R.sup.6 of Formula (XLI) is selected from
--NHC(.dbd.O)R.sup.7, --C(.dbd.O)NHR.sup.7, --NHS(.dbd.O)2R.sup.7,
--S(.dbd.O).sub.2NHR.sup.7; --NHC(.dbd.O)NHR.sup.7,
--NHS(.dbd.O).sub.2NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)R.sup.7,
--(C.sub.1-C.sub.3alkyl)-C(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O)2R.sup.7,
--(C.sub.1-C.sub.3alkyl)-S(.dbd.O)2NHR.sup.7;
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O)2NHR.sup.7, substituted or
unsubstituted C.sub.2-C.sub.10heterocycloalkyl, or substituted or
unsubstituted heteroaryl; [0380] each R.sup.7 of
--NHC(.dbd.O)R.sup.7, --C(.dbd.O)NHR.sup.7, --NHS(.dbd.O)2R.sup.7,
--S(.dbd.O).sub.2NHR.sup.7; --NHC(.dbd.O)NHR.sup.7,
--NHS(.dbd.O).sub.2NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)R.sup.7,
--(C.sub.1-C.sub.3alkyl)-C(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O)2R.sup.7,
--(C.sub.1-C.sub.3alkyl)-S(.dbd.O)2NHR.sup.7;
--(C.sub.1-C.sub.3alky)-NHC(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O)2NHR.sup.7 is independently
selected from C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6heteroalkyl, a substituted or unsubstituted
C.sub.3-C.sub.10cycloalkyl, a substituted or unsubstituted
C.sub.2-C.sub.10heterocycloalkyl, a substituted or unsubstituted
aryl, a substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.10cycloalkyl), --C.sub.1-C.sub.6alkyl- (substituted
or unsubstituted C.sub.2-C.sub.10heterocycloalkyl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl),
--(CH2)p-CH(substituted or unsubstituted aryl)2,
--(CH.sub.2).sub.p--CH(substituted or unsubstituted heteroaryl)2,
--(CH.sub.2).sub.P--CH(substituted or unsubstituted
aryl)(substituted or unsubstituted heteroaryl), -(substituted or
unsubstituted aryl)-(substituted or unsubstituted aryl),
-(substituted or unsubstituted aryl)-(substituted or unsubstituted
heteroaryl), -(substituted or unsubstituted
heteroaryl)-(substituted or unsubstituted aryl), or -(substituted
or unsubstituted heteroaryl)-(substituted or unsubstituted
heteroaryl); [0381] p of R.sup.7 is selected from 0, 1 or 2; [0382]
R.sup.8a, R.sup.8b, R.sup.8c, and R.sup.8d of C(R.sup.8a)(R.sup.8b)
and C(R.sup.8c)(R.sup.8d) are independently selected from H,
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6fluoroalkyl, C.sub.1-C.sub.6
alkoxy, C.sub.1-C.sub.6heteroalkyl, and substituted or
unsubstituted aryl; [0383] or: [0384] R.sup.8a and R.sup.8d are as
defined above, and R.sup.8b and R.sup.8c together form a bond;
[0385] or: [0386] R.sup.8a and R.sup.8d are as defined above, and
R.sup.8b and R.sup.8c together with the atoms to which they are
attached form a substituted or unsubstituted fused 5-7 membered
saturated, or partially saturated carbocyclic ring or heterocyclic
ring comprising 1-3 heteroatoms selected from S, O and N, a
substituted or unsubstituted fused 5-10 membered aryl ring, or a
substituted or unsubstituted fused 5-10 membered heteroaryl ring
comprising 1-3 heteroatoms selected from S, O and N; [0387] or:
[0388] R.sup.8c and R.sup.8d are as defined above, and R.sup.8a and
R.sup.8b together with the atoms to which they are attached form a
substituted or unsubstituted saturated, or partially saturated 3-7
membered spirocycle or heterospirocycle comprising 1-3 heteroatoms
selected from S, O and N; [0389] or: [0390] R.sup.8a and R.sup.8b
are as defined above, and R.sup.8c and R.sup.8d together with the
atoms to which they are attached form a substituted or
unsubstituted saturated, or partially saturated 3-7 membered
spirocycle or heterospirocycle comprising 1-3 heteroatoms selected
from S, O and N; [0391] where each substituted alkyl, heteroalkyl,
fused ring, spirocycle, heterospirocycle, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl is substituted with 1-3
R.sup.9; and [0392] each R.sup.9 of R.sup.8a, R.sup.8b, R.sup.8c
and R.sup.8d is independently selected from halogen, --OH, --SH,
(C.dbd.O), CN, C.sub.1-C.sub.4alkyl, C.sub.1-C.sub.4fluoroalkyl,
C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 fluoroalkoxy, --NH.sub.2,
--NH(C.sub.1-C.sub.4alkyl), --NH(C.sub.1-C.sub.4alkyl).sub.2,
--C(.dbd.O)OH, --C(.dbd.0)NH.sub.2,
--C(.dbd.O)C.sub.1-C.sub.3alkyl, --S(.dbd.O).sub.2CH.sub.3,
--NH(C.sub.1-C.sub.4alkyl)-OH,
--NH(C.sub.1-C.sub.4alkyl)-O--(C-C.sub.4alkyl),
--O(C.sub.1-C.sub.4alkyl)-NH2;
--O(C.sub.1-C.sub.4alkyl)-NH--(C.sub.1-C.sub.4alkyl), and
--O(C.sub.1-C.sub.4alkyl)-N--(C.sub.1-C.sub.4alkyl).sub.2, or two
R.sup.9 together with the atoms to which they are attached form a
methylene dioxy or ethylene dioxy ring substituted or unsubstituted
with halogen, --OH, or C.sub.1-C.sub.3alkyl.
[0393] In any of the compounds described herein, the ILM can have
the structure of Formula (XLII), which are derived from the IAP
ligands described in WO Pub. No. 2013/071039, or an unnatural
mimetic thereof:
##STR00126##
wherein: [0394] W.sup.1 of Formula (XLII) is O, S, N--R.sup.A, or
C(R.sup.8a)(R.sup.8b); [0395] W.sup.2 of Formula (XLII) is O, S,
N--R.sup.A, or C(R.sup.8c)(R.sup.8d); provided that W.sup.1 and
W.sup.2 are not both O, or both S; [0396] R.sup.1 of Formula (XLII)
is selected from H, C.sub.1-C.sub.6alkyl,
C.sub.3-C.sub.6cycloalkyl, --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.3-C.sub.6cycloalkyl), substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl);
[0397] when X.sup.1 of Formula (XLII) is N--R.sup.A, then X.sup.2
is C.dbd.O, or CR.sup.2cR.sup.2d, and X.sup.3 is CR.sup.2aR.sup.2b;
[0398] or: [0399] when X.sup.1 of Formula (XLII) is selected from
S, S(O), or S(O).sub.2, then X.sup.2 is CR.sup.2cR.sup.2d, and
X.sup.3 is CR.sup.2aR.sup.2b. [0400] or: [0401] when X.sup.1 of
Formula (XLII) is O, then X.sup.2 is CR.sup.2cR.sup.2d and
N--R.sup.A and X.sup.3 is CR.sup.2aR.sup.2b; [0402] or: [0403] when
X.sup.1 of Formula (XLII) is CH.sub.3, then X.sup.2 is selected
from O, N--R.sup.A, S, S(O), or S(O).sub.2, and X.sup.3 is
CR.sup.2aR.sup.2b; [0404] when X.sup.1 of Formula (XLII) is
CR.sup.2eR.sup.2f and X.sup.2 is CR.sup.2cR.sup.2d, and R.sup.2e
and R.sup.2c together form a bond, and X.sup.3 of Formula (VLII) is
CR.sup.2aR.sup.2b; [0405] or: [0406] X.sup.1 and X.sup.3 of Formula
(XLII) are both CH.sub.2 and X.sup.2 of Formula (XLII) is C.dbd.0,
C.dbd.C(R.sup.C)2, or C.dbd.NR.sup.C; where each R.sup.C is
independently selected from H, --CN, --OH, alkoxy, substituted or
unsubstituted C1-C6alkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted heteroaryl);
[0407] or: [0408] X.sup.1 and X.sup.2 of Formula (XLII) are
independently selected from C and N, and are members of a fused
substituted or unsubstituted saturated or partially saturated 3-10
membered cycloalkyl ring, a fused substituted or unsubstituted
saturated or partially saturated 3-10 membered heterocycloalkyl
ring, a fused substituted or unsubstituted 5-10 membered aryl ring,
or a fused substituted or unsubstituted 5-10 membered heteroaryl
ring, and X.sup.3 is CR.sup.2aR.sup.2b. [0409] or: [0410] X.sup.2
and X.sup.3 of Formula (XLII) are independently selected from C and
N, and are members of a fused substituted or unsubstituted
saturated or partially saturated 3-10 membered cycloalkyl ring, a
fused substituted or unsubstituted saturated or partially saturated
3-10 membered heterocycloalkyl ring, a fused substituted or
unsubstituted 5-10 membered aryl ring, or a fused substituted or
unsubstituted 5-10 membered heteroaryl ring, and X.sup.1 of Formula
(VIII) is CR.sup.2eR.sup.2f; [0411] R.sup.A of N--R.sup.A is
selected from H, C.sub.1-C.sub.6alkyl,
--C(.dbd.O)C.sub.1-C.sub.2alkyl, substituted or unsubstituted aryl,
or substituted or unsubstituted heteroaryl; [0412] R.sup.2a,
R.sup.2b, R.sup.2c, R.sup.2d, R.sup.2e, and R.sup.2f of
CR.sup.2cR.sup.2d, CR.sup.2aR.sup.2b and CR.sup.2eR.sup.2f are
independently selected from H, substituted or unsubstituted
C1-C6alkyl, substituted or unsubstituted
C.sub.1-C.sub.6heteroalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl)
and --C(.dbd.O)R.sup.B; [0413] R.sup.B of --C(.dbd.O)R.sup.B is
selected from substituted or unsubstituted C.sub.1-C.sub.6alkyl,
substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted
or unsubstituted C.sub.2-C.sub.5heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl),
or --NR.sup.DR.sup.E; [0414] R.sup.D and R.sup.E of NR.sup.DR.sup.E
are independently selected from H, substituted or unsubstituted
C.sub.1-C.sub.6alkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, --C.sub.1-C.sub.6alkyl-
(substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted heteroaryl);
[0415] m of Formula (XLII) is selected from 0, 1 or 2; [0416] --U--
of Formula (XLII) is selected from --NHC(.dbd.O)--,
--C(.dbd.O)NH--, --NHS(.dbd.O).sub.2--, --S(.dbd.O).sub.2NH--,
--NHC(.dbd.O)NH--, --NH(C.dbd.O)O--, --O(C.dbd.O)NH--, or
--NHS(.dbd.O).sub.2NH--; [0417] R.sup.3 of Formula (XLII) is
selected from C.sub.1-C.sub.3alkyl, or C.sub.1-C.sub.3fluoroalkyl;
[0418] R.sup.4 of Formula (XLII) is selected from --NHR.sup.5,
--N(R.sup.5).sub.2, --N+(R.sup.5).sub.3 or --OR.sup.5; [0419] each
R.sup.5 of --NHR.sup.5, --N(R.sup.5).sub.2, --N+(R.sup.5).sub.3 and
--OR.sup.5 is independently selected from H, C.sub.1-C.sub.3alkyl,
C.sub.1-C.sub.3haloalkyl, C.sub.1-C.sub.3heteroalkyl and
--C.sub.1-C.sub.3alkyl-(C.sub.3-C.sub.5cycloalkyl); [0420] or:
[0421] R.sup.3 and R.sup.5 of Formula (XLII) together with the
atoms to which they are attached form a substituted or
unsubstituted 5-7 membered ring; [0422] or: [0423] R.sup.3 of
Formula (XLII) is bonded to a nitrogen atom of U to form a
substituted or unsubstituted 5-7 membered ring; [0424] R.sup.6 of
Formula (XLII) is selected from --NHC(.dbd.O)R,
--C(.dbd.O)NHR.sup.7, --NHS(.dbd.O)2R.sup.7,
--S(.dbd.O).sub.2NHR.sup.7; --NHC(.dbd.O)NHR.sup.7,
--NHS(.dbd.O).sub.2NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)R.sup.7,
--(C.sub.1-C.sub.3alkyl)-C(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O).sub.2R.sup.7,
--(C.sub.1-C.sub.3alkyl)-S(.dbd.O).sub.2NHR.sup.7;
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O).sub.2NHR.sup.7, substituted or
unsubstituted C.sub.2-C.sub.10heterocycloalkyl, or substituted or
unsubstituted heteroaryl; [0425] each R.sup.7 of
--NHC(.dbd.O)R.sup.7, --C(.dbd.O)NHR.sup.7, --NHS(.dbd.O)2R.sup.7,
--S(.dbd.O).sub.2NHR.sup.7; --NHC(.dbd.O)NHR.sup.7,
--NHS(.dbd.O).sub.2NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)R.sup.7,
--(C.sub.1-C.sub.3alkyl)-C(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O)2R.sup.7,
--(C.sub.1-C.sub.3alkyl)-S(.dbd.O)2NHR.sup.7;
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O)2NHR.sup.7 is independently
selected from C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6heteroalkyl, a substituted or unsubstituted
C3-C10cycloalkyl, a substituted or unsubstituted
C.sub.2-C.sub.10heterocycloalkyl, a substituted or unsubstituted
aryl, a substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.10cycloalkyl), --C.sub.1-C.sub.6alkyl- (substituted
or unsubstituted C.sub.2-C.sub.10heterocycloalkyl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl),
--(CH2)p-CH(substituted or unsubstituted aryl)2,
--(CH.sub.2).sub.p--CH(substituted or unsubstituted heteroaryl)2,
--(CH.sub.2).sub.P--CH(substituted or unsubstituted
aryl)(substituted or unsubstituted heteroaryl), -(substituted or
unsubstituted aryl)-(substituted or unsubstituted aryl),
-(substituted or unsubstituted aryl)-(substituted or unsubstituted
heteroaryl), -(substituted or unsubstituted
heteroaryl)-(substituted or unsubstituted aryl), or -(substituted
or unsubstituted heteroaryl)-(substituted or unsubstituted
heteroaryl); [0426] p of R.sup.7 is selected from 0, 1 or 2; [0427]
R.sup.8a, R.sup.8b, R.sup.8c, and R.sup.8d of C(R.sup.8a)(R.sup.8b)
and C(R.sup.8c)(R.sup.8d) are independently selected from H,
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6fluoroalkyl, C.sub.1-C.sub.6
alkoxy, C.sub.1-C.sub.6heteroalkyl, and substituted or
unsubstituted aryl; [0428] or: [0429] R.sup.8a and R.sup.8d are as
defined above, and R.sup.8b and R.sup.8c together form a bond;
[0430] or: [0431] R.sup.8a and R.sup.8d are as defined above, and
R.sup.8b and R.sup.8c together with the atoms to which they are
attached form a substituted or unsubstituted fused 5-7 membered
saturated, or partially saturated carbocyclic ring or heterocyclic
ring comprising 1-3 heteroatoms selected from S, O and N, a
substituted or unsubstituted fused 5-10 membered aryl ring, or a
substituted or unsubstituted fused 5-10 membered heteroaryl ring
comprising 1-3 heteroatoms selected from S, O and N; [0432] or:
[0433] R.sup.8c and R.sup.8d are as defined above, and R.sup.8a and
R.sup.8b together with the atoms to which they are attached form a
substituted or unsubstituted saturated, or partially saturated 3-7
membered spirocycle or heterospirocycle comprising 1-3 heteroatoms
selected from S, O and N; [0434] or: [0435] R.sup.8a and R.sup.8b
are as defined above, and R.sup.8c and R.sup.8d together with the
atoms to which they are attached form a substituted or
unsubstituted saturated, or partially saturated 3-7 membered
spirocycle or heterospirocycle comprising 1-3 heteroatoms selected
from S, O and N; [0436] where each substituted alkyl, heteroalkyl,
fused ring, spirocycle, heterospirocycle, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl is substituted with 1-3
R.sup.9; and [0437] each R.sup.9 of R.sup.8a, R.sup.8b, R.sup.8c
and R.sup.8d is independently selected from halogen, --OH, --SH,
(C.dbd.O), CN, C.sub.1-C.sub.4alkyl, C1-C4fluoroalkyl,
C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 fluoroalkoxy, --NH.sub.2,
--NH(C.sub.1-C.sub.4alkyl), --NH(C.sub.1-C.sub.4alkyl).sub.2,
--C(.dbd.O)OH, --C(.dbd.O)NH.sub.2,
--C(.dbd.O)C.sub.1-C.sub.3alkyl, --S(.dbd.O).sub.2CH.sub.3,
--NH(C.sub.1-C.sub.4alkyl)-OH,
--NH(C.sub.1-C.sub.4alkyl)-O--(C-C.sub.4alkyl),
--O(C.sub.1-C.sub.4alkyl)-NH2;
--O(C.sub.1-C.sub.4alkyl)-NH--(C.sub.1-C.sub.4alkyl), and
--O(C.sub.1-C.sub.4alkyl)-N--(C.sub.1-C.sub.4alkyl).sub.2, or two
R.sup.9 together with the atoms to which they are attached form a
methylene dioxy or ethylene dioxy ring substituted or unsubstituted
with halogen, --OH, or C.sub.1-C.sub.3alkyl.
[0438] In any of the compounds described herein, the ILM can have
the structure of Formula (XLIII), which is derived from the IAP
ligands described in WO Pub. No. 2013/071039, or an unnatural
mimetic thereof:
##STR00127##
wherein: [0439] W.sup.1 of Formula (XLIII) is selected from O, S,
N--R.sup.A, or C(R.sup.8a)(R.sup.8b); [0440] W.sup.2 of Formula
(XLIII) is selected from O, S, N--R.sup.A, or
C(R.sup.8c)(R.sup.8d); provided that W.sup.1 and W.sup.2 are not
both O, or both S; [0441] R.sup.1 of Formula (XLIII) is selected
from H, C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.6cycloalkyl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl);
[0442] when X.sup.1 of Formula (XLIII) is selected from N--R.sup.A,
S, S(O), or S(O).sub.2, then X.sup.2 of Formula (XLIII) is
CR.sup.2cR.sup.2d, and X.sup.3 of Formula (XLIII) is
CR.sup.2aR.sup.2b; [0443] or: [0444] when X.sup.1 of Formula
(XLIII) is O, then X.sup.2 of Formula (XLIII) is selected from O,
N--R.sup.A, S, S(O), or S(O).sub.2, and X.sup.3 of Formula (XLIII)
is CR.sup.2aR.sup.2b; [0445] or: [0446] when X.sup.1 of Formula
(XLIII) is CR.sup.2eR.sup.2f and X.sup.2 of Formula (XLIII) is
CR.sup.2cR.sup.2d, and R.sup.2e and R.sup.2c together form a bond,
and X.sup.3 of Formula (XLIII) is CR.sup.2aR.sup.2b; [0447] or:
[0448] X.sup.1 and X.sup.2 of Formula (XLIII) are independently
selected from C and N, and are members of a fused substituted or
unsubstituted saturated or partially saturated 3-10 membered
cycloalkyl ring, a fused substituted or unsubstituted saturated or
partially saturated 3-10 membered heterocycloalkyl ring, a fused
substituted or unsubstituted 5-10 membered aryl ring, or a fused
substituted or unsubstituted 5-10 membered heteroaryl ring, and
X.sup.3 of Formula (XLIII) is CR.sup.2aR.sup.2b; [0449] or: [0450]
X.sup.2 and X.sup.3 of Formula (XLIII) are independently selected
from C and N, and are members of a fused substituted or
unsubstituted saturated or partially saturated 3-10 membered
cycloalkyl ring, a fused substituted or unsubstituted saturated or
partially saturated 3-10 membered heterocycloalkyl ring, a fused
substituted or unsubstituted 5-10 membered aryl ring, or a fused
substituted or unsubstituted 5-10 membered heteroaryl ring, and
X.sup.1 of Formula (VLII) is CR.sup.2eR.sup.2f; [0451] R.sup.A of
N--R.sup.A is H, C.sub.1-C.sub.6alkyl,
--C(.dbd.O)C.sub.1-C.sub.2alkyl, substituted or unsubstituted aryl,
or substituted or unsubstituted heteroaryl; [0452] R.sup.2a,
R.sup.2b, R.sup.2c, R.sup.2d, R.sup.2e, and R.sup.2f of
CR.sup.2cR.sup.2d, CR.sup.2aR.sup.2b and CR.sup.2eR.sup.2f are
independently selected from H, substituted or unsubstituted
C.sub.1-C.sub.6alkyl, substituted or unsubstituted
C.sub.1-C.sub.6heteroalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl)
and --C(.dbd.O)R.sup.B; [0453] R.sup.B of --C(.dbd.O)R.sup.B is
substituted or unsubstituted C.sub.1-C.sub.6alkyl, substituted or
unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl),
or --NR.sup.DR.sup.E; [0454] R.sup.D and R.sup.E of NR.sup.DR.sup.E
are independently selected from H, substituted or unsubstituted
C.sub.1-C.sub.6alkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, --C.sub.1-C.sub.6alkyl-
(substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted heteroaryl);
[0455] m of Formula (XLIII) is 0, 1 or 2; [0456] --U-- of Formula
(XLIII) is --NHC(.dbd.O)--, --C(.dbd.O)NH--, --NHS(.dbd.O).sub.2--,
--S(.dbd.O).sub.2NH--, --NHC(.dbd.O)NH--, --NH(C.dbd.O)O--,
--O(C.dbd.O)NH--, or --NHS(.dbd.O).sub.2NH--; [0457] R.sup.3 of
Formula (XLIII) is C.sub.1-C.sub.3alkyl, or
C.sub.1-C.sub.3fluoroalkyl; [0458] R.sup.4 of Formula (XLIII) is
--NHR.sup.5, --N(R.sup.5).sub.2, --N+(R.sup.5).sub.3 or --OR.sup.5;
[0459] each R.sup.5 of --NHR.sup.5, --N(R.sup.5).sub.2,
--N+(R.sup.5).sub.3 and --OR.sup.5 is independently selected from
H, C.sub.1-C.sub.3alkyl, C.sub.1-C.sub.3haloalkyl,
C.sub.1-C.sub.3heteroalkyl and
--C.sub.1-C.sub.3alkyl-(C.sub.3-C.sub.5cycloalkyl); [0460] or:
[0461] R.sup.3 and R.sup.5 of Formula (XLIII) together with the
atoms to which they are attached form a substituted or
unsubstituted 5-7 membered ring; [0462] or: [0463] R.sup.3 of
Formula (XLIII) is bonded to a nitrogen atom of U to form a
substituted or unsubstituted 5-7 membered ring; [0464] R.sup.6 of
Formula (XLIII) is selected from --NHC(.dbd.O)R.sup.7,
--C(.dbd.O)NHR.sup.7, --NHS(.dbd.O)2R.sup.7,
--S(.dbd.O).sub.2NHR.sup.7; --NHC(.dbd.O)NHR.sup.7,
--NHS(.dbd.O).sub.2NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)R.sup.7,
--(C.sub.1-C.sub.3alkyl)-C(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O).sub.2R.sup.7,
--(C.sub.1-C.sub.3alkyl)-S(.dbd.O).sub.2NHR.sup.7;
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O).sub.2NHR.sup.7, substituted or
unsubstituted C.sub.2-C.sub.10heterocycloalkyl, or substituted or
unsubstituted heteroaryl; [0465] each R.sup.7 of
--NHC(.dbd.O)R.sup.7, --C(.dbd.O)NHR.sup.7, --NHS(.dbd.O)2R.sup.7,
--S(.dbd.O).sub.2NHR.sup.7; --NHC(.dbd.O)NHR.sup.7,
--NHS(.dbd.O).sub.2NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)R.sup.7,
--(C.sub.1-C.sub.3alkyl)-C(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O)2R.sup.7,
--(C.sub.1-C.sub.3alkyl)-S(.dbd.O)2NHR.sup.7;
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O)2NHR.sup.7 is independently
selected from C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6heteroalkyl, a substituted or unsubstituted
C3-C10cycloalkyl, a substituted or unsubstituted
C.sub.2-C.sub.10heterocycloalkyl, a substituted or unsubstituted
aryl, a substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.10cycloalkyl), --C.sub.1-C.sub.6alkyl- (substituted
or unsubstituted C.sub.2-C.sub.10heterocycloalkyl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl),
--(CH2)p-CH(substituted or unsubstituted aryl)2,
--(CH.sub.2).sub.p--CH(substituted or unsubstituted heteroaryl)2,
--(CH.sub.2).sub.P--CH(substituted or unsubstituted
aryl)(substituted or unsubstituted heteroaryl), -(substituted or
unsubstituted aryl)-(substituted or unsubstituted aryl),
-(substituted or unsubstituted aryl)-(substituted or unsubstituted
heteroaryl), -(substituted or unsubstituted
heteroaryl)-(substituted or unsubstituted aryl), or -(substituted
or unsubstituted heteroaryl)-(substituted or unsubstituted
heteroaryl); [0466] p of R.sup.7 is 0, 1 or 2; [0467] R.sup.8a,
R.sup.8b, R.sup.8c, and R.sup.8d of C(R.sup.8a)(R.sup.8b) and
C(R.sup.8c)(R.sup.8d) are independently selected from H,
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6fluoroalkyl, C.sub.1-C.sub.6
alkoxy, C.sub.1-C.sub.6heteroalkyl, and substituted or
unsubstituted aryl; [0468] or: [0469] R.sup.8a and R.sup.8d are as
defined above, and R.sup.8b and R.sup.8c together form a bond;
[0470] or: [0471] R.sup.8a and R.sup.8d are as defined above, and
R.sup.8b and R.sup.8c together with the atoms to which they are
attached form a substituted or unsubstituted fused 5-7 membered
saturated, or partially saturated carbocyclic ring or heterocyclic
ring comprising 1-3 heteroatoms selected from S, O and N, a
substituted or unsubstituted fused 5-10 membered aryl ring, or a
substituted or unsubstituted fused 5-10 membered heteroaryl ring
comprising 1-3 heteroatoms selected from S, O and N; [0472] or:
[0473] R.sup.8c and R.sup.8d are as defined above, and R.sup.8a and
R.sup.8b together with the atoms to which they are attached form a
substituted or unsubstituted saturated, or partially saturated 3-7
membered spirocycle or heterospirocycle comprising 1-3 heteroatoms
selected from S, O and N; [0474] or: [0475] R.sup.8a and R.sup.8b
are as defined above, and R.sup.8c and R.sup.8d together with the
atoms to which they are attached form a substituted or
unsubstituted saturated, or partially saturated 3-7 membered
spirocycle or heterospirocycle comprising 1-3 heteroatoms selected
from S, O and N; [0476] where each substituted alkyl, heteroalkyl,
fused ring, spirocycle, heterospirocycle, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl is substituted with 1-3
R.sup.9; and [0477] each R.sup.9 of R.sup.8a, R.sup.8b, R.sup.8c
and R.sup.8d is independently selected from halogen, --OH, --SH,
(C.dbd.O), CN, C.sub.1-C.sub.4alkyl, C.sub.1-C.sub.4fluoroalkyl,
C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 fluoroalkoxy, --NH.sub.2,
--NH(C.sub.1-C.sub.4alkyl), --NH(C.sub.1-C.sub.4alkyl).sub.2,
--C(.dbd.O)OH, --C(.dbd.O)NH.sub.2,
--C(.dbd.O)C.sub.1-C.sub.3alkyl, --S(.dbd.O).sub.2CH.sub.3,
--NH(C.sub.1-C.sub.4alkyl)-OH,
--NH(C.sub.1-C.sub.4alkyl)-O--(C-C.sub.4alkyl),
--O(C.sub.1-C.sub.4alkyl)-NH2;
--O(C.sub.1-C.sub.4alkyl)-NH--(C.sub.1-C.sub.4alkyl), and
--O(C.sub.1-C.sub.4alkyl)-N--(C.sub.1-C.sub.4alkyl).sub.2, or two
R.sup.9 together with the atoms to which they are attached form a
methylene dioxy or ethylene dioxy ring substituted or unsubstituted
with halogen, --OH, or C.sub.1-C.sub.3alkyl.
[0478] In any of the compounds described herein, the ILM can have
the structure of Formula (XLIV), which is derived from the IAP
ligands described in WO Pub. No. 2013/071039, or an unnatural
mimetic thereof:
##STR00128##
wherein: [0479] W.sup.1 of Formula (XLIV) is selected from O, S,
N--R.sup.A, or C(R.sup.8a)(R.sup.8b); [0480] W.sup.2 of Formula
(XLIV) is selected from O, S, N--R.sup.A, or C(R.sup.8c)(R.sup.8d);
provided that W.sup.1 and W.sup.2 are not both O, or both S; [0481]
W.sup.3 of Formula (XLIV) is selected from O, S, N--R.sup.A, or
C(R.sup.8e)(R.sup.8f), providing that the ring comprising W.sup.1,
W.sup.2, and W.sup.3 does not comprise two adjacent oxygen atoms or
sulfer atoms; [0482] R.sup.1 of Formula (XLIV) is selected from H,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.6cycloalkyl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl);
[0483] when X.sup.1 of Formula (XLIV) is O, then X.sup.2 of Formula
(XLIV) is selected from CR.sup.2cR.sup.2d and N--R.sup.A, and
X.sup.3 of Formula (XLIV) is CR.sup.2aR.sup.2b; [0484] or: [0485]
when X.sup.1 of Formula (XLIV) is CH.sub.2, then X.sup.2 of Formula
(XLIV) is selected from O, N--R.sup.A, S, S(O), or S(O).sub.2, and
X.sup.3 of Formula (XLIV) is CR.sup.2aR.sup.2b; [0486] or: [0487]
when X.sup.1 of Formula (XLIV) is CR.sup.2eR.sup.2f and X.sup.2 of
Formula (XLIV) is CR.sup.2cR.sup.2d, and R.sup.2e and R.sup.2c
together form a bond, and X.sup.3 of Formula (VLIV) is
CR.sup.2aR.sup.2b; [0488] or: [0489] X.sup.1 and X.sup.3 of Formula
(XLIV) are both CH.sub.2 and X.sup.2 of Formula (XLII) is C.dbd.0,
C.dbd.C(R.sup.C)2, or C.dbd.NR.sup.C; where each R.sup.C is
independently selected from H, --CN, --OH, alkoxy, substituted or
unsubstituted C.sub.1-C.sub.6alkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted heteroaryl);
[0490] or: [0491] X.sup.1 and X.sup.2 of Formula (XLIV) are
independently selected from C and N, and are members of a fused
substituted or unsubstituted saturated or partially saturated 3-10
membered cycloalkyl ring, a fused substituted or unsubstituted
saturated or partially saturated 3-10 membered heterocycloalkyl
ring, a fused substituted or unsubstituted 5-10 membered aryl ring,
or a fused substituted or unsubstituted 5-10 membered heteroaryl
ring, and X.sup.3 of Formula (XLIV) is CR.sup.2aR.sup.2b; [0492]
or: [0493] X.sup.2 and X.sup.3 of Formula (XLIV) are independently
selected from C and N, and are members of a fused substituted or
unsubstituted saturated or partially saturated 3-10 membered
cycloalkyl ring, a fused substituted or unsubstituted saturated or
partially saturated 3-10 membered heterocycloalkyl ring, a fused
substituted or unsubstituted 5-10 membered aryl ring, or a fused
substituted or unsubstituted 5-10 membered heteroaryl ring, and
X.sup.1 of Formula (VLIV) is CR.sup.2eR.sup.2f; [0494] R.sup.A of
N--R.sup.A is selected from H, C.sub.1-C.sub.6alkyl,
--C(.dbd.O)C.sub.1-C.sub.2alkyl, substituted or unsubstituted aryl,
or substituted or unsubstituted heteroaryl; [0495] R.sup.2a,
R.sup.2b, R.sup.2c, R.sup.2d, R.sup.2e, and R.sup.2f of
CR.sup.2cR.sup.2d, CR.sup.2aR.sup.2b and CR.sup.2eR.sup.2f are
independently selected from H, substituted or unsubstituted
C.sub.1-C.sub.6alkyl, substituted or unsubstituted
C.sub.1-C.sub.6heteroalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl)
and --C(.dbd.O)R.sup.B; [0496] R.sup.B of --C(.dbd.O)R.sup.B is
selected from substituted or unsubstituted C.sub.1-C.sub.6alkyl,
substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted
or unsubstituted C.sub.2-C.sub.5heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), --C.sub.1-C.sub.6alkyl-(substituted or
unsubstituted C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl),
or --NR.sup.DR.sup.E; [0497] R.sup.D and R.sup.E of NR.sup.DR.sup.E
are independently selected from H, substituted or unsubstituted
C.sub.1-C.sub.6alkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, --C.sub.1-C.sub.6alkyl-
(substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.2-C.sub.5heterocycloalkyl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl), or
--C.sub.1-C.sub.6alkyl- (substituted or unsubstituted heteroaryl);
m of Formula (XLIV) is selected from 0, 1 or 2; [0498] --U-- of
Formula (XLIV) is selected from --NHC(.dbd.O)--, --C(.dbd.O)NH--,
--NHS(.dbd.O).sub.2--, --S(.dbd.O).sub.2NH--, --NHC(.dbd.O)NH--,
--NH(C.dbd.O)O--, --O(C.dbd.O)NH--, or --NHS(.dbd.O).sub.2NH--;
[0499] R.sup.3 of Formula (XLIV) is selected from
C.sub.1-C.sub.3alkyl, or C.sub.1-C.sub.3fluoroalkyl; [0500] R.sup.4
of Formula (XLIV) is selected from --NHR.sup.5, --N(R.sup.5).sub.2,
--N+(R.sup.5).sub.3 or --OR.sup.5; [0501] each R.sup.5 of
--NHR.sup.5, --N(R.sup.5).sub.2, --N+(R.sup.5).sub.3 and --OR.sup.5
is independently selected from H, C.sub.1-C.sub.3alkyl,
C.sub.1-C.sub.3haloalkyl, C.sub.1-C.sub.3heteroalkyl and
--C.sub.1-C.sub.3alkyl-(C.sub.3-C.sub.5cycloalkyl); [0502] or:
[0503] R.sup.3 and R.sup.5 of Formula (XLIV) together with the
atoms to which they are attached form a substituted or
unsubstituted 5-7 membered ring; [0504] or: [0505] R.sup.3 of
Formula (XLIII) is bonded to a nitrogen atom of U to form a
substituted or unsubstituted 5-7 membered ring; [0506] R.sup.6 of
Formula (XLIII) is selected from --NHC(.dbd.O)R.sup.7,
--C(.dbd.O)NHR.sup.7, --NHS(.dbd.O)2R.sup.7,
--S(.dbd.O).sub.2NHR.sup.7; --NHC(.dbd.O)NHR.sup.7,
--NHS(.dbd.O).sub.2NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)R.sup.7,
--(C.sub.1-C.sub.3alkyl)-C(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O).sub.2R.sup.7,
--(C.sub.1-C.sub.3alkyl)-S(.dbd.O).sub.2NHR.sup.7;
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O).sub.2NHR.sup.7, substituted or
unsubstituted C.sub.2-C.sub.10heterocycloalkyl, or substituted or
unsubstituted heteroaryl; [0507] each R.sup.7 of
--NHC(.dbd.O)R.sup.7, --C(.dbd.O)NHR.sup.7, --NHS(.dbd.O)2R.sup.7,
--S(.dbd.O).sub.2NHR.sup.7; --NHC(.dbd.O)NHR.sup.7,
--NHS(.dbd.O).sub.2NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)R.sup.7,
--(C.sub.1-C.sub.3alkyl)-C(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O)2R.sup.7,
--(C.sub.1-C.sub.3alkyl)-S(.dbd.O)2NHR.sup.7;
--(C.sub.1-C.sub.3alkyl)-NHC(.dbd.O)NHR.sup.7,
--(C.sub.1-C.sub.3alkyl)-NHS(.dbd.O)2NHR.sup.7 is independently
selected from C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6heteroalkyl, a substituted or unsubstituted
C3-C10cycloalkyl, a substituted or unsubstituted
C.sub.2-C.sub.10heterocycloalkyl, a substituted or unsubstituted
aryl, a substituted or unsubstituted heteroaryl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted
C.sub.3-C.sub.10cycloalkyl), --C.sub.1-C.sub.6alkyl- (substituted
or unsubstituted C.sub.2-C.sub.10heterocycloalkyl,
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted aryl),
--C.sub.1-C.sub.6alkyl-(substituted or unsubstituted heteroaryl),
--(CH2)p-CH(substituted or unsubstituted aryl)2,
--(CH.sub.2).sub.p--CH(substituted or unsubstituted heteroaryl)2,
--(CH.sub.2).sub.P--CH(substituted or unsubstituted
aryl)(substituted or unsubstituted heteroaryl), -(substituted or
unsubstituted aryl)-(substituted or unsubstituted aryl),
-(substituted or unsubstituted aryl)-(substituted or unsubstituted
heteroaryl), -(substituted or unsubstituted
heteroaryl)-(substituted or unsubstituted aryl), or -(substituted
or unsubstituted heteroaryl)-(substituted or unsubstituted
heteroaryl); [0508] p of R.sup.7 is selected from 0, 1 or 2; [0509]
R.sup.8a, R.sup.8b, R.sup.8c, R.sup.8d, R.sup.8e, and R.sup.8f of
C(R.sup.8a)(R.sup.8b), C(R.sup.8c)(R.sup.8d) and
C(R.sup.8e)(R.sup.8f) are independently selected from H,
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6fluoroalkyl, C.sub.1-C.sub.6
alkoxy, C.sub.1-C.sub.6heteroalkyl, and substituted or
unsubstituted aryl; [0510] or: [0511] R.sup.8a, R.sup.8d, R.sup.8e,
and R.sup.8f of C(R.sup.8a)(R.sup.8b), C(R.sup.8c)(R.sup.8d) and
C(R.sup.8e)(R.sup.8f) are as defined above, and R.sup.8b and
R.sup.8c together form a bond; [0512] or: [0513] R.sup.8a,
R.sup.8b, R.sup.8d, and R.sup.8f of C(R.sup.8a)(R.sup.8b),
C(R.sup.8c)(R.sup.8d) and C(R.sup.8e)(R.sup.8f) are as defined
above, and R.sup.8c and R.sup.8e together form a bond; [0514] or:
[0515] R.sup.8a, R.sup.8d, R.sup.8e, and R.sup.8f of
C(R.sup.8a)(R.sup.8b), C(R.sup.8c)(R.sup.8d) and
C(R.sup.8e)(R.sup.8f) are as defined above, and R.sup.8b and
R.sup.8c together with the atoms to which they are attached form a
substituted or unsubstituted fused 5-7 membered saturated, or
partially saturated carbocyclic ring or heterocyclic ring
comprising 1-3 heteroatoms selected from S, O and N, a substituted
or unsubstituted fused 5-10 membered aryl ring, or a substituted or
unsubstituted fused 5-10 membered heteroaryl ring comprising 1-3
heteroatoms selected from S, O and N; [0516] or: [0517] R.sup.8a,
R.sup.8b, R.sup.8d, and R.sup.8f of C(R.sup.8a)(R.sup.8b),
C(R.sup.8c)(R.sup.8d) and C(R.sup.8e)(R.sup.8f) are as defined
above, and R.sup.8c and R.sup.8e together with the atoms to which
they are attached form a substituted or unsubstituted fused 5-7
membered saturated, or partially saturated carbocyclic ring or
heterocyclic ring comprising 1-3 heteroatoms selected from S, O and
N, a substituted or unsubstituted fused 5-10 membered aryl ring, or
a substituted or unsubstituted fused 5-10 membered heteroaryl ring
comprising 1-3 heteroatoms selected from S, O and N; [0518] or:
[0519] R.sup.8c, R.sup.8d, R.sup.8e, and R.sup.8f of
C(R.sup.8c)(R.sup.8d) and C(R.sup.8e)(R.sup.8f) are as defined
above, and R.sup.8a and R.sup.8b together with the atoms to which
they are attached form a substituted or unsubstituted saturated, or
partially saturated 3-7 membered spirocycle or heterospirocycle
comprising 1-3 heteroatoms selected from S, O and N; [0520] or:
[0521] R.sup.8a, R.sup.8b, R.sup.8e, and R.sup.8f of
C(R.sup.8a)(R.sup.8b) and C(R.sup.8e)(R.sup.8f) are as defined
above, and R.sup.8c and R.sup.8d together with the atoms to which
they are attached form a substituted or unsubstituted saturated, or
partially saturated 3-7 membered spirocycle or heterospirocycle
comprising 1-3 heteroatoms selected from S, O and N; [0522] or:
[0523] R.sup.8a, R.sup.8b, R.sup.8c, and R.sup.8d of
C(R.sup.8a)(R.sup.8b) and C(R.sup.8c)(R.sup.8d) are as defined
above, and R.sup.8e and R.sup.8f together with the atoms to which
they are attached form a substituted or unsubstituted saturated, or
partially saturated 3-7 membered spirocycle or heterospirocycle
comprising 1-3 heteroatoms selected from S, O and N; [0524] or:
[0525] where each substituted alkyl, heteroalkyl, fused ring,
spirocycle, heterospirocycle, cycloalkyl, heterocycloalkyl, aryl or
heteroaryl is substituted with 1-3 R.sup.9; and [0526] each R.sup.9
of R.sup.8a, R.sup.8b, R.sup.8c, R.sup.8d, R.sup.8e, and R.sup.8f
is independently selected from halogen, --OH, --SH, (C.dbd.O), CN,
C.sub.1-C.sub.4alkyl, C1-C4fluoroalkyl, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 fluoroalkoxy, --NH.sub.2,
--NH(C.sub.1-C.sub.4alkyl), --NH(C.sub.1-C.sub.4alkyl).sub.2,
--C(.dbd.O)OH, --C(.dbd.O)NH.sub.2,
--C(.dbd.O)C.sub.1-C.sub.3alkyl, --S(.dbd.O).sub.2CH.sub.3,
--NH(C.sub.1-C.sub.4alkyl)-OH,
--NH(C.sub.1-C.sub.4alkyl)-O--(C-C.sub.4alkyl),
--O(C.sub.1-C.sub.4alkyl)-NH2;
--O(C.sub.1-C.sub.4alkyl)-NH--(C.sub.1-C.sub.4alkyl), and
--O(C.sub.1-C.sub.4alkyl)-N--(C.sub.1-C.sub.4alkyl).sub.2, or two
R.sup.9 together with the atoms to which they are attached form a
methylene dioxy or ethylene dioxy ring substituted or unsubstituted
with halogen, --OH, or C.sub.1-C.sub.3alkyl.
[0527] In any of the compounds described herein, the ILM can have
the structure of Formula (XLV), (XLVI) or (XLVII), which is derived
from the IAP ligands described in Vamos, M., et al., Expedient
synthesis of highly potent antagonists of inhibitor of apoptosis
proteins (IAPs) with unique selectivity for ML-IAP, ACS Chem.
Biol., 8(4), 725-32 (2013), or an unnatural mimetic thereof:
##STR00129##
wherein: [0528] R.sup.2, R.sup.3 and R.sup.4 of Formula (XLV) are
independently selected from H or ME; [0529] X of Formula (XLV) is
independently selected from O or S; and [0530] R.sup.1 of Formula
(XLV) is selected from:
##STR00130##
[0531] In a particular embodiment, the ILM has a structure
according to Formula (XLVIII):
##STR00131##
wherein R.sup.3 and R.sup.4 of Formula (XLVIII) are independently
selected from H or ME;
##STR00132##
is a 5-member heterocycle selected from:
##STR00133##
[0532] In a particular embodiment, the
##STR00134##
of Formula XLVIII) is
##STR00135##
[0534] In a particular embodiment, the ILM has a structure and
attached to a linker group L as shown below:
##STR00136##
[0535] In a particular embodiment, the ILM has a structure
according to Formula (XLIX), (L), or (LI):
##STR00137##
wherein: R.sup.3 of Formula (XLIX), (L) or (LI) are independently
selected from H or ME;
##STR00138##
is a 5-member heterocycle selected from:
##STR00139##
and L of Formula (XLIX), (L) or (LI) is selected from:
##STR00140##
[0536] In a particular embodiment, L of Formula (XLIX), (L), or
(LI)
##STR00141##
[0537] In a particular embodiment, the ILM has a structure
according to Formula (LII):
##STR00142##
[0538] In a particular embodiment, the ILM according to Formula
(LII) is chemically linked to the linker group L in the area
denoted with
##STR00143##
and as shown below:
##STR00144##
[0539] In any of the compounds described herein, the ILM can have
the structure of Formula (LIII) or (LIV), which is based on the IAP
ligands described in Hennessy, E J, et al., Discovery of
aminopiperidine-based Smac mimetics as IAP antagonists, Bioorg.
Med. Chem. Lett., 22(4), 1960-4 (2012), or an unnatural mimetic
thereof:
##STR00145##
wherein:
[0540] R.sup.1 of Formulas (LIII) and (LIV) is selected from:
##STR00146##
[0541] R.sup.2 of Formulas (LIII) and (LIV) is selected from H or
Me;
[0542] R.sup.3 of Formulas (LIII) and (LIV) is selected from:
##STR00147##
[0543] X of is selected from H, halogen, methyl, methoxy, hydroxy,
nitro or trifluoromethyl.
[0544] In any of the compounds described herein, the ILM can have
the structure of and be chemically linked to the linker as shown in
Formula (LV) or (LVI), or an unnatural mimetic thereof:
##STR00148##
[0545] In any of the compounds described herein, the ILM can have
the structure of Formula (LVII), which is based on the IAP ligands
described in Cohen, F, et al., Orally bioavailable antagonists of
inhibitor of apoptosis proteins based on an azabicyclooctane
scaffold, J. Med. Chem., 52(6), 1723-30 (2009), or an unnatural
mimetic thereof:
##STR00149##
wherein:
[0546] R1 of Formulas (LVII) is selected from:
##STR00150##
[0547] X of
##STR00151##
is selected from H, fluoro, methyl or methoxy.
[0548] In a particular embodiment, the ILM is represented by the
following structure:
##STR00152##
[0549] In a particular embodiment, the ILM is selected from the
group consisting of, and which the chemical link between the ILM
and linker group L is shown:
##STR00153##
[0550] In any of the compounds described herein, the ILM is
selected from the group consisting of the structures below, which
are based on the IAP ligands described in Asano, M, et al., Design,
sterioselective synthesis, and biological evaluation of novel
tri-cyclic compounds as inhibitor of apoptosis proteins (IAP)
antagonists, Bioorg. Med. Chem., 21(18): 5725-37 (2013), or an
unnatural mimetic thereof:
##STR00154##
[0551] In a particular embodiment, the ILM is selected from the
group consisting of, and which the chemical link between the ILM
and linker group L is shown:
##STR00155##
[0552] In any of the compounds described herein, the ILM can have
the structure of Formula (LVIII), which is based on the IAP ligands
described in Asano, M, et al., Design, sterioselective synthesis,
and biological evaluation of novel tri-cyclic compounds as
inhibitor of apoptosis proteins (IAP) antagonists, Bioorg. Med.
Chem., 21(18): 5725-37 (2013), or an unnatural mimetic thereof:
##STR00156##
wherein X of Formula (LVIII) is one or two substituents
independently selected from H, halogen or cyano.
[0553] In any of the compounds described herein, the ILM can have
the structure of and be chemically linked to the linker group L as
shown in Formula (LIX) or (LX), or an unnatural mimetic
thereof:
##STR00157##
wherein X of Formula (LIX) and (LX) is one or two substituents
independently selected from H, halogen or cyano, and; and L of
Formulas (LIX) and (LX) is a linker group as described herein.
[0554] In any of the compounds described herein, the ILM can have
the structure of Formula (LXI), which is based on the IAP ligands
described in Ardecky, R J, et al., Design, synthesis and evaluation
of inhibitor of apoptosis (IAP) antagonists that are highly
selective for the BIR2 domain of XIAP, Bioorg. Med. Chem., 23(14):
4253-7 (2013), or an unnatural mimetic thereof:
##STR00158##
wherein:
##STR00159##
of Formula (LXI) is a natural or unnatural amino acid; and R.sup.2
of Formula (LXI) is selected from:
##STR00160##
[0555] In any of the compounds described herein, the ILM can have
the structure of and be chemically linked to the linker group L as
shown in Formula (LXII) or (LLXIII), or an unnatural mimetic
thereof:
##STR00161##
of Formula (LXI) is a natural or unnatural amino acid; and L of
Formula (LXI) is a linker group as described herein.
[0556] In any of the compounds described herein, the ILM can have
the structure selected from the group consisting of, which is based
on the IAP ligands described in Wang, J, et al., Discovery of novel
second mitochondrial-derived activator of caspase mimetics as
selective inhibitor or apoptosis protein inhibitors, J. Pharmacol.
Exp. Ther., 349(2): 319-29 (2014), or an unnatural mimetic
thereof:
##STR00162##
[0557] In any of the compounds described herein, the ILM has a
structure according to Formula (LXIX), which is based on the IAP
ligands described in Hird, A W, et al., Structure-based design and
synthesis of tricyclic IAP (Inhibitors of Apoptosis Proteins)
inhibitors, Bioorg. Med. Chem. Lett., 24(7): 1820-4 (2014), or an
unnatural mimetic thereof:
##STR00163##
wherein R of Formula LIX is selected from the group consisting
of:
##STR00164##
R1 of
##STR00165##
[0558] is selected from H or Me;
R2 of
##STR00166##
[0559] is selected from alkyl or cycloalkyl;
X of
##STR00167##
[0560] is 1-2 substitutents independently selected from halogen,
hydroxy, methoxy, nitro and trifluoromethyl
Z of
##STR00168##
[0561] is O or NH;
HET of
##STR00169##
[0562] is mono- or fused bicyclic heteroaryl; and - - - of Formula
(LIX) is an optional double bond.
[0563] In a particular embodiment, the ILM of the compound has a
chemical structure as represented by:
##STR00170##
[0564] In a particular embodiment, the ILM of the compound has a
chemical structure selected from the group consisting of:
##STR00171## ##STR00172##
[0565] The term "independently" is used herein to indicate that the
variable, which is independently applied, varies independently from
application to application.
[0566] The term "alkyl" shall mean within its context a linear,
branch-chained or cyclic fully saturated hydrocarbon radical or
alkyl group, preferably a C.sub.1-C.sub.10, more preferably a
C.sub.1-C.sub.6, alternatively a C.sub.1-C.sub.3 alkyl group, which
may be optionally substituted. Examples of alkyl groups are methyl,
ethyl, n-butyl, sec-butyl, n-hexyl, n-heptyl, n-octyl, n-nonyl,
n-decyl, isopropyl, 2-methylpropyl, cyclopropyl, cyclopropylmethyl,
cyclobutyl, cyclopentyl, cyclopen-tylethyl, cyclohexylethyl and
cyclohexyl, among others. In certain embodiments, the alkyl group
is end-capped with a halogen group (At, Br, Cl, F, or I). In
certain preferred embodiments, compounds according to the present
disclosure which may be used to covalently bind to dehalogenase
enzymes. These compounds generally contain a side chain (often
linked through a polyethylene glycol group) which terminates in an
alkyl group which has a halogen substituent (often chlorine or
bromine) on its distal end which results in covalent binding of the
compound containing such a moiety to the protein.
[0567] The term "Alkenyl" refers to linear, branch-chained or
cyclic C.sub.2-C.sub.10 (preferably C.sub.2-C.sub.6) hydrocarbon
radicals containing at least one C.dbd.C bond.
[0568] The term "Alkynyl" refers to linear, branch-chained or
cyclic C.sub.2-C.sub.10 (preferably C.sub.2-C.sub.6) hydrocarbon
radicals containing at least one C.ident.C bond.
[0569] The term "alkylene" when used, refers to a
--(CH.sub.2).sub.n-- group (n is an integer generally from 0-6),
which may be optionally substituted. When substituted, the alkylene
group preferably is substituted on one or more of the methylene
groups with a C.sub.1-C.sub.6 alkyl group (including a cyclopropyl
group or a t-butyl group), but may also be substituted with one or
more halo groups, preferably from 1 to 3 halo groups or one or two
hydroxyl groups, O--(C.sub.1-C.sub.6 alkyl) groups or amino acid
sidechains as otherwise disclosed herein. In certain embodiments,
an alkylene group may be substituted with a urethane or alkoxy
group (or other group) which is further substituted with a
polyethylene glycol chain (of from 1 to 10, preferably 1 to 6,
often 1 to 4 ethylene glycol units) to which is substituted
(preferably, but not exclusively on the distal end of the
polyethylene glycol chain) an alkyl chain substituted with a single
halogen group, preferably a chlorine group. In still other
embodiments, the alkylene (often, a methylene) group, may be
substituted with an amino acid sidechain group such as a sidechain
group of a natural or unnatural amino acid, for example, alanine,
P-alanine, arginine, asparagine, aspartic acid, cysteine, cystine,
glutamic acid, glutamine, glycine, phenylalanine, histidine,
isoleucine, lysine, leucine, methionine, proline, serine,
threonine, valine, tryptophan or tyrosine.
[0570] The term "unsubstituted" shall mean substituted only with
hydrogen atoms. A range of carbon atoms which includes C.sub.0
means that carbon is absent and is replaced with H. Thus, a range
of carbon atoms which is C.sub.0-C.sub.6 includes carbons atoms of
1, 2, 3, 4, 5 and 6 and for Co, H stands in place of carbon.
[0571] The term "substituted" or "optionally substituted" shall
mean independently (i.e., where more than substituent occurs, each
substituent is independent of another substituent) one or more
substituents (independently up to five substitutents, preferably up
to three substituents, often 1 or 2 substituents on a moiety in a
compound according to the present disclosure and may include
substituents which themselves may be further substituted) at a
carbon (or nitrogen) position anywhere on a molecule within
context, and includes as substituents hydroxyl, thiol, carboxyl,
cyano (C.ident.N), nitro (NO.sub.2), halogen (preferably, 1, 2 or 3
halogens, especially on an alkyl, especially a methyl group such as
a trifluoromethyl), an alkyl group (preferably, C.sub.1-C.sub.10,
more preferably, C.sub.1-C.sub.6), aryl (especially phenyl and
substituted phenyl for example benzyl or benzoyl), alkoxy group
(preferably, C.sub.1-C.sub.6 alkyl or aryl, including phenyl and
substituted phenyl), thioether (C.sub.1-C.sub.6 alkyl or aryl),
acyl (preferably, C.sub.1-C.sub.6 acyl), ester or thioester
(preferably, C.sub.1-C.sub.6 alkyl or aryl) including alkylene
ester (such that attachment is on the alkylene group, rather than
at the ester function which is preferably substituted with a
C.sub.1-C.sub.6 alkyl or aryl group), preferably, C.sub.1-C.sub.6
alkyl or aryl, halogen (preferably, F or Cl), amine (including a
five- or six-membered cyclic alkylene amine, further including a
C.sub.1-C.sub.6 alkyl amine or a C.sub.1-C.sub.6 dialkyl amine
which alkyl groups may be substituted with one or two hydroxyl
groups) or an optionally substituted --N(C.sub.0-C.sub.6
alkyl)C(O)(O--C.sub.1-C.sub.6 alkyl) group (which may be optionally
substituted with a polyethylene glycol chain to which is further
bound an alkyl group containing a single halogen, preferably
chlorine substituent), hydrazine, amido, which is preferably
substituted with one or two C.sub.1-C.sub.6 alkyl groups (including
a carboxamide which is optionally substituted with one or two
C.sub.1-C.sub.6 alkyl groups), alkanol (preferably, C.sub.1-C.sub.6
alkyl or aryl), or alkanoic acid (preferably, C.sub.1-C.sub.6 alkyl
or aryl). Substituents according to the present disclosure may
include, for example --SiR.sub.1R.sub.2R.sub.3 groups where each of
R.sub.1 and R.sub.2 is as otherwise described herein and R.sub.3 is
H or a C.sub.1-C.sub.6 alkyl group, preferably R.sub.1, R.sub.2,
R.sub.3 in this context is a C.sub.1-C.sub.3 alkyl group (including
an isopropyl or t-butyl group). Each of the above-described groups
may be linked directly to the substituted moiety or alternatively,
the substituent may be linked to the substituted moiety (preferably
in the case of an aryl or heteraryl moiety) through an optionally
substituted --(CH.sub.2).sub.m or alternatively an optionally
substituted --(OCH.sub.2).sub.m--, --(OCH.sub.2CH.sub.2).sub.m-- or
--(CH.sub.2CH.sub.2O).sub.m-- group, which may be substituted with
any one or more of the above-described substituents. Alkylene
groups --(CH2).sub.m-- or --(CH.sub.2).sub.n-- groups or other
chains such as ethylene glycol chains, as identified above, may be
substituted anywhere on the chain. Preferred substitutents on
alkylene groups include halogen or C.sub.1-C.sub.6 (preferably
C.sub.1-C.sub.3) alkyl groups, which may be optionally substituted
with one or two hydroxyl groups, one or two ether groups
(O--C.sub.1-C.sub.6 groups), up to three halo groups (preferably
F), or a sideshain of an amino acid as otherwise described herein
and optionally substituted amide (preferably carboxamide
substituted as described above) or urethane groups (often with one
or two C.sub.0-C.sub.6 alkyl substitutents, which group(s) may be
further substituted). In certain embodiments, the alkylene group
(often a single methylene group) is substituted with one or two
optionally substituted C.sub.1-C.sub.6 alkyl groups, preferably
C.sub.1-C.sub.4 alkyl group, most often methyl or O-methyl groups
or a sidechain of an amino acid as otherwise described herein. In
the present disclosure, a moiety in a molecule may be optionally
substituted with up to five substituents, preferably up to three
substituents. Most often, in the present disclosure moieties which
are substituted are substituted with one or two substituents.
[0572] The term "substituted" (each substituent being independent
of any other substituent) shall also mean within its context of use
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, halogen, amido,
carboxamido, sulfone, including sulfonamide, keto, carboxy,
C.sub.1-C.sub.6 ester (oxyester or carbonylester), C.sub.1-C.sub.6
keto, urethane --O--C(O)--NR.sub.1R.sub.2 or
--N(R.sub.1)--C(O)--O--R.sub.1, nitro, cyano and amine (especially
including a C.sub.1-C.sub.6 alkylene-NR.sub.1R.sub.2, a mono- or
di-C.sub.1-C.sub.6 alkyl substituted amines which may be optionally
substituted with one or two hydroxyl groups). Each of these groups
contain unless otherwise indicated, within context, between 1 and 6
carbon atoms. In certain embodiments, preferred substituents will
include for example, --NH--, --NHC(O)--, --O--, .dbd.O,
--(CH.sub.2).sub.m-- (here, m and n are in context, 1, 2, 3, 4, 5
or 6), --S--, --S(O)--, SO.sub.2-- or --NH--C(O)--NH--,
--(CH.sub.2).sub.nOH, --(CH.sub.2).sub.nSH, --(CH.sub.2).sub.nCOOH,
C.sub.1-C.sub.6 alkyl, --(CH.sub.2).sub.nO--(C.sub.1-C.sub.6
alkyl), --(CH.sub.2).sub.nC(O)--(C.sub.1-C.sub.6 alkyl),
--(CH.sub.2).sub.nOC(O)--(C.sub.1-C.sub.6 alkyl),
--(CH.sub.2).sub.nC(O)O--(C.sub.1-C.sub.6 alkyl),
--(CH.sub.2).sub.nNHC(O)--R.sub.1,
--(CH.sub.2).sub.nC(O)--NR.sub.1R.sub.2, --(OCH.sub.2).sub.nOH,
--(CH.sub.2O).sub.nCOOH, C.sub.1-C.sub.6 alkyl,
--(OCH.sub.2).sub.nO--(C.sub.1-C.sub.6 alkyl),
--(CH.sub.2O).sub.nC(O)--(C.sub.1-C.sub.6 alkyl),
--(OCH.sub.2).sub.nNHC(O)--R.sub.1,
--(CH.sub.2O).sub.nC(O)--NR.sub.1R.sub.2, --S(O).sub.2--R.sub.S,
--S(O)--R.sub.S (R.sub.S is C.sub.1-C.sub.6 alkyl or a
--(CH.sub.2).sub.m--NR.sub.1R.sub.2 group), NO.sub.2, CN or halogen
(F, Cl, Br, I, preferably F or Cl), depending on the context of the
use of the substituent. R.sub.1 and R.sub.2 are each, within
context, H or a C.sub.1-C.sub.6 alkyl group (which may be
optionally substituted with one or two hydroxyl groups or up to
three halogen groups, preferably fluorine). The term "substituted"
shall also mean, within the chemical context of the compound
defined and substituent used, an optionally substituted aryl or
heteroaryl group or an optionally substituted heterocyclic group as
otherwise described herein. Alkylene groups may also be substituted
as otherwise disclosed herein, preferably with optionally
substituted C.sub.1-C.sub.6 alkyl groups (methyl, ethyl or
hydroxymethyl or hydroxyethyl is preferred, thus providing a chiral
center), a sidechain of an amino acid group as otherwise described
herein, an amido group as described hereinabove, or a urethane
group O--C(O)--NR.sub.1R.sub.2 group where R.sub.1 and R.sub.2 are
as otherwise described herein, although numerous other groups may
also be used as substituents. Various optionally substituted
moieties may be substituted with 3 or more substituents, preferably
no more than 3 substituents and preferably with 1 or 2
substituents. It is noted that in instances where, in a compound at
a particular position of the molecule substitution is required
(principally, because of valency), but no substitution is
indicated, then that substituent is construed or understood to be
H, unless the context of the substitution suggests otherwise.
[0573] The term "aryl" or "aromatic", in context, refers to a
substituted (as otherwise described herein) or unsubstituted
monovalent aromatic radical having a single ring (e.g., benzene,
phenyl, benzyl) or condensed rings (e.g., naphthyl, anthracenyl,
phenanthrenyl, etc.) and can be bound to the compound according to
the present disclosure at any available stable position on the
ring(s) or as otherwise indicated in the chemical structure
presented. Other examples of aryl groups, in context, may include
heterocyclic aromatic ring systems, "heteroaryl" groups having one
or more nitrogen, oxygen, or sulfur atoms in the ring (moncyclic)
such as imidazole, furyl, pyrrole, furanyl, thiene, thiazole,
pyridine, pyrimidine, pyrazine, triazole, oxazole or fused ring
systems such as indole, quinoline, indolizine, azaindolizine,
benzofurazan, etc., among others, which may be optionally
substituted as described above. Among the heteroaryl groups which
may be mentioned include nitrogen-containing heteroaryl groups such
as pyrrole, pyridine, pyridone, pyridazine, pyrimidine, pyrazine,
pyrazole, imidazole, triazole, triazine, tetrazole, indole,
isoindole, indolizine, azaindolizine, purine, indazole, quinoline,
dihydroquinoline, tetrahydroquinoline, isoquinoline,
dihydroisoquinoline, tetrahydroisoquinoline, quinolizine,
phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline,
pteridine, imidazopyridine, imidazotriazine, pyrazinopyridazine,
acridine, phenanthridine, carbazole, carbazoline, pyrimidine,
phenanthroline, phenacene, oxadiazole, benzimidazole,
pyrrolopyridine, pyrrolopyrimidine and pyridopyrimidine;
sulfur-containing aromatic heterocycles such as thiophene and
benzothiophene; oxygen-containing aromatic heterocycles such as
furan, pyran, cyclopentapyran, benzofuran and isobenzofuran; and
aromatic heterocycles comprising 2 or more hetero atoms selected
from among nitrogen, sulfur and oxygen, such as thiazole,
thiadizole, isothiazole, benzoxazole, benzothiazole,
benzothiadiazole, phenothiazine, isoxazole, furazan, phenoxazine,
pyrazoloxazole, imidazothiazole, thienofuran, furopyrrole,
pyridoxazine, furopyridine, furopyrimidine, thienopyrimidine and
oxazole, among others, all of which may be optionally
substituted.
[0574] The term "substituted aryl" refers to an aromatic
carbocyclic group comprised of at least one aromatic ring or of
multiple condensed rings at least one of which being aromatic,
wherein the ring(s) are substituted with one or more substituents.
For example, an aryl group can comprise a substituent(s) selected
from: --(CH.sub.2).sub.nOH,
--(CH.sub.2).sub.n--O--(C.sub.1-C.sub.6)alkyl,
--(CH.sub.2).sub.n--O--(CH.sub.2).sub.n--(C.sub.1-C.sub.6)alkyl,
--(CH.sub.2).sub.n--C(O)(C.sub.0-C.sub.6) alkyl,
--(CH.sub.2).sub.n--C(O)O(C.sub.0-C.sub.6)alkyl,
--(CH.sub.2).sub.n--OC(O)(C.sub.0-C.sub.6)alkyl, amine, mono- or
di-(C.sub.1-C.sub.6 alkyl) amine wherein the alkyl group on the
amine is optionally substituted with 1 or 2 hydroxyl groups or up
to three halo (preferably F, Cl) groups, OH, COOH, C.sub.1-C.sub.6
alkyl, preferably CH.sub.3, CF.sub.3, OMe, OCF.sub.3, NO.sub.2, or
CN group (each of which may be substituted in ortho-, meta- and/or
para-positions of the phenyl ring, preferably para-), an optionally
substituted phenyl group (the phenyl group itself is preferably
substituted with a linker group attached to a PTM group, including
a ULM group), and/or at least one of F, Cl, OH, COOH, CH.sub.3,
CF.sub.3, OMe, OCF.sub.3, NO.sub.2, or CN group (in ortho-, meta-
and/or para-positions of the phenyl ring, preferably para-), a
naphthyl group, which may be optionally substituted, an optionally
substituted heteroaryl, preferably an optionally substituted
isoxazole including a methylsubstituted isoxazole, an optionally
substituted oxazole including a methylsubstituted oxazole, an
optionally substituted thiazole including a methyl substituted
thiazole, an optionally substituted isothiazole including a methyl
substituted isothiazole, an optionally substituted pyrrole
including a methylsubstituted pyrrole, an optionally substituted
imidazole including a methylimidazole, an optionally substituted
benzimidazole or methoxybenzylimidazole, an optionally substituted
oximidazole or methyloximidazole, an optionally substituted diazole
group, including a methyldiazole group, an optionally substituted
triazole group, including a methylsubstituted triazole group, an
optionally substituted pyridine group, including a
halo-(preferably, F) or methylsubstituted pyridine group or an
oxapyridine group (where the pyridine group is linked to the phenyl
group by an oxygen), an optionally substituted furan, an optionally
substituted benzofuran, an optionally substituted
dihydrobenzofuran, an optionally substituted indole, indolizine or
azaindolizine (2, 3, or 4-azaindolizine), an optionally substituted
quinoline, and combinations thereof.
[0575] "Carboxyl" denotes the group --C(O)OR, where R is hydrogen,
alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl or
substituted heteroaryl, whereas these generic substituents have
meanings which are identical with definitions of the corresponding
groups defined herein.
[0576] The term "heteroaryl" or "hetaryl" can mean but is in no way
limited to an optionally substituted quinoline (which may be
attached to the pharmacophore or substituted on any carbon atom
within the quinoline ring), an optionally substituted indole
(including dihydroindole), an optionally substituted indolizine, an
optionally substituted azaindolizine (2, 3 or 4-azaindolizine) an
optionally substituted benzimidazole, benzodiazole, benzoxofuran,
an optionally substituted imidazole, an optionally substituted
isoxazole, an optionally substituted oxazole (preferably methyl
substituted), an optionally substituted diazole, an optionally
substituted triazole, a tetrazole, an optionally substituted
benzofuran, an optionally substituted thiophene, an optionally
substituted thiazole (preferably methyl and/or thiol substituted),
an optionally substituted isothiazole, an optionally substituted
triazole (preferably a 1,2,3-triazole substituted with a methyl
group, a triisopropylsilyl group, an optionally substituted
--(CH.sub.2).sub.m--O--C.sub.1-C.sub.6 alkyl group or an optionally
substituted --(CH.sub.2).sub.m--C(O)--O--C.sub.1-C.sub.6 alkyl
group), an optionally substituted pyridine (2-, 3, or 4-pyridine)
or a group according to the chemical structure:
##STR00173##
wherein: [0577] S.sup.c is CHR.sup.SS, NR.sup.URE, or O; R.sup.HET
is H, CN, NO.sub.2, halo (preferably Cl or F), optionally
substituted C.sub.1-C.sub.6 alkyl (preferably substituted with one
or two hydroxyl groups or up to three halo groups (e.g. CF.sub.3),
optionally substituted O(C.sub.1-C.sub.6 alkyl) (preferably
substituted with one or two hydroxyl groups or up to three halo
groups) or an optionally substituted acetylenic group
--C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6 alkyl
group (preferably C.sub.1-C.sub.3 alkyl); [0578] R.sup.SS is H, CN,
NO.sub.2, halo (preferably F or Cl), optionally substituted
C.sub.1-C.sub.6 alkyl (preferably substituted with one or two
hydroxyl groups or up to three halo groups), optionally substituted
O--(C.sub.1-C.sub.6 alkyl) (preferably substituted with one or two
hydroxyl groups or up to three halo groups) or an optionally
substituted --C(O)(C.sub.1-C.sub.6 alkyl) (preferably substituted
with one or two hydroxyl groups or up to three halo groups); [0579]
R.sup.URE is H, a C.sub.1-C.sub.6 alkyl (preferably H or
C.sub.1-C.sub.3 alkyl) or a --C(O)(C.sub.1-C.sub.6 alkyl), each of
which groups is optionally substituted with one or two hydroxyl
groups or up to three halogen, preferably fluorine groups, or an
optionally substituted heterocycle, for example piperidine,
morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene,
piperidine, piperazine, each of which is optionally substituted,
and [0580] Y.sup.C is N or C--R.sup.YC, where R.sup.YC is H, OH,
CN, NO.sub.2, halo (preferably Cl or F), optionally substituted
C.sub.1-C.sub.6 alkyl (preferably substituted with one or two
hydroxyl groups or up to three halo groups (e.g. CF.sub.3),
optionally substituted O(C.sub.1-C.sub.6 alkyl) (preferably
substituted with one or two hydroxyl groups or up to three halo
groups) or an optionally substituted acetylenic group
--C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6 alkyl
group (preferably C.sub.1-C.sub.3 alkyl).
[0581] The terms "aralkyl" and "heteroarylalkyl" refer to groups
that comprise both aryl or, respectively, heteroaryl as well as
alkyl and/or heteroalkyl and/or carbocyclic and/or heterocycloalkyl
ring systems according to the above definitions.
[0582] The term "arylalkyl" as used herein refers to an aryl group
as defined above appended to an alkyl group defined above. The
arylalkyl group is attached to the parent moiety through an alkyl
group wherein the alkyl group is one to six carbon atoms. The aryl
group in the arylalkyl group may be substituted as defined
above.
[0583] The term "Heterocycle" refers to a cyclic group which
contains at least one heteroatom, e.g., N, O or S, and may be
aromatic (heteroaryl) or non-aromatic. Thus, the heteroaryl
moieties are subsumed under the definition of heterocycle,
depending on the context of its use. Exemplary heteroaryl groups
are described hereinabove.
[0584] Exemplary heterocyclics include: azetidinyl, benzimidazolyl,
1,4-benzodioxanyl, 1,3-benzodioxolyl, benzoxazolyl, benzothiazolyl,
benzothienyl, dihydroimidazolyl, dihydropyranyl, dihydrofuranyl,
dioxanyl, dioxolanyl, ethyleneurea, 1,3-dioxolane, 1,3-dioxane,
1,4-dioxane, furyl, homopiperidinyl, imidazolyl, imidazolinyl,
imidazolidinyl, indolinyl, indolyl, isoquinolinyl,
isothiazolidinyl, isothiazolyl, isoxazolidinyl, isoxazolyl,
morpholinyl, naphthyridinyl, oxazolidinyl, oxazolyl, pyridone,
2-pyrrolidone, pyridine, piperazinyl, N-methylpiperazinyl,
piperidinyl, phthalimide, succinimide, pyrazinyl, pyrazolinyl,
pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, pyrrolyl,
quinolinyl, tetrahydrofuranyl, tetrahydropyranyl,
tetrahydroquinoline, thiazolidinyl, thiazolyl, thienyl,
tetrahydrothiophene, oxane, oxetanyl, oxathiolanyl, thiane among
others.
[0585] Heterocyclic groups can be optionally substituted with a
member selected from the group consisting of alkoxy, substituted
alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl,
substituted cycloalkenyl, acyl, acylamino, acyloxy, amino,
substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido,
cyano, halogen, hydroxyl, keto, thioketo, carboxy, carboxyalkyl,
thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol,
thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,
heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino,
alkoxyamino, nitro, --SO-alkyl, --SO-substituted alkyl, --SOaryl,
--SO-heteroaryl, --SO2-alkyl, --SO2-substituted alkyl, --SO2-aryl,
oxo (.dbd.O), and --SO2-heteroaryl. Such heterocyclic groups can
have a single ring or multiple condensed rings. Examples of
nitrogen heterocycles and heteroaryls include, but are not limited
to, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine,
pyridazine, indolizine, isoindole, indole, indazole, purine,
quinolizine, isoquinoline, quinoline, phthalazine,
naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine,
carbazole, carboline, phenanthridine, acridine, phenanthroline,
isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine,
imidazolidine, imidazoline, piperidine, piperazine, indoline,
morpholino, piperidinyl, tetrahydrofuranyl, and the like as well as
N-alkoxy-nitrogen containing heterocycles. The term "heterocyclic"
also includes bicyclic groups in which any of the heterocyclic
rings is fused to a benzene ring or a cyclohexane ring or another
heterocyclic ring (for example, indolyl, quinolyl, isoquinolyl,
tetrahydroquinolyl, and the like).
[0586] The term "cycloalkyl" can mean but is in no way limited to
univalent groups derived from monocyclic or polycyclic alkyl groups
or cycloalkanes, as defined herein, e.g., saturated monocyclic
hydrocarbon groups having from three to twenty carbon atoms in the
ring, including, but not limited to, cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl and the like. The term
"substituted cycloalkyl" can mean but is in no way limited to a
monocyclic or polycyclic alkyl group and being substituted by one
or more substituents, for example, amino, halogen, alkyl,
substituted alkyl, carbyloxy, carbylmercapto, aryl, nitro, mercapto
or sulfo, whereas these generic substituent groups have meanings
which are identical with definitions of the corresponding groups as
defined in this legend.
[0587] "Heterocycloalkyl" refers to a monocyclic or polycyclic
alkyl group in which at least one ring carbon atom of its cyclic
structure being replaced with a heteroatom selected from the group
consisting of N, O, S or P. "Substituted heterocycloalkyl" refers
to a monocyclic or polycyclic alkyl group in which at least one
ring carbon atom of its cyclic structure being replaced with a
heteroatom selected from the group consisting of N, O, S or P and
the group is containing one or more substituents selected from the
group consisting of halogen, alkyl, substituted alkyl, carbyloxy,
carbylmercapto, aryl, nitro, mercapto or sulfo, whereas these
generic substituent group have meanings which are identical with
definitions of the corresponding groups as defined in this
legend.
[0588] The term "hydrocarbyl" shall mean a compound which contains
carbon and hydrogen and which may be fully saturated, partially
unsaturated or aromatic and includes aryl groups, alkyl groups,
alkenyl groups and alkynyl groups.
[0589] The term "independently" is used herein to indicate that the
variable, which is independently applied, varies independently from
application to application.
[0590] The term "lower alkyl" refers to methyl, ethyl or
propyl.
[0591] The term "lower alkoxy" refers to methoxy, ethoxy or
propoxy.
[0592] In any of the embodiments described herein, the W, X, Y, Z,
G, G', R, R', R'', Q1-Q4, A, and Rn can independently be covalently
coupled to a linker and/or a linker to which is attached one or
more PTM, ULM, ILM or ILM' groups.
[0593] Exemplary MLMs
[0594] In certain additional embodiments, the MLM of the
bifunctional compound comprises chemical moieties such as
substituted imidazolines, substituted spiro-indolinones,
substituted pyrrolidines, substituted piperidinones, substituted
morpholinones, substituted pyrrolopyrimidines, substituted
imidazolopyridines, substituted thiazoloimidazoline, substituted
pyrrolopyrrolidinones, and substituted isoquinolinones.
[0595] In additional embodiments, the MLM comprises the core
structures mentioned above with adjacent bis-aryl substitutions
positioned as cis- or trans-configurations.
[0596] In still additional embodiments, the MLM comprises part of
structural features as in RG7112, RG7388, SAR405838, AMG-232,
AM-7209, DS-5272, MK-8242, and NVP-CGM-097, and analogs or
derivatives thereof.
[0597] In certain preferred embodiments, MLM is a derivative of
substituted imidazoline represented as Formula (A-1), or
thiazoloimidazoline represented as Formula (A-2), or spiro
indolinone represented as Formula (A-3), or pyrollidine represented
as Formula (A-4), or piperidinone/morphlinone represented as
Formula (A-5), or isoquinolinone represented as Formula (A-6), or
pyrollopyrimidine/imidazolopyridine represented as Formula (A-7),
or pyrrolopyrrolidinone/imidazolopyrrolidinone represented as
Formula (A-8).
##STR00174## ##STR00175##
wherein above Formula (A-1) through Formula (A-8), X of Formula
(A-1) through Formula (A-8) is selected from the group consisting
of carbon, oxygen, sulfur, sulfoxide, sulfone, and N--R.sup.a;
[0598] R.sup.a is independently H or an alkyl group with carbon
number 1 to 6; Y and Z of Formula (A-1) through Formula (A-8) are
independently carbon or nitrogen; A, A' and A'' of Formula (A-1)
through Formula (A-8) are independently selected from C, N, O or S,
can also be one or two atoms forming a fused bicyclic ring, or a
6,5- and 5,5-fused aromatic bicyclic group; R.sub.1, R.sub.2 of
Formula (A-1) through Formula (A-8) are independently selected from
the group consisting of an aryl or heteroaryl group, a heteroaryl
group having one or two heteroatoms independently selected from
sulfur or nitrogen, wherein the aryl or heteroaryl group can be
mono-cyclic or bi-cyclic, or unsubstituted or substituted with one
to three substituents independently selected from the group
consisting of: [0599] halogen, --CN, C1 to C6 alkyl group, C3 to C6
cycloalkyl, --OH, alkoxy with 1 to 6 carbons, fluorine substituted
alkoxy with 1 to 6 carbons, sulfoxide with 1 to 6 carbons, sulfone
with 1 to 6 carbons, ketone with 2 to 6 carbons, amides with 2 to 6
carbons, and dialkyl amine with 2 to 6 carbons; R.sub.3, R.sub.4 of
Formula (A-1) through Formula (A-8) are independently selected from
the group consisting of H, methyl and C1 to C6 alkyl; R.sub.5 of
Formula (A-1) through Formula (A-8) is selected from the group
consisting of an aryl or heteroaryl group, a heteroaryl group
having one or two heteroatoms independently selected from sulfur or
nitrogen, wherein the aryl or heteroaryl group can be mono-cyclic
or bi-cyclic, or unsubstituted or substituted with one to three
substituents independently selected from the group consisting of:
[0600] halogen, --CN, C1 to C6 alkyl group, C3 to C6 cycloalkyl,
--OH, alkoxy with 1 to 6 carbons, fluorine substituted alkoxy with
1 to 6 carbons, sulfoxide with 1 to 6 carbons, sulfone with 1 to 6
carbons, ketone with 2 to 6 carbons, amides with 2 to 6 carbons,
dialkyl amine with 2 to 6 carbons, alkyl ether (C2 to C6), alkyl
ketone (C3 to C6), morpholinyl, alkyl ester (C3 to C6), alkyl
cyanide (C3 to C6); R.sub.6 of Formula (A-1) through Formula (A-8)
is H or --C(.dbd.O)R.sup.b, wherein [0601] R.sup.b of Formula (A-1)
through Formula (A-8) is selected from the group consisting of
alkyl, cycloalkyl, mono-, di- or tri-substituted aryl or
heteroaryl, 4-morpholinyl, 1-(3-oxopiperazunyl), 1-piperidinyl,
4-N--R.sup.c-morpholinyl, 4-R.sup.c-1-piperidinyl, and
3-R.sup.c-1-piperidinyl, wherein [0602] R.sup.c of Formula (A-1)
through Formula (A-8) is selected from the group consisting of
alkyl, fluorine substituted alkyl, cyano alkyl,
hydroxyl-substituted alkyl, cycloalkyl, alkoxyalkyl, amide alkyl,
alkyl sulfone, alkyl sulfoxide, alkyl amide, aryl, heteroaryl,
mono-, bis- and tri-substituted aryl or heteroaryl, CH2CH2R.sup.d,
and CH2CH2CH2R.sup.d, wherein [0603] R.sup.d of Formula (A-1)
through Formula (A-8) is selected from the group consisting of
alkoxy, alkyl sulfone, alkyl sulfoxide, N-substituted carboxamide,
--NHC(O)-alkyl, --NH--SO.sub.2-alkyl, aryl, substituted aryl,
heteroaryl, substituted heteroaryl; R.sub.7 of Formula (A-1)
through Formula (A-8) is selected from the group consisting of H,
C1 to C6 alkyl, cyclic alkyl, fluorine substituted alkyl, cyano
substituted alkyl, 5- or 6-membered hetero aryl or aryl,
substituted 5- or 6-membered hetero aryl or aryl; R.sup.8 of
Formula (A-1) through Formula (A-8) is selected from the group
consisting of --R.sup.e--C(O)--R.sup.f, --R.sup.e-alkoxy,
--R.sup.e-aryl, --R.sup.e-heteroaryl, and
--R.sup.e--C(O)--R.sup.f--C(O)--R.sup.g, wherein: [0604] R.sup.e of
Formula (A-1) through Formula (A-8) is an alkylene with 1 to 6
carbons, or a bond; [0605] R.sup.f of Formula (A-1) through Formula
(A-8) is a substituted 4- to 7-membered heterocycle; [0606] R.sup.9
of Formula (A-1) through Formula (A-8) is selected from the group
consisting of aryl, hetero aryl, substituted aryl or heteroaryl,
and 4- to 7-membered heterocycle; R.sub.9 of Formula (A-1) through
Formula (A-8) is selected from the group consisting of a mono-,
bis- or tri-substituent on the fused bicyclic aromatic ring in
Formula (A-3), wherein the substitutents are independently selected
from the group consisting of halogen, alkene, alkyne, alkyl,
unsubstituted or substituted with Cl or F; R.sub.10 of Formula
(A-1) through Formula (A-8) is selected from the group consisting
of an aryl or heteroaryl group, wherein the heteroaryl group can
contain one or two heteroatoms as sulfur or nitrogen, aryl or
heteroaryl group can be mono-cyclic or bi-cyclic, the aryl or
heteroaryl group can be unsubstituted or substituted with one to
three substituents, including a halogen, F, Cl, --CN, alkene,
alkyne, C1 to C6 alkyl group, C1 to C6 cycloalkyl, --OH, alkoxy
with 1 to 6 carbons, fluorine substituted alkoxy with 1 to 6
carbons, sulfoxide with 1 to 6 carbons, sulfone with 1 to 6
carbons, ketone with 2 to 6 carbons; R.sub.11 of Formula (A-1)
through Formula (A-8) is --C(O)--N(R.sup.h)(R.sup.i), wherein
R.sup.h and R.sup.i are selected from groups consisting of the
following: [0607] H, C1 to C6 alkyl, alkoxy substituted alkyl,
sulfone substituted alkyl, aryl, heterol aryl, mono-, bis- or
tri-substituted aryl or hetero aryl, alkyl carboxylic acid,
heteroaryl carboxylic acid, alkyl carboxylic acid, fluorine
substituted alkyl carboxylic acid, aryl substituted cycloalkyl,
hetero aryl substituted cycloalkyl; wherein [0608] R.sup.h and
R.sup.i of Formula (A-1) through Formula (A-8) are independently
selected from the group consisting of H, connected to form a ring,
4-hydroxycyclohehexane; mono- and di-hydroxy substituted alkyl (C3
to C6); 3-hydroxycyclobutane; phenyl-4-carboxylic acid, and
substituted phenyl-4-carboxylic acid; R.sub.12 and R.sub.13 of
Formula (A-1) through Formula (A-8) are independently selected from
H, lower alkyl (C1 to C6), lower alkenyl (C2 to C6), lower alkynyl
(C2 to C6), cycloalkyl (4, 5 and 6-membered ring), substituted
cycloalkyl, cycloalkenyl, substituted cycloalkenyl, 5- and
6-membered aryl and heteroaryl, R12 and R13 can be connected to
form a 5- and 6-membered ring with or without substitution on the
ring; R.sub.14 of Formula (A-1) through Formula (A-8) is selected
from the group consisting of alkyl, substituted alkyl, alkenyl,
substituted alkenyl, aryl, substituted aryl, heteroaryl,
substituted heteroaryl, heterocycle, substituted heterocycle,
cycloalkyl, substituted cycloalkyl, cycloalkenyl and substituted
cycloalkenyl; R.sup.15 of Formula (A-1) through Formula (A-8) is
CN; R.sub.16 of Formula (A-1) through Formula (A-8) is selected
from the group consisting of C1-6 alkyl, C1-6 cycloalkyl, C2-6
alkenyl, C1-6 alkyl or C3-6 cycloalkyl with one or multiple
hydrogens replaced by fluorine, alkyl or cycloalkyl with one
CH.sub.2 replaced by S(.dbd.O), --S, or --S(.dbd.O).sub.2, alkyl or
cycloalkyl with terminal CH.sub.3 replaced by
S(.dbd.O).sub.2N(alkyl)(alkyl), --C(.dbd.O)N(alkyl)(alkyl),
--N(alkyl)S(.dbd.O).sub.2(alkyl), --C(.dbd.O)2(alkyl), --O(alkyl),
C1-6 alkyl or alkyl-cycloalkyl with hydron replaced by hydroxyl
group, a 3 to 7 membered cycloalkyl or heterocycloalkyl, optionally
containing a --(C=0)- group, or a 5 to 6 membered aryl or
heteroaryl group, which heterocycloalkyl or heteroaryl group can
contain from one to three heteroatoms independently selected from
O, N or S, and the cycloalkyl, heterocycloalkyl, aryl or heteroaryl
group can be unsubstituted or substituted with from one to three
substituents independently selected from halogen, C1-6 alkyl
groups, hydroxylated C1-6 alkyl, C1-6 alkyl containing thioether,
ether, sulfone, sulfoxide, fluorine substituted ether or cyano
group; R.sub.17 of Formula (A-1) through Formula (A-8) is selected
from the group consisting of (CH.sub.2)nC(O)NR.sup.kR.sup.l,
wherein R.sup.k and R.sup.l are independently selected from H, C1-6
alkyl, hydroxylated C1-6 alkyl, C1-6 alkoxy alkyl, C1-6 alkyl with
one or multiple hydrogens replaced by fluorine, C1-6 alkyl with one
carbon replaced by S(O), S(O)(O), C1-6 alkoxyalkyl with one or
multiple hydrogens replaced by fluorine, C.sub.1-6 alkyl with
hydrogen replaced by a cyano group, 5 and 6 membered aryl or
heteroaryl, alkyl aryl with alkyl group containing 1-6 carbons, and
alkyl heteroaryl with alkyl group containing 1-6 carbons, wherein
the aryl or heteroaryl group can be further substituted; R.sub.18
of Formula (A-1) through Formula (A-8) is selected from the group
consisting of substituted aryl, heteroaryl, alkyl, cycloalkyl, the
substitution is preferably --N(C1-4 alkyl)(cycloalkyl), --N(C1-4
alkyl)alkyl-cycloalkyl, and --N(C1-4
alkyl)[(alkyl)-(heterocycle-substituted)-cycloalkyl]; R.sub.19 of
Formula (A-1) through Formula (A-8) is selected from the group
consisting of aryl, heteroaryl, bicyclic heteroaryl, and these aryl
or heteroaryl groups can be substituted with halogen, C1-6 alkyl,
C1-6 cycloalkyl, CF.sub.3, F, CN, alkyne, alkyl sulfone, the
halogen substitution can be mon- bis- or tri-substituted; R.sub.20
and R.sub.21 of Formula (A-1) through Formula (A-8) are
independently selected from C1-6 alkyl, C1-6 cycloalkyl, C1-6
alkoxy, hydroxylated C1-6 alkoxy, and fluorine substituted C1-6
alkoxy, wherein R.sub.20 and R.sub.21 can further be connected to
form a 5, 6 and 7-membered cyclic or heterocyclic ring, which can
further be substituted; R.sub.22 of Formula (A-1) through Formula
(A-8) is selected from the group consisting of H, C1-6 alkyl, C1-6
cycloalkyl, carboxylic acid, carboxylic acid ester, amide, reverse
amide, sulfonamide, reverse sulfonamide, N-acyl urea,
nitrogen-containing 5-membered heterocycle, the 5-membered
heterocycles can be further substituted with C1-6 alkyl, alkoxy,
fluorine-substituted alkyl, CN, and alkylsulfone; R.sub.23 of
Formula (A-1) through Formula (A-8) is selected from aryl,
heteroaryl, --O-aryl, --O-heteroaryl, --O-alkyl,
--O-alkyl-cycloalkyl, --NH-alkyl, --NH-alkyl-cycloalkyl,
--N(H)-aryl, --N(H)-heteroaryl, --N(alkyl)-aryl,
--N(alkyl)-heteroaryl, the aryl or heteroaryl groups can be
substituted with halogen, C1-6 alkyl, hydroxylated C1-6 alkyl,
cycloalkyl, fluorine-substituted C1-6 alkyl, CN, alkoxy, alkyl
sulfone, amide and sulfonamide; R.sub.24 of Formula (A-1) through
Formula (A-8) is selected from the group consisting of --CH2-(C1-6
alkyl), --CH2-cycloalkyl, --CH2-aryl, CH2-heteroaryl, where alkyl,
cycloalkyl, aryl and heteroaryl can be substituted with halogen,
alkoxy, hydroxylated alkyl, cyano-substituted alkyl, cycloalkyl and
substituted cycloalkyl; R.sub.25 of Formula (A-1) through Formula
(A-8) is selected from the group consisting of C1-6 alkyl, C1-6
alkyl-cycloalkyl, alkoxy-substituted alkyl, hydroxylated alkyl,
aryl, heteroaryl, substituted aryl or heteroaryl, 5,6, and
7-membered nitrogen-containing saturated heterocycles, 5,6-fused
and 6,6-fused nitrogen-containing saturated heterocycles and these
saturated heterocycles can be substituted with C1-6 alkyl,
fluorine-substituted C1-6 alkyl, alkoxy, aryl and heteroaryl group;
R.sub.26 of Formula (A-1) through Formula (A-8) is selected from
the group consisting of C1-6 alkyl, C3-6 cycloalkyl, the alkyl or
cycloalkyl can be substituted with --OH, alkoxy,
fluorine-substituted alkoxy, fluorine-substituted alkyl,
--NH.sub.2, --NH-alkyl, NH--C(O)alkyl, --NH--S(O).sub.2-alkyl, and
--S(O).sub.2-alkyl; R.sub.27 of Formula (A-1) through Formula (A-8)
is selected from the group consisting of aryl, heteroaryl, bicyclic
heteroaryl, wherein the aryl or heteroaryl groups can be
substituted with C1-6 alkyl, alkoxy, NH2, NH-alkyl, halogen, or
--CN, and the substitution can be independently mono-, bis- and
tri-substitution; R.sub.28 of Formula (A-1) through Formula (A-8)
is selected from the group consisting of aryl, 5 and 6-membered
heteroaryl, bicyclic heteroaryl, cycloalkyl, saturated heterocycle
such as piperidine, piperidinone, tetrahydropyran,
N-acyl-piperidine, wherein the cycloalkyl, saturated heterocycle,
aryl or heteroaryl can be further substituted with --OH, alkoxy,
mono-, bis- or tri-substitution including halogen, --CN, alkyl
sulfone, and fluorine substituted alkyl groups; and R.sub.1'' of
Formula (A-1) through Formula (A-8) is selected from the group
consisting of alkyl, aryl substituted alkyl, alkoxy substituted
alkyl, cycloalkyl, aryl-substituted cycloalkyl, and alkoxy
substituted cycloalkyl.
[0609] In certain embodiments, the heterocycles in R.sup.f and
R.sup.g of Formula (A-1) through Formula (A-8) are substituted
pyrrolidine, substituted piperidine, substituted piperizine.
[0610] More specifically, non-limiting examples of MLMs include
those shown below as well as those `hybrid` molecules that arise
from the combination of 1 or more of the different features shown
in the molecules below.
[0611] Using MLM in Formula A-1 through A-8, the following PROTACs
can be prepared to target a particular protein for degradation,
where "L" is a connector (i.e. a linker group), and "PTM" is a
ligand binding to a target protein.
[0612] In certain embodiments, the description provides a
bifunctional molecule comprising a structure selected from the
group consisting of:
##STR00176## ##STR00177##
wherein X, R.sup.a, Y, Z, A, A', A'', R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sup.b, R.sup.c, R.sup.d, R.sub.7,
R.sup.e, R.sup.f, R.sup.g, R.sub.9, R.sub.10, R.sub.11, R.sub.12,
R.sub.13, R.sub.14, R.sub.15, R.sub.16, R.sub.17, R.sup.k, R.sup.l,
R.sub.18, R.sub.19, R.sub.20, R.sub.21, R.sub.22, R.sub.23,
R.sub.24, R.sub.25, R.sub.26, R.sub.27, R.sub.28, and R.sub.1'''
areas defined herein with regard to Formulas (A-1) through
(A-8).
[0613] In certain embodiments, the description provides
bifunctional or chimeric molecules with the structure: PTM-L-MLM,
wherein PTM is a protein target binding moiety coupled to an MLM by
L, wherein L is a bond (i.e., absent) or a chemical linker. In
certain embodiments, the MLM has a structure selected from the
group consisting of A-1-1, A-1-2, A-1-3, and A-1-4:
##STR00178##
wherein: R1' and R2' of Formulas A-1-1 through A-1-4 (i.e., A-1-1,
A-1-2, A-1-3, and A-1-4) are independently selected from the group
consisting of F, Cl, Br, I, acetylene, CN, CF.sub.3 and NO.sub.2;
R3' is selected from the group consisting of --OCH.sub.3,
--OCH.sub.2CH.sub.3, --OCH.sub.2CH.sub.2F,
--OCH.sub.2CH.sub.2OCH.sub.3, and --OCH(CH.sub.3).sub.2; R4' of
Formulas A-1-1 through A-1-4 is selected from the group consisting
of H, halogen, --CH.sub.3, --CF.sub.3, --OCH.sub.3,
--C(CH.sub.3).sub.3, --CH(CH.sub.3).sub.2, -cyclopropyl, --CN,
--C(CH.sub.3).sub.2OH, --C(CH.sub.3).sub.2OCH.sub.2CH.sub.3,
--C(CH.sub.3).sub.2CH.sub.2OH,
--C(CH.sub.3).sub.2CH.sub.2OCH.sub.2CH.sub.3,
--C(CH.sub.3).sub.2CH.sub.2OCH.sub.2CH.sub.2OH,
--C(CH.sub.3).sub.2CH.sub.2OCH.sub.2CH.sub.3,
--C(CH.sub.3).sub.2CN, --C(CH.sub.3).sub.2C(O)CH.sub.3,
--C(CH.sub.3).sub.2C(O)NHCH.sub.3,
--C(CH.sub.3).sub.2C(O)N(CH.sub.3).sub.2, --SCH.sub.3,
--SCH.sub.2CH.sub.3, --S(O).sub.2CH.sub.3,
--S(O.sub.2)CH.sub.2CH.sub.3, --NHC(CH.sub.3).sub.3,
--N(CH.sub.3).sub.2, pyrrolidinyl, and 4-morpholinyl; R5' of
Formulas A-1-1 through A-1-4 is selected from the group consisting
of halogen, -cyclopropyl, --S(O).sub.2CH.sub.3,
--S(O).sub.2CH.sub.2CH.sub.3, 1-pyrrolidinyl, --NH.sub.2,
--N(CH.sub.3).sub.2, and --NHC(CH.sub.3).sub.3; and R6' of Formulas
A-1-1 through A-1-4 is selected from the structures presented below
where the linker connection point is indicated as "*". Beside R6'
as the point for linker attachment, R4' can also serve as the
linker attachment position. In the case that R4' is the linker
connection site, linker will be connected to the terminal atom of
R4' groups shown above.
[0614] In certain embodiments, the linker connection position of
Formulas A-1-1 through A-1-4 is at least one of R4' or R6' or
both.
[0615] In certain embodiments, R6' of Formulas A-1-1 through A-1-4
is independently selected from the group consisting of H,
##STR00179## ##STR00180## ##STR00181##
and wherein indicates the point of attachment of the linker.
[0616] In certain embodiments, the linker of Formula A-4-1 through
A-4-6 is attached to at least one of R1', R2', R3', R4', R5', R6',
or a combination thereof.
[0617] In certain embodiments, the description provides
bifunctional or chimeric molecules with the structure: PTM-L-MLM,
wherein PTM is a protein target binding moiety coupled to an MLM by
L, wherein L is a bond (i.e., absent) or a chemical linker. In
certain embodiments, the MLM has a structure selected from the
group consisting of A-4-1, A-4-2, A-4-3, A-4-4, A-4-5, and
A-4-6:
##STR00182## ##STR00183##
wherein: R7' of Formula A-4-1 through A-4-6 (i.e., A-4-1, A-4-2,
A-4-3, A-4-4, A-4-5, and A-4-6) is a member selected from the group
consisting of halogen, mono-, and di- or tri-substituted halogen;
R8' of Formula A-4-1 through A-4-6 is selected from the group
consisting of H, --F, --Cl, --Br, --I, --CN, --NO.sub.2, ethylnyl,
cyclopropyl, methyl, ethyl, isopropyl, vinyl, methoxy, ethoxy,
isopropoxy, --OH, other C1-6 alkyl, other C1-6 alkenyl, and C1-6
alkynyl, mono-, di- or tri-substituted; R9' of Formula A-4-1
through A-4-6 is selected from the group consisting of alkyl,
substituted alkyl, alkenyl, substituted alkenyl, alkynyl,
substituted alkynyl, aryl, substituted aryl, hetero aryl,
substituted heteroaryl, cycloalkyl, substituted cycloalkyl,
alkenyl, and substituted cycloalkenyl; Z of Formula A-4-1 through
A-4-6 is selected from the group consisting of H, --OCH.sub.3,
--OCH.sub.2CH.sub.3, and halogen; R10' and R11' of Formula A-4-1
through A-4-6 are each independently selected from the group
consisting of H, (CH.sub.2).sub.n--R', (CH.sub.2).sub.n--NR'R'',
(CH.sub.2).sub.n--NR'COR'', (CH.sub.2).sub.n--NR'SO.sub.2R'',
(CH.sub.2).sub.n--COOH, (CH.sub.2).sub.n--COOR',
(CH).sub.n--CONR'R'', (CH.sub.2).sub.n--OR', (CH.sub.2).sub.n--SR',
(CH.sub.2).sub.n--SOR', (CH.sub.2).sub.n--CH(OH)--R',
(CH.sub.2).sub.n--COR', (CH.sub.2).sub.n--SO.sub.2R',
(CH.sub.2).sub.n--SONR'R'', (CH.sub.2).sub.n--SO.sub.2NR'R'',
(CH.sub.2CH.sub.2O).sub.m(CH.sub.2).sub.n--R',
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--OH,
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--OR',
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--NR'R'',
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--NR'COR'',
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--NR'SO.sub.2R'',
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--COOH,
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--COOR',
(CH.sub.2CH.sub.2O).sub.m--(CH2).sub.n--CONR'R'',
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--SO.sub.2R',
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--COR',
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--SONR'R'',
(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--SO.sub.2NR'R'',
(CH.sub.2).sub.p--(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.nR',
(CH.sub.2)p-(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--OH,
(CH.sub.2).sub.p--(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2)n-OR',
(CH.sub.2).sub.p--(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--NR'R'',
(CH.sub.2).sub.p--(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--NR'COR'',
(CH.sub.2).sub.p--(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--NR'SO.sub.-
2R'',
(CH.sub.2).sub.p--(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--COOH,
(CH.sub.2).sub.p--(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--COOR',
(CH.sub.2).sub.p--(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--CONR'R'',
(CH.sub.2)p-(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--SO.sub.2R',
(CH2).sub.p-(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--COR',
(CH.sub.2).sub.p--(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--SONR'R'',
(CH.sub.2).sub.p--(CH.sub.2CH.sub.2O).sub.m--(CH.sub.2).sub.n--SO.sub.2NR-
'R'', Aryl-(CH.sub.2).sub.n--COOH, and
heteroaryl-alkyl-CO-alkyl-NR'R''m, wherein the alkyl may be
substituted with OR', and heteroaryl-(CH.sub.2).sub.n-heterocycle
wherein the heterocycle may optionally be substituted with alkyl,
hydroxyl, COOR' and COR'; wherein R' and R'' are selected from H,
alkyl, alkyl substituted with halogen, hydroxyl, NH2, NH(alkyl),
N(alkyl)2, oxo, carboxy, cycloalkyl and heteroaryl; m, n, and p are
independently 0 to 6; R12' of Formula A-4-1 through A-4-6 is
selected from the group consisting of --O-(alkyl),
--O-(alkyl)-alkoxy, --C(O)-(alkyl), --C(OH)-alkyl-alkoxy,
--C(O)--NH-(alkyl), --C(O)--N-(alkyl).sub.2, --S(O)-(alkyl),
S(O).sub.2-(alkyl), --C(O)-(cyclic amine), and --O-aryl-(alkyl),
--O-aryl-(alkoxy); R1'' of Formula A-4-1 through A-4-6 is selected
from the group consisting of alkyl, aryl substituted alkyl, alkoxy
substituted alkyl, cycloalkyl, aryl-substituted cycloalkyl, and
alkoxy substituted cycloalkyl.
[0618] In any of the aspects or embodiments described herein, the
alkyl, alkoxy or the like can be a lower alkyl or lower alkoxy.
[0619] In certain embodiments, the linker connection position of
Formula A-4-1 through A-4-6 is at least one of Z, R8', R9', R10',
R11'', R12'', or R1''.
[0620] The method used to design chimeric molecules as presented in
A-1-1 through A-1-4, A-4-1 through A-4-6 can be applied to MLM with
formula A-2, A-3, A-5, A-6, A-7 and A-8, wherein the solvent
exposed area in the MLM can be connected to linker "L" which will
be attached to target protein ligand "PTM", to construct
PROTACs.
[0621] Exemplary MDM2 binding moieties include, but not limited,
the following:
[0622] the HDM2/MDM2 inhibitors identified in Vassilev, et al., In
vivo activation of the p53 pathway by small-molecule antagonists of
MDM2, SCIENCE vol:303, pag:844-848 (2004), and Schneekloth, et al.,
Targeted intracellular protein degradation induced by a small
molecule: En route to chemical proteomics, Bioorg. Med. Chem. Lett.
18 (2008) 5904-5908, including (or additionally) the compounds
nutlin-3, nutlin-2, and nutlin-1 (derivatized) as described below,
as well as all derivatives and analogs thereof:
##STR00184##
(derivatized where a linker group L or a -(L-MLM) group is
attached, for example, at the methoxy group or as a hydroxyl
group);
##STR00185##
(derivatized where a linker group L or a -(L-MLM) group is
attached, for example, at the methoxy group or hydroxyl group);
and
##STR00186##
(derivatized where a linker group L or a -(L-MLM) group is
attached, for example, via the methoxy group or as a hydroxyl
group).
[0623] Exemplary CLMs
[0624] Neo-Imide Compounds
[0625] In one aspect the description provides compounds useful for
binding and/or inhibiting cereblon. In certain embodiments, the
compound is selected from the group consisting of chemical
structures:
##STR00187##
wherein: [0626] W of Formulas (a) through (e) is independently
selected from the group CH.sub.2, CHR, C.dbd.O, SO.sub.2, NH, and
N-alkyl; [0627] X of Formulas (a) through (e) is independently
selected from the group O, S and H.sub.2; [0628] Y of Formulas (a)
through (e) is independently selected from the group CH.sub.2,
--C.dbd.CR', NH, N-alkyl, N-aryl, N-hetaryl, N-cycloalkyl,
N-heterocyclyl, O, and S; [0629] Z of Formulas (a) through (e) is
independently selected from the group O, and S or H.sub.2 except
that both X and Z cannot be H.sub.2; [0630] G and G' of Formulas
(a) through (e) are independently selected from the group H, alkyl
(linear, branched, optionally substituted), OH, R'OCOOR,
R'OCONRR'', CH.sub.2-heterocyclyl optionally substituted with R',
and benzyl optionally substituted with R'; [0631] Q1-Q4 of Formulas
(a) through (e) represent a carbon C substituted with a group
independently selected from R', N or N-oxide; [0632] A of Formulas
(a) through (e) is independently selected from the group H, alkyl
(linear, branched, optionally substituted), cycloalkyl, Cl and F;
[0633] R of Formulas (a) through (e) comprises, but is not limited
to: --CONR'R'', --OR', --NR'R'', --SR', --SO.sub.2R',
--SO.sub.2NR'R'', --CR'R''--, --CR'NR'R''--, (--CR'O).sub.nR'',
-aryl, -hetaryl, -alkyl (linear, branched, optionally substituted),
-cycloalkyl, -heterocyclyl, --P(O)(OR')R'', --P(O)R'R'',
--OP(O)(OR')R'', --OP(O)R'R'', --Cl, --F, --Br, --I, --CF.sub.3,
--CN, --NR'SO.sub.2NR'R'', --NR'CONR'R'', --CONR'COR'',
--NR'C(.dbd.N--CN)NR'R'', --C(.dbd.N--CN)NR'R'',
--NR'C(.dbd.N--CN)R'', --NR'C(.dbd.C--NO.sub.2)NR'R'',
--SO.sub.2NR'COR'', --NO.sub.2, --CO.sub.2R', --C(C.dbd.N--OR')R'',
--CR'.dbd.CR'R'', --CCR', --S(C.dbd.O)(C.dbd.N--R')R'', --SF.sub.5
and --OCF.sub.3 [0634] R' and R'' of Formulas (a) through (e) are
independently selected from a bond, H, alkyl, cycloalkyl, aryl,
heteroaryl, heterocyclic, --C(.dbd.O)R, heterocyclyl, each of which
is optionally substituted; [0635] n of Formulas (a) through (e) is
an integer from 1-10 (e.g., 1-4, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10);
[0636] of Formulas (a) through (e) represents a bond that may be
stereospecific ((R) or (S)) or on-stereospecific; and [0637]
R.sub.n of Formulas (a) through (e) comprises 1-4 independent
functional groups or atoms.
[0638] Exemplary CLMs
[0639] In any of the compounds described herein, the CLM comprises
a chemical structure selected from the group:
##STR00188##
wherein: [0640] W of Formulas (a) through (e) is independently
selected from the group CH.sub.2, CHR, C.dbd.O, SO.sub.2, NH, and
N-alkyl; [0641] X of Formulas (a) through (e) is independently
selected from the group O, S and H2; [0642] Y of Formulas (a)
through (e) is independently selected from the group CH.sub.2,
--C.dbd.CR', NH, N-alkyl, N-aryl, N-hetaryl, N-cycloalkyl,
N-heterocyclyl, O, and S; [0643] Z of Formulas (a) through (e) is
independently selected from the group O, and S or H2 except that
both X and Z cannot be H2; [0644] G and G' of Formulas (a) through
(e) are independently selected from the group H, alkyl (linear,
branched, optionally substituted), OH, R'OCOOR, R'OCONRR'',
CH2-heterocyclyl optionally substituted with R', and benzyl
optionally substituted with R'; [0645] Q1-Q4 of Formulas (a)
through (e) represent a carbon C substituted with a group
independently selected from R', N or N-oxide; [0646] A of Formulas
(a) through (e) is independently selected from the group H, alkyl
(linear, branched, optionally substituted), cycloalkyl, Cl and F;
[0647] R of Formulas (a) through (e) comprises, but is not limited
to: --CONR'R'', --OR', --NR'R'', --SR', --SO2R', --SO2NR'R'',
--CR'R''--, --CR'NR'R''--, -aryl, -hetaryl, -alkyl, -cycloalkyl,
-heterocyclyl, --P(O)(OR')R'', --P(O)R'R'', --OP(O)(OR')R'',
--OP(O)R'R'', --Cl, --F, --Br, --I, --CF.sub.3, --CN,
--NR'SO2NR'R'', --NR'CONR'R'', --CONR'COR'',
--NR'C(.dbd.N--CN)NR'R'', --C(.dbd.N--CN)NR'R'',
--NR'C(.dbd.N--CN)R'', --NR'C(.dbd.C--NO2)NR'R'', --SO2NR'COR'',
--NO2, --CO2R', --C(C.dbd.N--OR')R'', --CR'.dbd.CR'R'', --CCR',
--S(C.dbd.O)(C.dbd.N--R')R'', --SF5 and --OCF3 [0648] R' and R'' of
Formulas (a) through (e) are independently selected from a bond, H,
alkyl, cycloalkyl, aryl, heteroaryl, heterocyclic, --C(.dbd.O)R,
heterocyclyl, each of which is optionally substituted; [0649] n of
Formulas (a) through (e) is an integer from 1-10; [0650] of
Formulas (a) through (e) represents a bond that may be
stereospecific ((R) or (S)) or non-stereospecific; and [0651] Rn of
Formulas (a) through (e) comprises 1-4 independent functional
groups or atoms, for example, 0, N or S, and optionally, one of
which is modified to be covalently joined to a PTM, a chemical
linker group (L), a ULM, CLM (or CLM') or combination thereof.
[0652] In certain embodiments described herein, the CLM or ULM
comprises a chemical structure selected from the group:
##STR00189##
wherein:
[0653] W of Formula (g) is independently selected from the group
CH.sub.2, C.dbd.O, NH, and N-alkyl;
[0654] R of Formula (g) is independently selected from a H, methyl,
or optionally substituted alkyl (e.g., C.sub.1-C.sub.6 alkyl
(linear, branched, optionally substituted));
[0655] of Formula (g) represents a bond that may be stereospecific
((R) or (S)) or non-stereospecific; and
[0656] Rn of Formula (g) comprises 1-4 independently selected
functional groups or atoms, and optionally, one of which is
modified to be covalently joined to a PTM, a chemical linker group
(L), a ULM, CLM (or CLM') or combination thereof.
[0657] In any of the embodiments described herein, the W, X, Y, Z,
G, G', R, R', R'', Q1-Q4, A, and Rn of Formulas (a) through (g) can
independently be covalently coupled to a linker and/or a linker to
which is attached one or more PTM, ULM, CLM or CLM' groups.
[0658] In any of the aspects or embodiments described herein, Rn
comprises from 1 to 4 functional groups or atoms, for example, O,
OH, N, C1-C6 alkyl, C1-C6 alkoxy, amine, amide, or carboxy, and
optionally, one of which is modified to be covalently joined to a
PTM, a chemical linker group (L), a ULM, CLM (or CLM') or
combination thereof.
[0659] More specifically, non-limiting examples of CLMs include
those shown below as well as those "hybrid" molecules that arise
from the combination of 1 or more of the different features shown
in the molecules below.
##STR00190## ##STR00191## ##STR00192## ##STR00193## ##STR00194##
##STR00195## ##STR00196## ##STR00197## ##STR00198## ##STR00199##
##STR00200## ##STR00201## ##STR00202## ##STR00203## ##STR00204##
##STR00205## ##STR00206## ##STR00207## ##STR00208##
##STR00209##
[0660] In any of the compounds described herein, the CLM comprises
a chemical structure selected from the group:
##STR00210## ##STR00211## ##STR00212##
wherein: [0661] W of Formulas (h) through (ab) is independently
selected from CH.sub.2, CHR, C.dbd.O, SO.sub.2, NH, and N-alkyl;
[0662] Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4, Q.sub.5 of Formulas (h)
through (ab) are independently represent a carbon C substituted
with a group independently selected from R', N or N-oxide; [0663]
R.sup.1 of Formulas (h) through (ab) is selected from H, CN, C1-C3
alkyl; [0664] R.sup.2 of Formulas (h) through (ab) is selected from
the group H, CN, C1-C3 alkyl, CHF.sub.2, CF.sub.3, CHO; [0665]
R.sup.3 of Formulas (h) through (ab) is selected from H, alkyl,
substituted alkyl, alkoxy, substituted alkoxy; [0666] R.sup.4 of
Formulas (h) through (ab) is selected from H, alkyl, substituted
alkyl; [0667] R.sup.5 of Formulas (h) through (ab) is H or lower
alkyl; [0668] X of Formulas (h) through (ab) is C, CH or N; [0669]
R' of Formulas (h) through (ab) is selected from H, halogen, alkyl,
substituted alkyl, alkoxy, substituted alkoxy; [0670] R of Formulas
(h) through (ab) is H, OH, lower alkyl, lower alkoxy, cyano,
halogenated lower alkoxy, or halogenated lower alkyl [0671] of
Formulas (h) through (ab) is a single or double bond; and [0672]
the CLM is covalently joined to a PTM, a chemical linker group (L),
a ULM, CLM (or CLM') or combination thereof.
[0673] In any aspect or embodiment described herein, the CLM or
CLM' is covalently joined to a PTM, a chemical linker group (L), a
ULM, a CLM, a CLM', or a combination thereof via an R group (such
as, R, R.sup.1, R.sup.2, R.sup.3, R.sup.4 or R'), W, X, or a Q
group (such as, Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4, or Q.sub.5) of
Formulas (h) through (ab).
[0674] In any of the embodiments described herein, the CLM or CLM'
is covalently joined to a PTM, a chemical linker group (L), a ULM,
a CLM, a CLM', or a combination thereof via W, X, R, R.sup.1,
R.sup.2, R.sup.3, R.sup.4, R.sup.5, R', Q.sub.1, Q.sub.2, Q.sub.3,
Q.sub.4, and Q.sub.5 of Formulas (h) through (ab).
[0675] In any of the embodiments described herein, the W, X,
R.sup.1, R.sup.2, R.sup.3, R.sup.4, R', Q.sub.1, Q.sub.2, Q3, Q4,
and Q.sub.5 of Formulas (h) through (ab) can independently be
covalently coupled to a linker and/or a linker to which is attached
to one or more PTM, ULM, ULM', CLM or CLM' groups.
[0676] More specifically, non-limiting examples of CLMs include
those shown below as well as "hybrid" molecules or compounds that
arise from combining 1 or more features of the following
compounds:
##STR00213## ##STR00214##
wherein: [0677] W of Formulas (ac) through (an) is independently
selected from the group CH.sub.2, CHR, C.dbd.O, SO.sub.2, NH, and
N-alkyl; [0678] R.sup.1 of Formulas (ac) through (an) is selected
from the group H, CN, C1-C3 alkyl; [0679] R.sup.3 of Formulas (ac)
through (an) is selected from H, alkyl, substituted alkyl, alkoxy,
substituted alkoxy; [0680] R of Formulas (ac) through (an) is H;
[0681] is a single or double bond; and [0682] Rn of Formulas (ac)
through (an) comprises a functional group or an atom.
[0683] In any of the embodiments described herein, the W, R.sup.1,
R.sup.2, Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4, and Rn of Formulas
(ac) through (an) can independently be covalently coupled to a
linker and/or a linker to which is attached one or more PTM, ULM,
ULM', CLM or CLM' groups.
[0684] In any of the embodiments described herein, the R.sup.1,
R.sup.2, Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4, and Rn of Formulas
(ac) through (an) can independently be covalently coupled to a
linker and/or a linker to which is attached one or more PTM, ULM,
ULM', CLM or CLM' groups.
[0685] In any of the embodiments described herein, the Q.sub.1,
Q.sub.2, Q.sub.3, Q.sub.4, and Rn of Formulas (ac) through (an) can
independently be covalently coupled to a linker and/or a linker to
which is attached one or more PTM, ULM, ULM', CLM or CLM'
groups.
[0686] In any aspect or embodiment described herein, R. of Formulas
(ac) through (an) is modified to be covalently joined to the linker
group (L), a PTM, a ULM, a second CLM having the same chemical
structure as the CLM, a CLM', a second linker, or any multiple or
combination thereof.
[0687] In any aspect or embodiment described herein, the CLM is
selected from:
##STR00215## ##STR00216##
wherein R' is a halogen and R.sup.1 is as described above with
regard to Formulas (h) through (ab) or (ac) through (an).
[0688] In certain cases, the CLM can be imides that bind to
cereblon E3 ligase. These imides and linker attachment point can be
but not limited to the following structures:
##STR00217## ##STR00218##
wherein R' is a halogen.
[0689] Exemplary VLMs
[0690] In certain embodiments of the compounds as described herein,
ULM is VLM and comprises a chemical structure selected from the
group ULM-a:
##STR00219##
wherein:
[0691] a dashed line indicates the attachment of at least one PTM,
another ULM or VLM or MLM or ILM or CLM (i.e., ULM' or VLM' or CLM'
or ILM' or MLM'), or a chemical linker moiety coupling at least one
PTM, a ULM' or a VLM' or a CLM' or a ILM' or a MLM' to the other
end of the linker;
[0692] X.sup.1, X.sup.2 of Formula ULM-a are each independently
selected from the group of a bond, O, NR.sup.Y3, CR.sup.Y3R.sup.Y4,
C.dbd.O, C.dbd.S, SO, and SO.sub.2;
[0693] R.sup.Y3, R.sup.Y4 of Formula ULM-a are each independently
selected from the group of H, linear or branched C.sub.1-6 alkyl,
optionally substituted by 1 or more halo, optionally substituted
C.sub.1-6 alkoxyl (e.g., optionally substituted by 0-3 R.sup.P
groups);
[0694] R.sup.P of Formula ULM-a is 0, 1, 2, or 3 groups, each
independently selected from the group H, halo, --OH, C.sub.1-3
alkyl, C.dbd.O;
[0695] W.sup.3 of Formula ULM-a is selected from the group of an
optionally substituted -T-N(R.sup.1aR.sup.1b)X.sup.3, optionally
substituted -T-N(R.sup.1aR.sup.1b), optionally substituted -T-Aryl,
an optionally substituted -T-Heteroaryl, an optionally substituted
-T-Heterocycle, an optionally substituted --NR.sup.1-T-Aryl, an
optionally substituted --NR.sup.1-T-Heteroaryl or an optionally
substituted --NR.sup.1-T-Heterocycle;
[0696] X.sup.3 of Formula ULM-a is C.dbd.O, R.sup.1, R.sup.1a,
R.sup.1b;
[0697] each R.sup.1, R.sup.1a, R.sup.1b is independently selected
from the group consisting of H, linear or branched C.sub.1-C.sub.6
alkyl group optionally substituted by 1 or more halo or --OH
groups, R.sup.Y3C.dbd.O, R.sup.Y3C.dbd.S, R.sup.Y3SO,
R.sup.Y3SO.sub.2, N(R.sup.Y3R.sup.Y4)C.dbd.O,
N(R.sup.Y3R.sup.Y4)C.dbd.S, N(R.sup.Y3R.sup.Y4)SO, and
N(R.sup.Y3R.sup.Y4)SO.sub.2;
[0698] T of Formula ULM-a is covalently bonded to X.sup.1;
[0699] W.sup.4 of Formula ULM-a is an optionally substituted
--NR1-T-Aryl, an optionally substituted --NR1-T-Heteroaryl group or
an optionally substituted --NR1-T-Heterocycle, where --NR1 is
covalently bonded to X.sup.2 and R.sup.1 is H or CH3, preferably
H.
[0700] In any of the embodiments described herein, T is selected
from the group of an optionally substituted alkyl,
--(CH.sub.2).sub.n-- group, wherein each one of the methylene
groups is optionally substituted with one or two substituents
selected from the group of halogen, methyl, a linear or branched
C.sub.1-C.sub.6 alkyl group optionally substituted by 1 or more
halogen or --OH groups or an amino acid side chain optionally
substituted; and
[0701] n is 0 to 6, often 0, 1, 2, or 3, preferably 0 or 1.
[0702] In certain embodiments, W.sup.4 of Formula ULM-a is
##STR00220##
wherein
[0703] R.sub.14a, R.sub.14b, are each independently selected from
the group of H, haloalkyl, or optionally substituted alkyl.
[0704] In any of the embodiments, W.sup.5 of Formula ULM-a is
selected from the group of a phenyl or a 5-10 membered
heteroaryl,
[0705] R.sub.15 of Formula ULM-a is selected from the group of H,
halogen, CN, OH, NO.sub.2, N R.sub.14aR.sub.14b, OR.sub.14a,
CONR.sub.14aR.sub.14b, NR.sub.14aCOR.sub.14b,
SO.sub.2NR.sub.14aR.sub.14b, NR.sub.14a SO.sub.2R.sub.14b,
optionally substituted alkyl, optionally substituted haloalkyl,
optionally substituted haloalkoxy; optionally substituted aryl;
optionally substituted heteroaryl; optionally substituted
cycloalkyl; or optionally substituted cycloheteroalkyl;
[0706] In additional embodiments, W.sup.4 substituents for use in
the present disclosure also include specifically (and without
limitation to the specific compound disclosed) the W.sup.4
substituents which are found in the identified compounds disclosed
herein. Each of these W.sup.4 substituents may be used in
conjunction with any number of W.sup.3 substituents which are also
disclosed herein.
[0707] In certain additional embodiments, ULM-a, is optionally
substituted by 0-3 R.sup.P groups in the pyrrolidine moiety. Each
R.sup.P is independently H, halo, --OH, C1-3alkyl, C.dbd.O.
[0708] In any of the embodiments described herein, the W.sup.3,
W.sup.4 of Formula ULM-a can independently be covalently coupled to
a linker which is attached one or more PTM groups.
[0709] and wherein the dashed line indicates the site of attachment
of at least one PTM, another ULM (ULM') or a chemical linker moiety
coupling at least one PTM or a ULM' or both to ULM.
[0710] In certain embodiments, ULM is VHL and is represented by the
structure:
##STR00221##
wherein
[0711] W.sup.3 of Formula ULM-b is selected from the group of an
optionally substituted aryl, optionally substituted heteroaryl,
or
##STR00222##
[0712] R.sub.9 and R.sub.10 of Formula ULM-b are independently
hydrogen, optionally substituted alkyl, optionally substituted
cycloalkyl, optionally substituted hydroxyalkyl, optionally
substituted heteroaryl, or haloalkyl, or R.sub.9, R.sub.10, and the
carbon atom to which they are attached form an optionally
substituted cycloalkyl;
[0713] R.sub.11 of Formula ULM-b is selected from the group of an
optionally substituted heterocyclic, optionally substituted alkoxy,
optionally substituted heteroaryl, optionally substituted aryl,
##STR00223##
[0714] R.sub.12 of Formula ULM-b is selected from the group of H or
optionally substituted alkyl;
[0715] R.sub.13 of Formula ULM-b is selected from the group of H,
optionally substituted alkyl, optionally substituted alkylcarbonyl,
optionally substituted (cycloalkyl)alkylcarbonyl, optionally
substituted aralkylcarbonyl, optionally substituted arylcarbonyl,
optionally substituted (heterocyclyl)carbonyl, or optionally
substituted aralkyl;
[0716] R.sub.14a, R.sub.14b of Formula ULM-b, are each
independently selected from the group of H, haloalkyl, or
optionally substituted alkyl;
[0717] W.sup.5 of Formula ULM-b is selected from the group of a
phenyl or a 5-10 membered heteroaryl,
[0718] R.sub.15 of Formula ULM-b is selected from the group of H,
halogen, CN, OH, NO.sub.2, N R.sub.14aR.sub.14b, OR.sub.14a,
CONR.sub.14aR.sub.14b, NR.sub.14aCOR.sub.14b,
SO.sub.2NR.sub.14aR.sub.14b, NR.sub.14a SO.sub.2R.sub.14b,
optionally substituted alkyl, optionally substituted haloalkyl,
optionally substituted haloalkoxy; aryl, heteroaryl, cycloalkyl, or
cycloheteroalkyl (each optionally substituted);
[0719] R.sub.16 of Formula ULM-b is independently selected from the
group of H, halo, optionally substituted alkyl, optionally
substituted haloalkyl, hydroxy, or optionally substituted
haloalkoxy;
[0720] o of Formula ULM-b is 0, 1, 2, 3, or 4;
[0721] R.sub.18 of Formula ULM-b is independently selected from the
group of halo, optionally substituted alkoxy, cyano, optionally
substituted alkyl, haloalkyl, haloalkoxy or a linker; and
[0722] p of Formula ULM-b is 0, 1, 2, 3, or 4, and wherein the
dashed line indicates the site of attachment of at least one PTM,
another ULM (ULM') or a chemical linker moiety coupling at least
one PTM or a ULM' or both to ULM.
[0723] In certain embodiments, R.sub.15 of Formula ULM-b is
##STR00224##
wherein R.sub.17 is H, halo, optionally substituted
C.sub.3-6cycloalkyl, optionally substituted C.sub.1-6alkyl,
optionally substituted C.sub.1-6alkenyl, and C.sub.1-6haloalkyl;
and Xa is S or O.
[0724] In certain embodiments, R.sub.17 of Formula ULM-b is
selected from the group methyl, ethyl, isopropyl, and
cyclopropyl.
[0725] In certain additional embodiments, R.sub.15 of Formula ULM-b
is selected from the group consisting of:
##STR00225##
[0726] In certain embodiments, R.sub.11 of Formula ULM-b is
selected from the group consisting of:
##STR00226## ##STR00227##
[0727] In certain embodiments, ULM has a chemical structure
selected from the group of:
##STR00228##
wherein:
[0728] R.sub.1 of Formulas ULM-c, ULM-d, and ULM-e is H, ethyl,
isopropyl, tert-butyl, sec-butyl, cyclopropyl, cyclobutyl,
cyclopentyl, or cyclohexyl; optionally substituted alkyl,
optionally substituted hydroxyalkyl, optionally substituted
heteroaryl, or haloalkyl;
[0729] R.sub.14a of Formulas ULM-c, ULM-d, and ULM-e is H,
haloalkyl, optionally substituted alkyl, methyl, fluoromethyl,
hydroxymethyl, ethyl, isopropyl, or cyclopropyl;
[0730] R.sub.15 of Formulas ULM-c, ULM-d, and ULM-e is selected
from the group consisting of H, halogen, CN, OH, N02, optionally
substituted heteroaryl, optionally substituted aryl; optionally
substituted alkyl, optionally substituted haloalkyl, optionally
substituted haloalkoxy, cycloalkyl, or cycloheteroalkyl (each
optionally substituted);
[0731] X of Formulas ULM-c, ULM-d, and ULM-e is C, CH.sub.2, or
C.dbd.O
[0732] R.sub.13 of Formulas ULM-c, ULM-d, and ULM-e is absent or an
optionally substituted 5 or 6 membered heteroaryl; and
[0733] wherein the dashed line indicates the site of attachment of
at least one PTM, another ULM (ULM') or a chemical linker moiety
coupling at least one PTM or a ULM' or both to ULM.
[0734] In certain embodiments, ULM comprises a group according to
the chemical structure:
##STR00229##
wherein
[0735] R.sub.14a of Formula ULM-f is H, haloalkyl, optionally
substituted alkyl, methyl, fluoromethyl, hydroxymethyl, ethyl,
isopropyl, or cyclopropyl;
[0736] R.sub.9 of Formula ULM-f is H;
[0737] R.sub.10 of Formula ULM-f is H, ethyl, isopropyl,
tert-butyl, sec-butyl, cyclopropyl, cyclobutyl, cyclopentyl, or
cyclohexyl;
[0738] R.sub.11 of Formula ULM-f is
##STR00230##
[0739] or optionally substituted heteroaryl;
[0740] p of Formula ULM-f is 0, 1, 2, 3, or 4;
[0741] each R.sub.18 of Formula ULM-f is independently halo,
optionally substituted alkoxy, cyano, optionally substituted alkyl,
haloalkyl, haloalkoxy or a linker;
[0742] R.sub.12 of Formula ULM-f is H, C.dbd.O;
[0743] R.sub.13 of Formula ULM-f is H, optionally substituted
alkyl, optionally substituted alkylcarbonyl, optionally substituted
(cycloalkyl)alkylcarbonyl, optionally substituted aralkylcarbonyl,
optionally substituted arylcarbonyl, optionally substituted
(heterocyclyl)carbonyl, or optionally substituted aralkyl,
[0744] R.sub.15 of Formula ULM-f is selected from the group
consisting of H, halogen, Cl, CN, OH, NO.sub.2, optionally
substituted heteroaryl, optionally substituted aryl;
##STR00231##
and
[0745] wherein the dashed line of Formula ULM-f indicates the site
of attachment of at least one PTM, another ULM (ULM') or a chemical
linker moiety coupling at least one PTM or a ULM' or both to
ULM.
[0746] In certain embodiments, the ULM is selected from the
following structures:
##STR00232## ##STR00233## ##STR00234## ##STR00235##
where n is 0 or 1.
[0747] In certain embodiments, the ULM is selected from the
following structures:
##STR00236## ##STR00237## ##STR00238## ##STR00239## ##STR00240##
##STR00241## ##STR00242## ##STR00243## ##STR00244##
[0748] wherein, the phenyl ring in ULM-a1 through ULM-a15, ULM-b1
through ULM-b12, ULM-c1 through ULM-c15 and ULM-dl through ULM-d9
is optionally substituted with fluorine, lower alkyl and alkoxy
groups, and wherein the dashed line indicates the site of
attachment of at least one PTM, another ULM (ULM') or a chemical
linker moiety coupling at least one PTM or a ULM' or both to
ULM-a.
[0749] In one embodiment, the phenyl ring in ULM-a1 through
ULM-a15, ULM-b1 through ULM-b12, ULM-c1 through ULM-c15 and ULM-d1
through ULM-d9 can be functionalized as the ester to make it a part
of the prodrug.
[0750] In certain embodiments, the hydroxyl group on the
pyrrolidine ring of ULM-a1 through ULM-a15, ULM-b1 through ULM-b12,
ULM-c1 through ULM-c15 and ULM-d11 through ULM-d9, respectively,
comprises an ester-linked prodrug moiety.
[0751] In any of the aspects or embodiments described herein, the
ULM and where present, ULM', are each independently a group
according to the chemical structure:
##STR00245##
wherein: [0752] R.sup.1'' of ULM-g is an optionally substituted
C.sub.1-C.sub.6 alkyl group, an optionally substituted
--(CH.sub.2).sub.nOH, an optionally substituted
--(CH.sub.2).sub.nSH, an optionally substituted
(CH.sub.2).sub.n--O--(C.sub.1-C.sub.6)alkyl group, an optionally
substituted (CH.sub.2).sub.n--WCOCW--(C.sub.0-C.sub.6)alkyl group
containing an epoxide moiety WCOCW where each W is independently H
or a C.sub.1-C.sub.3 alkyl group, an optionally substituted
--(CH.sub.2).sub.nCOOH, an optionally substituted
--(CH.sub.2).sub.nC(O)--(C.sub.1-C.sub.6 alkyl), an optionally
substituted --(CH.sub.2).sub.nNHC(O)--R.sub.1, an optionally
substituted --(CH.sub.2).sub.nC(O)--NR.sub.1R.sub.2, an optionally
substituted --(CH.sub.2).sub.nOC(O)--NR.sub.1R.sub.2,
--(CH.sub.2O).sub.nH, an optionally substituted
--(CH.sub.2).sub.nOC(O)--(C.sub.1-C.sub.6 alkyl), an optionally
substituted --(CH.sub.2).sub.nC(O)--O--(C.sub.1-C.sub.6 alkyl), an
optionally substituted --(CH.sub.2O).sub.nCOOH, an optionally
substituted --(OCH.sub.2).sub.nO--(C.sub.1-C.sub.6 alkyl), an
optionally substituted --(CH.sub.2O).sub.nC(O)--(C.sub.1-C.sub.6
alkyl), an optionally substituted
--(OCH.sub.2).sub.nNHC(O)--R.sub.1, an optionally substituted
--(CH.sub.2O).sub.nC(O)--NR.sub.1R.sub.2,
--(CH.sub.2CH.sub.2O).sub.nH, an optionally substituted
--(CH.sub.2CH.sub.2O).sub.nCOOH, an optionally substituted
--(OCH.sub.2CH.sub.2).sub.nO--(C.sub.1-C.sub.6 alkyl), an
optionally substituted
--(CH.sub.2CH.sub.2O).sub.nC(O)--(C.sub.1-C.sub.6 alkyl), an
optionally substituted --(OCH.sub.2CH.sub.2).sub.nNHC(O)--R.sub.1,
an optionally substituted
--(CH.sub.2CH.sub.2O).sub.nC(O)--NR.sub.1R.sub.2, an optionally
substituted --SO.sub.2R.sub.S, an optionally substituted
S(O)R.sub.S, NO.sub.2, CN or halogen (F, Cl, Br, I, preferably F or
Cl); [0753] R.sub.1 and R.sub.2 of ULM-g are each independently H
or a C.sub.1-C.sub.6 alkyl group which may be optionally
substituted with one or two hydroxyl groups or up to three halogen
groups (preferably fluorine); [0754] R.sub.S of ULM-g is a
C.sub.1-C.sub.6 alkyl group, an optionally substituted aryl,
heteroaryl or heterocycle group or a
--(CH.sub.2).sub.mNR.sub.1R.sub.2 group; [0755] X and X' of ULM-g
are each independently C.dbd.O, C.dbd.S, --S(O), S(O).sub.2,
(preferably X and X' are both C.dbd.O); [0756] R.sup.2' of ULM-g is
an optionally substituted
--(CH.sub.2).sub.n--(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.walkyl
group, an optionally substituted
--(CH2).sub.n-(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.wNR.sub.1NR.s-
ub.2N group, an optionally substituted
--(CH.sub.2).sub.n--(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w-Aryl,
an optionally substituted
--(CH.sub.2).sub.n--(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w-Heter-
oaryl, an optionally substituted
--(CH.sub.2).sub.n--(C.dbd.O).sub.vNR.sub.1(SO.sub.2).sub.w-Heterocycle,
an optionally substituted
--NR.sup.1--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--
alkyl, an optionally substituted
--NR.sup.1--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--
-NR.sub.1NR.sub.2N, an optionally substituted
--NR.sup.1--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--
-NR.sub.1C(O)R.sub.1N, an optionally substituted
--NR.sup.1--(CH.sub.2).sub.n--(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).s-
ub.w-Aryl, an optionally substituted
--NR.sup.1--(CH.sub.2).sub.n--(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).s-
ub.w-Heteroaryl or an optionally substituted
--NR.sub.1--(CH.sub.2).sub.n--(C.dbd.O).sub.vNR.sub.1(SO.sub.2).sub.w-Het-
erocycle, an optionally substituted --X.sup.R2'-alkyl group; an
optionally substituted --X.sup.R2'-Aryl group; an optionally
substituted --X.sup.R2'-Heteroaryl group; an optionally substituted
--X.sup.R2'-Heterocycle group; an optionally substituted; [0757]
R.sup.3' of ULM-g is an optionally substituted alkyl, an optionally
substituted
--(CH.sub.2).sub.n--(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w-alkyl,
an optionally substituted
--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--NR.sub.1N-
R.sub.2N, an optionally substituted
--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--NR.sub.1C-
(O)R.sub.1N, an optionally substituted
--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--C(O)NR.su-
b.1R.sub.2, an optionally substituted
--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w-Aryl,
an optionally substituted
--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w-Heteroaryl-
, an optionally substituted
--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w-Heterocycl-
e, an optionally substituted
--NR.sup.1--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--
alkyl, an optionally substituted
--NR.sub.1--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--
-NR.sub.1NR.sub.2N, an optionally substituted
--NR.sup.1--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--
-NR.sub.1C(O)R.sub.1N, an optionally substituted
--NR.sup.1--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--
Aryl, an optionally substituted
--NR.sup.1--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--
Heteroaryl, an optionally substituted
--NR.sup.1--(CH.sub.2).sub.n--C(O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--
Heterocycle, an optionally substituted
--O--(CH.sub.2).sub.n--(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w-al-
kyl, an optionally substituted
--O--(CH.sub.2).sub.n--(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--N-
R.sub.1NR.sub.2N, an optionally substituted
--O--(CH.sub.2)n-(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w--NR.sub.-
1C(O)R.sub.1N, an optionally substituted
--O--(CH.sub.2)n-(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w-Aryl,
an optionally substituted
--O--(CH.sub.2).sub.n--(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w-He-
teroaryl or an optionally substituted
--O--(CH.sub.2).sub.n--(C.dbd.O).sub.u(NR.sub.1).sub.v(SO.sub.2).sub.w-He-
terocycle;
--(CH.sub.2).sub.n--(V).sub.n'--(CH.sub.2).sub.n--(V).sub.n'-al-
kyl group, an optionally substituted
--(CH.sub.2).sub.n--(V).sub.n'--(CH.sub.2).sub.n'--(V).sub.n'-Aryl
group, an optionally substituted
--(CH.sub.2).sub.n--(V).sub.n'--(CH.sub.2).sub.n--(V).sub.n'-Heteroaryl
group, an optionally substituted
--(CH.sub.2).sub.n--(V).sub.n'--(CH.sub.2).sub.n--(V).sub.n'-Heterocycle
group, an optionally substituted
--(CH.sub.2).sub.n--N(R.sub.1')(C.dbd.O).sub.m'--(V).sub.n'-alkyl
group, an optionally substituted
--(CH.sub.2).sub.n--N(R.sub.1')(C.dbd.O).sub.m'--(V).sub.n'-Aryl
group, an optionally substituted
--(CH.sub.2).sub.n--N(R.sub.1')(C.dbd.O).sub.m'--(V).sub.n'-Heteroaryl
group, an optionally substituted
--(CH.sub.2).sub.n--N(R.sub.1')(C.dbd.O).sub.m'--(V).sub.n'-Heterocycle
group, an optionally substituted --X.sup.R3'-alkyl group; an
optionally substituted --X.sup.R3'-Aryl group; an optionally
substituted --X.sup.R3'-Heteroaryl group; an optionally substituted
--X.sup.R3'-Heterocycle group; an optionally substituted; [0758]
R.sub.1N and R.sub.2N of ULM-g are each independently H,
C.sub.1-C.sub.6 alkyl which is optionally substituted with one or
two hydroxyl groups and up to three halogen groups or an optionally
substituted --(CH2).sub.n-Aryl, --(CH2).sub.n-Heteroaryl or
--(CH2).sub.n-Heterocycle group; [0759] V of ULM-g is O, S or
NR.sub.1; [0760] R.sub.1 of ULM-g is the same as above; [0761]
R.sup.1 and R.sub.1' of ULM-g are each independently H or a
C.sub.1-C.sub.3 alkyl group; [0762] X.sup.R2' and X.sup.R3' of
ULM-g are each independently an optionally substituted
--CH.sub.2).sub.n--,
--CH.sub.2).sub.n--CH(X.sub.v).dbd.CH(X.sub.v)-- (cis or trans),
--CH.sub.2).sub.n--CH.ident.CH--, --(CH.sub.2CH.sub.2O).sub.n-- or
a C.sub.3-C.sub.6 cycloalkyl group, where X.sub.v is H, a halo or a
C.sub.1-C.sub.3 alkyl group which is optionally substituted; [0763]
each m of ULM-g is independently 0, 1, 2, 3, 4, 5, 6; [0764] each
m' of ULM-g is independently 0 or 1; [0765] each n of ULM-g is
independently 0, 1, 2, 3, 4, 5, 6; [0766] each n' of ULM-g is
independently 0 or 1; [0767] each u of ULM-g is independently 0 or
1; [0768] each v of ULM-g is independently 0 or 1; [0769] each w of
ULM-g is independently 0 or 1; and [0770] any one or more of
R.sup.1', R.sup.2', R.sup.3', X and X' of ULM-g is optionally
modified to be covalently bonded to the PTM group through a linker
group when PTM is not ULM', or when PTM is ULM', any one or more of
R.sup.1', R.sup.2', R.sup.3', X and X' of each of ULM and ULM' are
optionally modified to be covalently bonded to each other directly
or through a linker group, or a pharmaceutically acceptable salt,
stereoisomer, solvate or polymorph thereof.
[0771] In any of the aspects or embodiments described herein, the
ULM and when present, ULM', are each independently a group
according to the chemical structure:
##STR00246##
wherein: [0772] each of R.sup.1', R.sup.2' and R.sup.3' of ULM-h
are the same as above and X is C.dbd.O, C.dbd.S, --S(O) group or a
S(O).sub.2 group, more preferably a C.dbd.O group, and [0773] any
one or more of R.sup.1', R.sup.2' and R.sup.3' of ULM-h are
optionally modified to bind a linker group to which is further
covalently bonded to the PTM group when PTM is not ULM', or when
PTM is ULM', any one or more of R.sup.1', R.sup.2', R.sup.3' of
each of ULM and ULM' are optionally modified to be covalently
bonded to each other directly or through a linker group, or [0774]
a pharmaceutically acceptable salt, enantiomer, diastereomer,
solvate or polymorph thereof.
[0775] In any of the aspects or embodiments described herein, the
ULM, and when present, ULM', are each independently according to
the chemical structure:
##STR00247##
wherein: [0776] any one or more of R.sup.1', R.sup.2' and R.sup.3'
of ULM-I are optionally modified to bind a linker group to which is
further covalently bonded to the PTM group when PTM is not ULM', or
when PTM is ULM', any one or more of R.sup.1', R.sup.2', R.sup.3'
of each of ULM and ULM' are optionally modified to be covalently
bonded to each other directly or through a linker group, or [0777]
a pharmaceutically acceptable salt, enantiomer, diastereomer,
solvate or polymorph thereof.
[0778] In further preferred aspects of the disclosure, R.sup.1' of
ULM-g through ULM-i is preferably a hydroxyl group or a group which
may be metabolized to a hydroxyl or carboxylic group, such that the
compound represents a prodrug form of an active compound. Exemplary
preferred R.sup.1' groups include, for example,
--(CH.sub.2).sub.nOH, (CH.sub.2).sub.n--O--(C.sub.1-C.sub.6)alkyl
group, --(CH.sub.2).sub.nCOOH, --(CH.sub.2O).sub.nH, an optionally
substituted --(CH.sub.2).sub.nOC(O)--(C.sub.1-C.sub.6 alkyl), or an
optionally substituted --(CH.sub.2).sub.nC(O)--O--(C.sub.1-C.sub.6
alkyl), wherein n is 0 or 1. Where R.sup.1' is or contains a
carboxylic acid group, a hydroxyl group or an amine group, the
hydroxyl group, carboxylic acid group or amine (each of which may
be optionally substituted), may be further chemically modified to
provide a covalent link to a linker group to which the PTM group
(including a ULM' group) is bonded;
[0779] X and X', where present, of ULM-g and ULM-h are preferably a
C.dbd.O, C.dbd.S, --S(O) group or a S(O).sub.2 group, more
preferably a C.dbd.O group;
[0780] R.sup.2' of ULM-g through ULM-i is preferably an optionally
substituted --NR.sup.1-T-Aryl, an optionally substituted
--NR.sup.1-T-Heteroaryl group or an optionally substituted
--NR.sup.1-T-Heterocycle, where R.sup.1 is H or CH.sub.3,
preferably H and T is an optionally substituted
--(CH.sub.2).sub.n-- group, wherein each one of the methylene
groups may be optionally substituted with one or two substituents,
preferably selected from halogen, an amino acid sidechain as
otherwise described herein or a C.sub.1-C.sub.3 alkyl group,
preferably one or two methyl groups, which may be optionally
substituted; and n is 0 to 6, often 0, 1, 2 or 3, preferably 0 or
1. Alternatively, T may also be a --(CH.sub.2O).sub.n-- group, a
--(OCH.sub.2).sub.n-- group, a --(CH.sub.2CH.sub.2O).sub.n-- group,
a --(OCH.sub.2CH.sub.2).sub.n-- group, all of which groups are
optionally substituted.
[0781] Preferred Aryl groups for R.sup.2' of ULM-g through ULM-i
include optionally substituted phenyl or naphthyl groups,
preferably phenyl groups, wherein the phenyl or naphthyl group is
connected to a PTM (including a ULM' group) with a linker group
and/or optionally substituted with a halogen (preferably F or Cl),
an amine, monoalkyl- or dialkyl amine (preferably, dimethylamine),
F, Cl, OH, COOH, C.sub.1-C.sub.6 alkyl, preferably CH.sub.3,
CF.sub.3, OMe, OCF.sub.3, NO.sub.2, or CN group (each of which may
be substituted in ortho-, meta- and/or para-positions of the phenyl
ring, preferably para-), an optionally substituted phenyl group
(the phenyl group itself is optionally connected to a PTM group,
including a ULM', with a linker group), and/or optionally
substituted with at least one of F, Cl, OH, COOH, CH.sub.3,
CF.sub.3, OMe, OCF.sub.3, NO.sub.2, or CN group (in ortho-, meta-
and/or para-positions of the phenyl ring, preferably para-), a
naphthyl group, which may be optionally substituted, an optionally
substituted heteroaryl, preferably an optionally substituted
isoxazole including a methylsubstituted isoxazole, an optionally
substituted oxazole including a methylsubstituted oxazole, an
optionally substituted thiazole including a methyl substituted
thiazole, an optionally substituted isothiazole including a methyl
substituted isothiazole, an optionally substituted pyrrole
including a methylsubstituted pyrrole, an optionally substituted
imidazole including a methylimidazole, an optionally substituted
benzimidazole or methoxybenzylimidazole, an optionally substituted
oximidazole or methyloximidazole, an optionally substituted diazole
group, including a methyldiazole group, an optionally substituted
triazole group, including a methylsubstituted triazole group, an
optionally substituted pyridine group, including a halo-
(preferably, F) or methyl substituted pyridine group or an
oxapyridine group (where the pyridine group is linked to the phenyl
group by an oxygen), an optionally substituted furan, an optionally
substituted benzofuran, an optionally substituted
dihydrobenzofuran, an optionally substituted indole, indolizine or
azaindolizine (2, 3, or 4-azaindolizine), an optionally substituted
quinoline, an optionally substituted group according to the
chemical structure:
##STR00248##
wherein: [0782] S.sup.c of ULM-g through ULM-i is CHR.sup.SS,
NR.sup.URE, or O; [0783] R.sup.HET of ULM-g through ULM-i is H, CN,
NO.sub.2, halo (preferably Cl or F), optionally substituted
C.sub.1-C.sub.6 alkyl (preferably substituted with one or two
hydroxyl groups or up to three halo groups (e.g. CF.sub.3),
optionally substituted O(C.sub.1-C.sub.6 alkyl) (preferably
substituted with one or two hydroxyl groups or up to three halo
groups) or an optionally substituted acetylenic group
--C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6 alkyl
group (preferably C.sub.1-C.sub.3 alkyl); [0784] R.sup.SS of ULM-g
through ULM-i is H, CN, NO.sub.2, halo (preferably F or Cl),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups), optionally substituted O--(C.sub.1-C.sub.6 alkyl)
(preferably substituted with one or two hydroxyl groups or up to
three halo groups) or an optionally substituted
--C(O)(C.sub.1-C.sub.6alkyl) (preferably substituted with one or
two hydroxyl groups or up to three halo groups); [0785] R.sup.URE
of ULM-g through ULM-i is H, a C.sub.1-C.sub.6 alkyl (preferably H
or C.sub.1-C.sub.3 alkyl) or a --C(O)(C.sub.1-C.sub.6 alkyl) each
of which groups is optionally substituted with one or two hydroxyl
groups or up to three halogen, preferably fluorine groups, or an
optionally substituted phenyl group, an optionally substituted
heteroaryl, or an optionally substituted heterocycle, preferably
for example piperidine, morpholine, pyrrolidine, tetrahydrofuran);
[0786] R.sup.PRO of ULM-g through ULM-i is H, optionally
substituted C.sub.1-C.sub.6 alkyl or an optionally substituted aryl
(phenyl or napthyl), heteroaryl or heterocyclic group selected from
the group consisting of oxazole, isoxazole, thiazole, isothiazole,
imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan,
dihydrofuran, tetrahydrofuran, thiene, dihydrothiene,
tetrahydrothiene, pyridine, piperidine, piperazine, morpholine,
quinoline, (each preferably substituted with a C.sub.1-C.sub.3
alkyl group, preferably methyl or a halo group, preferably F or
Cl), benzofuran, indole, indolizine, azaindolizine; [0787]
R.sup.PRO1 and R.sup.PRO2 of ULM-g through ULM-i are each
independently H, an optionally substituted C.sub.1-C.sub.3 alkyl
group or together form a keto group; and [0788] each n of ULM-g
through ULM-i is independently 0, 1, 2, 3, 4, 5, or 6 (preferably 0
or 1), or an optionally substituted heterocycle, preferably
tetrahydrofuran, tetrahydrothiene, piperidine, piperazine or
morpholine (each of which groups when substituted, are preferably
substituted with a methyl or halo (F, Br, Cl), each of which groups
may be optionally attached to a PTM group (including a ULM' group)
via a linker group.
[0789] In certain preferred aspects,
##STR00249##
of ULM-g through ULM-i is a
##STR00250##
group, where R.sup.PRO and n of ULM-g through ULM-i are the same as
above.
[0790] Preferred heteroaryl groups for R.sup.2' of ULM-g through
ULM-i include an optionally substituted quinoline (which may be
attached to the pharmacophore or substituted on any carbon atom
within the quinoline ring), an optionally substituted indole, an
optionally substituted indolizine, an optionally substituted
azaindolizine, an optionally substituted benzofuran, including an
optionally substituted benzofuran, an optionally substituted
isoxazole, an optionally substituted thiazole, an optionally
substituted isothiazole, an optionally substituted thiophene, an
optionally substituted pyridine (2-, 3, or 4-pyridine), an
optionally substituted imidazole, an optionally substituted
pyrrole, an optionally substituted diazole, an optionally
substituted triazole, a tetrazole, an optionally substituted
oximidazole, or a group according to the chemical structure:
##STR00251##
wherein: [0791] S.sup.c of ULM-g through ULM-i is CHR.sup.SS,
NR.sup.URE, or O; [0792] R.sup.HET of ULM-g through ULM-i is H, CN,
NO.sub.2, halo (preferably Cl or F), optionally substituted
C.sub.1-C.sub.6 alkyl (preferably substituted with one or two
hydroxyl groups or up to three halo groups (e.g. CF.sub.3),
optionally substituted O(C.sub.1-C.sub.6 alkyl) (preferably
substituted with one or two hydroxyl groups or up to three halo
groups) or an optionally substituted acetylenic group
--C.ident.C--R.sub.a where R.sub.a of ULM-g through ULM-i is H or a
C.sub.1-C.sub.6 alkyl group (preferably C.sub.1-C.sub.3 alkyl);
[0793] R.sup.SS of ULM-g through ULM-i is H, CN, NO.sub.2, halo
(preferably F or Cl), optionally substituted C.sub.1-C.sub.6 alkyl
(preferably substituted with one or two hydroxyl groups or up to
three halo groups), optionally substituted O--(C.sub.1-C.sub.6
alkyl) (preferably substituted with one or two hydroxyl groups or
up to three halo groups) or an optionally substituted
--C(O)(C.sub.1-C.sub.6alkyl) (preferably substituted with one or
two hydroxyl groups or up to three halo groups); [0794] R.sup.URE
of ULM-g through ULM-i is H, a C.sub.1-C.sub.6 alkyl (preferably H
or C.sub.1-C.sub.3 alkyl) or a --C(O)(C.sub.1-C.sub.6 alkyl), each
of which groups is optionally substituted with one or two hydroxyl
groups or up to three halogen, preferably fluorine groups, or an
optionally substituted heterocycle, for example piperidine,
morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene,
piperidine, piperazine, each of which is optionally substituted,
and [0795] Y.sup.C of ULM-g through ULM-i is N or C--R.sup.YC,
where R.sup.YC is H, OH, CN, NO.sub.2, halo (preferably Cl or F),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups (e.g. CF.sub.3), optionally substituted O(C.sub.1-C.sub.6
alkyl) (preferably substituted with one or two hydroxyl groups or
up to three halo groups) or an optionally substituted acetylenic
group --C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6
alkyl group (preferably C.sub.1-C.sub.3 alkyl), each of which
groups may be optionally connected to a PTM group (including a ULM'
group) via a linker group.
[0796] Preferred heterocycle groups for R.sup.2' of ULM-g through
ULM-i include tetrahydrofuran, tetrahydrothiene,
tetrahydroquinoline, piperidine, piperazine, pyrrollidine,
morpholine, oxane or thiane, each of which groups may be optionally
substituted, or a group according to the chemical structure:
##STR00252##
[0797] preferably, a
##STR00253##
group, wherein: [0798] R.sup.PRO of ULM-g through ULM-i is H,
optionally substituted C.sub.1-C.sub.6 alkyl or an optionally
substituted aryl, heteroaryl or heterocyclic group; [0799]
R.sup.PRO1 and R.sup.PRO2 of ULM-g through ULM-i are each
independently H, an optionally substituted C.sub.1-C.sub.3 alkyl
group or together form a keto group and [0800] each n of ULM-g
through ULM-i is independently 0, 1, 2, 3, 4, 5, or 6 (often 0 or
1), each of which groups may be optionally connected to a PTM group
(including a ULM' group) via a linker group.
[0801] Preferred R.sup.2' substituents of ULM-g through ULM-i also
include specifically (and without limitation to the specific
compound disclosed) the R.sup.2' substituents which are found in
the identified compounds disclosed herein (which includes the
specific compounds which are disclosed in the present
specification, and the figures which are attached hereto). Each of
these R.sup.2' substituents may be used in conjunction with any
number of R.sup.3' substituents which are also disclosed
herein.
[0802] R.sup.3' of ULM-g through ULM-i is preferably an optionally
substituted -T-Aryl, an optionally substituted-T-Heteroaryl, an
optionally substituted -T-Heterocycle, an optionally
substituted-NR.sup.1-T-Aryl, an optionally substituted
--NR.sup.1-T-Heteroaryl or an optionally
substituted-NR.sup.1-T-Heterocycle, where R.sup.1 is H or a
C.sub.1-C.sub.3 alkyl group, preferably H or CH.sub.3, T is an
optionally substituted --(CH.sub.2).sub.n-- group, wherein each one
of the methylene groups may be optionally substituted with one or
two substituents, preferably selected from halogen, a
C.sub.1-C.sub.3 alkyl group or the sidechain of an amino acid as
otherwise described herein, preferably methyl, which may be
optionally substituted; and n is 0 to 6, often 0, 1, 2, or 3
preferably 0 or 1. Alternatively, T may also be a
--(CH.sub.2O).sub.n-- group, a --(OCH.sub.2).sub.n-- group, a
--(CH.sub.2CH.sub.2O).sub.n-- group, a
--(OCH.sub.2CH.sub.2).sub.n-- group, each of which groups is
optionally substituted.
[0803] Preferred aryl groups for R.sup.3' of ULM-g through ULM-i
include optionally substituted phenyl or naphthyl groups,
preferably phenyl groups, wherein the phenyl or naphthyl group is
optionally connected to a PTM group (including a ULM' group) via a
linker group and/or optionally substituted with a halogen
(preferably F or Cl), an amine, monoalkyl- or dialkyl amine
(preferably, dimethylamine), an amido group (preferably a
--(CH.sub.2).sub.m--NR.sub.1C(O)R.sub.2 group where m, R.sub.1 and
R.sub.2 are the same as above), a halo (often F or Cl), OH,
CH.sub.3, CF.sub.3, OMe, OCF.sub.3, NO.sub.2, CN or a
S(O).sub.2R.sub.S group (R.sub.S is a C.sub.1-C.sub.6 alkyl group,
an optionally substituted aryl, heteroaryl or heterocycle group or
a --(CH.sub.2).sub.mNR.sub.1R.sub.2 group), each of which may be
substituted in ortho-, meta- and/or para-positions of the phenyl
ring, preferably para-), or an Aryl (preferably phenyl), Heteroaryl
or Heterocycle. Preferably said substituent phenyl group is an
optionally substituted phenyl group (i.e., the substituent phenyl
group itself is preferably substituted with at least one of F, Cl,
OH, SH, COOH, CH.sub.3, CF.sub.3, OMe, OCF.sub.3, NO.sub.2, CN or a
linker group to which is attached a PTM group (including a ULM'
group), wherein the substitution occurs in ortho-, meta- and/or
para-positions of the phenyl ring, preferably para-), a naphthyl
group, which may be optionally substituted including as described
above, an optionally substituted heteroaryl (preferably an
optionally substituted isoxazole including a methylsubstituted
isoxazole, an optionally substituted oxazole including a
methylsubstituted oxazole, an optionally substituted thiazole
including a methyl substituted thiazole, an optionally substituted
pyrrole including a methylsubstituted pyrrole, an optionally
substituted imidazole including a methylimidazole, a
benzylimidazole or methoxybenzylimidazole, an oximidazole or
methyloximidazole, an optionally substituted diazole group,
including a methyldiazole group, an optionally substituted triazole
group, including a methylsubstituted triazole group, a pyridine
group, including a halo-(preferably, F) or methyl substituted
pyridine group or an oxapyridine group (where the pyridine group is
linked to the phenyl group by an oxygen) or an optionally
substituted heterocycle (tetrahydrofuran, tetrahydrothiophene,
pyrrolidine, piperidine, morpholine, piperazine,
tetrahydroquinoline, oxane or thiane. Each of the aryl, heteroaryl
or heterocyclic groups may be optionally connected to a PTM group
(including a ULM' group) via a linker group.
[0804] Preferred Heteroaryl groups for R.sup.3' of ULM-g through
ULM-i include an optionally substituted quinoline (which may be
attached to the pharmacophore or substituted on any carbon atom
within the quinoline ring), an optionally substituted indole
(including dihydroindole), an optionally substituted indolizine, an
optionally substituted azaindolizine (2, 3 or 4-azaindolizine) an
optionally substituted benzimidazole, benzodiazole, benzoxofuran,
an optionally substituted imidazole, an optionally substituted
isoxazole, an optionally substituted oxazole (preferably methyl
substituted), an optionally substituted diazole, an optionally
substituted triazole, a tetrazole, an optionally substituted
benzofuran, an optionally substituted thiophene, an optionally
substituted thiazole (preferably methyl and/or thiol substituted),
an optionally substituted isothiazole, an optionally substituted
triazole (preferably a 1,2,3-triazole substituted with a methyl
group, a triisopropylsilyl group, an optionally substituted
--(CH.sub.2).sub.m--O--C.sub.1-C.sub.6 alkyl group or an optionally
substituted --(CH.sub.2).sub.m--C(O)--O--C.sub.1-C.sub.6 alkyl
group), an optionally substituted pyridine (2-, 3, or 4-pyridine)
or a group according to the chemical structure:
##STR00254##
wherein: [0805] S.sup.c of ULM-g through ULM-i is CHR.sup.SS,
NR.sup.URE, or O; [0806] R.sup.HET of ULM-g through ULM-i is H, CN,
NO.sub.2, halo (preferably Cl or F), optionally substituted
C.sub.1-C.sub.6 alkyl (preferably substituted with one or two
hydroxyl groups or up to three halo groups (e.g. CF.sub.3),
optionally substituted O(C.sub.1-C.sub.6 alkyl) (preferably
substituted with one or two hydroxyl groups or up to three halo
groups) or an optionally substituted acetylenic group
--C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6 alkyl
group (preferably C.sub.1-C.sub.3 alkyl); [0807] R.sup.SS of ULM-g
through ULM-i is H, CN, NO.sub.2, halo (preferably F or Cl),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups), optionally substituted O--(C.sub.1-C.sub.6 alkyl)
(preferably substituted with one or two hydroxyl groups or up to
three halo groups) or an optionally substituted
--C(O)(C.sub.1-C.sub.6alkyl) (preferably substituted with one or
two hydroxyl groups or up to three halo groups); [0808] R.sup.URE
of ULM-g through ULM-i is H, a C.sub.1-C.sub.6 alkyl (preferably H
or C.sub.1-C.sub.3 alkyl) or a --C(O)(C.sub.1-C.sub.6 alkyl), each
of which groups is optionally substituted with one or two hydroxyl
groups or up to three halogen, preferably fluorine groups, or an
optionally substituted heterocycle, for example piperidine,
morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene,
piperidine, piperazine, each of which is optionally substituted,
and [0809] Y.sup.C of ULM-g through ULM-i is N or C--R.sup.YC,
where R.sup.YC is H, OH, CN, NO.sub.2, halo (preferably Cl or F),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups (e.g. CF.sub.3), optionally substituted O(C.sub.1-C.sub.6
alkyl) (preferably substituted with one or two hydroxyl groups or
up to three halo groups) or an optionally substituted acetylenic
group --C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6
alkyl group (preferably C.sub.1-C.sub.3 alkyl). Each of said
heteroaryl groups may be optionally connected to a PTM group
(including a ULM' group) via a linker group.
[0810] Preferred heterocycle groups for R.sup.3' of ULM-g through
ULM-i include tetrahydroquinoline, piperidine, piperazine,
pyrrollidine, morpholine, tetrahydrofuran, tetrahydrothiophene,
oxane and thiane, each of which groups may be optionally
substituted or a group according to the chemical structure:
##STR00255##
preferably, a
##STR00256##
group, wherein: [0811] R.sup.PRO of ULM-g through ULM-i is H,
optionally substituted C.sub.1-C.sub.6 alkyl or an optionally
substituted aryl (phenyl or napthyl), heteroaryl or heterocyclic
group selected from the group consisting of oxazole, isoxazole,
thiazole, isothiazole, imidazole, diazole, oximidazole, pyrrole,
pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene,
dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine,
morpholine, quinoline, (each preferably substituted with a
C.sub.1-C.sub.3 alkyl group, preferably methyl or a halo group,
preferably F or Cl), benzofuran, indole, indolizine, azaindolizine;
[0812] R.sup.PRO1 and R.sup.PRO2 of ULM-g through ULM-i are each
independently H, an optionally substituted C.sub.1-C.sub.3 alkyl
group or together form a keto group, and [0813] each n of ULM-g
through ULM-i is 0, 1, 2, 3, 4, 5, or 6 (preferably 0 or 1),
wherein each of said Heterocycle groups may be optionally connected
to a PTM group (including a ULM' group) via a linker group.
[0814] Preferred R.sup.3' substituents of ULM-g through ULM-i also
include specifically (and without limitation to the specific
compound disclosed) the R.sup.3' substituents which are found in
the identified compounds disclosed herein (which includes the
specific compounds which are disclosed in the present
specification, and the figures which are attached hereto). Each of
these R.sup.3' substituents may be used in conjunction with any
number of R.sup.2' substituents, which are also disclosed
herein.
[0815] In certain alternative preferred embodiments, R.sup.2' of
ULM-g through ULM-i is an optionally substituted
--NR.sub.1--X.sup.R2'-alkyl group, --NR1-X.sup.R2'-Aryl group; an
optionally substituted --NR.sub.1--X.sup.R2'-HET, an optionally
substituted --NR1-X.sup.R2'-Aryl-HET or an optionally substituted
--NR.sub.1--X.sup.R2'-HET-Aryl,
wherein: [0816] R.sub.1 of ULM-g through ULM-i is H or a
C.sub.1-C.sub.3 alkyl group (preferably H); [0817] X.sup.R2' of
ULM-g through ULM-i is an optionally substituted
--CH.sub.2).sub.n--,
--CH.sub.2).sub.n--CH(X.sub.v).dbd.CH(X.sub.v)-- (cis or trans),
--(CH.sub.2).sub.n--CH.ident.CH--, --(CH.sub.2CH.sub.2O).sub.n-- or
a C.sub.3-C.sub.6 cycloalkyl group; and [0818] X.sub.v of ULM-g
through ULM-i is H, a halo or a C.sub.1-C.sub.3 alkyl group which
is optionally substituted with one or two hydroxyl groups or up to
three halogen groups; [0819] Alkyl of ULM-g through ULM-i is an
optionally substituted C1-C.sub.10 alkyl (preferably a
C.sub.1-C.sub.6 alkyl) group (in certain preferred embodiments, the
alkyl group is end-capped with a halo group, often a Cl or Br);
[0820] Aryl of ULM-g through ULM-i is an optionally substituted
phenyl or naphthyl group (preferably, a phenyl group); and [0821]
HET of ULM-g through ULM-i is an optionally substituted oxazole,
isoxazole, thiazole, isothiazole, imidazole, diazole, oximidazole,
pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene,
dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine,
morpholine, benzofuran, indole, indolizine, azaindolizine,
quinoline (when substituted, each preferably substituted with a
C.sub.1-C.sub.3 alkyl group, preferably methyl or a halo group,
preferably F or Cl) or a group according to the chemical
structure:
[0821] ##STR00257## [0822] S.sup.c of ULM-g through ULM-i is
CHR.sup.SS, NR.sup.URE, or O; [0823] R.sup.HET of ULM-g through
ULM-i is H, CN, NO.sub.2, halo (preferably Cl or F), optionally
substituted C.sub.1-C.sub.6 alkyl (preferably substituted with one
or two hydroxyl groups or up to three halo groups (e.g. CF.sub.3),
optionally substituted O(C.sub.1-C.sub.6 alkyl) (preferably
substituted with one or two hydroxyl groups or up to three halo
groups) or an optionally substituted acetylenic group
--C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6 alkyl
group (preferably C.sub.1-C.sub.3 alkyl); [0824] R.sup.SS of ULM-g
through ULM-i is H, CN, NO.sub.2, halo (preferably F or Cl),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups), optionally substituted O--(C.sub.1-C.sub.6 alkyl)
(preferably substituted with one or two hydroxyl groups or up to
three halo groups) or an optionally substituted
--C(O)(C.sub.1-C.sub.6alkyl) (preferably substituted with one or
two hydroxyl groups or up to three halo groups); [0825] R.sup.URE
of ULM-g through ULM-i is H, a C.sub.1-C.sub.6 alkyl (preferably H
or C.sub.1-C.sub.3 alkyl) or a --C(O)(C.sub.1-C.sub.6 alkyl), each
of which groups is optionally substituted with one or two hydroxyl
groups or up to three halogen, preferably fluorine groups, or an
optionally substituted heterocycle, for example piperidine,
morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene,
piperidine, piperazine, each of which is optionally substituted;
[0826] Y.sup.C of ULM-g through ULM-i is N or C--R.sup.YC, where
R.sup.YC is H, OH, CN, NO.sub.2, halo (preferably Cl or F),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups (e.g. CF.sub.3), optionally substituted O(C.sub.1-C.sub.6
alkyl) (preferably substituted with one or two hydroxyl groups or
up to three halo groups) or an optionally substituted acetylenic
group --C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6
alkyl group (preferably C.sub.1-C.sub.3 alkyl); [0827] R.sup.PRO of
ULM-g through ULM-i is H, optionally substituted C.sub.1-C.sub.6
alkyl or an optionally substituted aryl (phenyl or napthyl),
heteroaryl or heterocyclic group selected from the group consisting
of oxazole, isoxazole, thiazole, isothiazole, imidazole, diazole,
oximidazole, pyrrole, pyrollidine, furan, dihydrofuran,
tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine,
piperidine, piperazine, morpholine, quinoline, (each preferably
substituted with a C.sub.1-C.sub.3 alkyl group, preferably methyl
or a halo group, preferably F or Cl), benzofuran, indole,
indolizine, azaindolizine; [0828] R.sup.PRO1 and R.sup.PRO2 of
ULM-g through ULM-i are each independently H, an optionally
substituted C.sub.1-C.sub.3 alkyl group or together form a keto
group, and [0829] each n of ULM-g through ULM-i is independently 0,
1, 2, 3, 4, 5, or 6 (preferably 0 or 1).
[0830] Each of said groups may be optionally connected to a PTM
group (including a ULM' group) via a linker group.
[0831] In certain alternative preferred embodiments of the present
disclosure, R.sup.3' of ULM-g through ULM-i is an optionally
substituted
--(CH.sub.2).sub.n--(V).sub.n'--(CH2).sub.n-(V).sub.n'--R.sup.S3'
group, an optionally
substituted-(CH2).sub.n-N(R.sub.1')(C.dbd.O).sub.m'--(V).sub.n'--R.sup.S3-
' group, an optionally substituted --X.sup.R3'-alkyl group, an
optionally substituted --X.sup.R3'-Aryl group; an optionally
substituted --X.sup.R3'-HET group, an optionally substituted
--X.sup.R3'-Aryl-HET group or an optionally substituted
--X.sup.R3'-HET-Aryl group,
wherein: [0832] R.sup.S3' is an optionally substituted alkyl group
(C.sub.1-C.sub.10, preferably C.sub.1-C.sub.6 alkyl), an optionally
substituted Aryl group or a HET group; [0833] R.sub.1' is H or a
C.sub.1-C.sub.3 alkyl group (preferably H); [0834] V is O, S or
NR.sub.1'; [0835] X.sup.R3' is --(CH.sub.2).sub.n--,
--(CH.sub.2CH.sub.2O).sub.n--,
--CH.sub.2).sub.n--CH(X.sub.v).dbd.CH(X.sub.v)-- (cis or trans),
--CH.sub.2).sub.n--CH.ident.CH--, or a C.sub.3-C.sub.6 cycloalkyl
group, all optionally substituted; [0836] X.sub.v is H, a halo or a
C.sub.1-C.sub.3 alkyl group which is optionally substituted with
one or two hydroxyl groups or up to three halogen groups; [0837]
Alkyl is an optionally substituted C.sub.1-C.sub.10 alkyl
(preferably a C.sub.1-C.sub.6 alkyl) group (in certain preferred
embodiments, the alkyl group is end-capped with a halo group, often
a Cl or Br); [0838] Aryl is an optionally substituted phenyl or
napthyl group (preferably, a phenyl group); and [0839] HET is an
optionally substituted oxazole, isoxazole, thiazole, isothiazole,
imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan,
dihydrofuran, tetrahydrofuran, thiene, dihydrothiene,
tetrahydrothiene, pyridine, piperidine, piperazine, morpholine,
benzofuran, indole, indolizine, azaindolizine, quinoline (when
substituted, each preferably substituted with a C.sub.1-C.sub.3
alkyl group, preferably methyl or a halo group, preferably F or
Cl), or a group according to the chemical structure:
[0839] ##STR00258## [0840] S.sup.c of ULM-g through ULM-i is
CHR.sup.SS, NR.sup.URE, or O; [0841] R.sup.HET of ULM-g through
ULM-i is H, CN, NO.sub.2, halo (preferably Cl or F), optionally
substituted C.sub.1-C.sub.6 alkyl (preferably substituted with one
or two hydroxyl groups or up to three halo groups (e.g. CF.sub.3),
optionally substituted O(C.sub.1-C.sub.6 alkyl) (preferably
substituted with one or two hydroxyl groups or up to three halo
groups) or an optionally substituted acetylenic group
--C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6 alkyl
group (preferably C.sub.1-C.sub.3 alkyl); [0842] R.sup.SS of ULM-g
through ULM-i is H, CN, NO.sub.2, halo (preferably F or Cl),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups), optionally substituted O--(C.sub.1-C.sub.6 alkyl)
(preferably substituted with one or two hydroxyl groups or up to
three halo groups) or an optionally substituted
--C(O)(C.sub.1-C.sub.6alkyl) (preferably substituted with one or
two hydroxyl groups or up to three halo groups); [0843] R.sup.URE
of ULM-g through ULM-i is H, a C.sub.1-C.sub.6 alkyl (preferably H
or C.sub.1-C.sub.3 alkyl) or a --C(O)(C.sub.0-C.sub.6 alkyl), each
of which groups is optionally substituted with one or two hydroxyl
groups or up to three halogen, preferably fluorine groups, or an
optionally substituted heterocycle, for example piperidine,
morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene,
piperidine, piperazine, each of which is optionally substituted;
[0844] Y.sup.C of ULM-g through ULM-i is N or C--R.sup.YC, where
R.sup.YC is H, OH, CN, NO.sub.2, halo (preferably Cl or F),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups (e.g. CF.sub.3), optionally substituted O(C.sub.1-C.sub.6
alkyl) (preferably substituted with one or two hydroxyl groups or
up to three halo groups) or an optionally substituted acetylenic
group --C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6
alkyl group (preferably C.sub.1-C.sub.3 alkyl); [0845] R.sup.PRO of
ULM-g through ULM-i is H, optionally substituted C.sub.1-C.sub.6
alkyl or an optionally substituted aryl (phenyl or napthyl),
heteroaryl or heterocyclic group selected from the group consisting
of oxazole, isoxazole, thiazole, isothiazole, imidazole, diazole,
oximidazole, pyrrole, pyrollidine, furan, dihydrofuran,
tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine,
piperidine, piperazine, morpholine, quinoline, (each preferably
substituted with a C.sub.1-C.sub.3 alkyl group, preferably methyl
or a halo group, preferably F or Cl), benzofuran, indole,
indolizine, azaindolizine; [0846] R.sup.PRO1 and R.sup.PRO2 of
ULM-g through ULM-i are each independently H, an optionally
substituted C.sub.1-C.sub.3 alkyl group or together form a keto
group; [0847] each n of ULM-g through ULM-i is independently 0, 1,
2, 3, 4, 5, or 6 (preferably 0 or 1); [0848] each m' of ULM-g
through ULM-i is 0 or 1; and [0849] each n' of ULM-g through ULM-i
is 0 or 1; [0850] wherein each of said compounds, preferably on the
alkyl, Aryl or Het groups, is optionally connected to a PTM group
(including a ULM' group) via a linker.
[0851] In alternative embodiments, R.sup.3' of ULM-g through ULM-i
is --(CH.sub.2).sub.n-Aryl, --(CH.sub.2CH.sub.2O).sub.n-Aryl,
--(CH.sub.2).sub.n-HET or --(CH.sub.2CH.sub.2O).sub.n-HET,
wherein: [0852] said Aryl of ULM-g through ULM-i is phenyl which is
optionally substituted with one or two substitutents, wherein said
substituent(s) is preferably selected from --(CH.sub.2).sub.nOH,
C.sub.1-C.sub.6 alkyl which itself is further optionally
substituted with CN, halo (up to three halo groups), OH,
--(CH.sub.2).sub.nO(C.sub.1-C.sub.6)alkyl, amine, mono- or
di-(C.sub.1-C.sub.6 alkyl) amine wherein the alkyl group on the
amine is optionally substituted with 1 or 2 hydroxyl groups or up
to three halo (preferably F, Cl) groups, or [0853] said Aryl group
of ULM-g through ULM-i is substituted with --(CH.sub.2).sub.nOH,
--(CH.sub.2).sub.n--O--(C.sub.1-C.sub.6)alkyl,
--(CH.sub.2).sub.n--O--(CH.sub.2).sub.n--(C.sub.1-C.sub.6)alkyl,
--(CH.sub.2).sub.n--C(O)(C.sub.0-C.sub.6) alkyl,
--(CH.sub.2).sub.n--C(O)O(C.sub.0-C.sub.6)alkyl,
--(CH.sub.2).sub.n--OC(O)(C.sub.0-C.sub.6)alkyl, amine, mono- or
di-(C.sub.1-C.sub.6 alkyl) amine wherein the alkyl group on the
amine is optionally substituted with 1 or 2 hydroxyl groups or up
to three halo (preferably F, Cl) groups, CN, NO.sub.2, an
optionally substituted
--(CH.sub.2).sub.n--(V).sub.m'--CH.sub.2).sub.n--(V).sub.m'--(C.sub.1-C.s-
ub.6)alkyl group, a
--(V).sub.m'--(CH.sub.2CH.sub.2O).sub.n--R.sup.PEG group where V is
O, S or NR.sub.1', R.sub.1' is H or a C.sub.1-C.sub.3 alkyl group
(preferably H) and R.sup.PEG is H or a C.sub.1-C.sub.6 alkyl group
which is optionally substituted (including being optionally
substituted with a carboxyl group), or [0854] said Aryl group of
ULM-g through ULM-i is optionally substituted with a heterocycle,
including a heteroaryl, selected from the group consisting of
oxazole, isoxazole, thiazole, isothiazole, imidazole, diazole,
oximidazole, pyrrole, pyrollidine, furan, dihydrofuran,
tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine,
piperidine, piperazine, morpholine, quinoline, benzofuran, indole,
indolizine, azaindolizine, (when substituted each preferably
substituted with a C.sub.1-C.sub.3 alkyl group, preferably methyl
or a halo group, preferably F or Cl), or a group according to the
chemical structure:
[0854] ##STR00259## [0855] S.sup.c of ULM-g through ULM-i is
CHR.sup.SS, NR.sup.URE, or O; [0856] R.sup.HET of ULM-g through
ULM-i is H, CN, NO.sub.2, halo (preferably Cl or F), optionally
substituted C.sub.1-C.sub.6 alkyl (preferably substituted with one
or two hydroxyl groups or up to three halo groups (e.g. CF.sub.3),
optionally substituted O(C.sub.1-C.sub.6 alkyl) (preferably
substituted with one or two hydroxyl groups or up to three halo
groups) or an optionally substituted acetylenic group
--C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6 alkyl
group (preferably C.sub.1-C.sub.3 alkyl); [0857] R.sup.SS of ULM-g
through ULM-i is H, CN, NO.sub.2, halo (preferably F or Cl),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups), optionally substituted O--(C.sub.1-C.sub.6 alkyl)
(preferably substituted with one or two hydroxyl groups or up to
three halo groups) or an optionally substituted
--C(O)(C.sub.1-C.sub.6alkyl) (preferably substituted with one or
two hydroxyl groups or up to three halo groups); [0858] R.sup.URE
of ULM-g through ULM-i is H, a C.sub.1-C.sub.6 alkyl (preferably H
or C.sub.1-C.sub.3 alkyl) or a --C(O)(C.sub.0-C.sub.6 alkyl), each
of which groups is optionally substituted with one or two hydroxyl
groups or up to three halogen, preferably fluorine groups, or an
optionally substituted heterocycle, for example piperidine,
morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene,
piperidine, piperazine, each of which is optionally substituted;
[0859] Y.sup.C of ULM-g through ULM-i is N or C--R.sup.YC, where
R.sup.YC is H, OH, CN, NO.sub.2, halo (preferably Cl or F),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups (e.g. CF.sub.3), optionally substituted O(C.sub.1-C.sub.6
alkyl) (preferably substituted with one or two hydroxyl groups or
up to three halo groups) or an optionally substituted acetylenic
group --C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6
alkyl group (preferably C.sub.1-C.sub.3 alkyl); [0860] R.sup.PRO of
ULM-g through ULM-i is H, optionally substituted C.sub.1-C.sub.6
alkyl or an optionally substituted aryl (phenyl or napthyl),
heteroaryl or heterocyclic group selected from the group consisting
of oxazole, isoxazole, thiazole, isothiazole, imidazole, diazole,
oximidazole, pyrrole, pyrollidine, furan, dihydrofuran,
tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine,
piperidine, piperazine, morpholine, quinoline, (each preferably
substituted with a C.sub.1-C.sub.3 alkyl group, preferably methyl
or a halo group, preferably F or Cl), benzofuran, indole,
indolizine, azaindolizine; [0861] R.sup.PRO1 and R.sup.PRO2 of
ULM-g through ULM-i are each independently H, an optionally
substituted C.sub.1-C.sub.3 alkyl group or together form a keto
group; [0862] HET of ULM-g through ULM-i is preferably oxazole,
isoxazole, thiazole, isothiazole, imidazole, diazole, oximidazole,
pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene,
dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine,
morpholine, quinoline, (each preferably substituted with a
C.sub.1-C.sub.3 alkyl group, preferably methyl or a halo group,
preferably F or Cl), benzofuran, indole, indolizine, azaindolizine,
or a group according to the chemical structure:
[0862] ##STR00260## [0863] S.sup.c of ULM-g through ULM-i is
CHR.sup.SS, NR.sup.URE, or O; [0864] R.sup.HET of ULM-g through
ULM-i is H, CN, NO.sub.2, halo (preferably Cl or F), optionally
substituted C.sub.1-C.sub.6 alkyl (preferably substituted with one
or two hydroxyl groups or up to three halo groups (e.g. CF.sub.3),
optionally substituted O(C.sub.1-C.sub.6 alkyl) (preferably
substituted with one or two hydroxyl groups or up to three halo
groups) or an optionally substituted acetylenic group
--C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6 alkyl
group (preferably C.sub.1-C.sub.3 alkyl); [0865] R.sup.SS of ULM-g
through ULM-i is H, CN, NO.sub.2, halo (preferably F or Cl),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups), optionally substituted O--(C.sub.1-C.sub.6 alkyl)
(preferably substituted with one or two hydroxyl groups or up to
three halo groups) or an optionally substituted
--C(O)(C.sub.1-C.sub.6alkyl) (preferably substituted with one or
two hydroxyl groups or up to three halo groups); [0866] R.sup.URE
of ULM-g through ULM-i is H, a C.sub.1-C.sub.6 alkyl (preferably H
or C.sub.1-C.sub.3 alkyl) or a --C(O)(C.sub.0-C.sub.6 alkyl), each
of which groups is optionally substituted with one or two hydroxyl
groups or up to three halogen, preferably fluorine groups, or an
optionally substituted heterocycle, for example piperidine,
morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene,
piperidine, piperazine, each of which is optionally substituted;
[0867] Y.sup.C of ULM-g through ULM-i is N or C--R.sup.YC, where
R.sup.YC is H, OH, CN, NO.sub.2, halo (preferably Cl or F),
optionally substituted C.sub.1-C.sub.6 alkyl (preferably
substituted with one or two hydroxyl groups or up to three halo
groups (e.g. CF.sub.3), optionally substituted O(C.sub.1-C.sub.6
alkyl) (preferably substituted with one or two hydroxyl groups or
up to three halo groups) or an optionally substituted acetylenic
group --C.ident.C--R.sub.a where R.sub.a is H or a C.sub.1-C.sub.6
alkyl group (preferably C.sub.1-C.sub.3 alkyl); [0868] R.sup.PRO of
ULM-g through ULM-i is H, optionally substituted C.sub.1-C.sub.6
alkyl or an optionally substituted aryl, heteroaryl or heterocyclic
group; [0869] R.sup.PRO1 and R.sup.PRO2 of ULM-g through ULM-i are
each independently H, an optionally substituted C.sub.1-C.sub.3
alkyl group or together form a keto group; [0870] each m' of ULM-g
through ULM-i is independently 0 or 1; and [0871] each n of ULM-g
through ULM-i is independently 0, 1, 2, 3, 4, 5, or 6 (preferably 0
or 1), [0872] wherein each of said compounds, preferably on said
Aryl or HET groups, is optionally connected to a PTM group
(including a ULM' group) via a linker group.
[0873] In still additional embodiments, preferred compounds include
those according to the chemical structure:
##STR00261##
wherein: [0874] R.sup.1' of ULM-i is OH or a group which is
metabolized in a patient or subject to OH; [0875] R.sup.2' of ULM-i
is a --NH--CH.sub.2-Aryl-HET (preferably, a phenyl linked directly
to a methyl substituted thiazole); [0876] R.sup.3' of ULM-i is a
--CHR.sup.CR3'--NH--C(O)--R.sup.3P1 group or a
--CHR.sup.CR3'--R.sup.3P2 group; [0877] R.sup.CR3' of ULM-i is a
C.sub.1-C.sub.4 alkyl group, preferably methyl, isopropyl or
tert-butyl; [0878] R.sup.3P1 of ULM-i is C.sub.1-C.sub.3 alkyl
(preferably methyl), an optionally substituted oxetane group
(preferably methyl substituted, a --(CH.sub.2).sub.nOCH.sub.3 group
where n is 1 or 2 (preferably 2), or a
##STR00262##
[0878] group (the ethyl ether group is preferably meta-substituted
on the phenyl moiety), a morpholino group (linked to the carbonyl
at the 2- or 3-position; [0879] R.sup.3P2 of ULM-i is a
##STR00263##
[0879] group; [0880] Aryl of ULM-i is phenyl; [0881] HET of ULM-i
is an optionally substituted thiazole or isothiazole; and [0882]
R.sup.HET of ULM-i is H or a halo group (preferably H); [0883] or a
pharmaceutically acceptable salt, stereoisomer, solvate or
polymorph thereof, wherein each of said compounds is optionally
connected to a PTM group (including a ULM' group) via a linker
group.
[0884] In certain aspects, bifunctional compounds comprising a
ubiquitin E3 ligase binding moiety (ULM), wherein ULM is a group
according to the chemical structure:
##STR00264##
wherein: each R.sub.5 and R.sub.6 of ULM-j is independently OH, SH,
or optionally substituted alkyl or R.sub.5, R.sub.6, and the carbon
atom to which they are attached form a carbonyl; R.sub.7 of ULM-j
is H or optionally substituted alkyl; E of ULM-j is a bond,
C.dbd.O, or C.dbd.S; G of ULM-j is a bond, optionally substituted
alkyl, --COOH or C=J;
J of ULM-j is O or N--R8;
[0885] R.sub.8 of ULM-j is H, CN, optionally substituted alkyl or
optionally substituted alkoxy; M of ULM-j is optionally substituted
aryl, optionally substituted heteroaryl, optionally substituted
heterocyclic or
##STR00265##
each R.sub.9 and R.sub.10 of ULM-j is independently H; optionally
substituted alkyl, optionally substituted cycloalkyl, optionally
substituted hydroxyalkyl, optionally substituted thioalkyl, a
disulphide linked ULM, optionally substituted heteroaryl, or
haloalkyl; or R.sub.9, R.sub.10, and the carbon atom to which they
are attached form an optionally substituted cycloalkyl; R.sub.11 of
ULM-j is optionally substituted heterocyclic, optionally
substituted alkoxy, optionally substituted heteroaryl, optionally
substituted aryl, or
##STR00266##
R.sub.12 of ULM-j is H or optionally substituted alkyl; R.sub.13 of
ULM-j is H, optionally substituted alkyl, optionally substituted
alkylcarbonyl, optionally substituted (cycloalkyl)alkylcarbonyl,
optionally substituted aralkylcarbonyl, optionally substituted
arylcarbonyl, optionally substituted (heterocyclyl)carbonyl, or
optionally substituted aralkyl; optionally substituted
(oxoalkyl)carbamate, each R.sub.14 of ULM-j is independently H,
haloalkyl, optionally substituted cycloalkyl, optionally
substituted alkyl or optionally substituted heterocycloalkyl;
R.sub.15 of ULM-j is H, optionally substituted heteroaryl,
haloalkyl, optionally substituted aryl, optionally substituted
alkoxy, or optionally substituted heterocyclyl; each R.sub.16 of
ULM-j is independently halo, optionally substituted alkyl,
optionally substituted haloalkyl, CN, or optionally substituted
haloalkoxy; each R.sub.25 of ULM-j is independently H or optionally
substituted alkyl; or both R.sub.25 groups can be taken together to
form an oxo or optionally substituted cycloalkyl group;
R.sub.23 of ULM-j is H or OH;
[0886] Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 of ULM-j are
independently C or N; and o of ULM-j is 0, 1, 2, 3, or 4, or a
pharmaceutically acceptable salt, stereoisomer, solvate or
polymorph thereof.
[0887] In certain embodiments, wherein G of ULM-j is C=J, J is O,
R.sub.7 is H, each R.sub.14 is H, and o is 0.
[0888] In certain embodiments, wherein G of ULM-j is C=J, J is O,
R.sub.7 is H, each R.sub.14 is H, R.sub.15 is optionally
substituted heteroaryl, and o is O. In other instances, E is
C.dbd.O and M is
##STR00267##
[0889] In certain embodiments, wherein E of ULM-j is C.dbd.O,
R.sub.11 is optionally substituted heterocyclic or
##STR00268##
and M is
##STR00269##
[0891] In certain embodiments, wherein E of ULM-j is C.dbd.O, M
is
##STR00270##
and R.sub.11 is
##STR00271##
[0892] each R.sub.18 is independently halo, optionally substituted
alkoxy, cyano, optionally substituted alkyl, haloalkyl, or
haloalkoxy; and p is 0, 1, 2, 3, or 4.
[0893] In certain embodiments, ULM and where present, ULM', are
each independently a group according to the chemical structure:
##STR00272##
wherein: [0894] G of ULM-k is C=J, J is O; [0895] R.sub.7 of ULM-k
is H; [0896] each R.sub.14 of ULM-k is H; [0897] o of ULM-k is O;
[0898] R.sub.15 of ULM-k is
##STR00273##
[0898] and [0899] R.sub.17 of ULM-k is H, halo, optionally
substituted cycloalkyl, optionally substituted alkyl, optionally
substituted alkenyl, and haloalkyl.
[0900] In other instances, R.sub.17 of ULM-k is alkyl (e.g.,
methyl) or cycloalkyl (e.g., cyclopropyl).
[0901] In other embodiments, ULM and where present, ULM', are each
independently a group according to the chemical structure:
##STR00274##
wherein:
[0902] G of ULM-k is C=J, J is O;
[0903] R.sub.7 of ULM-k is H;
[0904] each R.sub.14 of ULM-k is H;
[0905] o of ULM-k is 0; and
[0906] R.sub.15 of ULM-k is selected from the group consisting
of:
##STR00275##
wherein R.sub.30 of ULM-k is H or an optionally substituted
alkyl.
[0907] In other embodiments, ULM and where present, ULM', are each
independently a group according to the chemical structure:
##STR00276##
wherein:
[0908] E of ULM-k is C.dbd.O;
[0909] M of ULM-k is
##STR00277##
and
[0910] R.sub.11 of ULM-k is selected from the group consisting
of:
##STR00278##
[0911] In still other embodiments, a compound of the chemical
structure,
##STR00279##
wherein E of ULM-k is C.dbd.O;
R.sub.11 of ULM-k is
##STR00280##
[0912] and
M of ULM-k is
##STR00281##
[0913] q of ULM-k is 1 or 2;
[0914] R.sub.20 of ULM-k is H, optionally substituted alkyl,
optionally substituted cycloalkyl, optionally substituted aryl,
or
##STR00282##
R.sub.21 of ULM-k is H or optionally substituted alkyl; and
R.sub.22 of ULM-k is H, optionally substituted alkyl, optionally
substituted alkoxy, or haloalkyl.
[0915] In any embodiment described herein, R.sub.11 of ULM-j or
ULM-k is selected from the group consisting of:
##STR00283## ##STR00284## ##STR00285## ##STR00286##
[0916] In certain embodiments, R.sub.11 of ULM-j or ULM-k is
selected from the group consisting of:
##STR00287## ##STR00288## ##STR00289##
[0917] In certain embodiments, ULM (or when present ULM') is a
group according to the chemical structure:
##STR00290##
wherein: [0918] X of ULM-l is O or S; [0919] Y of ULM-l is H,
methyl or ethyl; [0920] R.sub.17 of ULM-l is H, methyl, ethyl,
hydoxymethyl or cyclopropyl; [0921] M of ULM-l is optionally
substituted aryl, optionally substituted heteroaryl, or
[0921] ##STR00291## [0922] R.sub.9 of ULM-l is H; [0923] R.sub.10
of ULM-l is H, optionally substituted alkyl, optionally substituted
haloalkyl, optionally substituted heteroaryl, optionally
substituted aryl, optionally substituted hydroxyalkyl, optionally
substituted thioalkyl or cycloalkyl; [0924] R.sub.11 of ULM-l is
optionally substituted heteroaromatic, optionally substituted
heterocyclic, optionally substituted aryl or
[0924] ##STR00292## [0925] R.sub.12 of ULM-l is H or optionally
substituted alkyl; and [0926] R.sub.13 of ULM-l is H, optionally
substituted alkyl, optionally substituted alkylcarbonyl, optionally
substituted (cycloalkyl)alkylcarbonyl, optionally substituted
aralkylcarbonyl, optionally substituted arylcarbonyl, optionally
substituted (heterocyclyl)carbonyl, or optionally substituted
aralkyl; optionally substituted (oxoalkyl)carbamate.
[0927] In some embodiments, ULM and where present, ULM', are each
independently a group according to the chemical structure:
##STR00293##
wherein: [0928] Y of ULM-m is H, methyol or ethyl [0929] R.sub.9 of
ULM-m is H; [0930] R.sub.10 is isopropyl, tert-butyl, sec-butyl,
cyclopentyl, or cyclohexyl; [0931] R.sub.11 of ULM-m is optionally
substituted amide, optionally substituted isoindolinone, optionally
substituted isooxazole, optionally substituted heterocycles.
[0932] In other preferred embodiments of the disclosure, ULM and
where present, ULM', are each independently a group according to
the chemical structure:
##STR00294##
wherein: [0933] R.sub.17 of ULM-n is methyl, ethyl, or cyclopropyl;
and [0934] R.sub.9, R.sub.10, and R.sub.11 of ULM-n are as defined
above. In other instances, R.sub.9 is H; and [0935] R.sub.10 of
ULM-n is H, alkyl, or cycloalkyl (preferably, isopropyl,
tert-butyl, sec-butyl, cyclopentyl, or cyclohexyl).
[0936] In any of the aspects or embodiments described herein, the
ULM (or when present, ULM') as described herein may be a
pharmaceutically acceptable salt, enantiomer, diastereomer, solvate
or polymorph thereof. In addition, in any of the aspects or
embodiments described herein, the ULM (or when present, ULM') as
described herein may be coupled to a PTM directly via a bond or by
a chemical linker.
[0937] In certain aspects of the disclosure, the ULM moiety is
selected from the group consisting of:
##STR00295## ##STR00296## ##STR00297## ##STR00298## ##STR00299##
##STR00300## ##STR00301## ##STR00302## ##STR00303## ##STR00304##
##STR00305## ##STR00306## ##STR00307## ##STR00308## ##STR00309##
##STR00310## ##STR00311## ##STR00312## ##STR00313## ##STR00314##
##STR00315## ##STR00316## ##STR00317##
##STR00318## ##STR00319## ##STR00320## ##STR00321## ##STR00322##
##STR00323## ##STR00324## ##STR00325## ##STR00326## ##STR00327##
##STR00328## ##STR00329## ##STR00330##
wherein the VLM may be connected to a PTM via a linker, as
described herein, at any appropriate location, including, e.g., an
aryl, heteroaryl, phenyl, or phenyl of an indole group, optionally
via any appropriate functional group, such as an amine, ester,
ether, alkyl, or alkoxy.
Exemplary Linkers
[0938] In certain embodiments, the compounds as described herein
include one or more PTMs chemically linked or coupled to one or
more ULMs (e.g., at least one of CLM, VLM, MLM, ILM, or a
combination thereof) via a chemical linker (L). In certain
embodiments, the linker group L is a group comprising one or more
covalently connected structural units (e.g., -A.sup.L.sub.1 . . .
(A.sup.L).sub.q- or -(A.sup.L).sub.q-), wherein A.sub.1 is a group
coupled to PTM, and (A.sup.L).sub.q is a group coupled to ULM.
[0939] In certain embodiments, the linker group L is selected from
-(A.sup.L).sub.q:
[0940] (A.sup.L).sub.q is a group which is connected to at least
one of a ULM, a PTM moiety, or a combination thereof;
[0941] q of the linker is an integer greater than or equal to
1;
[0942] each A.sup.L.sub.q is independently selected from the group
consisting of, a bond, CR.sup.L1R.sup.L2, O, S, SO, SO.sub.2,
NR.sup.L3, SO.sub.2NR.sup.L3, SONR.sup.L3, CONR.sup.L3,
NR.sup.L3CONR.sup.L4, NR.sup.L3SO.sub.2NR.sup.L4, CO,
CR.sup.L1.dbd.CR.sup.L2, C.ident.C, SiR.sup.L1R.sup.L2,
P(O)R.sup.L1, P(O)OR.sup.L1, NR.sup.L3C(.dbd.NCN)NR.sup.L4,
NR.sup.L3C(.dbd.NCN), NR.sup.L3C(.dbd.CNO.sub.2)NR.sup.L4,
C.sub.3-11cycloalkyl optionally substituted with 0-6 R.sup.L1
and/or R.sup.L2 groups, C.sub.5-13 spirocycloalkyl optionally
substituted with 0-9 R.sup.L1 and/or R.sup.L2 groups,
C.sub.3-11heterocyclyl optionally substituted with 0-6 R.sup.L1
and/or R.sup.L2 groups, C.sub.5-13 spiroheterocycloalkyl optionally
substituted with 0-8 R.sup.L1 and/or R.sup.L2 groups, aryl
optionally substituted with 0-6 R.sup.L1 and/or R.sup.L2 groups,
heteroaryl optionally substituted with 0-6 R.sup.L1 and/or R.sup.L2
groups, where R.sup.L1 or R.sup.L2, each independently are
optionally linked to other groups to form cycloalkyl and/or
heterocyclyl moiety, optionally substituted with 0-4 R.sup.L5
groups; and
[0943] R.sup.L1, R.sup.L2, R.sup.L3, R.sup.L4 and R.sup.L5 are,
each independently, H, halo, C.sub.1-8alkyl, OC.sub.1-8alkyl,
SC.sub.1-8alkyl, NHC.sub.1-8alkyl, N(C.sub.1-8alkyl).sub.2,
C.sub.3-11cycloalkyl, aryl, heteroaryl, C.sub.3-11heterocyclyl,
OC.sub.1-8cycloalkyl, SC.sub.1-8cycloalkyl, NHC.sub.1-8cycloalkyl,
N(C.sub.1-8cycloalkyl).sub.2,
N(C.sub.1-8cycloalkyl)(C.sub.1-8alkyl), OH, NH.sub.2, SH,
SO.sub.2C.sub.1-8alkyl, P(O)(OC.sub.1-8alkyl)(C.sub.1-8alkyl),
P(O)(OC.sub.1-8alkyl).sub.2, CC--C.sub.1-8alkyl, CCH,
CH.dbd.CH(C.sub.1-8alkyl),
C(C.sub.1-8alkyl).dbd.CH(C.sub.1-8alkyl),
C(C.sub.1-8alkyl).dbd.C(C.sub.1-8alkyl).sub.2, Si(OH).sub.3,
Si(C.sub.1-8alkyl).sub.3, Si(OH)(C.sub.1-8alkyl).sub.2,
COC.sub.1-8alkyl, CO.sub.2H, halogen, CN, CF.sub.3, CHF.sub.2,
CH.sub.2F, NO.sub.2, SF.sub.5, SO.sub.2NHC.sub.1-8alkyl,
SO.sub.2N(C.sub.1-8alkyl).sub.2, SONHC.sub.1-8alkyl,
SON(C.sub.1-8alkyl).sub.2, CONHC.sub.1-8alkyl,
CON(C.sub.1-8alkyl).sub.2, N(C.sub.1-8alkyl)CONH(C.sub.1-8alkyl),
N(C.sub.1-8alkyl)CON(C.sub.1-8alkyl).sub.2, NHCONH(C.sub.1-8alkyl),
NHCON(C.sub.1-8alkyl).sub.2, NHCONH.sub.2,
N(C.sub.1-8alkyl)SO.sub.2NH(C.sub.1-8alkyl), N(C.sub.1-8alkyl)
SO.sub.2N(C.sub.1-8alkyl).sub.2, NH SO.sub.2NH(C.sub.1-8alkyl), NH
SO.sub.2N(C.sub.1-8alkyl).sub.2, NH SO.sub.2NH.sub.2.
[0944] In certain embodiments, q of the linker is an integer
greater than or equal to 0. In certain embodiments, q is an integer
greater than or equal to 1.
[0945] In certain embodiments, e.g., where q of the linker is
greater than 2, (A.sup.L).sub.q is a group which is connected to
ULM, and A.sup.L.sub.1 and (A.sup.L).sub.q are connected via
structural units of the linker (L).
[0946] In certain embodiments, e.g., where q of the linker is 2,
(A.sup.L).sub.q is a group which is connected to A.sup.L.sub.1 and
to a ULM.
[0947] In certain embodiments, e.g., where q of the linker is 1,
the structure of the linker group L is -A.sup.L.sub.1-, and
A.sup.L.sub.1 is a group which is connected to a ULM moiety and a
PTM moiety.
[0948] In certain embodiments, the linker (L) comprises a group
represented by a general structure selected from the group
consisting of:
[0949] --NR(CH.sub.2).sub.n-(lower alkyl)-,
--NR(CH.sub.2).sub.n-(lower alkoxyl)-, --NR(CH.sub.2).sub.n-(lower
alkoxyl)-OCH.sub.2--, --NR(CH.sub.2).sub.n-(lower alkoxyl)-(lower
alkyl)-OCH.sub.2--, --NR(CH.sub.2).sub.n-(cycloalkyl)-(lower
alkyl)-OCH.sub.2-, --NR(CH.sub.2).sub.n-(hetero cycloalkyl)-,
--NR(CH.sub.2CH.sub.2O).sub.n-(lower alkyl)-O--CH.sub.2--,
--NR(CH.sub.2CH.sub.2O).sub.n-(hetero cycloalkyl)-O--CH.sub.2--,
--NR(CH.sub.2CH.sub.2O).sub.n-Aryl-O--CH.sub.2--,
--NR(CH.sub.2CH.sub.2O).sub.n-(hetero aryl)-O--CH.sub.2--,
--NR(CH.sub.2CH.sub.2O).sub.n-(cyclo alkyl)-O-(hetero
aryl)-O--CH.sub.2--, --NR(CH.sub.2CH.sub.2O).sub.n-(cyclo
alkyl)-O-Aryl-O--CH.sub.2--, --NR(CH.sub.2CH.sub.2O).sub.n-(lower
alkyl)-NH-Aryl-O--CH.sub.2--, --NR(CH.sub.2CH.sub.2O).sub.n-(lower
alkyl)-O-Aryl-CH.sub.2,
--NR(CH.sub.2CH.sub.2O).sub.n-cycloalkyl-O-Aryl-,
--NR(CH.sub.2CH.sub.2O).sub.n-cycloalkyl-O-(hetero aryl)l-,
--NR(CH2CH2)n-(cycloalkyl)-O-(heterocycle)-CH.sub.2,
--NR(CH2CH2)n-(heterocycle)-(heterocycle)-CH.sub.2,
--N(R1R2)-(heterocycle)-CH2; where
[0950] n of the linker can be 0 to 10;
[0951] R of the linker can be H, lower alkyl;
[0952] R1 and R2 of the linker can form a ring with the connecting
N.
[0953] In certain embodiments, the linker (L) comprises a group
represented by a general structure selected from the group
consisting of:
[0954]
--N(R)--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2)-
.sub.q--O(CH2).sub.r-OCH2-,
[0955]
--O--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2).su-
b.q--O(CH2).sub.r-OCH2-,
[0956]
--O--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2).su-
b.q--O(CH2).sub.r-O--;
[0957]
--N(R)--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2)-
.sub.q--O(CH2).sub.r-O--;
[0958]
--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2).sub.q-
--O(CH2).sub.r-O--;
[0959]
--(CH2).sub.m-O(CH2).sub.n-O(CH2).sub.o--O(CH2).sub.p--O(CH2).sub.q-
--O(CH2).sub.r-OCH2-;
##STR00331## ##STR00332## ##STR00333##
wherein
[0960] m, n, o, p, q, and r of the linker are independently 0, 1,
2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,
20;
[0961] when the number is zero, there is no N--O or O--O bond
[0962] R of the linker is H, methyl and ethyl;
[0963] X of the linker is H and F
##STR00334##
[0964] where m of the linker can be 2, 3, 4, 5
##STR00335## ##STR00336## ##STR00337## ##STR00338## ##STR00339##
##STR00340##
##STR00341## ##STR00342## ##STR00343## ##STR00344## ##STR00345##
##STR00346## ##STR00347## ##STR00348## ##STR00349## ##STR00350##
##STR00351## ##STR00352##
[0965] In any aspect or embodiment described herein, the linker (L)
is selected from the group consisting of:
##STR00353## ##STR00354## ##STR00355## ##STR00356## ##STR00357##
##STR00358## ##STR00359##
wherein each m and n is independently selected from 0, 1, 2, 3, 4,
5, or 6.
[0966] In any aspect or embodiment described herein, the linker (L)
is selected from the group consisting of:
##STR00360## ##STR00361## ##STR00362## ##STR00363## ##STR00364##
##STR00365## ##STR00366## ##STR00367## ##STR00368## ##STR00369##
##STR00370## ##STR00371## ##STR00372## ##STR00373## ##STR00374##
##STR00375## ##STR00376## ##STR00377## ##STR00378## ##STR00379##
##STR00380##
##STR00381## ##STR00382## ##STR00383## ##STR00384## ##STR00385##
##STR00386## ##STR00387## ##STR00388## ##STR00389## ##STR00390##
##STR00391## ##STR00392## ##STR00393## ##STR00394## ##STR00395##
##STR00396## ##STR00397##
##STR00398## ##STR00399## ##STR00400## ##STR00401## ##STR00402##
##STR00403## ##STR00404## ##STR00405## ##STR00406## ##STR00407##
##STR00408## ##STR00409## ##STR00410## ##STR00411##
wherein each m, n, o, p, q, and r is independently 0, 1, 2, 3, 4,
5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
[0967] In any aspect or embodiment described herein, L is selected
from the group consisting of:
##STR00412## ##STR00413## ##STR00414## ##STR00415## ##STR00416##
##STR00417## ##STR00418## ##STR00419## ##STR00420##
##STR00421##
##STR00422## ##STR00423## ##STR00424## ##STR00425## ##STR00426##
##STR00427## ##STR00428## ##STR00429## ##STR00430## ##STR00431##
##STR00432## ##STR00433## ##STR00434## ##STR00435## ##STR00436##
##STR00437## ##STR00438## ##STR00439## ##STR00440## ##STR00441##
##STR00442##
[0968] In additional embodiments, the linker (L) comprises a
structure selected from, but not limited to the structure shown
below, where a dashed line indicates the attachment point to the
PTM or ULM moieties:
##STR00443##
wherein: [0969] W.sup.L1 and W.sup.L2 are each independently a 4-8
membered ring with 0-4 heteroatoms, optionally substituted with
R.sup.Q, each R.sup.Q is independently a H, halo, OH, CN, CF.sub.3,
C.sub.1-C.sub.6 alkyl (linear, branched, optionally substituted),
C.sub.1-C.sub.6 alkoxy (linear, branched, optionally substituted),
or 2 R.sup.Q groups taken together with the atom they are attached
to, form a 4-8 membered ring system containing 0-4 heteroatoms;
[0970] Y.sup.L1 is each independently a bond, C.sub.1-C.sub.6 alkyl
(linear, branched, optionally substituted) and optionally one or
more C atoms are replaced with O; or C.sub.1-C.sub.6 alkoxy
(linear, branched, optionally substituted); [0971] n is 0-10; and a
dashed line indicates the attachment point to the PTM or ULM
moieties.
[0972] In additional embodiments, the linker (L) comprises a
structure selected from, but not limited to the structure shown
below, where a dashed line indicates the attachment point to the
PTM or ULM moieties:
##STR00444##
wherein: [0973] W.sup.L1 and W.sup.L2 are each independently aryl,
heteroaryl, cyclic, heterocyclic, C.sub.1-6 alkyl, bicyclic,
biaryl, biheteroaryl, or biheterocyclic, each optionally
substituted with R.sup.Q, each R.sup.Q is independently a H, halo,
OH, CN, CF.sub.3, hydroxyl, nitro, C.ident.CH, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.1-C.sub.6 alkyl (linear, branched,
optionally substituted), C.sub.1-C.sub.6 alkoxy (linear, branched,
optionally substituted), OC.sub.1-3alkyl (optionally substituted by
1 or more --F), OH, NH.sub.2, NR.sup.Y1R.sup.Y2, CN, or 2 R.sup.Q
groups taken together with the atom they are attached to, form a
4-8 membered ring system containing 0-4 heteroatoms; [0974]
Y.sup.L1 is each independently a bond, NR.sup.YL1, O, S,
NR.sup.YL2, CR.sup.YL1R.sup.YL2, C.dbd.O, C.dbd.S, SO, SO.sub.2,
C.sub.1-C.sub.6alkyl (linear, branched, optionally substituted) and
optionally one or more C atoms are replaced with O; C.sub.1-C.sub.6
alkoxy (linear, branched, optionally substituted); [0975] Q.sup.L
is a 3-6 membered alicyclic or aromatic ring with 0-4 heteroatoms,
optionally bridged, optionally substituted with 0-6 R.sup.Q, each
R.sup.Q is independently H, C.sub.1-6 alkyl (linear, branched,
optionally substituted by 1 or more halo, C.sub.1-6 alkoxyl), or 2
R.sup.Q groups taken together with the atom they are attached to,
form a 3-8 membered ring system containing 0-2 heteroatoms); [0976]
R.sup.YL1, R.sup.YL2 are each independently H, OH, C.sub.1-6 alkyl
(linear, branched, optionally substituted by 1 or more halo,
C.sub.1-6 alkoxyl), or R.sup.1, R.sup.2 together with the atom they
are attached to, form a 3-8 membered ring system containing 0-2
heteroatoms); [0977] n is 0-10; and a dashed line indicates the
attachment point to the PTM or ULM moieties.
[0978] In additional embodiments, the linker group is optionally
substituted (poly)ethyleneglycol having between 1 and about 100
ethylene glycol units, between about 1 and about 50 ethylene glycol
units, between 1 and about 25 ethylene glycol units, between about
1 and 10 ethylene glycol units, between 1 and about 8 ethylene
glycol units and 1 and 6 ethylene glycol units, between 2 and 4
ethylene glycol units, or optionally substituted alkyl groups
interdispersed with optionally substituted, O, N, S, P or Si atoms.
In certain embodiments, the linker is substituted with an aryl,
phenyl, benzyl, alkyl, alkylene, or heterocycle group. In certain
embodiments, the linker may be asymmetric or symmetrical.
[0979] In any of the embodiments of the compounds described herein,
the linker group may be any suitable moiety as described herein. In
one embodiment, the linker is a substituted or unsubstituted
polyethylene glycol group ranging in size from about 1 to about 12
ethylene glycol units, between 1 and about 10 ethylene glycol
units, about 2 about 6 ethylene glycol units, between about 2 and 5
ethylene glycol units, between about 2 and 4 ethylene glycol
units.
[0980] In another embodiment, the present disclosure is directed to
a compound which comprises a PTM group as described herein, which
binds to a target protein, e.g., EGFR or polypeptide derived
therefrom, which is ubiquitinated by an ubiquitin ligase and is
chemically linked directly to the ULM group or through a linker
moiety L, or PTM is alternatively a ULM' group which is also a
ubiquitin ligase binding moiety, which may be the same or different
than the ULM group as described above and is linked directly to the
ULM group directly or through the linker moiety; and L is a linker
moiety as described above which may be present or absent and which
chemically (covalently) links ULM to PTM, or a pharmaceutically
acceptable salt, enantiomer, stereoisomer, solvate or polymorph
thereof.
[0981] In certain embodiments, the linker group L is a group
comprising one or more covalently connected structural units
independently selected from the group consisting of:
##STR00445##
The X is selected from the group consisting of O, N, S, S(O) and
SO.sub.2; n is integer from 1-5, 5; R.sup.L1 is hydrogen or
alkyl,
##STR00446##
is a mono- or bicyclic aryl or heteroaryl optionally substituted
with 1-3 substituents selected from alkyl, halogen, haloalkyl,
hydroxy, alkoxy or cyano;
##STR00447##
is a mono- or bicyclic cycloalkyl or a heterocycloalkyl optionally
substituted with 1-3 substituents selected from alkyl, halogen,
haloalkyl, hydroxy, alkoxy or cyano; and the phenyl ring fragment
can be optionally substituted with 1, 2 or 3 substituents selected
from the group consisting of alkyl, halogen, haloalkyl, hydroxy,
alkoxy and cyano. In an embodiment, the linker group L comprises up
to 10 covalently connected structural units, as described
above.
[0982] Without being limited by any particular theory, the
inventors believe that the composition and structure of the linker,
although unlimited in principal, can have significant effects on
the efficacy and potency of the bifunctional compound as described
herein; perhaps due to modulation of the interaction between the
ULM and the PTM. However, the linker can be optimized according to
the present teachings without undue experimentation.
[0983] Although the ULM group and PTM group may be covalently
linked to the linker group through any group which is appropriate
and stable to the chemistry of the linker, in preferred aspects of
the present disclosure, the linker is independently covalently
bonded to the ULM group and the PTM group preferably through an
amide, ester, thioester, keto group, carbamate (urethane), carbon
or ether, each of which groups may be inserted anywhere on the ULM
group and PTM group to provide maximum binding of the ULM group on
the ubiquitin ligase and the PTM group on the target protein to be
degraded. (It is noted that in certain aspects where the PTM group
is a ULM group, the target protein for degradation may be the
ubiquitin ligase itself). In certain preferred aspects, the linker
may be linked to an optionally substituted alkyl, alkylene, alkene
or alkyne group, an aryl group or a heterocyclic group on the ULM
and/or PTM groups.
Exemplary PTMs
[0984] In preferred aspects of the disclosure, the PTM group is a
moiety that binds to a target protein of interest. Targets of the
PTM group are numerous in kind and are selected from proteins that
are expressed in a cell such that at least a portion of the
sequences is found in the cell and may bind to a PTM group. The
term "protein" includes oligopeptides and polypeptide sequences of
sufficient length that they can bind to a PTM group according to
the present disclosure. Any protein in a eukaryotic system or a
microbial system, including a virus, bacteria or fungus, as
otherwise described herein, are targets for ubiquitination mediated
by the compounds according to the present disclosure. Preferably,
the target protein is a eukaryotic protein.
[0985] PTM groups according to the present disclosure include, for
example, any moiety which binds to an RTK target protein,
including, by way of non-limiting example, EGFR, HER2, c-MET, IGFR.
The compositions described below exemplify some of the members of
small molecule target protein binding moieties. Such small molecule
target protein binding moieties also include pharmaceutically
acceptable salts, enantiomers, solvates and polymorphs, prodrug and
deuterated forms of the compounds. These binding moieties are
linked to the ubiquitin ligase binding moiety preferably through a
linker as described herein in order to present the RTK (to which
the protein target moiety is bound) in proximity to the ubiquitin
ligase for ubiquitination and degradation.
[0986] The present disclosure may be used to treat a number of
disease states and/or conditions, including any disease state
and/or condition in which RTK proteins are dysregulated, e.g.,
cancer and/or inflammatory disorders, where a patient would benefit
from the degradation of proteins.
[0987] In an additional aspect, the description provides
therapeutic compositions comprising an effective amount of a
compound as described herein or salt form thereof, and a
pharmaceutically acceptable carrier, additive or excipient, and
optionally an additional bioactive agent. The therapeutic
compositions modulate protein degradation in a patient or subject,
for example, an animal such as a human, and can be used for
treating or ameliorating disease states or conditions which are
modulated through the degraded protein. In certain embodiments, the
therapeutic compositions as described herein may be used to
effectuate the degradation of proteins of interest for the
treatment or amelioration of a disease, e.g., cancer. In certain
additional embodiments, the disease is an inflammatory
disorder.
[0988] In alternative aspects, the present disclosure relates to a
method for treating a disease state or ameliorating the symptoms of
a disease or condition in a subject in need thereof by degrading a
protein or polypeptide through which a disease state or condition
is modulated comprising administering to said patient or subject an
effective amount, e.g., a therapeutically effective amount, of at
least one compound as described hereinabove, optionally in
combination with a pharmaceutically acceptable carrier, additive or
excipient, and optionally an additional bioactive agent, wherein
the composition is effective for treating or ameliorating the
disease or disorder or symptom thereof in the subject. The method
according to the present disclosure may be used to treat a large
number of disease states or conditions including cancer and/or an
inflammatory disorder, by virtue of the administration of effective
amounts of at least one compound described herein. The disease
state or condition may be a disease caused by a microbial agent or
other exogenous agent such as a virus, bacteria, fungus, protozoa
or other microbe or may be a disease state, which is caused by
overexpression of a protein, which leads to a disease state and/or
condition.
[0989] In another aspect, the description provides methods for
identifying the effects of the degradation of proteins of interest
in a biological system using compounds according to the present
disclosure.
[0990] As used herein, unless the context indicates otherwise, the
term "target protein" is used to describe a protein or polypeptide,
e.g., an RTK, such as EGFR, HER2, c-MET, IGFR, which is a target
for binding to a compound according to the present disclosure and
degradation by ubiquitin ligase hereunder. Such small molecule
target protein binding moieties also include pharmaceutically
acceptable salts, enantiomers, solvates and polymorphs, prodrugs
and deuterated forms of these compounds, as well as other small
molecules that may target a protein of interest. These binding
moieties are linked to at least one ULM group (e.g. VLM, CLM, ILM,
and/or MLM) through at least one linker group L. In certain
aspects, target proteins, which may be bound to the protein target
moiety and degraded by the ligase to which the ubiquitin ligase
binding moiety is bound, include any protein or peptide, including
fragments thereof, analogues thereof, and/or homologues
thereof.
[0991] Epidermal growth factor receptor (EGFR) exists on the cell
surface and is activated by binding of its specific ligands,
including epidermal growth factor and transforming growth factor
.alpha. (TGF.alpha.). Upon activation by its growth factor ligands,
EGFR undergoes a transition from an inactive monomeric form to an
active homodimer--although there is some evidence that preformed
inactive dimers may also exist before ligand binding. In addition
to forming homodimers after ligand binding, EGFR may pair with
another member of the ErbB receptor family, such as ErbB2/Her2/neu,
to create an activated heterodimer. There is also evidence to
suggest that clusters of activated EGFRs form, although it remains
unclear whether this clustering is important for activation itself
or occurs subsequent to activation of individual dimers.
[0992] EGFR dimerization stimulates its intrinsic intracellular
protein-tyrosine kinase activity. As a result, autophosphorylation
of several tyrosine (Y) residues in the C-terminal domain of EGFR
occurs. These include Y992, Y1045, Y1068, Y1148 and Y1173, as shown
in the adjacent diagram. This autophosphorylation elicits
downstream activation and signaling by several other proteins that
associate with the phosphorylated tyrosines through their own
phosphotyrosine-binding SH2 domains. These downstream signaling
proteins initiate several signal transduction cascades, principally
the MAPK, Akt and JNK pathways, leading to DNA synthesis and cell
proliferation. Such proteins modulate phenotypes such as cell
migration, adhesion, and proliferation. Activation of the receptor
is important for the innate immune response in human skin. The
kinase domain of EGFR can also cross-phosphorylate tyrosine
residues of other receptors it is aggregated with, and can itself
be activated in that manner.
[0993] Mutations that lead to EGFR overexpression (known as
upregulation) or overactivity have been associated with a number of
cancers, including squamous-cell carcinoma of the lung, anal
cancers, glioblastoma, and epithelial tumors of the head and neck.
These somatic mutations involving EGFR lead to its constant
activation, which produces uncontrolled cell division. In
glioblastoma a more or less specific mutation of EGFR, called
EGFRvIII is often observed. Mutations, amplifications or
misregulations of EGFR or family members are implicated in about
30% of all epithelial cancers. Aberrant EGFR signaling has been
implicated in inflammatory disorders, e.g., psoriasis, eczema and
atherosclerosis.
[0994] In certain embodiments, the description provides
compositions and methods for treating an EGFR-related disease or
disorder. In certain embodiments, the EGFR-related disease or
disorder is at least one of squamous-cell carcinoma of the lung,
colon and anal cancers, glioblastoma, and epithelial tumors of the
head and neck, psoriasis, eczema and atherosclerosis or a
combination thereof.
[0995] The identification of EGFR as an oncogene has led to the
development of anticancer therapeutics directed against EGFR
(called "EGFR inhibitors"), including gefitinib, erlotinib,
afatinib, brigatinib and icotinib for lung cancer, and cetuximab
for colon cancer. More recently AstraZeneca has developed
Osimertinib, a third generation tyrosine kinase inhibitor.
[0996] Many therapeutic approaches are aimed at the EGFR. Cetuximab
and panitumumab are examples of monoclonal antibody inhibitors.
However the former is of the IgG1 type, the latter of the IgG2
type. Other monoclonals in clinical development are zalutumumab,
nimotuzumab, and matuzumab. The monoclonal antibodies block the
extracellular ligand binding domain. With the binding site blocked,
signal molecules can no longer attach there and activate the
tyrosine kinase.
[0997] Another method is using small molecules to inhibit the EGFR
tyrosine kinase, which is on the cytoplasmic side of the receptor.
Without kinase activity, EGFR is unable to activate itself, which
is a prerequisite for binding of downstream adaptor proteins. By
halting the signaling cascade in cells that rely on this pathway
for growth, tumor proliferation and migration is diminished or
inhibited. Gefitinib, erlotinib, brigatinib and lapatinib (mixed
EGFR and ERBB2 inhibitor) are examples of small molecule kinase
inhibitors.
[0998] CimaVax-EGF, an active vaccine targeting EGF as the major
ligand of EGFR, uses a different approach, raising antibodies
against EGF itself, thereby denying EGFR-dependent cancers of a
proliferative stimulus; it is in use as a cancer therapy against
non-small-cell lung carcinoma (the most common form of lung
cancer).
[0999] The protein target may be used in screens that identify
compound moieties which bind to the protein and by incorporation of
the moiety into compounds according to the present disclosure, the
level of activity of the protein may be altered for therapeutic end
result.
[1000] The compositions described below exemplify some of the
members of EGFR-binding PTMs that can be incorporated into PROTAC
compounds as described herein. Such small molecule target protein
binding moieties also include pharmaceutically acceptable salts,
enantiomers, solvates and polymorphs, prodrugs and deuterated forms
of these compounds, as well as other small molecules that may
target a protein of interest.
[1001] In certain exemplary embodiments, the PTM (protein-targeting
moiety) of the PROTAC compound as described herein is selected from
EAI045, afatinib, brigatinib, cabozantinib, crizotinib,
dacomitinib, erlotinib, foretinib, gefitinib, icotinib, imatinib,
lapatinib, lenvatinib, motesanib, neratinib, osimertinib,
pazopanib, suntinib, tivantinib, vandetanib, INCB28060, AMG-458,
PF-04217903, PF-02341066, E7050, MK-2461, MBS-777607, JNJ-38877605,
ARQ197, GSK/1363089/XL880, XL184, analogs, derivatives, polymorphs
or solvates thereof.
[1002] In certain exemplary embodiments, the PTM (protein-targeting
moiety) of the PROTAC compound as described herein is represented
by the general formulas I through XVII, and embodiments described.
As described herein, the PTMs are coupled via a linker moiety to a
ULM. It is contemplated that the linker moiety can be conjugated at
any location desired on the PTM. In certain preferred embodiments,
the linker moiety is conjugated to at least one R group of the
structures as shown and described below.
[1003] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula I:
##STR00448##
[1004] wherein Z is selected from N, CH or C--CN;
[1005] X is N-alkyl, amido, NH, or O;
[1006] R.sup.21 is 1 to 3 substituents independently selected from
H, halogen, alkyl, e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6
haloalkyl, alkenyl, e.g., C2-C6 alkenyl, alkynyl, e.g., C2-C6
alkynyl, hydroxy, alkoxy, e.g., C1-C6 alkoxy, amino, alkylamino,
e.g., C1-C6 alkylamino, dialkylamino, cyano, aryl, e.g., C5-C10
aryl, heteroaryl, e.g., C3-C10 heteroaryl comprising 1 or more
heteroatoms selected from O, N, and S, cycloalkyl, e.g., C3-C10
cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl wherein
the heterocycle comprises 1 or more heteroatoms selected from O, N,
and S, aryloxy, e.g., C5-C10 aryloxy, and heteroaryloxy, e.g.,
C5-C10 heteroaryloxy wherein the heteroaryl comprises 1 or more
heteroatoms selected from O, N, and S, arylalkyl, heteroarylalkyl,
arylalkyloxy and heteroarylalkyloxy, wherein the said aryl,
heteroaryl, aryloxy and heteroaryloxy can be further substituted
with 1 to 2 substituents selected from alkyl, e.g., C1-C6 alkyl,
halogen, haloalkyl, e.g., C1-C6 haloalkyl, hydroxyl, alkoxy, e.g.,
C1-C6 alkoxy, amino, alkylamino, dialkylamino and cyano;
[1007] R.sup.22 is 1 to 2 substituents independently selected from
H, halogen, alkyl, e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6
haloalkyl, alkenyl, e.g., C2-C6 alkenyl, alkynyl, e.g., C2-C6
alkynyl, hydroxy, alkoxy, e.g., C1-C6 alkoxy, amino, alkylamino,
e.g., C1-C6 alkylamino, dialkylamino, cyano, aryl, e.g., C5-C10
aryl, heteroaryl, e.g., C3-C10 heteroaryl comprising 1 or more
heteroatoms selected from O, N, and S, cycloalkyl, e.g., C3-C10
cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl wherein
the heterocycle comprising 1 or more heteroatoms selected from O,
N, and S, furanyl, alkyl or alkylamino substituted furanyl or
heterocycloalkyl, wherein the said aryl or heteroaryl can be
further substituted with 1 to 2 substituents selected from alkyl,
e.g., C1-C6 alkyl, halogen, haloalkyl, e.g., C1-C6 haloalkyl,
hydroxyl, alkoxy, e.g., C1-C6 alkoxy, amino, alkylamino,
dialkylamino, cyano or
R.sup.99SO.sub.2(CH.sub.2).sub.sNHCH.sub.2--, where R.sup.99 is an
alkyl, e.g., C1-C6 alkyl, and s is an integer between 0 and 3, or
one or both of the R.sup.22 substituents can be further selected
from --OR.sup.23 or --NHC(O)R.sup.24, where R.sup.23 is selected
from hydroxyalkyl, e.g., C1-C6 hydroxyalkyl, alkoxyalkyl, e.g.,
C1-C6 alkoxyalkyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl,
aryl, e.g., C5-C10 aryl, toluene optionally substituted with an
alkyl or halogen, heteroaryl, e.g., C5-C10 heteroaryl, cycloalkyl,
e.g., C3-C10 cycloalkyl, heterocycloalkyl, C3-C10 heterocycloalkyl
wherein the heterocycle comprising 1 or more heteroatoms selected
from O, N, and S, arylalkyl, e.g., C5-C10 arylalkyl,
heteroarylalkyl, e.g., C5-C10 heteroarylalkyl wherein the
heteroaryl comprises 1 or more heteroatoms selected from O, N, and
S, arylalkyl, e.g., C5-C10 arylalkyl, cycloalkylalkyl, or
heterocycloalkylalkyl as described herein, with the proviso that
the two heteroatoms are not attached to the same carbon atom, and
R.sup.24 is selected from alkyl or from the groups below:
##STR00449##
and
[1008] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl, e.g., C1-C6 alkyl, with the proviso that
R.sup.25 and R.sup.26 taken together with the N atom, to which they
are connected, may form a heterocycloalkyl.
[1009] In certain embodiments, the PTM of formula I comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula I comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1010] In certain additional embodiments, one or more of R.sup.21,
R.sup.22, R.sup.23, R.sup.24, R.sup.25 or R.sup.26 groups of
formula I is coupled to a linker, wherein the linker is a chemical
moiety as described herein coupling the PTM to the ULM group.
[1011] In certain embodiments, formula I has the structure:
##STR00450##
wherein each R.sup.21 is independently selected from H, halogen,
Cl, F, alkyl, e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6 haloalkyl,
alkynyl, e.g., C1-C6 alkynyl, alkoxy, e.g., C1-C6 alkoxy,
arylmethoxy and heteroarylmethoxy, wherein the said aryl and
heteroaryl can be further substituted with 1 to 2 substituents
selected from alkyl, halogen and haloalkyl; and
[1012] each R.sup.22 is independently selected from H, N, halogen,
alkyl, e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6 haloalkyl, alkoxy,
e.g., C1-C6 alkoxy, methoxy, ethoxy, amino, amido, alkylamino,
dialkylamino, cyano, aryl, e.g., C5-C10 aryl, heteroaryl, e.g.,
C5-C10 heteroaryl comprising from 1 or more heteroatoms selected
from N, O and S, furan, pyrrole, imidazole, oxazole, isoxazole,
thazole, cycloalkyl, e.g., C3-C10 cycloalkyl, heterocycloalkyl,
e.g., C3-C10 heterocycloalkyl wherein the heterocycle comprises
from 1 or more heteroatoms selected from N, O, and S, wherein the
said aryl or heteroaryl can be further substituted with 1 to 2
substituents selected from alkyl, e.g., C1-C6 alkyl, halogen,
haloalkyl, cyano or R.sup.99SO2(CH2)sNHCH2-, where R.sup.99 is an
alkyl, e.g., C1-C6 alkyl, and s is an integer between 0 and 3,
--OR.sup.23 or --NHC(O)R.sup.24, where R.sup.23 is selected from
hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, or
dialkylaminoalkyl, with the proviso that the two heteroatoms are
not attached to the same carbon atom, and R.sup.24 is selected from
the groups below:
##STR00451##
and
[1013] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl with the proviso that R.sup.25 and R.sup.26 taken
together with the N atom, to which they are connected, may form a
heterocycloalkyl.
[1014] In certain embodiments, the PTM of formula I comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula I comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1015] In certain additional embodiments, one or more of R.sup.21,
R.sup.22, R.sup.23, R.sup.24, R.sup.25 or R.sup.26 groups of
formula I is coupled to a linker, wherein the linker is a chemical
moiety as described herein coupling the PTM to the ULM group.
[1016] In certain embodiments, formula I has the structure:
##STR00452##
[1017] wherein each R.sup.21 s independently selected from H,
halogen, Cl, F, alkyl, e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6
haloalkyl, alkynyl, e.g., C1-C6 alkynyl, alkoxy, e.g., C1-C6
alkoxy, arylmethoxy, e.g., C5-C10 arylmethoxy, heteroarylmethoxy,
e.g., C5-C10 heteroarylmethoxy wherein the heteroaryl comprises
from 1 or more heteroatoms selected from N, O, and S, or
##STR00453##
wherein the said aryl and heteroaryl can be further substituted
with 1 to 2 substituents selected from alkyl, halogen and
haloalkyl; and
[1018] each R.sup.22 is independently selected from H, N, halogen,
alkyl, e.g., C1-C6 alkyl, haloalkyl, alkoxy, methoxy, amino, amido,
alkylamino, dialkylamino, cyano, aryl, heteroaryl, furan, pyrrole,
imidazole, oxazole, isoxazole, thazole, cycloalkyl,
heterocycloalkyl--wherein the said aryl or heteroaryl can be
further substituted with 1 to 2 substituents selected from alkyl,
halogen, haloalkyl, cyano or R.sup.99SO2(CH2)sNHCH2-, where
R.sup.99 is an alkyl, and s is an integer between 0 and 3,
--OR.sup.23 or --NHC(O)R.sup.24, where R.sup.23 is selected from
hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, or
dialkylaminoalkyl, with the proviso that the two heteroatoms are
not attached to the same carbon atom, and R.sup.24 is selected from
the groups below:
##STR00454##
and
[1019] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl with the proviso that R.sup.25 and R.sup.26 taken
together with the N atom, to which they are connected, may form a
heterocycloalkyl.
[1020] In certain additional embodiments, one or more of R.sup.21,
R.sup.22, R.sup.23, R.sup.24, R.sup.25 or R.sup.26 groups of
formula I is coupled to a linker, wherein the linker is a chemical
moiety as described herein coupling the PTM to the ULM group.
[1021] In certain embodiments, PTMs of the PROTACs as described
herein comprise the moiety represented by formula I as above:
[1022] wherein Z is N or C--CN, most preferably N;
[1023] R.sup.21 is 1 to 2 substituents independently selected from
H, halogen, alkyl, haloalkyl, alkynyl, alkoxy, arylmethyloxy and
heteroarylmethyloxy, wherein the said aryl and heteroaryl can be
further substituted with 1 to 2 substituents selected from alkyl,
halogen and haloalkyl.
[1024] R.sup.22 is 1 or 2 substituents each independently selected
from H, halogen, alkyl, haloalkyl, alkoxy, amino, alkylamino,
dialkylamino, cyano, aryl, heteroaryl, cycloalkyl, heterocycloalkyl
wherein the said aryl or heteroaryl can be further substituted with
1 to 2 substituents selected from alkyl, halogen, haloalkyl, cyano
or R.sup.99SO2(CH2)sNHCH2-, where R.sup.99 is an alkyl, and s is an
integer between 0 and 3--or R22 is 1 to 2 substituents selected
from --OR23 or --NHC(O)R.sup.24, where R.sub.23 is selected from
hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, or
dialkylaminoalkyl, with the proviso that the two heteroatoms are
not attached to the same carbon atom, and R.sup.24 is selected from
the groups below:
##STR00455##
and
[1025] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl with the proviso that R.sup.25 and R.sup.26 taken
together with the N atom, to which they are connected, may form a
heterocycloalkyl.
[1026] In certain embodiments, the PTM of formula I comprises the
structure:
##STR00456##
wherein each R.sup.21 is independently selected from H, a halogen,
Cl, or F; and each R.sup.22 is independently selected from H, O, N,
C1-C6 alkoxy, or methoxy, amine, amido or
##STR00457##
O-linked 5- or 7-membered ring, e.g., ozaxole, isoxazole,
imidazole, pyrrole, pyrrolidinyl, pyrazole, furan, or thiazole, and
wherein at least one R22 group is coupled to a linker group,
wherein linker is a chemical moiety coupling the PTM to a ULM
group.
[1027] In certain additional embodiments, one or more of R.sup.21,
R.sup.22, R.sup.23, R.sup.24, R.sup.25 or R.sup.26 groups of
formula I is coupled to a linker, wherein the linker is a chemical
moiety as described herein coupling the PTM to the ULM group.
[1028] In certain embodiments, the PTM of formula I has the
structure:
##STR00458##
wherein the linker (L) is a chemical moiety coupling the PTM to a
ULM group.
[1029] In certain embodiments, the PTM of formula I comprises the
structure selected from:
##STR00459##
wherein linker (L) is a chemical moiety coupling the PTM to a ULM
group.
[1030] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula II:
##STR00460##
[1031] wherein R.sup.21 is 1 to 3 substituents independently
selected from H, halogen, Cl, F, alkyl, e.g., C1-C6 alkyl,
haloalkyl, e.g., C1-C6 haloalkyl, alkenyl, e.g., C2-C6 alkenyl,
alkynyl, e.g., C2-C6 alkynyl, hydroxy, alkoxy, e.g., C1-C6 alkoxy,
amino, alkylamino, dialkylamino, cyano, aryl, heteroaryl, e.g.,
C3-C10 heteroaryl comprising 1 or more heteroatoms selected from O,
N, and S, cycloalkyl, e.g., C3-C10 cycloalkyl, heterocycloalkyl,
e.g., C3-C10 heterocycloalkyl wherein the heterocycle comprising 1
or more heteroatoms selected from O, N, and S, aryloxy and
heteroaryloxy, arylalkyl, arylalkyl, e.g., C5-C10 arylalkyl,
heteroarylalkyl, heteroarylalkyl, e.g., C5-C10 heteroarylalkyl
wherein the heteroaryl comprises 1 or more heteroatoms selected
from O, N, and S, arylalkyloxy and heteroarylalkyloxy, wherein the
said aryl, heteroaryl, aryloxy and heteroaryloxy can be further
substituted with 1 to 2 substituents selected from alkyl, e.g.,
C1-C6 alkyl, halogen, haloalkyl, haloalkyl, e.g., C1-C6 haloalkyl,
hydroxyl, alkoxy, alkoxy, e.g., C1-C6 alkoxy, amino, alkylamino,
dialkylamino and cyano;
[1032] R.sup.27 is selected from H, alkyl, e.g., C1-C6 alkyl,
hydroxyalkyl and alkoxyalkyl;
[1033] A is an aryl, aryl, e.g., C5-C10 aryl, a phenyl, or a
heteroaryl, e.g., C3-C10 heteroaryl comprising 1 or more
heteroatoms selected from O, N, and S; and
[1034] R.sup.28 is 1 to 2 substituents independently selected from
H, O, N, halogen, alkyl, e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6
haloalkyl, alkenyl, e.g., C2-C6 alkenyl, alkynyl, e.g., C2-C6
alkynyl, hydroxy, alkoxy, e.g., C1-C6 alkoxy, alkylamino, e.g.,
C1-C6 alkylamino, dialkylamino, cyano, aryl, aryl, e.g., C5-C10
aryl, heteroaryl, e.g., C3-C10 heteroaryl comprising 1 or more
heteroatoms selected from O, N, and S, cycloalkyl, e.g., C3-C10
cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl wherein
the heterocycle comprising 1 or more heteroatoms selected from O,
N, and S, wherein the said aryl or heteroaryl can be further
substituted with 1 to 2 substituents selected from alkyl, e.g.,
C1-C6 alkyl, halogen, haloalkyl, e.g., C1-C6 haloalkyl, hydroxyl,
alkoxy, e.g., C1-C6 alkoxy, amino, alkylamino, dialkylamino and
cyano.
[1035] In certain embodiments, the PTM of formula II comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula II comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1036] In certain additional embodiments, one or more of R.sup.21,
R.sup.27 or R.sup.28 groups of formula II is coupled to a linker,
wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1037] In certain embodiments, PTMs of the PROTACs as described
herein comprise the moiety represented by the formula II as
above:
[1038] wherein R.sup.21 is 1 to 2 substituents independently
selected from H, halogen, Cl, F, alkyl, e.g., C1-C6 alkyl,
haloalkyl, e.g., C1-C6 haloalkyl, alkynyl, e.g., C1-C6 alkynyl,
alkoxy, e.g., C1-C6 alkoxy, arylmethoxy, e.g., a 5-7 membered ring
arylmethoxy, cyano, and heteroarylmethoxy, wherein the said aryl
and heteroaryl can be further substituted with 1 to 2 substituents
selected from alkyl, e.g., C1-C6 alkyl, halogen and haloalkyl,
e.g., C1-C6 haloalkyl;
[1039] R.sup.27 is selected from H, methyl, ethyl, hydroxymethyl
and methoxymethyl;
[1040] A is phenyl or a pyridyl, most preferably phenyl; and
[1041] R.sup.28 is 1 to 2 substituents independently selected from
H, O, N, halogen, alkyl, e.g., C.sub.1-C6 alkyl, haloalkyl, e.g.,
C1-C6 haloalkyl, and cyano.
[1042] In certain embodiments of formula II, the PTM has the
structure:
##STR00461##
[1043] wherein R.sup.21 is H, halogen, Cl, F, alkyl, e.g., C1-C6
alkyl, haloalkyl, alkynyl, alkoxy, arylmethyloxy, cyano, and
heteroarylmethyloxy, wherein the said aryl and heteroaryl can be
further substituted with 1 to 2 substituents selected from alkyl,
halogen and haloalkyl;
[1044] R.sup.27 is selected from H, methyl, ethyl, hydroxymethyl
and methoxymethyl; R.sup.28 is selected from H, O, N, halogen,
C1-C6 alkyl, haloalkyl, e.g., C1-C6 haloalkyl, alkenyl, e.g., C2-C6
alkenyl, alkynyl, e.g., C2-C6 alkynyl, alkoxy, e.g., C1-C6 alkoxy,
alkylamino, e.g., C1-C6 alkylamino, dialkylamino, cyano, aryl,
e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10 heteroaryl comprising 1
or more heteroatoms selected from O, N, and S, cycloalkyl, e.g.,
C3-C10 cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl
wherein the heterocycle comprising 1 or more heteroatoms selected
from O, N, and S, wherein the said aryl or heteroaryl can be
further substituted with 1 to 2 substituents selected from alkyl,
e.g., C1-C6 alkyl, halogen, haloalkyl, hydroxyl, alkoxy, amino,
alkylamino, dialkylamino and cyano as defined above.
[1045] In certain embodiments, R.sup.28 is coupled to a linker,
wherein the linker is a chemical moiety coupling the PTM to the ULM
group.
[1046] In certain additional embodiments, one or more of R.sup.21,
R.sup.27 or R.sup.28 groups of formula II is coupled to a linker,
wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1047] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula III:
##STR00462##
[1048] wherein R.sup.21 is as described above;
[1049] R.sup.29 is 1 to 2 substituents independently selected from
H, halogen, aryl alkyl, haloalkyl, alkoxy and cyano; and
[1050] R.sup.30 is selected from H, hydroxyalkyl, e.g., C1-C6
hydroxyalkyl, alkoxyalkyl, e.g., C1-C6 alkoxyalkyl, aminoalkyl,
alkylaminoalkyl, dialkylaminoalkyl, aryl, e.g., C5-C10 aryl,
heteroaryl, e.g., C3-C10 heteroaryl comprising 1 or more
heteroatoms selected from O, N, and S, cycloalkyl, e.g., C3-C10
cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl wherein
the heterocycle comprising 1 or more heteroatoms selected from O,
N, and S, arylalkyl, e.g., C5-C10 arylalkyl, heteroarylalkyl, e.g.,
C5-C10 heteroarylalkyl wherein the heteroaryl comprises 1 or more
heteroatoms selected from O, N, and S, cycloalkylalkyl, or
heterocycloalkylalkyl, wherein the said alkyl could be optionally
interrupted by --NHC(O)-- or --C(O)NH-- groups, with the proviso
that two heteroatoms are not attached to the same carbon atom.
[1051] In certain embodiments, the PTM of formula III, comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula III comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1052] In certain additional embodiments, one or more of R.sup.21,
R.sup.29 or R.sup.30 groups of formula III is coupled to a linker,
wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1053] In certain embodiments, the PTMs of the PROTACs as described
herein comprise the moiety represented by the formula III:
[1054] wherein R.sup.21 is 1 to 2 substituents independently
selected from H, halogen, alkyl, e.g., C1-C6 alkyl, haloalkyl,
e.g., C1-C6 haloalkyl, alkynyl, e.g., C1-C6 alkynyl, alkoxy, e.g.,
C1-C6 alkoxy, arylmethyloxy and heteroarylmethyloxy as described
herein, wherein the said aryl and heteroaryl can be further
substituted with 1 to 2 substituents selected from alkyl, e.g.,
C1-C6 alkyl, halogen and haloalkyl, e.g., C1-C6 haloalkyl;
[1055] R.sup.29 is 1 substituent selected from H, halogen, alkyl,
e.g., C1-C6 alkyl, or haloalkyl, e.g., C1-C6 haloalkyl; and
[1056] R.sup.30 is selected from H, hydroxyalkyl, e.g., C1-C6
hydroxyalkyl, alkoxyalkyl, e.g., C1-C6 alkoxyalkyl, aminoalkyl,
alkylaminoalkyl, dialkylaminoalkyl, wherein the said alkyl could be
optionally interrupted by --NHC(O)-- or --C(O)NH-- groups, with the
proviso that two heteroatoms are not attached to the same carbon
atom.
[1057] In certain embodiments of formula III, the PTM has the
structure:
##STR00463##
[1058] wherein each R.sup.21 is independently H, halogen, Cl, F,
alkyl, e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6 haloalkyl,
alkenyl, e.g., C2-C6 alkenyl, alkynyl, e.g., C2-C6 alkynyl, alkoxy,
e.g., C1-C6 alkoxy, arylmethyloxy, cyano, and heteroarylmethyloxy,
wherein the said aryl and heteroaryl can be further substituted
with 1 to 2 substituents selected from alkyl, e.g., C1-C6 alkyl,
halogen, or haloalkyl, e.g., C1-C6 haloalkyl;
[1059] Z is C or N; and
[1060] R.sup.28 is defined as above.
[1061] In certain additional embodiments, one or more of R.sup.21
or R.sup.28 groups of formula III is coupled to a linker, wherein
the linker is a chemical moiety as described herein coupling the
PTM to the ULM group.
[1062] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula IV:
##STR00464##
[1063] wherein R.sup.31 is selected from H, halogen, Cl, alkyl,
e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6, haloalkyl, CF.sub.3,
alkoxy, e.g., C1-C6 alkoxy, hydroxyl, cyano, amino, alkylamino,
dialkylamino, aryl, e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10
heteroaryl comprising 1 or more heteroatoms selected from O, N, and
S, indole, pyrazole, imidazole, cycloalkyl, cycloalkyl, e.g.,
C3-C10 cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl
wherein the heterocycle comprising 1 or more heteroatoms selected
from O, N, and S, wherein the said aryl and heteroaryl can be
further substituted with 1 to 2 substituents selected from alkyl,
e.g., C1-C6 alkyl, halogen, haloalkyl, e.g., C1-C6 haloalkyl,
hydroxyl, alkoxy, e.g., C1-C6 alkoxy, or cyano;
[1064] R.sup.32 is selected from H, a 5- or 6-membered aryl or
heteroaryl, a bicyclic fused aryl or heteroaryl and a bicyclic
fused aryl or heteroaryl additionally optionally fused to a
cycloalkyl, e.g., C3-C10 cycloalkyl, or a heterocycloalkyl, e.g.,
C3-C10 heterocycloalkyl wherein the heterocycle comprising 1 or
more heteroatoms selected from O, N, and S, wherein the said aryl
or heteroaryl is optionally substituted with 1-2 substituents
selected from alkyl, e.g., C1-C6 alkyl, halogen, or haloalkyl,
e.g., C1-C6 haloalkyl;
[1065] R.sup.33 is selected from H, alkyl, haloalkyl, halogen,
alkoxy, hydroxyl, amino, alkylamino, dialkylamino and cyano;
[1066] R.sup.34 is selected from H, O, alkyl, e.g., C1-C6 alkyl,
haloalkyl, e.g., C1-C6 haloalkyl, halogen, amino, amido, alkyne,
e.g., C2-C6 alkyne, alkoxy, cyano or --NHC(O)R.sup.24, where
R.sup.24 is R.sup.24 is selected from the groups below:
##STR00465##
and
[1067] R.sup.25 and R.sup.26 are independently selected from H, N,
N(CH.sub.2).sub.1-3, or alkyl, C1-C6 alkyl, with the proviso that
R.sup.25 and R.sup.26 taken together with the N atom, to which they
are connected, may form a heterocycloalkyl; and
[1068] R.sup.35 is selected from --OR.sup.36 and
--NR.sup.37R.sup.38, wherein R.sup.36 is selected from
hydroxyalkyl, e.g., C1-C6 hydroxyalkyl, alkoxyalkyl, e.g., C1-C6
alkoxyalkyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, aryl,
e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10 heteroaryl comprising 1
or more heteroatoms selected from O, N, and S, cycloalkyl, e.g.,
C3-C10 cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl
wherein the heterocycle comprising 1 or more heteroatoms selected
from O, N, and S, arylalkyl, e.g., C5-C10 arylalkyl,
heteroarylalkyl, e.g., C5-C10 heteroarylalkyl wherein the
heteroaryl comprises 1 or more heteroatoms selected from O, N, and
S, cycloalkylalkyl or heterocycloalkylalkyl with the proviso that
the two heteroatoms are not attached to the same carbon atom, and
R.sup.37 and R.sup.38 are independently selected from H or alkyl,
e.g., C1-C6 alkyl, --NR.sup.37R.sup.38, or --NR.sup.37R.sup.38
taken together represent a heterocycloalkyl ring, wherein R.sup.35,
R.sup.37 or R.sup.38 further optionally substituted with alkyl,
e.g., C1-C6 alkyl, hydroxyl, alkoxy, e.g., C1-C6 alkoxy, amino,
alkylamino, dialkylamino, heterocycloalkyl, e.g., C3-C10
heterocycloalkyl wherein the heterocycle comprising 1 or more
heteroatoms selected from O, N, and S, aminoalkyl, alkylaminoalkyl,
dialkylaminoalkyl, heterocycloalkylalkyl or --C(O)R.sup.39 where
R.sup.39 is an alkyl, e.g., C1-C6 alkyl.
[1069] In certain embodiments, the PTM of formula IV comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula IV is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1070] In certain additional embodiments, one or more of R.sup.24,
R.sup.25, R.sup.26, R.sup.31, R.sup.32, R.sup.33, R.sup.34,
R.sup.35, R.sup.36, R.sup.37, R.sup.38 or R.sup.39 groups of
formula IV is coupled to a linker, wherein the linker is a chemical
moiety as described herein coupling the PTM to the ULM group.
[1071] In certain embodiments, R.sup.32 is selected from:
##STR00466##
In certain embodiment, the R.sup.32 group comprises a linker,
wherein the linker is a chemical moiety coupling the PTM to the ULM
group. For example, in certain embodiments, R.sup.32 is:
##STR00467##
[1072] In certain embodiments, R.sup.33 is selected from C1-C6
alkyoxy.
[1073] In certain embodiments, R.sup.35 is selected from:
##STR00468##
In certain embodiments, R.sup.35 group is coupled to a linker,
wherein the linker is a chemical moiety coupling the PTM to the ULM
group.
[1074] In certain embodiments, the PTMs of the PROTACs as described
herein comprise the moiety represented by the formula IV above:
[1075] wherein R.sup.31 is selected from H, halogen, alkyl, e.g.,
C1-C6 alkyl, haloalkyl, e.g., C1-C6 haloalkyl, 5- or 6-membered
heteroaryl, wherein the said heteroaryl can be further substituted
with 1 substituent selected from alkyl, e.g., C1-C6 alkyl, halogen
or haloalkyl, e.g., C1-C6 haloalkyl;
[1076] R.sup.32 is selected from H, a 5- or 6-membered aryl or
heteroaryl, a bicyclic fused aryl or heteroaryl, e.g., each ring
has 4- to 6-members, or a bicyclic fused aryl or heteroaryl
additionally fused to a cycloalkyl or a heterocycloalkyl wherein
the said aryl or heteroaryl is optionally substituted with 1-2
substituents selected from alkyl, e.g., C1-C6 alkyl, halogen, or
haloalkyl, e.g., C1-C6 haloalkyl;
[1077] R.sup.33 is selected from methoxy or ethoxy;
[1078] R.sup.34 is selected from H or --NHC(O)R.sup.24, where
R.sup.24 is selected from the groups below:
##STR00469##
and
[1079] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl, e.g., C1-C6 alkyl, with the proviso that
R.sup.25 and R.sup.26 taken together with the N atom, to which they
are connected, may form a heterocycloalkyl;
[1080] R.sup.35 is --NR.sup.37R.sup.38 where R.sup.37 and R.sup.38
are independently selected from H or alkyl, e.g., C1-C6 alkyl, or
--NR.sup.37R.sup.38 taken together represent a heterocycloalkyl
ring, most preferably piperazine, further optionally substituted
with alkyl, e.g., C1-C6 alkyl, amino, alkylamino, dialkylamino,
aminoalkyl, or --C(O)R.sup.39 where R.sup.39 is an alkyl, e.g.,
C1-C6 alkyl.
[1081] In a further embodiment, R.sup.32 of formula IV is an indole
or an indole further fused to a cyclohexane or a piperidine ring
through the positions 1 and 7 of the indole ring with the proviso
that N atoms are not attached to the same C atom.
[1082] In certain embodiments, PTM of formula IV has the
structure:
##STR00470##
[1083] wherein R.sup.37 and R.sup.38 are as described.
[1084] In certain additional embodiments, one or more of R.sup.37
or R.sup.38 groups of formula IV is coupled to a linker, wherein
the linker is a chemical moiety as described herein coupling the
PTM to the ULM group.
[1085] In certain embodiments, PTM of formula IV has the
structure:
##STR00471##
[1086] In certain embodiments, PTM of formula IV has the structure
selected from the group:
##STR00472##
wherein Linker (L) is a chemical moiety coupling the PTM to a ULM
group.
[1087] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula V:
##STR00473##
[1088] wherein R.sup.31, R.sup.33 and R.sup.35 are as described
above;
[1089] B is selected from an aryl, e.g., C5-C10 aryl, heteroaryl,
e.g., C3-C10 heteroaryl comprising 1 or more heteroatoms selected
from O, N, and S, cycloalkyl, e.g., C3-C10 cycloalkyl,
heterocycloalkyl, e.g., C3-C10 heterocycloalkyl wherein the
heterocycle comprising 1 or more heteroatoms selected from O, N,
and S, provided that in the latter case the heteroatom of the said
heterocycloalkyl is separated from X by at least two carbon
atoms;
[1090] X is O, S or NH; and
[1091] R.sup.40 is --NHC(O)R.sup.24, or --C(O)R.sup.24 if attached
to a ring nitrogen of B where R.sup.24 is as described above.
[1092] In certain embodiments, PTMs of the PROTACs as described
herein comprise the moiety represented by the formula V above:
[1093] wherein R.sup.31 is selected from H, halogen, alkyl, e.g.,
C1-C6 alkyl, haloalkyl, e.g., C1-C6 halolkyl, or 5- or 6-membered
heteroaryl, wherein the said heteroaryl can be further substituted
with 1 substituent selected from alkyl, e.g., C1-C6 alkyl, halogen,
or haloalkyl, e.g., C1-C6 haloalkyl;
[1094] R.sup.33 is selected from methoxy or ethoxy;
[1095] R.sup.35 is --NR.sup.37R.sup.38 where R.sup.37 and R.sup.38
are independently selected from H or alkyl, e.g., C1-C6 alkyl, or
--NR.sup.37R.sup.38 taken together represent a heterocycloalkyl
ring, most preferably piperazine, further optionally substituted
with alkyl, e.g., C1-C6 alkyl, amino, alkylamino, dialkylamino,
aminoalkyl, or --C(O)R.sup.39 where R.sup.39 is an alkyl, e.g.,
C1-C6 alkyl;
[1096] B is phenyl or a pyridyl, most preferably phenyl;
[1097] X is O or NH, most preferably NH;
[1098] R.sup.40 is --NHC(O)R.sup.24, where R.sup.24 is selected
from the groups below:
##STR00474##
and
[1099] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl, e.g., C1-C6 alkyl, with the proviso that
R.sup.25 and R.sup.26 taken together with the N atom, to which they
are connected, may form a heterocycloalkyl.
[1100] In certain embodiments, the PTM of formula V comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula V is coupled to
a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1101] In certain additional embodiments, one or more of R.sup.24,
R.sup.25, R.sup.26, R.sup.31, R.sup.33, R.sup.35, R.sup.37,
R.sup.38, R.sup.39 or R.sup.40 groups of formula V is coupled to a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1102] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula VI:
##STR00475##
[1103] R.sup.41 is selected from H, alkyl, e.g., C1-C6 alkyl,
halogen, or haloalkyl, e.g., C1-C6 haloalkyl, alkoxy, e.g., C1-C6
alkoxy, and cyano;
[1104] R.sup.42 is 1 to 2 substituents independently selected from
H, halogen, alkyl, e.g., C1-C6 alkyl, alkoxy, e.g., C1-C6 alkoxy,
haloalkyl, e.g., C1-C6 haloalkyl, alkenyl, e.g., C2-C6 alkenyl,
alkynyl, e.g., C2-C6 alkynyl, hydroxy, alkoxy, e.g., C1-C6 alkoxy,
amino, alkylamino, dialkylamino, cyano, aryl, e.g., C5-C10 aryl,
heteroaryl, e.g., C3-C10 heteroaryl comprising 1 or more
heteroatoms selected from O, N, and S, imidazole, pyrazole,
pyrrole, cycloalkyl, e.g., C3-C10 cycloalkyl, heterocycloalkyl,
e.g., C3-C10 heterocycloalkyl wherein the heterocycle comprising 1
or more heteroatoms selected from O, N, and S, arylalkyl, e.g.,
C5-C10 arylalkyl, heteroarylalkyl, e.g., C5-C10 heteroarylalkyl
wherein the heteroaryl comprises 1 or more heteroatoms selected
from O, N, and S, cycloalkylalkyl, heterocycloalkylalkyl--wherein
the said aryl or heteroaryl can be further substituted with 1 to 2
substituents selected from alkyl, e.g., C1-C6 alkyl, halogen,
haloalkyl, e.g., C1-C6 haloalkyl, hydroxyl, alkoxy, e.g., C1-C6
alkoxy, amino, alkylamino, dialkylamino and cyano;
[1105] C is an aryl or a heteroaryl, e.g., C3-C10 heteroaryl
comprising 1 or more heteroatoms selected from O, N, and S;
[1106] D is an aryl, e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10
heteroaryl comprising 1 or more heteroatoms selected from O, N, and
S, cycloalkyl or heterocycloalkyl, e.g., 3-8 membered aryl or
heteroaryl comprising at least one heteroatom selected from S, O,
or N;
[1107] Y is a bond, O, S, or NH;
[1108] n is selected from 0, 1 or 2;
[1109] R.sup.43 is 1 to 2 substituents independently selected from
H, halogen, alkyl, e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6
haloalkyl, alkenyl, e.g., C2-C6 alkenyl, alkynyl, e.g., C2-C6
alkynyl, hydroxy, alkoxy, e.g., C1-C6 alkyoxy, amino, alkylamino,
dialkylamino, cyano, aryl, e.g., C5-C10 aryl, heteroaryl, e.g.,
C3-C10 heteroaryl comprising 1 or more heteroatoms selected from O,
N, and S, cycloalkyl, e.g., C3-C10 cycloalkyl, heterocycloalkyl,
e.g., C3-C10 heterocycloalkyl wherein the heterocycle comprising 1
or more heteroatoms selected from O, N, and S, wherein the said
aryl or heteroaryl can be further substituted with 1 to 2
substituents selected from alkyl, e.g., C1-C6 alkyl, halogen,
haloalkyl, haloalkyl, e.g., C1-C6 haloalkyl, hydroxyl, alkoxy,
e.g., C1-C6 alkoxy, amino, alkylamino, dialkylamino and cyano--or
one of the R.sup.43 substituents can be --NHC(O)R.sup.24, or
--C(O)R.sup.24 if attached to a ring nitrogen of D; and
[1110] R.sup.24 is as described above.
[1111] In certain embodiments, the PTM of formula VI comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula VI is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1112] In certain additional embodiments, one or more of R.sup.24,
R.sup.25, R.sup.26, R.sup.41, R.sup.42 or R43 groups of formula VI
is coupled to a linker, wherein the linker is a chemical moiety as
described herein coupling the PTM to the ULM group.
[1113] In certain embodiments, PTMs of the PROTACs as described
herein comprise the moiety represented by the formula VI:
[1114] wherein R.sup.41 is selected from H, halogen, or alkyl,
e.g., C1-C6 alkyl;
[1115] R.sup.42 is 1 substituent independently selected from H,
halogen, alkyl, e.g., C1-C6 alkyl, or haloalkyl, e.g., C1-C6
haloalkyl;
[1116] C is a 5- or 6-membered heteroaryl, most preferably
5-membered heteroaryl;
[1117] D is a heterocycloalkyl, most preferably pyrrolidine;
[1118] Y is a bond, O, or NH, most preferably O;
[1119] n is 1 or 2;
[1120] R.sup.43 is 1 to 2 substituents where one substituent is
--NHC(O)R.sup.24, or --C(O)R.sup.24 if attached to a ring nitrogen
of D, and the other is optionally selected from H, halogen, alkyl,
e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6 haloalkyl, or alkoxy,
e.g., C1-C6 alkoxy;
[1121] R.sup.24 is selected from the groups below:
##STR00476##
and
[1122] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl with the proviso that R.sup.25 and R.sup.26 taken
together with the N atom, to which they are connected, may form a
heterocycloalkyl.
[1123] In certain embodiments, at least one R group comprises a
linker, wherein the linker is a chemical moiety coupling the PTM to
the ULM group.
[1124] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula VII:
##STR00477##
[1125] wherein R.sup.42 is 1 to 2 substituents independently
selected from H, halogen, alkyl, e.g., C1-C6 alkyl, haloalkyl,
e.g., C1-C6 haloalkyl, alkenyl, e.g., C2-C6 alkenyl, alkynyl, e.g.,
C2-C6 alkynyl, hydroxy, alkoxy, e.g., C1-C6 alkoxy, amino,
alkylamino, dialkylamino, cyano, aryl, e.g., C5-C10 aryl,
heteroaryl, e.g., C3-C10 heteroaryl comprising 1 or more
heteroatoms selected from O, N, and S, cycloalkyl, e.g., C3-C10
cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl wherein
the heterocycle comprising 1 or more heteroatoms selected from O,
N, and S, arylalkyl, e.g., C5-C10 arylalkyl, heteroarylalkyl, e.g.,
C5-C10 heteroarylalkyl wherein the heteroaryl comprises 1 or more
heteroatoms selected from O, N, and S, cycloalkylalkyl,
heterocycloalkylalkyl--wherein the said aryl or heteroaryl can be
further substituted with 1 to 2 substituents selected from alkyl,
e.g., C1-C6 alkyl, halogen, haloalkyl, e.g., C1-C6 haloalkyl,
hydroxyl, alkoxy, e.g., C1-C6 alkoxy, amino, alkylamino,
dialkylamino and cyano;
[1126] C is an aryl or a heteroaryl, e.g., imidazole, pyrazole,
pyrrole;
[1127] D is an aryl, heteroaryl, cycloalkyl or heterocycloalkyl,
e.g., pyrrolidinyl;
[1128] Y is a bond, O, S, or NH;
[1129] n is selected from 0, 1 or 2;
[1130] R.sup.43 is 1 to 2 substituents independently selected from
H, halogen, Cl, F, alkyl, e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6
haloalkyl, alkenyl, e.g., C2-C6 alkenyl, alkynyl, e.g., C2-C6
alkynyl, alkoxy, e.g., C1-C6 alkoxy, amino, amido, alkylamino,
e.g., C1-C6 alkylamino, dialkylamino, cyano, aryl, e.g., C5-C10
aryl, heteroaryl, e.g., C3-C10 heteroaryl comprising 1 or more
heteroatoms selected from O, N, and S, cycloalkyl, e.g., C3-C10
cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl wherein
the heterocycle comprising 1 or more heteroatoms selected from O,
N, and S, wherein the said aryl or heteroaryl can be further
substituted with 1 to 2 substituents selected from alkyl, e.g.,
C1-C6 alkyl, halogen, haloalkyl, e.g., C1-C6 haloalkyl, hydroxyl,
alkoxy, e.g., C1-C6 alkoxy, amino, alkylamino, dialkylamino and
cyano, or one of the R.sup.43 substituents can be --NHC(O)R.sup.24,
or --C(O)R.sup.24 if attached to a ring nitrogen of D;
[1131] R.sup.24 is as described above; and
[1132] R.sup.88 is a hydrogen or alkyl, e.g., C1-C6 alkyl.
[1133] In certain embodiments, PTMs of the PROTACs as described
herein comprise the moiety represented by the formula VII:
[1134] wherein R.sup.42 is 1 to 2 substituents independently
selected from H, halogen, alkyl, e.g., C1-C6 alkyl, haloalkyl,
e.g., C1-C6 haloalkyl, and alkoxy, e.g., C1-C6 alkoxy;
[1135] C is a 5- or 6-membered heteroaryl, most preferably
5-membered heteroaryl;
[1136] D is a heterocycloalkyl, most preferably pyrrolidine;
[1137] Y is a bond;
[1138] n is 0;
[1139] R.sup.43 is 1 to 2 substituents where one substituent is
--NHC(O)R.sup.24, or --C(O)R.sup.24 if attached to a ring nitrogen
of D, and the other is optionally selected from H, halogen, alkyl,
e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6 haloalkyl, alkoxy, e.g.,
C1-C6 alkoxy;
[1140] R.sup.24 is selected from the groups below:
##STR00478##
[1141] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl with the proviso that R.sup.25 and R.sup.26 taken
together with the N atom, to which they are connected, may form a
heterocycloalkyl; and
[1142] R.sup.88 is methyl or ethyl.
[1143] In certain embodiments, the PTM of formula VII comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula VII is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1144] In certain additional embodiments, one or more of R.sup.24,
R.sup.25, R.sup.26, R.sup.42, R.sup.43 or R.sup.88 groups of
formula VII is coupled to a linker, wherein the linker is a
chemical moiety as described herein coupling the PTM to the ULM
group.
[1145] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula VIII:
##STR00479##
[1146] wherein R.sup.33, R.sup.35, R.sup.40 and B are as described
above; and
[1147] R.sup.46 is selected from H, alkyl, e.g., C1-C6 alkyl,
hydroxyalkyl, e.g., C1-C6 hydroxyalkyl, alkoxyalkyl, aminoalkyl,
alkylaminoalkyl, dialkylaminoalkyl, aryl, e.g., C5-C10 aryl,
heteroaryl, e.g., C3-C10 heteroaryl comprising 1 or more
heteroatoms selected from O, N, and S, cycloalkyl, e.g., C3-C10
cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl wherein
the heterocycle comprising 1 or more heteroatoms selected from O,
N, and S, arylalkyl, e.g., C5-C10 arylalkyl, heteroarylalkyl, e.g.,
C5-C10 heteroarylalkyl wherein the heteroaryl comprises 1 or more
heteroatoms selected from O, N, and S, cycloalkylalkyl or
heterocycloalkylalkyl with the proviso that the two heteroatoms are
not attached to the same carbon atom and wherein the said aryl and
heteroaryl can be further substituted with 1-2 substituents
selected from alkyl, e.g., C1-C6 alkyl, halogen, alkoxy, e.g.,
C1-C6 alkoxy, and cyano.
[1148] In certain embodiments, PTMs of the PROTACs as described
herein comprise the moiety represented by the formula VIII:
[1149] wherein R.sup.33 is selected from methoxy or ethoxy;
[1150] R.sup.35 is --NR.sup.37R.sup.38 where R.sup.37 and R.sup.38
are independently selected from H or alkyl, e.g., C1-C6 alkyl, or
--NR.sup.37R.sup.38 taken together represent a heterocycloalkyl
ring, most preferably piperazine, further optionally substituted
with alkyl, e.g., C1-C6 alkyl, amino, alkylamino, dialkylamino,
aminoalkyl, or --C(O)R.sup.39 where R.sup.39 is an alkyl, e.g.,
C1-C6 alkyl;
[1151] B is phenyl or a pyridyl, most preferably phenyl;
[1152] R.sup.40 is --NHC(O)R.sup.24, where R.sup.24 is selected
from the groups below:
##STR00480##
[1153] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl with the proviso that R.sup.25 and R.sup.26 taken
together with the N atom, to which they are connected, may form a
heterocycloalkyl; and
[1154] R.sup.46 is selected from H, alkyl, e.g., C1-C6 alkyl,
alkoxyalkyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, aryl,
e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10 heteroaryl comprising 1
or more heteroatoms selected from O, N, and S, arylalkyl, e.g.,
C5-C10 arylalkyl, heteroarylalkyl, e.g., C5-C10 heteroarylalkyl
wherein the heteroaryl comprises 1 or more heteroatoms selected
from O, N, and S, with the proviso that the two heteroatoms are not
attached to the same carbon atom and wherein the said aryl and
heteroaryl can be further substituted with 1-2 substituents
selected from alkyl, e.g., C1-C6 alkyl, and halogen.
[1155] In certain embodiments, the PTM of formula VIII comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula VIII is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1156] In certain additional embodiments, one or more of
[1157] R.sup.24, R.sup.25, R.sup.26, R.sup.33, R.sup.35, R.sup.37,
R.sup.38, R.sup.39, R.sup.40 or R.sup.46 groups of formula VIII is
coupled to a linker, wherein the linker is a chemical moiety as
described herein coupling the PTM to the ULM group.
[1158] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula IX:
##STR00481##
[1159] wherein R.sup.33, R.sup.35, R.sup.40 and B are as described
above; and
[1160] R.sup.47 is selected from H or alkyl, e.g., C1-C6 alkyl.
[1161] In certain embodiments, at least one R group comprises a
linker, wherein the linker is a chemical moiety coupling the PTM to
the ULM group.
[1162] In certain embodiments, PTMs of the PROTACs as described
herein comprise the moiety represented by the formula IX:
[1163] wherein R.sup.33 is selected from methoxy or ethoxy;
[1164] R.sup.35 is --NR.sup.37R.sup.38 where R.sup.37 and R.sup.38
are independently selected from H or alkyl, e.g., C1-C6 alkyl, or
--NR.sup.37R.sup.38 taken together represent a heterocycloalkyl
ring, most preferably piperazine, further optionally substituted
with alkyl, e.g., C1-C6 alkyl, amino, alkylamino, dialkylamino,
aminoalkyl, or --C(O)R.sup.39 where R.sup.39 is an alkyl, e.g.,
C1-C6 alkyl;
[1165] B is phenyl or a pyridyl, most preferably phenyl;
[1166] R.sup.40 is --NHC(O)R.sup.24, where R.sup.24 is selected
from the groups below:
##STR00482## [1167] and R.sup.25 and R.sup.26 are independently
selected from hydrogen or alkyl with the proviso that R.sup.25 and
R.sup.26 taken together with the N atom, to which they are
connected, may form a heterocycloalkyl, heterocycloalkyl, e.g.,
C3-C10 heterocycloalkyl wherein the heterocycle comprising 1 or
more heteroatoms selected from O, N, and S; and
[1168] R.sup.47 is selected from H or alkyl, e.g., C1-C6 alkyl.
[1169] In certain embodiments, the PTM of formula IX comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula IX is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1170] In certain additional embodiments, one or more of R.sup.24,
R.sup.25, R.sup.26, R.sup.33, R.sup.35, R.sup.37, R.sup.38,
R.sup.39, R.sup.40 or R.sup.47 groups of formula IX is coupled to a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1171] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula X:
##STR00483##
[1172] wherein R.sup.33, R.sup.35, R.sup.40 and B are as described
above;
[1173] R.sup.48 is selected from H or alkyl, e.g., C1-C6 alkyl;
and
[1174] R.sup.49 is selected from H, alkyl, e.g., C1-C6 alkyl, aryl,
e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10 heteroaryl comprising 1
or more heteroatoms selected from O, N, and S, cycloalkyl, e.g.,
C3-C10 cycloalkyl, heterocycloalkyl, e.g., C3-C10 heterocycloalkyl
wherein the heterocycle comprising 1 or more heteroatoms selected
from O, N, and S, arylalkyl, e.g., C5-C10 arylalkyl,
heteroarylalkyl, e.g., C5-C10 heteroarylalkyl wherein the
heteroaryl comprises 1 or more heteroatoms selected from O, N, and
S, cycloalkylalkyl or heterocycloalkylalkyl.
[1175] In a more preferred embodiment, PTMs of the PROTACs as
described herein comprise the moiety represented by the formula
X:
[1176] wherein R.sup.33 is selected from methoxy or ethoxy;
[1177] R.sup.35 is --NR.sup.37R.sup.38 where R.sup.37 and R.sup.38
are independently selected from H or alkyl, e.g., C1-C6 alkyl, or
--NR.sup.37R.sup.38 taken together represent a heterocycloalkyl
ring, most preferably piperazine, further optionally substituted
with alkyl, e.g., C1-C6 alkyl, amino, alkylamino, dialkylamino,
aminoalkyl, or --C(O)R.sup.39 where R.sup.39 is an alkyl, e.g.,
C1-C6 alkyl;
[1178] B is phenyl or a pyridyl, most preferably phenyl;
[1179] R.sup.40 is --NHC(O)R.sup.24, where R.sup.24 is selected
from the groups below:
##STR00484##
[1180] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl with the proviso that R.sup.25 and R.sup.26 taken
together with the N atom, to which they are connected, may form a
heterocycloalkyl;
[1181] R.sup.48 is selected from H or alkyl, e.g., C1-C6 alkyl;
and
[1182] R.sup.49 is selected from H or alkyl, e.g., C1-C6 alkyl.
[1183] In certain embodiments, the PTM of formula X comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula X is coupled to
a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1184] In certain additional embodiments, one or more of R.sup.24,
R.sup.25, R.sup.26, R.sup.33, R.sup.35, R.sup.37, R.sup.38,
R.sup.39, R.sup.40, R.sup.48 or R.sup.49 groups of formula X is
coupled to a linker, wherein the linker is a chemical moiety as
described herein coupling the PTM to the ULM group.
[1185] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula XI:
##STR00485##
[1186] wherein R.sup.50 is an alkyl, e.g., C1-C6 alkyl, alkylamino,
dialkylamino, cycloalkyl, e.g., C3-C10 cycloalkyl,
heterocycloalkyl, e.g., C3-C10 heterocycloalkyl wherein the
heterocycle comprising 1 or more heteroatoms selected from O, N,
and S, wherein the said cycloalkyl or heterocycloalkyl are further
optionally substituted with halogen, alkoxy, e.g., C1-C6 alkoxy,
amino, alkylamino, dialkylamino or cyano with the proviso that any
two heteroatoms are separated by at least two carbon atoms; and
[1187] R.sup.51 is a hydrogen and R.sup.52 is --NHC(O)R.sup.53
where R.sup.53 is an aryl or a heteroaryl optionally substituted
with 1-2 substituents independently selected from alkyl, e.g.,
C1-C6 alkyl, halogen, or haloalkyl, e.g., C1-C6 haloalkyl, alkoxy,
e.g., C1-C6 alkoxy, and cyano, or R.sup.51 and R.sup.52 taken
together constitute a 5-6-membered aryl or heteroaryl ring further
optionally substituted with 1-2 substituents independently selected
from alkyl, e.g., C1-C6 alkyl, halogen, or haloalkyl, e.g., C1-C6
haloalkyl, alkoxy, e.g., C1-C6 alkoxy, cyano, amino, alkylamino,
dilkylamino, aryl, e.g., C5-C10 aryl, or heteroaryl, e.g., C3-C10
heteroaryl comprising 1 or more heteroatoms selected from O, N, and
S.
[1188] In certain embodiments, the PTM of formula XI comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula XI is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1189] In certain additional embodiments, one or more of R.sup.50,
R.sup.51 or R.sup.52 groups of formula XI is coupled to a linker,
wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1190] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula XII:
##STR00486##
[1191] wherein G is a cycloalkyl, e.g., C3-C10 cycloalkyl, or
heterocycloalkyl, e.g., C3-C10 heterocycloalkyl wherein the
heterocycle comprising 1 or more heteroatoms selected from O, N,
and S;
[1192] H is an aryl, e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10
heteroaryl comprising 1 or more heteroatoms selected from O, N, and
S;
[1193] R.sup.43 is as described above; and
[1194] R.sup.53 and R.sup.54 are each independently 1-2
substituents independently selected from H, halogen, Cl, F, alkyl,
e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6 haloalkyl, alkenyl, e.g.,
C2-C6 alkenyl, alkynyl, e.g., C2-C6 alkynyl, alkoxy, e.g., C1-C6
alkoxy, amino, alkylamino, e.g., C1-C6 alkylamino, dialkylamino,
cyano, aryl, e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10 heteroaryl
comprising 1 or more heteroatoms selected from O, N, and S,
cycloalkyl, e.g., C3-C10 cycloalkyl, heterocycloalkyl, e.g., C3-C10
heterocycloalkyl wherein the heterocycle comprising 1 or more
heteroatoms selected from O, N, and S, wherein the said aryl or
heteroaryl can be further substituted with 1 to 2 substituents
selected from alkyl, e.g., C1-C6 alkyl, halogen, or haloalkyl,
e.g., C1-C6 haloalkyl, hydroxyl, alkoxy, e.g., C1-C6 alkoxy, amino,
alkylamino, dialkylamino and cyano.
[1195] In certain embodiments, the PTM of formula XII comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula XII is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1196] In certain embodiments, PTMs of the PROTACs as described
herein comprise the moiety represented by the formula XII:
[1197] wherein G is a heterocycloalkyl, heterocycloalkyl, e.g.,
C3-C10 heterocycloalkyl wherein the heterocycle comprising 1 or
more heteroatoms selected from O, N, and S most preferably 6- or
7-membered heterocycloalkyl, most preferably hexahydroazepine;
[1198] H is phenyl or a pyridyl;
[1199] R.sup.43 is 1 to 2 substituents where one substituent is
--NHC(O)R.sup.24, or --C(O)R.sup.24 if attached to a ring nitrogen
of D, and the other is optionally selected from H, alkyl, e.g.,
C1-C6 alkyl, halogen, or haloalkyl, e.g., C1-C6 haloalkyl, alkoxy,
e.g., C1-C6 alkoxy, wherein R.sup.24 is selected from the groups
below:
##STR00487##
[1200] R.sup.25 and R.sup.26 are independently selected from
hydrogen or alkyl, e.g., C1-C6 alkyl, with the proviso that
R.sup.25 and R.sup.26 taken together with the N atom, to which they
are connected, may form a heterocycloalkyl; and
[1201] R.sup.53 and R.sup.54 are each independently 1-2
substituents independently selected from H, halogen, alkyl, e.g.,
C1-C6 alkyl, and haloalkyl, e.g., C1-C6 haloalkyl.
[1202] In certain additional embodiments, one or more of R.sup.24,
R.sup.25, R.sup.26, R.sup.43, R.sup.53 or R.sup.84 groups of
formula XII is coupled to a linker, wherein the linker is a
chemical moiety as described herein coupling the PTM to the ULM
group.
[1203] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula XIII:
##STR00488##
[1204] wherein A is a saturated or unsaturated 4-8 atom carbocyclic
or heterocyclic ring comprising 1-7 heteroatoms, e.g., O, S or
N;
[1205] R.sup.k1 is selected from H, alkyl, e.g., C1-C6 alkyl,
alkenyl, e.g., C2-C6 alkenyl, alkynyl, e.g., C2-C6 alkynyl, aryl,
e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10 heteroaryl comprising 1
or more heteroatoms selected from O, N, and S, cycloalkyl, e.g.,
C3-C10 cycloalkyl, wherein the said alkyl, aryl or heteroaryl can
be further substituted with 1 to 2 substituents selected from
alkyl, e.g., C1-C6 alkyl, halogen, or haloalkyl, e.g., C1-C6
haloalkyl, alkoxy, e.g., C1-C6 alkoxy, hydroxyl, alkoxy, hydroxyl,
amino, alkylamino, dialkylamino and cyano;
[1206] R.sup.k2 is selected from H, alkyl, e.g., C1-C6 alkyl, aryl,
e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10 heteroaryl comprising 1
or more heteroatoms selected from O, N, and S, cycloalkyl, e.g.,
C3-C10 cycloalkyl, wherein the said alkyl, aryl or heteroaryl can
be further substituted with 1 to 2 substituents selected from
alkyl, e.g., C1-C6 alkyl, halogen, or haloalkyl, e.g., C1-C6
haloalkyl, alkoxy, e.g., C1-C6 alkoxy, hydroxyl, amino, alkylamino,
dialkylamino and cyano;
[1207] R.sup.k3 and R.sup.k4 are independently selected from H,
hydroxyl, alkyl, e.g., C1-C6 alkyl, alkoxy, e.g., C1-C6 alkoxy,
aryl, e.g., C5-C10 aryl, --SO.sup.2R.sup.k2, or halogen wherein the
said hydroxyl, alkyl, aryl or alkoxy can be further substituted
with 1 to 2 substituents selected from
[1208] alkyl, e.g., C1-C6 alkyl, halogen, or haloalkyl, e.g., C1-C6
haloalkyl, alkoxy, e.g., C1-C6 alkoxy, hydroxyl, amino, alkylamino,
dialkylamino and cyano;
[1209] X is N or CH or C with a double bond to the neighbor atom in
the ring.
[1210] In certain embodiments, the PTM of formula XIII comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula XIII is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1211] In certain additional embodiments, one or more of R.sup.k1,
R.sup.k2, R.sup.k3 or R.sup.k4 groups of formula XIII is coupled to
a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1212] In certain embodiments of formula XIII, A is substituted
with an alkyl, e.g., C1-C6 alkyl, alkoxy, e.g., C1-C6 alkoxy.
[1213] In certain embodiments of formula XIII, A is pyrazole,
imidazole, pyrrole, ozazole, or thiazole.
[1214] In certain embodiments of formula XIII, A is a pyrazole and
at least one of R.sup.k3 or R.sup.k4 is:
##STR00489##
[1215] In certain embodiments, an R group of formula XIII comprises
a linker, wherein the linker is a chemical moiety coupling the PTM
to the ULM group.
[1216] In certain embodiments, the PTM of formula XIII comprises
the structure:
##STR00490##
[1217] wherein, X, R.sup.k1, R.sup.k2, R.sup.k3, R.sup.k4, and
R.sup.k1 are as above; and
[1218] n=0, 1 or 2.
[1219] In certain additional embodiments, one or more of R.sup.k1,
R.sup.k2, R.sup.k3 or R.sup.k4 groups of formula XIII is coupled to
a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1220] In certain embodiments, the PTM of formula XIII comprises
the structure:
##STR00491##
[1221] wherein X, R.sup.k1, R.sup.k2, R.sup.k3, and R.sup.k4 are as
above.
[1222] In certain additional embodiments, one or more of R.sup.k1,
R.sup.k2, R.sup.k3 or R.sup.k4 groups of formula XIII is coupled to
a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1223] In certain embodiments, the PTM of formula XIII comprises
the structure:
##STR00492##
[1224] wherein R.sup.k18 is selected from H, alkyl, e.g., C1-C6
alkyl, haloalkyl, e.g., C1-C6 haloalkyl, alkenyl, e.g., C2-C6
alkenyl, alkynyl, e.g., C2-C6 alkynyl, aryl, e.g., C5-C10 aryl,
heteroaryl, e.g., C3-C10 heteroaryl comprising 1 or more
heteroatoms selected from O, N, and S, wherein the said alkyl, aryl
or heteroaryl can be further substituted with 1 to 2 substituents
selected from alkyl, e.g., C1-C6 alkyl, halogen, or haloalkyl,
e.g., C1-C6 haloalkyl, alkoxy, e.g., C1-C6 alkoxy, hydroxyl, amino,
alkylamino, dialkylamino and cyano;
[1225] R.sup.k19 is either H, alkyl (linear or branched) or halogen
substituted alkyl with 1-6 C atoms
[1226] R.sup.k20 is either H, alkyl (linear or branched) or halogen
substituted alkyl with 1-6 C atoms R.sup.k21 is either H, CN,
halogen or alkyl (linear or branched) or halogen substituted alkyl
with 1-3 C atoms; and
[1227] X.dbd.CH or N.
[1228] In certain additional embodiments, one or more of R.sup.k18,
R.sup.k19, R.sup.k20 or R.sup.k21 groups of formula XIII is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1229] In certain embodiments, the PTM of formula XIII comprises
the structure:
##STR00493##
[1230] wherein R.sup.k19 is either H, alkyl (linear or branched) or
halogen substituted alkyl with 1-6 C atoms;
[1231] R.sup.k20 is either H, alkyl (linear or branched) or halogen
substituted alkyl with 1-6 C atoms;
[1232] R.sup.k21 is either H, halogen or alkyl (linear or branched)
or halogen substituted alkyl with 1-3 C atoms; and
[1233] Linker (L) is a bond or a chemical linker coupling the PTM
to a ULM.
[1234] In certain additional embodiments, one or more of R.sup.k19,
R.sup.k20 or R.sup.k21 groups of formula XIII is coupled to a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1235] In certain embodiments, the PTM of formula XIII comprises
the structure:
##STR00494##
[1236] wherein R.sup.k20 and R.sup.k21 are as above;
[1237] R.sup.k22 is either H, alkyl (linear or branched) or halogen
substituted alkyl with 1-6 C atoms; and Linker (L) is a bond or
chemical linker coupling the PTM to a ULM.
[1238] In certain additional embodiments, one or more of R.sup.k20,
R.sup.k21 or R.sup.k22 groups of formula XIII is coupled to a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1239] In certain embodiments, the PTM of formula XIII comprises
the structure:
##STR00495##
[1240] wherein R.sup.k22 is either H, alkyl (linear or branched) or
halogen substituted alkyl with 1-6 C atoms;
[1241] R.sup.k23 is either H, alkyl (linear, branched or cyclic)
with up to 6 carbon atoms, optionally substituted with halogen;
[1242] R.sup.k24 is either H, halogen, cyano or alkyl (linear or
branched) or halogen substituted alkyl with 1-3 C atoms;
[1243] R.sup.k5 is either H, alkyl (linear or branched), alkoxy
(linear or branched) or hydroxyl; and
[1244] Linker (L) is a bond or chemical linker coupling the PTM to
a ULM.
[1245] In certain embodiments of formula XIII, the linker moiety is
conjugated to at least one of R.sup.k1, R.sup.k2, R.sup.k3, and
R.sup.k4 groups of formula XIII, wherein the linker is a chemical
moiety as described herein coupling the PTM to the ULM group.
[1246] In certain embodiments, the PTM of formula XIII comprises
the structure:
##STR00496##
[1247] wherein R.sup.k22 and R.sup.k23 are as defined above;
and
[1248] R is 1-3 substituents independently selected from H,
halogen, C1-C6 alkyl, cyano, or haloalkyl, wherein linker (L) is a
bond or chemical linker coupling PTM to a ULM.
[1249] In certain additional embodiments, one or more of R,
R.sup.k22 or R.sup.k33 groups of formula XIII is coupled to a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1250] In certain embodiments, the PTM of formula XIII comprises
the structure:
##STR00497##
In certain embodiments, the PTM includes a linker, wherein the
linker is a chemical moiety coupling the PTM to the ULM group.
[1251] In certain embodiments, PTMs of the PROTACs as described
herein comprise the moiety represented by the formula XIV:
##STR00498##
[1252] wherein R.sup.ka is H, alkyl, e.g., C1-C6 alkyl, alkoxy,
e.g., C1-C6 alkoxy;
[1253] R.sup.kb is aryl, e.g., C5-C10 aryl, or heteroaryl
comprising one or two 5-7 membered rings and 1-4 heteroatoms
selected from N, O, and S, wherein the aryl and heteroaryl are each
optionally substituted with one or more groups selected from alkyl,
e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6 haloalkyl, alkoxy, e.g.,
C1-C6 alkoxy, haloalkoxy, e.g., C1-C6 haloalkoxy, halogen,
NO.sub.2, OH, CN, C(O)Y.sup.1, C(O)OY.sup.1, C(O)NY.sup.1Y.sup.2,
NY.sup.1Y.sup.2, cycloalkyl, e.g., C3-C10 cycloalkyl, heterocycle
comprising 5-7 membered ring and 1-3 heteroatoms selected from N,
O, and S, C6-C10 aryl, and heteroaryl comprising one or two 5-7
membered rings and 1-4 heteroatoms selected from N, O, and S,
wherein the alkyl cycloalkyl, heterocyclyl, aryl or heteroaryl are
each optionally substituted with one or more Y.sup.1;
[1254] each Y.sup.1 and Y.sup.2 is independently selected from H,
alkyl, e.g., C1-C6 alkyl, haloalkyl, e.g., C1-C6 haloalkyl, alkoxy,
e.g., C1-C6 alkoxy, haloalkoxy, e.g., C1-C6 haloalkoxy, halogen,
NO.sub.2, OH, or CN;
[1255] wherein R.sup.k5, R.sup.k6 and R.sup.k7 are independently
selected from H, alkyl, e.g., C1-C6 alkyl, a alkyne, e.g., C1-C6
alkyne, alkoxy, e.g., C1-C6 alkoxy, hydroxyl, cyano, haloalkyl,
e.g., C1-C6 haloalkyl, haloalkoxy, e.g., C1-C6 haloalkoxy,
NO.sub.2, or halogen, Cl, F wherein the said alkyl or alkoxy can be
further substituted with 1 to 2 substituents selected from alkyl,
e.g., C1-C6 alkyl, halogen, or haloalkyl, e.g., C1-C6 haloalkyl,
alkoxy, e.g., C1-C6 alkoxy, hydroxyl, amino, alkylamino,
dialkylamino and cyano; and
[1256] X is N, CH, C(C1-C6 alkyl), C(C1-C6) haloalkyl, C(C1-6)
alkoxy, C(C1-C6) haloalkyl, halogen, C(NO.sub.2), C(NH.sub.2),
C(OH), or C(CN).
[1257] In certain embodiments, the PTM of formula XIV comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula XIV is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1258] In certain additional embodiments, one or more of R.sup.ka,
R.sup.kb, R.sup.k5, R.sup.k6 or R.sup.k7 groups of formula XIV is
coupled to a linker, wherein the linker is a chemical moiety as
described herein coupling the PTM to the ULM group.
[1259] In one embodiment of formula XIV, PTMs of the PROTACs as
described herein comprise the moiety represented by the
formula:
##STR00499##
[1260] wherein R.sup.k5, R.sup.k6 and R.sup.k7 are independently
selected from H, alkyl, e.g., C1-C6 alkyl, a alkyne, e.g., C1-C6
alkyne, alkoxy, e.g., C1-C6 alkoxy, hydroxyl, cyano, haloalkyl,
e.g., C1-C6 haloalkyl, haloalkoxy, e.g., C1-C6 haloalkoxy,
NO.sub.2, or halogen, Cl, F, wherein the said alkyl or alkoxy can
be further substituted with 1 to 2 substituents selected from
alkyl, e.g., C1-C6 alkyl, halogen, or haloalkyl, e.g., C1-C6
haloalkyl, alkoxy, e.g., C1-C6 alkoxy, hydroxyl, amino, alkylamino,
dialkylamino and cyano; and
[1261] X is N, CH, C(C1-C6 alkyl), C(C1-C6) haloalkyl, C(C1-6)
alkoxy, C(C1-C6) haloalkyl, halogen, C(NO.sub.2), C(NH.sub.2),
C(OH), or C(CN).
[1262] In certain additional embodiments, one or more of R.sup.k5,
R.sup.k6, or R.sup.k7 groups of formula XIV is coupled to a linker,
wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1263] In certain embodiments, the PTM of formula XIV is linked to
a ULM via an unsaturated chemical linker. In certain embodiments,
the unsaturated linker is substituted.
[1264] In certain embodiments, the PTM of formula XIV comprises the
structure:
##STR00500##
[1265] wherein R.sup.k26 and R.sup.k27 are independently selected
from H, alkyl, e.g., C1-C6 alkyl, alkoxy, e.g., C1-C6 alkoxy,
hydroxyl, cyano or halogen, wherein the said alkyl or alkoxy can be
further substituted with 1 to 2 substituents selected from alkyl,
e.g., C1-C6 alkyl, halogen, or haloalkyl, e.g., C1-C6 haloalkyl,
alkoxy, e.g., C1-C6 alkoxy, hydroxyl, amino, alkylamino,
dialkylamino and cyano.
[1266] In certain additional embodiments, one or more of R.sup.k26
or R.sup.k27 groups of formula XIV is coupled to a linker, wherein
the linker is a chemical moiety as described herein coupling the
PTM to the ULM group.
[1267] In certain embodiments, the PTM of formula XIV comprises the
structure:
##STR00501##
[1268] wherein R.sup.k27 is selected from H, cyano or halogen,
wherein Linker (L) is a bond or a chemical linker coupling the PTM
to a ULM.
[1269] In certain embodiments, the PTM of formula XIV comprises the
structure:
##STR00502##
[1270] wherein R.sup.k27 is selected from H, cyano or halogen,
wherein Linker (L) is a bond or a chemical linker coupling the PTM
to a ULM.
[1271] In certain additional embodiments, R.sup.k27 of formula XIV
is coupled to a linker, wherein the linker is a chemical moiety as
described herein coupling the PTM to the ULM group.
[1272] In certain embodiments, the PTM of formula XIV comprises the
structure:
##STR00503##
[1273] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula XV:
##STR00504##
[1274] wherein R.sup.k8 is selected from H, alkyl, e.g., C1-C6
alkyl, haloalkyl, e.g., C1-C6 haloalkyl, alkenyl, e.g., C2-C6
alkenyl, alkynyl, e.g., C2-C6 alkynyl, alkoxy, e.g., C1-C6 alkoxy,
aryl, e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10 heteroaryl
comprising 1 or more heteroatoms selected from O, N, and S, wherein
the said alkyl, aryl or heteroaryl can be further substituted with
1 to 2 substituents selected from alkyl, e.g., C1-C6 alkyl,
halogen, haloalkyl, e.g., C1-C6 haloalkyl, hydroxyl, alkoxy, e.g.,
C1-C6 alkoxy, amino, alkylamino, dialkylamino and cyano;
[1275] R.sup.k9 is selected from H, alkyl, e.g., C1-C6 alkyl or
cycloalkyl, e.g., C3-C10 cycloalkyl, wherein the said alkyl or
cycloalkyl can be further substituted with 1 to 3 substituents
selected from alkyl, e.g., C1-C6 alkyl, halogen, or haloalkyl,
e.g., C1-C6 haloalkyl, alkoxy, e.g., C1-C6 alkoxy, hydroxyl, amino,
alkylamino, dialkylamino and cyano; and
[1276] R.sup.k10 is selected from H, alkyl, e.g., C1-C6 alkyl,
alkylsulfone, alkylcarboxamide or aryl, wherein the said alkyl or
aryl can be further substituted with 1 to 2 substituents selected
from alkyl, e.g., C1-C6 alkyl, halogen, alkylsulfone,
alkylsulfonamide, amide, carboxamide, haloalkyl, e.g., C1-C6
haloalkyl, hydroxyl, alkoxy, e.g., C1-C6 alkoxy, amino, amide,
alkylamino, dialkylamino and cyano.
[1277] In certain embodiments, the PTM of formula XV comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula XV is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1278] In certain additional embodiments, one or more of R.sup.k8,
R.sup.k9, or R.sup.k10 groups of formula XV is coupled to a linker,
wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1279] In certain embodiments of formula XV, R.sup.k10 is:
##STR00505##
[1280] In certain embodiments, the PTM of formula XV comprises the
structure:
##STR00506##
[1281] wherein R.sup.k28 is selected from H, alkyl, e.g., C1-C6
alkyl, haloalkyl, e.g., C1-C6 haloalkyl, alkenyl, e.g., C2-C6
alkenyl, alkynyl, e.g., C2-C6 alkynyl, alkoxy, e.g., C1-C6 alkoxy,
aryl, e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10 heteroaryl
comprising 1 or more heteroatoms selected from O, N, and S, wherein
the said alkyl, aryl or heteroaryl can be further substituted with
1 to 2 substituents selected from alkyl, e.g., C1-C6 alkyl,
halogen, or haloalkyl, e.g., C1-C6 haloalkyl, alkoxy, e.g., C1-C6
alkoxy, hydroxyl, amino, alkylamino, dialkylamino and cyano;
[1282] R.sup.k29 is selected from H, alkyl, e.g., C1-C6 alkyl, or
cycloalkyl, e.g., C3-C10 cycloalkyl, wherein the said alkyl or
cycloalkyl can be further substituted with 1-3 halogen
substituents; and
[1283] R.sup.k30 is selected from H, alkyl, e.g., C1-C6 alkyl,
cycloalkyl, e.g., C3-C10 cycloalkyl, alkylsulfone,
cycloalkylsulfone or --COR--wherein the said alkyl or aryl can be
further substituted with 1 to 2 substituents selected from alkyl,
e.g., C1-C6 alkyl, halogen, or haloalkyl, e.g., C1-C6 haloalkyl,
alkoxy, e.g., C1-C6 alkoxy, hydroxyl, amino, amide, alkylamino,
dialkylamino and cyano.
[1284] In certain additional embodiments, one or more of R.sup.k28,
R.sup.k29, or R.sup.k30 groups of formula XV is coupled to a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1285] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula XVI:
##STR00507##
[1286] wherein R.sup.k11 is selected from H, alkyl, e.g., C1-C6
alkyl, alkoxy, e.g., C1-C6 alkoxy, C(O)NHR where R is selected from
H, alkyl, e.g., C1-C6 alkyl, cycloalkyl, e.g., C3-C10 cycloalkyl or
a saturated heterocycle with 4-6 ring atoms;
[1287] R.sup.k12 is selected from H, alkyl (linear or branched),
e.g., C1-C6 alkyl, aryl, e.g., C5-C10 aryl, heteroaryl, e.g.,
C3-C10 heteroaryl comprising 1 or more heteroatoms selected from O,
N, and S, cycloalkyl, e.g., C3-C10 cycloalkyl, wherein the said
alkyl, aryl or heteroaryl can be further substituted with 1 to 2
substituents selected from alkyl, e.g., C1-C6 alkyl, halogen, or
haloalkyl, e.g., C1-C6 haloalkyl, alkoxy, e.g., C1-C6 alkoxy,
hydroxyl, amino, alkylamino, dialkylamino and cyano;
[1288] R.sup.k13 is selected from H, alkyl, --C(O)NHR, C(O)R,
S(O).sub.2R where R is H, alkyl, e.g., C1-C6 alkyl, or cycloalkyl,
e.g., C3-C10 cycloalkyl, which can be further substituted with 1 to
2 substituents selected from alkyl, e.g., C1-C6 alkyl, halogen,
alkylsulfone, alkylsulfonamide, amide, carboxamide, haloalkyl,
e.g., C1-C6 haloalkyl, alkoxy, e.g., C1-C6 alkoxy, hydroxyl, amino,
amide, alkylamino, dialkylamino and cyano; and
[1289] X is N or CH.
[1290] In certain embodiments, the PTM of formula XVI comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula XVI is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1291] In certain additional embodiments, one or more of R.sup.k11,
R.sup.k12, or R.sup.k13 groups of formula XVI is coupled to a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1292] In certain embodiments of formula XVI, R.sup.k13 is:
##STR00508##
[1293] In certain embodiments, an R group of formula XVI comprises
a linker, wherein the linker is a chemical moiety coupling the PTM
to the ULM group.
[1294] In certain embodiments, the PTM of formula XVI comprises the
structure:
##STR00509##
[1295] wherein R.sup.k31 is selected from H, alkyl, e.g., C1-C6
alkyl, alkoxy, C(O)NHR where R is selected from H, alkyl, e.g.,
C1-C6 alkyl, cycloalkyl, e.g., C3-C10 cycloalkyl or a saturated
heterocycle with 4-6 ring atoms;
[1296] R.sup.k32 is selected from H, alkyl (linear or branched),
e.g., C1-C6 alkyl, or halogen substituted alkyl with 1-6 C
atoms;
[1297] R.sup.k33 is selected from H, alkyl, --C(O)NHR, C(O)R,
S(O).sub.2R where R is H, alkyl, e.g., C1-C6 alkyl, or cycloalkyl,
which can be further substituted with 1 to 2 substituents selected
from alkyl, e.g., C1-C6 alkyl, halogen, alkylsulfone,
alkylsulfonamide, amide, carboxamide, haloalkyl, e.g., C1-C6
haloalkyl, hydroxyl, alkoxy, e.g., C1-C6 alkoxy, amino, amide,
alkylamino, dialkylamino and cyano; and
[1298] X is N or CH.
[1299] In certain additional embodiments, one or more of R.sup.k31,
R.sup.k32, or R.sup.k33 groups of formula XVI is coupled to a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group.
[1300] In certain embodiments of formula XVI, R.sup.k33 is:
##STR00510##
[1301] In one embodiment, PTMs of the PROTACs as described herein
comprise the moiety represented by the formula XVII:
##STR00511##
[1302] wherein R.sup.k14 is selected from H, alkyl, e.g., C1-C6
alkyl, alkoxy, e.g., C1-C6 alkoxy, --C(O)NHR where R is selected
from H, alkyl, e.g., C1-C6 alkyl, cycloalkyl, e.g., C3-C10
cycloalkyl, or a saturated heterocycle with 4-6 ring atoms;
[1303] R.sup.k15 is selected from H, alkyl (linear or branched),
aryl, e.g., C5-C10 aryl, heteroaryl, e.g., C3-C10 heteroaryl
comprising 1 or more heteroatoms selected from O, N, and S,
cycloalkyl, e.g., C3-C10 cycloalkyl, wherein the said alkyl, aryl
or heteroaryl can be further substituted with 1 to 2 substituents
selected from alkyl, e.g., C1-C6 alkyl, halogen, or haloalkyl,
e.g., C1-C6 haloalkyl, alkoxy, e.g., C1-C6 alkoxy, hydroxyl, amino,
alkylamino, dialkylamino and cyano;
[1304] R.sup.k16 is selected from H, alkyl, e.g., C1-C6 alkyl, or
cycloalkyl, e.g., C3-C10 cycloalkyl, which can be further
substituted with 1 to 2 substituents selected from alkyl, e.g.,
C1-C6 alkyl, halogen, alkylsulfone, alkylsulfonamide, amide,
carboxamide, haloalkyl, e.g., C1-C6 haloalkyl, alkoxy, e.g., C1-C6
alkoxy, hydroxyl, amino, amide, alkylamino, dialkylamino and cyano;
and
[1305] R.sup.k17 is selected from H, halogen, CN.
[1306] In certain embodiments, the PTM of formula XVII comprises a
linker, wherein the linker is a chemical moiety as described herein
coupling the PTM to the ULM group. In certain additional
embodiments, one or more of the R groups of formula XVII is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1307] In certain additional embodiments, one or more of R.sup.k14,
R.sup.k15, R.sup.k16 or R.sup.k17 groups of formula XVII is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1308] In certain embodiments, the PTM of formula XVII comprises
the structure:
##STR00512##
[1309] wherein R.sup.k31 is selected from H, alkyl, alkoxy, C(O)NHR
where R is selected from H, alkyl, cycloalkyl or a saturated
heterocycle with 4-6 ring atoms;
[1310] R.sup.k32 is selected from H, alkyl (linear or branched) or
halogen substituted alkyl with 1-6 C atoms;
[1311] R.sup.k34 is selected from H, alkyl (linear or branched) or
halogen substituted alkyl with 1-3 C atoms; and
[1312] R.sup.k35 is selected from H, F, Cl, Br or cyano.
[1313] In certain additional embodiments, one or more of R.sup.k31,
R.sup.k32, R.sup.k34 or R.sup.k35 groups of formula XVII is coupled
to a linker, wherein the linker is a chemical moiety as described
herein coupling the PTM to the ULM group.
[1314] In certain embodiments, the PTM of formula XVII comprises
the structure:
##STR00513##
[1315] As described herein, the PTMs are coupled via a linker
moiety to a ULM. It is contemplated that the linker moiety can be
conjugated at any location desired on the PTM. In certain
embodiments, the PTM comprises a structure selected from the group
consisting of:
##STR00514## ##STR00515## ##STR00516## ##STR00517##
[1316] In certain embodiments, the description provides a compound
having the structure selected from compound 1-351 as described in
FIG. 2.
Therapeutic Compositions
[1317] Pharmaceutical compositions comprising combinations of an
effective amount of at least one bifunctional compound as described
herein, and one or more of the compounds otherwise described
herein, all in effective amounts, in combination with a
pharmaceutically effective amount of a carrier, additive or
excipient, represents a further aspect of the present
disclosure.
[1318] In certain embodiments, the description provides a
therapeutic composition comprising an effective amount of at least
one compound selected from compound 1-351 as described in FIG. 2,
and a pharmaceutically acceptable carrier or excipient. In certain
embodiments, the description provides a combination for
co-administration (e.g., either separately or in a single dosage
form) comprising an effective amount of at least one compound as
described herein, at least one additional bioactive agent, and a
pharmaceutically acceptable carrier or excipient. In certain
embodiments, the additional bioactive agent is an anti-oncologic
agent.
[1319] The present disclosure includes, where applicable, the
compositions comprising the pharmaceutically acceptable salts, in
particular, acid or base addition salts of compounds as described
herein. The acids which are used to prepare the pharmaceutically
acceptable acid addition salts of the aforementioned base compounds
useful according to this aspect are those which form non-toxic acid
addition salts, i.e., salts containing pharmacologically acceptable
anions, such as the hydrochloride, hydrobromide, hydroiodide,
nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate,
lactate, citrate, acid citrate, tartrate, bitartrate, succinate,
maleate, fumarate, gluconate, saccharate, benzoate,
methanesulfonate, ethanesulfonate, benzenesulfonate,
p-toluenesulfonate and pamoate [i.e.,
1,1'-methylene-bis-(2-hydroxy-3 naphthoate)]salts, among numerous
others.
[1320] Pharmaceutically acceptable base addition salts may also be
used to produce pharmaceutically acceptable salt forms of the
compounds or derivatives according to the present disclosure. The
chemical bases that may be used as reagents to prepare
pharmaceutically acceptable base salts of the present compounds
that are acidic in nature are those that form non-toxic base salts
with such compounds. Such non-toxic base salts include, but are not
limited to those derived from such pharmacologically acceptable
cations such as alkali metal cations (eg., potassium and sodium)
and alkaline earth metal cations (eg, calcium, zinc and magnesium),
ammonium or water-soluble amine addition salts such as
N-methylglucamine-(meglumine), and the lower alkanolammonium and
other base salts of pharmaceutically acceptable organic amines,
among others.
[1321] The compounds as described herein may, in accordance with
the disclosure, be administered in single or divided doses by the
oral, parenteral or topical routes. Administration of the active
compound may range from continuous (intravenous drip) to several
oral administrations per day (for example, Q.I.D.) and may include
oral, topical, parenteral, intramuscular, intravenous,
sub-cutaneous, transdermal (which may include a penetration
enhancement agent), buccal, sublingual and suppository
administration, among other routes of administration. Enteric
coated oral tablets may also be used to enhance bioavailability of
the compounds from an oral route of administration. The most
effective dosage form will depend upon the pharmacokinetics of the
particular agent chosen as well as the severity of disease in the
patient. Administration of compounds according to the present
disclosure as sprays, mists, or aerosols for intra-nasal,
intra-tracheal or pulmonary administration may also be used. The
present disclosure therefore also is directed to pharmaceutical
compositions comprising an effective amount of compound as
described herein, optionally in combination with a pharmaceutically
acceptable carrier, additive or excipient. Compounds according to
the present disclosure may be administered in immediate release,
intermediate release or sustained or controlled release forms.
Sustained or controlled release forms are preferably administered
orally, but also in suppository and transdermal or other topical
forms. Intramuscular injections in liposomal form may also be used
to control or sustain the release of compound at an injection
site.
[1322] In another aspect, the disclosure provides therapeutic
compositions comprising an effective and/or synergistic amount of a
compound as described herein, and at least one additional bioactive
agent (i.e., a combination therapeutic), e.g., another EGFR PROTAC
as described herein, an anticancer, and anti-inflammatory, and/or
an EGFR inhibitor. In certain embodiments, the EGFR inhibitor
includes at least one of gefitinib, erlotinib, afatinib,
brigatinib, icotinib, lapatinib, cetuximab, panitumumab,
osimertinib, zalutumumab, nimotuzumab, matuzumab or combinations
thereof. In certain embodiments, the combination therapy
composition an effective amount of the EGFR inhibitor.
[1323] The compositions as described herein may be formulated in a
conventional manner using one or more pharmaceutically acceptable
carriers and may also be administered in controlled-release
formulations. Pharmaceutically acceptable carriers that may be used
in these pharmaceutical compositions include, but are not limited
to, ion exchangers, alumina, aluminum stearate, lecithin, serum
proteins, such as human serum albumin, buffer substances such as
phosphates, glycine, sorbic acid, potassium sorbate, partial
glyceride mixtures of saturated vegetable fatty acids, water, salts
or electrolytes, such as prolamine sulfate, disodium hydrogen
phosphate, potassium hydrogen phosphate, sodium chloride, zinc
salts, colloidal silica, magnesium trisilicate, polyvinyl
pyrrolidone, cellulose-based substances, polyethylene glycol,
sodium carboxymethylcellulose, polyacrylates, waxes,
polyethylene-polyoxypropylene-block polymers, polyethylene glycol
and wool fat.
[1324] The compositions as described herein may be administered
orally, parenterally, by inhalation spray, topically, rectally,
nasally, buccally, vaginally or via an implanted reservoir. The
term "parenteral" as used herein includes subcutaneous,
intravenous, intramuscular, intra-articular, intra-synovial,
intrasternal, intrathecal, intrahepatic, intralesional and
intracranial injection or infusion techniques. Preferably, the
compositions are administered orally, intraperitoneally or
intravenously.
[1325] Sterile injectable forms of the compositions as described
herein may be aqueous or oleaginous suspension. These suspensions
may be formulated according to techniques known in the art using
suitable dispersing or wetting agents and suspending agents. The
sterile injectable preparation may also be a sterile injectable
solution or suspension in a non-toxic parenterally-acceptable
diluent or solvent, for example as a solution in 1, 3-butanediol.
Among the acceptable vehicles and solvents that may be employed are
water, Ringer's solution and isotonic sodium chloride solution. In
addition, sterile, fixed oils are conventionally employed as a
solvent or suspending medium. For this purpose, any bland fixed oil
may be employed including synthetic mono- or di-glycerides. Fatty
acids, such as oleic acid and its glyceride derivatives are useful
in the preparation of injectables, as are natural
pharmaceutically-acceptable oils, such as olive oil or castor oil,
especially in their polyoxyethylated versions. These oil solutions
or suspensions may also contain a long-chain alcohol diluent or
dispersant, such as Ph. Helv or similar alcohol.
[1326] The pharmaceutical compositions as described herein may be
orally administered in any orally acceptable dosage form including,
but not limited to, capsules, tablets, aqueous suspensions or
solutions. In the case of tablets for oral use, carriers which are
commonly used include lactose and corn starch. Lubricating agents,
such as magnesium stearate, are also typically added. For oral
administration in a capsule form, useful diluents include lactose
and dried corn starch. When aqueous suspensions are required for
oral use, the active ingredient is combined with emulsifying and
suspending agents. If desired, certain sweetening, flavoring or
coloring agents may also be added.
[1327] Alternatively, the pharmaceutical compositions as described
herein may be administered in the form of suppositories for rectal
administration. These can be prepared by mixing the agent with a
suitable non-irritating excipient, which is solid at room
temperature but liquid at rectal temperature and therefore will
melt in the rectum to release the drug. Such materials include
cocoa butter, beeswax and polyethylene glycols.
[1328] The pharmaceutical compositions as described herein may also
be administered topically. Suitable topical formulations are
readily prepared for each of these areas or organs. Topical
application for the lower intestinal tract can be effected in a
rectal suppository formulation (see above) or in a suitable enema
formulation. Topically-acceptable transdermal patches may also be
used.
[1329] For topical applications, the pharmaceutical compositions
may be formulated in a suitable ointment containing the active
component suspended or dissolved in one or more carriers. Carriers
for topical administration of the compounds of this disclosure
include, but are not limited to, mineral oil, liquid petrolatum,
white petrolatum, propylene glycol, polyoxyethylene,
polyoxypropylene compound, emulsifying wax and water. In certain
preferred aspects of the disclosure, the compounds may be coated
onto a stent which is to be surgically implanted into a patient in
order to inhibit or reduce the likelihood of occlusion occurring in
the stent in the patient.
[1330] Alternatively, the pharmaceutical compositions can be
formulated in a suitable lotion or cream containing the active
components suspended or dissolved in one or more pharmaceutically
acceptable carriers. Suitable carriers include, but are not limited
to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl
esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and
water.
[1331] For ophthalmic use, the pharmaceutical compositions may be
formulated as micronized suspensions in isotonic, pH adjusted
sterile saline, or, preferably, as solutions in isotonic, pH
adjusted sterile saline, either with our without a preservative
such as benzylalkonium chloride. Alternatively, for ophthalmic
uses, the pharmaceutical compositions may be formulated in an
ointment such as petrolatum.
[1332] The pharmaceutical compositions as described herein may also
be administered by nasal aerosol or inhalation. Such compositions
are prepared according to techniques well-known in the art of
pharmaceutical formulation and may be prepared as solutions in
saline, employing benzyl alcohol or other suitable preservatives,
absorption promoters to enhance bioavailability, fluorocarbons,
and/or other conventional solubilizing or dispersing agents.
[1333] The amount of compound in a pharmaceutical composition as
described herein that may be combined with the carrier materials to
produce a single dosage form will vary depending upon the host and
disease treated, the particular mode of administration. Preferably,
the compositions should be formulated to contain between about 0.05
milligram to about 750 milligrams or more, more preferably about 1
milligram to about 600 milligrams, and even more preferably about
10 milligrams to about 500 milligrams of active ingredient, alone
or in combination with at least one other compound according to the
present disclosure.
[1334] It should also be understood that a specific dosage and
treatment regimen for any particular patient will depend upon a
variety of factors, including the activity of the specific compound
employed, the age, body weight, general health, sex, diet, time of
administration, rate of excretion, drug combination, and the
judgment of the treating physician and the severity of the
particular disease or condition being treated.
[1335] A patient or subject in need of therapy using compounds
according to the methods described herein can be treated by
administering to the patient (subject) an effective amount of the
compound according to the present disclosure including
pharmaceutically acceptable salts, solvates or polymorphs, thereof
optionally in a pharmaceutically acceptable carrier or diluent,
either alone, or in combination with other known erythopoiesis
stimulating agents as otherwise identified herein.
[1336] These compounds can be administered by any appropriate
route, for example, orally, parenterally, intravenously,
intradermally, subcutaneously, or topically, including
transdermally, in liquid, cream, gel, or solid form, or by aerosol
form.
[1337] The active compound is included in the pharmaceutically
acceptable carrier or diluent in an amount sufficient to deliver to
a patient a therapeutically effective amount for the desired
indication, without causing serious toxic effects in the patient
treated. A preferred dose of the active compound for all of the
herein-mentioned conditions is in the range from about 10 ng/kg to
300 mg/kg, preferably 0.1 to 100 mg/kg per day, more generally 0.5
to about 25 mg per kilogram body weight of the recipient/patient
per day. A typical topical dosage will range from 0.01-5% wt/wt in
a suitable carrier.
[1338] The compound is conveniently administered in any suitable
unit dosage form, including but not limited to one containing less
than 1 mg, 1 mg to 3000 mg, preferably 5 to 500 mg of active
ingredient per unit dosage form. An oral dosage of about 25-250 mg
is often convenient.
[1339] The active ingredient is preferably administered to achieve
peak plasma concentrations of the active compound of about
0.00001-30 mM, preferably about 0.1-30 .mu.M. This may be achieved,
for example, by the intravenous injection of a solution or
formulation of the active ingredient, optionally in saline, or an
aqueous medium or administered as a bolus of the active ingredient.
Oral administration is also appropriate to generate effective
plasma concentrations of active agent.
[1340] The concentration of active compound in the drug composition
will depend on absorption, distribution, inactivation, and
excretion rates of the drug as well as other factors known to those
of skill in the art. It is to be noted that dosage values will also
vary with the severity of the condition to be alleviated. It is to
be further understood that for any particular subject, specific
dosage regimens should be adjusted over time according to the
individual need and the professional judgment of the person
administering or supervising the administration of the
compositions, and that the concentration ranges set forth herein
are exemplary only and are not intended to limit the scope or
practice of the claimed composition. The active ingredient may be
administered at once, or may be divided into a number of smaller
doses to be administered at varying intervals of time.
[1341] Oral compositions will generally include an inert diluent or
an edible carrier. They may be enclosed in gelatin capsules or
compressed into tablets. For the purpose of oral therapeutic
administration, the active compound or its prodrug derivative can
be incorporated with excipients and used in the form of tablets,
troches, or capsules. Pharmaceutically compatible binding agents,
and/or adjuvant materials can be included as part of the
composition.
[1342] The tablets, pills, capsules, troches and the like can
contain any of the following ingredients, or compounds of a similar
nature: a binder such as microcrystalline cellulose, gum tragacanth
or gelatin; an excipient such as starch or lactose, a dispersing
agent such as alginic acid, Primogel, or corn starch; a lubricant
such as magnesium stearate or Sterotes; a glidant such as colloidal
silicon dioxide; a sweetening agent such as sucrose or saccharin;
or a flavoring agent such as peppermint, methyl salicylate, or
orange flavoring. When the dosage unit form is a capsule, it can
contain, in addition to material of the above type, a liquid
carrier such as a fatty oil. In addition, dosage unit forms can
contain various other materials which modify the physical form of
the dosage unit, for example, coatings of sugar, shellac, or
enteric agents.
[1343] The active compound or pharmaceutically acceptable salt
thereof can be administered as a component of an elixir,
suspension, syrup, wafer, chewing gum or the like. A syrup may
contain, in addition to the active compounds, sucrose as a
sweetening agent and certain preservatives, dyes and colorings and
flavors.
[1344] The active compound or pharmaceutically acceptable salts
thereof can also be mixed with other active materials that do not
impair the desired action, or with materials that supplement the
desired action, such as erythropoietin stimulating agents,
including EPO and darbapoietin alfa, among others. In certain
preferred aspects of the disclosure, one or more compounds
according to the present disclosure are coadministered with another
bioactive agent, such as an erythropoietin stimulating agent or a
would healing agent, including an antibiotic, as otherwise
described herein.
[1345] Solutions or suspensions used for parenteral, intradermal,
subcutaneous, or topical application can include the following
components: a sterile diluent such as water for injection, saline
solution, fixed oils, polyethylene glycols, glycerine, propylene
glycol or other synthetic solvents; antibacterial agents such as
benzyl alcohol or methyl parabens; antioxidants such as ascorbic
acid or sodium bisulfite; chelating agents such as
ethylenediaminetetraacetic acid; buffers such as acetates, citrates
or phosphates and agents for the adjustment of tonicity such as
sodium chloride or dextrose. The parental preparation can be
enclosed in ampoules, disposable syringes or multiple dose vials
made of glass or plastic.
[1346] If administered intravenously, preferred carriers are
physiological saline or phosphate buffered saline (PBS).
[1347] In one embodiment, the active compounds are prepared with
carriers that will protect the compound against rapid elimination
from the body, such as a controlled release formulation, including
implants and microencapsulated delivery systems. Biodegradable,
biocompatible polymers can be used, such as ethylene vinyl acetate,
polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and
polylactic acid. Methods for preparation of such formulations will
be apparent to those skilled in the art.
[1348] Liposomal suspensions may also be pharmaceutically
acceptable carriers. These may be prepared according to methods
known to those skilled in the art, for example, as described in
U.S. Pat. No. 4,522,811 (which is incorporated herein by reference
in its entirety). For example, liposome formulations may be
prepared by dissolving appropriate lipid(s) (such as stearoyl
phosphatidyl ethanolamine, stearoyl phosphatidyl choline,
arachadoyl phosphatidyl choline, and cholesterol) in an inorganic
solvent that is then evaporated, leaving behind a thin film of
dried lipid on the surface of the container. An aqueous solution of
the active compound is then introduced into the container. The
container is then swirled by hand to free lipid material from the
sides of the container and to disperse lipid aggregates, thereby
forming the liposomal suspension.
Therapeutic Methods
[1349] In an additional aspect, the description provides
therapeutic compositions comprising an effective amount of a
compound as described herein or salt form thereof, and a
pharmaceutically acceptable carrier. The therapeutic compositions
modulate protein degradation in a patient or subject, for example,
an animal such as a human, and can be used for treating or
ameliorating disease states or conditions which are modulated
through the degraded protein.
[1350] The terms "treat", "treating", and "treatment", etc., as
used herein, refer to any action providing a benefit to a patient
for which the present compounds may be administered, including the
treatment of any disease state or condition which is modulated
through the protein to which the present compounds bind. Disease
states or conditions, e.g., EGFR-related diseases or disorders,
which may be treated using compounds according to the present
disclosure are contemplated.
[1351] The description provides therapeutic compositions as
described herein for effectuating the degradation of proteins of
interest for the treatment or amelioration of an EGFR-related
disease, e.g., cancer and/or an inflammatory disorder, and/or hair
growth. As such, in another aspect, the description provides a
method of ubiquitinating/degrading a target protein in a cell.
[1352] In certain embodiments, the description provides
compositions and methods for treating an EGFR-related disease or
disorder. In certain embodiments, the EGFR-related disease or
disorder is at least one of cancer and/or an inflammatory disorder.
In certain embodiments, the EGFR-related disease or disorder is at
least one of squamous-cell carcinoma of the lung, colon and anal
cancers, glioblastoma, and epithelial tumors of the head and neck,
psoriasis, eczema and atherosclerosis or a combination thereof.
[1353] In certain embodiments, the method comprises administering a
bifunctional compound as described herein comprising, e.g., a ULM
and a PTM, preferably linked through a linker moiety, as otherwise
described herein, wherein the ULM is coupled to the PTM and wherein
the ULM recognizes a ubiquitin pathway protein (e.g., an ubiquitin
ligase, such as an E3 ubiquitin ligase including cereblon, VHL,
IAP, and/or MDM2) and the PTM recognizes the target protein such
that degradation of the target protein will occur when the target
protein is placed in proximity to the ubiquitin ligase, thus
resulting in degradation/inhibition of the effects of the target
protein and the control of protein levels. The control of protein
levels afforded by the present disclosure provides treatment of a
disease state or condition, which is modulated through the target
protein by lowering the level of that protein in the cell, e.g.,
cell of a patient. In certain embodiments, the method comprises
administering an effective amount of a compound as described
herein, optionally including a pharmaceutically acceptable
excipient, carrier, adjuvant, another bioactive agent or
combination thereof.
[1354] In additional embodiments, the description provides methods
for treating or ameliorating a disease, disorder or symptom thereof
in a subject or a patient, e.g., an animal such as a human,
comprising administering to a subject in need thereof a composition
comprising an effective amount, e.g., a therapeutically effective
amount, of a compound as described herein or salt form thereof, and
a pharmaceutically acceptable excipient, carrier, adjuvant, another
bioactive agent or combination thereof, wherein the composition is
effective for treating or ameliorating the disease or disorder or
symptom thereof in the subject.
[1355] In another aspect, the description provides methods for
identifying the effects of the degradation of proteins of interest
in a biological system using compounds according to the present
disclosure.
[1356] In another embodiment, the present disclosure is directed to
a method of treating a human patient in need for a disease state or
condition modulated through EGFR protein where the degradation of
that protein will produce a therapeutic effect in the patient, the
method comprising administering to a patient in need an effective
amount of a compound according to the present disclosure. In
certain embodiments, the compound as described herein is
administered in combination with another bioactive agent, e.g., an
EGFR inhibitor, as described herein.
[1357] The term "disease state or condition" is used to describe
any disease state or condition wherein EGFR protein dysregulation
(i.e., the amount of EGFR protein expressed in a patient is
elevated) occurs and where degradation of EGFR proteins in a
patient may provide beneficial therapy or relief of symptoms to a
patient in need thereof. In certain instances, the disease state or
condition may be cured.
[1358] The term "bioactive agent" is used to describe an agent,
other than a compound according to the present disclosure, which is
used in combination with the present compounds as an agent with
biological activity to assist in effecting an intended therapy,
inhibition and/or prevention/prophylaxis for which the present
compounds are used. Preferred bioactive agents for use herein
include those agents which have pharmacological activity similar to
that for which the present compounds are used or administered and
include for example, anti-cancer agents, antiviral agents,
especially including anti-HIV agents and anti-HCV agents,
antimicrobial agents, antifungal agents, etc.
[1359] The term "additional anti-cancer agent" is used to describe
an anti-cancer agent, which may be combined with compounds
according to the present disclosure to treat cancer. These agents
include, for example, everolimus, trabectedin, abraxane, TLK 286,
AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON 0910.Na, AZD 6244
(ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin,
vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263,
a FLT-3 inhibitor, a VEGFR inhibitor, an EGFR TK inhibitor, an
aurora kinase inhibitor, a PIK-1 modulator, a Bcl-2 inhibitor, an
HDAC inhbitor, a c-MET inhibitor, a PARP inhibitor, a Cdk
inhibitor, an EGFR TK inhibitor, an IGFR-TK inhibitor, an anti-HGF
antibody, a PI3 kinase inhibitor, an AKT inhibitor, an mTORC1/2
inhibitor, a JAK/STAT inhibitor, a checkpoint-1 or 2 inhibitor, a
focal adhesion kinase inhibitor, a Map kinase kinase (mek)
inhibitor, a VEGF trap antibody, pemetrexed, erlotinib, dasatanib,
nilotinib, decatanib, panitumumab, amrubicin, oregovomab, Lep-etu,
nolatrexed, azd2171, batabulin, ofatumumab, zanolimumab,
edotecarin, tetrandrine, rubitecan, tesmilifene, oblimersen,
ticilimumab, ipilimumab, gossypol, Bio 111, 131-I-TM-601, ALT-110,
BIO 140, CC 8490, cilengitide, gimatecan, IL13-PE38QQR, INO 1001,
IPdR.sub.1 KRX-0402, lucanthone, LY317615, neuradiab, vitespan, Rta
744, Sdx 102, talampanel, atrasentan, Xr 311, romidepsin,
ADS-100380, sunitinib, 5-fluorouracil, vorinostat, etoposide,
gemcitabine, doxorubicin, liposomal doxorubicin,
5'-deoxy-5-fluorouridine, vincristine, temozolomide, ZK-304709,
seliciclib; PD0325901, AZD-6244, capecitabine, L-Glutamic acid,
N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]-
benzoyl]-, disodium salt, heptahydrate, camptothecin, PEG-labeled
irinotecan, tamoxifen, toremifene citrate, anastrazole, exemestane,
letrozole, DES(diethylstilbestrol), estradiol, estrogen, conjugated
estrogen, bevacizumab, IMC-1C11, CHIR-258);
3-[5-(methylsulfonylpiperadinemethyl)-indolyl-quinolone, vatalanib,
AG-013736, AVE-0005, goserelin acetate, leuprolide acetate,
triptorelin pamoate, medroxyprogesterone acetate,
hydroxyprogesterone caproate, megestrol acetate, raloxifene,
bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714;
TAK-165, HKI-272, erlotinib, lapatanib, canertinib, ABX-EGF
antibody, erbitux, EKB-569, PKI-166, GW-572016, Ionafarnib,
BMS-214662, tipifarnib; amifostine, NVP-LAQ824, suberoyl analide
hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248,
sorafenib, KRN951, aminoglutethimide, arnsacrine, anagrelide,
L-asparaginase, Bacillus Calmette-Guerin (BCG) vaccine, adriamycin,
bleomycin, buserelin, busulfan, carboplatin, carmustine,
chlorambucil, cisplatin, cladribine, clodronate, cyproterone,
cytarabine, dacarbazine, dactinomycin, daunorubicin,
diethylstilbestrol, epirubicin, fludarabine, fludrocortisone,
fluoxymesterone, flutamide, gleevec, gemcitabine, hydroxyurea,
idarubicin, ifosfamide, imatinib, leuprolide, levamisole,
lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna,
methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide,
octreotide, oxaliplatin, pamidronate, pentostatin, plicamycin,
porfimer, procarbazine, raltitrexed, rituximab, streptozocin,
teniposide, testosterone, thalidomide, thioguanine, thiotepa,
tretinoin, vindesine, 13-cis-retinoic acid, phenylalanine mustard,
uracil mustard, estramustine, altretamine, floxuridine,
5-deooxyuridine, cytosine arabinoside, 6-mecaptopurine,
deoxycoformycin, calcitriol, valrubicin, mithramycin, vinblastine,
vinorelbine, topotecan, razoxin, marimastat, COL-3, neovastat,
BMS-275291, squalamine, endostatin, SU5416, SU6668, EMD121974,
interleukin-12, IM862, angiostatin, vitaxin, droloxifene,
idoxyfene, spironolactone, finasteride, cimitidine, trastuzumab,
denileukin diftitox, gefitinib, bortezimib, paclitaxel,
cremophor-free paclitaxel, docetaxel, epithilone B, BMS-247550,
BMS-310705, droloxifene, 4-hydroxytamoxifen, pipendoxifene,
ERA-923, arzoxifene, fulvestrant, acolbifene, lasofoxifene,
idoxifene, TSE-424, HMR-3339, ZK186619, topotecan, PTK787/ZK
222584, VX-745, PD 184352, rapamycin,
40-O-(2-hydroxyethyl)-rapamycin, temsirolimus, AP-23573, RAD001,
ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646,
wortmannin, ZM336372, L-779,450, PEG-filgrastim, darbepoetin,
erythropoietin, granulocyte colony-stimulating factor,
zolendronate, prednisone, cetuximab, granulocyte macrophage
colony-stimulating factor, histrelin, pegylated interferon alfa-2a,
interferon alfa-2a, pegylated interferon alfa-2b, interferon
alfa-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab,
hydrocortisone, interleukin-11, dexrazoxane, alemtuzumab,
all-transretinoic acid, ketoconazole, interleukin-2, megestrol,
immune globulin, nitrogen mustard, methylprednisolone, ibritgumomab
tiuxetan, androgens, decitabine, hexamethylmelamine, bexarotene,
tositumomab, arsenic trioxide, cortisone, editronate, mitotane,
cyclosporine, liposomal daunorubicin, Edwina-asparaginase,
strontium 89, casopitant, netupitant, an NK-1 receptor antagonist,
palonosetron, aprepitant, diphenhydramine, hydroxyzine,
metoclopramide, lorazepam, alprazolam, haloperidol, droperidol,
dronabinol, dexamethasone, methylprednisolone, prochlorperazine,
granisetron, ondansetron, dolasetron, tropisetron, pegfilgrastim,
erythropoietin, epoetin alfa, darbepoetin alfa and mixtures
thereof.
[1360] The term "anti-HIV agent" or "additional anti-HIV agent"
includes, for example, nucleoside reverse transcriptase inhibitors
(NRTI), other non-nucloeoside reverse transcriptase inhibitors
(i.e., those which are not representative of the present
disclosure), protease inhibitors, fusion inhibitors, among others,
exemplary compounds of which may include, for example, 3TC
(Lamivudine), AZT (Zidovudine), (-)-FTC, ddI (Didanosine), ddC
(zalcitabine), abacavir (ABC), tenofovir (PMPA), D-D4FC (Reverset),
D4T (Stavudine), Racivir, L-FddC, L-FD4C, NVP (Nevirapine), DLV
(Delavirdine), EFV (Efavirenz), SQVM (Saquinavir mesylate), RTV
(Ritonavir), IDV (Indinavir), SQV (Saquinavir), NFV (Nelfinavir),
APV (Amprenavir), LPV (Lopinavir), fusion inhibitors such as T20,
among others, fuseon and mixtures thereof, including anti-HIV
compounds presently in clinical trials or in development.
[1361] Other anti-HIV agents which may be used in coadministration
with compounds according to the present disclosure include, for
example, other NNRTI's (i.e., other than the NNRTI's according to
the present disclosure) may be selected from the group consisting
of nevirapine (BI-R6-587), delavirdine (U-90152S/T), efavirenz
(DMP-266), UC-781
(N-[4-chloro-3-(3-methyl-2-butenyloxy)phenyl]-2methyl3-furancarbothiamide-
), etravirine (TMC125), Trovirdine (Ly300046.HCl), MKC-442
(emivirine, coactinon), HI-236, HI-240, HI-280, HI-281, rilpivirine
(TMC-278), MSC-127, HBY 097, DMP266, Baicalin (TJN-151) ADAM-II
(Methyl
3',3'-dichloro-4',4''-dimethoxy-5',5''-bis(methoxycarbonyl)-6,6-diphenylh-
exenoate), Methyl
3-Bromo-5-(1-5-bromo-4-methoxy-3-(methoxycarbonyl)phenyl)hept-1-enyl)-2-m-
ethoxybenzoate (Alkenyldiarylmethane analog, Adam analog),
(5-chloro-3-(phenylsulfinyl)-2'-indolecarboxamide), AAP-BHAP
(U-104489 or PNU-104489), Capravirine (AG-1549, S-1153), atevirdine
(U-87201E), aurin tricarboxylic acid (SD-095345),
1-[(6-cyano-2-indolyl)carbonyl]-4-[3-(isopropylamino)-2-pyridinyl]piperaz-
ine,
1-[5-[[N-(methyl)methylsulfonylamino]-2-indolylcarbonyl-4-[3-(isoprop-
ylamino)-2-pyridinyl]piperazine,
1-[3-(Ethylamino)-2-[pyridinyl]-4-[(5-hydroxy-2-indolyl)carbonyl]piperazi-
ne,
1-[(6-Formyl-2-indolyl)carbonyl]-4-[3-(isopropylamino)-2-pyridinyl]pip-
erazine,
1-[[5-(Methylsulfonyloxy)-2-indoyly)carbonyl]-4-[3-(isopropylamin-
o)-2-pyridinyl]piperazine, U88204E, Bis(2-nitrophenyl)sulfone (NSC
633001), Calanolide A (NSC675451), Calanolide B,
6-Benzyl-5-methyl-2-(cyclohexyloxy)pyrimidin-4-one (DABO-546), DPC
961, E-EBU, E-EBU-dm, E-EPSeU, E-EPU, Foscarnet (Foscavir), HEPT
(1-[(2-Hydroxyethoxy)methyl]-6-(phenylthio)thymine), HEPT-M
(1-[(2-Hydroxyethoxy)methyl]-6-(3-methylphenyl)thio)thymine),
HEPT-S (1-[(2-Hydroxyethoxy)methyl]-6-(phenylthio)-2-thiothymine),
Inophyllum P, L-737,126, Michellamine A (NSC650898), Michellamine B
(NSC649324), Michellamine F,
6-(3,5-Dimethylbenzyl)-1-[(2-hydroxyethoxy)methyl]-5-isopropyluracil,
6-(3,5-Dimethylbenzyl)-1-(ethyoxymethyl)-5-isopropyluracil, NPPS,
E-BPTU (NSC 648400), Oltipraz
(4-Methyl-5-(pyrazinyl)-3H-1,2-dithiole-3-thione),
N-{2-(2-Chloro-6-fluorophenethyl]-N'-(2-thiazolyl)thiourea (PETT
Cl, F derivative),
N-{2-(2,6-Difluorophenethyl]-N'-[2-(5-bromopyridyl)]thiourea {PETT
derivative),
N-{2-(2,6-Difluorophenethyl]-N'-[2-(5-methylpyridyl)]thiourea {PETT
Pyridyl derivative),
N-[2-(3-Fluorofuranyl)ethyl]-N'-[2-(5-chloropyridyl)]thiourea,
N-[2-(2-Fluoro-6-ethoxyphenethyl)]-N'-[2-(5-bromopyridyl)]thiourea,
N-(2-Phenethyl)-N'-(2-thiazolyl)thiourea (LY-73497), L-697,639,
L-697,593, L-697,661,
3-[2-(4,7-Difluorobenzoxazol-2-yl)ethyl}-5-ethyl-6-methyl(pypridin-2(1H)--
thione (2-Pyridinone Derivative),
3-[[(2-Methoxy-5,6-dimethyl-3-pyridyl)methyl]amine]-5-ethyl-6-methyl(pypr-
idin-2(1H)-thione, R82150, R82913, R87232, R88703, R89439
(Loviride), R90385, S-2720, Suramin Sodium, TBZ
(Thiazolobenzimidazole, NSC 625487), Thiazoloisoindol-5-one,
(+)(R)-9b-(3,5-Dimethylphenyl-2,3-dihydrothiazolo[2,3-a]isoindol-5(9bH)-o-
ne, Tivirapine (R86183), UC-38 and UC-84, among others.
[1362] The term "pharmaceutically acceptable salt" is used
throughout the specification to describe, where applicable, a salt
form of one or more of the compounds described herein which are
presented to increase the solubility of the compound in the gastric
juices of the patient's gastrointestinal tract in order to promote
dissolution and the bioavailability of the compounds.
Pharmaceutically acceptable salts include those derived from
pharmaceutically acceptable inorganic or organic bases and acids,
where applicable. Suitable salts include those derived from alkali
metals such as potassium and sodium, alkaline earth metals such as
calcium, magnesium and ammonium salts, among numerous other acids
and bases well known in the pharmaceutical art. Sodium and
potassium salts are particularly preferred as neutralization salts
of the phosphates according to the present disclosure.
[1363] The term "pharmaceutically acceptable derivative" is used
throughout the specification to describe any pharmaceutically
acceptable prodrug form (such as an ester, amide other prodrug
group), which, upon administration to a patient, provides directly
or indirectly the present compound or an active metabolite of the
present compound.
EXAMPLES
[1364] General Synthetic Approach
[1365] The synthetic realization and optimization of the
bifunctional molecules as described herein may be approached in a
step-wise or modular fashion. For example, identification of
compounds that bind to the target molecules can involve high or
medium throughput screening campaigns if no suitable ligands are
immediately available. It is not unusual for initial ligands to
require iterative design and optimization cycles to improve
suboptimal aspects as identified by data from suitable in vitro and
pharmacological and/or ADMET assays. Part of the optimization/SAR
campaign would be to probe positions of the ligand that are
tolerant of substitution and that might be suitable places on which
to attach the linker chemistry previously referred to herein. Where
crystallographic or NMR structural data are available, these can be
used to focus such a synthetic effort.
[1366] In a very analogous way one can identify and optimize
ligands for an E3 Ligase, i.e. ULMs/ILMs/VLMs/CLMs/ILMs.
[1367] With PTMs and ULMs (e.g. ILMs, VLMs, CLMs, and/or ILMs) in
hand, one skilled in the art can use known synthetic methods for
their combination with or without a linker moiety. Linker moieties
can be synthesized with a range of compositions, lengths and
flexibility and functionalized such that the PTM and ULM groups can
be attached sequentially to distal ends of the linker. Thus a
library of bifunctional molecules can be realized and profiled in
in vitro and in vivo pharmacological and ADMET/PK studies. As with
the PTM and ULM groups, the final bifunctional molecules can be
subject to iterative design and optimization cycles in order to
identify molecules with desirable properties.
[1368] In some instances, protecting group strategies and/or
functional group interconversions (FGIs) may be required to
facilitate the preparation of the desired materials. Such chemical
processes are well known to the synthetic organic chemist and many
of these may be found in texts such as "Greene's Protective Groups
in Organic Synthesis" Peter G. M. Wuts and Theodora W. Greene
(Wiley), and "Organic Synthesis: The Disconnection Approach" Stuart
Warren and Paul Wyatt (Wiley).
[1369] PTM embodiments of the current invention can be prepared
according to the synthetic routes previously described in the
literature and/or detailed in schemes 1-6 below. These routes can
be modified and adapted to the synthesis of the particular PTM
embodiment using general methods known to those skilled in the art.
In particular, synthetic approaches to the PTMs represented by the
general formula I have been previously described (see, for example,
Barker, A. J. et al. WO199730034 and Barker, A. J. et al.
Bioorganic and Medicinal Chemistry Letters 2001, 11(14), 1911-1914)
and can be generalized as shown in Scheme 1 and 2, where R.sup.22'
is a synthetic precursor of the targeted substitution R.sup.22 into
which in can be converted using general synthetic methods known to
those skilled in the art.
Experimental Procedures
Synthesis of Example 1
(2S,4R)-1-((S)-2-(tert-butyl)-14-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrro-
lo[2,3-d]pyrimidin-6-yl)benzyl)piperazin-1-yl)-4-oxo-6,9,12-trioxa-3-azate-
tradecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-car-
boxamide
##STR00518##
[1371] Synthetic Scheme:
##STR00519## ##STR00520##
1. Step--Synthesis of tert-Butyl
2-(2-(2-(2-((methylsulfonyl)oxy)ethoxy)ethoxy)ethoxy)acetate
##STR00521##
[1373] To a solution of tert-butyl
2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)acetate (300.0 mg, 1.13
mmol) and TEA (344.5 mg, 3.40 mmol) in DCM (5 mL) was added MsCl
(195.8 mg, 1.70 mmol) at 0.degree. C. The solution was stirred at
room temperature for 1 h. The mixture was quenched with water and
then extracted with DCM (10 mL.times.3). The combined organic
layers were washed with brine (15 mL.times.3). The organic phase
was dried over anhydrous sodium sulfate, filtered and concentrated
under reduced pressure to afford crude title compound tert-Butyl
2-(2-(2-(2-((methylsulfonyl)oxy)ethoxy)ethoxy)ethoxy)acetate (350.2
mg, 90.1% yield) as yellow oil, which was used in the next step
without further purification.
[1374] 2. Step to 4. Step--Synthesis of
N-(2-chlorobenzyl)-6-(4-(chloromethyl)phenyl)-7H-pyrrolo[2,3-d]pyrimidin--
4-amine followed procedures analogous to those described by Cai, X.
et al. in WO 2008033747.
##STR00522##
5. Step--Synthesis of tert-Butyl
4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)benzyl)pi-
perazine-1-carboxylate
##STR00523##
[1376] To a solution of
N-(2-chlorobenzyl)-6-(4-(chloromethyl)phenyl)-7H-pyrrolo[2,3-d]pyrimidin--
4-amine (3.93 g, 10.3 mmol) in dioxane (80 mL) was added tert-butyl
piperazine-1-carboxylate (3.92 g, 21.0 mmol). The solution was
stirred at 90.degree. C. for 7 h. The mixture was concentrated
under reduced pressure. The residue was recrystallized from
CH.sub.3OH to give tert-Butyl
4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)benzyl)pi-
perazine-1-carboxylate (4.08 g, 91.9% yield) as alight yellow
solid.
6. Step--Synthesis of
N-(2-Chlorobenzyl)-6-(4-(piperazin-1-ylmethyl)phenyl)-7H-pyrrolo[2,3-d]py-
rimidin-4-amine
##STR00524##
[1378] To a solution of tert-butyl
4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)benzyl)pi-
perazine-1-carboxylate (3.98 g, 7.5 mmol) in DCM (30 mL) was added
trifluoroacetic acid (9.0 mL). The mixture was stirred at RT for 3
h. The solvent was removed under reduced pressure. The residue was
dissolved in water (50 mL). The pH of the solution was adjusted to
9 by solid NaHCO.sub.3. The mixture was extracted with DCM (50
mL.times.3). The combined organic layers were washed with brine.
The organic phase was dried over anhydrous sodium sulfate, filtered
and evaporated under reduced pressure to afford crude product
N-(2-Chlorobenzyl)-6-(4-(piperazin-1-ylmethyl)phenyl)-7H-pyrrolo[2,3-d]py-
rimidin-4-amine (2.0 g, 61.9% yield), which was used in the next
step without further purification.
7. Step--Synthesis of
2-(2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6--
yl)benzyl)piperazin-1-yl)ethoxy)ethoxy)ethoxy)acetic acid
##STR00525##
[1380] To a solution of tert-butyl
2-(2-(2-(2-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6--
yl)benzyl)piperazin-1-yl)ethoxy)ethoxy)ethoxy)acetate (136.2 mg,
0.20 mmol) in dioxane (2.0 mL) was added dioxane/HCl (0.5 mL). The
mixture was stirred at RT for 1.5 h. The solvent was removed in
vacuo to afford crude title product
2-(2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6--
yl)benzyl)piperazin-1-yl)ethoxy)ethoxy)ethoxy)acetic acid (121.5
mg), which was used in the next step without further
purification.
8. Step--Synthesis of
(2S,4R)-1-((S)-2-(tert-butyl)-14-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrr-
olo[2,3-d]pyrimidin-6-yl)benzyl)piperazin-1-yl)-4-oxo-6,9,12-trioxa-3-azat-
etradecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-ca-
rboxamide
##STR00526##
[1382] To a solution of
2-(2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6--
yl)benzyl)piperazin-1-yl)ethoxy)ethoxy)ethoxy)acetic acid (121.5
mg, 0.19 mmol) and
(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4--
methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (87.9 mg, 0.19
mmol) in dry DMF (6.0 mL) were added DIEA (76.7 mg, 0.57 mmol) and
PyBop (197.8 mg, 0.38 mmol). The mixture was stirred at RT for 1.5
h. The solution was quenched with water (20 mL) and then extracted
with EA (20 mL.times.3). The combined organic layers were washed
with brine. The organic phase was dried over anhydrous sodium
sulfate, filtered and evaporated in vacuo. The residue was purified
by silica gel column chromatography (dichloromethane/methanol=25/1)
to afford crude title product (60.2 mg). Then it was further
purified by prep-HPLC to afford the title product of
(2S,4R)-1-((S)-2-(tert-butyl)-14-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrr-
olo[2,3-d]pyrimidin-6-yl)benzyl)piperazin-1-yl)-4-oxo-6,9,12-trioxa-3-azat-
etradecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-ca-
rboxamide (19.0 mg, 9.5% yield).
[1383] .sup.1H NMR (400 MHz, MeOD): .delta. 8.85 (s, 1H), 8.10 (s,
1H), 7.74 (d, J=8.0 Hz, 2H), 7.37-7.44 (m, 8H), 7.23-7.25 (m, 2H),
6.92 (s, 1H), 4.69 (s, 1H), 4.48-4.59 (m, 5H), 4.30 (s, 1H), 4.00
(d, J=5.60 Hz, 2H), 3.87 (d, J=11.08 Hz, 1H), 3.80 (d, J=3.80 Hz,
1H), 3.63-3.68 (m, 12H), 2.90 (s, 4H), 2.65 (s, 3H), 2.45 (s, 3H),
2.22 (m, 1H), 2.21 (m, 1H), 1.03 (s, 9H).
Synthesis of Example 8
4-((17-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)be-
nzyl)piperazin-1-yl)-3,6,9,12,15-pentaoxaheptadecyl)amino)-2-(2,6-dioxopip-
eridin-3-yl)isoindoline-1,3-dione
##STR00527##
[1385] Synthetic Scheme:
##STR00528##
1. Step--Synthesis of
2-(2,6-Dioxopiperidin-3-yl)-4-((17-hydroxy-3,6,9,12,15-pentaoxaheptadecyl-
)amino)isoindoline-1,3-dione
##STR00529##
[1387] To a solution of
17-amino-3,6,9,12,15-pentaoxaheptadecan-1-ol hydrochloride (2.00 g,
7.1 mmol) and
2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (2.94 g,
10.65 mmol) in NMP (10 mL) was added DIEA (3.67 g, 28.4 mmol). The
solution was stirred at 90.degree. C. for 2.5 h. Then it was cooled
to rt and quenched with water (20 mL). The mixture was extracted
with DCM (50 mL.times.3). The combined organic layers were dried
over anhydrous sodium sulfate, filtered and concentrated under
vacuum. The residue was purified by silica gel column
chromatography (DCM/MeOH=25/1) to afford the title compound
2-(2,6-Dioxopiperidin-3-yl)-4-((17-hydroxy-3,6,9,12,15-pentaoxah-
eptadecyl)amino)isoindoline-1,3-dione (300.2 mg, 9.9% yield).
2. Step--Synthesis of
17-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12-
,15-pentaoxaheptadecanal
##STR00530##
[1389] To a solution of
2-(2,6-Dioxopiperidin-3-yl)-4-((17-hydroxy-3,6,9,12,15-pentaoxaheptadecyl-
)amino)isoindoline-1,3-dione (300.0 mg, 0.56 mmol) in CH.sub.3CN
(15.0 mL) was added IBX (234.4 mg, 0.84 mmol). The mixture was
stirred at 80.degree. C. for 2 h. Then it was filtered through
Celite and the filtrate was concentrated under reduced pressure to
afford the crude title product
17-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12-
,15-pentaoxaheptadecanal (230.6 mg, 77.2% yield), which was used in
the next reaction without further purification.
3. Step--Synthesis of
4-((17-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)b-
enzyl)piperazin-1-yl)-3,6,9,12,15-pentaoxaheptadecyl)amino)-2-(2,6-dioxopi-
peridin-3-yl)isoindoline-1,3-dione
##STR00531##
[1391] To a solution of
17-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12-
,15-pentaoxaheptadecanal (150.0 mg, 0.33 mmol) and
N-(2-chlorobenzyl)-6-(4-(piperazin-1-ylmethyl)phenyl)-7H-pyrrolo[2,3-d]py-
rimidin-4-amine (145.0 mg, 0.33 mmol) in MeOH (10 mL) were added
NaCNBH.sub.3(98.0 mg, 1.0 mmol) and two drops of AcOH. The mixture
was stirred at rt for 16 h. Then the solvent was removed in vacuo.
The residue was purified by prep-HPLC to afford the title compound
4-((17-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)b-
enzyl)piperazin-1-yl)-3,6,9,12,15-pentaoxaheptadecyl)amino)-2-(2,6-dioxopi-
peridin-3-yl)isoindoline-1,3-dione (45.9 mg, 17.2% yield).
[1392] .sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 8.26 (s, 1H),
7.77 (d, J=7.6 Hz, 2H), 7.47-7.51 (m, 5H), 7.36-7.39 (m, 2H), 7.15
(s, 1H), 6.99-7.02 (m, 2H), 5.00-5.04 (m, 1H), 4.93 (s, 2H), 3.81
(s, 4H), 3.42-3.69 (m, 24H), 2.69-3.00 (m, 8H), 2.03-2.09 (m, 1H),
1.25-1.32 (m, 1H).
Synthesis of Example 10
##STR00532##
[1393]
4-((2-(2-(2-(2-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrim-
idin-6-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethyl)amino)-2-(2,6-dioxopiperidin--
3-yl)isoindoline-1,3-dione
[1394] Synthetic Scheme:
##STR00533##
1. Step--Synthesis of
6-(4-(2-(2-(2-(2-(N,N-di-tert-butoxycarbonyl-amino)ethoxy)ethoxy)ethoxy)e-
thoxy)phenyl)-N-(2-chlorobenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
##STR00534##
[1396] To a solution of
2-(2-(2-(2-(N,N-di-tert-butoxycarbonyl-amino)aminoethoxy)ethoxy)ethoxy)et-
hyl methanesulfonate (490 mg, 1.04 mmol) in DMF (10 mL) and
4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)phenol
(364 mg, 1.04 mmol) was added K.sub.2CO.sub.3 (430 mg, 3.12 mmol)
at 25.degree. C. The resulting solution was stirred at 70.degree.
C. for 16 h. The resulting solution was cooled to 20.degree. C. The
mixture was diluted with H.sub.2O (40 mL). The mixture was
extracted with EtOAc (40 mL.times.2). The combined organic layers
were dried over anhydrous sodium sulfate and concentration. The
residue was purified with silica gel column to afford the title
compound
6-(4-(2-(2-(2-(2-(N,N-di-tert-butoxycarbonyl-amino)ethoxy)ethoxy)ethoxy)e-
thoxy)phenyl)-N-(2-chlorobenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(280 mg, 37.1% yield).
2. Step and 3. Step--Synthesis of
4-((2-(2-(2-(2-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-
-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethyl)amino)-2-(2,6-dioxopiperidin-3-yl)i-
soindoline-1,3-dione
##STR00535##
[1398] To a solution of
6-(4-(2-(2-(2-(2-(N,N-di-tert-butoxycarbonyl-amino)ethoxy)ethoxy)ethoxy)e-
thoxy)phenyl)-N-(2-chlorobenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(280 mg, 0.38 mmol) in dioxane (5 mL) was added HCl (g)/dioxane (2
mL) at 0.degree. C., and then the reaction was stirred at
25.degree. C. for 2 h. The solvent was removed under vacuum. The
residue was dissolved into NMP (5 mL). To the above mixture were
added 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione
(334.6 mg, 1.16 mmol) and DIPEA (245 mg, 1.9 mmol) at 25.degree. C.
subsequently. The reaction was microwave irradiated to 150.degree.
C. for 20 min. Then it was cooled to RT and quenched by addition of
water (20 mL). The mixture was extracted with DCM (50 mL.times.3).
The combined organic layers were dried over anhydrous sodium
sulfate and concentrated under vacuum. The residue was purified
with silica gel column to afford the crude product, and it was
purified again by prep-HPLC to afford the title compound
4-((2-(2-(2-(2-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-
-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethyl)amino)-2-(2,6-dioxopiperidin-3-yl)i-
soindoline-1,3-dione (28 mg, 9.2% yield).
[1399] .sup.1HNMR (400 MHz, CDCl.sub.3): .delta. 11.38 (br, 1H),
10.29 (s, 1H), 8.34 (s, 1H), 7.49-7.53 (m, 3H), 7.39-7.44 (m, 2H),
7.22-7.24 (m, 2H), 7.05 (d, J=7.2 Hz, 1H), 6.95 (d, J=8.4 Hz, 1H),
6.88 (d, J=8.8 Hz, 2H), 6.47 (s, 1H), 6.39-6.42 (m, 1H), 4.95-4.98
(m, 3H), 4.12 (t, J=5.2 Hz, 2H), 3.85 (t, J=5.2 Hz, 2H), 3.67-3.73
(m, 10H), 3.42-3.46 (m, 2H), 2.65-2.91 (m, 3H), 2.09-2.15 (m,
2H).
Synthesis of Example 14
(2S,4R)-1-((S)-2-(tert-butyl)-14-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[-
2,3-d]pyrimidin-6-yl)phenoxy)-4-oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-hy-
droxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
##STR00536##
[1401] Synthetic Scheme:
##STR00537##
[1402] 1. Step to 2. Step--Synthesis of
4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)phenol
starting from
4-chloro-6-(4-methoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidine using
procedures analogous to those described by Kaspersen, S. et al. in
Bioorganic Chemistry 2012, 44, 35-41.
3. Step--Synthesis of tert-Butyl
2-(2-(2-(2-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
phenoxy)ethoxy)ethoxy)ethoxy)acetate
##STR00538##
[1404] To a solution of
4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)phenol
(500 mg, 1.16 mmol) in DMF (10 mL) were added tert-butyl
2-(2-(2-(2-((methylsulfonyl)oxy)ethoxy)ethoxy)ethoxy)acetate (515
mg, 1.5 mmol) and K.sub.2CO.sub.3 (480 mg, 3.48 mmol). The solution
was stirred at 80.degree. C. for 20 h. The mixture was extracted
with EtOAc (20 mL.times.3). The combined organic layers were washed
with brine. The organic layer was dried over anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The
residue was purified by silica gel column chromatography
(PE/EA=1/1) to afford the title compound tert-Butyl
2-(2-(2-(2-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
phenoxy)ethoxy)ethoxy)ethoxy)acetate (320 mg) as a yellow
solid.
4. Step and 5. Step--Synthesis of
(2S,4R)-1-((S)-2-(tert-butyl)-14-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo-
[2,3-d]pyrimidin-6-yl)phenoxy)-4-oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-h-
ydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
##STR00539##
[1406] tert-Butyl
2-(2-(2-(2-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
phenoxy)ethoxy)ethoxy)ethoxy)acetate was converted to the final
compound,
(2S,4R)-1-((S)-2-(tert-butyl)-14-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo-
[2,3-d]pyrimidin-6-yl)phenoxy)-4-oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-h-
ydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
using as described for the example 1 above.
Synthesis of Example 17
5-(2-(2-(2-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-y-
l)benzyl)piperazin-1-yl)ethoxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)i-
soindoline-1,3-dione
##STR00540##
[1408] Synthetic Scheme:
##STR00541##
1. Step--Synthesis of
2-(2,6-Dioxopiperidin-3-yl)-5-(2-(2-(2-hydroxyethoxy)ethoxy)exthoxy)isoin-
doline-1,3-dione
##STR00542##
[1410] To a solution of
2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (500 mg,
1.82 mmol) in DMF (10 mL) were added K.sub.2CO.sub.3 (756 mg, 5.47
mmol) and 2-(2-(2-hydroxyethoxy)ethoxy)ethyl
4-methyl-benzenesulfonate (832 mg, 2.73 mmol) at 25.degree. C. The
resulting solution was stirred at 70.degree. C. for 5 h. After
cooling to rt, the reaction was quenched with H.sub.2O (10 mL), and
the mixture was extracted with EtOAc (10 mL.times.2). The combined
organic layers were dried over anhydrous sodium sulfate and
concentrated. The residue was purified with silica gel column to
afford the title product
2-(2,6-Dioxopiperidin-3-yl)-5-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)isoind-
oline-1,3-dione (95 mg, 13% yield).
2. Step--Synthesis of
2-(2-(2-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy-
)ethoxy)acetaldehyde
##STR00543##
[1412] To a solution of
2-(2,6-dioxopiperidin-3-yl)-5-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)isoind-
oline-1,3-dione (95 mg, 0.23 mmol) in CH.sub.3CN (5 mL) was added
IBX (130 mg, 0.46 mmol) at 25.degree. C. The reaction was stirred
at 80.degree. C. for 2 h. After cooling to rt, the mixture was
filtered through Celite, and the filtrate was concentrated to
afford title product
2-(2-(2-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy-
)ethoxy)acetaldehyde (90 mg, crude), which was used in next step
without further purification.
3. Step--Synthesis of
5-(2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6--
yl)benzyl)piperazin-1-yl)ethoxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)-
isoindoline-1,3-dione
##STR00544##
[1414] To a solution of
2-(2-(2-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy-
)ethoxy)acetaldehyde (90 mg, 0.15 mmol) and
N-(2-chlorobenzyl)-6-(4-(piperazin-1-ylmethyl)phenyl)-7H-pyrrolo[2,3-d]py-
rimidin-4-amine (79.2 mg, 0.18 mmol) in DMSO/MeOH (2 mL/2 mL) was
added NaBH.sub.3CN (47.9 mg, 0.76 mmol) at 10.degree. C. The
resulting mixture was stirred at 15.degree. C. for 0.5 h. The
mixture was quenched with H.sub.2O (10 mL), and the mixture was
extracted with EtOAc (10 mL.times.2). The combined organic layers
were washed with brine (10 mL.times.2), dried over anhydrous sodium
sulfate and concentrated. The residue was purified with silica gel
column and prep-HPLC to afford the title compound
5-(2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6--
yl)benzyl)piperazin-1-yl)ethoxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)-
isoindoline-1,3-dione (21 mg, 11% yield).
[1415] .sup.1H NMR (400 MHz, DMSO-d6): .delta. 12.09 (s, 1H), 11.14
(s, 1H), 8.06-8.11 (m, 2H), 7.74-7.85 (m, 3H), 7.30-7.46 (m, 8H),
7.01 (s, 1H), 5.13 (d, J=7.6 Hz, 1H), 4.79 (d, J=4.4 Hz, 2H), 4.43
(s, 2H), 3.79 (s, 2H), 3.38-3.59 (m, 10H), 2.06-2.62 (m, 10H), 1.91
(s, 2H).
Synthesis of Example 18
(2S,4R)-1-((S)-2-(tert-butyl)-20-((4-((3-chloro-4-fluorophenyl)amino)-7-me-
thoxyquinazolin-6-yl)oxy)-4-oxo-6,9,12,15,18-pentaoxa-3-azaicosanoyl)-4-hy-
droxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
##STR00545##
##STR00546##
[1416] Experimental Details
1. Step--Synthesis of 5-hydroxy-4-methoxy-2-nitrobenzoic acid
##STR00547##
[1418] Into a 3-L round-bottom flask, was placed a solution of
methyl 4,5-dimethoxy-2-nitrobenzoate (45 g, 186.57 mmol, 1.00
equiv) in water (800 mL) and DME (200 mL), potassium hydroxide (72
g, 1.28 mol, 7.00 equiv). The resulting solution was refluxed for 1
day. The resulting mixture was washed with 2.times.200 mL of
hexane. The pH value of the solution was adjusted to 5 with
hydrogen chloride (6 mol/L). The solids were collected by
filtration. The resulting mixture was concentrated under vacuum.
This resulted in 40 g of 5-hydroxy-4-methoxy-2-nitrobenzoic acid as
a yellow solid. LC-MS: (ES, m/z): 214 [M+H].sup.+ Retention time:
0.217 min
2. Step--Synthesis of 2-amino-5-hydroxy-4-methoxybenzoic acid
##STR00548##
[1420] Into a 1-L round-bottom flask, was placed
5-hydroxy-4-methoxy-2-nitrobenzoic acid (41.2 g, 193.30 mmol, 1.00
equiv), 10% Palladium carbon (5 g, 0.10 equiv), methanol (500 mL).
H.sub.2 was introduced into the reaction mixture. The resulting
solution was stirred for overnight at room temperature under
H.sub.2 atmosphere. The solids were filtered out. The resulting
mixture was concentrated under vacuum. This resulted in 21.3 g
(60%) of 2-amino-5-hydroxy-4-methoxybenzoic acid as a yellow solid.
LC-MS: (ES, m/z): 184 [M+H].sup.+ Retention time: 0.356 min
3. Step--Synthesis of 7-methoxyquinazoline-4,6-diol
##STR00549##
[1422] Into a 500-mL round-bottom flask, was placed a solution of
2-amino-5-hydroxy-4-methoxybenzoic acid (21.3 g, 116.29 mmol, 1.00
equiv) in MeO(CH.sub.2).sub.2OH (200 mL) and methanimidamide
monoacetate (12.6 g, 122.33 mmol, 1.10 equiv). The resulting
solution was stirred for 30 min at 140.degree. C. The resulting
mixture was concentrated under vacuum. This resulted in 20 g (89%)
of 7-methoxyquinazoline-4,6-diol as a black solid.
[1423] LC-MS: (ES, m/z): 193 [M+H].sup.+ Retention time: 0.959
min
4. Step--Synthesis of 4-hydroxy-7-methoxyquinazolin-6-yl
acetate
##STR00550##
[1425] Into a 500-mL round-bottom flask, was placed
7-methoxyquinazoline-4,6-diol (12 g, 62.44 mmol, 1.00 equiv),
Ac.sub.2O (200 mL), pyridine (20 mL). The resulting solution was
stirred for 3 h at 100.degree. C. The resulting mixture was
concentrated under vacuum. The residue was purified on combi-flash
with MeOH/DCM (1:100-1:10). This resulted in 8.5 g (58%) of
4-hydroxy-7-methoxyquinazolin-6-yl acetate as a brown solid. LC-MS:
(ES, m/z): 235 [M+H].sup.+ Retention time: 1.251 min
5. Step--Synthesis of 4-chloro-7-methoxyquinazolin-6-yl acetate
##STR00551##
[1427] Into a 250-mL round-bottom flask, was placed
4-hydroxy-7-methoxyquinazolin-6-yl acetate (8.5 g, 36.29 mmol, 1.00
equiv), thionyl chloride (100 mL), N,N-dimethylformamide (1 mL).
The resulting solution was stirred for 1.5 h at 85.degree. C. The
resulting solution was evaporated in vacuum, extracted with
3.times.100 mL of ethyl acetate and the organic layers combined and
dried in an oven under reduced pressure, concentrated under vacuum.
The residue was purified on combi-flash with MeOH/DCM (1:100-1:10).
This resulted in 9.1 g (99%) of 4-chloro-7-methoxyquinazolin-6-yl
acetate as a brown solid. LC-MS: (ES, m/z): 253 [M+H].sup.+
Retention time: 0.744 min
6. Step--Synthesis of
4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
acetate
##STR00552##
[1429] Into a 500-mL round-bottom flask, was placed
4-chloro-7-methoxyquinazolin-6-yl acetate (9.1 g, 36.02 mmol, 1.00
equiv), 3-chloro-4-fluoroaniline (5.23 g, 35.93 mmol, 1.00 equiv)
in propan-2-ol (200 mL). The resulting solution was refluxed for 3
hours. The reaction mixture was cooled to room temperature. The
solids were collected by filtration. This resulted in 10.1 g (78%)
of 4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
acetate as a brown solid. LC-MS: (ES, m/z): 362 [M+H].sup.+
Retention time: 0.681 min
7. Step--Synthesis of
4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-ol
##STR00553##
[1431] Into a 500-mL round-bottom flask, was placed a solution of
4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-yl acetate
(10.1 g, 27.92 mmol, 1.00 equiv) in methanol (200 mL), a solution
of NaOH (4 g, 100.01 mmol, 5.00 equiv) in water (20 mL). The
resulting solution was stirred for overnight at room temperature.
The pH value of the solution was adjusted to 5 with hydrogen
chloride (1 mol/L). The solids were collected by filtration. This
resulted in 7.5 g (84%) of
4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-ol as a
brown solid. LC-MS: (ES, m/z): 320 [M+H].sup.+ Retention time:
0.904 min
[1432] .sup.1H-NMR: (DMSO, ppm): .delta.=11.15-11.12 (s, 1H),
10.59-10.51 (s, 1H), 9.88-9.82 (s, 1H), 8.25-7.99 (m, 2H),
7.86-7.69 (m, 1H), 7.58-7.47 (m, 1H), 7.37-7.33 (s, 1H), 4.06-3.99
(s, 3H).
##STR00554##
Experimental Details
8. Step--Synthesis of ethyl
17-hydroxy-3,6,9,12,15-pentaoxaheptadecanoate
##STR00555##
[1434] Into a 500-mL 3-necked round-bottom flask, was placed
dichloromethane (150 mL), 3,6,9,12-tetraoxatetradecane-1,14-diol
(18 g, 75.54 mmol, 2.00 equiv), BF.sub.3-Et.sub.2O (1 mL). This was
followed by the addition of ethyl 2-diazenylacetate (4.3 g, 37.03
mmol, 1.00 equiv) dropwise with stirring at 0.degree. C. in 1 hr.
The resulting solution was stirred for 2 h at 25.degree. C. The
reaction was then quenched by the addition of 200 mL of water. The
resulting solution was extracted with 150 mL of dichloromethane and
the organic layers combined and concentrated under vacuum. The
residue was applied onto a silica gel column with
dichloromethane/methanol (50:1). This resulted in 1.5 g of ethyl
17-hydroxy-3,6,9,12,15-pentaoxaheptadecanoate as white oil. LC-MS:
(ES, m/z): 325 [M+H].sup.+ Retention time: 1.285 min
9. Step--Synthesis of ethyl
17-[[(4-methylbenzene)sulfonyl]oxy]-3,6,9,12,15-pentaoxaheptadecanoate
##STR00556##
[1436] Into a 100-mL round-bottom flask, was placed ethyl
17-hydroxy-3,6,9,12,15-pentaoxaheptadecanoate (3.2 g, 9.87 mmol,
1.00 equiv), dichloromethane (50 mL), triethylamine (1.52 g, 15.02
mmol, 1.50 equiv), 4-dimethylaminopyridine (183 mg, 1.50 mmol, 0.10
equiv). This was followed by the addition of TsCl (2.09 g, 10.96
mmol, 1.10 equiv) in portions. The resulting solution was stirred
overnight at 25.degree. C. The reaction was then quenched by the
addition of 20 mL of water. The resulting solution was extracted
with 2.times.30 mL of dichloromethane and the organic layers
combined and dried over anhydrous sodium sulfate and concentrated
under vacuum. The residue was applied onto a silica gel column with
dichloromethane/methanol (40:1). This resulted in 1 g (21%) of
ethyl
17-[[(4-methylbenzene)sulfonyl]oxy]-3,6,9,12,15-pentaoxaheptadec-
anoate as light yellow oil. LC-MS: (ES, m/z): 479 [M+H].sup.+
Retention time: 1.480 min
10. Step--Synthesis of ethyl
1-[4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-yl]-1,4,7,10,-
13,16-hexaoxaoctadecan-18-oate
##STR00557##
[1438] Into a 50-mL round-bottom flask purged and maintained with
an inert atmosphere of nitrogen, was placed
4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-ol (100
mg, 0.31 mmol, 1.00 equiv), N,N-dimethylformamide (10 mL),
potassium carbonate (86.4 mg, 0.63 mmol, 2.00 equiv), ethyl
17-[[(4-methylbenzene)sulfonyl]oxy]-3,6,9,12,15-pentaoxaheptadecanoate
(224 mg, 0.47 mmol, 1.50 equiv). The resulting solution was stirred
overnight at 80.degree. C. The reaction was then quenched by the
addition of 10 mL of water. The resulting solution was extracted
with 2.times.10 mL of ethyl acetate and the organic layers combined
and dried over anhydrous sodium sulfate and concentrated under
vacuum. The residue was applied onto a silica gel column with ethyl
acetate/petroleum ether (1:1). This resulted in 157 mg (80%) of
ethyl
1-[4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-yl]-1,4,7,10,-
13,16-hexaoxaoctadecan-18-oate as light yellow oil. LC-MS: (ES,
m/z): 626 [M+H].sup.+ Retention time: 1.279 min
11. Step--Synthesis of
1-[4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-yl]-1,4,7,10,-
13,16-hexaoxaoctadecan-18-oic acid
##STR00558##
[1440] Into a 50-mL round-bottom flask, was placed ethyl
1-[4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-yl]-1,4,7,10,-
13,16-hexaoxaoctadecan-18-oate (158 mg, 0.25 mmol, 1.00 equiv) in
methanol (10 mL), sodium hydroxide (50.3 mg, 1.26 mmol, 5.00 equiv)
in water (1 mL). The resulting solution was stirred for 2 h at
25.degree. C. The resulting mixture was concentrated under vacuum.
The pH value of the solution was adjusted to 5 with hydrogen
chloride (1 mol/L). The solids were collected by filtration. This
resulted in 67 mg (44%) of
1-[4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-yl]-1,4,7,10,-
13,16-hexaoxaoctadecan-18-oic acid as a white solid. LC-MS: (ES,
m/z): 598 [M+H].sup.+ Retention time: 1.192 min
12. Step--Synthesis of
(2S,4R)-4-(tert-butoxy)-1-[(2S)-2-(1-[4-[(3-chloro-4-fluorophenyl)amino]--
7-methoxyquinazolin-6-yl]-1,4,7,10,13,16-hexaoxaoctadecan-18-amido)-3,3-di-
methylbutanoyl]-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-
-2-carboxamide
##STR00559##
[1442] Into a 50-mL round-bottom flask, was placed
1-[4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-yl]-1,4,7,10,-
13,16-hexaoxaoctadecan-18-oic acid (67 mg, 0.11 mmol, 1.00 equiv)
in N,N-dimethylformamide (3 mL), DIEA (28.4 mg, 0.22 mmol, 2.00
equiv), HATU (55.4 mg, 0.15 mmol, 1.30 equiv),
(2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-(tert-butoxy)-N-[[4-(4-me-
thyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (54.5
mg, 0.11 mmol, 1.00 equiv). The resulting solution was stirred
overnight at 25.degree. C. The reaction was then quenched by the
addition of 20 mL of water. The resulting solution was extracted
with 2.times.50 mL of ethyl acetate and the organic layers combined
and dried over anhydrous sodium sulfate and concentrated under
vacuum. The residue was applied onto a silica gel column with
dichloromethane/methanol (20:1). This resulted in 84 mg (70%) of
(2S,4R)-4-(tert-butoxy)-1-[(2S)-2-(1-[4-[(3-chloro-4-fluorophenyl)amino]--
7-methoxyquinazolin-6-yl]-1,4,7,10,13,16-hexaoxaoctadecan-18-amido)-3,3-di-
methylbutanoyl]-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-
-2-carboxamide as a light yellow solid. LC-MS: (ES, m/z): 1066
[M+H].sup.+ Retention time: 1.409 min
13. Step--Synthesis of
(2S,4R)-1-[(2S)-2-(1-[4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazo-
lin-6-yl]-1,4,7,10,13,16-hexaoxaoctadecan-18-amido)-3,3-dimethylbutanoyl]--
4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-car-
boxamide
##STR00560##
[1444] Into a 25-mL round-bottom flask, was placed a solution of
(2S,4R)-4-(tert-butoxy)-1-[(2S)-2-(1-[4-[(3-chloro-4-fluorophenyl)amino]--
7-methoxyquinazolin-6-yl]-1,4,7,10,13,16-hexaoxaoctadecan-18-amido)-3,3-di-
methylbutanoyl]-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-
-2-carboxamide (81 mg, 0.08 mmol, 1.00 equiv) in dichloromethane (5
mL). This was followed by the addition of CF.sub.3COOH (0.5 mL)
dropwise with stirring at 0.degree. C. The resulting solution was
stirred overnight at 25.degree. C. The resulting mixture was
concentrated under vacuum. The crude product was purified by
Prep-HPLC with the following conditions (Prep-HPLC-043): Column,
XBridge Prep C18 OBD Column, 30*50 mm 5 um 13 nm; mobile phase,
WATER with 0.05% TFA and MeCN (35.0% MeCN up to 65.0% in 8 min);
Detector, Waters 2489 254&220 nm. This resulted in 20 mg (23%)
of
(2S,4R)-1-[(2S)-2-(1-[4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquin-
azolin-6-yl]-1,4,7,10,13,16-hexaoxaoctadecan-18-amido)-3,3-dimethylbutanoy-
l]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2--
carboxamide; trifluoroacetic acid as a white solid. LC-MS: (ES,
m/z): 1010 [M+H].sup.+ Retention time: 3.344 min
[1445] .sup.1H-NMR: (CD.sub.3OD, ppm): .delta.=8.89 (s, 1H), 8.46
(s, 1H), 8.02-7.99 (dd, J=4.0 Hz, 5.6 Hz, 1H), 7.75 (s, 1H),
7.69-7.65 (m, 1H), 7.41-7.18 (m, 6H), 4.87 (s, 1H), 4.68-4.49 (m,
3H), 4.36-4.31 (m, 3H), 4.10 (m, 9H), 3.80-3.73 (m, 3H), 3.68-3.60
(m, 13H), 3.30 (s, 3H), 2.47-2.21 (m, 1H), 2.11-2.07 (m, 1H),
1.56-1.54 (m, 1H), 1.29 (s, 1H), 1.02-1.00 (t, J=8.8 Hz, 9H).
Synthesis of Example 39
(2S,4R)-1-((S)-2-(3-(2-((5-((4-((3-chloro-4-fluorophenyl)amino)-7-methoxyq-
uinazolin-6-yl)oxy)pentyl)oxy)ethoxy)propanamido)-3,3-dimethylbutanoyl)-4--
hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
##STR00561##
[1447] Synthetic Scheme:
##STR00562##
1. Step--Synthesis of 5-(Benzyloxy)pentan-1-ol
##STR00563##
[1449] Into a 1000 mL round-bottom flask, was placed pentane-1,
5-diol (30 g, 288.05 mmol, 1.00 equiv), tetrahydrofuran (500 mL).
This was followed by the addition of sodium hydride (13.8 g, 575.00
mmol, 2.00 equiv) in several batches. The mixture was stirred for 1
h at 25.degree. C. To this was added BnBr (58 g, 339.12 mmol, 1.20
equiv) dropwise with stirring. The resulting solution was stirred
overnight at 25.degree. C. The reaction was then quenched by the
addition of 50 mL of water. The resulting solution was extracted
with 3.times.500 mL of ethyl acetate and the organic layers were
combined and dried over anhydrous sodium sulfate. The solids were
filtered out. The resulting mixture was concentrated under vacuum.
The residue was applied onto a silica gel column eluted with ethyl
acetate/petroleum ether (1:5). This resulted in 28 g (50%) of
5-(benzyloxy)pentan-1-ol as colorless oil.
[1450] LC-MS m/z: (ES+) [M+H]+=195; Retention time: 1.01 min;
[1451] 1H NMR (300 MHz, CDCl3, 25.degree. C.): 7.35 (s, 5H), 4.52
(s, 2H), 3.65 (t, 2H), 3.51 (t, 2H), 1.69-1.40 (m, 6H).
2. Step--Synthesis of
2-(2-[[5-(Benzyloxy)pentyl]oxy]ethoxy)oxane
##STR00564##
[1453] Into a 100 mL round-bottom flask, was placed
5-(benzyloxy)pentan-1-ol (3 g, 15.44 mmol, 1.00 equiv), 50% sodium
hydroxide solution (20 mL), 2-(2-bromoethoxy)oxane (12.8 g, 61.22
mmol, 4.00 equiv), Bu4NHSO4 (0.5 g, 0.10 equiv). The resulting
solution was stirred for 12 h at 65.degree. C. The reaction mixture
was cooled. The resulting mixture was washed with 20 mL of water
and 20 mL of brine. The mixture was dried over anhydrous sodium
sulfate. The solids were filtered out. The resulting mixture was
concentrated under vacuum. This resulted in 4 g (80%) of
2-(2-[[5-(benzyloxy)pentyl]oxy]ethoxy)oxane as red oil. LC-MS m/z:
(ES+) [M+H]+=323; Retention time: 1.25 min.
3. Step--Synthesis of 2-[[5-(Benzyloxy)pentyl]oxy]ethan-1-ol
##STR00565##
[1455] Into a 100 mL round-bottom flask, was placed
2-(2-[[5-(benzyloxy)pentyl]oxy]ethoxy)oxane (4 g, 12.41 mmol, 1.00
equiv), methanol (40 mL), hydrogen chloride (2 mL). The resulting
solution was stirred overnight at 50.degree. C. The reaction
mixture was cooled. The resulting mixture was washed with water and
brine. The mixture was dried over anhydrous sodium sulfate. The
solids were filtered out. The resulting mixture was concentrated
under vacuum. The residue was applied onto a silica gel column
eluted with ethyl acetate/petroleum ether (1:2). This resulted in 3
g (100%) of 2-[[5-(benzyloxy)pentyl]oxy]ethan-1-ol as colorless
oil. LC-MS m/z: (ES+) [M+H]+=239; Retention time: 1.12 min.
4. Step--Synthesis of 2-(3-Bromopropoxy)oxane
##STR00566##
[1457] Into a 250 mL round-bottom flask, was placed
3-bromopropan-1-ol (4.75 g, 34.17 mmol, 1.00 equiv),
dichloromethane (100 mL), PPTs (10 mg, 0.04 mmol, 0.10 equiv),
3,4-dihydro-2H-pyran (3.32 g, 39.47 mmol, 1.16 equiv). The
resulting solution was stirred for 5 h at room temperature. The
mixture was dried over anhydrous magnesium sulfate. The solids were
filtered out. The resulting mixture was concentrated under vacuum.
The residue was applied onto a silica gel column eluted with ethyl
acetate/petroleum ether (1:5). This resulted in 5 g (66%) of
2-(3-bromopropoxy)oxane as colorless oil. 1H NMR (300 MHz, CDCl3,
25.degree. C.): 4.62 (t, 1H), 3.95-3.85 (m, 2H), 3.59-3.48 (m, 4H),
2.18-2.10 (m, 2H), 1.90-1.45 (m, 6H).
5. Step--Synthesis of
2-[3-(2-[[5-(benzyloxy)pentyl]oxy]ethoxy)propoxy]oxane
##STR00567##
[1459] Into a 50 mL round-bottom flask, was placed
2-[[5-(benzyloxy)pentyl]oxy]ethan-1-ol (150 mg, 0.63 mmol, 1.00
equiv), 2 mL of 50% NaOH solution, 4 equivalents of
2-(3-bromopropoxy)oxane, and catalytic amount of Bu4NHSO4 (0.1 eq).
The resulting solution was stirred overnight at 65.degree. C. The
reaction mixture was cooled. The resulting solution was extracted
with 3.times.50 mL of ethyl acetate and the organic layers were
combined and dried over anhydrous sodium sulfate and concentrated
under vacuum. The residue was applied onto a silica gel column
eluted with ethyl acetate/petroleum ether (1:2). This resulted in
200 mg (84%) of
2-[3-(2-[[5-(benzyloxy)pentyl]oxy]ethoxy)propoxy]oxane as colorless
oil. LC-MS m/z: (ES+) [M+Na]+=403; Retention time: 1.53 min; 1H NMR
(300 MHz, CDCl3, 25.degree. C.): 7.35 (s, 5H), 4.62 (t, 1H), 4.52
(s, 2H), 3.95-3.85 (m, 4H), 3.59-3.48 (m, 10H), 1.90-1.45 (m,
14H).
6. Step--Synthesis of
5-[2-[3-(Oxan-2-yloxy)propoxy]ethoxy]pentan-1-ol
##STR00568##
[1461] Into a 50 mL round-bottom flask, was placed
2-[3-(2-[[5-(benzyloxy)pentyl]oxy]ethoxy)propoxy]oxane (80 mg, 0.21
mmol, 1.00 equiv), methanol (5 mL), palladium on carbon (200 mg,
0.20 equiv). To this mixture H2(g) was introduced in. The resulting
solution was stirred overnight at room temperature. The solids were
filtered out. The resulting mixture was concentrated under vacuum.
This resulted in 64 mg (crude) of
5-[2-[3-(oxan-2-yloxy)propoxy]ethoxy]pentan-1-ol as colorless oil.
LC-MS m/z: (ES+) [M+H]+=291; Retention time: 1.32 min; 1H NMR (300
MHz, CDCl3, 25.degree. C.): 4.62 (t, 1H), 3.98-3.79 (m, 2H),
3.65-3.47 (m, 8H), 1.90-1.45 (m, 14H).
7. Step--Synthesis of 5-[2-[3-(Oxan-2-yloxy)propoxy]ethoxy]pentyl
4-methylbenzene-1-sulfonate
##STR00569##
[1463] Into a 50 mL round-bottom flask, was placed
5-[2-[3-(oxan-2-yloxy)propoxy]ethoxy]pentan-1-ol (60 mg, 0.21 mmol,
1.00 equiv), dichloromethane (2 mL), triethylamine (47 mg, 0.46
mmol, 3.00 equiv), TsCl (30 mg, 0.16 mmol, 1.50 equiv). The
resulting solution was stirred overnight at room temperature. The
resulting mixture was washed with water and brine. The mixture was
dried over anhydrous sodium sulfate. The solids were filtered out.
The resulting mixture was concentrated under vacuum. This resulted
in 90 mg (98%) of 5-[2-[3-(oxan-2-yloxy)propoxy]ethoxy]pentyl
4-methylbenzene-1-sulfonate as colorless oil. LC-MS m/z: (ES+)
[M+H]+=445, Retention time: 1.25 min.
8. Step--Synthesis of
3-[2-[(5-[[(4-Methylbenzene)sulfonyl]oxy]pentyl)oxy]ethoxy]propan-1-ol
##STR00570##
[1465] Into a 50 mL round-bottom flask, was placed
5-[2-[3-(oxan-2-yloxy)propoxy]ethoxy]pentyl
4-methylbenzene-1-sulfonate (90 mg, 0.20 mmol, 1.00 equiv),
methanol (2 mL), hydrogen chloride (0.5 mL). The resulting solution
was stirred for 2 h at room temperature. The resulting mixture was
concentrated under vacuum. The resulting solution was extracted
with 3.times.20 mL of ethyl acetate and the organic layers were
combined and dried over anhydrous sodium sulfate. The solids were
filtered out. The resulting mixture was concentrated under vacuum.
This resulted in 45 mg (62%) of
3-[2-[(5-[[(4-methylbenzene)sulfonyl]oxy]pentyl)oxy]ethoxy]propan-1-ol
as colorless oil. LC-MS m/z: (ES+) [M+H]+=291; Retention time: 0.93
min; 1H NMR (400 MHz, CDCl3, 25.degree. C.): 7.83 (d, 2H), 7.35 (d,
2H), 4.05 (t, 2H), 3.80 (t, 2H), 3.70 (t, 2H), 3.64 (d, 2H), 3.58
(d, 2H), 3.45 (t, 2H), 2.92 (brs, 1H), 2.47 (s, 3H), 1.91-1.82 (m,
2H), 1.73-1.65 (m, 2H), 1.58-1.52 (m, 2H), 1.45-1.35 (m, 2H).
9. Step--Synthesis of
3-[2-[(5-[[(4-Methylbenzene)sulfonyl]oxy]pentyl)oxy]ethoxy]propanoic
acid
##STR00571##
[1467] Into a 50 mL round-bottom flask, was placed
3-[2-[(5-[[(4-methylbenzene)sulfonyl]oxy]pentyl)oxy]ethoxy]propan-1-ol
(100 mg, 0.28 mmol, 1.00 equiv), acetone (2 mL). To this was added
CrO3 (55 mg, 2.00 equiv), sulfuric acid (0.1 mL), water (0.6 mL)
under ice bath. The resulting solution was stirred for 2 h at
5-10.degree. C. The reaction was then quenched by the addition of
iso-propanol. The resulting solution was extracted with 2.times.10
mL of ethyl acetate and the organic layers were combined and dried
over anhydrous sodium sulfate. The solids were filtered out. The
resulting mixture was concentrated under vacuum. This resulted in
90 mg (87%) of
3-[2-[(5-[[(4-methylbenzene)sulfonyl]oxy]pentyl)oxy]ethoxy]propanoic
acid as colorless oil.
[1468] LC-MS m/z: (ES+) [M+H]+=375; Retention time: 0.92 min.
10. Step--Synthesis of
(2S,4R)-1-[(2S)-3,3-Dimethyl-2-(3-[2-[(5-[[(4-methylbenzene)-sulfonyl]oxy-
]pentyl)oxy]ethoxy]propanamido)butanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thi-
azol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide
##STR00572##
[1470] Into a 50 mL round-bottom flask, was placed
3-[2-[(5-[[(4-methylbenzene)sulfonyl]oxy]pentyl)oxy]ethoxy]propanoic
acid (112 mg, 0.30 mmol, 1.00 equiv),
(2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1-
,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (90 mg,
0.21 mmol, 1.00 equiv), HATU (137 mg, 0.36 mmol, 1.50 equiv),
N,N-dimethylformamide (2 mL), DIEA (124 mg, 0.96 mmol, 4.00 equiv).
The resulting solution was stirred for 2 h at room temperature. The
resulting solution was extracted with 2.times.10 mL of ethyl
acetate and the organic layers were combined. The resulting mixture
was washed with 4.times.5 mL of water. The mixture was dried over
anhydrous sodium sulfate. The solids were filtered out. The
resulting mixture was concentrated under vacuum. This resulted in
110 mg (47%) of
(2S,4R)-1-[(2S)-3,3-dimethyl-2-(3-[2-[(5-[[(4-methylbenzene)-sulfonyl]oxy-
]pentyl)oxy]ethoxy]propanamido)butanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thi-
azol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide as colorless oil.
LC-MS m/z: (ES+) [M+H]+=787; Retention time: 1.03 min.
11. Step--Synthesis of
(2S,4R)-1-[(2S)-2-[3-(2-[[5-([4-[(3-Chloro-4-fluorophenyl)amino]-7-methox-
yquinazolin-6-yl]oxy)pentyl]oxy]ethoxy)propanamido]-3,3-dimethylbutanoyl]--
4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-car-
boxamide
##STR00573##
[1472] Into a 50 mL round-bottom flask, was placed
4-[(3-chloro-4-fluorophenyl)amino]-7-methoxyquinazolin-6-ol (45 mg,
0.14 mmol, 1.00 equiv),
(2S,4R)-1-[(2S)-3,3-dimethyl-2-(3-[2-[(5-[[(4-methylbenzene)sulfonyl]oxy]-
-pentyl)oxy]ethoxy]propanamido)butanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thi-
azol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (110 mg, 0.14
mmol, 1.00 equiv), potassium carbonate (58 mg, 0.42 mmol, 3.00
equiv), N,N-dimethylformamide (2 mL). The resulting solution was
stirred for 4 h at 80.degree. C. The solids were filtered out. The
crude product was purified by Prep-HPLC with the following
conditions: XBridge Prep C18 OBD Column, 19.times.100 mm, 5 micron;
mobile phase, water with 0.1% TFA and MeCN (25.0% MeCN up to 45.0%
in 10 min); Detector, UV 254 nm. HPLC purification resulted in 16.3
mg (12%) of
(2S,4R)-1-[(2S)-2-[3-(2-[[5-([4-[(3-chloro-4-fluorophenyl)amino]-7-methox-
yquinazolin-6-yl]oxy)pentyl]
oxy]ethoxy)propanamido]-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1-
,3-thiazol-5-yl) phenyl]methyl]pyrrolidine-2-carboxamide as a white
solid. LC-MS m/z: (ES+) [M+H]+=934; Retention time: 1.97 min; 1H
NMR (300 MHz, CD3OD, 25.degree. C.): 8.89 (s, 1H), 8.45 (s, 1H),
7.95-8.05 (d, 1H), 7.65-7.74 (m, 2H), 7.36-7.49 (m, 3H), 7.17-7.31
(m, 2H), 4.68-4.34 (m, 5H), 4.15-4.23 (m, 2H), 4.02 (s, 3H),
3.91-3.72 (m, 4H), 3.62 (s, 4H), 3.52-3.54 (m, 2H), 2.58-2.45 (m,
4H), 2.25-1.52 (m, 8H), 1.05 (s, 9H).
Synthesis of Example 41
(2S,4R)-1-((S,E)-2-(tert-butyl)-16-((4-((4-chloro-3-fluorophenyl)amino)-7--
methoxyquinazolin-6-yl)amino)-12-methyl-4,16-dioxo-6,9-dioxa-3,12-diazahex-
adec-14-enoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-c-
arboxamide
##STR00574##
##STR00575##
[1473] 1. Step--Synthesis of (E)-4-bromobut-2-enoic acid
##STR00576##
[1475] To a solution of (E)-but-2-enoic acid (20 g, 0.23 mol) in
CCl.sub.4 were added NBS (43 g, 0.24 mol) and benzoyl peroxide (5.6
g, 0.023 mol) under N.sub.2. Then the mixture was stirred at
80.degree. C. for 3 h. TLC analysis showed the complete consumption
of (E)-but-2-enoic acid. It was cooled to rt. The mixture was
filtered through Celite, and the filtrate was concentrated under
vacuum. The residue was purified via column (PE:EA=5:1) to afford
the desired product (E)-4-bromobut-2-enoic acid (10 g, 26% yield)
as yellow white solid. .sup.1H-NMR (400 MHz, CDCl.sub.3): .delta.
7.01-7.09 (m, 1H), 5.98 (d, J=15.2 Hz, 1H), 3.96 (d, J=7.2 Hz,
2H).
2. Step--Synthesis of (E)-4-Bromobut-2-enoyl chloride
##STR00577##
[1477] To a solution of compound (E)-4-bromobut-2-enoic acid (5.2
g, 0.03 mol) in DCM was added SOCl.sub.2 (10 mL, 0.16 mol) dropwise
in an ice-bath. Then the mixture was stirred at rt overnight. The
solvent was removed under vacuum to afford the desired product
(E)-4-Bromobut-2-enoyl chloride as yellow oil, which was used into
next reaction without further purification.
3. Step--Synthesis of
N-(3-Chloro-4-fluorophenyl)-7-methoxy-6-nitroquinazolin-4-amine
##STR00578##
[1479] To a solution of
N-(3-chloro-4-fluorophenyl)-7-fluoro-6-nitroquinazolin-4-amine (15
g, 44.56 mmol) in MeOH 150 mL, was added 50% KOH (5 g, 89 mmol) at
rt. The reaction mixture was stirred at 70.degree. C. for 2 h. Then
it was cooled to rt and extracted with ethyl acetate. The combined
organic layers were washed with water, and dried over
Na.sub.2SO.sub.4. The organic phase was concentrated under vacuum
to afford
N-(3-Chloro-4-fluorophenyl)-7-methoxy-6-nitroquinazolin-4-amine (20
g, 96.7% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 10.16
(s, 1H), 9.21 (s, 1H), 8.67 (s, 1H), 8.15 (dd, J=2.4, 6.8 Hz, 1H),
7.79-7.81 (m, 1H), 7.48 (t, J=7.2 Hz, 2H), 4.07 (s, 3H).
4. Step--Synthesis of
N.sup.4-(3-Chloro-4-fluorophenyl)-7-methoxyquinazoline-4,6-diamine
##STR00579##
[1481] To a solution of
N-(3-Chloro-4-fluorophenyl)-7-methoxy-6-nitroquinazolin-4-amine (10
g, 28.6 mmol) in a mixture of ethanol (200 mL), THF (100 mL),
H.sub.2O (50 mL), and saturated NH.sub.4Cl solution (50 mL) was
added iron powder (6.5 g, 116 mmol) at rt. Then the mixture was
heated to 80.degree. C. for 3 h. The mixture was filtered through
Celite, and the cake was washed with ethanol. Water (100 mL) was
added to the filtrate, and the yellow white solid was formed. The
solid was filtered and dried to obtain the desired compound
N.sup.4-(3-chloro-4-fluorophenyl)-7-methoxyquinazoline-4,6-diami-
ne (8 g, 88% yield) as yellow solid. .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 9.41 (s, 1H), 8.38 (s, 1H), 8.20-8.17 (m,
1H), 7.82-7.80 (m, 1H), 7.41-7.37 (m, 2H), 5.36 (s, 2H), 3.97 (s,
3H).
5. Step--Synthesis of
(E)-4-Bromo-N-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yl)-
but-2-enamide
##STR00580##
[1483] To a solution of
N.sup.4-(3-chloro-4-fluorophenyl)-7-methoxyquinazoline-4,6-diamine
(10 g, 31.4 mmol) in THF were added (E)-4-Bromobut-2-enoyl chloride
(8.6 g, 47.1 mmol) and TEA (8 g, 78.4 mmol) at 0.degree. C.
subsequently. Then the mixture was stirred at room temperature for
2 h. The reaction mixture was quenched with water. The mixture was
extracted with EA. The combined organic layers were washed
NaHCO.sub.3solution and brine. The organic phase was dried over
Na.sub.2SO.sub.4, and concentrated to afford the desired compound
(E)-4-Bromo-N-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yl)-
but-2-enamide (7.6 g, 52% yield) as brown solid. .sup.1H-NMR (400
MHz, CDCl.sub.3): .delta. 8.95 (s, 1H), 8.65 (s, 1H), 8.25 (s, 1H),
7.93-7.91 (m, 1H), 7.72 (s, 1H), 7.54-7.51 (m, 1H), 7.14-7.10 (m,
1H), 6.56-6.48 (m, 2H), 4.06 (s, 3H), 3.46 (d, J=6.8 Hz, 2H). LCMS:
465 [M+H]; t.sub.R=1.38
##STR00581##
Step 6--Synthesis of tert-butyl
2-(2-(2-(tosyloxy)ethoxy)ethoxy)acetate
##STR00582##
[1485] To a stirred solution of tert-butyl
2-(2-(2-hydroxyethoxy)ethoxy)acetate (2.0 g, 9.08 mmol),
triethylamine (3.2 g, 31.78 mmol), and 4-dimethylaminopyridine (111
mg, 0.91 mmol) in anhydrous dichloromethane (20 ml) was added a
solution of 4-toluenesulfonyl chloride (1.9 g, 3.12 mmol) in
anhydrous dichloromethane (10 ml) dropwise at 0.degree. C. The
resulting mixture was then allowed to warm up to room temperature
and stirred at room temperature for 10 hours. TLC showed formation
of desired product. The reaction mixture was diluted with
dichloromethane (250 ml), washed with water (50 ml.times.3) and
brine (50 ml), dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to give a crude residue which
was purified by silica gel flash column chromatography (eluted with
20-30% ethyl acetate in hexane) to afford
tert-butyl2-(2-(2-(tosyloxy)ethoxy)ethoxy)acetate (1.7 g, yield
50%) as a colorless oil. LC_MS: (ES.sup.+): m/z 397.10
[M+Na.sup.+]. t.sub.R=2.799 min.
Step 7--Synthesis of
(E)-tert-butyl2-(2-(2-((4-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquin-
azolin-6-ylamino)-4-oxobut-2-enyl)(methyl)amino)ethoxy)ethoxy)acetate
##STR00583##
[1487] A mixture of
tert-butyl2-(2-(2-(tosyloxy)ethoxy)ethoxy)acetate (250 mg, 0.67
mmol) in methylamine methanol solution (30%, 5 ml) was stirred at
room temperature for 5 hours. TLC showed formation of desired
product. The volatiles were removed under reduced pressure; the
residue was partitioned between dichloromethane (120 ml) and water
(30 ml). The organic layer was collected, washed with brine (30
ml), dried over anhydrous sodium sulfate, and concentrated under
reduced pressure. The residue was re-dissolved in anhydrous
N-methyl-2-pyrrolidone (3 ml), followed by sequentially addition of
N,N-diisopropylethylamine (30 mg, 0.23 mmol) and
(E)-4-bromo-N-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yl)-
but-2-en amide (100 mg, 0.21 mmol) at room temperature. The
resulting mixture was stirred at room temperature for 5 hours. TLC
showed formation of desired product. The reaction mixture was
partitioned between ethyl acetate (150 ml) and water (40 ml). The
organic layer was collected, washed with brine (30 ml), dried over
anhydrous sodium sulfate, and concentrated under reduced pressure
to give a crude residue which was purified by pre-TLC to afford
(E)-tert-butyl2-(2-(2-((4-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquin-
azolin-6-ylamino)-4-oxobut-2-enyl)(methyl)amino)ethoxy)ethoxy)acetate
(60 mg, yield 45%) as yellow oil. LC_MS: (ES.sup.+): m/z 618.30
[M+H.sup.+]. t.sub.R=1.696 min.
Step 8--Synthesis of
(2S,4R)-1-((S,E)-17-(4-(4-chloro-3-fluorophenylamino)-7-methoxyquinazolin-
-6-ylamino)-2,2,13-trimethyl-5,17-dioxo-7,10-dioxa-4,13-diazaheptadec-15-e-
necarbonyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carb-
oxamide
##STR00584##
[1489] A mixture of
(E)-tert-butyl2-(2-(2-((4-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquin-
azolin-6-ylamino)-4-oxobut-2-enyl)(methyl)amino)ethoxy)ethoxy)acetate
(60 mg, 0.097 mmol) in 2,2,2-trifluoroacetic acid (1 ml) and
anhydrous dichloromethane (1 ml) was stirred at room temperature
for 2 hours. TLC showed formation of desired product. The volatiles
were evaporated under reduced pressure; the residue was
re-dissolved in anhydrous N,N-dimethylformamide (2 ml), followed by
sequential addition of
(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthia-
zol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloric acid salt
(46 mg, 0.098 mmol), N,N-diisopropylethylamine (51 mg, 0.388 mmol),
and HATU
(2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate)(74 mg, 0.194 mmol) at 0.degree. C. The
resulting mixture was allowed to warm up to room temperature and
stirred at room temperature for 20 min. TLC showed formation of
desired product. The reaction mixture was partitioned between ethyl
acetate (150 ml) and water (30 ml). The organic layer was
collected, washed with brine (20 ml), dried over anhydrous sodium
sulfate, and concentrated under reduced pressure to give a crude
residue which was purified by pre-TLC to afford
(2S,4R)-1-((S,E)-17-(4-(4-chloro-3-fluorophenylamino)-7-methoxyquinazolin-
-6-ylamino)-2,2,13-trimethyl-5,17-dioxo-7,10-dioxa-4,13-diazaheptadec-15-e-
necarbonyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carb-
oxamide (26.9 mg, yield 28%) as a light yellow solid. LC_MS:
(ES.sup.+): m/z 974.20 [M+H.sup.+]. t.sub.R=1.630 min. .sup.1H NMR
(400 MHz, CD.sub.3OD): .delta.1.04, 1.06 (two singles, 9H),
2.06-2.11 (m, 1H), 2.25-2.30 (m, 1H), 2.44 (s, 3H), 2.83, 2.85 (two
singles, 3H), 3.77-4.16 (m, 15H), 4.42-4.63 (m, 4H), 4.76-4.81 (m,
1H), 6.76 (d, J=15.2 Hz, 1H), 7.04-7.08 (m, 1H), 7.24-7.28 (m, 2H),
7.36-7.40 (m, 4H), 7.67-7.68 (m, 1H), 7.99 (br, 1H), 8.37 (br, 1H),
8.49 (s, 1H), 8.85 (s, 1H), 8.93 (s, 1H).
Synthesis of Example 59
(2S,4R)-1-((S)-2-(6-(2-((4-((3-chloro-4-fluorophenyl)amino)-6-((E)-4-(dime-
thylamino)but-2-enamido)quinazolin-7-yl)oxy)ethoxy)hexanamido)-3,3-dimethy-
lbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carbo-
xamide
##STR00585##
##STR00586## ##STR00587##
[1490] Experimental Details
1. Step--Synthesis of
4-[(3-chloro-4-fluorophenyl)amino]-6-nitroquinazolin-7-ol
##STR00588##
[1492] Into a 250-mL round-bottom flask, was placed a solution of
N-(3-chloro-4-fluorophenyl)-7-fluoro-6-nitroquinazolin-4-amine (5.0
g, 14.85 mmol, 1.00 equiv) in dixoane/50% sodium hydroxide (aq)
(80/15 mL). The resulting solution was heated to reflux for 2 hr.
The reaction mixture was cooled. The resulting solution was diluted
with water (1000 mL). The pH value of the solution was adjusted to
3 with hydrogen chloride(c). The resulting solution was extracted
with ethyl acetate (500 mL.times.4) and the organic layers
combined. The resulting mixture was washed with brine (500
mL.times.1). The mixture was dried over anhydrous sodium sulfate
and concentrated under vacuum. The residue was applied onto a
silica gel column with ethyl acetate/petroleum ether (1:2). This
resulted in 1.92 g (39%) of
4-[(3-chloro-4-fluorophenyl)amino]-6-nitroquinazolin-7-ol as a
orange solid. .sup.1H NMR (300 MHz, DMSO) .delta. 11.94-11.81 (b,
1H), 10.17 (s, 1H), 9.22 (s, 1H), 8.60 (s, 1H), 8.16-8.15 (d, J=4.5
Hz, 1H), 7.81-7.78 (d, J=8.1 Hz, 1H), 7.53-7.44 (m, 2H), 7.26 (s,
1H); LC-MS (ES.sup.+): m/z 335.05 [MH.sup.+], t.sub.R0.95 min (1.9
minute run).
2. Step--Synthesis of
6-amino-4-[(3-chloro-4-fluorophenyl)amino]quinazolin-7-ol
##STR00589##
[1494] Into a 250-mL round-bottom flask, was placed a solution of
4-[(3-chloro-4-fluorophenyl)amino]-6-nitroquinazolin-7-ol (4.0 g,
11.95 mmol, 1.00 equiv) in tetrahydrofuran (100 mL), Raney Ni (2.0
g) was added in the solution under nitrogen atmosphere. Nitrogen
was removed under vacuum, hydrogen was introduced into under
hydrogen atmosphere. The resulting solution was stirred for 2 h at
room temperature. The solids were filtered out. The resulting
mixture was concentrated under vacuum. The residue was applied onto
a silica gel column with dichloromethane/methanol (10:1). This
resulted in 2.3 g (63%) of
6-amino-4-[(3-chloro-4-fluorophenyl)amino]quinazolin-7-ol as a blue
green solid. .sup.1H NMR (300 MHz, DMSO) .delta. 10.80-10.50 (b,
1H), 9.33 (s, 1H), 8.37 (s, 1H), 8.19-8.18 (d, J=4.5 Hz, 1H),
7.81-7.78 (d, J=8.1 Hz, 1H), 7.41-7.35 (m, 2H), 7.00 (s, 1H),
5.30-5.22 (b, 2H); LC-MS (ES.sup.+): m/z 305.20 [MH.sup.+],
t.sub.R0.85 min (1.9 minute run).
3. Step--Synthesis of
(2E)-N-[4-[(3-chloro-4-fluorophenyl)amino]-7-hydroxyquinazolin-6-yl]-4-(d-
imethylamino)but-2-enamide
##STR00590##
[1496] Into a 100-mL round-bottom flask purged and maintained with
an inert atmosphere of nitrogen, was placed a solution of
(2E)-4-(dimethylamino)but-2-enoic acid hydrogen chloride (1.27 g,
9.83 mmol, 2.00 equiv) in acetonitrile (25 mL).
N,N-dimethylformamide (3 drops) (cat) and oxalic dichloride (10 mL)
were added dropwise at room temperature. The resulting solution was
stirred for 30 min at 55.degree. C. The solution was concentrated
under vacuum. This black oil was dissolved in
1-Methyl-2-pyrrolidinone (10 mL), and was dropwised into another a
100-mL round-bottom flask purged and maintained with an inert
atmosphere of nitrogen, that was loaded with a solution of
6-amino-4-[(3-chloro-4-fluorophenyl)amino]quinazolin-7-ol (1.5 g,
4.92 mmol, 1.00 equiv) in 1-Methyl-2-pyrrolidinone (40 mL) and
sodium bicarbonate (4.0 g, 37.74 mmol, 7.00 equiv) at 0.degree. C.
The resulting solution was allowed to react, with stirring, for 2 h
at room temperature. The reaction was then quenched by the addition
of 30 mL of water. The solids were removed by filtration. The
resulting mixture was concentrated under vacuum. The residue was
applied onto C18 column (330 g) with water/methanol (10%-70%) in 90
mins. This resulted in 2.03 g (80% purity) of
(2E)-N-[4-[(3-chloro-4-fluorophenyl)amino]-7-hydroxyquinazolin-6-yl]-4-(d-
imethylamino)but-2-enamide as a brown solid. LC-MS (ES.sup.+): m/z
416.05[MH.sup.+], t.sub.R=1.05 min (2.6 minute run).
4. Step--Synthesis of tert-butyl
6-(2-[[(4-methylbenzene)sulfonyl]oxy]ethoxy)hexanoate
##STR00591##
[1498] Into a 100-mL round-bottom flask, was placed tert-butyl
6-(2-hydroxyethoxy)hexanoate (500.0 mg, 2.15 mmol, 1.00 equiv),
triethylamine (653.0 mg, 6.45 mmol, 3.00 equiv),
4-methylbenzene-1-sulfonyl chloride (614.0 mg, 3.22 mmol, 1.50
equiv), 4-dimethylaminopyridine (26 mg, 0.21 mmol, 0.10 equiv),
dichloromethane (25 mL). The resulting solution was stirred for 12
h at room temperature. The resulting mixture was concentrated under
vacuum. The residue was applied onto a silica gel column with ethyl
acetate/petroleum ether (1:2). This resulted in 650.0 mg (78%) of
tert-butyl 6-(2-[[(4-methylbenzene)sulfonyl]oxy]ethoxy)hexanoate as
colorless oil.
5. Step--Synthesis of tert-butyl
6-[2-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-2--
enamido]quinazolin-7-yl]oxy)ethoxy]hexanoate
##STR00592##
[1500] Into a 50-mL round-bottom flask, was placed a solution of
(2E)-N-[4-[(3-chloro-4-fluorophenyl)amino]-7-hydroxyquinazolin-6-yl]-4-(d-
imethylamino)but-2-enamide (160.0 mg, 0.38 mmol, 1.00 equiv) in
N,N-dimethylformamide (15.0 mL), Cs.sub.2CO.sub.3 (251.0 mg, 0.77
mmol, 2.00 equiv), tert-butyl
6-(2-[(4-methylbenzene)sulfonyl]oxyethoxy)hexanoate (148.0 mg, 0.38
mmol, 1.00 equiv). The resulting solution was stirred for 3 h at
80.degree. C. The reaction was then quenched by the addition of
water (10 mL). The resulting solution was extracted with ethyl
acetate (20 mL.times.2) and the organic layers combined. The
resulting mixture was washed with brine (30 mL.times.1). The
mixture was dried over anhydrous sodium sulfate and concentrated
under vacuum. The residue was applied onto a silica gel column with
ethyl acetate/petroleum ether (1:2). The collected fractions were
combined and concentrated under vacuum. This resulted in 130.0 mg
(54%) of tert-butyl
6-[2-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-2--
enamido]quinazolin-7-yl]oxy)ethoxy]hexanoate as yellow oil. LC-MS
(ES.sup.+): m/z 630.35 [MH.sup.+], t.sub.R=1.33 min 1.9 minute
run).
6. Step--Synthesis of
6-[2-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-2--
enamido]quinazolin-7-yl]oxy)ethoxy]hexanoic acid
##STR00593##
[1502] Into a 50-mL round-bottom flask, was placed a solution of
tert-butyl
6-[2-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-2--
enamido]quinazolin-7-yl]oxy)ethoxy]hexanoate (130.0 mg, 0.21 mmol,
1.00 equiv) in dichloromethane/trifluoroacetic acid (10/3 mL). The
resulting solution was stirred for 2 h at room temperature. The
resulting mixture was concentrated under vacuum. This resulted in
100.0 mg (84%) of
6-[2-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-2--
enamido]quinazolin-7-yl]oxy)ethoxy]hexanoic acid as yellow oil.
LC-MS (ES.sup.+): m/z 574.15[MH.sup.+], t.sub.R=0.53 min (1.9
minute run).
7. Step--Synthesis of
(2S,4R)-1-[(2S)-2-[6-[2-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(d-
imethylamino)but-2-enamido]quinazolin-7-yl]oxy)ethoxy]hexanamido]-3,3-dime-
thylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyr-
rolidine-2-carboxamide
##STR00594##
[1504] Into a 50-mL round-bottom flask, was placed a solution of
6-[2-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-2--
enamido]quinazolin-7-yl]oxy)ethoxy]hexanoic acid (100.0 mg, 0.17
mmol, 1.00 equiv) in N,N-dimethylformamide (10.0 mL),
N-ethyl-N-isopropylpropan-2-amine (90.0 mg, 0.70 mmol, 4.00 equiv).
This was followed by the addition of
o-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (80.0 mg, 0.21 mmol, 1.20 equiv). It was
stirred for 5 min at room temperature. To this was added
(2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[4-(4-methyl-1,-
3-thiazol-5-yl)phenyl]methylpyrrolidine-2-carboxamide hydrochloride
(98.0 mg, 0.21 mmol, 1.20 equiv). The resulting solution was
stirred for 2 h at room temperature. The reaction was then quenched
by the addition of water (20 mL). The resulting solution was
extracted with ethyl acetate (20 mL.times.3) and the organic layers
combined. The resulting mixture was washed with brine (10
mL.times.1). The solid was dried in an oven under reduced pressure.
The crude product was purified by Prep-HPLC with the following
conditions: Column: X Bridge C18, 19*150 mm, 5 um; Mobile Phase A:
Water/10 mmol/L Ammonium bicarbonate Mobile Phase B: acetonitrile;
Flow rate: 20 mL/min; Gradient: 30% B to 65% B in 8 min; 254 nm.
This resulted in 12.5 mg (7%) of
(2S,4R)-1-[(2S)-2-[6-[2-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(d-
imethylamino)but-2-enamido]quinazolin-7-yl]oxy)ethoxy]hexanamido]-3,3-dime-
thylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyr-
rolidine-2-carboxamide as a yellow solid. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.92 (s, 1H), 8.83 (s, 1H), 8.45 (s, 1H),
8.01-7.99 (d, J=8.4 Hz, 1H), 7.70-7.60 (m, 1H), 7.43-7.36 (m, 4H),
7.25-7.19 (m, 2H), 7.02-6.97 (m, 1H), 6.50-6.45 (d, J=15.3 Hz, 1H),
4.58 (s, 1H), 4.52-4.47 (m, 3H), 4.39-4.33 (m, 3H), 3.93-3.92 (m,
2H), 3.90-3.83 (m, 1H), 3.82-3.70 (m, 1H), 3.60-3.56 (m, 2H),
3.20-3.18 (m, 2H), 2.42 (s, 3H), 2.29 (s, 6H), 2.29-2.12 (m, 3H),
2.10-2.00 (m, 1H), 1.65-1.60 (m, 4H), 1.38-1.29 (m, 2H), 0.97 (s,
9H); LC-MS (ES.sup.+): m/z 986.40 [MH.sup.+], t.sub.R=0.82 min (3.0
minute run). Chemical formula: C.sub.50H.sub.61ClFN.sub.9O.sub.7S
[985.41]
Synthesis of Example 66
(2S,4R)-1-((S)-2-(6-(4-(4-((4-((3-chloro-4-fluorophenyl)amino)-6-((E)-4-(d-
imethylamino)but-2-enamido)quinazolin-7-yl)oxy)butoxy)butoxy)hexanamido)-3-
,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidi-
ne-2-carboxamide
##STR00595##
[1506] Synthetic Scheme:
##STR00596##
1. Step--Synthesis of tert-Butyl
6-[4-[4-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-
-2-enamido]quinazolin-7-yl]oxy)butoxy]butoxy]hexanoate
##STR00597##
[1508] Into a 25 mL round-bottom flask, was placed a solution of
(2E)-N-[4-[(3-chloro-4-fluorophenyl)amino]-7-hydroxyquinazolin-6-yl]-4-(d-
imethylamino)but-2-enamide (200.0 mg, 0.48 mmol, 1.00 equiv) in
N,N-dimethylformamide (10 mL), cesium carbonate (314.0 mg, 0.96
mmol, 2.00 equiv), tert-butyl
6-(4-(4-(tosyloxy)butoxy)butoxy)hexanoate (281.0 mg, 1.20 equiv).
The resulting solution was stirred for 12 h at 80 DC in an oil
bath. The reaction was then quenched by the addition of water (20
mL). The resulting mixture was extracted with ethyl acetate (10
mL.times.3) and the organic layers were combined. The resulting
mixture was washed with brine (10 mL). The mixture was dried over
anhydrous sodium sulfate and concentrated under vacuum. The residue
was applied onto a silica gel column eluted with
dichloromethane/methanol (10:1). This resulted in 77.0 mg (22%) of
tert-butyl
6-[4-[4-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-
-2-enamido]quinazolin-7-yl]oxy)butoxy]butoxy]hexanoate as a brown
solid. LC-MS (ES.sub.+): m/z 730.31 [M+H].sub.+
2. Step--Synthesis of
6-[4-[4-([4-[(3-Chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-
-2-enamido]quinazolin-7-yl]oxy)butoxy]butoxy]hexanoic acid
##STR00598##
[1510] Into a 25 mL round-bottom flask, was placed a solution of
tert-butyl
6-[4-[4-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-
-2-enamido]quinazolin-7-yl]oxy)butoxy]butoxy]hexanoate (77.0 mg,
0.11 mmol, 1.00 equiv) in dichloromethane/trifluoroacetic acid
(10/2 mL). The resulting solution was stirred for 2 h at room
temperature. The resulting mixture was concentrated under vacuum.
This resulted in 71.0 mg (100%) of
6-[4-[4-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-
-2-enamido]quinazolin-7-yl]oxy)butoxy]butoxy]hexanoic acid as brown
oil. LC-MS (ES+): m/z 674.30 [M+H]+
3. Step--Synthesis of
(2S,4R)-1-[(2S)-2-(6-[4-[4-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-
-(dimethylamino)but-2-enamido]quinazolin-7-yl]oxy)butoxy]butoxy]hexanamido-
)-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]-
methyl]pyrrolidine-2-carboxamide
##STR00599##
[1512] Into a 25 mL round-bottom flask under ice bath, was placed a
solution of
6-[4-[4-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-(dimethylamino)but-
-2-enamido]quinazolin-7-yl]oxy)butoxy]butoxy]hexanoic acid (71.0
mg, 0.11 mmol, 1.00 equiv) in N,N-dimethylformamide (10 mL),
N-ethyl-N-isopropylpropan-2-amine (68.0 mg, 0.53 mmol, 5.00 equiv),
o-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (52.0 mg, 0.14 mmol, 1.30 equiv). The mixture
was stirred for 20 min. To this was added
(2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[4-(4-methyl-1,-
3-thiazol-5-yl)phenyl]methylpyrrolidine-2-carboxamide hydrochloride
(64.0 mg, 1.30 equiv) at 0.degree. C. The resulting solution was
stirred for 2 h at room temperature. The resulting solution was
extracted with ethyl acetate (10 mL.times.3) and the organic layers
were combined. After washing with brine (10 mL), the mixture was
dried over anhydrous sodium sulfate, filtered, and concentrated
under vacuum. The residue was purified by prep-HPLC under the
following condition: X Bridge RP18 column, 19.times.150 mm, 5
micron; Mobile Phase A: water/0.05% ammonium bicarbonate; Mobile
Phase B: acetonitrile; Flow rate: 20 mL/min; Gradient: 38% B to 52%
B in 15 min; Detector: 254 nm. This resulted in 27.0 mg (24%) of
(2S,4R)-1-[(2S)-2-(6-[4-[4-([4-[(3-chloro-4-fluorophenyl)amino]-6-[(2E)-4-
-(dimethylamino)but-2-enamido]quinazolin-7-yl]oxy)butoxy]butoxy]hexanamido-
)-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]-
methyl]pyrrolidine-2-carboxamide as a white solid. 1H NMR (300 MHz,
CD3OD): .delta. 8.88 (s, 1H), 8.49 (s, 1H), 8.04-8.02 (d, J=6.9 Hz,
1H), 7.75-7.65 (m, 1H), 7.47-7.41 (m, 4H), 7.38-7.26 (m, 2H),
7.05-7.00 (m, 1H), 6.55-6.50 (d, J=15.3 Hz, 1H), 4.63 (s, 1H),
4.56-4.49 (m, 3H), 4.37-4.30 (m, 3H), 3.92-3.78 (m, 2H), 3.57-3.53
(m, 2H), 3.47-3.46 (m, 2H), 3.46-3.40 (m, 4H), 3.24-3.21 (m, 2H),
2.47 (s, 3H), 2.32 (s, 6H), 2.30-2.23 (m, 3H), 2.08-2.01 (m, 3H),
1.84-1.82 (m, 2H), 1.61-1.52 (m, 8H), 1.38-1.25 (m, 2H), 1.00 (s,
9H); LC-MS (ES+): m/z 544.25 [(M+2H+)/2].
Synthesis of Example 70
2-(2,6-dioxopiperidin-3-yl)-5-(2-(2-(2-((1-isopropyl-6-((2-(4-methoxypiper-
idin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)etho-
xy)ethoxy)ethoxy)isoindoline-1,3-dione
##STR00600##
[1514] Synthetic Scheme:
##STR00601## ##STR00602##
1. Step--Synthesis of 1-Phenyl-2,5,8,11-tetraoxatridecan-13-ol
##STR00603##
[1516] To a solution of 3,6,9,12-tetraoxatetradecane-1,14-diol (45
g, 234 mmol) in DMF (100 mL), was added NaH (60%, 2.34 g, 58.5
mmol) at 0.degree. C. After stirring at RT 1 h, BnBr (10 g, 58.5
mmol) was added and the mixture was heated to 60.degree. C. for 3
h. Then the reaction mixture was quenched with water (100 mL) and
the resulting reaction mixture was extracted with EtOAc
(2.times.200 mL). The combined organic layers were washed with
brine (100 mL), dried over Na.sub.2SO.sub.4 and filtered. The
solvent was evaporated under reduced pressure. The residue was
purified by column chromatography (hexane:EtOAc1:3) to afford the
desired product 1-Phenyl-2,5,8,11-tetraoxatridecan-13-ol (15 g,
52.8 mmol, 90%). .sup.1H NMR (400 MHz, CDCl3): .delta. 7.27-7.34
(m, 5H), 4.57 (s, 2H), 3.59-3.73 (m, 16H). Chemical Formula:
C.sub.15H.sub.24O.sub.5; Molecular Weight: 284.35
2. Step--Synthesis of 1-Phenyl-2,5,8,11-tetraoxatridecan-13-oic
acid
##STR00604##
[1518] To a solution of 1-Phenyl-2,5,8,11-tetraoxatridecan-13-ol
(6.3 g, 22.1 mmol) in DCM (60 mL) and H.sub.2O (30 mL) were added
PhI(OAc)2 (21.3 g, 66.3 mmol and TEMPO (689 mg, 4.42 mmol)
subsequently at 0.degree. C. The mixture was stirred at room
temperature for 3 h. The reaction mixture was diluted with water
(200 mL) and extracted with DCM. The organic layer was washed with
brine, dried over MgSO.sub.4, and concentrated in vacuo. The
residue was purified by column chromatography (DCM:MeOH=20:1) to
afford the desired product
1-Phenyl-2,5,8,11-tetraoxatridecan-13-oic acid (4.0 g, 13.4 mmol,
60.6%). .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.27-7.35 (m,
5H), 4.57 (s, 2H), 4.13 (s, 2H), 3.63-3.75 (m, 12H). Chemical
Formula: C.sub.15H.sub.22O.sub.6; Molecular Weight: 298.34
3. Step--Synthesis of
N-(4-(Isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)pyridin-3-yl)-1-phenyl-2,5,8,11-tetraoxatridecan-13-amide
##STR00605##
[1520] To a solution of 1-Phenyl-2,5,8,11-tetraoxatridecan-13-oic
acid (1.05 g, 3.53 mmol),
N.sup.4-isopropyl-N.sup.2-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)pyri-
dine-2,4,5-triamine (1.05 g, 2.94 mmol, J. Med. Chem. 2015, 58,
8877-8895), Et.sub.3N (742 mg, 7.35 mmol) and HOBt (595 mg, 4.41
mmol) in DCM (60 mL) was added EDCI (842 mg, 4.41 mol) at RT. The
reaction mixture was stirred at rt for 2 h. Then the reaction
mixture was diluted with water (20 mL) and the resulting reaction
mixture was extracted with DCM (2.times.50 mL). The combined
organic layers were washed with brine (100 mL), dried over
Na.sub.2SO.sub.4 and filtered. The solvent was evaporated under
reduced pressure. The residue was purified by column chromatography
(DCM:MeOH=20:1) to afford the desired compound
N-(4-(Isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)pyridin-3-yl)-1-phenyl-2,5,8,11-tetraoxatridecan-13-amide (1.4 g,
2.19 mmol, 74.8%). LC-MS: (ES.sup.+): m/z 638.3 [M+H]. t.sub.R=3.46
min. Chemical Formula: C.sub.33H.sub.47N.sub.7O.sub.6; Molecular
Weight: 637.78
4. Step--Synthesis of
1-Isopropyl-N-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-2-(12-phenyl-2,-
5,8,11-tetraoxadodecyl)-1H-imidazo[4,5-c]pyridin-6-amine
##STR00606##
[1522] A solution of
N-(4-(Isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)pyridin-3-yl)-1-phenyl-2,5,8,11-tetraoxatridecan-13-amide (1.4 g,
2.19 mmol) in HOAc (5 mL) was irradiated with microwave at
150.degree. C. for 6 h. The solvent was removed in vacuo to afford
crude desired product
1-Isopropyl-N-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-2-(12-phenyl-2,-
5,8,11-tetraoxadodecyl)-1H-imidazo[4,5-c]pyridin-6-amine (1.4 g,
crude), which was used into next reaction without further
purification. LC-MS: (ES.sup.+): m/z 620.3 [M+H]. t.sub.R=3.24 min
Chemical Formula: C.sub.33H.sub.45N.sub.7O.sub.5; Molecular Weight:
619.77.
5. Step--Synthesis of
2-(2-(2-((1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethoxy)ethoxy)ethan-1-ol
##STR00607##
[1524] To a stirred solution of
1-Isopropyl-N-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-2-(12-phenyl-2,-
5,8,11-tetraoxadodecyl)-1H-imidazo[4,5-c]pyridin-6-amine (1.2 g,
crude) in methanol (50 mL) was added Pd(OH).sub.2/C (10%, 0.5 g)
and cat. conc. HCl (0.1 mL). The mixture was stirred for 2 h under
H.sub.2 1 atm. The mixture was filtered through a Celite pad, and
the filtrate was concentrated. The residue was dissolved with DCM,
and the mixture was washed with aq.NaHCO.sub.3, brine and dried
with Na.sub.2SO.sub.4. The solvent was removed under vacuum to
afford the desired compound
2-(2-(2-((1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethoxy)ethoxy)ethan-1-ol
(1.0 g, crude). LC-MS: (ES.sup.+): m/z 530.3 [M+H]. t.sub.R=2.59
min. Chemical Formula: C.sub.26H.sub.39N.sub.7O.sub.5; Molecular
Weight: 529.64
6. Step--Synthesis of Tert-butyl
5-amino-4-(5-(2-(2-(2-((1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimi-
din-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethoxy)ethoxy)ethoxy-
)-1,3-dioxoisoindolin-2-yl)-5-oxopentanoate
##STR00608##
[1526] To a solution of
2-(2-(2-((1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethoxy)ethoxy)ethan-1-ol
(265 mg, 0.50 mmol), PPh.sub.3 (655 mg, 2.5 mmol) and tert-butyl
5-amino-4-(5-hydroxy-1,3-dioxoisoindolin-2-yl)-5-oxopentanoate (175
mg, 0.50 mmol) in dry THF (10 mL) was added DIAD (505 mg, 2.5 mmol)
dropwise at 0.degree. C. under N.sub.2. The mixture was stirred at
room temperature for 1 h. Then the reaction mixture was diluted
with water (50 mL) and extracted with DCM. The organic phase was
washed with brine, dried over MgSO.sub.4, and concentrated. The
residue was purified by chromatography (silica gel, DCM: MeOH
(50:1, v:v)) to afford the desired compound Tert-butyl
5-amino-4-(5-(2-(2-(2-((1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimi-
din-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethoxy)ethoxy)ethoxy-
)-1,3-dioxoisoindolin-2-yl)-5-oxopentanoate (90 mg, crude).
Chemical Formula: C43H57N9O10; Molecular Weight: 859.98
7. Step--Synthesis of
2-(2,6-Dioxopiperidin-3-yl)-5-(2-(2-(2-((1-isopropyl-6-((2-(4-methoxypipe-
ridin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)eth-
oxy)ethoxy)ethoxy)isoindoline-1,3-dione
##STR00609##
[1528] To a solution of Tert-butyl
5-amino-4-(5-(2-(2-(2-((1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimi-
din-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethoxy)ethoxy)ethoxy-
)-1,3-dioxoisoindolin-2-yl)-5-oxopentanoate (80 mg, 0.093 mmol) in
CH.sub.3CN (5 mL) was added TsOH (62 mg, 0.36 mmol). The solution
was stirred at 80.degree. C. for 3 h. Then it was cooled to RT and
quenched with aq NaHCO.sub.3. The mixture was taken up with DCM.
The organic phase was washed with water (10 mL.times.2) and brine
(10 mL). The organic layer was dried over anhydrous sodium sulfate,
filtered and concentrated under vacuum. The residue was purified by
silica gel column chromatography (DCM/MeOH=20/1) to afford
2-(2,6-dioxopiperidin-3-yl)-5-(2-(2-(2-((1-isopropyl-6-((2-(4-methoxypipe-
ridin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)eth-
oxy)ethoxy)ethoxy)isoindoline-1,3-dione (12 mg, 0.015 mmol, 16%).
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 12.59 (s, 1H), 8.65-8.71
(m, 3H), 8.12 (s, 1H), 7.72 (d, J=8.0 Hz, 1H), 7.32 (s, 1H), 7.17
(d, J=8.0 Hz, 1H), 6.60 (s, 1H), 5.30-5.31 (m, 1H), 4.94-4.96 (m,
2H), 4.24 (s, 2H), 3.87-4.03 (m, 2H), 3.72-3.85 (m, 4H), 3.60-3.82
(m, 10H), 3.39 (s, 3H), 2.71-2.92 (m, 3H), 2.09-2.11 (m, 1H),
1.86-2.00 (m, 2H), 1.75-1.86 (m, 2H), 1.66-1.68 (m, 6H). LC-MS:
(ES.sup.+): m/z 786.2 [M+H]. t.sub.R=3.10 min Chemical Formula:
C.sub.39H.sub.47N.sub.9O.sub.9; Molecular Weight: 785.86
Synthesis of Example 71
2-(2,6-dioxopiperidin-3-yl)-5-(4-(2-((1-isopropyl-6-((2-(4-methoxypiperidi-
n-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)p-
iperazin-1-yl)isoindoline-1,3-dione
##STR00610##
##STR00611##
[1529] Experimental Section
[1530]
N.sup.4-Isopropyl-N.sup.2-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-y-
l)pyridine-2,4,5-triamine was synthesized as described in J. Med.
Chem. 2015, 58, 8877-8895.
##STR00612##
[1531] .sup.1H NMR (400 MHz, MeOD): .delta. 8.56 (s, 1H), 8.06 (d,
J=5.6 Hz, 1H), 7.36 (s, 1H), 6.50 (s, 1H), 6.13 (d, J=5.6 Hz, 1H),
4.08-4.13 (m, 2H), 3.80-3.83 (m, 1H), 3.41-3.53 (m, 3H), 3.40 (s,
3H), 1.94-1.99 (m, 2H), 1.55-1.60 (m, 2H), 1.34 (d, J=6.4 Hz, 6H).
Chemical Formula: C.sub.18H.sub.27N.sub.7O; Molecular Weight:
357.46 LC-MS: (ES.sup.+): m/z 358.2 [M+H.sup.+]. t.sub.R=2.61
min.
##STR00613##
1. Step--Synthesis of Benzyl
4-(2-(2-hydroxyethoxy)ethyl)piperazine-1-carboxylate
##STR00614##
[1533] To a solution of 2-(2-hydroxyethoxy)ethyl
4-methylbenzenesulfonate (2.7 g, 10.4 mmol) and benzyl
piperazine-1-carboxylate (2.3 g, 10.4 mmol) in DMF (10 mL) was
added K.sub.2CO.sub.3 (2.86 g, 20.8 mmol). The solution was stirred
at 80.degree. C. overnight. The mixture was extracted with ethyl
acetate (50 mL.times.2). The organic phase was washed with water
(10 mL) and brine (8 mL). The organic layer was dried
(Na.sub.2SO.sub.4), filtered and concentrated under reduced
pressure to afford the crude Benzyl
4-(2-(2-hydroxyethoxy)ethyl)piperazine-1-carboxylate (4.5 g), which
was used in the next reaction without further purification.
2. Step--Synthesis of
2-(2-(4-((Benzyloxy)carbonyl)piperazin-1-yl)ethoxy)acetic acid
##STR00615##
[1535] To a solution of Benzyl
4-(2-(2-hydroxyethoxy)ethyl)piperazine-1-carboxylate (3.5 g, 11.4
mmol) and (Diacetoxyiodo)benzene (11.0 g, 34.2 mmol) in DCM (70 mL)
and water (35 ml) was added TEMPO (350 mg, 2.2 mmol). The solution
was stirred at rt for 4 h. The mixture was quenched with saturated
Na.sub.2S.sub.2O.sub.3 and stirred for 20 min. The mixture was
extracted with DCM (50 mL.times.2). The combined organic phases
were washed with water (10 mL) and brine (8 mL). The organic phase
was dried (Na.sub.2SO.sub.4), filtered and concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to afford
2-(2-(4-((Benzyloxy)carbonyl)piperazin-1-yl)ethoxy)acetic acid (300
mg, 0.93 mmol, 8.2% yield).
3. Step--Synthesis of Benzyl
4-(2-(2-((4-(isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-y-
l)amino)pyridin-3-yl)amino)-2-oxoethoxy)ethyl)piperazine-1-carboxylate
##STR00616##
[1537] To a solution of
2-(2-(4-((Benzyloxy)carbonyl)piperazin-1-yl)ethoxy)acetic acid (300
mg, 0.93 mmol) and
N.sup.4-isopropyl-N.sup.2-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)pyri-
dine-2,4,5-triamine (365 mg, 1.02 mmol) [J. Med. Chem. 2015, 58,
8877-8895] in DCM (15 mL) were added EDCI (231 mg, 1.2 mmol), HOBt
(162 mg, 1.2 mmol) and Et.sub.3N (151 mg, 1.5 mmol). The solution
was stirred at RT overnight and then diluted with DCM (50 mL). The
mixture was washed with water (10 ml) and brine (8 ml). The organic
layer was dried (Na.sub.2SO.sub.4), filtered and concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to afford Benzyl
4-(2-(2-((4-(isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-y-
l)amino)pyridin-3-yl)amino)-2-oxoethoxy)ethyl)piperazine-1-carboxylate
(380 mg, 0.57 mmol, 61.3% yield). Chemical Formula:
C.sub.34H.sub.47N.sub.9O.sub.5; Molecular Weight: 661.81 LC-MS:
(ES.sup.+): m/z 662.3 [M+H.sup.+]. t.sub.R=2.74 min
4. Step--Synthesis of Benzyl
4-(2-((1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1-
H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)piperazine-1-carboxylate
##STR00617##
[1539] A solution of Benzyl
4-(2-(2-((4-(isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin
-4-yl)amino)pyridin-3-yl)amino)-2-oxoethoxy)ethyl)piperazine-1-carboxylat-
e (380 mg, 0.57 mmol) in CH.sub.3COOH (8 mL) was irradiated by
microwave at 150.degree. C. for 6 h under nitrogen atmosphere. The
mixture was concentrated under reduced pressure to afford the crude
Benzyl
4-(2-((1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1-
H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)piperazine-1-carboxylate
(400 mg), which was used into the next reaction without further
purification. Chemical Formula: C.sub.34H.sub.45N.sub.9O.sub.4;
Molecular Weight: 643.79 LC-MS: (ES.sup.+): m/z 645.3 [M+H.sup.+].
t.sub.R=2.75 min.
5. Step--Synthesis of
1-Isopropyl-N-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-2-((2-(piperazi-
n-1-yl)ethoxy)methyl)-1H-imidazo[4,5-c]pyridin-6-amine
##STR00618##
[1541] To a solution of crude Benzyl
4-(2-((1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1-
H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)piperazine-1-carboxylate
(150 mg, crude) in methanol (12 mL) were added Pd(OH).sub.2 (10%,
70 mg) and conc. HCl (0.01 mL). The mixture was stirred for 2 h
under H.sub.2 atmosphere. The mixture was filtered through Celite,
and the filtrate was concentrated under reduced pressure to afford
crude
1-Isopropyl-N-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-2-((2-(piperazi-
n-1-yl)ethoxy)methyl)-1H-imidazo[4,5-c]pyridin-6-amine (120 mg),
which was used into the next reaction without further purification.
LC-MS: (ES.sup.+): m/z 510.3 [M+H].sup.+. t.sub.R=2.31 min.
Chemical Formula: C.sub.26H.sub.39N.sub.9O.sub.2; Molecular Weight:
509.66
6. Step--Synthesis of
2-(2,6-Dioxopiperidin-3-yl)-5-(4-(2-((1-isopropyl-6-((2-(4-methoxypiperid-
in-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)-
piperazin-1-yl)isoindoline-1,3-dione
##STR00619##
[1543] To a solution of
1-Isopropyl-N-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-2-((2-(piperazi-
n-1-yl)ethoxy)methyl)-1H-imidazo[4,5-c]pyridin-6-amine (110 mg,
0.21 mmol) and
2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (80 mg,
0.65 mmol) in NMP (8 mL) was added DIPEA (140 mg, 0.11 mmol). The
solution was irradiated by microwave at 150.degree. C. for 15 min
under nitrogen atmosphere. The mixture was concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to afford the crude desired product, and it was
further purified by prep-TLC to afford
2-(2,6-Dioxopiperidin-3-yl)-5-(4-(2-((1-isopropyl-6-((2-(4-methoxy-
piperidin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy-
)ethyl)piperazin-1-yl)isoindoline-1,3-dione (20 mg, 0.026 mmol,
12.4% yield). .sup.1H NMR (400 MHz, DMSO): .delta. 11.06 (s, 1H),
9.77 (s, 1H), 8.64 (s, 1H), 8.45 (s, 1H), 7.97 (d, J=6.4 Hz, 1H),
7.67 (d, J=8.4 Hz, 1H), 7.33 (s, 1H), 7.22 (d, J=8.4 Hz, 1H), 6.42
(d, J=6.0 Hz, 1H), 5.05-5.09 (m, 1H), 4.90-4.93 (m, 1H), 4.79 (s,
2H), 4.21-4.24 (m, 1H), 3.65 (s, 2H), 3.30-3.50 (m, 11H), 2.82-2.92
(m, 1H), 2.51-2.55 (m, 7H), 2.00-2.09 (m, 1H), 1.85-1.95 (m, 2H),
1.60 (d, J=6.8 Hz, 6H), 1.44-1.46 (m, 2H). Chemical Formula:
C.sub.39H.sub.47N.sub.11O.sub.6; Molecular Weight: 765.88 LC-MS:
(ES.sup.+): m/z 766.3 [M+H].sup.+. t.sub.R=2.67 min
Synthesis of Example 76
2-(2,6-dioxopiperidin-3-yl)-5-(2-(4-(2-((1-isopropyl-6-((2-(4-methoxypiper-
idin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethy-
l)piperazin-1-yl)ethoxy)isoindoline-1,3-dione
##STR00620##
[1545] Synthesis Scheme
##STR00621## ##STR00622##
Experimental Section
1. Step--Synthesis of Tert-butyl 2-(2-(benzyloxy)ethoxy)acetate
##STR00623##
[1547] To a solution of 2-(benzyloxy)ethanol (10.0 g, 0.07 mol) in
DCM (150 mL) were added tert-butyl 2-bromoacetate (51.0 g, 0.26
mol), TBACl (18.4 g, 0.07 mol) and 37% NaOH (15 mL) subsequently.
The resulting solution was stirred at r.t. overnight. After the
reaction was quenched with water (200 mL), the mixture was
extracted with DCM (2.times.200 mL). The combined organic layers
were washed with brine (100 mL), dried over Na.sub.2SO.sub.4 and
filtered. The solvent was removed under reduced pressure. The
residue was purified by column chromatography (DCM:MeOH=20:1) to
afford the desired product tert-butyl
2-(2-(benzyloxy)ethoxy)acetate (1.5 g, 5.60 mmol, 9%) as colorless
oil.
[1548] Chemical Formula: C.sub.15H.sub.22O.sub.4; Molecular Weight:
266.33
2. Step--Synthesis of Tert-butyl 2-(2-hydroxyethoxy)acetate
##STR00624##
[1550] To a solution of product tert-butyl
2-(2-(benzyloxy)ethoxy)acetate (1.50 g, 5.60 mmol) in MeOH (10 mL)
was added Pd(OH).sub.2/C (20%, 0.50 g). The mixture was stirred at
room temperature for 4 h under H.sub.2 at 1 atm. The reaction
mixture was filtered and concentrated in vacuo to afford the
desired product Tert-butyl 2-(2-hydroxyethoxy)acetate (1.65 g,
crude) as colorless oil. Chemical Formula: C.sub.8H.sub.16O.sub.4;
Molecular Weight: 176.21
3. Step Synthesis of Tert-butyl
2-(2-((methylsulfonyl)oxy)ethoxy)acetate
##STR00625##
[1552] To a solution of Tert-butyl 2-(2-hydroxyethoxy)acetate (1 g,
5.68 mmol) in DCM and Et.sub.3N (1.15 g, 11.36 mmol) at 0.degree.
C., MsCl (0.98 g, 8.52 mmol) was added. The reaction mixture was
stirred at r.t for 1 h. Then the reaction mixture was diluted with
20 mL water and the resulting reaction mixture was extracted with
DCM (2.times.50 mL). The combined organic layers were washed with
brine (1.times.100 mL), dried over Na.sub.2SO.sub.4 and filtered.
The solvent was removed under reduced pressure to afford the
desired product Tert-butyl 2-(2-((methylsulfonyl)oxy)ethoxy)acetate
(1.46 g, 5.75 mmol, crude) as colorless oil.
[1553] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 4.44 (t, J=4.4
Hz, 2H), 4.05 (s, 2H), 3.85 (t, J=4.4 Hz, 2H), 3.12 (s, 3H), 1.50
(s, 9H). Chemical Formula: C.sub.9H.sub.18O.sub.6S; Molecular
Weight: 254.30
4. Step--Synthesis of Tert-butyl
2-(2-(4-(2-(benzyloxy)ethyl)piperazin-1-yl)ethoxy)acetate
##STR00626##
[1555] A mixture of Tert-butyl
2-(2-((methylsulfonyl)oxy)ethoxy)acetate (1.46 g, 5.68 mmol),
1-(2-(benzyloxy)ethyl)piperazine (1.62 g, 7.38 mmol) and
K.sub.2CO.sub.3 (3.14 g, 22.70 mmol) in DMF (10 mL) was heated to
80.degree. C. overnight. After cooling to rt, the reaction was
quenched with water (20 mL), and the mixture was extracted with DCM
(2.times.50 mL). The combined organic layers were washed with brine
(100 mL), dried over Na.sub.2SO.sub.4 and filtered. The solvent was
removed under reduced pressure to afford the desired compound
Tert-butyl
2-(2-(4-(2-(benzyloxy)ethyl)piperazin-1-yl)ethoxy)acetate (1.0 g,
2.65 mmol, crude) as colorless oil. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 7.35-7.28 (m, 5H), 4.55 (s, 2H), 4.00 (s, 2H),
3.68 (t, J=5.6 Hz, 2H), 3.61 (t, J=5.6 Hz, 2H), 2.65-2.58 (m, 12H),
1.50 (s, 9H). Chemical Formula: C.sub.21H.sub.34N.sub.2O.sub.4;
Molecular Weight: 378.51
5. Step--Synthesis of
2-(2-(4-(2-(Benzyloxy)ethyl)piperazin-1-yl)ethoxy)acetic acid
##STR00627##
[1557] A solution of tert-butyl
2-(2-(4-(2-(benzyloxy)ethyl)piperazin-1-yl)ethoxy)acetate (1 g,
2.65 mmol) in HCl/dioxane (4.0 M, 5 mL) was stirred at r.t. for 3
hours. The solvent was removed under reduced pressure to afford the
desired compound
2-(2-(4-(2-(Benzyloxy)ethyl)piperazin-1-yl)ethoxy)acetic acid (0.97
g, 3.01 mmol, crude) as colorless oil. Chemical Formula:
C.sub.17H.sub.26N.sub.2O.sub.4; Molecular Weight: 322.40
6. Step--Synthesis of
2-(2-(4-(2-(Benzyloxy)ethyl)piperazin-1-yl)ethoxy)-N-(4-(isopropylamino)--
6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)pyridin-3-yl)acetamide
##STR00628##
[1559] To a solution of
2-(2-(4-(2-(Benzyloxy)ethyl)piperazin-1-yl)ethoxy)acetic acid (0.97
g, 3.01 mmol),
N.sup.4-isopropyl-N.sup.2-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)pyri-
dine-2,4,5-triamine [J. Med. Chem. 2015, 58, 8877-8895] (0.9 g,
2.50 mmol), and DIPEA (0.65 g, 5.01 mmol) in DCM (60 mL) was added
HATU (1.9 g, 5.01 mmol). The resulting solution was stirred at r.t.
for 2 h. The reaction mixture was quenched with water (20 mL), and
the mixture was extracted with DCM (2.times.50 mL). The combined
organic layers were washed with brine (100 mL), dried over
Na.sub.2SO.sub.4 and filtered. The solvent was evaporated under
reduced pressure. The residue was purified by column chromatography
(DCM:MeOH=20:1) to afford the desired compound
2-(2-(4-(2-(Benzyloxy)ethyl)piperazin-1-yl)ethoxy)-N-(4-(isopropylamino)--
6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)pyridin-3-yl)acetamide
(0.63 g, 0.95 mmol, crude) as red solid.
7. Step--Synthesis of
2-((2-(4-(2-(Benzyloxy)ethyl)piperazin-1-yl)ethoxy)methyl)-1-isopropyl-N--
(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-1H-imidazo[4,5-c]pyridin-6-ami-
ne
##STR00629##
[1561] A solution of
2-(2-(4-(2-(Benzyloxy)ethyl)piperazin-1-yl)ethoxy)-N-(4-(isopropylamino)--
6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)pyridin-3-yl)acetamide
(0.63 g, 0.95 mmol) in AcOH (5 mL) was irradiated to 150.degree. C.
with microwave for 6 h. After cooling to rt, the solvent was
removed in vacuo to afford the desired product
2-((2-(4-(2-(Benzyloxy)ethyl)piperazin-1-yl)ethoxy)methyl)-1-isopropyl-N--
(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-1H-imidazo[4,5-c]pyridin-6-ami-
ne (1.0 g, 1.56 mmol, crude) as red oil. LC-MS: (ES.sup.+):
m/z644.4 [M+H]. t.sub.R=2.749 min. Chemical Formula:
C.sub.35H.sub.49N.sub.9O.sub.3; Molecular Weight: 643.82
8. Step--Synthesis of
2-(4-(2-((1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)piperazin-1-yl)ethanol
##STR00630##
[1563] To a solution of
2-((2-(4-(2-(Benzyloxy)ethyl)piperazin-1-yl)ethoxy)methyl)-1-isopropyl-N--
(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-1H-imidazo[4,5-c]pyridin-6-ami-
ne (1 g, 1.56 mmol) in methanol (20 mL) was added Pd(OH).sub.2/C
(20%, 0.50 g) and conc. HCl (0.1 ml) subsequently at rt. The
mixture was stirred for 2 h under H.sub.2 at 1 atm. The reaction
mixture was filtered and the filtrate was concentrated under vacuum
to afford the desired product
2-(4-(2-((1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4--
yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)piperazin-1-yl)ethan-
ol (0.31 g, 0.56 mmol, crude) as red oil. LC-MS: (ES.sup.+): m/z
554.3 [M+H]. t.sub.R=2.342 min. Chemical Formula:
C.sub.28H.sub.43N.sub.9O.sub.3; Molecular Weight: 553.70
9. Step--Synthesis of Tert-butyl
5-amino-4-(5-(2-(4-(2-((1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimi-
din-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)pierazin-1-yl)-
ethoxy)-1,3-dioxoisoindolin-2-yl)-5-oxopentanoate
##STR00631##
[1565] To a solution of
2-(4-(2-((1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)piperazin-1-yl)ethanol
(55 mg, 0.10 mmol), PPh.sub.3 (131 mg, 0.51 mmol) and tert-butyl
5-amino-4-(5-hydroxy-1,3-dioxoisoindolin-2-yl)-5-oxopentanoate (70
mg, 0.21 mmol) in dry THF (5 mL) was added DIAD (101 mg, 0.51 mmol)
drop-wise at 0.degree. C. under N.sub.2. The mixture was stirred at
room temperature for 1 h. The resulting solution was quenched with
water (50 mL), and the mixture was extracted with DCM (20
mL.times.2). The combined organic layers were dried over
MgSO.sub.4, and concentrated under vacuum. The residue was purified
by chromatography (silica gel, DCM/MeOH=1/1) to afford the desired
product Tert-butyl
5-amino-4-(5-(2-(4-(2-((1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimi-
din-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)piperazin-1-yl-
)ethoxy)-1,3-dioxoisoindolin-2-yl)-5-oxopentanoate (90 mg, 0.10
mmol, crude) as yellow solid. Chemical Formula:
C.sub.45H.sub.61N.sub.11O.sub.8; Molecular Weight: 884.03
10. Step--Synthesis of
2-(2,6-Dioxopiperidin-3-yl)-5-(2-(4-(2-((1-isopropyl-6-((2-(4-methoxypipe-
ridin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)eth-
yl)piperazin-1-yl)ethoxy)isoindoline-1,3-dione
##STR00632##
[1567] To a solution of Tert-butyl
5-amino-4-(5-(2-(4-(2-((1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimi-
din-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)ethyl)piperazin-1-yl-
)ethoxy)-1,3-dioxoisoindolin-2-yl)-5-oxopentanoate (90 mg, 0.10
mmol) in CH.sub.3CN (10 mL) and was added TsOH (172 mg, 1.01 mmol)
at rt. The resulting solution was stirred at 80.degree. C. for 3 h.
After cooling to 0.degree. C., the reaction was quenched with aq
NaHCO.sub.3, and the mixture was extracted with DCM (10
mL.times.3). The combined organic layers were dried over anhydrous
sodium sulfate, filtered and concentrated under vacuum. The residue
was purified by silica gel column chromatography (DCM/MeOH=20/1) to
afford
2-(2,6-Dioxopiperidin-3-yl)-5-(2-(4-(2-((1-isopropyl-6-((2-(4-methoxypipe-
ridin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)eth-
yl)piperazin-1-yl)ethoxy)isoindoline-1,3-dione (14 mg, 0.02 mmol,
17%) as white solid. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
8.73 (br, 1H), 8.71 (s, 1H), 8.47 (s, 1H), 8.04 (d, J=5.6 Hz, 1H),
7.78 (d, J=8.4 Hz, 1H), 7.66 (s, 1H), 7.35 (s, 1H), 7.20-7.18 (m,
1H), 6.06 (d, J=5.6 Hz, 1H), 4.97-4.93 (m, 2H), 4.79 (s, 2H),
4.39-4.36 (m, 2H), 4.21-4.19 (m, 2H), 3.64-3.61 (m, 2H), 3.50-3.48
(m, 3H), 3.41 (s, 3H), 2.88-2.82 (m, 5H), 2.61-2.52 (m, 10H),
2.14-2.11 (m, 1H), 2.09-1.96 (m, 2H), 1.67-1.65 (m, 8H). LC-MS:
(ES.sup.+): m/z 811.3 [M+H]. t.sub.R=2.639 min. Chemical Formula:
C.sub.41H.sub.51N.sub.11O.sub.7; Molecular Weight: 809.91
Synthesis of Example 78
2-(2,6-dioxopiperidin-3-yl)-5-(4-(2-(1-(4-((1-isopropyl-2-methyl-1H-imidaz-
o[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)-4-methoxypiperidin-4-yl)ethyl)p-
iperazin-1-yl)isoindoline-1,3-dione
##STR00633##
##STR00634##
[1569] The synthesis of 2-(4-methoxypiperidin-4-yl)ethan-1-ol
followed the route described in Monatshefte fuer Chemie, 2004, 135
(7), 899-909
Experimental Section
1. Step--Synthesis of 6-Bromo-N4-isopropylpyridine-3,4-diamine
##STR00635##
[1571] To a solution of 2-bromo-N-isopropyl-5-nitropyridin-4-amine
(1 g, 3.85 mmol) in ethanol (30 mL) were added NH.sub.4Cl solution
(0.57 g in 10 mL water, 105.6 mmol) and Fe powder (645 mg, 11.55
mmol) subsequently. The resulting reaction was stirred at
70.degree. C. for 3 h. After cooling to rt, the mixture was
filtered through Celite, and the filtrate was concentrated under
vacuum. The residue was dissolved in sat. sodium bicarbonate
solution, and the aqueous was extracted with ethyl acetate (20
mL.times.3). The combined organic layers were dried over sodium
sulfate and concentrated to afford the desired compound
6-Bromo-N4-isopropylpyridine-3,4-diamine (920 mg, 92%), which was
used in the next reaction without further purification. .sup.1H NMR
(400 MHz, CDCl3): .delta. 7.62 (s, 1H), 6.57 (s, 1H), 4.19 (br,
1H), 3.59-3.64 (m, 1H), 3.00 (br, 2H), 1.26 (d, J=6.4 Hz, 6H).
Chemical Formula: C.sub.8H.sub.12BrN.sub.3; Molecular Weight:
230.11
2. Step--Synthesis of
6-Bromo-1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridine
##STR00636##
[1573] A solution of 6-Bromo-N4-isopropylpyridine-3,4-diamine (900
mg, 3.913 mmol), CH.sub.3COOH (235 mg, 3.913 mmol), EDCI (897 mg,
4.696 mmol), HOBt (634 mg, 4.696 mmol), and Et.sub.3N (593 mg,
5.870 mmol) in DCM (15 mL) was stirred at rt overnight. After the
reaction was quenched with water (10 mL), the mixture was extracted
with DCM (20 mL.times.2). The combined organic layers were dried
(Na.sub.2SO.sub.4), filtered and concentrated under reduced
pressure. The residue was purified by silica gel column to afford
the desired acet-amide intermediate (1.1 g, crude 100%,).
[1574] A solution of the acetamide intermediate (1.0 g, 3.67 mmol)
in CH.sub.3COOH (10 mL) was irradiated to 150.degree. C. with
microwave for 6 h. After cooling to rt, the solvent was removed
under vacuum to afford crude desired product
6-Bromo-1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridine (475 mg,
48%). .sup.1H NMR (400 MHz, CDCl3): .delta. 8.70 (s, 1H), 7.57 (s,
1H), 4.62-4.65 (m, 1H), 2.64 (s, 3H), 1.63 (d, J=6.8 Hz, 6H).
Chemical Formula: C.sub.10H.sub.12BrN.sub.3; Molecular Weight:
254.13
3. Step--Synthesis of
N-(2-Chloropyrimidin-4-yl)-1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin--
6-amine
##STR00637##
[1576] A mixture of
6-Bromo-1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridine (467 mg,
1.85 mmol), 2-chloropyrimidin-4-amine (266 mg, 2.04 mmol),
Pd.sub.2(dba).sub.3 (213 mg, 0.37 mmol), xantphos (429 mg, 0.74
mmol), and Cs.sub.2CO.sub.3 (1.8 g, 5.55 mmol) in dioxane (10 mL)
was heated to 100.degree. C. overnight in sealed tube with
sand-bath. After cooling to rt, the mixture was filtered through
Celite, and the filtered cake was washed with EtOAc. The filtrate
was concentrated under vacuum. The residue was purified by
prep-HPLC to afford the desired product
N-(2-Chloropyrimidin-4-yl)-1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin--
6-amine (150 mg, 27%).
[1577] .sup.1H NMR (400 MHz, DMSO): .delta. 8.57 (s, 1H), 8.25 (d,
J=5.6 Hz, 1H), 8.12 (br s, 1H), 7.48 (br s, 1H), 4.71-4.75 (m, 1H),
2.58 (s, 3H), 1.58 (d, J=7.2 Hz, 6H).
[1578] Chemical Formula: C.sub.14H.sub.15ClN.sub.6; Molecular
Weight: 302.77
4. Step--Synthesis of
2-(1-(4-((1-Isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)pyrimi-
din-2-yl)-4-methoxypiperidin-4-yl)ethan-1-ol
##STR00638##
[1580] A solution of
N-(2-Chloropyrimidin-4-yl)-1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin--
6-amine (103 mg, 0.341 mmol), 2-(4-methoxypiperidin-4-yl)ethan-1-ol
(49 mg, 0.31 mmol), Et.sub.3N (157 mg, 1.55 mmol) in isopropanol
was irradiated to 150.degree. C. with microwave for 20 min under
N.sub.2. The mixture was filtered through Celite and the filtrate
was concentrated under vacuum. The residue was purified by
chromatography column (PE/EA=10/1 to EA) to afford the desired
product
2-(1-(4-((1-Isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)pyrimi-
din-2-yl)-4-methoxypiperidin-4-yl)ethan-1-ol (66 mg, 50%) as a
white solid.
[1581] .sup.1H NMR (400 MHz, MeOD): .delta. 8.52 (s, 1H), 8.37 (s,
1H), 7.91 (d, J=6.0 Hz, 1H), 6.36 (d, J=6.0 Hz, 1H), 4.33 (d,
J=13.2 Hz, 2H), 3.67 (t, J=7.2 Hz, 2H), 3.30-3.35 (m, 3H), 3.26 (s,
3H), 2.64 (s, 3H), 1.89-1.92 (d, J=13.2 Hz, 2H), 1.82 (t, J=7.2 Hz,
2H), 1.67 (d, J=6.8 Hz, 6H), 1.61-1.64 (m, 2H).
[1582] Chemical Formula: C.sub.22H.sub.31N.sub.7O.sub.2; Molecular
Weight: 425.54
[1583] LC-MS (ES.sup.+): m/z 426.3 [MH.sup.+]; t.sub.R=2.51 min
##STR00639##
5. Step--Synthesis of
2-(1-(4-((1-Isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)pyrimi-
din-2-yl)-4-methoxypiperidin-4-yl)acetaldehyde
##STR00640##
[1585] To a solution of
2-(1-(4-((1-Isopropyl-2-methyl-H-imidazo[4,5-c]pyridin-6-yl)amino)pyrimid-
in-2-yl)-4-methoxypiperidin-4-yl)ethan-1-ol (180 mg, 0.391 mmol) in
DCM (10 mL) was added Dess-Martin Periodinane (332 mg, 0.782 mmol).
The resulting mixture was stirred at rt for 2 h. The resulting
solution was filtered and evaporated to afford the desired product
2-(1-(4-((1-Isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)pyrimi-
din-2-yl)-4-methoxypiperidin-4-yl)acetaldehyde (75 mg, crude),
which was used in the next reaction without further
purification.
[1586] 6. Step--Synthesis of tert-butyl
4-(2-(1-(4-((1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)pyr-
imidin-2-yl)-4-methoxypiperidin-4-yl)ethyl)piperazine-1-carboxylate
followed the reductive amination procedure as described in example
307.
##STR00641##
[1587] 7. Step--Synthesis of
1-isopropyl-N-(2-(4-methoxy-4-(2-(piperazin-1-yl)ethyl)piperidin-1-yl)pyr-
imidin-4-yl)-2-methyl-1H-imidazo[4,5-c]pyridin-6-amine using HCl in
Methanol to cleave BOC protecting group. The crude product was used
without further purification.
##STR00642##
[1588] 8. Step--Synthesis of
2-(2,6-dioxopiperidin-3-yl)-5-(4-(2-(1-(4-((1-isopropyl-2-methyl-1H-imida-
zo[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)-4-methoxypiperidin-4-yl)ethyl)-
piperazin-1-yl)isoindoline-1,3-dione followed the S.sub.NAr
reaction conditions described in example 71.
##STR00643##
[1589] .sup.1HNMR (400 MHz, DMSO-d.sub.6): .delta.: 11.08 (s, 1H),
9.69 (s, 1H), 8.50 (s, 1H), 8.40 (s, 1H), 7.95 (d, J=5.6 Hz, 1H),
7.67 (d, J=7.2 Hz, 1H), 7.33 (s, 1H), 7.25 (d, J=7.2 Hz, 1H), 6.39
(d, J=4.0 Hz, 1H), 5.04-5.09 (m, 1H), 4.71 (t, J=6.4 Hz, 1H), 4.34
(d, J=12.4 Hz, 2H), 3.42 (s, 4H), 3.21-3.24 (m, 3H), 3.15 (s, 3H),
3.31 (s, 3H), 2.55 (s, 3H), 2.37 (s, 3H), 1.98-2.02 (m, 2H),
1.71-1.78 (m, 4H), 1.51 (s, 6H).
[1590] LCMS: m/e=375.8=[M+2H].sup.2+, t.sub.R=2.60 min.
[1591] Chemical Formula: C.sub.39H.sub.47N.sub.11O.sub.5; Molecular
Weight: 749.9
Synthesis of Example 80
##STR00644##
[1592]
(2S,4R)-1-((2S)-2-(2-(4-(5-(2-(1-(5-fluoro-2-hydroxyphenyl)-2-oxo-2-
-(thiazol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)pent-4-yn-1-yl)piperazin-1-
-yl)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N--((S)-1-(4-(4-methylthiaz-
ol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00645##
[1593] Experiments
1. Step--Synthesis of
2-amino-2-(2-(benzyloxy)-5-fluorophenyl)acetonitrile
##STR00646##
[1595] A mixture of 2-(benzyloxy)-5-fluorobenzaldehyde (80 g, 347
mmol) in ammonia solution (7.0 M in methanol, 500 ml) was stirred
at 0.degree. C. for 0.5 hour, then trimethylsilyl cyanide (37.2 g,
375 mmol) was added dropwise at the same temperature. The resulting
reaction mixture was stirred at 45.degree. C. for 3 hour. TLC
showed the reaction was complete. The volatiles were evaporated
under reduced pressure to give
2-amino-2-(2-(benzyloxy)-5-fluorophenyl)acetonitrile (crude) as
yellow oil which was used in next step without further
purification.
2. Step--Synthesis of methyl
2-amino-2-(2-(benzyloxy)-5-fluorophenyl)acetate hydrochloride
##STR00647##
[1597] Thionyl chloride (60 ml) was added to methanol (400 ml)
dropwise at 0.degree. C. and the resulting reaction mixture was
stirred at the same temperature for 10 min. to afford crude
solution of hydrogen chloride in methanol. The mixture of hydrogen
chloride in methanol (400 ml) and
2-amino-2-(2-(benzyloxy)-5-fluorophenyl)acetonitrile (crude, 347
mmol) was stirred at 65.degree. C. overnight. TLC showed the
reaction was complete. The volatiles were removed under reduced
pressure. The residue was dissolved in methanol (50 ml), and ethyl
acetate (500 ml) was added. The resulting mixture was stirred at
room temperature for 0.5 hour, and white solid precipitated. The
solid was collected by filtration and dried under vacuum to afford
methyl 2-amino-2-(2-(benzyloxy)-5-fluorophenyl)acetate
hydrochloride (82 g, yield: 72% over 2 steps) as white solid.
LC_MS: (ES.sup.+): m/z 290.2 [M+H].sup.+. t.sub.R=1.852 min. 1H NMR
(400 Hz, CD.sub.3OD): .delta. 3.73 (s, 3H), 5.22-5.24 (m, 2H), 5.35
(s, 1H), 7.19-7.26 (m, 3H), 7.36-7.48 (m, 5H). Chemical Formula:
C.sub.16H.sub.17ClFNO.sub.3; Molecular Weight: 289.30;
3. Step--Synthesis of 2-iodo-6-methylbenzoic acid
##STR00648##
[1599] A mixture of 2-methylbenzoic acid (10.0 g, 73.5 mmol),
N-Iodosuccinimide (16.0 g, 73.5 mmol) and palladium diacetate (1.6
g, 7.35 mmol) in N,N-dimethylformamide (160 ml) was stirred at
100.degree. C. for 2 hours under nitrogen atmosphere. TLC showed
the reaction was complete. The cooled reaction mixture was allowed
to cool to room temperature and partitioned between ethyl acetate
(300 ml) and water (500 ml). The organic layer was collected, and
the aqueous layer was extracted with ethyl acetate (100
ml.times.2). The combined organic layers were washed with brine
(200 ml), dried over anhydrous sodium sulfate, and concentrated
under reduced pressure to give 2-iodo-6-methylbenzoic acid (16 g,
yield 84%) as yellow solid which was used in next step directly
without further purification. 1H NMR (400 Hz, CDCl.sub.3): .delta.
2.29 (s, 3H), 7.06 (t, J=8.0 Hz, 1H), 7.27 (d, J=7.6 Hz, 1H), 7.67
(d, J=8.0 Hz, 1H). Chemical Formula: C.sub.8H.sub.7IO.sub.2;
Molecular Weight: 262.04;
4. Step--Synthesis of methyl 2-iodo-6-methylbenzoate
##STR00649##
[1601] A mixture of 2-iodo-6-methylbenzoic acid (5.6 g, 21.4 mmol),
iodomethane (2.5 ml, 40.1 mmol) and potassium carbonate (5.0 g,
35.9 mmol) in N,N-dimethylformamide (40 ml) was a stirred at room
temperature overnight. TLC showed the reaction was complete. The
reaction mixture was partitioned between ethyl acetate (50 ml) and
water (50 ml). The organic layer was collected, and the aqueous
layer was extracted with ethyl acetate (100 ml.times.2). The
combined organic layers were washed with brine (200 ml), dried over
anhydrous sodium sulfate, and concentrated under reduced pressure
to afford a crude residue which was purified by silica gel flash
chromatography (eluted with 5% ethyl acetate in hexane) to afford
methyl 2-iodo-6-methylbenzoate (3.8 g, yield 63%) as white solid.
LC_MS: (ES.sup.+): m/z 277.0 [M+H].sup.+. t.sub.R=2.863 min. 1H NMR
(400 Hz, CDCl.sub.3): .delta. 2.35 (s, 3H), 3.97 (s, 3H), 7.01 (t,
J=8.0 Hz, 1H), 7.20 (d, J=7.6 Hz, 1H), 7.67 (d, J=8.0 Hz, 1H).
Chemical Formula: C.sub.9H.sub.9IO.sub.2; Molecular Weight:
276.07;
5. Step--Synthesis of methyl 2-(bromomethyl)-6-iodobenzoate
##STR00650##
[1603] A mixture of methyl 5-iodo-2-methylbenzoate (3.8 g, 13.7
mmol), AIBN (2,2'-Dimethyl-2,2'-azodipropionitrile) (1 g, 6.16
mmol) and N-Bromosuccinimide (2.9 g, 16.5 mmol) in carbon
tetrachloride (40 ml) was refluxed overnight. TLC showed the
reaction was complete. The cooled reaction mixture was partitioned
between dichloromethane (30 ml) and water (20 ml). The organic
layer was collected, and the aqueous layer was extracted with
dichloromethane (30 ml.times.2). The combined organic layers were
washed with brine (20 ml), dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to give a crude residue which
was purified by silica gel flash chromatography (eluted with 5%
ethyl acetate in hexane) to afford methyl
2-(bromomethyl)-6-iodobenzoate (2.4 g, yield 50%) as white solid.
1H NMR (400 Hz, CDCl.sub.3): .delta. 3.99 (s, 3H), 4.78 (s, 2H),
7.09 (t, J=8.0 Hz, 1H), 7.40 (d, J=7.6 Hz, 1H), 7.80 (d, J=8.0 Hz,
1H). Chemical Formula: C.sub.9H.sub.8BrIO.sub.2; Molecular Weight:
354.97;
6. Step--Synthesis of methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-iodo-1-oxoisoindolin-2-yl)acetate
##STR00651##
[1605] A mixture of methyl methyl
2-amino-2-(2-(benzyloxy)-5-fluorophenyl)acetate (1.2 g, 4.1 mmol),
methyl 2-(bromomethyl)-6-iodobenzoate (1.5 g, 4.1 mmol) and
triethylamine (0.9 ml, 6.2 mmol) in toluene (10 ml) was stirred at
110.degree. C. overnight under nitrogen atmosphere. TLC showed the
reaction was complete. The cooled reaction mixture was partitioned
between ethyl acetate (50 ml) and water (30 ml). The organic layer
was collected, and the aqueous layer was extracted with ethyl
acetate (30 ml.times.2). The combined organic layers were washed
with brine (30 ml), dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to give a crude residue which
was purified by silica gel flash chromatography (eluted with 10%
ethyl acetate in hexane) to afford methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-iodo-1-oxoisoindolin-2-yl)acetate
(1.3 g, yield: 59%) as white solid. LC_MS: (ES.sup.+): m/z 532.1
[M+H].sup.+. t.sub.R=3.137 min. 1H NMR (400 Hz, CD.sub.3OD):
.delta. 3.62 (s, 3H), 3.86 (d, J=17.2 Hz, 1H), 4.58 (d, J=16.2 Hz,
1H), 5.04-5.15 (m, 2H), 6.43 (s, 1H), 6.93-6.97 (m, 1H), 7.00-7.05
(m, 1H), 7.06-7.09 (m, 1H), 7.19 (t, J=7.6 Hz, 1H), 7.27-7.39 (m,
6H), 7.89 (d, J=7.6 Hz, 1H). Chemical Formula: C24H19FINO4;
Molecular Weight: 531.31; See also the closely related synthesis of
example 307.
##STR00652##
7. Step--Synthesis of tert-butyl
4-(pent-4-yn-1-yl)piperazine-1-carboxylate
##STR00653##
[1607] To a stirred solution of pent-4-yn-1-yl
4-methylbenzenesulfonate (2.88 mg, 12.09 mmol),
N-ethyl-N-isopropylpropan-2-amine (3.12 g, 24.17 mmol) and
potassium iodide (201 mg, 1.21 mmol) in N,N-dimethylformamide (30
ml) was added tert-butyl piperazine-1-carboxylate (2.25 mg, 12.09
mmol), and the mixture was stirred at 50.degree. C. overnight under
nitrogen. TLC showed the reaction was complete. The reaction
mixture was partitioned between ethyl acetate (50 ml) and water (50
ml), the organic layer was washed with brine (50 ml.times.2), dried
over anhydrous sodium sulfate and concentrated under reduced
pressure to give a crude residue which was purified by silica gel
flash column chromatography (eluted with 20-40% ethyl acetate in
hexane) to afford tert-butyl
4-(pent-4-yn-1-yl)piperazine-1-carboxylate (1.7 g, yield 56%) as
yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 1.46 (s,
9H), 1.68-1.75 (m, 2H), 1.95 (t, J=2.6 Hz, 1H), 2.22-2.26 (m, 2H),
2.38 (t, J=4.8 Hz, 4H), 2.42-2.46 (m, 2H), 3.42 (t, J=5.0 Hz, 4H).
Chemical Formula: C.sub.14H.sub.24N.sub.2O.sub.2; Molecular Weight:
252.35;
8. Step--Synthesis of tert-butyl
4-(5-(2-(1-(2-(benzyloxy)-5-fluorophenyl)-2-oxo-2-(thiazol-2-ylamino)ethy-
l)-3-oxoisoindolin-4-yl)pent-4-yn-1-yl)piperazine-1-carboxylate
##STR00654##
[1609] To a solution of
2-(5-(benzyloxy)-2-fluorophenyl)-2-(7-iodo-1-oxoisoindolin-2-yl)-N-(thiaz-
ol-2-yl)acetamide (468 mg, 0.78 mmol),
1-methyl-4-(pent-4-yn-1-yl)piperazine (296 mg, 1.17 mmol)and
triethylamine (474 mg, 4.68 mmol) in N,N-dimethylformamide (8 ml)
was added cuprous iodide (30 mg, 0.16 mmol) and
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (109
mg, 0.16 mmol) at room temperature under nitrogen atmosphere; the
mixture was degassed with nitrogen three times. The resulting
mixture was refluxed for 2 hours. TLC showed the reaction was
complete. The reaction mixture was partitioned between ethyl
acetate (30 ml) and water (20 ml); the organic layer was collected
and the aqueous layer was extracted with ethyl acetate (20
ml.times.2). The combined organic layers were washed with brine (40
ml), dried over anhydrous sodium sulfate, and concentrated under
reduced pressure to give a crude residue which was purified by
silica gel flash column chromatography (eluted with 5-10% methanol
in dichloromethane) to afford tert-butyl
4-(5-(2-(1-(2-(benzyloxy)-5-fluorophenyl)-2-oxo-2-(thiazol-2-ylamino)ethy-
l)-3-oxoisoindolin-4-yl)pent-4-yn-1-yl)piperazine-1-carboxylate
(348 mg, yield 62%) as black solid. LC_MS: (ES.sup.+): m/z 724.3
[M+H].sup.+. t.sub.R=2.483 min. Chemical Formula:
C.sub.40H.sub.42FN.sub.5O.sub.5S; Molecular Weight: 723.86;
9. Step--Synthesis of
2-(2-(benzyloxy)-5-fluorophenyl)-2-(1-oxo-7-(5-(piperazin-1-yl)pent-1-yn--
1-yl)isoindolin-2-yl)-N-(thiazol-2-yl)acetamide TFA salt
##STR00655##
[1611] To a stirred solution of tert-butyl
4-(5-(2-(1-(2-(benzyloxy)-5-fluorophenyl)-2-oxo-2-(thiazol-2-ylamino)ethy-
l)-3-oxoisoindolin-4-yl)pent-4-yn-1-yl)piperazine-1-carboxylate
(348 mg, 0.48 mmol) in dichloromethane (3 ml) was added
2,2,2-trifluoroacetic acid (2 ml) at 0.degree. C., the mixture
solution was stirred at this temperature for 2 hours. TLC showed
the reaction was complete. The volatiles were evaporated under
reduced pressure to afford
2-(2-(benzyloxy)-5-fluorophenyl)-2-(1-oxo-7-(5-(piperazin-1-yl)pent-1-yn--
1-yl)isoindolin-2-yl)-N-(thiazol-2-yl)acetamide TFA salt (crude) as
yellow oil. LC_MS: (ES.sup.+): m/z 624.3 [M+H].sup.+. t.sub.R=1.958
min. Chemical Formula: C.sub.35H.sub.34FN.sub.5O.sub.3S; Molecular
Weight: 623.74;
10. Step--Synthesis of
(2S,4R)-1-((2S)-2-(2-(4-(5-(2-(1-(2-(benzyloxy)-5-fluorophenyl)-2-oxo-2-(-
thiazol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)pent-4-yn-1-yl)piperazin-1-y-
l)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N--((S)-1-(4-(4-methylthiazol-
-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00656##
[1613] To a stirred solution of
2-(2-(benzyloxy)-5-fluorophenyl)-2-(1-oxo-7-(5-(piperazin-1-yl)pent-1-yn--
1-yl)isoindolin-2-yl)-N-(thiazol-2-yl)acetamide (crude, <=0.48
mmol), N-ethyl-N-isopropylpropan-2-amine (124 mg, 0.96 mmol) and
potassium iodide (8 mg, 0.05 mmol) in N,N-dimethylformamide (3 ml)
was added
(2S,4R)-1-((S)-2-(2-chloroacetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N--(-
(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
(250 mg, 0.48 mmol), and the mixture was stirred at 50.degree. C.
overnight under nitrogen. TLC showed the reaction was complete. The
reaction mixture was partitioned between ethyl acetate (20 ml) and
water (20 ml), the organic layer was washed with brine (20
ml.times.2), dried over anhydrous sodium sulfate and concentrated
under reduced pressure to give a crude residue which was purified
by silica gel flash column chromatography (eluted with 2-10%
methanol in dichloromethane) to afford
(2S,4R)-1-((2S)-2-(2-(4-(5-(2-(1-(2-(benzyloxy)-5-fluorophenyl)-2-oxo-2-(-
thiazol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)pent-4-yn-1-yl)piperazin-1-y-
l)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N--((S)-1-(4-(4-methylthiazol-
-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (90 mg, yield 17%) as
yellow oil. LC_MS: (ES+): m/z 1108.4 [M+H]+. t.sub.R=2.458 min.
Chemical Formula: C.sub.60H.sub.66FN.sub.9O.sub.7S.sub.2; Molecular
Weight: 1108.35;
11. Step--Synthesis of
(2S,4R)-1-((2S)-2-(2-(4-(5-(2-(1-(2-fluoro-5-hydroxyphenyl)-2-oxo-2-(thia-
zol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)pent-4-yn-1-yl)piperazin-1-yl)ac-
etamido)-3,3-dimethylbutanoyl)-4-hydroxy-N--((S)-1-(4-(4-methylthiazol-5-y-
l)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00657##
[1615] To a stirred solution of
(2S,4R)-1-((2S)-2-(2-(4-(5-(2-(1-(2-(benzyloxy)-5-fluorophenyl)-2-oxo-2-(-
thiazol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)pent-4-yn-1-yl)piperazin-1-y-
l)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N--((S)-1-(4-(4-methylthiazol-
-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (90 mg, 0.08 mmol) in
anhydrous dichloromethane (5 ml) was added boron tribromide (102
mg, 0.41 mmol) in dichloromethane (1 ml) dropwise at -40.degree. C.
under nitrogen. The mixture solution was stirred at this
temperature for 2 hours. TLC showed the reaction was complete. The
reaction mixture was quenched with water (3 ml) at -60.degree. C.
and aqueous solution of sodium bicarbonate was added till pH 7-8.
The mixture was diluted with dichloromethane (20 ml), the organic
layer was washed with brine (20 ml.times.2), dried over anhydrous
sodium sulfate and concentrated under reduced pressure to give a
crude residue which was purified by preparative TLC (eluted with 5%
methanol in dichloromethane) to afford
(2S,4R)-1-((2S)-2-(2-(4-(5-(2-(1-(5-fluoro-2-hydroxyphenyl)-2-oxo-2-(thia-
zol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)pent-4-yn-1-yl)piperazin-1-yl)ac-
etamido)-3,3-dimethylbutanoyl)-4-hydroxy-N--((S)-1-(4-(4-methylthiazol-5-y-
l)phenyl)ethyl)pyrrolidine-2-carboxamide (5.6 mg, yield 13% two
steps) as white solid. LC_MS: (ES.sup.+): m/z 1018.4 [M+H].sup.+.
t.sub.R=2.402 min. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
0.96 (s, 9H), 1.34-1.36 (m, 3H), 1.45-1.51 (m, 1H), 1.74-1.83 (m,
1H), 1.96-2.06 (m, 2H), 2.11-2.21 (m, 2H), 2.44-2.45 (m, 3H),
3.00-3.16 (m, 6H), 3.61-3.62 (m, 1H), 3.67-3.73 (m, 2H), 3.81-3.98
(m, 4H), 4.23-4.29 (m, 1H), 4.42-4.46 (m, 1H), 4.52-4.65 (m, 2H),
4.88-4.93 (m, 1H), 5.14 (s, 1H), 6.32 (s, 1H), 6.84-6.87 (m, 1H),
6.99-7.02 (m, 1H), 7.13 (s, 2H), 7.26 (s, 2H), 7.36-7.42 (m, 4H),
7.47-7.49 (m, 1H), 7.52-7.54 (m, 1H), 7.59-7.61 (m, 1H), 7.66-7.70
(m, 1H), 7.81-7.83 (m, 1H), 8.18 (s, 1H), 8.39-8.40 (m, 1H), 8.98
(s, 1H), 10.13 (s, 1H), 12.64 (s, 1H). Chemical Formula:
C.sub.53H.sub.60FN.sub.9O.sub.7S.sub.2; Molecular Weight:
1018.23;
Synthesis of Example 81
(2S,4R)-1-((S)-18-(tert-butyl)-1-(1-isopropyl-6-((2-(4-methoxypiperidin-1--
yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)-16-oxo-2,5,8,11,14-
-pentaoxa-17-azanonadecan-19-oyl)-4-hydroxy-N--((S)-1-(4-(4-methylthiazol--
5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00658##
[1617] Synthesis Scheme:
##STR00659##
Experimental Section
1. Step--Synthesis of
N-(4-(Isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)pyridin-3-yl)-1-phenyl-2,5,8,11,14,17-hexaoxanonadecan-19-amide
##STR00660##
[1619] To a solution of
N.sup.4-isopropyl-N.sup.2-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)pyri-
dine-2,4,5-triamine [J. Med. Chem. 2015, 58, 8877-8895] (580 mg,
1.62 mmol) in DCM (10 mL) were added
1-phenyl-2,5,8,11,14,17-hexaoxanonadecan-19-oic acid (689.7 mg,
1.78 mmol, see example 70), TEA (328.4 mg, 3.25 mmol), EDCI (465.8
mg, 2.43 mmol) and HOBt (328.4 mg, 2.43 mmol) at 10.degree. C. The
resulting solution was stirred at 20.degree. C. for 16 h. The
reaction was diluted with a solution of H.sub.2O (40 mL), the
mixture was extracted with DCM (20 mL.times.2). The combined
organic layers were dried over anhydrous sodium sulfate and
concentration. The residue was purified with silica gel column to
afford the desired product
N-(4-(Isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)pyridin-3-yl)-1-phenyl-2,5,8,11,14,17-hexaoxanonadecan-19-amide
(480 mg, 41% yield). Chemical Formula:
C.sub.37H.sub.55N.sub.7O.sub.8; Molecular Weight: 725.87
2. Step--Synthesis of
1-Isopropyl-N-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-2-(18-phenyl-2,-
5,8,11,14,17-hexaoxaoctadecyl)-1H-imidazo[4,5-c]pyridin-6-amine
##STR00661##
[1621] A mixture of
N-(4-(Isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino-
)pyridin-3-yl)-1-phenyl-2,5,8,11,14,17-hexaoxanonadecan-19-amide
(480 mg, 0.66 mmol) in CH.sub.3COOH (5 mL) was radiated at
120.degree. C. for 10 h with microwave. After cooling to rt, the
solvent was removed in vacuo to afford crude desired product
1-Isopropyl-N-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-2-(18-phenyl-2,-
5,8,11,14,17-hexaoxaoctadecyl)-1H-imidazo[4,5-c]pyridin-6-amine
(900 mg, crude), which was used in the next step without further
purification. Chemical Formula: C.sub.37H.sub.53N.sub.7O.sub.7;
Molecular Weight: 707.86
3. Step--Synthesis of
1-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-im-
idazo[4,5-c]pyridin-2-yl)-2,5,8,11,14-pentaoxahexadecan-16-ol
##STR00662##
[1623] To a solution of
1-Isopropyl-N-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)-2-(18-phenyl-2,-
5,8,11,14,17-hexaoxaoctadecyl)-1H-imidazo[4,5-c]pyridin-6-amine
(900 mg, crude) in MeOH (50 mL) was added Pd(OH).sub.2/C (180 mg)
at 15.degree. C. The mixture was stirred at 15.degree. C. for 2 h
under H.sub.2 1 atm. Then the mixture was filtered through Celite,
and the filtrate was concentrated to afford the desired product
1-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-im-
idazo[4,5-c]pyridin-2-yl)-2,5,8,11,14-pentaoxahexadecan-16-ol (350
mg, crude), which was used into next reaction without further
purification. Chemical Formula: C.sub.3H.sub.47N.sub.7O.sub.7;
Molecular Weight: 617.74
4. Step--Synthesis of
1-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-im-
idazo[4,5-c]pyridin-2-yl)-2,5,8,11,14-pentaoxahexadecan-16-oic
acid
##STR00663##
[1625] To a solution of
1-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-im-
idazo[4,5-c]pyridin-2-yl)-2,5,8,11,14-pentaoxahexadecan-16-ol (130
mg, crude) in DMF (5 mL) was added Jones reagent (2.0 mL) at
0.degree. C. under N.sub.2 1 atm. The mixture was stirred at
15.degree. C. for 2 h. Then the mixture was filtered through
Celite, and the filtrate was concentrated to afford the desired
product
1-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-im-
idazo[4,5-c]pyridin-2-yl)-2,5,8,11,14-pentaoxahexadecan-16-oic acid
(210 mg crude), which was used in the next reaction without further
purification. Chemical Formula: C.sub.30H.sub.45N.sub.7O.sub.8;
Molecular Weight: 631.72
5. Step--Synthesis of
(2S,4R)-1-((S)-18-(Tert-butyl)-1-(1-isopropyl-6-((2-(4-methoxypiperidin-1-
-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)-16-oxo-2,5,8,11,1-
4-pentaoxa-17-azanonadecan-19-oyl)-4-hydroxy-N--((S)-1-(4-(4-methylthiazol-
-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00664##
[1627] To a solution of
1-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-im-
idazo[4,5-c]pyridin-2-yl)-2,5,8,11,14-pentaoxahexadecan-16-oic acid
(210 mg crude, 0.25 mmol) and
(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N--((S)-1-(4-(4-me-
thylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (110 mg,
0.25 mmol) in DMF (15 mL) were added DIPEA (85 mg, 0.66 mmol) and
HATU (251 mg, 0.66 mmol) at 15.degree. C. After stirring at
15.degree. C. for 16 h, the reaction was diluted with H.sub.2O (30
mL). The resulting mixture was extracted with DCM (20 mL.times.2).
The combined organic layers were dried over anhydrous sodium
sulfate and concentration. The residue was purified with silica gel
column and prep-HPLC to afford the desired product
(2S,4R)-1-((S)-18-(Tert-butyl)-1-(1-isopropyl-6-((2-(4-methoxypip-
eridin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)-16-oxo-2,-
5,8,11,14-pentaoxa-17-azanonadecan-19-oyl)-4-hydroxy-N--((S)-1-(4-(4-methy-
lthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (4.3 mg) as a
white solid. .sup.1H NMR (400 MHz, CD.sub.3OD): .delta.: 8.77 (s,
1H), 8.60 (s, 1H), 8.45 (s, 1H), 8.24 (br.s, 1H), 7.77 (d, J=4.0
Hz, 1H), 7.52 (d, J=12.0 Hz, 1H), 7.28-7.33 (m, 4H), 4.88-4.94 (m,
3H), 4.76 (d, J=12.0 Hz, 1H), 4.56-4.59 (m, 1H), 4.47 (s, 1H),
3.91-4.00 (m, 2H), 3.51-3.66 (m, 20H), 3.31 (s, 3H), 2.37 (s, 3H),
2.09-2.16 (m, 2H), 1.86-1.89 (m, 3H), 1.60 (s, 1H), 1.58 (s, 1H),
1.39 (d, J=8.0 Hz, 3H), 0.93 (s, 9H). LCMS: m/e=529.8
[M+2H].sup.2+, t.sub.R=3.35 min. Chemical Formula:
C.sub.53H.sub.75N.sub.11O.sub.10S; Molecular Weight: 1058.3
Synthesis of Example 82
2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)am-
ino)ethoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-1-oxoisoindolin-2-yl)-2-(5-fluoro-
-2-hydroxyphenyl)-N-(thiazol-2-yl)acetamide
##STR00665##
[1628] Synthetic Scheme
##STR00666##
[1629] 1. Step--Synthesis of
2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethanol
##STR00667##
[1631] To a stirred solution of potassium tert-butanolate (38 ml,
38 mmol, 1M in tetrahydrofuran) in tetrahydrofuran (100 ml) was
added 2,2'-(ethane-1,2-diylbis(oxy))diethanol (10.6 g, 100 mmol)
slowly at room temperature, and the reaction mixture was stirred at
room temperature for 30 minutes. The reaction mixture was cooled to
0.degree. C., then 3-bromoprop-1-yne (3.6 g, 30 mmol) in
tetrahydrofuran (25 ml) was added dropwise, and the resulting
reaction mixture was allowed to warm up to room temperature and
stirred overnight. TLC showed the reaction was complete. The
reaction mixture was quenched with water (50 ml) at 0.degree. C.
and extracted with ethyl acetate (100 ml.times.2). The combined
organic layers were washed with brine (50 ml.times.2), dried over
anhydrous sodium sulfate, and concentrated under reduced pressure
to give a residue which was purified by silica gel flash
chromatography (eluted with 30% ethyl acetate in hexane) to afford
2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethanol (4 g, yield 70%)
as colorless oil. .sup.1H NMR (400 Hz, CDCl.sub.3): .delta. 2.44
(d, J=2.0 Hz, 1H), 2.56-2.86 (m, 1H), 3.60-3.63 (m, 2H), 3.66-3.75
(m, 10H), 4.21 (d, J=2.4 Hz, 2H). Chemical Formula:
C.sub.9H.sub.16O.sub.4; Molecular Weight: 188.22;
2. Step--Synthesis of
2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethyl
4-methylbenzenesulfonate
##STR00668##
[1633] To a stirred solution of
2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethanol (1.0 g, 5.3 mmol),
triethylamine (1.5 ml, 10 mmol) in dichloromethane (10 ml) was
added to sylchloride (1.1 g, 5.8 mmol) and 4-dimethylaminopyridine
(64 mg, 0.53 mmol) at 0.degree. C. The resulting solution was
allowed to warm up to room temperature and stirred for 2 hours. TLC
showed the reaction was complete. The mixture was poured into water
(20 ml) and extracted with dichloromethane (20 ml.times.2). The
combined organic layers were washed with brine (20 ml), dried over
anhydrous sodium sulfate, and concentrated under reduced pressure
to give a crude residue which was purified by silica gel flash
column chromatography (eluted with 30% ethyl acetate in hexane) to
afford 2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethyl
4-methylbenzenesulfonate (1 g, yield 55%) as colorless oil.
3. Step--Synthesis of N,N-bis-Boc
2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethan-1-amine
##STR00669##
[1635] A mixture of di-tert-butyl iminodicarboxylate (0.7 g, 3.2
mmol), 2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethyl
4-methylbenzenesulfonate (1.0 g, 2.9 mmol) and cesium carbonate
(1.15 g, 3.5 mmol) in N,N-dimethylformamide (10 ml) was stirred at
90.degree. C. overnight under nitrogen atmosphere. TLC showed the
reaction was complete. The mixture was partitioned between ethyl
acetate (30 ml) and water (30 ml). The organic layer was collected,
washed with brine (20 ml), dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to give a crude residue which
was purified by silica gel flash chromatography (eluted with 25%
ethyl acetate in hexane) to afford N,N-bis-Boc
2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethan-1-amine (550 mg,
yield 48%) as yellow oil. .sup.1H NMR (400 Hz, CDCl.sub.3): .delta.
1.50 (s, 18H), 2.42 (d, J=2.0 Hz, 1H), 2.59-3.62 (m, 6H), 3.65-3.71
(m, 4H), 3.79 (t, J=6.0 Hz, 2H), 4.20 (d, J=2.4 Hz, 2H). Chemical
Formula: C.sub.19H.sub.33NO.sub.7; Molecular Weight: 387.47;
4. Step--Synthesis of
2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethanamine
##STR00670##
[1637] A mixture of N,N-bis-Boc
2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethan-1-amine (220 mg,
0.56 mmol) and 2,2,2-trifluoroacetic acid (3 ml) in dichloromethane
(5 ml) was stirred at room temperature for 1 hour. TLC showed the
reaction was complete. The volatiles were evaporated under reduced
pressure. The residue was taken up in ethyl acetate (10 ml) and
washed with aqueous sodium bicarbonate solution (sat, 10 ml). The
organic layer was collected, dried over anhydrous sodium sulfate,
and concentrated under reduced pressure to give a residue which was
purified by silica gel flash chromatography (eluted with 5%
methanol in dichloromethane) to afford
2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethanamine (120 mg, yield
85%) as yellow oil.
5. Step--Synthesis of
2-(2,6-dioxopiperidin-3-yl)-5-((2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)-
ethyl)amino)isoindoline-1,3-dione
##STR00671##
[1639] A mixture of
2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethanamine (120 mg, 0.31
mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione
(86 mg, 0.31 mmol), and N-ethyl-N-isopropylpropan-2-amine (0.09 ml,
0.5 mmol) in 1-methylpyrrolidin-2-one (2 ml) was stirred at
90.degree. C. overnight under nitrogen. TLC showed the reaction was
complete. The reaction mixture was partitioned between ethyl
acetate (50 ml) and water (30 ml). The organic layer was collected,
washed with brine (30 ml), dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to give a crude residue which
was purified by silica gel flash chromatography (eluted with 50%
ethyl acetate in dichloromethane) to afford
2-(2,6-dioxopiperidin-3-yl)-5-((2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)-
ethyl)amino)isoindoline-1,3-dione (55 mg, yield 20%) as yellow
solid. LC_MS: (ES.sup.+): m/z 444.2 [M+H].sup.+. t.sub.R=2.135
min.
6. Step--Synthesis of
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-
-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-
-1-oxoisoindolin-2-yl)-N-(thiazol-2-yl)acetamide
##STR00672##
[1641] To a solution of
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-iodo-1-oxoisoindolin-2-yl)-N-(thiaz-
ol-2-yl)acetamide (61 mg, 0.10 mmol),
2-(2,6-dioxopiperidin-3-yl)-5-((2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)-
ethyl)amino)isoindoline-1,3-dione (46 mg, 0.10 mmol) and
triethylamine (61 mg, 0.6 mmol) in N,N-dimethylformamide (2 ml)
were added cuprous iodide (4 mg, 0.02 mmol) and
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (14 mg,
0.02 mmol) at room temperature under nitrogen atmosphere; the
mixture was degassed with nitrogen three times. The resulting
mixture was refluxed for 2 hours. TLC showed the reaction was
complete. The reaction mixture was partitioned between ethyl
acetate (20 ml) and water (10 ml). The organic layer was washed
with brine (10 ml), dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to give a crude residue which
was purified by purified by preparative TLC (eluted with 5%
methanol in dichloromethane) to afford
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-
-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-
-1-oxoisoindolin-2-yl)-N-(thiazol-2-yl)acetamide (25 mg, yield:
22%) as yellow solid. LC_MS: (ES.sup.+): m/z 915.3 [M+H].sup.+.
t.sub.R=2.840 min.
7. Step--Synthesis of
2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)a-
mino)ethoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-1-oxoisoindolin-2-yl)-2-(5-fluor-
o-2-hydroxyphenyl)-N-(thiazol-2-yl)acetamide
##STR00673##
[1643] To a stirred solution of
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-
-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-
-1-oxoisoindolin-2-yl)-N-(thiazol-2-yl)acetamide (25 mg, 0.027
mmol) in anhydrous dichloromethane (5 ml) was added boron
tribromide (67 mg, 0.27 mmol in anhydrous dichloromethane (1 ml))
dropwise at -60.degree. C. under nitrogen. The mixture solution was
stirred at -60.degree. C. for 2 hours. TLC showed the reaction was
complete. The reaction mixture was quenched with water (3 ml) at
-60.degree. C. and aqueous solution of sodium bicarbonate was added
till pH 7-8. The mixture was diluted with dichloromethane (20 ml),
the organic layer was washed with brine (20 ml.times.2), dried over
anhydrous sodium sulfate and concentrated under reduced pressure to
give a crude residue which was purified by preparative TLC (eluted
with 10% methanol in dichloromethane) to afford
2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)a-
mino)ethoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-1-oxoisoindolin-2-yl)-2-(5-fluor-
o-2-hydroxyphenyl)-N-(thiazol-2-yl)acetamide (6.5 mg, yield 28%) as
yellow solid. LC_MS: (ES.sup.+): m/z 825.5 [M+H].sup.+.
t.sub.R=2.580 min. 1H NMR (400 Hz, DMSO-d6): .delta. 1.41-1.47 (m,
1H) 1.92-2.00 (m, 4H), 2.33 (s, 1H), 2.58-2.68 (m, 1H), 2.83-2.91
(m, 1H), 3.55-3.58 (m, 6H), 3.66-3.76 (m, 2H), 3.91 (d, J=17.6 Hz,
1H), 4.43 (s, 2H), 4.56 (d, J=18.0 Hz, 1H), 5.00-5.05 (m, 1H),
5.31-5.33 (m, 1H), 6.27 (s, 1H), 6.83-6.92 (m, 2H), 6.99 (s, 1H),
7.10-7.41 (m, 4H), 7.47-7.58 (m, 4H), 9.97 (s, 1H), 11.06 (s, 1H),
12.60 (s, 1H). Chemical Formula: C.sub.41H.sub.37FN.sub.6O.sub.10S;
Molecular Weight: 824.83;
Synthesis of Example 85
(2S,4R)-4-hydroxy-1-((S)-2-(2-(3-(4-(3-((1-isopropyl-6-((2-(4-methoxypiper-
idin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)prop-
yl)piperazin-1-yl)propoxy)acetamido)-3,3-dimethylbutanoyl)-N--((S)-1-(4-(4-
-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00674##
[1645] Synthetic Scheme
##STR00675##
Experimental Section
1. Step--Synthesis of Di-tert-butyl 2,
2'-((piperazine-1,4-diylbis(propane-3,1-diyl))bis(oxy))diacetate
##STR00676##
[1647] To a solution of piperazine (86 mg, 1.0 mmol) in CH.sub.3CN
(10 mL) were added tert-butyl 2-(3-chloropropoxy)acetate (628 mg,
3.0 mmol), DBU (456 mg, 3.0 mmol) and KI (33 mg, 0.2 mmol). The
mixture was stirred at 100.degree. C. overnight. After the mixture
was cooled to r.t., it was quenched with water (10 mL), and
extracted with ethyl acetate (10 mL.times.3). The combined organic
layers were washed with brine (3 mL), dried over anhydrous sodium
sulfate and concentrated under vacuum. The residue was purified by
silica gel column chromatography using ethyl acetate/petroleum
ether (1:1) as eluent to afford the desired product Di-tert-butyl
2, 2'-((piperazine-1,4-diylbis(propane-3,1-diyl))bis(oxy))diacetate
as a colorless oil (200 mg, 46.5% yield). .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 3.94 (s, 4H), 3.56 (t, J=6.4 Hz, 4H),
2.42-2.47 (m, 12H), 1.79-1.83 (m, 4H), 1.48 (s, 18H). Chemical
Formula: C.sub.22H.sub.42N.sub.2O.sub.6; Molecular Weight:
430.59
2. Step--Synthesis of
2-(3-(4-(3-(2-(Tert-butoxy)-2-oxoethoxy)propyl)piperazin-1-yl)propoxy)ace-
tic acid
##STR00677##
[1649] To a solution of Di-tert-butyl 2,
2'-((piperazine-1,4-diylbis(propane-3,1-diyl))bis(oxy))diacetate
(1.5 g, 4.01 mmol) in MeOH (30 mL) and water (10 mL) was added NaOH
(160 mg, 4.01 mmol) at r.t. The resulting mixture was stirred at rt
overnight. The pH was adjusted to 6 with 2N HCl, and the mixture
was extracted with DCM (30 ml.times.2). The combined organic layers
were dried over anhydrous sodium sulfate and concentrated under
vacuum to afford the crude compound
2-(3-(4-(3-(2-(Tert-butoxy)-2-oxoethoxy)propyl)piperazin-1-yl)propoxy)ace-
tic acid as a colorless oil (1.0 g), which was used in next step
without further purification. Chemical Formula:
C.sub.18H.sub.34N.sub.2O.sub.6; Molecular Weight: 374.48
3. Step--Synthesis of Tert-butyl
2-(3-(4-(3-(2-((4-(isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimid-
in-4-yl)amino)pyridin-3-yl)amino)-2-oxoethoxy)propyl)piperazin-1-yl)propox-
y)acetate
##STR00678##
[1651] To a mixture of
2-(3-(4-(3-(2-(Tert-butoxy)-2-oxoethoxy)propyl)piperazin-1-yl)propoxy)ace-
tic acid (53 mg, 0.14 mmol),
N.sup.4-Isopropyl-N.sup.2-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)pyri-
dine-2,4,5-triamine (50 mg, 0.14 mmol), and DIPEA (72 mg, 0.56
mmol) in DMF (5 mL) was added HATU (106 mg, 0.28 mmol). The
reaction mixture was stirred at rt for 1 h. Then the reaction
mixture was diluted with 10 mL water and the resulting reaction
mixture was extracted with DCM (2.times.30 mL). The combined
organic layers were washed with brine (10 mL), dried over
Na.sub.2SO.sub.4 and filtered. The solvent was evaporated under
reduced pressure. The residue was purified by column chromatography
(DCM:MeOH=20:1) to afford the crude desired compound Tert-butyl
2-(3-(4-(3-(2-((4-(isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimid-
in-4-yl)amino)pyridin-3-yl)amino)-2-oxoethoxy)propyl)piperazin-1-yl)propox-
y)acetate as a colorless oil (50 mg, 0.07 mmol, 50.1%). LC-MS:
(ES.sup.+): m/z 714.3 [M+H]. t.sub.R=2.98 min Chemical Formula:
C.sub.36H.sub.59N.sub.9O.sub.6; Molecular Weight: 713.92
4. Step--Synthesis of
2-(3-(4-(3-((1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)am-
ino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)propyl)piperazin-1-yl)propoxy)a-
cetic acid
##STR00679##
[1653] A solution of Tert-butyl
2-(3-(4-(3-(2-((4-(isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimid-
in-4-yl)amino)pyridin-3-yl)amino)-2-oxoethoxy)propyl)piperazin-1-yl)propox-
y)acetate (200 mg, 0.28 mmol) in HOAc (5 mL) was heated to
150.degree. C. under microwave for 6 h. Then it was concentrated in
vacuo to afford the crude desired product
2-(3-(4-(3-((1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)am-
ino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)propyl)piperazin-1-yl)propoxy)a-
cetic acid (200 mg, crude). LC-MS: (ES.sup.+): m/z 640.3 [M+H].
t.sub.R=2.37 min Chemical Formula: C.sub.32H.sub.49N.sub.9O.sub.5;
Molecular Weight: 639.80
5. Step--Synthesis of
(2S,4R)-4-hydroxy-1-((S)-2-(2-(3-(4-(3-((1-Isopropyl-6-((2-(4-methoxypipe-
ridin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)pro-
pyl)piperazin-1-yl)propoxy)acetamido)-3,3-dimethylbutanoyl)-N--((S)-1-(4-(-
4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00680##
[1655] To a mixture of
2-(3-(4-(3-((1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)am-
ino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)propyl)piperazin-1-yl)propoxy)a-
cetic acid (100 mg, crude),
(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N--((S)-1-(4-(4-me-
thylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (45 mg,
0.10 mmol), and DIPEA (48 mg, 0.37 mmol) in DCM (5 mL) was added
HATU (80 mg, 0.21 mmol). The reaction mixture was stirred at rt for
1 h. Then the reaction mixture was diluted with water (10 mL), and
the resulting reaction mixture was extracted with DCM (30
mL.times.2). The combined organic layers were washed with brine (10
mL), dried over Na.sub.2SO.sub.4 and filtered. The solvent was
evaporated under reduced pressure. The residue was purified by
prep-TLC (DCM:MeOH=20:1) to afford the desired compound
(2S,4R)-4-hydroxy-1-((S)-2-(2-(3-(4-(3-((1-Isopropyl-6-((2-(4-methoxypipe-
ridin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methoxy)pro-
pyl)piperazin-1-yl)propoxy)acetamido)-3,3-dimethylbutanoyl)-N--((S)-1-(4-(-
4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (10 mg,
0.0094 mmol, 9.4%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 8.67 (d, J=4.4 Hz, 2H), 8.45 (s, 1H), 8.04 (d,
J=5.6 Hz, 1H), 7.46 (d, J=7.6 Hz, 1H), 7.37-7.42 (m, 6H), 7.18 (d,
J=8.4 Hz, 1H), 6.04 (d, J=5.2 Hz, 1H), 5.34-5.35 (m, 1H), 5.02-5.12
(m, 1H), 4.85-4.95 (m, 1H), 4.75 (s, 3H), 4.51-4.53 (m, 2H),
4.36-4.39 (m, 2H), 4.12-4.15 (m, 1H), 3.93-3.95 (m, 2H), 3.46-3.56
(m, 8H), 3.42 (s, 3H), 2.53 (s, 3H), 2.41-2.45 (m, 12H), 2.20-2.22
(m, 1H), 1.65-1.67 (m, 5H), 1.64 (s, 9H), 1.47 (d, J=4.8 Hz, 3H),
1.06 (s, 9H). LC-MS: (ES.sup.+): m/z 1066.6 [M+H]. t.sub.R=2.88 min
Chemical Formula: C.sub.55H.sub.79N.sub.13O.sub.7S; Molecular
Weight: 1066.38
Synthesis of Examples 97, 98, 99, and 100
##STR00681##
[1656]
4-(4-((3-Chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)quinazolin-6-yl-
)phenol
[1657] A suspension of
N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6-iodo-quinazolin-4-amine
(300 mg, 0.59 mmol) in a mixture of 1,2-Dimethoxyethane (12 ml) and
Ethanol (8 ml) was evacuated in vacuum and purged with argon
(5.times.), then 2M Na.sub.2CO.sub.3 in water (6.5 ml) was added
and the reaction mixture was again evacuated in vacuum and purged
with argon (5.times.), then
[4-[tert-butyl(dimethyl)silyl]oxyphenyl]boronic acid (209 mg, 0.831
mmol) was added into, and [(Ph).sub.3P].sub.2PdCl.sub.2 (70 mg,
0.08 mmol). The reaction mixture was heated to 60.degree. C. for 3
h. The reaction mixture was cooled to room temperature and the
reaction mixture was poured into an aqueous saturated solution of
NaHCO.sub.3(30 mL) and product was extracted with AcOEt (2.times.30
mL). Organic extracts were combined, dried (Na.sub.2SO.sub.4),
filtered over a celite pad, and evaporated under vacuum. Crude
product was purified by flash chromatography (SiO.sub.2-25 g. dry
silica-dispersion loading, gradient Hex:AcOEt, 9:1 to 100% AcOEt in
15 min) to give 210 mg of product
4-(4-((3-Chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)quinazolin-6-yl)pheno-
l (75% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.85 (s, 1H),
9.68 (s, 1H), 8.69 (s, 1H), 8.56 (s, 1H), 8.12 (d, J=8.7 Hz, 1H),
8.03 (s, 1H), 7.87-7.67 (m, 4H), 7.48 (td, J=8.0, 6.0 Hz, 1H),
7.39-7.25 (m, 3H), 7.19 (tt, J=7.8, 1.4 Hz, 1H), 6.93 (d, J=8.6 Hz,
2H), 5.27 (s, 2H). .sup.13C NMR (151 MHz, dmso) .delta. 163.06,
161.44, 157.66, 157.59, 154.07, 149.69, 148.44, 139.73, 139.69,
138.23, 133.18, 131.45, 130.69, 130.64, 129.86, 128.36, 128.32,
124.20, 123.45, 123.43, 122.38, 121.01, 118.95, 115.88, 115.33,
114.87, 114.73, 114.28, 114.21, 114.07, 69.37. LC-MS (ESI): m/z
[M+H].sup.+ Calcd. for C.sub.27H.sub.20ClFN.sub.3O.sub.2, 472.1228.
Found 472.1283.
##STR00682##
tert-Butyl
2-(2-(2-(4-(4-((3-chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)quinazolin-6-
-yl)phenoxy)-ethoxy)ethoxy)acetate
[1658] To a mixture of
4-[4-[3-chloro-4-[(3-fluorophenyl)methoxy]anilino]-quinazolin-6-yl]phenol
(10.4 mg, 0.022 mmol) and tert-butyl
2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]-acetate (10.8 mg, 0.03
mmol) in N,N-Dimethylformamide (2 mL) was added Cs.sub.2CO.sub.3
(21.7 mg, 0.067 mmol). Reaction mixture was heated at 50.degree. C.
for 6 h. Reaction mixture was diluted with AcOEt (20 mL), washed
with water (4.times.15 mL), dried Na.sub.2SO.sub.4 and evaporated
under vacuum. Crude product was purified by PTLC
(DCM:MeOH:NH.sub.4OH, 92:7:1) to give 8 mg of pure product
tert-Butyl
2-(2-(2-(4-(4-((3-chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)quinazolin-6-
-yl)phenoxy)-ethoxy)ethoxy)acetate (53% yield). .sup.1H NMR (500
MHz, DMSO-d6) .delta. 9.88 (s, 1H), 8.74 (s, 1H), 8.57 (s, 1H),
8.16 (d, J=8.7 Hz, 1H), 8.03 (s, 1H), 7.83 (dd, J=8.7, 6.6 Hz, 3H),
7.76 (dd, J=8.9, 2.6 Hz, 1H), 7.52-7.40 (m, 1H), 7.38-7.25 (m, 3H),
7.21-7.15 (m, 1H), 7.13 (d, J=8.8 Hz, 2H), 5.26 (s, 2H), 4.26-4.12
(m, 2H), 4.01 (s, 2H), 3.83-3.71 (m, 2H), 3.63 (s, 4H), 1.42 (s,
9H). .sup.13C NMR (151 MHz, dmso) .delta. 169.37, 163.01, 161.39,
158.54, 157.60, 154.15, 149.70, 148.58, 139.69, 139.64, 137.71,
133.15, 131.46, 131.44, 130.60, 130.55, 128.33, 128.25, 124.19,
123.36, 123.34, 122.36, 121.03, 119.29, 115.29, 115.00, 114.78,
114.64, 114.30, 114.12, 113.98, 80.66, 69.88, 69.40, 69.38, 68.91,
68.12, 67.28, 27.77. LC-MS (ESI): m/z [M+H].sup.+ Calcd. for
C.sub.37H.sub.38ClFN.sub.3O.sub.6, 674.2433. Found 674.2411.
tert-Butyl
2-(2-(2-(2-(4-(4-((3-chloro-4-((3-fluorobenzyl)oxy)phenyl)amino-
)quinazolin-6-yl)phenoxy)-ethoxy)ethoxy)ethoxy)acetate
[1659] To a mixture of
4-[4-[3-chloro-4-[(3-fluorophenyl)methoxy]anilino]-quinazolin-6-yl]phenol
(7.2 mg, 0.015 mmol) and tert-butyl
2-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]-ethoxy]-acetate (8.3
mg, 0.02 mmol) in N,N-Dimethylformamide (2 mL) was added
Cs.sub.2CO.sub.3 (14.91 mg, 0.05 mmol). Reaction mixture was heated
at 50.degree. C. for 2 h. Reaction mixture was diluted with AcOEt
(20 mL), washed with water (4.times.15 mL), dried Na.sub.2SO4 and
evaporated under vacuum. Crude product was purified by PTLC
(DCM:MeOH:NH.sub.4OH, 92:7:1) to give 10 mg of product tert-Butyl
2-(2-(2-(2-(4-(4-((3-chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)quinazoli-
n-6-yl)phenoxy)-ethoxy)ethoxy)ethoxy)acetate (91% yield). .sup.1H
NMR (500 MHz, DMSO-d6) .delta. 9.87 (s, 1H), 8.74 (d, J=2.1 Hz,
1H), 8.57 (s, 1H), 8.16 (dd, J=8.8, 1.9 Hz, 1H), 8.03 (s, 1H), 7.83
(dd, J=8.6, 5.7 Hz, 3H), 7.76 (dd, J=9.0, 2.6 Hz, 1H), 7.47 (td,
J=8.0, 6.1 Hz, 1H), 7.38-7.25 (m, 3H), 7.22-7.14 (m, 1H), 7.13 (d,
J=8.8 Hz, 2H), 5.27 (s, 2H), 4.23-4.13 (m, 2H), 3.98 (s, 2H),
3.84-3.74 (m, 2H), 3.68-3.46 (m, 8H), 1.41 (s, 9H). .sup.13C NMR
(151 MHz, DMSO-d6) .delta. 169.36, 163.01, 161.39, 158.54, 157.60,
154.15, 149.70, 148.57, 139.69, 139.64, 137.72, 133.14, 131.44,
130.61, 130.55, 128.32, 128.25, 124.19, 123.36, 123.34, 122.37,
121.03, 119.29, 115.28, 115.02, 114.78, 114.64, 114.30, 114.13,
113.98, 80.64, 69.95, 69.86, 69.78, 69.72, 69.38, 68.95, 68.09,
67.29, 27.76. LC-MS (ESI): m/z [M+H].sup.+ Calcd. For
C.sub.39H.sub.42ClFN.sub.3O.sub.7, 718.2695. Found 718.3026.
##STR00683##
Synthesis of Example 97 and 98
##STR00684##
[1660]
(2S,4R)-1-((S)-2-(2-(2-(2-(4-(4-((3-Chloro-4-((3-fluorobenzyl)oxy)p-
henyl)amino)quinazolin-6-yl)phenoxy)ethoxy)ethoxy)acetamido)-3,3-dimethylb-
utanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxa-
mide (Example 97)
[1661] A solution of tert-butyl
2-[2-[2-[4-[4-[3-chloro-4-[(3-fluorophenyl)methoxy]anilino]quinazolin-6-y-
l]-phenoxy]ethoxy]ethoxy]acetate (8 mg, 0.01 mmol) in a mixture of
TFA (1 ml, 13.46 mmol) and Dichloromethane (3 ml) was stirred for 2
h. Then the solvent was removed under vacuum and crude product was
dried under high vacuum for 2 h. The crude product was used in the
next step without any further purification (7.3 mg, quantitative
yield). LC-MS (ESI): m/z [M+H].sup.+ Calcd. for
C.sub.33H.sub.30ClFN.sub.3O.sub.6, 618.1807. Found 618.1917.
[1662] To a solution of
2-[2-[2-[4-[4-[3-chloro-4-[(3-fluorophenyl)methoxy]anilino]quinazolin-6-y-
l]phenoxy]-ethoxy]ethoxy]acetic acid (7.3 mg, 0.01 mmol), and
(2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[4-(4-methylt-
hiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide hydrochloride
(8.27 mg, 0.02 mmol) in N,N-Dimethylformamide (2 ml) was added
DIPEA (0.2 ml, 1.14 mmol) and HATU (8.98 mg, 0.02 mmol) at room
temperature. The reaction mixture was stirred for 12 h (overnight)
at the same temperature. Reaction mixture was diluted with AcOEt
(20 mL), washed with water (4.times.15 mL), dried
(Na.sub.2SO.sub.4) and evaporated under vacuum. Crude product was
purified by PTLC (DCM:MeOH:NH.sub.4OH, 92:7:1), to give 12 mg of
the expected product
(2S,4R)-1-((S)-2-(2-(2-(2-(4-(4-((3-Chloro-4-((3-fluorobenzyl)oxy)phenyl)-
amino)quinazolin-6-yl)phenoxy)ethoxy)ethoxy)acetamido)-3,3-dimethylbutanoy-
l)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
(98% yield). .sup.1H NMR (500 MHz, DMSO-d6) .delta. 9.87 (s, 1H),
8.95 (s, 1H), 8.72 (s, 1H), 8.57 (d, J=2.6 Hz, 2H), 8.13 (dd,
J=8.7, 1.9 Hz, 1H), 8.02 (d, J=2.4 Hz, 1H), 7.81 (dd, J=8.6, 4.9
Hz, 3H), 7.76 (dd, J=9.0, 2.6 Hz, 1H), 7.55-7.24 (m, 9H), 7.18 (t,
J=8.6 Hz, 1H), 7.10 (d, J=8.7 Hz, 2H), 5.26 (s, 2H), 5.15 (d, J=3.4
Hz, 1H), 4.58 (d, J=9.6 Hz, 1H), 4.50-4.22 (m, 5H), 4.19 (t, J=4.6
Hz, 2H), 4.00 (s, 2H), 3.89-3.77 (m, 2H), 3.75-3.54 (m, 6H), 2.41
(s, 3H), 2.11-2.01 (m, 1H), 1.95-1.87 (m, 1H), 0.95 (s, 9H).
.sup.13C NMR (151 MHz, DMSO-d6) .delta. 171.75, 169.16, 168.62,
163.01, 161.40, 158.51, 157.60, 154.15, 151.43, 149.70, 148.58,
147.72, 139.69, 139.64, 139.40, 137.73, 133.15, 131.43, 131.12,
130.61, 130.56, 129.69, 128.67, 128.32, 128.24, 128.15, 127.43,
124.18, 123.37, 123.35, 122.35, 121.04, 119.28, 115.29, 114.99,
114.79, 114.65, 114.30, 114.13, 113.99, 70.48, 69.79, 69.63, 69.40,
69.02, 68.89, 67.21, 58.76, 56.62, 55.73, 41.69, 37.96, 35.74,
26.21, 15.92. LC-MS (ESI): m/z [M+H].sup.+ Calcd. for
C.sub.55H.sub.58ClFN.sub.7O.sub.8S, 1030.3740. Found 1030.4004.
(2R,4S)-1-((S)-2-(2-(2-(2-(4-(4-((3-Chloro-4-((3-fluorobenzyl)oxy)phenyl)a-
mino)quinazolin-6-yl)phenoxy)ethoxy)ethoxy)acetamido)-3,3-dimethylbutanoyl-
)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
(Example 98)
[1663] It was prepared from
(2R,4S)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthia-
zol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride and
following the same procedure as describe above for example 97.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.67 (s, 1H), 8.62 (s,
1H), 8.54 (s, 1H), 7.96 (dd, J=8.7, 1.7 Hz, 1H), 7.91 (d, J=8.6 Hz,
1H), 7.84 (dd, J=8.8, 2.6 Hz, 1H), 7.71 (d, J=2.5 Hz, 1H), 7.56 (t,
J=6.8 Hz, 3H), 7.34 (dd, J=8.0, 6.2 Hz, 2H), 7.22 (t, J=9.3 Hz,
2H), 7.14 (t, J=8.3 Hz, 4H), 7.07-6.99 (m, 3H), 6.96 (d, J=8.9 Hz,
1H), 5.14 (s, 2H), 4.88 (dd, J=8.7, 5.0 Hz, 1H), 4.62 (p, J=5.4 Hz,
1H), 4.36 (dt, J=15.4, 7.3 Hz, 2H), 4.29 (d, J=6.6 Hz, 1H),
4.17-4.06 (m, 5H), 3.91-3.81 (m, 3H), 3.73-3.58 (m, 4H), 3.47-3.36
(m, 2H), 2.41 (s, 3H), 2.38 (t, J=5.3 Hz, 1H), 2.27 (ddd, J=13.8,
8.7, 5.9 Hz, 1H), 1.12 (s, 9H). .sup.13C NMR (151 MHz, CDCl.sub.3)
.delta. 171.85, 171.65, 170.60, 163.79, 162.16, 158.70, 158.07,
154.36, 150.50, 150.20, 148.29, 139.19, 139.14, 139.01, 137.91,
133.24, 132.47, 131.70, 131.65, 130.41, 130.18, 130.12, 129.14,
128.38, 127.54, 124.48, 122.92, 122.45, 122.44, 122.00, 119.24,
115.69, 115.55, 114.94, 114.80, 114.24, 114.04, 113.89, 71.39,
70.69, 70.44, 70.43, 70.17, 69.68, 67.94, 59.67, 58.55, 54.89,
42.89, 38.16, 33.85, 26.66, 16.10. LC-MS (ESI): m/z [M+H].sup.+:
Calcd. for C.sub.55H.sub.58ClFN.sub.7O.sub.8S, 1030.3740. Found
1030.3821.
##STR00685##
Synthesis of Example 99 and 100
##STR00686##
[1664]
(2S,4R)-1-((S)-2-(tert-Butyl)-14-(4-(4-((3-chloro-4-((3-fluorobenzy-
l)oxy)phenyl)amino)quinazolin-6-yl)phenoxy)-4-oxo-6,9,12-trioxa-3-azatetra-
decanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carbox-
amide
[1665] It was prepared from
(2R,4S)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthia-
zol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (8) and
tert-butyl
2-(2-(2-(2-(4-(4-((3-chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)-quinazol-
in-6-yl)phenoxy)ethoxy)ethoxy)ethoxy)acetate, following the same
procedure as described above for example 97 (93% yield). .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 9.87 (s, 1H), 8.95 (s, 1H), 8.72 (s,
1H), 8.59 (t, J=6.0 Hz, 1H), 8.55 (s, 1H), 8.13 (dd, J=8.7, 1.9 Hz,
1H), 8.00 (d, J=2.5 Hz, 1H), 7.89-7.76 (m, 3H), 7.73 (dd, J=9.0,
2.6 Hz, 1H), 7.54-7.23 (m, 8H), 7.22-7.12 (m, 1H), 7.09 (d, J=8.8
Hz, 2H), 5.24 (s, 2H), 5.15 (d, J=3.5 Hz, 1H), 4.55 (d, J=9.6 Hz,
1H), 4.48-4.18 (m, 5H), 4.18-4.06 (m, 2H), 3.95 (s, 2H), 3.80-3.69
(m, 2H), 3.69-3.51 (m, 8H), 2.41 (s, 3H), 2.08-2.00 (m, 1H),
1.93-1.82 (m, 1H), 0.92 (s, 9H). .sup.13C NMR (151 MHz, DMSO-d6)
.delta. 171.76, 169.12, 168.59, 163.01, 161.39, 158.53, 157.60,
154.15, 151.45, 149.70, 148.57, 147.73, 139.68, 139.63, 139.42,
137.72, 133.14, 131.44, 131.42, 131.13, 130.60, 130.55, 129.68,
128.68, 128.32, 128.24, 127.44, 124.19, 123.36, 123.34, 122.36,
121.03, 119.27, 115.28, 114.99, 114.78, 114.64, 114.29, 114.13,
113.98, 70.48, 69.97, 69.90, 69.63, 69.59, 69.38, 68.95, 68.88,
67.26, 58.75, 56.60, 55.69, 41.68, 37.94, 35.73, 26.19, 15.93.
LC-MS (ESI): m/z [M+H].sup.+: Calcd. for
C.sub.57H.sub.62ClFN.sub.7O.sub.9S, 1074.4002. Found 1074.4285.
(2S,4S)-1-((S)-2-(tert-Butyl)-14-(4-(4-((3-chloro-4-((3-fluorobenzyl)oxy)p-
henyl)amino)quinazolin-6-yl)phenoxy)-4-oxo-6,9,12-trioxa-3-azatetradecanoy-
l)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
(example 100).)
[1666] It was prepared from
(2S,4S)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthia-
zol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (10) and
tert-butyl
2-(2-(2-(2-(4-(4-((3-chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)-quinazol-
in-6-yl)phenoxy)ethoxy)ethoxy)ethoxy)acetate, following the same
procedure as reported above for example 97 (63% yield). .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 9.95 (s, 1H), 9.04 (s, 1H), 8.84-8.77
(m, 1H), 8.73 (t, J=6.0 Hz, 1H), 8.64 (s, 1H), 8.22 (dd, J=8.8, 1.8
Hz, 1H), 8.10 (d, J=2.6 Hz, 1H), 7.89 (dd, J=8.8, 1.8 Hz, 3H), 7.83
(dd, J=9.0, 2.6 Hz, 1H), 7.65-7.30 (m, 9H), 7.25 (td, J=8.8, 8.3,
2.6 Hz, 1H), 7.18 (d, J=8.8 Hz, 2H), 5.51 (d, J=7.2 Hz, 1H), 5.33
(s, 2H), 4.59 (d, J=9.2 Hz, 1H), 4.52-4.41 (m, 2H), 4.40-4.25 (m,
2H), 4.26-4.19 (m, 2H), 4.03 (s, 2H), 3.99-3.91 (m, 1H), 3.88-3.80
(m, 2H), 3.68 (ddt, J=6.7, 5.2, 3.3 Hz, 8H), 3.59-3.44 (m, 1H),
2.50 (s, 3H), 2.44-2.35 (m, 1H), 1.81 (dt, J=12.4, 6.1 Hz, 1H),
1.03 (s, 9H). .sup.13C NMR (151 MHz, dmso) .delta. 172.26, 169.38,
168.91, 163.02, 161.40, 158.53, 157.61, 154.16, 151.48, 149.71,
148.58, 147.76, 139.69, 139.64, 139.14, 137.72, 133.15, 131.45,
131.11, 130.61, 130.56, 129.77, 128.70, 128.34, 128.25, 127.47,
124.20, 123.37, 123.35, 122.38, 121.04, 119.29, 115.29, 115.00,
114.79, 114.65, 114.31, 114.13, 113.99, 70.46, 69.96, 69.88, 69.64,
69.52, 69.40, 69.03, 68.97, 67.27, 58.59, 55.84, 55.62, 41.82,
36.92, 35.19, 26.18, 15.94. LC-MS (ESI): m/z [M+H].sup.+: Calcd.
for C.sub.57H.sub.62ClFN.sub.7O.sub.9S, 1074.4002. Found
1074.3920.
Synthesis of Example 102
5-(2-(2-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)b-
enzyl)piperazin-1-yl)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-
-1,3-dione
##STR00687##
[1668] Synthetic Scheme:
##STR00688##
1. Step--Synthesis of
2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)ben-
zyl)piperazin-1-yl)ethoxy)ethan-1-ol
##STR00689##
[1670] A solution of
N-(2-chlorobenzyl)-6-(4-(piperazin-1-ylmethyl)phenyl)-7H-pyrrolo[2,3-d]py-
rimidin-4-amine (300 mg, 0.693 mmol), 2-(2-hydroxyethoxy)ethyl
4-methylbenzenesulfonate (900 mg, 3.464 mmol), and K.sub.2CO.sub.3
(478 mg, 3.464 mmol) in DMF (8 mL) was stirred at 75.degree. C.
overnight. After cooling to rt, the reaction was quenched with
water, and the mixture was extracted with ethyl acetate (20
mL.times.3). The combined organic layers were dried over anhydrous
sodium sulfate and concentrated under vacuum to afford crude title
compound
2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)ben-
zyl)piperazin-1-yl)ethoxy)ethan-1-ol (250 mg), which was used to
next step without further purification.
2. Step--Synthesis of
5-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
benzyl)piperazin-1-yl)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin-
e-1,3-dione
##STR00690##
[1672] To a solution of
2-(2-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)ben-
zyl)piperazin-1-yl)ethoxy)ethan-1-ol (50 mg, crude, 0.096 mmol),
PPh.sub.3 (126 mg, 0.480 mmol) and
2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (26 mg,
0.096 mmol) in dry THF (10 mL) was added DIAD (97 mg, 0.480 mmol)
dropwise at 0.degree. C. under N.sub.2. The mixture was stirred at
room temperature for 1 h. The reaction mixture was quenched with
water (50 mL), and the mixture was taken up with EA. The combined
organic layers were washed with brine, dried over MgSO.sub.4, and
concentrated. The residue was purified by chromatography (silica
gel, DCM: MeOH (10:1, v:v)) to afford the title compound
5-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
benzyl)piperazin-1-yl)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin-
e-1,3-dione (6 mg, 0.008 mmol). .sup.1H NMR (400 MHz, DMSO-d6):
.delta. 12.09 (s, 1H), 8.18 (s, 1H), 8.10 (s, 1H), 8.05 (s, 1H),
7.74 (d, J=7.2 Hz, 3H), 7.46 (d, J=3.6 Hz, 1H), 7.35 (d, J=8.0 Hz,
3H), 7.28-7.30 (m, 2H), 7.15-7.18 (m, 2H), 7.00 (s, 1H), 5.13-5.18
(m, 1H), 4.78 (d, J=5.2 Hz, 2H), 3.86-3.89 (m, 2H), 3.75-3.79 (m,
2H), 2.72-3.04 (m, 3H), 2.35-2.48 (m, 9H), 2.08-2.13 (m, 1H).
Synthesis of Example 106
(2S,4R)--N-(2-(2-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimid-
in-6-yl)benzyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hy-
droxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carbo-
xamide
##STR00691##
[1674] Synthetic Scheme:
##STR00692##
1. Step--Synthesis of
(2S,4R)--N-(2-(2,2-Diethoxyethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydr-
oxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxa-
mide
##STR00693##
[1676] To a solution of
(2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-3-m-
ethyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamide
(100 mg, 0.18 mmol) in DMF (5 mL) was added K.sub.2CO.sub.3 (75.6
mg, 0.55 mmol) and 2-bromo-1,1-diethoxyethane (53.9 mg, 0.27 mmol)
at 25.degree. C. The resulting solution was stirred at 110.degree.
C. for 16 h. After cooling to rt, the reaction was quenched with
H.sub.2O (10 mL), and the mixture was extracted with EtOAc (10
mL.times.2). The combined organic layers were dried over anhydrous
sodium sulfate and concentration to give the title product
(2S,4R)--N-(2-(2,2-Diethoxyethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydr-
oxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxa-
mide (90 mg, crude), which was used in the next step without
further purification.
2. Step--Synthesis of
(2S,4R)-4-Hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)-N-(4--
(4-methylthiazol-5-yl)-2-(2-oxoethoxy)benzyl)pyrrolidine-2-carboxamide
##STR00694##
[1678] To a solution
(2S,4R)--N-(2-(2,2-Diethoxyethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydr-
oxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxa-
mide (90 mg, 0.14 mmol) in CH.sub.3CN/H.sub.2O (5 mL/2 mL) was
added 1 N HCl (2 mL) at 25.degree. C. The reaction was stirred at
80.degree. C. for 2 h. After cooling to 25.degree. C., the reaction
was quenched with NaHCO.sub.3(10 mL), and the mixture was extracted
with EtOAc (10 mL.times.2). The combined organic layers were washed
with brine (10 mL.times.2), dried over anhydrous sodium sulfate and
concentration to afford the title product
(2S,4R)-4-Hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)-N-(4--
(4-methylthiazol-5-yl)-2-(2-oxoethoxy)benzyl)pyrrolidine-2-carboxamide
(90 mg, crude), which was used in the next step without further
purification.
3. Step--Synthesis of
(2S,4R)--N-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimi-
din-6-yl)benzyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-h-
ydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carb-
oxamide
##STR00695##
[1680] To a solution of
(2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)-N-(4--
(4-methylthiazol-5-yl)-2-(2-oxoethoxy)benzyl)pyrrolidine-2-carboxamide
(90 mg, 0.15 mmol) and
N-(2-chlorobenzyl)-6-(4-(piperazin-1-ylmethyl)phenyl)-7H-pyrrolo[2,3-d]py-
rimidin-4-amine (79.2 mg, 0.18 mmol) in DMSO/MeOH (2 mL/2 mL) was
added NaBH.sub.3CN (47.9 mg, 0.76 mmol) at 10.degree. C. The
resulting mixture was stirred at 40.degree. C. for 3 h. After
cooling to 20.degree. C., the reaction was quenched with H.sub.2O
(10 mL), and the mixture was extracted with EtOAc (10 mL.times.2).
The combined organic layers were washed with brine (10 mL.times.2),
dried over anhydrous sodium sulfate and concentrated. The residue
was purified with silica gel column and prep-HPLC to afford the
title product
(2S,4R)--N-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimi-
din-6-yl)benzyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-h-
ydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carb-
oxamide (3 mg: 99% purity and 6.5 mg: 86% purity, 3 steps 7.6%).
.sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 8.88 (s, 1H), 8.09 (s,
1H), 7.73-7.76 (m, 3H), 7.37-7.57 (m, 9H), 7.22-7.24 (m, 2H),
7.03-7.05 (m, 2H), 6.93 (s, 1H), 4.95 (s, 2H), 4.39-4.60 (m, 7H),
4.24 (s, 2H), 3.82-3.96 (m, 2H), 3.57 (s, 1H), 2.96 (s, 2H),
2.60-2.77 (m, 8H), 2.49 (s, 3H), 2.01-2.33 (m, 3H), 0.98 (d, J=6.4
Hz, 3H), 0.78 (d, J=6.4 Hz, 3H). Chemical Formula:
C.sub.55H.sub.59ClN.sub.10O.sub.5S; Molecular Weight: 1007.64 LCMS:
m/e+=504.3 [M+2H]2+; t.sub.R=3.33 min
Synthesis of Example 108
##STR00696##
[1682] Synthetic Route:
##STR00697##
1. Step--Synthesis of
2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
benzyl)piperazin-1-yl)ethoxy)ethoxy)ethan-1-ol
##STR00698##
[1684] A solution of
N-(2-chlorobenzyl)-6-(4-(piperazin-1-ylmethyl)phenyl)-7H-pyrrolo[2,3-d]py-
rimidin-4-amine (300 mg, 0.693 mmol),
2-(2-(2-hydroxyethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (900
mg, 3.464 mmol), and K.sub.2CO.sub.3 (478 mg, 3.464 mmol) in dry
DMF (8 mL) was stirred at 80.degree. C. overnight. After cooling to
room temperature, the reaction was quenched with water (20 mL), and
the mixture was extracted with ethyl acetate (20 mL.times.3). The
combined organic layers were dried over anhydrous Na.sub.2SO.sub.4
and concentrated to give title product
2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
benzyl)piperazin-1-yl)ethoxy)ethoxy)ethan-1-ol (300 mg, crude) as
light yellow solid, which was used to next step without further
purification.
2. Step--Synthesis of
2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
benzyl)piperazin-1-yl)ethoxy)ethoxy)ethyl methanesulfonate
##STR00699##
[1686] To a solution of
2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
benzyl)piperazin-1-yl)ethoxy)ethoxy)ethan-1-ol (300 mg, crude,
0.531 mmol) in DCM (10 mL) were added TEA (265 mg, 2.653 mmol) and
MsCl (182 mg, 1.593 mmol) at room temperature. After stirring for
20 min, the reaction was quenched with water, and the mixture was
extracted with DCM (30 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4 and concentrated to give
the title product
2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
benzyl)piperazin-1-yl)ethoxy)ethoxy)ethyl methanesulfonate (350 mg,
crude) as faint yellow solid, which was used in the next step
without further purification.
2. Step--Synthesis of
(2S,4R)--N-(2-(2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]-
pyrimidin-6-yl)benzyl)piperazin-1-yl)ethoxy)ethoxy)ethoxy)-4-(4-methylthia-
zol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoy-
l)pyrrolidine-2-carboxamide
##STR00700##
[1688] To a solution of
2-(2-(2-(4-(4-(4-((2-chlorobenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
benzyl)piperazin-1-yl)ethoxy)ethoxy)ethyl methanesulfonate (250 mg,
crude, 0.39 mmol) in dry DMF (5 ml) were added K.sub.2CO.sub.3 (108
mg, 0.78 mmol) and
(2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)--
1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamide
(215 mg, 0.39 mmol). The resulting solution was stirred at
75.degree. C. overnight. After cooling to rt, the reaction mixture
was quenched with water, and the mixture was extracted with EA (30
mL.times.2). The combined organic layers were dried over anhydrous
sodium sulfate and concentrated under vacuum. The residue was
purified by column chromatography to afford the title product
(2S,4R)--N-(2-(2-(2-(2-(4-(4-(4-((2-Chlorobenzyl)amino)-7H-pyrrolo[2,3-d]-
pyrimidin-6-yl)benzyl)piperazin-1-yl)ethoxy)ethoxy)ethoxy)-4-(4-methylthia-
zol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoy-
l)pyrrolidine-2-carboxamide (17 mg). 1H NMR (400 MHz, CDCl.sub.3):
.delta. 8.64 (s, 1H), 8.31 (s, 1H), 7.67 (d, J=7.2 Hz, 3H),
7.46-7.47 (m, 1H), 7.30-7.41 (m, 8H), 7.20 (s, 2H), 6.94 (d, J=5.6
Hz, 1H), 6.84 (s, 1H), 6.63 (s, 1H), 6.04 (s, 1H), 4.95 (d, J=4.4
Hz, 2H), 4.14-4.79 (m, 10H), 3.46-3.85 (m, 13H), 2.48-2.65 (m,
16H), 1.99 (d, J=6.0 Hz, 3H), 0.82 (d, J=6.4 Hz, 3H).
Synthesis of Examples 256 and 257
##STR00701##
[1689]
(2S,4R)--N-(2-(2-(2-((3-(2-((R)-1-(5-fluoro-2-hydroxyphenyl)-2-oxo--
2-(thiazol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)prop-2-yn-1-yl)oxy)ethoxy-
)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(1-ox-
oisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamide and
(2S,4R)--N-(2-(2-(2-((3-(2-((R)-1-(5-fluoro-2-hydroxyphenyl)-2-oxo-2-(thi-
azol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)prop-2-yn-1-yl)oxy)ethoxy)ethox-
y)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoin-
dolin-2-yl)butanoyl)pyrrolidine-2-carboxamide
##STR00702## ##STR00703##
[1690] Experimental Procedures
1. Step--Synthesis of
2-hydroxy-4-(4-methylthiazol-5-yl)benzonitrile
##STR00704##
[1692] Into a 500-ml-3-necked round-bottom flask with an inert
atmosphere of nitrogen were loaded 4-bromo-2-hydroxybenzonitrile
(26 g, 131.3 mmol, 1.00 equiv), DMA (300 ml), 4-methylthiazole (26
g, 262.6 mmol, 2.00 equiv), KOAc (26 g, 262.6 mmol, 2.00 equiv),
Pd(OAc)2 (884.3 mg, 3.94 mmol, 0.03 equiv). The resulting solution
was stirred for 5 hour at 150.degree. C. The reaction was then
quenched by the addition of 1000 mL of water. The resulting mixture
was washed with 3.times.500 mL of ethyl acetate and the organic
layers combined, and washed with 3.times.500 mL of H.sub.2O. The
mixture was dried over anhydrous sodium sulfate and concentrated
under vacuum. The residue was applied onto a silica gel column with
ethyl acetate/petroleum ether (1:1). This resulted in 14.4 g (66.66
mmol, 50.77%) of 2-hydroxy-4-(4-methylthiazol-5-yl) benzonitrile as
a yellow solid. .sup.1HNMR (400 MHz, DMSO-d6): .delta. 2.49 (s,
3H), 7.08 (d, J=8.0 Hz, 1H), 7.13 (s, 1H), 7.71 (d, J=8.0 Hz, 1H),
9.07 (s, 1H), 11.35 (s, 1H).
2. Step--Synthesis of
2-(aminomethyl)-5-(4-methylthiazol-5-yl)phenol
##STR00705##
[1694] Into a 1000-ml-3-necked round-bottom flash purged and
maintained with an inert atmosphere of nitrogen, was placed a
solution of 2-hydroxy-4-(4-methylthiazol-5-yl) benzonitrile (14.4
g, 66.66 mmol) in THF 400 ml. This was followed by the addition of
LiAlH.sub.4 (6.34 g, 166.67 mmol, 2.50 equiv) in several batches at
0.degree. C. The resulting mixture heated to reflux overnight, then
allowed to cool to room temperature. The mixture was filtered and
the filter cake was washed with 10% MeOH in DCM for four times. The
combined filtrates were concentrated to afford the crude product
2-(aminomethyl)-5-(4-methylthiazol-5-yl)phenol 10.4 g (47.27 mmol,
71% yield). It was used to next step without further purification.
.sup.1H NMR (400 MHz, DMSO-d6): .delta. 2.40 (s, 3H), 3.62 (br,
1H), 6.33 (d, J=6.0 Hz, 1H), 6.56 (s, 1H), 6.96 (d, J=7.6 Hz, 1H),
8.82 (s, 1H).
3. Step--Synthesis of (S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoic
acid
##STR00706##
[1696] (S)-2-amino-3-methylbutanoic acid (43.7 g, 373 mmol) was
added to a solution of phthalaldehyde (50 g, 373 mmol) in
acetonitrile (1000 mL). The resulting mixture was refluxed for
overnight. The reaction mixture was cooled to r.t then filtered and
dried to afford the desired compound
(S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoic acid (72 g, 83%).
4. Step--Synthesis of methyl
(2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrol-
idine-2-carboxylate
##STR00707##
[1698] A solution of (S)-3-methyl-2-(1-oxoisoindolin-2-yl) butanoic
acid (5 g, 21.44 mmol), (2S,4R)-methyl
4-hydroxypyrrolidine-2-carboxylate, HCl (4.67 g, 25.7 mmol) DIPEA
(8.98 ml, 51.4 mmol) in DMF (Volume: 30 ml) was added HATU (9.78 g,
25.7 mmol) at 0.degree. C., The resulting mixture was stirred at
room temperature for 2 hours. The mixture was partitioned between
EtOAc and water. The organic phase was washed with water, brine and
dried over anhydrous Na.sub.2SO.sub.4. The residue was purified
with column chromatography to afford the desired compound methyl
(2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrol-
idine-2-carboxylate (5.41 g, 70%). .sup.1H-NMR (400 MHz,
CDCl.sub.3): .delta. 0.84 (d, J=5.6 Hz, 3H), 1.09 (d, J=5.2 Hz,
3H), 2.00 (m, 1H), 2.31-2.41 (m, 2H), 3.76 (s, 3H), 3.84 (d, J=11.2
Hz, 1H), 4.30-4.38 (m, 2H), 4.56-4.71 (m, 3H), 4.78 (m, 1H),
7.27-7.42 (m, 3H), 7.69 (d, J=7.2 Hz, 1H).
5. Step--Synthesis of
(2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrol-
idine-2-carboxylic acid
##STR00708##
[1700] A solution of methyl (2S,4R)-methyl
4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl) butanoyl)
pyrrolidine-2-carboxylate (5 g, 13.87 mmol) in Water (Volume: 50
ml), THF (Volume: 100 ml), was added lithium hydroxide, H.sub.2O
(1.164 g, 27.7 mmol), at 0.degree. C. The reaction was stirred at
room temperature for 2 h. The reaction mixture was acidified with
1N HCl to pH1-2, and extracted with EtOAc. The combined organic
layer was washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated to afford the desired compound
(2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrol-
idine-2-carboxylic acid (4.42 g, 92%). .sup.1HNMR (400 MHz,
CDCl.sub.3): .delta. 0.87 (d, J=6.4 Hz, 3H), 1.05 (d, J=5.6 Hz,
3H), 2.21 (m, 1H), 2.31 (m, 1H), 2.43 (m, 1H), 3.80 (d, J=6.4 Hz,
1H), 4.37-4.44 (m, 2H), 4.55 (s, 1H), 4.64 (t, J=8.0 Hz, 7.6 Hz,
1H), 4.73 (d, J=17.6 Hz, 1H), 4.83 (d, J=10.8 Hz, 1H), 7.38-7.42
(m, 2H), 7.49 (d, J=7.2 Hz, 1H), 7.74 (d, J=7.6 Hz, 1H).
6. Step--Synthesis of
(2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-3-m-
ethyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamide
##STR00709##
[1702] To a solution of
2-(aminomethyl)-5-(4-methylthiazol-5-yl)phenol (6.00 g, 27.3 mmol,
1.10 equiv),
(2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrol-
idine-2-carboxylic acid (8.58 g, 24.79 mmol, 1.00equiv), EDCI (5.70
g, 29.75 mmol, 1.20equiv), HOBT (4.02 g, 29.75 mmol, 1.20equiv) in
CH.sub.2Cl.sub.2(100 mL), was added Et.sub.3N (6.0 g, 10.75 mmol).
The resulting solution was stirred at room temperature for 1 hour.
The mixture was partitioned between CH.sub.2Cl.sub.2 and water. The
organic phase was washed with water, brine and dried over anhydrous
Na.sub.2SO.sub.4. The residue was purified with column
chromatography to give the title compound
(2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-3-m-
ethyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamide
(6.3 g, 11.49 mmol, 46.3% yield) .sup.1HNMR (400 MHz, CDCl.sub.3):
.delta. 0.81 (d, J=6.4 Hz, 3H), 0.86 (d, J=6.8 Hz, 3H), 1.96-2.01
(m, 1H), 2.34-2.40 (m, 1H), 2.44-2.53 (m, 4H), 3.63 (dd, J=3.6,
12.0 Hz 1H), 4.27-4.2 (m, 1H), 4.38-4.43 (m, 2H), 4.53 (s, 2H),
4.68-4.71 (m, 3H), 6.91 (d, J=8.0 Hz, 1H), 7.01 (s, 1H), 7.13 (d,
J=7.6 Hz, 1H), 7.42-7.44 (m, 2H), 7.52 (d, J=7.2 Hz, 1H), 7.78 (d,
J=7.2 Hz, 1H), 8.01 (s, 1H), 8.66 (s, 1H), 9.20 (br, H). LC-MS
(ESI): calcd. 548.21; Found, 549.3 (M+H);
##STR00710## ##STR00711##
7. (Step--Synthesis of methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-hydroxyethoxy)ethoxy)prop--
1-yn-1-yl)-1-oxoisoindolin-2-yl)acetate
##STR00712##
[1704] To a stirred mixture of methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-iodo-1-oxoisoindolin-2-yl)acetate
(531 mg, 1.0 mmol), 2-(2-(prop-2-yn-1-yloxy)ethoxy)ethanol (288 mg,
2.0 mmol) and triethylamine (607 mg, 6.00 mmol) in acetonitrile (5
ml) were added copper(I) iodide (38 mg, 0.20 mmol) and
bis(triphenylphosphine)palladium(II) chloride (140 mg, 0.20 mmol)
under nitrogen atmosphere; the mixture was degassed with nitrogen
three times. The mixture was stirred at 65.degree. C. for 16 hours.
TLC showed the reaction was complete. The reaction mixture was
partitioned between ethyl acetate (50 ml) and water (20 ml). The
organic layer was collected, washed with brine (20 ml.times.2),
dried over anhydrous sodium sulfate and concentrated under reduced
pressure to give a crude residue which was purified by silica gel
flash column chromatography (eluted with 4% methanol in
dichloromethane) to afford methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-hydroxyethoxy)ethoxy)prop--
1-yn-1-yl)-1-oxoisoindolin-2-yl)acetate (450 mg, yield 82%) as
brown oil. LC_MS: (ES.sup.+): m/z 548.3 [M+H].sup.+. t.sub.R=2.848
min.
8. Step--Synthesis of methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(1-oxo-7-(3-(2-(2-(tosyloxy)ethoxy)eth-
oxy)prop-1-yn-1-yl)isoindolin-2-yl)acetate
##STR00713##
[1706] A mixture of methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-hydroxyethoxy)ethoxy)prop--
1-yn-1-yl)-1-oxoisoindolin-2-yl)acetate (450 mg, 0.82 mmol),
4-toluenesulfonyl chloride (235 mg, 1.23 mmol),
N,N-dimethylpyridin-4-amine (10 mg, 0.08 mmol), and triethylamine
(166 mg, 1.64 mmol) in dichloromethane (15 ml) was stirred at room
temperature overnight. TLC showed the reaction was complete. The
reaction mixture was diluted with dichloromethane (40 ml) and
washed with brine (40 ml.times.2), dried over anhydrous sodium
sulfate and concentrated under reduced pressure to give a crude
residue which was purified by silica gel flash column
chromatography (eluted with 4% methanol in dichloromethane) to
afford methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(1-oxo-7-(3-(2-(2-(tosyloxy)ethoxy)eth-
oxy)prop-1-yn-1-yl)isoindolin-2-yl)acetate (360 mg, yield 61%) as
white solid. LC_MS: (ES.sup.+): m/z 702.5 [M+H].sup.+.
t.sub.R=3.282 min.
9. Step--Synthesis of methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-(((2S,4R)-4-hydroxy-1-(-
(S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamido)me-
thyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-1-oxoi-
soindolin-2-yl)acetate
##STR00714##
[1708] A mixture of methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(1-oxo-7-(3-(2-(2-(tosyloxy)ethoxy)eth-
oxy)prop-1-yn-1-yl)isoindolin-2-yl)acetate (383 mg, 0.55 mmol),
(2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-3-m-
ethyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamide
(250 mg, 0.46 mmol) and potassium carbonate (127 mg, 0.92 mmol) in
N,N-dimethylformamide (8 ml) was stirred at 80.degree. C. under
nitrogen overnight. TLC showed the reaction was complete. The
reaction mixture was partitioned between ethyl acetate (50 ml) and
water (30 ml). The organic layer was collected, washed with brine
(20 ml.times.2), dried over anhydrous sodium sulfate and
concentrated under reduced pressure to give a crude residue which
was purified by silica gel flash column chromatography (eluted with
3% methanol in dichloromethane) to afford methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-(((2S,4R)-4-hydr-
oxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxa-
mido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-
-1-oxoisoindolin-2-yl)acetate (360 mg, yield 73%) as white solid.
LC_MS: (ES.sup.+): m/z 1078.4 [M+H].sup.+. t.sub.R=3.027 min.
10. Step--Synthesis of
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-(((2S,4R)-4-hydroxy-1-(-
(S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamido)me-
thyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-1-oxoi-
soindolin-2-yl)acetic acid
##STR00715##
[1710] A mixture of methyl
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-(((2S,4R)-4-hydroxy-1-(-
(S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamido)me-
thyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-1-oxoi-
soindolin-2-yl)acetate (360 mg, 0.33 mmol) and lithium hydroxide
monohydrate (28 mg, 0.67 mmol) in tetrahydrofuran (8 ml)-methanol
(2 ml)-water (2 ml) was stirred at room temperature for 16 hours.
TLC showed the reaction was complete. The reaction mixture was
acidified with diluted hydrochloride acid (1N) till pH 5-6, and
extracted with ethyl acetate (30 ml.times.2). The combined organic
layers were dried over sodium sulfate and concentrated under
reduced pressure to give
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-(((2S,4R)-4-hydroxy-1-(-
(S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamido)me-
thyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-1-oxoi-
soindolin-2-yl)acetic acid (400 mg, crude) as white solid which was
used in next step directly without further purification. LC_MS:
(ES.sup.+): m/z 1064.6 [M+H].sup.+. t.sub.R=2.882 min.
11. Step--Synthesis of
(2S,4R)--N-(2-(2-(2-((3-(2-(1-(2-(benzyloxy)-5-fluorophenyl)-2-oxo-2-(thi-
azol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)prop-2-yn-1-yl)oxy)ethoxy)ethox-
y)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoin-
dolin-2-yl)butanoyl)pyrrolidine-2-carboxamide
##STR00716##
[1712] To a stirred solution of
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-(((2S,4R)-4-hydroxy-1-(-
(S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamido)me-
thyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-1-oxoi-
soindolin-2-yl)acetic acid (400 mg, crude),
N-ethyl-N-isopropylpropan-2-amine (85 mg, 0.66 mmol) and
thiazol-2-amine (33 mg, 0.33 mmol) in N,N-dimethylformamide (5 ml)
was added
2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate (HATU) (250 mg, 0.66 mmol) at 0.degree. C., the
resulting mixture was allowed to warm to room temperature and
stirred for 20 min. TLC showed the reaction was complete. The
reaction mixture was partitioned between ethyl acetate (30 ml) and
water (20 ml). The organic layer was collected, washed with brine
(20 ml.times.2), dried over anhydrous sodium sulfate and
concentrated under reduced pressure to give a crude residue which
was purified by silica gel flash column chromatography (eluted with
5% methanol in dichloromethane) to afford
(2S,4R)--N-(2-(2-(2-((3-(2-(1-(2-(benzyloxy)-5-fluorophenyl)-2-oxo-2-(thi-
azol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)prop-2-yn-1-yl)oxy)ethoxy)ethox-
y)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoin-
dolin-2-yl)butanoyl)pyrrolidine-2-carboxamide (200 mg, yield 53%)
as white solid. LC_MS: (ES.sup.+): m/z 1146.5 [(M+1)/2+H].sup.+.
t.sub.R=3.010 min.
12. Step--Synthesis of
(2S,4R)--N-(2-(2-(2-((3-(2-(1-(5-fluoro-2-hydroxyphenyl)-2-oxo-2-(thiazol-
-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)prop-2-yn-1-yl)oxy)ethoxy)ethoxy)-4-
-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindoli-
n-2-yl)butanoyl)pyrrolidine-2-carboxamide
##STR00717##
[1714] To a stirred solution of
(2S,4R)--N-(2-(2-(2-((3-(2-(1-(2-(benzyloxy)-5-fluorophenyl)-2-oxo-2-(thi-
azol-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)prop-2-yn-1-yl)oxy)ethoxy)ethox-
y)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoin-
dolin-2-yl)butanoyl)pyrrolidine-2-carboxamide (100 mg, 0.09 mmol)
in dichloromethane (8 ml) was added boron tribromide (131 mg, 0.52
mmol) in dichloromethane (1 ml) at -70.degree. C. over 30 minutes.
The resulting mixture was stirred at -70.degree. C. for 1 hour. TLC
showed the reaction was complete. The reaction mixture was diluted
with dichloromethane (15 ml) and quenched with saturated sodium
bicarbonate solution (8 ml) at -70.degree. C. The organic layer was
collected and the aqueous layer was extracted with dichloromethane
(15 ml.times.2). The combined organic layers were washed with brine
(20 ml), dried over anhydrous sodium sulfate, and concentrated
under reduced pressure to give a crude residue which was purified
by preparative TLC (eluted with 5% methanol in dichloromethane) to
afford
(2S,4R)--N-(2-(2-(2-((3-(2-(1-(5-fluoro-2-hydroxyphenyl)-2-oxo-2-(thiazol-
-2-ylamino)ethyl)-3-oxoisoindolin-4-yl)prop-2-yn-1-yl)oxy)ethoxy)ethoxy)-4-
-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindoli-
n-2-yl)butanoyl)pyrrolidine-2-carboxamide (8.5 mg, yield 9%) as
white solid. LC_MS: (ES.sup.+): m/z 1056.5 [M+H].sup.+.
t.sub.R=2.790 min. .sup.1H NMR (400 MHz, DMSO-d6): .delta. 0.72 (d,
J=6.8 Hz, 3H), 0.95 (d, J=6.4 Hz, 3H), 1.88-1.95 (m, 1H), 2.01-2.07
(m, 1H), 2.29-2.35 (m, 1H), 2.46 (s, 3H), 3.69-3.71 (m, 3H),
3.75-3.77 (m, 3H), 3.80-3.82 (m, 2H), 3.95 (d, J=18.4 Hz, 1H), 4.18
(t, J=4.0 Hz, 2H), 4.25-4.33 (m, 3H), 4.38-4.47 (m, 4H), 4.52-4.59
(m, 2H), 4.70 (d, J=10.8 Hz, 1H), 6.26 (s, 1H), 6.84-6.91 (m, 2H),
7.00 (d, J=4.0 Hz, 1H), 7.04 (s, 1H), 7.07-7.12 (m, 1H), 7.23 (d,
J=3.2 Hz, 1H), 7.33 (d, J=7.6 Hz, 1H), 7.46-7.53 (m, 3H), 7.55-7.57
(m, 2H), 7.60-7.62 (m, 2H), 7.70 (d, J=7.6 Hz, 1H), 8.38 (t, J=6.0
Hz, 1H), 8.98 (s, 1H). Chemical Formula:
C.sub.56H.sub.54FN.sub.7O.sub.10S.sub.2; Molecular Weight:
1055.34;
[1715] The two diastereoisomerc compounds Example 256 and Example
257 could be separated by reversed phase preparative HPLC.
Synthesis of Examples 268-271 and 273
[1716] General Procedure:
##STR00718##
3-(2-chloropyrimidin-4-yl)-1-methyl-indole
##STR00719##
[1718] 2,4-dichloropyrimidine (3 g, 20.1 mmol) was dissolved in DME
(50 ml) and heated to 60.degree. C. FeCl.sub.3 (2953 mg, 22.1 mmol)
added followed by 1-methylindole (2642 mg, 20.1 mmol) and reaction
stirred at 60.degree. C. overnight. Cooled to r.t. and poured into
ice water, stirred for 30 minutes and the resulting precipitate
collected by filtration. Purified by column chromatography eluting
with DCM to provide 2.4 g (49%) of
3-(2-chloropyrimidin-4-yl)-1-methyl-indole. NMR and LC-MS conform
to structure and match literature reports.
N-(4-fluoro-2-methoxy-5-nitro-phenyl)-4-(1-methylindol-3-yl)pyrimidin-2-am-
ine
##STR00720##
[1720] 3-(2-chloropyrimidin-4-yl)-1-methyl-indole (2 g, 8.21 mmol)
and 4-fluoro-2-methoxy-5-nitro-aniline (1833 mg, 9.85 mmol) were
suspended in 2-pentanol. Para-toluenesulfonic acid (1873 mg, 9.85
mmol) was added and the reaction heated to 110.degree. C. for 2
hours under microwave conditions. The reaction mixture was allowed
to cool to r.t. and the resulting precipitate collected by
filtration, washed with 2-pentanol, triturated with acetonitrile
and dried in vacuo to a yellow solid (2453 mg, 76%). NMR and LC-MS
conform to structure and match literature reports.
2-methoxy-N4-methyl-N4-[2-(methylamino)ethyl]-N1-[4-(1-methylindol-3-yl)py-
rimidin-2-yl]-5-nitro-benzene-1,4-diamine
##STR00721##
[1722]
N-(4-fluoro-2-methoxy-5-nitro-phenyl)-4-(1-methylindol-3-yl)pyrimid-
in-2-amine (1300 mg, 3.3 mmol) dissolved in DMA (10 ml) and
N-(4-fluoro-2-methoxy-5-nitro-phenyl)-4-(1-methylindol-3-yl)pyrimidin-2-a-
mine (1.07 ml, 9.91 mmol) added followed by TEA (0.92 ml, 6.61
mmol) and the reaction heated to 140.degree. C. for 30 minutes
under microwave conditions and quenched with water (20 ml). The
reaction mixture was extracted with ethyl acetate (3.times.20 ml).
The combined organics were washed with water, dried over MgSO.sub.4
and concentrated in vacuo to a red solid. Used in subsequent steps
with no further purification.
[1723] General Procedure for Linker Introduction Via Reductive
Amination
##STR00722##
[1724]
2-methoxy-N4-methyl-N4-[2-(methylamino)ethyl]-N1-[4-(1-methylindol--
3-yl)pyrimidin-2-yl]-5-nitro-benzene-1,4-diamine (1 eq.) dissolved
in DCM and aldehyde-t-butyl ester (2.2 eq.) added followed by
sodium triacetoxyborohydride (1.2 eq.) and the reaction stirred for
1 hour at r.t. Reaction mixture concentrated in vacuo and purified
by column chromatography eluting with 0-10% methanol in DCM.
[1725] General Procedure for Nitro Group Reduction
##STR00723##
[1726] Nitro containing compound (1 eq.) suspended in 1:1
ethanol/water and iron (6 eq.) added followed by ammonium chloride
(0.75 eq.). Reaction heated to reflux for 2 hours, allowed to cool
to r.t. and filtered through celite, eluting with 10% methanol/DCM.
Filtrate extracted with DCM (3.times.), dried over MgSO4 and
concentrated in vacuo. If necessary compounds were purified by
column chromatography eluting with 0-10% methanol in DCM.
[1727] General Procedure for Aniline Acylation
##STR00724##
[1728] Acryloyl chloride (1.1 eq.) was added dropwise to a
vigorously stirred solution of aniline compound (1 eq.) and TEA (5
eq.) in DCM. Reaction stirred for 1 hour at r.t. Excess acryloyl
chloride was quenched with methanol (excess) and the reaction
mixture concentrated in vacuo. Residue purified by column
chromatography eluting with 0-10% methanol in DCM.
[1729] General Procedure for t-Butyl Ester Deprotection
##STR00725##
[1730] T-butyl ester (1 eq.) dissolved in 20% TFA in DCM and
stirred at r.t. for 1 hour. Reaction mixture concentrated in vacuo
and used immediately in the next step.
[1731] General Procedure for Coupling of E3 Recruiting Element
##STR00726##
[1732] Crude acid component (1 eq.) dissolved in DMF. HATU (1.1
eq.) added followed by TEA (5 eq.) and reaction stirred for 10
minutes. Amine coupling component (either VHL ligand,
(2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[4-(4-methylt-
hiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide or
5-(2-aminoethylamino)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione)
(1.1 eq.) added and reaction stirred overnight at r.t. Reaction
mixture diluted with water and extracted with ethyl acetate
(3.times.). Organics combined, washed with water, brine and 10%
LiCl(aq.), dried over MgSO.sub.4 and concentrated in vacuo.
Purified by chromatography eluting with 0-10% methanol in DCM.
Synthesis of Example 276
##STR00727##
[1733]
(2R,4S)-4-hydroxy-1-((S)-2-(2-(2-(2-(4-(4-((1-isopropyl-2-methyl-1H-
-imidazo[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)phenoxy)ethoxy)ethoxy)ace-
tamido)-3,3-dimethylbutanoyl)-N--((S)-1-(4-(4-methylthiazol-5-yl)phenyl)et-
hyl)pyrrolidine-2-carboxamide
[1734] Synthetic Scheme
##STR00728## ##STR00729##
1. Step--Synthesis of tert-butyl
2-(2-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethoxy)et-
hoxy)acetate
##STR00730##
[1736] The solution of tert-butyl
3-(2-(2-((methylsulfonyl)oxy)ethoxy)ethoxy)propanoate (275 mg, 0.88
mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (194
mg, 0.88 mmol) and Cs.sub.2CO.sub.3(574 mg, 1.76 mmol) was stirred
in DMF at 70.degree. C. for 1 hour, and then, after cooling to r.t,
water was added. The mixture was extracted with EA. The organic
layer was dried and concentrated. Filtered through a silica gel pad
(PE:EA=5:1) to get tert-butyl
2-(2-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethoxy)et-
hoxy)acetate (282 mg, 0.67 mmol, 76% yield).
2. Step--Synthesis of
2-(2-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethoxy)et-
hoxy)acetic acid
##STR00731##
[1738] tert-butyl
2-(2-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethoxy)et-
hoxy)acetate (282 mg, 0.67 mmol) was stirred in DCM (8 mL). TFA (2
mL) was added and continued to stir at r.t (18.degree. C.) for 1
hour. Then concentrated to get 282 mg crude product
2-(2-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethoxy)et-
hoxy)acetic acid.
3. Step--Synthesis of
(2R,4S)-1-((S)-3,3-dimethyl-2-(2-(2-(2-(4-(4,4,5,5-tetramethyl-1,3,2-diox-
aborolan-2-yl)phenoxy)ethoxy)ethoxy)acetamido)butanoyl)-4-hydroxy-N--((S)--
1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00732##
[1740] To a solution of (crude)
2-(2-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethoxy)et-
hoxy)acetic acid (157 mg, <=0.43 mmol) in DCM at r.t.
(18.degree. C.),
(2R,4S)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N--((S)-1-(4-(4-me-
thylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (150 mg,
0.35 mmol), DIPEA (135 mg, 1.04 mmol) and PyBOP (217 mg, 0.42 mmol)
were added. The mixture was stirred at r.t (18.degree. C.) for 1
hour. Water was added. The mixture was extracted with DCM. Filtered
through a silica gel pad (DCM:MeOH=20:1) to get
(2R,4S)-1-((S)-3,3-dimethyl-2-(2-(2-(2-(4-(4,4,5,5-tetramethyl-1,3,2-diox-
aborolan-2-yl)phenoxy)ethoxy)ethoxy)acetamido)butanoyl)-4-hydroxy-N--((S)--
1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
(189 mg, 0.24 mmol, 68% yield).
4. Step--Synthesis of
(2R,4S)-4-hydroxy-1-((S)-2-(2-(2-(2-(4-(4-((1-isopropyl-2-methyl-1H-imida-
zo[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)phenoxy)ethoxy)ethoxy)acetamido-
)-3,3-dimethylbutanoyl)-N--((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)py-
rrolidine-2-carboxamide
##STR00733##
[1742] A mixture of
N-(2-chloropyrimidin-4-yl)-1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin--
6-amine (28.7 mg, 0.095 mol),
(2R,4S)-1-((S)-3,3-dimethyl-2-(2-(2-(2-(4-(4,4,5,5-tetramethyl-1,3,2-diox-
aborolan-2-yl)phenoxy)ethoxy)ethoxy)acetamido)butanoyl)-4-hydroxy-N--((S)--
1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
(75.0 mg, 0.095 mol), Pd(dppf)Cl.sub.2 (13.9 mg, 0.019 mol) and CsF
(57.7 mg, 0.380 mol) in CH.sub.3CN/H.sub.2O (v/v=5/1, 3 mL) was
heated to 120.degree. C. in a microwave reactor for 30 min under
N.sub.2. After cooling to rt, the reaction was diluted with water
(5 mL) and the mixture was taken up with DCM. The organic phase was
washed with brine, dried over MgSO.sub.4, and concentrated under
vacuum. The residue was purified by chromatography (silica gel,
DCM: MeOH (10:1, v:v)) to afford the desired compound
(2R,4S)-4-hydroxy-1-((S)-2-(2-(2-(2-(4-(4-((1-isopropyl-2-methyl-1H-imida-
zo[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)phenoxy)ethoxy)ethoxy)acetamido-
)-3,3-dimethylbutanoyl)-N--((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)py-
rrolidine-2-carboxamide (7.6 mg, 9% as white solid). .sup.1H NMR
(400 MHz, CDCl3): .delta. 8.63 (s, 3H), 8.42-8.39 (m, 4H), 7.46 (d,
J=8.0 Hz, 1H), 7.35-7.31 (m, 5H), 6.99 (d, J=8.0 Hz, 2H), 6.81 (d,
J=8.0 Hz, 1H), 5.09-5.05 (m, 1H), 4.71-4.67 (m, 2H), 4.49-4.48 (m,
1H), 4.31-4.30 (m, 1H), 4.18 (s, 2H), 4.08-4.07 (m, 1H), 4.01 (s,
1H), 3.96 (s, 1H), 3.81-3.79 (m, 2H), 3.75-3.73 (m, 4H), 3.62 (dd,
J=5.6 Hz, 1H), 2.65 (s, 3H), 2.42-2.40 (m, 4H), 2.09-2.06 (t, J=8.0
Hz, 1H), 1.73 (d, J=8.0 Hz, 6H), 1.38 (d, J=8.0 Hz, 3H), 1.08 (s,
9H). LC-MS: (ES.sup.+): m/z 933.4 [M+H]. t.sub.R=3.389 min Chemical
Formula: C49H60N10O7S; Molecular Weight: 933.13
Synthesis of Example 282
(2S,4R)-4-hydroxy-1-((S)-2-(2-(3-(3-(9-(4-((1-isopropyl-2-methyl-1H-imidaz-
o[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)-3,9-diazaspiro[5.5]undecan-3-yl-
)propoxy)propoxy)acetamido)-3,3-dimethylbutanoyl)-N--((S)-1-(4-(4-methylth-
iazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00734##
[1744] Synthetic Scheme
##STR00735##
Experimental Section
1. Step--Synthesis of Tert-butyl
9-(4-((1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)pyrimidin-
-2-yl)-3,9-diazaspiro[5.5]undecane-3-carboxylate
##STR00736##
[1746] A mixture of
N-(2-chloropyrimidin-4-yl)-1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin--
6-amine (150 mg, 0.47 mmol), tert-butyl
3,9-diazaspiro[5.5]undecane-3-carboxylate (125 mg, 0.49 mmol) and
Et.sub.3N (142 mg, 1.4 mmol) in PrOH (5 mL) was stirred at
150.degree. C. for 1 h. After cooling to rt, the solvent was
removed under vacuum. The residue was purified by silica gel column
chromatography with MeOH/DCM (1:20) as eluent to afford the desired
product Tert-butyl
9-(4-((1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)pyrimidin-
-2-yl)-3,9-diazaspiro[5.5]undecane-3-carboxylate as a yellow solid
(200 mg, 0.38 mmol, 80.8% yield). Chemical Formula:
C.sub.28H.sub.40N.sub.8O.sub.2; Molecular Weight: 520.68. LC-MS:
(ES.sup.+): m/z 521.3 [M+H]. t.sub.R=3.33 min
2. Step--Synthesis of
N-(2-(3,9-Diazaspiro[5.5]undecan-3-yl)pyrimidin-4-yl)-1-isopropyl-2-methy-
l-1H-imidazo[4,5-c]pyridin-6-amine hydrochloride
##STR00737##
[1748] To a solution of Tert-butyl
9-(4-((1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)pyrimidin-
-2-yl)-3,9-diazaspiro[5.5]undecane-3-carboxylate (200 mg, 0.38
mmol) in MeOH (8 mL) was added 6N HCl in dioxane (4 ml, 24 mmol) at
rt. After stirring 30 min, the mixture was concentrated under
vacuum to afford the crude
N-(2-(3,9-Diazaspiro[5.5]undecan-3-yl)pyrimidin-4-yl)-1-isopropyl-2-
-methyl-1H-imidazo[4,5-c]pyridin-6-amine hydrochloride as a yellow
solid (200 mg), which was used in next step without further
purification. Chemical Formula: C.sub.23H.sub.33ClN.sub.8;
Molecular Weight: 457.02 LC-MS: (ES.sup.+): m/z 421.3 [M+H].
t.sub.R=2.19 min
3. Step--Synthesis of Tert-butyl
2-(3-(3-(9-(4-((1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)-
pyrimidin-2-yl)-3,9-diazaspiro[5.5]undecan-3-yl)propoxy)propoxy)acetate
##STR00738##
[1750] To a mixture of
N-(2-(3,9-Diazaspiro[5.5]undecan-3-yl)pyrimidin-4-yl)-1-isopropyl-2-methy-
l-1H-imidazo[4,5-c]pyridin-6-amine hydrochloride (190 mg crude,
<=0.38 mmol), tert-butyl
2-(3-(3-((methylsulfonyl)oxy)propoxy)propoxy)acetate (186 mg, 0.57
mmol) in DMF (10 ml) was added K.sub.2CO.sub.3 (209 mg, 1.51 mmol).
The resulting solution was stirred at 80.degree. C. for 1 h. After
cooling to rt, the reaction was diluted with 10 mL water, and the
mixture was extracted with ethyl acetate (2.times.30 mL). The
combined organic layers were washed with brine (10 mL), dried over
Na.sub.2SO.sub.4 and filtered. The solvent was evaporated under
reduced pressure. The residue was purified by pre-TLC
(DCM:MeOH=20:1) to afford the desired compound Tert-butyl
2-(3-(3-(9-(4-((1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)-
pyrimidin-2-yl)-3,9-diazaspiro[5.5]undecan-3-yl)propoxy)propoxy)acetate
as a white solid (65 mg, 0.10 mmol, 26.3% yield). LC-MS:
(ES.sup.+): m/z 651.4 [M+H]. t.sub.R=2.79 min Chemical Formula:
C.sub.35H.sub.54N.sub.8O.sub.4; Molecular Weight: 650.87
4. Step--Synthesis of
2-(3-(3-(9-(4-((1-Isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)-
pyrimidin-2-yl)-3,9-diazaspiro[5.5]undecan-3-yl)propoxy)propoxy)acetic
acid
##STR00739##
[1752] To a solution of Tert-butyl
2-(3-(3-(9-(4-((1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)-
pyrimidin-2-yl)-3,9-diazaspiro[5.5]undecan-3-yl)propoxy)propoxy)acetate
(65 mg, 0.10 mmol) in DCM (2 mL) were added TFA (2 ml), After
stirring 1 h, the solvent was removed in vacuo to afford the crude
desired product
2-(3-(3-(9-(4-((1-Isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)-
pyrimidin-2-yl)-3,9-diazaspiro[5.5]undecan-3-yl)propoxy)propoxy)acetic
acid (100 mg, crude), which was used into next reaction without
further purification. LC-MS: (ES.sup.+): m/z 595.4 [M+H].
t.sub.R=2.36 min Chemical Formula: C.sub.31H.sub.46N.sub.8O.sub.4;
Molecular Weight: 594.76
5. Step--Synthesis of
(2S,4R)-4-Hydroxy-1-((S)-2-(2-(3-(3-(9-(4-((1-Isopropyl-2-methyl-1H-imida-
zo[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)-3,9-diazaspiro[5.5]undecan-3-y-
l)propoxy)propoxy)acetamido)-3,3-dimethylbutanoyl)-N--((S)-1-(4-(4-methylt-
hiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00740##
[1754] To a solution of
2-(3-(3-(9-(4-((1-Isopropyl-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)amino)-
pyrimidin-2-yl)-3,9-diazaspiro[5.5]undecan-3-yl)propoxy)propoxy)acetic
acid (100 mg, crude, <=0.10 mmol),
(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N--((S)-1-(4-(4-me-
thylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (72 mg,
0.15 mmol), DIPEA (77 mg, 0.60 mmol) in DCM (5 mL) was added PyBop
(78 mg, 0.15 mmol). The resulting solution was stirred at rt for 1
h. After quenched with water (10 mL), the mixture was extracted
with DCM (2.times.30 mL). The combined organic layers were washed
with brine (10 mL), dried over Na.sub.2SO.sub.4, filtered and
concentrated under vacuum, The residue was purified by prep-HPLC to
afford
(2S,4R)-4-Hydroxy-1-((S)-2-(2-(3-(3-(9-(4-((1-Isopropyl-2-methyl-1H-imida-
zo[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)-3,9-diazaspiro[5.5]undecan-3-y-
l)propoxy)propoxy)acetamido)-3,3-dimethylbutanoyl)-N--((S)-1-(4-(4-methylt-
hiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide as a white solid
(45 mg, 0.044 mmol, 44.0% yield, 2 steps). .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 13.15 (bs, 1H), 11.30 (s, 1H), 8.88 (s, 1H),
8.65 (s, 1H), 8.54 (s, 1H), 7.99 (s, 1H), 7.44 (d, J=7.2 Hz, 1H),
7.42 (s, 4H), 8.67 (d, J=8.7 Hz, 1H), 6.69 (s, 1H), 5.08-5.10 (m,
1H), 4.71-4.77 (m, 2H), 4.60-4.62 (m, 1H), 4.51 (s, 1H), 3.91-4.02
(m, 6H), 3.53-3.64 (m, 9H), 3.10-3.13 (m, 2H), 2.85-2.88 (m, 2H),
2.88 (s, 3H), 2.55 (m, 3H), 2.33-2.34 (m, 1H), 2.10-2.12 (m, 1H),
1.86-2.00 (m, 11H), 1.69 (d, J=6.8 Hz, 6H), 1.59-1.61 (m, 2H), 1.48
(d, J=6.8 Hz, 3H), 1.06 (s, 9H). LC-MS: (ES.sup.+): m/z 1021.5
[M+H]. t.sub.R=2.84 min Chemical Formula:
C.sub.54H.sub.76N.sub.12O.sub.6S; Molecular Weight: 1021.34
Synthesis of Example 283
2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)ox-
y)ethoxy)ethoxy)ethoxy)propyl)-1-oxoisoindolin-2-yl)-2-(5-fluoro-2-hydroxy-
phenyl)-N-(thiazol-2-yl)acetamide
##STR00741##
[1756] Synthetic Scheme:
##STR00742##
1. Step--Synthesis of
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-
-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)propyl)-1-oxoisoi-
ndolin-2-yl)-N-(thiazol-2-yl)acetamide
##STR00743##
[1758] A mixture of palladium on carbon (10%, 40 mg) and
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-
-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)prop-1-yn-1-yl)-l-
-oxoisoindolin-2-yl)-N-(thiazol-2-yl)acetamide (80 mg, 0.087 mmol)
in methanol (20 ml) was stirred at room temperature overnight under
hydrogen atmosphere (hydrogen balloon). TLC showed the reaction was
complete. Palladium on carbon was removed through filtration and
washed with methanol (5 ml.times.2). The combined filtrates were
concentrated under reduced pressure to afford
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-
-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)propyl)-1-oxoisoi-
ndolin-2-yl)-N-(thiazol-2-yl)acetamide (70 mg, crude) as yellow
oil. LC_MS: (ES.sup.+): m/z 920.5 [M+H].sup.+. t.sub.R=3.010
min.
2. Step--Synthesis of
2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)o-
xy)ethoxy)ethoxy)ethoxy)propyl)-1-oxoisoindolin-2-yl)-2-(5-fluoro-2-hydrox-
yphenyl)-N-(thiazol-2-yl)acetamide
##STR00744##
[1760] To a stirred solution of
2-(2-(benzyloxy)-5-fluorophenyl)-2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-
-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)propyl)-1-oxoisoi-
ndolin-2-yl)-N-(thiazol-2-yl)acetamide (70 mg, 0.076 mmol) in
dichloromethane (8 ml) was drop wise boron tribromide (95 mg, 0.38
mmol) in dichloromethane (1 ml) at -70.degree. C. dropwise. The
resulting mixture was stirred at -70.degree. C. for 1 hour. TLC
showed the reaction was complete. The reaction mixture was diluted
with dichloromethane (15 ml) and quenched with saturated sodium
bicarbonate solution (8 ml) at -40.degree. C. The organic layer was
collected and the aqueous layer was extracted with dichloromethane
(15 ml.times.2). The combined organic layers were washed with brine
(20 ml), dried over anhydrous sodium sulfate, and concentrated
under reduced pressure to give a crude residue which was purified
by preparative TLC (eluted with 5% methanol in dichloromethane) to
afford
2-(7-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)o-
xy)ethoxy)ethoxy)ethoxy)propyl)-1-oxoisoindolin-2-yl)-2-(5-fluoro-2-hydrox-
yphenyl)-N-(thiazol-2-yl)acetamide (25 mg, yield 27%) as white
solid. LC_MS: (ES.sup.+): m/z 830.4 [M+H].sup.+. t.sub.R=2.645 min.
.sup.1H NMR (400 MHz, DMSO-d6): .delta. 1.97-2.03 (m, 4H),
2.54-2.68 (m, 1H), 3.07 (t, J=7.2H, 1H), 3.38-3.41 (m, 4H),
3.47-3.48 (m, 2H), 3.52-3.56 (m, 4H), 3.58-3.60 (m, 2H), 3.78 (t,
J=4.0 Hz, 2H), 3.89-3.93 (m, 1H), 4.29 (t, J=4.0 Hz, 2H), 4.52-4.56
(m, 1H), 5.09-5.14 (m, 1H), 5.36 (d, J=4.8 Hz, 1H), 6.26 (s, 1H),
6.82-6.85 (m, 1H), 6.88-6.91 (m, 1H), 7.07-7.12 (m, 1H), 7.24-7.26
(m, 2H), 7.34-7.37 (m, 2H), 7.43-7.48 (m, 3H), 7.80 (d, J=12.0 Hz,
1H), 8.32 (s, 1H), 11.14 (s, 1H). Chemical Formula:
C.sub.41H.sub.40FN.sub.5O.sub.11S; Molecular Weight: 829.85;
Synthesis of Example 286
2-(2,6-dioxopiperidin-3-yl)-5-(2-(4-(4-(1-isopropyl-6-((2-(4-methoxypiperi-
din-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)benzyl)pipera-
zin-1-yl)ethoxy)isoindoline-1,3-dione
##STR00745##
[1762] Synthetic Route:
##STR00746##
Experimental Section
1. Step--Synthesis of
4-((4-(2-((Tert-butyldimethylsilyl)oxy)ethyl)piperazin-1-yl)methyl)benzoi-
c acid
##STR00747##
[1764] To a solution of
1-(2-((tert-butyldimethylsilyl)oxy)ethyl)piperazine (500 mg, 2.00
mmol) in MeOH (10 mL) were added 4-formylbenzoic acid (300 mg, 2.00
mmol) and NaBH.sub.3CN (151 mg, 2.4 mmol). The solution was stirred
at rt for 3 h. Then the solution was diluted with water (10 mL).
The resulting mixture was extracted with DCM (2.times.30 mL). The
combined organic layers were washed with brine (20 mL). The organic
phase was dried over anhydrous sodium sulfate, filtered and
concentrated under vacuum. The residue was purified by silica gel
column chromatography (DCM:MeOH=10:1) to afford
4-((4-(2-((Tert-butyldimethylsilyl)oxy)ethyl)piperazin-1-yl)methyl)benzoi-
c acid as a white solid (120 mg, 0.31 mmol, 15.8% yield). Chemical
Formula: C.sub.20H.sub.34N.sub.2O.sub.3Si; Molecular Weight:
378.59.
2. Step--Synthesis of
4-((4-(2-((Tert-butyldimethylsilyl)oxy)ethyl)piperazin-1-yl)methyl)-N-(4--
(isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)pyrid-
in-3-yl)benzamide
##STR00748##
[1766] To a solution of
N4-isopropyl-N2-(2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)pyridine-2,4,5-
-triamine (400 mg, 1.12 mmol) and
4-((4-(2-((Tert-butyldimethylsilyl)oxy)ethyl)piperazin-1-yl)methyl)benzoi-
c acid (677 mg, 1.80 mmol) in DCM (20 mL) were added DIPEA (578 mg,
4.48 mmol) and HATU (851 mg, 2.24 mmol) at rt. The solution was
stirred at rt overnight. The solution was diluted with water (5 mL)
and the resulting mixture was extracted with DCM (2.times.20 mL).
The combined organic layers were washed with brine (10 mL). The
organic phase was dried over anhydrous sodium sulfate, filtered and
concentrated under vacuum. The residue was purified by silica gel
column chromatography (DCM:MeOH=10:1) to afford
4-((4-(2-((Tert-butyldimethylsilyl)oxy)ethyl)piperazin-1-yl)met-
hyl)-N-(4-(isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)a-
mino)pyridin-3-yl)benzamide as a white solid (100 mg, 0.14 mmol,
12.4% yield). Chemical Formula: C.sub.38H.sub.59N.sub.9O.sub.3Si;
Molecular Weight: 718.03
3. Step--Synthesis of
2-(4-(4-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-
-1H-imidazo[4,5-c]pyridin-2-yl)benzyl)piperazin-1-yl)ethylacetate
##STR00749##
[1768] A solution of
4-((4-(2-((Tert-butyldimethylsilyl)oxy)ethyl)piperazin-1-yl)methyl)-N-(4--
(isopropylamino)-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)pyrid-
in-3-yl)benzamide (150 mg, 0.21 mmol) in CH.sub.3COOH (5 ml) was
stirred at 150.degree. C. for 10 h under microwave. Then the
reaction mixture was evaporated under reduced pressure. The pH of
the solution was adjusted to 8 with saturated NaHCO.sub.3. The
resulting mixture was extracted with DCM (2.times.10 mL). The
combined organic layers were washed with brine (5 mL). The organic
phase was dried over anhydrous sodium sulfate, filtered and
concentrated under vacuum. The residue was purified by prep-TLC
(DCM:MeOH=10:1) to afford
2-(4-(4-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-
-1H-imidazo[4,5-c]pyridin-2-yl)benzyl)piperazin-1-yl)ethyl acetate
as a white solid (42 mg, 0.064 mmol, 31.8% yield). Chemical
Formula: C.sub.34H.sub.45N.sub.9O.sub.3; Molecular Weight:
627.79
4. Step--Synthesis of
2-(4-(4-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-
-1H-imidazo[4,5-c]pyridin-2-yl)benzyl)piperazin-1-yl)ethan-1-ol
##STR00750##
[1770] To a solution of
2-(4-(4-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-
-1H-imidazo[4,5-c]pyridin-2-yl)benzyl)piperazin-1-yl)ethyl acetate
(40 mg, 0.064 mmol) in THF (2 mL) and H.sub.2O (1 ml) was added
LiOH--H.sub.2O (16 mg, 0.38 mmol). The mixture was stirred at rt
for 1 h. The resulting reaction mixture was extracted with EA
(2.times.5 mL). The combined organic layers were washed with brine
(5 mL). The organic phase was dried over anhydrous sodium sulfate,
filtered and concentrated under vacuum to afford
2-(4-(4-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl-
)amino)-1H-imidazo[4,5-c]pyridin-2-yl)benzyl)piperazin-1-yl)ethan-1-ol
(40 mg, crude), which was used in the next step without further
purification. LC-MS: (ES.sup.+): m/z 586.3 [M+H].sup.+.
t.sub.R=2.51 min Chemical Formula: C.sub.32H.sub.43N.sub.9O.sub.2;
Molecular Weight: 585.76
5.
Step--2-(2,6-Dioxopiperidin-3-yl)-5-(2-(4-(4-(1-isopropyl-6-((2-(4-meth-
oxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)benz-
yl)piperazin-1-yl)ethoxy)isoindoline-1,3-dione
##STR00751##
[1772] To a mixture of
2-(4-(4-(1-Isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-
-1H-imidazo[4,5-c]pyridin-2-yl)benzyl)piperazin-1-yl)ethan-1-ol (30
mg, 0.051 mmol),
2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (18 mg,
0.066 mmol) and PPh.sub.3 (78 mg, 0.31 mmol) in THF (5 mL) was
added DIAD (60 mg, 0.31 mmol) dropwise at 45.degree. C. The
solution was stirred at 45.degree. C. for 10 min. Then the reaction
mixture was diluted with water (10 mL). The mixture was extracted
with DCM (2.times.10 mL). The combined organic layers were washed
with brine (5 mL). The organic phase was dried over anhydrous
sodium sulfate, filtered and concentrated under vacuum. The residue
was purified by prep-TLC to afford
2-(2,6-Dioxopiperidin-3-yl)-5-(2-(4-(4-(1-isopropyl-6-((2-(4-metho-
xypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)benzy-
l)piperazin-1-yl)ethoxy)isoindoline-1,3-dione as a white solid (4.5
mg, 0.0053 mmol, 10.0% yield). .sup.1H NMR (400 MHz, CD.sub.3OD):
.delta. 8.65 (s, 1H), 8.57 (s, 1H), 7.95 (d, J=6.0 Hz, 1H), 7.80
(d, J=8.0 Hz, 1H), 7.65 (d, J=8.0 Hz, 2H), 7.59 (d, J=8.0 Hz, 2H),
7.58 (s, 1H), 7.43 (d, J=2.0 Hz, 1H), 6.32 (d, J=5.6 Hz, 1H),
5.31-5.34 (m, 2H), 5.05-5.15 (m, 1H), 4.29-4.32 (m, 4H), 3.67 (s,
2H), 3.34-3.36 (m, 4H), 3.30 (m, 3H), 2.82-2.83 (m, 3H), 2.65-2.80
(m, 4H), 2.15-2.20 (m, 2H), 2.10-2.12 (m, 1H), 1.95-1.98 (m, 6H),
1.68 (d, J=6.8 Hz, 6H). LC-MS: (ES.sup.+): m/z 842.4 [M+H].sup.+.
t.sub.R=2.71 min Chemical Formula: C.sub.45H.sub.51N.sub.11O.sub.6;
Molecular Weight: 841.97
Synthesis of Example 291
(2S,4R)-1-((S)-2-(Tert-butyl)-14-(4-((1-(4-((1-isopropyl-2-methyl-1H-imida-
zo[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)piperidin-4-yl)oxy)phenyl)-4-ox-
o-6,9,12-trioxa-3-azatetradecanoyl)-4-hydroxy-N--((S)-1-(4-(4-methylthiazo-
l-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00752##
[1774] The synthesis of example 291 followed the route and methods
described for example 282 using the building blocks shown in the
scheme below.
##STR00753##
[1775] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 8.66 (s, 1H),
8.61 (s, 1H), 8.30 (s, 1H), 8.03 (d, J=5.6 Hz, 1H), 7.32-7.41 (m,
7H), 7.14 (d, J=8.4 Hz, 2H), 6.87 (d, J=8.4 Hz, 2H), 6.09 (d, J=5.6
Hz, 1H), 5.10 (t, J=7.2 Hz, 1H), 4.73-4.76 (m, 1H), 4.62-4.66 (m,
1H), 4.50-4.52 (m, 3H), 4.22-4.28 (m, 2H), 4.12-4.15 (m, 1H),
3.95-4.10 (m, 2H), 3.63-3.75 (m, 13H), 2.84 (t, J=7.2 Hz, 2H), 2.61
(s, 3H), 2.60-2.62 (m, 1H), 2.52 (m, 3H), 1.98-2.04 (m, 4H),
1.80-1.85 (m, 2H), 1.63 (d, J=6.8 Hz, 6H), 1.53 (d, J=6.4 Hz, 3H),
1.07 (s, 9H).
[1776] LC-MS: (ES.sup.+): m/z 1060.5 [M+H]. t.sub.R=3.42 min
Chemical Formula: C.sub.56H.sub.73N.sub.11O.sub.8S; Molecular
Weight: 1060.33.
Synthesis of Example 292
(2S,4R)-4-hydroxy-1-((S)-2-(2-(3-(3-(4-(2-(1-(4-((1-isopropyl-2-methyl-1H--
imidazo[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)-4-methoxypiperidin-4-yl)e-
thyl)piperazin-1-yl)propoxy)propoxy)acetamido)-3,3-dimethylbutanoyl)-N--((-
S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00754##
[1778] The synthesis of example 292 followed the general strategy
and methods used for examples 78 and 282, using the building blocks
shown in the scheme below.
##STR00755##
(2S,4R)-4-Hydroxy-1-((S)-2-(2-(3-(3-(4-(2-(1-(4-((1-isopropyl-2-methyl-H--
imidazo[4,5-c]pyridin-6-yl)amino)pyrimidin-2-yl)-4-methoxypiperidin-4-yl)e-
thyl)piperazin-1-yl)propoxy)propoxy)acetamido)-3,3-dimethylbutanoyl)-N--((-
S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide
##STR00756##
[1780] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 8.59 (s, 1H),
8.53 (s, 1H), 8.31 (s, 1H), 7.94 (d, J=5.6 Hz, 1H), 7.69 (s, 1H),
7.35-7.36 (m, 1H), 7.29-7.33 (m, 4H), 7.15-7.17 (m, 1H), 5.98 (d,
J=5.2 Hz, 1H), 4.95-5.05 (m, 1H), 4.36-4.38 (m, 2H), 4.32-4.35 (m,
4H), 3.95-4.02 (m, 1H), 3.86 (d, J=4.0 Hz, 2H), 3.51-3.54 (m, 3H),
3.42-3.43 (m, 4H), 3.25-3.27 (m, 2H), 3.16 (s, 3H), 2.54 (s, 3H),
2.43-2.46 (m, 9H), 2.35-2.38 (m, 6H), 1.98-2.02 (m, 2H), 1.76-1.78
(m, 5H), 1.67-1.69 (m, 3H), 1.65 (d, J=6.8 Hz, 6H), 1.48-1.51 (m,
3H), 1.40 (d, J=6.8 Hz, 3H), 0.98 (s, 9H). LC-MS: (ES.sup.+): m/z
1094.3 [M+H].sup.+. t.sub.R=2.98 min Chemical Formula:
C.sub.57H.sub.83N.sub.13O.sub.7S; Molecular Weight: 1094.43
Synthesis of Example 301
2-(2,6-dioxopiperidin-3-yl)-5-(4-(3-(4-(4-(2-((1-isopropyl-6-((2-(4-methox-
ypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)methox-
y)ethyl)piperazin-1-yl)phenoxy)propyl)piperazin-1-yl)isoindoline-1,3-dione
##STR00757##
[1782] Synthetic Route:
##STR00758## ##STR00759##
1. Step--Synthesis of
2-chloro-N-isopropyl-5-nitro-pyridin-4-amine
##STR00760##
[1784] To a solution of 2,4-dichloro-5-nitro-pyridine (14 g, 72.5
mmol, 1 eq) and propan-2-amine (4.29 g, 72.5 mmol, 1 eq) in
tetrahydrofuran (100 mL) was added triethylamine (14.68 g, 145.09
mmol, 2 eq), the mixture was stirred at 25.degree. C. for 6 hours.
Thin Layer Chromatography (petroleum ether/ethyl acetate=5/1)
showed the starting material was consumed completely and one major
new spot with higher polarity was detected. The mixture was diluted
with water (200 mL), then extracted with ethyl acetate (100
mL.times.3), the organic layer was dried over anhydrous sodium
sulfate, filtered and concentrated under reduced pressure to afford
2-chloro-N-isopropyl-5-nitro-pyridin-4-amine (15.5 g, 71.88 mmol,
99% yield) as a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.: 9.02 (s, 1H), 8.08 (s, 1H), 6.74 (s, 1H), 3.79-3.87 (m,
1H), 1.37 (d, J=6.4 Hz, 6H). Chemical Formula:
C.sub.8H.sub.10ClN.sub.3O.sub.2, Molecular Weight: 215.64
2. Step--Synthesis of
6-chloro-N-4-isopropyl-pyridine-3,4-diamine
##STR00761##
[1786] To a mixture of 2-chloro-N-isopropyl-5-nitro-pyridin-4-amine
(10 g, 46.37 mmol, 1 eq) and iron (6.47 g, 115.94 mmol, 2.5 eq) in
methanol (80 mL) was added ammonium chloride (6.20 g, 115.94 mmol,
2.5 eq) in water (20 mL). The mixture was stirred at 65.degree. C.
for 12 hours. Thin Layer Chromatography (petroleum ether/ethyl
acetate=1/1) showed the starting material was consumed completely
and one major new spot was formed. The mixture was concentrated
under reduced pressure to give a residue. The residue was diluted
with ethyl acetate (30 mL), filtered and the filtrate was
concentrated under reduced pressure to give a residue. The residue
was purified by silica gel chromatography (petroleum ether/ethyl
acetate=10/1 to 1/1) to afford
6-chloro-N-4-isopropyl-pyridine-3,4-diamine (5.6 g, 30.16 mmol, 65%
yield) as a gray solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.:
7.62 (s, 1H), 6.43 (s, 1H), 4.23 (s, 1H), 3.56-3.67 (m, 1H), 3.02
(s, 2H), 1.25 (d, J=6.4 Hz, 6H). Chemical Formula:
C.sub.8H.sub.12ClN.sub.3, Molecular Weight: 185.65
3. Step--Synthesis of 6-chloro-2-(chloromethyl)-1-isopropyl-imidazo
[4,5-c]pyridine
##STR00762##
[1788] To a solution of 6-chloro-N4-isopropyl-pyridine-3,4-diamine
(1.3 g, 7.00 mmol, 1 eq) in 2-chloro-1,1,1-trimethoxy-ethane (4.60
g, 29.76 mmol, 4.0 mL, 4.25 eq) was added p-toluenesulfonic acid
monohydrate (133 mg, 0.70 mmol, 0.1 eq), the mixture was stirred at
100.degree. C. for 1 hour. Thin layer chromatography (petroleum
ether/ethyl acetate=3/1) showed one major new spot with lower
polarity was detected. The mixture was concentrated under reduced
pressure. The residue was purified by silica gel chromatography
(petroleum ether/ethyl acetate=3/1) to afford
6-chloro-2-(chloromethyl)-1-isopropyl-imidazo[4,5-c] pyridine (0.82
g, 3.36 mmol, 48% yield) as a light yellow solid. .sup.1H NMR: (400
MHz, CDCl.sub.3) .delta.: 8.76 (s, 1H), 7.98 (s, 1H), 5.14 (s, 2H),
4.85-4.93 (m, 1H), 1.58 (d, J=6.8 Hz, 6H). Chemical Formula:
C.sub.10H.sub.11Cl.sub.2N.sub.3, Molecular Weight: 244.12
4. Step--Synthesis of
6-chloro-2-(2,2-dimethoxyethoxymethyl)-1-isopropyl-imidazo[4,5-c]pyridine
##STR00763##
[1790] To a solution of 2,2-dimethoxyethanol (469 mg, 4.42 mmol,
1.5 eq) in N,N-dimethylformamide (20 mL) was added sodium hydride
(177 mg, 4.42 mmol, 60% purity, 1.5 eq), the mixture was stirred at
50.degree. C. for 1 hour, then
6-chloro-2-(chloromethyl)-1-isopropyl-imidazo[4,5-c]pyridine (0.72
g, 2.95 mmol, 1 eq) was added, the mixture was stirred at
50.degree. C. for 2 hours. The desired MS was detected by LCMS. The
mixture was diluted with water (100 mL), then extracted with ethyl
acetate (50 mL.times.3), the organic layer was washed with brine
(100 mL.times.2), dried over anhydrous sodium sulfate, filtered and
concentrated under reduced pressure. The residue was purified by
silica gel chromatography (petroleum ether/ethyl acetate=10/1 to
1/1) to afford
6-chloro-2-(2,2-dimethoxyethoxymethyl)-1-isopropyl-imidazo[4,5-c]pyridine
(0.7 g, 2.23 mmol, 75% yield) as a light yellow oil. LCMS: MS (ESI)
m/z: 314.1 [M+1]+. .sup.1H NMR: (400 MHz, CDCl.sub.3) .delta.: 8.80
(d, J=0.8 Hz, 1H), 7.49 (d, J=0.8 Hz, 1H), 4.90-4.97 (m, 1H), 4.85
(s, 2H), 4.51 (t, J=5.2 Hz, 1H), 3.57 (d, J=5.2 Hz, 2H), 3.37 (s,
6H), 1.64 (d, J=6.8 Hz, 6H). Chemical Formula:
C.sub.14H.sub.20ClN.sub.3O.sub.3, Molecular Weight: 313.78
5. Step--Synthesis of
2-(2,2-dimethoxyethoxymethyl)-1-isopropyl-N-[2-(4-methoxy-1-piperidyl)pyr-
imidin-4-yl]imidazo[4,5-c]pyridin-6-amine
##STR00764##
[1792] To a solution of
6-chloro-2-(2,2-dimethoxyethoxymethyl)-1-isopropyl-imidazo[4,5-c]pyridine
(500 mg, 1.59 mmol, 1 eq) and
2-(4-methoxy-1-piperidyl)pyrimidin-4-amine (331 mg, 1.59 mmol, 1
eq) in 1,4-dioxane (10 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (152 mg,
0.32 mmol, 0.2 eq), tris(dibenzylideneacetone)dipalladium (146 mg,
0.16 mmol, 0.1 eq) and cesium carbonate (1.04 g, 3.19 mmol, 2 eq),
the mixture was degassed and purged with nitrogen several times,
then stirred at 100.degree. C. for 3 hours under nitrogen
atmosphere. The desired MS was detected by LCMS. The mixture was
concentrated under reduced pressure. The residue was purified by
silica gel chromatography (dichloromethane/methanol=20/1) to afford
2-(2,2-dimethoxyethoxymethyl)-1-isopropyl-N-[2-(4-methoxy-1-piperidyl)pyr-
imidin-4-yl]imidazo[4,5-c]pyridin-6-amine (0.7 g, 1.44 mmol, 90%
yield) as a light yellow oil. LCMS: MS (ESI) m/z: 486.2
[M+1].sup.+. .sup.1H NMR: (400 MHz, CDCl.sub.3) .delta.: 8.70 (s,
1H), 8.46 (s, 1H), 8.05 (d, J=5.6 Hz, 1H), 7.37 (s, 1H), 6.04 (d,
J=5.6 Hz, 1H), 4.93-4.98 (m, 1H), 4.84 (s, 2H), 4.51 (t, J=5.2 Hz,
1H), 4.36-4.41 (m, 1H), 3.57 (d, J=5.2 Hz, 2H), 3.44-3.54 (m, 4H),
3.42 (s, 3H), 3.38 (s, 6H), 1.95-2.03 (m, 2H), 1.68 (d, J=7.2 Hz,
6H), 1.64-1.66 (m, 2H). Chemical Formula:
C.sub.24H.sub.35N.sub.7O.sub.4, Molecular Weight: 485.58
6. Step--Synthesis of 2-[[1-isopropyl-6-[[2-(4-methoxy-1-piperidyl)
pyrimidin-4-yl]amino]imidazo[4,5-c]pyridin-2-yl]methoxy]acetaldehyde
##STR00765##
[1794] To a solution of
2-(2,2-dimethoxyethoxymethyl)-1-isopropyl-N-[2-(4-methoxy-1-piperidyl)
pyrimidin-4-yl]imidazo[4,5-c]pyridin-6-amine (0.6 g, 1.24 mmol, 1
eq) in tetrahydrofuran (20 mL) was added sulfuric acid (2 molar,
aqueous, 20 mL, 40 mmol, 32 equiv.), the mixture was stirred at
100.degree. C. for 30 minutes. Thin layer chromatography
(dichloromethane/methanol=20/1) showed the starting material was
consumed completely and one major new spot with larger polarity was
detected. The pH of the mixture was adjusted to 8 by saturated
solution of sodium carbonate, then extracted with ethyl acetate (30
mL.times.3), the organic layer was dried over anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The
residue purified by silica gel chromatography
(dichloromethane/methanol=50/1 to 20/1) to afford
2-[[1-isopropyl-6-[[2-(4-methoxy-1-piperidyl)pyrimidin-4-yl]amino]imidazo-
[4,5-c]pyridin-2-yl]methoxy]acetaldehyde (0.5 g, 1.14 mmol, 92%
yield) as a light yellow oil. .sup.1H NMR: (400 MHz, CDCl.sub.3)
.delta.: 9.68 (s, 1H), 8.70 (s, 1H), 8.49 (s, 1H), 8.06 (d, J=5.6
Hz, 1H), 7.50-7.53 (m, 1H), 6.05 (d, J=5.6 Hz, 1H), 4.98-5.05 (m,
1H), 4.90 (s, 2H), 4.38-4.40 (m, 2H), 4.27 (s, 2H), 3.51-3.54 (m,
2H), 3.47-3.49 (m, 1H), 3.42 (s, 3H), 1.96-2.00 (m, 2H), 1.70 (d,
J=7.2 Hz, 6H), 1.65-1.67 (m, 2H). Chemical Formula:
C.sub.22H.sub.29N.sub.7O.sub.3, Molecular Weight: 439.51
7. Step--Synthesis of tert-butyl
4-[4-(3-bromopropoxy)phenyl]piperazine-1-carboxylate
##STR00766##
[1796] To a solution of tert-butyl
4-(4-hydroxyphenyl)piperazine-1-carboxylate (500 mg, 1.80 mmol, 1
eq) in acetone (10 mL) was added potassium carbonate (745 mg, 5.39
mmol, 3 eq) and 1,3-dibromopropane (1.09 g, 5.39 mmol, 3 eq). The
mixture was stirred at 80.degree. C. for 12 hr. LCMS showed the
reaction was completed. The reaction mixture was diluted with water
(30 mL) and extracted with ethyl acetate (10 mL.times.2). The
combined organic phase was washed with saturated brine (10
mL.times.2), dried with anhydrous sodium sulfate, filtered and
concentrated in vacuum. The residue was purified by silica gel
chromatography (Petroleum ether/Ethyl acetate=30:1 to 10:1).
tert-butyl 4-[4-(3-bromopropoxy)phenyl]piperazine-1-carboxylate
(620 mg, 1.55 mmol, 86% yield) was obtained as a white solid. LCMS:
MS (ESI) m/z: 399.0 [M+1].sup.+. Chemical Formula:
C.sub.17H.sub.27N.sub.2O.sub.3Br, Molecular Weight: 398.12
8. Step--Synthesis of tert-butyl
4-[4-[3-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-
-1-yl]propoxy]phenyl]piperazine-1-carboxylate
##STR00767##
[1798] To a solution of tert-butyl
4-[4-(3-bromopropoxy)phenyl]piperazine-1-carboxylate (400 mg, 1.00
mmol, 1 eq) and
2-(2,6-dioxo-3-piperidyl)-5-piperazin-1-yl-isoindoline-1,3-dion- e
(379 mg, 1.00 mmol, 1 eq, HCl) in acetonitrile (10 mL) was added
potassium iodide (17 mg, 0.10 mmol, 0.1 eq) and
diisopropylethylamine (518 mg, 4.01 mmol, 0.7 mL, 4 eq). The
mixture was stirred at 100.degree. C. for 12 hr. LCMS showed the
reaction was completed. The reaction mixture was diluted with water
(30 mL) and extracted with ethyl acetate (20 mL.times.2). The
combined organic phase was washed with saturated brine (20
mL.times.2), dried with anhydrous sodium sulfate, filtered and
concentrated in vacuum. The residue was purified by silica gel
chromatography (dichloromethane:methanol=1:0 to 50:1). Tert-butyl
4-[4-[3-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-
-1-yl]propoxy]phenyl]piperazine-1-carboxylate (480 mg, 0.73 mmol,
72% yield) was obtained as a yellow solid. LCMS:MS (ESI) m/z: 661.2
[M+1].sup.+. .sup.1H NMR: (400 MHz, CHLOROFORM-d) .delta. 8.26 (s,
1H), 7.70 (d, J=8.4 Hz, 1H), 7.29 (d, J=2.0 Hz, 1H), 7.07-7.05 (m,
1H), 6.91-6.81 (m, 4H), 4.94 (dd, J=12.0, 6.2 Hz, 1H), 4.02-3.99
(m, 2H), 3.59-3.56 (m, 4H), 3.45-3.43 (m, 4H), 3.02-3.00 (m, 4H),
2.87-2.86 (m, 4H), 2.62-2.59 (m, 4H), 2.15-2.05 (m, 1H), 2.01-1.98
(m, 2H), 1.56-1.54 (m, 1H), 1.48 (s, 9H). Chemical Formula:
C.sub.35H.sub.44N.sub.6O.sub.7, Molecular Weight: 660.76
9. Step--Synthesis of
2-(2,6-dioxopiperidin-3-yl)-5-(4-(3-(4-(piperazin-1-yl)phenoxy)propyl)pip-
erazin-1-yl)isoindoline-1,3-dione HCl Salt
##STR00768##
[1800] A mixture of tert-butyl
4-[4-[3-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-
-1-yl]propoxy]phenyl]piperazine-1-carboxylate (480 mg, 0.73 mmol, 1
eq) in hydrochloride/dioxane (8 mL, 4 M) was stirred at 25.degree.
C. for 1 hr. LCMS showed starting material was consumed completely
and one main peak with desired mass was detected. The mixture was
concentrated under reduced pressure. Crude compound
2-(2,6-dioxo-3-piperidyl)-5-[4-[3-(4-piperazin-1-ylphenoxy)propyl]piperaz-
in-1-yl]isoindoline-1,3-dione (500 mg, crude, hydrochloride) was
obtained as a yellow solid, which was confirmed by HNMR. LCMS: MS
(ESI) m/z: 561.3 [M+1]-. .sup.1H NMR: (400 MHz, MeOD) .delta. 7.80
(d, J=8.8 Hz, 1H), 7.51 (d, J=2.4 Hz, 1H), 7.39 (dd, J=8.4, 2.4 Hz,
1H), 7.12 (d, J=9.2 Hz, 2H), 6.99-6.97 (m, 2H), 5.14-5.09 (m, 1H),
4.22-4.15 (m, 2H), 4.14-4.13 (m, 4H), 3.75-3.73 (m, 4H), 3.48-3.45
(m, 1H), 3.44-3.41 (m, 8H), 3.40-3.26 (m, 1H), 2.79-2.78 (m, 1H),
2.75-2.76 (m, 2H), 2.35-2.34 (m, 2H), 2.33-2.12 (m, 1H). Chemical
Formula: C.sub.30H.sub.36N.sub.6O.sub.5, Molecular Weight:
560.64
10. Step--Synthesis of
2-(2,6-dioxo-3-piperidyl)-5-[4-[3-[4-[4-[2-[[1-isopropyl-6-[[2-(4-methoxy-
-1-piperidyl)pyrimidin-4-yl]amino]imidazo[4,5-c]pyridin-2-yl]methoxy]ethyl-
]piperazin-1-yl]phenoxy]propyl]piperazin-1-yl]isoindoline-1,3-dione
##STR00769##
[1802] To the mixture of
2-(2,6-dioxo-3-piperidyl)-5-[4-[3-(4-piperazin-1-ylphenoxy)propyl]piperaz-
in-1-yl]isoindoline-1,3-dione (136 mg, 0.23 mmol, 1 eq,
hydrochloride) and
2-[[1-isopropyl-6-[[2-(4-methoxy-1-piperidyl)pyrimidin-4-yl]amino]imidazo-
[4,5-c]pyridin-2-yl]methoxy]acetaldehyde (100 mg, 0.23 mmol, 1 eq)
in dichloroethane (5 mL) was added triethylamine (46 mg, 0.46 mmol,
2 eq). The mixture was stirred at 25.degree. C. for 2 hours. Then
sodium triacetoxy borohydride (96 mg, 0.46 mmol, 2 eq) was added to
the mixture and was stirred at 25.degree. C. for 10 hours. LCMS
showed formation of a new peak with the desired mass. The mixture
was concentrated under reduced pressure. The residue was purified
by prep-Thin-Layer chromatography (dichloromethane:methanol=10:1,
R.sub.t=0.15). Then it was purified by prep-HPLC [FA].
2-(2,6-dioxo-3-piperidyl)-5-[4-[3-[4-[4-[2-[[1-isopropyl-6-[[2-(4-methoxy-
-1-piperidyl)pyrimidin-4-yl]amino]imidazo[4,5-c]pyridin-2-yl]methoxy]ethyl-
]piperazin-1-yl]phenoxy]propyl]piperazin-1-yl]isoindoline-1,3-dione
(13.7 mg, 0.01 mmol, 5% yield, 93% purity) was obtained as a yellow
solid, which was confirmed by HNMR and LCMS. LCMS: MS (ESI) m/z:
984.4 [M+1].sup.+. .sup.1H NMR: (400 MHz, MeOD) .delta. 8.67 (s,
1H), 8.50 (s, 1H), 7.96 (d, J=6.0 Hz, 1H), 7.73 (d, J=8.8 Hz, 1H),
7.43 (d, J=2.0 Hz, 1H), 7.30 (d, J=8.4, 2.4 Hz, 1H), 6.98 (d, J=9.2
Hz, 2H), 6.89 (d, J=8.8 Hz, 2H), 6.42-6.38 (m, 1H), 5.10-5.08 (m,
1H), 4.94 (s, 2H), 4.31-4.26 (m, 2H), 4.07-4.04 (m, 2H), 3.94-3.85
(m, 2H), 3.60-3.58 (m, 4H), 3.43-3.42 (m, 1H), 3.42 (s, 3H),
3.25-3.24 (m, 4H), 3.23-3.20 (m, 6H), 2.96-2.95 (m, 4H), 2.89-2.87
(m, 3H), 2.82-2.75 (m, 2H), 2.12-2.10 (m, 3H), 2.05-1.95 (m, 2H),
1.73 (d, J=6.8 Hz, 6H), 1.61-1.51 (m, 2H). Chemical Formula:
C.sub.52H.sub.65N.sub.13O.sub.7, Molecular Weight: 984.16
Synthesis of Example 302
(2S,4R)-4-hydroxy-1-((S)-2-(2-((1r,3s)-3-(4-(4-(2-((1-isopropyl-6-((2-(4-m-
ethoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)m-
ethoxy)ethyl)piperazin-1-yl)phenoxy)cyclobutoxy)acetamido)-3,3-dimethylbut-
anoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
##STR00770##
[1804] Synthetic Route:
##STR00771## ##STR00772##
1. Step--Synthesis of tert-butyl
4-(4-((1r,3r)-3-(benzyloxy)cyclobutoxy)phenyl)piperazine-1-carboxylate
##STR00773##
[1806] To a solution of tert-butyl
4-(4-hydroxyphenyl)piperazine-1-carboxylate (1.5 g, 5.39 mmol, 1
eq) and cis-3-benzyloxycyclobutanol (1.44 g, 8.08 mmol, 1.5 eq) in
toluene (15 mL) was added 1,1'-(azodicarbonyl)dipiperidine (2.72 g,
10.78 mmol, 2 eq) and tributylphosphine (2.18 g, 10.78 mmol, 2.66
mL, 2 eq). The mixture was stirred at 100.degree. C. for 12 hours.
LCMS showed the reaction was completed. The reaction mixture was
diluted with water (50 mL) and extracted with ethyl acetate (30
mL.times.2). The combined organic phase was washed with saturated
brine (30 mL.times.2), dried with anhydrous sodium sulfate,
filtered and concentrated in vacuum. The residue was purified by
preparative High Performance Liquid Chromatography (Formic acid
buffered eluent) to afford tert-butyl
4-(4-((1r,3r)-3-(benzyloxy)cyclobutoxy)phenyl)piperazine-1-carboxylate
(750 mg, 1.71 mmol, 31% yield) as a brown solid. LCMS: MS (ESI)
m/z: 439.2 [M+1].sup.+. .sup.1H NMR: (400 MHz, CDCl.sub.3) .delta.:
7.28-7.36 (m, 5H), 6.89 (d, J=7.2 Hz, 2H), 6.73 (d, J=7.2 Hz, 2H),
4.80-4.83 (m, 1H), 4.45 (s, 2H), 4.32-4.35 (m, 1H), 3.57-3.60 (m,
4H), 3.00-3.03 (m, 4H), 2.42-2.49 (m, 4H), 1.49 (s, 9H). Chemical
Formula: C.sub.26H.sub.34N.sub.2O4, Molecular Weight: 438.56
2. Step--Synthesis of tert-butyl
4-(4-((1r,3r)-3-hydroxycyclobutoxy)phenyl)piperazine-1-carboxylate
##STR00774##
[1808] To a solution of tert-butyl
4-(4-((1r,3r)-3-(benzyloxy)cyclobutoxy)phenyl)piperazine-1-carboxylate
(0.9 g, 2.05 mmol, 1 eq) in methanol (20 mL) and tetrahydrofuran
(20 mL) was added palladium on activated carbon catalyst (0.2 g,
10% purity), the mixture was degassed and purged with hydrogen
several times, then stirred at 25.degree. C. for 12 hours under
hydrogen (15 psi) atmosphere. The desired MS was observed by LCMS,
thin layer chromatography (petroleum ether/ethyl acetate=3/1)
showed one major new spot was detected. The mixture was filtered;
the filtrate was concentrated under reduced pressure. The residue
was purified by silica gel chromatography (petroleum ether/ethyl
acetate=10/1 to 3/1) to afford tert-butyl
4-(4-((1r,3r)-3-hydroxycyclobutoxy)phenyl)piperazine-1-carboxylate
(0.62 g, 1.78 mmol, 86% yield) as a light yellow solid. LCMS: MS
(ESI) m/z: 349.1 [M+1].sup.+. .sup.1H NMR: (400 MHz, CDCl.sub.3)
.delta.: 6.86-6.91 (m, 2H), 6.71-6.76 (m, 2H), 4.80-4.88 (m, 1H),
4.58-4.69 (m, 1H), 3.54-3.62 (m, 4H), 2.97-3.05 (m, 4H), 2.46-2.55
(m, 2H), 2.34-2.44 (m, 2H), 1.49 (s, 9H). Chemical Formula:
C.sub.19H.sub.28N.sub.2O.sub.4, Molecular Weight: 348.44
3. Step--Synthesis of tert-butyl
4-(4-((1r,3r)-3-(2-ethoxy-2-oxoethoxy)cyclobutoxy)phenyl)piperazine-1-car-
boxylate
##STR00775##
[1810] To a solution of tert-butyl
4-(4-((1r,3r)-3-hydroxycyclobutoxy)phenyl)piperazine-1-carboxylate
(0.56 g, 1.61 mmol, 1 eq) and ethyl 2-diazoacetate (733 mg, 6.43
mmol, 4 eq) in dichloromethane (20 mL) was added rhodium(II)
acetate (18 mg, 0.08 mmol, 0.05 eq), the mixture was stirred at
25.degree. C. for 2 hours. Thin layer chromatography (petroleum
ether/ethyl acetate=3/1) showed the starting material was consumed
completely and one major new spot was detected. The mixture was
concentrated under reduced pressure. The residue was purified by
silica gel chromatography (petroleum ether/ethyl acetate=5/1) to
afford tert-butyl
4-(4-((1r,3r)-3-(2-ethoxy-2-oxoethoxy)cyclobutoxy)phenyl)piperazine-1-car-
boxylate (0.42 g, 0.96 mmol, 60% yield) as a light yellow solid.
.sup.1H NMR: (400 MHz, CDCl.sub.3) .delta.: 6.85-6.92 (m, 2H),
6.67-6.76 (m, 2H), 4.76-4.85 (m, 1H), 4.32-4.37 (m, 1H), 4.24 (q,
J=7.2 Hz, 1H), 4.02 (s, 2H), 3.57 (t, J=4.8 Hz, 4H), 3.00 (t, J=4.8
Hz, 4H), 2.40-2.56 (m, 4H), 1.49 (s, 9H), 1.30 (t, J=7.2 Hz, 3H).
Chemical Formula: C.sub.23H.sub.34N.sub.2O.sub.6, Molecular Weight:
434.53
4. Step--Synthesis of
2-((1r,3r)-3-(4-(4-(tert-butoxycarbonyl)piperazin-1-yl)phenoxy)cyclobutox-
y)acetic acid
##STR00776##
[1812] To a solution of tert-butyl
4-(4-((1r,3r)-3-(2-ethoxy-2-oxoethoxy)cyclobutoxy)phenyl)piperazine-1-car-
boxylate (0.42 g, 0.96 mmol, 1 eq) in tetrahydrofuran (10 mL) and
water (1 mL) was added lithium hydroxide (69 mg, 2.90 mmol, 3 eq),
the mixture was stirred at 50.degree. C. for 2 hours. Thin layer
chromatography (petroleum ether/ethyl acetate=3/1) showed the
starting material was consumed completely and the desired MS was
detected by LCMS. The mixture was concentrated under reduced
pressure, the residue was diluted with water (20 mL), the pH of
aqueous phase was adjusted to 6 by hydrochloric acid (1 M), then
extracted with ethyl acetate (20 mL.times.3), the organic layer was
dried over anhydrous sodium sulfate, filtered and concentrated
under reduced pressure to afford
2-((1r,3r)-3-(4-(4-(tert-butoxycarbonyl)piperazin-1-yl)phenoxy)cyclobutox-
y)acetic acid (0.32 g, 0.79 mmol, 81% yield) as a light yellow oil.
LCMS: MS (ESI) m/z: 407.1 [M+1].sup.+. .sup.1H NMR: (400 MHz,
CDCl.sub.3) .delta.: 6.89 (d, J=8.8 Hz, 2H), 6.71 (d, J=8.8 Hz,
2H), 4.69-4.79 (m, 1H), 4.19-4.29 (m, 1H), 3.94 (s, 2H), 3.41-3.45
(m, 4H), 2.74-3.00 (m, 4H), 2.37-2.42 (m, 2H), 2.21-2.29 (m, 2H),
1.41 (s, 9H). Chemical Formula: C.sub.21H.sub.30N.sub.2O.sub.6,
Molecular Weight: 406.47
5. Step--Synthesis of tert-butyl
4-[4-[(1s,3r)-3-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-y-
l)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino-
]-2-oxo-ethoxy]cyclobutoxy]phenyl]piperazine-1-carboxylate
##STR00777##
[1814] To a solution of
2-[(1r,3r)-3-[4-(4-tert-butoxycarbonylpiperazin-1-yl)phenoxy]cyclobutoxy]-
acetic acid (0.31 g, 0.76 mmol, 1 eq) and
(2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[4-(4-methylt-
hiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (328 mg, 0.76
mmol, 1 eq) in N,N-dimethylformamide (5 mL) was added
1-hydroxybenzotriazole (154 mg, 1.14 mmol, 1.5 eq),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (219
mg, 1.14 mmol, 1.5 eq) and N,N-diisopropylethylamine (295 mg, 2.29
mmol, 0.4 mL, 3 eq), the mixture was stirred at 25.degree. C. for 4
hours. The desired MS was observed by LCMS. The mixture was diluted
with water (50 mL), and then extracted with ethyl acetate (30
mL.times.3), the organic layer was washed with brine (50
mL.times.2), dried over anhydrous sodium sulfate, filtered and
concentrated under reduced pressure. The residue was purified by
silica gel chromatography (ethyl acetate/methanol=10/1) to afford
tert-butyl
4-[4-[(1s,3r)-3-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-y-
l)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino-
]-2-oxo-ethoxy]cyclobutoxy]phenyl]piperazine-1-carboxylate (0.4 g,
0.49 mmol, 64% yield) as a light yellow oil. LCMS: MS (ESI) m/z:
819.3 [M+1]+. .sup.1H NMR: (400 MHz, CDCl.sub.3) .delta.: 8.68 (s,
1H), 7.31-7.40 (m, 5H), 6.87 (d, J=8.8 Hz, 2H), 6.72 (d, J=8.8 Hz,
2H), 4.71-4.83 (m, 2H), 4.52-4.56 (m, 2H), 4.24-4.38 (m, 2H),
4.07-4.12 (m, 1H), 3.80-3.90 (m, 2H), 3.56 (t, J=5.2 Hz, 4H), 2.99
(t, J=5.2 Hz, 4H), 2.55-2.59 (m, 1H), 2.52 (s, 3H), 2.42-2.47 (m,
4H), 2.08-2.17 (m, 2H), 2.05 (d, J=4.8 Hz, 1H), 1.48 (s, 9H), 0.95
(s, 9H). Chemical Formula: C.sub.43H.sub.58N.sub.6O.sub.8S,
Molecular Weight: 819.02
6. Step--Synthesis of
(2S,4R)-1-((S)-3,3-dimethyl-2-(2-((1r,3s)-3-(4-(piperazin-1-yl)phenoxy)cy-
clobutoxy)acetamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-
pyrrolidine-2-carboxamide HCl Salt
##STR00778##
[1816] A solution of tert-butyl
4-[4-[(1s,3r)-3-[2-[[(S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-yl-
)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-
-2-oxo-ethoxy]cyclobutoxy]phenyl]piperazine-1-carboxylate (0.4 g,
0.49 mmol, 1 eq) in hydrochloric acid/dioxane (4 M, 5 mL) was
stirred at 25.degree. C. for 1 hour. The desired MS was observed by
LCMS. The mixture was concentrated under reduced pressure. The
residue was purified by preparative High Performance Liquid
Chromatography (column: Phenomenex Synergi C18 150*25*10 um; mobile
phase: [water (0.05% HCl)-ACN]; B %: 15%-42%, 9 min) to afford
(2S,4R)-1-((S)-3,3-dimethyl-2-(2-((1r,3s)-3-(4-(piperazin-1-yl)phenoxy)cy-
clobutoxy)acetamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-
pyrrolidine-2-carboxamide (200 mg, 0.26 mmol, 54% yield,
hydrochloride) as a colorless oil. LCMS: MS (ESI) m/z: 719.1
[M+1].sup.+. .sup.1H NMR: (400 MHz, CDCl.sub.3) .delta.: 9.01 (s,
3H), 8.61 (s, 1H), 7.37-7.48 (m, 5H), 6.93 (d, J=7.8 Hz, 2H), 6.75
(d, J=8.8 Hz, 2H), 4.73-4.82 (m, 1H), 4.56 (d, J=9.6 Hz, 1H),
4.33-4.49 (m, 3H), 4.24-4.29 (m, 2H), 3.85-3.92 (m, 2H), 3.59-3.65
(m, 4H), 3.16-3.25 (m, 8H), 2.44 (s, 3H), 2.22-2.30 (m, 2H),
2.03-2.10 (m, 1H), 1.85-1.92 (m, 1H), 0.94 (s, 9H). Chemical
Formula: C.sub.38H.sub.51ClN.sub.6O.sub.6S, Molecular Weight:
755.37
7. Step--Synthesis of
(2S,4R)-4-hydroxy-1-((S)-2-(2-((1r,3s)-3-(4-(4-(2-((1-isopropyl-6-((2-(4--
methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)-
methoxy)ethyl)piperazin-1-yl)phenoxy)cyclobutoxy)acetamido)-3,3-dimethylbu-
tanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
##STR00779##
[1818] To a solution of
(2S,4R)-1-((S)-3,3-dimethyl-2-(2-((1r,3s)-3-(4-(piperazin-1-yl)phenoxy)cy-
clobutoxy)acetamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-
pyrrolidine-2-carboxamide (50 mg, 0.07 mmol, 1 eq, hydrochloride)
in methanol (1 mL) and dichloromethane (1 mL) was added sodium
acetate (22 mg, 0.26 mmol, 4 eq), the mixture was stirred at
25.degree. C. for 0.5 hour, then
2-[[1-isopropyl-6-[[2-(4-methoxy-1-piperidyl)pyrimidin-4-yl]amino]imidazo-
[4,5-c] pyridin-2-yl]methoxy]acetaldehyde (29 mg, 0.07 mmol, 1 eq)
was added, the mixture was stirred at 25.degree. C. for 1 hour,
then sodium cyanoborohydride (8 mg, 0.13 mmol, 2 eq) was added, the
mixture was stirred at 25.degree. C. for 0.5 hour. The desired MS
was observed by LCMS. The mixture was concentrated under reduced
pressure. The residue was purified by preparative High Performance
Liquid Chromatography (column: Boston Green ODS 150*30 5 u; mobile
phase: [water (0.225% FA)-ACN]; B %: 18%-45%,10 min) to afford
(2S,4R)-4-hydroxy-1-((S)-2-(2-((1r,3s)-3-(4-(4-(2-((1-isopropyl-6-((2-(4--
methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-imidazo[4,5-c]pyridin-2-yl)-
methoxy)ethyl)piperazin-1-yl)phenoxy)cyclobutoxy)acetamido)-3,3-dimethylbu-
tanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
(29.6 mg, 0.02 mmol, 37% yield, 98.8% purity, formate) as a
off-white solid. LCMS: MS (ESI) m/z: 1142.6 [M+1].sup.+. .sup.1H
NMR: (400 MHz, MeOD) .delta.: 8.84 (m, 1H), 8.64 (s, 1H), 8.47 (s,
1H), 8.37 (brs, 1H), 7.94 (d, J=6.0 Hz, 1H), 7.38-7.48 (m, 4H),
6.88-6.94 (m, 2H), 6.70-6.76 (m, 2H), 6.36 (d, J=6.0 Hz, 1H),
4.90-4.95 (m, 3H), 4.70-4.82 (m, 2H), 4.46-4.62 (m, 3H), 4.21-4.39
(m, 4H), 3.79-4.02 (m, 6H), 3.43-3.57 (m, 3H), 3.38 (s, 3H),
3.08-3.20 (m, 9H), 2.66 (s, 1H), 2.43-2.55 (m, 5H), 2.32-2.41 (m,
2H), 2.18-2.28 (m, 1H), 2.04-2.15 (m, 1H), 1.93-2.01 (m, 2H), 1.69
(d, J=6.8 Hz, 6H), 1.51-1.62 (m, 2H), 1.04 (m, 9H). Chemical
Formula: C.sub.60H.sub.79N.sub.13O.sub.8S, Molecular Weight:
1142.42
Synthesis of Example 307
2-(7-((4-(3-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)pi-
perazin-1-yl)piperidin-1-yl)propoxy)butyl)amino)-1-oxoisoindolin-2-yl)-2-(-
5-fluoro-2-hydroxyphenyl)-N-(thiazol-2-yl)acetamide
##STR00780##
[1820] Synthetic Route:
##STR00781## ##STR00782## ##STR00783## ##STR00784##
1. Step--Synthesis of methyl 2-(bromomethyl)-6-nitro-benzoate
##STR00785##
[1822] To a solution of methyl 2-methyl-6-nitro-benzoate (2.6 g,
13.32 mmol, 1 eq) in carbon tetrachloride (25 mL) was added benzoyl
peroxide (322 mg, 1.33 mmol, 0.1 eq) and NBS (2.49 g, 13.99 mmol,
1.05 eq). The mixture was stirred at 80.degree. C. for 10 hours.
Thin layer chromatography (Petroleum ether/Ethyl acetate=10/1)
showed the starting material was consumed completely. The reaction
mixture was concentrated under reduced pressure. The residue was
diluted with water (20 mL), then extracted with ethyl acetate (50
mL.times.3). The combined organic layers were washed with brine
(100 mL.times.3), dried over anhydrous sodium sulfate, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by silica gel chromatography (Petroleum ether/Ethyl
acetate=30/1) to give the product methyl
2-(bromomethyl)-6-nitro-benzoate (2.2 g, 8.03 mmol, 60% yield) as a
white solid. .sup.1H NMR: (400 MHz, CDCl.sub.3) .delta.: 8.08 (d,
J=8.0 Hz, 1H), 7.77 (d, J=8.0 Hz, 1H), 7.60 (t, J=8.0 Hz, 1H), 4.58
(s, 2H), 3.99 (s, 3H). Chemical Formula: C.sub.9H.sub.8BrNO.sub.4,
Molecular Weight: 274.07
2. Step--Synthesis of 4-(3-benzyloxypropoxy)butan-1-ol
##STR00786##
[1824] A mixture of butane-1,4-diol (2.36 g, 26.19 mmol, 2.3 mL,
1.2 eq), sodium hydride (960 mg, 24.01 mmol, 60% in mineral oil,
1.1 eq), in tetrahydrofuran (50 mL) was degassed and purged with
nitrogen for 3 times and stirred at 25.degree. C. for 2 hours, then
to this mixture was added 3-bromopropoxymethylbenzene (5 g, 21.82
mmol, 3.8 mL, 1 eq) and stirred at 70.degree. C. for 10 hours under
nitrogen atmosphere. Thin layer chromatography (Petroleum
ether/Ethyl acetate=3/1) indicated the starting material was
consumed completely and one new spot was formed. The reaction
mixture was concentrated under reduced pressure. The residue was
diluted with water (40 mL), then extracted with ethyl acetate (100
mL.times.3). The combined organic layers were washed with brine
(200 mL.times.3), dried over anhydrous sodium sulfate, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by silica gel chromatography (Petroleum ether/Ethyl
acetate=5/1) to give 4-(3-benzyloxypropoxy)butan-1-ol (1.2 g, 5.04
mmol, 23% yield) as a colorless oil. .sup.1H NMR: (400 MHz,
CDCl.sub.3) .delta.: 7.30-7.40 (m, 5H), 4.51 (s, 2H), 3.64 (t,
J=5.6 Hz, 2H), 3.54-3.57 (m, 4H), 3.47 (t, J=5.6 Hz, 2H), 1.88-1.91
(m, 2H), 1.65-1.70 (m, 4H). Chemical Formula:
C.sub.14H.sub.22O.sub.3, Molecular Weight: 238.32
3. Step--Synthesis of 4-(3-benzyloxypropoxy)butanal
##STR00787##
[1826] To a solution of 4-(3-benzyloxypropoxy)butan-1-ol (1.2 g,
5.04 mmol, 1 eq) in dichloromethane (50 mL) was added
1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (3.20 g,
7.55 mmol, 1.5 eq). The mixture was stirred at 25.degree. C. for 1
hour. Thin layer chromatography (Petroleum ether/Ethyl acetate=3/1)
showed the starting material was consumed completely and one new
spot was detected. The mixture was filtered, the filtrate was
concentrated under reduced pressure to give a residue. The residue
was purified by silica gel chromatography (Petroleum ether/Ethyl
acetate=5/1) to give 4-(3-benzyloxypropoxy)butanal (0.9 g, 3.81
mmol, 75% yield) as a yellow oil. .sup.1H NMR: (400 MHz,
CDCl.sub.3) .delta.: 9.76 (s, 1H), 7.28-7.38 (m, 5H), 4.51 (s, 2H),
3.53 (dt, J=16.4, 6.4 Hz, 4H), 3.44 (t, J=6.4 Hz, 2H), 2.47-2.51
(m, 2H), 1.81-1.94 (m, 4H). Chemical Formula:
C.sub.14H.sub.20O.sub.3, Molecular Weight: 236.31
4. Step--Synthesis of 2-(benzyloxy)-5-fluorobenzaldehyde
##STR00788##
[1828] To a solution of 5-fluoro-2-hydroxy-benzaldehyde (25 g,
178.43 mmol, 1 eq) in N,N-dimethylformamide (250 mL) was added
potassium carbonate (49.32 g, 356.86 mmol, 2 eq) and
bromomethylbenzene (36.62 g, 214.12 mmol, 25.4 mL, 1.2 eq). The
mixture was stirred at 50.degree. C. for 2 hours. Thin layer
chromatography (Petroleum ether/Ethyl acetate=5/1) indicated the
starting material was consumed completely and one new spot was
formed. The reaction mixture was concentrated under reduced
pressure. The residue was diluted with water (500 mL), then
extracted with ethyl acetate (400 mL.times.3). The combined organic
layers were washed with brine (500 mL.times.3), dried over
anhydrous sodium sulfate, filtered and concentrated under reduced
pressure to give a residue. The residue was purified by silica gel
chromatography (Petroleum ether/Ethyl acetate=5/1) to give the
product 2-benzyloxy-5-fluoro-benzaldehyde (40 g, 173.74 mmol, 97%
yield) as a white solid. .sup.1H NMR: (400 MHz, DMSO-d.sub.6)
.delta.: 10.37 (s, 1H), 7.49-7.57 (m, 3H), 7.35-7.44 (m, 5H), 5.29
(s, 2H). Chemical Formula: C.sub.14H.sub.11FO.sub.2, Molecular
Weight: 230.07
5. Step--Synthesis of
2-amino-2-(2-benzyloxy-5-fluoro-phenyl)acetonitrile
##STR00789##
[1830] To a solution of ammonium chloride (18.59 g, 347.47 mmol, 2
eq) in methanol (200 mL) was added trimethylsilyl cyanide (25.85 g,
260.60 mmol, 32.6 mL, 1.5 eq) and ammonium hydroxide (1.95 mol, 300
mL, 25% purity, 11.21 eq), after 30 min then this solution was
added to a solution of 2-benzyloxy-5-fluoro-benzaldehyde (40 g,
173.74 mmol, 1 eq) in methanol (400 mL). The mixture was stirred at
25.degree. C. for 20 hours. Thin layer chromatography (Petroleum
ether/Ethyl acetate=5/1) showed the starting material was consumed
completely and one new spot was detected. The reaction mixture was
concentrated under reduced pressure. The residue was diluted with
water (200 mL), then extracted with ethyl acetate (400 mL.times.3).
The combined organic layers were washed with brine (300
mL.times.3), dried over anhydrous sodium sulfate, filtered and
concentrated under reduced pressure to give a residue
2-amino-2-(2-benzyloxy-5-fluoro-phenyl)acetonitrile (50 g, crude)
as a black oil. .sup.1H NMR: (400 MHz, DMSO-d.sub.6) .delta.: 7.52
(d, J=7.2 Hz, 2H), 7.31-7.43 (m, 4H), 7.13-7.20 (m, 2H), 5.20 (s,
2H), 5.08 (s, 1H), 2.89 (brs, 2H). Chemical Formula:
C.sub.15H.sub.13N.sub.2FO, Molecular Weight: 256.27
6. Step--Synthesis of methyl
2-amino-2-(2-benzyloxy-5-fluoro-phenyl)acetate
##STR00790##
[1832] A solution of
2-amino-2-(2-benzyloxy-5-fluoro-phenyl)acetonitrile (50 g, 195.10
mmol, 1 eq) in hydrochloric acid/methanol (4 M, 500 mL) was stirred
at 25.degree. C. for 10 hours. LCMS showed the starting material
was consumed completely and one main peak with desired m/z was
detected. The reaction mixture was concentrated under reduced
pressure. The residue was diluted with water (400 mL), the pH of
the aqueous phase was adjusted to 7-8 by ammonium hydroxide (25%),
then extracted with ethyl acetate (500 mL.times.3). The combined
organic layers were washed with brine (400 mL.times.3), dried over
anhydrous sodium sulfate, filtered and concentrated under reduced
pressure to give a residue. The residue was purified by silica gel
chromatography (Petroleum ether/Ethyl acetate=5/1) to give the
product methyl 2-amino-2-(2-benzyloxy-5-fluoro-phenyl)acetate (45
g, 155.55 mmol, 79% yield) as a brown oil. .sup.1H NMR: (400 MHz,
DMSO-d.sub.6) .delta.: 7.38-7.46 (m, 4H), 7.31-7.36 (m, 1H),
7.22-7.27 (m, 1H), 7.05-7.10 (m, 2H), 5.10 (s, 2H), 4.71 (s, 1H),
3.52 (s, 3H), 3.34 (brs, 2H). Chemical Formula:
C.sub.16H.sub.16FNO.sub.3, Molecular Weight: 289.30.
7. Step--Synthesis of methyl
2-(2-benzyloxy-5-fluoro-phenyl)-2-(7-nitro-1-oxo-isoindolin-2-yl)acetate
##STR00791##
[1834] To a solution of methyl
2-amino-2-(2-benzyloxy-5-fluoro-phenyl)acetate (3.2 g, 11.06 mmol,
1 eq) in N,N-dimethylformamide (20 mL) was added
diisopropylethylamine (2.86 g, 22.12 mmol, 3.8 mL, 2.0 eq) and
methyl 2-(bromomethyl)-6-nitro-benzoate (3.03 g, 11.06 mmol, 1 eq),
the mixture was stirred at 50.degree. C. for 5 hours. And then the
mixture was stirred at 100.degree. C. for 10 hours. LCMS showed the
starting material was consumed completely and one main peak with
desired m/z was detected. The reaction mixture was concentrated
under reduced pressure. The residue was diluted with water (20 mL),
then extracted with ethyl acetate (50 mL.times.3). The combined
organic layers were washed with brine (50 mL.times.3), dried over
anhydrous sodium sulfate, filtered and concentrated under reduced
pressure to give a residue. The residue was purified by silica gel
chromatography (Petroleum ether/Ethyl acetate=3/1) to afford methyl
2-(2-benzyloxy-5-fluoro-phenyl)-2-(7-nitro-1-oxo-isoindolin-2-yl)
acetate (4 g, 8.88 mmol, 80% yield) as a white solid. LCMS: MS
(ESI) m/z: 451.1 [M+1].sup.+ 1H NMR: (400 MHz, DMSO-d.sub.6)
.delta.: 7.91-7.95 (m, 1H), 7.81-7.87 (m, 2H), 7.36-7.41 (m, 2H),
7.26-7.34 (m, 5H), 7.23 (d, J=8.4 Hz, 1H), 6.20 (s, 1H), 5.17 (q,
J=12.0 Hz, 2H), 4.64 (d, J=18.0 Hz, 1H), 4.12 (d, J=18.0 Hz, 1H),
3.60 (s, 3H). Chemical Formula: C.sub.24H.sub.19FN.sub.2O.sub.6,
Molecular Weight: 450.42
8. Step--Synthesis of methyl
2-(7-amino-1-oxo-isoindolin-2-yl)-2-(5-fluoro-2-hydroxy-phenyl)acetate
##STR00792##
[1836] To a solution of methyl
2-(2-benzyloxy-5-fluoro-phenyl)-2-(7-nitro-1-oxo-isoindolin-2-yl)acetate
(4 g, 8.88 mmol, 1 eq) in tetrahydrofuran (150 mL) was added
palladium on activated carbon catalyst (0.5 g, 10% purity) and
purged with hydrogen for 3 times, and then the mixture was stirred
at 25.degree. C. for 10 hours under hydrogen (15 Psi). LCMS showed
the starting material was consumed completely and one main peak
with desired m/z was detected. The reaction mixture was filtered by
diatomite and concentrated under reduced pressure to give methyl
2-(7-amino-1-oxo-isoindolin-2-yl)-2-(5-fluoro-2-hydroxy-phenyl)
acetate (3.8 g, crude) as a yellow oil, which was used into the
next step without further purification. LCMS: MS (ESI) m/z: 331.2
[M+1].sup.+. .sup.1H NMR: EW8426-36-P1A, (400 MHz, DMSO-d.sub.6)
.delta.: 9.99 (s, 1H), 7.21 (t, J=8.0 Hz, 1H), 6.98-7.10 (m, 2H),
6.83-6.90 (m, 1H), 6.59 (t, J=8.0 Hz, 2H), 6.04-6.07 (m, 3H), 4.49
(d, J=17.6 Hz, 1H), 3.87 (d, J=17.6 Hz, 1H), 3.70 (s, 3H). Chemical
Formula: C.sub.17H.sub.15FN.sub.2O.sub.4, Molecular Weight:
330.31.
9. Step--Synthesis of methyl
2-[7-[4-(3-benzyloxypropoxy)butylamino]-1-oxo-isoindolin-2-yl]-2-(5-fluor-
o-2-hydroxy-phenyl)acetate
##STR00793##
[1838] To a solution of methyl
2-(7-amino-1-oxo-isoindolin-2-yl)-2-(5-fluoro-2-hydroxy-phenyl)acetate
(1.40 g, 4.23 mmol, 1 eq) in acetic acid (2 mL) and methanol (20
mL) was added 4-(3-benzyloxypropoxy)butanal (1.0 g, 4.23 mmol, 1
eq), the mixture was stirred at 25.degree. C. for 1 hour. Then
borane; 2-methylpyridine (905 mg, 8.46 mmol, 2 eq) was added, the
mixture was stirred at 25.degree. C. for 1 hour. LCMS showed one
main peak with desired m/z was detected. The mixture was
concentrated under reduced pressure. The residue was diluted with
water (20 mL), then extracted with ethyl acetate (50 mL.times.3).
The combined organic layers were washed with brine (50 mL.times.3),
dried over anhydrous sodium sulfate, filtered and concentrated
under reduced pressure to give a residue. The residue was purified
by preparative High Performance Liquid Chromatography (Formic acid
condition) to give methyl
2-[7-[4-(3-benzyloxypropoxy)butylamino]-1-oxo-isoindolin-2-yl]-2-(5-fluor-
o-2-hydroxy-phenyl) acetate (1.5 g, 2.72 mmol, 64% yield) as a
yellow oil. LCMS: MS (ESI) m/z: 551.2 [M+1].sup.+ .sup.1H NMR: (400
MHz, CDCl.sub.3) .delta.: 10.00 (s, 1H), 7.23-7.37 (m, 6H),
6.99-7.10 (m, 2H), 6.88 (dd, J=8.8, 4.8 Hz, 1H), 6.52-6.66 (m, 3H),
6.04 (s, 1H), 4.50 (d, J=17.6 Hz, 1H), 4.43 (s, 2H), 3.88 (d,
J=17.6 Hz, 1H), 3.68 (s, 3H), 3.36-3.49 (m, 6H), 3.06-3.25 (m, 2H),
1.74-1.77 (m, 2H), 1.51-1.63 (m, 4H). Chemical Formula:
C.sub.31H.sub.35FN.sub.2O.sub.6, Molecular Weight: 550.62.
10. Step--Synthesis of methyl
2-[7-[4-(3-benzyloxypropoxy)butylamino]-1-oxo-isoindolin-2-yl]-2-(5-fluor-
o-2-tetrahydropyran-2-yloxy-phenyl)acetate
##STR00794##
[1840] To a solution of methyl
2-[7-[4-(3-benzyloxypropoxy)butylamino]-1-oxo-isoindolin-2-yl]-2-(5-fluor-
o-2-hydroxy-phenyl)acetate (1.2 g, 2.18 mmol, 1 eq) in
tetrahydrofuran (12 mL) was added 3,4-dihydro-2H-pyran (366 mg,
4.36 mmol, 0.4 mL, 2 eq) and p-toluenesulfonic acid (41 mg, 0.22
mmol, 0.1 eq), the mixture was stirred at 25.degree. C. for 4
hours. Thin Layer Chromatography (petroleum ether/ethyl
acetate=2/1) showed the starting material remained; one major new
spot with lower polarity was detected. The mixture was concentrated
under reduced pressure. The residue was purified by silica gel
chromatography (petroleum ether/ethyl acetate=10/1 to 3/1) to
afford methyl
2-[7-[4-(3-benzyloxypropoxy)butylamino]-1-oxo-isoindolin-2-yl]-2-(5-fluor-
o-2-tetrahydropyran-2-yloxy-phenyl)acetate (0.8 g, 1.26 mmol, 57%
yield) as a light yellow oil. .sup.1H NMR: (400 MHz, CDCl.sub.3)
.delta.: 7.28-7.34 (m, 4H), 7.11-7.25 (m, 2H), 6.91-7.08 (m, 2H),
6.43-6.74 (m, 3H), 6.31-6.36 (m, 1H), 5.19-5.55 (m, 1H), 4.88-4.96
(m, 1H), 4.57-4.73 (m, 1H), 4.51 (s, 2H), 4.01-4.05 (m, 1H),
3.92-3.95 (m, 1H), 3.76-3.80 (m, 3H), 3.51-3.59 (m, 6H), 3.20-3.29
(m, 2H), 1.86-1.91 (m, 2H), 1.68-1.75 (m, 4H), 1.60-1.65 (m, 4H),
1.53-1.57 (m, 2H). Chemical Formula:
C.sub.36H.sub.43FN.sub.2O.sub.7, Molecular Weight: 634.73.
11. Step--Synthesis of methyl
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[7-[4-(3-hydroxypropoxy)b-
utylamino]-1-oxo-isoindolin-2-yl]acetate
##STR00795##
[1842] A mixture of methyl
2-[7-[4-(3-benzyloxypropoxy)butylamino]-1-oxo-isoindolin-2-yl]-2-(5-fluor-
o-2-tetrahydropyran-2-yloxyphenyl) acetate (0.8 g, 1.26 mmol, 1
eq), palladium on activated carbon catalyst (0.1 g, 10% purity) in
tetrahydrofuran (10 mL) was degassed and purged with hydrogen for 3
times, and then the mixture was stirred at 25.degree. C. for 3
hours under hydrogen (15 Psi) atmosphere. Thin layer chromatography
(Petroleum ether/Ethyl acetate=1/1) showed the starting material
was consumed completely and one new spot was detected. The mixture
was filtered, the filtrate was concentrated under reduced pressure
to give methyl
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[7-[4-(3-hydroxypropoxy)b-
utylamino]-1-oxo-isoindolin-2-yl]acetate (0.5 g, 0.92 mmol, 72%
yield) as a yellow oil. .sup.1H NMR: (400 MHz, CDCl.sub.3) .delta.:
7.28-7.31 (m, 1H), 7.12-7.23 (m, 1H), 6.97-7.03 (m, 2H), 6.64 (s,
1H), 6.48-6.57 (m, 2H), 6.32 (s, 1H), 5.23-5.51 (m, 1H), 4.54-4.71
(m, 1H), 3.92-4.01 (m, 1H), 3.76-3.81 (m, 5H), 3.66-3.72 (m, 1H),
3.61-3.64 (m, 2H), 3.45-3.57 (m, 3H), 3.24-3.26 (m, 2H), 1.82-1.94
(m, 3H), 1.61-1.80 (m, 9H). Chemical Formula:
C.sub.29H.sub.37FN.sub.2O.sub.7, Molecular Weight: 544.61
12. Step--Synthesis of
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[7-[4-(3-hydroxypropoxy)b-
utylamino]-1-oxo-isoindolin-2-yl]acetic acid
##STR00796##
[1844] To a solution of methyl
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[7-[4-(3-hydroxypropoxy)b-
utylamino]-1-oxoisoindolin-2-yl]acetate (0.5 g, 0.92 mmol, 1 eq) in
tetrahydrofuran (10 mL) and water (2 mL) was added lithium
hydroxide (24 mg, 1.01 mmol, 1.1 eq). The mixture was stirred at
25.degree. C. for 1 hour. LCMS showed the starting material was
consumed completely and one peak with desired mass was detected.
The reaction mixture was concentrated under reduced pressure. The
residue was diluted with water (10 mL), the pH of the aqueous phase
was adjusted to 5-6 by hydrochloric acid (1M), then extracted with
ethyl acetate (20 mL.times.3). The combined organic layers were
washed with brine (20 mL.times.3), dried over anhydrous sodium
sulfate, filtered and concentrated under reduced pressure to give a
compound
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[7-[4-(3-hydroxypropoxy)b-
utylamino]-1-oxo-isoindolin-2-yl]acetic acid (0.35 g, 0.65 mmol,
70% yield, 98% purity) as a yellow oil. LCMS: MS (ESI) m/z: 531.3
[M+1].sup.+ .sup.1H NMR: (400 MHz, DMSO-d.sub.6) .delta.: 7.28-7.31
(m, 1H), 7.07-7.24 (m, 3H), 6.59-6.63 (m, 3H), 6.07-6.09 (m, 1H),
5.38-5.67 (m, 1H), 4.50-4.57 (m, 1H), 4.38 (t, J=5.2 Hz, 1H),
3.73-3.95 (m, 2H), 3.40-3.47 (m, 6H), 3.17-3.25 (m, 2H), 1.57-1.81
(m, 12H). Chemical Formula: C.sub.28H.sub.35FN.sub.2O.sub.7,
Molecular Weight: 530.59
13. Step--Synthesis of
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[7-[4-(3-hydroxypropoxy)b-
utylamino]-1-oxo-isoindolin-2-yl]-Nthiazol-2-yl-acetamide
##STR00797##
[1846] To a solution of
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[7-[4-(3-hydroxypropoxy)b-
utylamino]-1-oxo-isoindolin-2-yl]acetic acid (0.35 g, 0.65 mmol, 1
eq) in N,N-dimethylformamide (5 mL) was added
diisopropylethyllamine (167 mg, 1.29 mmol, 2 eq) and
0-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (491 mg, 1.29 mmol, 2 eq), then thiazol-2-amine
(97 mg, 0.97 mmol, 1.5 eq) was added, the mixture was stirred at
25.degree. C. for 1 hour. LCMS showed the starting material was
consumed completely and one main peak with desired m/z was
detected. The reaction mixture was concentrated under reduced
pressure. The residue was diluted with water (20 mL), then
extracted with ethyl acetate (20 mL.times.3). The combined organic
layers were washed with brine (20 mL.times.3), dried over anhydrous
sodium sulfate, filtered and concentrated under reduced pressure to
give a residue. The residue was purified by preparative Thin layer
chromatography (Petroleum ether/Ethyl acetate=1/1, Rf=0.24) to give
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[7-[4-(3-hydroxypropoxy)b-
utylamino]-1-oxo-isoindolin-2-yl]-N-thiazol-2-yl-acetamide (0.3 g,
0.49 mmol, 75% yield) as a yellow oil. LCMS: MS (ESI) m/z: 613.1
[M+1].sup.+ .sup.1H NMR: (400 MHz, CDCl.sub.3) .delta.: 7.42-7.46
(m, 1H), 7.30-7.35 (m, 1H), 7.12-7.26 (m, 2H), 6.93-7.09 (m, 2H),
6.42-6.71 (m, 4H), 5.17-5.52 (m, 1H), 4.59-4.82 (m, 1H), 4.07-4.15
(m, 1H), 3.77-3.79 (m, 2H), 3.61-3.64 (m, 2H), 3.39-3.54 (m, 4H),
3.25-3.29 (m, 2H), 1.74-1.89 (m, 12H). Chemical Formula:
C.sub.31H.sub.37FN.sub.4O.sub.6S, Molecular Weight: 612.71
14. Step--Synthesis of
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[1-oxo-7-[4-(3-oxopropoxy-
)butylamino]isoindolin-2-yl]-N-thiazol-2-yl-acetamide
##STR00798##
[1848] To a solution of
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[7-[4-(3-hydroxypropoxy)b-
utylamino]-1-oxo-isoindolin-2-yl]-N-thiazol-2-yl-acetamide (300 mg,
0.49 mmol, 1 eq) in dichloromethane (50 mL) was added
1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (311 mg,
0.73 mmol, 1.5 eq). The mixture was stirred at 25.degree. C. for
0.5 hour. High Performance Liquid Chromatography showed the
starting material was consumed completely. The reaction mixture was
filtered and concentrated under reduced pressure to give a residue.
The residue was purified by preparative Thin layer chromatography
(Petroleum ether/Ethyl acetate=1/1, Rf=0.14) to give
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[1-oxo-7-[4-(3-oxopropoxy-
)butylamino]isoindolin-2-yl]-N-thiazol-2-yl-acetamide (160 mg, 0.26
mmol, 53% yield) as a yellow solid. .sup.1H NMR: (400 MHz,
CDCl.sub.3) .delta.: 9.81 (s, 1H), 7.43-7.47 (m, 1H), 7.30-7.35 (m,
1H), 7.13-7.25 (m, 2H), 7.02-7.10 (m, 1H), 6.97-7.00 (m, 1H),
6.53-6.63 (m, 4H), 5.15-5.52 (m, 1H), 4.58-4.84 (m, 1H), 4.04-4.16
(m, 1H), 3.78 (t, J=6.4 Hz, 2H), 3.42-3.50 (m, 4H), 3.24-3.28 (m,
2H), 2.68 (t, J=6.4 Hz, 2H), 1.66-1.85 (m, 10H). Chemical Formula:
C.sub.31H.sub.35FN.sub.4O.sub.6S, Molecular Weight: 610.70
15. Step--Synthesis of tert-butyl
4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]-
piperidine-1-carboxylate
##STR00799##
[1850] To a solution of
2-(2,6-dioxo-3-piperidyl)-5-piperazin-1-yl-isoindoline-1,3-dione (1
g, 2.64 mmol, 1 eq, hydrochloride) in dichloromethane (5 mL) and
methanol (5 mL) was added sodium acetate (866 mg, 10.56 mmol, 4
eq), the mixture was stirred at 25.degree. C. for 1 hour, then
tert-butyl 4-oxopiperidine-1-carboxylate (526 mg, 2.64 mmol, 1 eq)
was added, the mixture was stirred at 25.degree. C. for 4 hours,
then sodium cyanoborohydride (331 mg, 5.28 mmol, 2 eq) was added,
the mixture was stirred at 25.degree. C. for another 7 hours. The
desired MS was observed by LCMS. The mixture was concentrated under
reduced pressure. The residue was purified by preparative High
Performance Liquid Chromatography (column: Phenomenex Synergi
Max-RP 250*50 mm*10 um; mobile phase: [water (0.225% FA)-ACN]; B %:
20ACN %-50ACN %, 30 min, 87% min) to afford tert-butyl
4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]-
piperidine-1-carboxylate (0.68 g, 1.29 mmol, 49.01% yield) as a
yellow solid. LCMS: MS (ESI) m/z: 526.3 [M+1].sup.+. .sup.1H NMR:
(400 MHz, MeOD) .delta.: 8.24 (s, 1H), 7.71 (d, J=8.4 Hz, 1H), 7.40
(d, J=2.4 Hz, 1H), 7.27 (dd, J=8.4, 2.4 Hz, 1H), 5.05-5.10 (m, 1H),
4.15-4.20 (m, 2H), 3.50-3.61 (m, 4H), 2.95-3.05 (m, 4H), 2.63-2.88
(m, 6H), 2.06-2.17 (m, 1H), 1.98-2.05 (m, 2H), 1.47-1.53 (m, 2H),
1.46 (s, 9H). Chemical Formula: C.sub.27H.sub.35N.sub.5O.sub.6,
Molecular Weight: 525.60
16. Step--Synthesis of 2-(2,6-dioxo-3-piperidyl)-5-[4-(4-piperidyl)
piperazin-1-yl]isoindoline-1,3-dione
##STR00800##
[1852] A solution of tert-butyl
4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]-
piperidine-1-carboxylate (0.54 g, 1.03 mmol, 1 eq) in hydrochloric
acid/1,4-dioxane (4 M, 10 mL) was stirred at 25.degree. C. for 1
hour. The desired MS was observed by LCMS. The mixture was
concentrated under reduced pressure to afford
2-(2,6-dioxo-3-piperidyl)-5-[4-(4-piperidyl)piperazin-1-yl]isoindoline-1,-
3-dione (0.48 g, 0.96 mmol, 94% yield, 93% purity, hydrochloride)
as a yellow solid. LCMS: MS (ESI) m/z: 426.2 [M+1].sup.+. Chemical
Formula: C.sub.22H.sub.27N.sub.5O.sub.4, Molecular Weight:
425.48
17. Step--Synthesis of
2-[7-[4-[3-[4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]pip-
erazin-1-yl]-1-piperidyl]propoxy]butylamino]-1-oxo-isoindolin-2-yl]-2-(5-f-
luoro-2-tetrahydropyran-2-yloxy-phenyl)-N-thiazol-2-yl-acetamide
##STR00801##
[1854] To a solution of
2-(2,6-dioxo-3-piperidyl)-5-[4-(4-piperidyl)piperazin-1-yl]isoindoline-1,-
3-dione (83 mg, 0.18 mmol, 1 eq, hydrochloride) in dichloromethane
(0.5 mL) and methanol (0.5 mL) was added sodium acetate (59 mg,
0.72 mmol, 4 eq), the mixture was stirred at 25.degree. C. for 0.5
hour, then
2-(5-fluoro-2-tetrahydropyran-2-yloxy-phenyl)-2-[1-oxo-7-[4-(3-oxopropoxy-
)butylamino]isoindolin-2-yl]-N-thiazol-2-yl-acetamide (110 mg, 0.18
mmol, 1 eq) was added, the mixture was stirred for 0.5 hour, then
sodium cyanoborohydride (22 mg, 0.36 mmol, 2 eq) was added, the
mixture was stirred at 25.degree. C. for 0.5 hour. LCMS showed the
starting material was consumed completely and one main peak with
desired m/z was detected. The reaction mixture was concentrated
under reduced pressure. The residue was diluted with water (20 mL),
then extracted with ethyl acetate (10 mL.times.3). The combined
organic layers were washed with brine (10 mL.times.3), dried over
anhydrous sodium sulfate, filtered and concentrated under reduced
pressure to give a residue. The residue was purified by preparative
Thin Layer Chromatography (dichloromethane/methanol=10/1, Rf=0.24)
to give
2-[7-[4-[3-[4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]pip-
erazin-1-yl]-1-piperidyl]propoxy]butylamino]-1-oxo-isoindolin-2-yl]-2-(5-f-
luoro-2-tetrahydropyran-2-yloxy-phenyl)-N-thiazol-2-yl-acetamide
(40 mg, 0.04 mmol, 21% yield) as a yellow solid. LCMS: MS (ESI)
m/z: 1020.2 [M+1].sub.+. Chemical Formula:
C.sub.53H.sub.62FN.sub.9O.sub.9S, Molecular Weight: 1020.18
18. Step--Synthesis of
2-[7-[4-[3-[4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]pip-
erazin-1-yl]-1-piperidyl]propoxy]butylamino]-1-oxo-isoindolin-2-yl]-2-(5-f-
luoro-2-hydroxy-phenyl)-N-thiazol-2-yl-acetamide
##STR00802##
[1856] To a solution of
2-[7-[4-[3-[4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]pip-
erazin-1-yl]-1-piperidyl]propoxy]butylamino]-1-oxo-isoindolin-2-yl]-2-(5-f-
luoro-2-tetrahydropyran-2-yloxy-phenyl)-N-thiazol-2-yl-acetamide
(0.04 g, 0.04 mmol, 1 eq) in ethyl acetate (0.5 mL) was added
Hydrochloric acid/Ethyl acetate (4 M, 0.5 mL). The mixture was
stirred at 25.degree. C. for 0.5 hour. LCMS showed one main peak
with desired m/z was detected. The reaction mixture was
concentrated under reduced pressure. The residue was purified by
preparative High Performance Liquid Chromatography (Formic acid as
additive) to afford
2-[7-[4-[3-[4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]pip-
erazin-1-yl]-1-piperidyl]propoxy]butylamino]-1-oxo-isoindolin-2-yl]-2-(5-f-
luoro-2-hydroxy-phenyl)-N-thiazol-2-yl-acetamide (19.2 mg, 0.02
mmol, 49% yield, 98% purity, formate) as a yellow solid. LCMS: MS
(ESI) m/z: 936.2 [M+1].sup.+ .sup.1H NMR: (400 MHz, CDCl.sub.3)
.delta.: 8.64 (s, 1H), 7.70 (d, J=8.8 Hz, 1H), 7.42 (d, J=3.6 Hz,
1H), 7.28-7.33 (m, 1H), 7.10-7.15 (m, 1H), 7.02-7.05 (m, 1H), 6.98
(d, J=3.6 Hz, 1H), 6.90-6.94 (m, 2H), 6.72 (s, 1H), 6.53-6.60 (m,
2H), 6.48 (d, J=8.4 Hz, 1H), 4.86-4.98 (m, 2H), 4.13 (d, J=17.2 Hz,
1H), 3.45-3.55 (m, 5H), 3.35-3.42 (m, 4H), 3.22-3.28 (m, 2H),
3.10-3.18 (m, 2H), 2.67-2.94 (m, 6H), 2.62-2.66 (m, 4H), 2.42-2.47
(m, 2H), 1.95-2.11 (m, 8H), 1.68-1.75 (m, 2H). Chemical Formula:
C.sub.48H.sub.54FN.sub.9O.sub.8S, Molecular Weight: 936.06
Synthesis of Example 320
(2S,4R)-1-((S)-18-(tert-butyl)-1-(5-(4-((3-chloro-4-((3-fluorobenzyl)oxy)p-
henyl)amino)quinazolin-6-yl)furan-2-yl)-16-oxo-2,5,8,11,14-pentaoxa-17-aza-
nonadecan-19-oyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine--
2-carboxamide
##STR00803##
[1858] Synthetic Scheme:
##STR00804##
1. Step--Synthesis of
2-[2-[2-[2-[2-[[5-[4-[3-Chloro-4-[(3-fluorophenyl)methoxy]anilino]quinazo-
lin-6-yl]-2-furyl]methoxy]ethoxy]ethoxy]ethoxy]ethoxy]acetic
acid
##STR00805##
[1860] To a solution of
[5-[4-[3-chloro-4-[(3-fluorophenyl)methoxy]anilino]quinazolin-6-yl]-2-fur-
yl]methanol (1) (47.59 mg, 0.1 mmol) in N,N-Dimethylformamide (1.5
ml) was added NaH (60%, 13 mg, 0.32 mmol) at room temperature. The
reaction mixture was stirred for 30 min. at the same temperature.
Then tert-butyl
2-[2-[2-[2-[2-(p-tolylsulfonyloxy)-ethoxy]ethoxy]ethoxy]ethoxy]acetate
(69.38 mg, 0.15 mmol) was added and the reaction mixture was
stirred for 20 min. at room temperature and then stirred for 12 h
(overnight) at 70.degree. C. external temperature (oil bath).
Solvent was removed under high vacuum and crude product was
purified by PTLC (DCM:MeOH:NH.sub.4OH, 90:9:1), to give 15 mg of
product
2-[2-[2-[2-[2-[[5-[4-[3-Chloro-4-[(3-fluorophenyl)methoxy]anilino]quinazo-
lin-6-yl]-2-furyl]methoxy]ethoxy]ethoxy]ethoxy]ethoxy]acetic acid.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.71 (s, 1H), 8.94 (s, 1H),
8.54 (s, 1H), 8.15 (dd, J=8.7, 1.6 Hz, 1H), 8.08 (d, J=2.4 Hz, 1H),
7.84-7.74 (m, 2H), 7.47 (td, J=8.0, 6.0 Hz, 1H), 7.37-7.28 (m, 3H),
7.25 (d, J=9.1 Hz, 1H), 7.18 (td, J=8.7, 2.2 Hz, 1H), 7.11 (d,
J=3.3 Hz, 1H), 6.64 (d, J=3.3 Hz, 1H), 5.25 (s, 2H), 4.55 (s, 2H),
3.76 (s, 2H), 3.70-3.39 (m, 16H). LC-MS (ESI); m/z [M+H]+: Calcd.
for C.sub.36H.sub.38ClFN.sub.3O.sub.9, 710.2280. Found
710.2403.
2. Step--Synthesis of
(2S,4R)-1-((S)-18-(tert-butyl)-1-(5-(4-((3-chloro-4-((3-fluorobenzyl)oxy)-
phenyl)amino)quinazolin-6-yl)furan-2-yl)-16-oxo-2,5,8,11,14-pentaoxa-17-az-
anonadecan-19-oyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-
-2-carboxamide
##STR00806##
[1862] To a solution of
2-[2-[2-[2-[2-[[5-[4-[3-chloro-4-[(3-fluorophenyl)methoxy]anilino]quinazo-
lin-6-yl]-2-furyl]methoxy]ethoxy]ethoxy]ethoxy]ethoxy]acetic acid
(3) (10 mg, 0.01 mmol) and
(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthia-
zol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (5) (7 mg,
0.015 mmol) in N,N-Dimethylformamide (2 ml) was added
N,N-Diisopropylethylamine (0.1 ml, 0.6 mmol) and
O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (8 mg, 0.021 mmol) at room temperature. The
reaction mixture was stirred for 12 h (overnight) at the same
temperature. TLC (DCM:MeOH:NH.sub.4OH, 90:9:1) shows no starting
materials. Reaction mixture was diluted with AcOEt (20 mL), washed
with water (4.times.15 mL), dried (Na.sub.2SO.sub.4) and evaporated
under vacuum. Crude product was purified by PTLC
(DCM:MeOH:NH.sub.4OH, 90:9:1), to give 7 mg of product
(2S,4R)-1-((S)-18-(tert-butyl)-1-(5-(4-((3-chloro-4-((3-fluoroben-
zyl)oxy)phenyl)amino)quinazolin-6-yl)furan-2-yl)-16-oxo-2,5,8,11,14-pentao-
xa-17-azanonadecan-19-oyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyr-
rolidine-2-carboxamide (44% yield). .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 9.98 (s, 1H), 8.97 (s, 1H), 8.76 (s, 1H), 8.67-8.57 (m,
1H), 8.55 (s, 1H), 8.17 (d, J=8.5 Hz, 1H), 8.00 (s, 1H), 7.80 (d,
J=8.7 Hz, 1H), 7.74 (d, J=10.3 Hz, 1H), 7.64-7.24 (m, 9H), 7.19 (t,
J=8.2 Hz, 1H), 7.09 (d, J=2.8 Hz, 1H), 6.65 (d, J=2.7 Hz, 1H), 5.26
(s, 2H), 5.15 (d, 1H), 4.61-4.54 (m, 1H), 4.54 (s, 2H), 4.49-4.15
(m, 6H), 3.94 (s, 2H), 3.73-3.40 (m, 17H), 2.43 (s, 3H), 2.11-2.01
(m, 1H), 1.95-1.84 (m, 1H), 0.93 (s, 9H). LC-MS (ESI); m/z
[M+H].sup.+: Calcd. for C.sub.58H.sub.66ClFN.sub.7O.sub.11S
1122.4213. Found 1122.0.
Synthesis of Example 349
1-(5-(4-((3-chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)quinazolin-6-yl)fur-
an-2-yl)-N-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)e-
thyl)-2,5,8,11,14-pentaoxahexadecan-16-amide
##STR00807##
[1864] Synthetic Scheme:
##STR00808##
1. Step--Synthesis of
1-(5-(4-((3-chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)quinazolin-6-yl)fu-
ran-2-yl)-N-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-
ethyl)-2,5,8,11,14-pentaoxahexadecan-16-amide
[1865] To a solution of
2-[2-[2-[2-[2-[[5-[4-[3-chloro-4-[(3-fluorophenyl)methoxy]anilino]quinazo-
lin-6-yl]-2-furyl]methoxy]ethoxy]ethoxy]ethoxy]ethoxy]acetic acid
(7 mg, 0.01 mmol) and
4-(2-aminoethoxy)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione
2,2,2-trifluoroacetate salt (6.38 mg, 0.01 mmol) in
N,N-Dimethylformamide (2 ml) was added N,N-Diisopropylethylamine
(0.1 ml, 0.6 mmol) and
O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (5.62 mg, 0.01 mmol) at room temperature. The
reaction mixture was stirred for 12 h (overnight) at the same
temperature. TLC (DCM:MeOH:NH.sub.4OH, 90:9:1) shows no starting
materials. Reaction mixture was diluted with AcOEt (20 mL), washed
with water (4.times.15 mL), dried (Na.sub.2SO.sub.4) and evaporated
under vacuum. Crude product was purified by PTLC
(DCM:MeOH:NH.sub.4OH, 90:9:1), to give 7.2 mg of product
1-(5-(4-((3-chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)quinazolin-6-yl)fu-
ran-2-yl)-N-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-
ethyl)-2,5,8,11,14-pentaoxahexadecan-16-amide (72% yield). .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 10.27 (s, 1H), 9.14 (s, 1H), 7.93
(s, 1H), 7.72 (s, 1H), 7.34 (dd, J=8.8, 1.7 Hz, 1H), 7.18 (d, J=2.6
Hz, 1H), 7.08-6.87 (m, 4H), 6.76-6.56 (m, 3H), 6.56-6.42 (m, 3H),
6.36 (td, J=8.8, 2.3 Hz, 1H), 6.26 (d, J=3.3 Hz, 1H), 5.82 (d,
J=3.3 Hz, 1H), 4.44 (s, 2H), 4.25 (dd, J=12.7, 5.4 Hz, 1H), 3.71
(s, 2H), 3.43 (t, J=6.0 Hz, 2H), 2.87-2.58 (m, 16H), 2.12-1.99 (m,
1H), 1.80-1.60 (m, 2H), 1.23-1.13 (m, 1H). .sup.13C NMR (151 MHz,
DMSO-d6) .delta. 172.76, 169.89, 169.70, 166.76, 165.19, 163.00,
161.39, 157.63, 155.56, 154.37, 152.52, 152.30, 149.79, 149.03,
139.68, 139.63, 136.97, 133.23, 133.03, 130.60, 130.54, 128.82,
128.52, 128.05, 124.42, 123.35, 123.33, 122.62, 121.00, 119.98,
116.70, 116.48, 115.52, 115.32, 114.77, 114.63, 114.25, 114.11,
113.97, 112.24, 107.89, 70.21, 69.86, 69.77, 69.74, 69.72, 69.50,
69.39, 69.38, 68.89, 67.09, 64.17, 48.74, 37.35, 30.94, 22.00.
LC-MS (ESI); m/z [M+H].sup.+: Calcd. for
C.sub.51H.sub.51ClFN.sub.6O.sub.3, 1009.3186. Found 1009.3224.
Synthesis of examples 350 and 351
##STR00809##
[1866] tert-Butyl 3-(3-(3-chloropropoxy)propoxy)propanoate (1)
[1867] 3-(3-chloropropoxy)propan-1-ol (66 mg, 0.43 mmol) in
acetonitrile (3 mL) was added tert-butyl prop-2-enoate (0.31 ml,
2.16 mmol) followed by Triton B (54 mg, 0.1 mmol, 40% by weight in
water). The mixture was stirred at room temperature for 72 hour.
The mixture was concentrated under vacuum and crude product was
purified by column chromatography (SiO.sub.2, gradient Hex:EtOAc,
95:5 to 9:1) to give 115 mg of product (1) as an oil (94% yield).
.sup.1H NMR (500 MHz, Chloroform-d) .delta. 3.70-3.59 (m, 4H),
3.59-3.42 (m, 6H), 2.47 (t, J=6.5 Hz, 2H), 2.04-1.96 (m, 2H), 1.82
(p, J=6.3 Hz, 2H), 1.45 (s, 9H). .sup.13C NMR (151 MHz,
Chloroform-d) .delta. 171.13, 80.63, 68.02, 67.97, 67.27, 66.64,
42.17, 36.50, 32.88, 30.09, 28.25. LC-MS (ESI); m/z [M+Na].sup.+:
Calcd. for C.sub.13H.sub.25ClO.sub.4Na, 303.1339. Found
303.1381.
tert-Butyl 3-(3-(3-iodopropoxy)propoxy)propanoate (2)
[1868] To a solution of tert-butyl
3-[3-(3-chloropropoxy)propoxy]propanoate (161 mg, 0.57 mmol) in
Acetone (5 ml) was added NaI (429 mg, 2.87 mmol). The reaction
mixture was stirred at reflux temperature for 24 h, then the
solvent was removed under vacuum and crude product was dissolved in
EtOAc (15 mL), washed with water (10 mL), and with an aqueous
solution of Na.sub.2SO.sub.3 (10%, 10 mL). Organic layer was
separated, washed with water (10 mL), dried (Na.sub.2SO.sub.4) and
evaporated under vacuum. Crude product was pure by NMR (>98%
purity, 186 mg, 87% yield), product (2) was used in the next step
without any further purification. .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 3.66 (t, J=6.5 Hz, 2H), 3.57-3.40 (m, 6H),
3.27 (t, J=6.8 Hz, 2H), 2.48 (t, J=6.5 Hz, 2H), 2.08-1.99 (m, 2H),
1.82 (p, J=6.4 Hz, 2H), 1.45 (s, 9H). .sup.13C NMR (151 MHz,
Chloroform-d) .delta. 171.13, 80.64, 70.18, 68.01, 67.98, 66.65,
36.50, 33.57, 30.10, 28.26, 3.72. LC-MS (ESI): m/z [M+Na].sup.+
Calcd. for C.sub.13H.sub.25O.sub.4Na: 395.0695, Found:
395.0719.
##STR00810##
[1869] tert-Butyl
3-(3-(3-((4-(2-fluoro-4-(1-((4-fluorophenyl)carbamoyl)cyclopropane-1-carb-
oxamido)-phenoxy)-6-methoxyquinolin-7-yl)oxy)propoxy)propoxy)propanoate
(12).
[1870] To a mixture of
N1'-[3-fluoro-4-[(7-hydroxy-6-methoxy-4-quinolyl)oxy]phenyl]-N1-(4-fluoro-
phenyl)cyclopropane-1,1-dicarboxamide (11) (15 mg, 0.03 mmol) and
tert-butyl 3-[3-(3-iodopropoxy)propoxy]propanoate (2) (16.57 mg,
0.04 mmol) in N,N-Dimethylformamide (1 mL) was added
Cs.sub.2CO.sub.3 (29.01 mg, 0.09 mmol). After stirring at room
temperature for 12 hrs (overnight), the reaction mixture was
diluted with AcOEt (20 mL) and washed with water (5.times.10 mL),
organic phase was evaporated under vacuum. Crude product was
purified by PTLC (DCM:MeOH:NH.sub.4OH, 92:7:1) to give 15 mg of
product (12) (67% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.
10.39 (s, 1H), 10.01 (s, 1H), 8.46 (d, J=5.2 Hz, 1H), 7.90 (d,
J=13.2 Hz, 1H), 7.71-7.58 (m, 2H), 7.51 (d, J=7.4 Hz, 2H),
7.46-7.35 (m, 2H), 7.15 (t, J=8.9 Hz, 2H), 6.41 (d, J=5.1 Hz, 1H),
4.21 (t, J=6.2 Hz, 2H), 3.95 (s, 3H), 3.60-3.37 (m, 8H), 2.37 (d,
J=12.2 Hz, 2H), 2.04 (p, J=6.4 Hz, 2H), 1.71 (p, J=6.4 Hz, 2H),
1.47 (s, 4H), 1.37 (s, 9H). .sup.13C NMR (151 MHz, DMSO-d6) .delta.
170.45, 168.27, 167.87, 159.29, 159.07, 157.48, 154.07, 152.44,
151.89, 149.56, 148.82, 146.37, 138.05, 137.98, 135.70, 135.61,
135.20, 135.19, 123.82, 122.46, 122.41, 116.90, 115.11, 115.09,
114.96, 114.47, 109.04, 108.88, 108.50, 101.95, 99.01, 79.64,
67.07, 66.55, 65.92, 65.45, 55.79, 35.87, 31.93, 29.53, 28.90,
27.76, 27.73, 15.31. LC-MS (ESI): m/z [M+H].sup.+ Calcd. for
C.sub.40H.sub.46F.sub.2N.sub.3O.sub.9, 750.3202. Found
750.3509.
##STR00811##
N-(3-Fluoro-4-((7-(3-(3-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthia-
zol-5-yl)benzyl)carbamoyl)-pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)a-
mino)-3-oxopropoxy)propoxy)propoxy)-6-methoxyquinolin-4-yl)oxy)phenyl)-N-(-
4-fluorophenyl)cyclopropane-1,1-dicarboxamide (SJF-8240, PROTAC
7)
[1871] A solution of tert-butyl
3-[3-[3-[[4-[2-fluoro-4-[[1-[(4-fluorophenyl)carbamoyl]cyclopropanecarbon-
yl]amino]phenoxy]-6-methoxy-7-quinolyl]oxy]propoxy]propoxy]propanoate
(12) (15 mg, 0.02 mmol) in a mixture of TFA (1 ml, 13.46 mmol) and
Dichloromethane (3 ml) was stirred for 2 h. Then the solvent was
removed under vacuum and crude product was dried under high vacuum
for 2 h. Crude product was used in the next step without any
further purification (13.8 mg, quantitative yield). LC-MS (ESI):
m/z [M+H]+ Calcd. for C.sub.36H.sub.38F.sub.2N.sub.3O.sub.9,
694.2576. Found 694.2324. To a solution of crude product from above
(13.8 mg, 0.02 mmol) and
(2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[4-(4-methylt-
hiazol-5-yl)phenyl]methyl]-pyrrolidine-2-carboxamide; hydrochloride
(8) (11.15 mg, 0.02 mmol) in N,N-Dimethylformamide (2 ml) was added
DIPEA (0.17 ml, 0.99 mmol) and HATU (11.35 mg, 0.03 mmol) at room
temperature. The reaction mixture was stirred for 12 h (overnight)
at the same temperature. Reaction mixture was diluted with ACOEt
(20 mL), washed with water (4.times.15 mL), dried
(Na.sub.2SO.sub.4) and evaporated under vacuum. Crude product was
purified by PTLC (DCM:MeOH:NH.sub.4OH, 90:9:1), to give 18 mg of
product (82% yield). .sup.1H NMR (500 MHz, DMSO-d6) .delta. 10.38
(s, 1H), 10.00 (s, 1H), 8.97 (s, 1H), 8.56 (t, J=6.1 Hz, 1H), 8.46
(d, J=5.2 Hz, 1H), 7.96-7.85 (m, 2H), 7.69-7.59 (m, 2H), 7.51 (d,
J=8.8 Hz, 2H), 7.45-7.33 (m, 5H), 7.15 (t, J=8.9 Hz, 2H), 6.41 (d,
J=5.1 Hz, 1H), 5.12 (d, J=3.3 Hz, 1H), 4.55 (d, J=9.4 Hz, 1H), 4.43
(ddd, J=10.9, 6.7, 3.3 Hz, 2H), 4.27-4.16 (m, 3H), 3.94 (s, 3H),
3.76-3.33 (m, 10H), 2.58-2.51 (m, 1H), 2.43 (s, 3H), 2.35-2.25 (m,
1H), 2.03 (p, J=5.7 Hz, 3H), 1.95-1.83 (m, 1H), 1.72 (p, J=6.4 Hz,
2H), 1.48 (d, J=3.9 Hz, 4H), 0.92 (s, 9H). .sup.13C NMR (126 MHz,
DMSO-d6) .delta. 171.89, 169.97, 169.51, 168.26, 167.88, 159.31,
159.22, 157.31, 154.21, 152.26, 151.90, 151.39, 149.56, 148.75,
147.69, 146.29, 139.47, 138.01, 137.94, 135.70, 135.60, 135.17,
135.15, 131.13, 129.61, 128.81, 128.61, 127.40, 123.77, 122.46,
122.40, 116.90, 115.09, 114.92, 114.47, 109.53, 109.05, 108.87,
108.45, 101.94, 99.03, 68.85, 67.16, 67.09, 66.62, 66.54, 65.47,
58.69, 56.35, 56.24, 55.77, 41.64, 37.92, 35.69, 35.36, 31.87,
29.60, 28.89, 26.28, 15.91, 15.31. LC-MS (ESI): m/z [M+H].sup.+
Calcd. for C.sub.58H.sub.66F.sub.2N.sub.7O.sub.11S, 1106.4509.
Found 1106.4510.
N-(3-Fluoro-4-((7-(3-(3-(3-(((S)-1-((2S,4S)-4-hydroxy-2-((4-(4-methylthiaz-
ol-5-yl)benzyl)carbamoyl)-pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)am-
ino)-3-oxopropoxy)propoxy)propoxy)-6-methoxyquinolin-4-yl)oxy)phenyl)-N-(4-
-fluorophenyl)cyclopropane-1,1-dicarboxamide (SJF-8240-epimer,
PROTAC 8)
[1872] It was prepared from
(2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[4-(4-methylt-
hiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide; hydrochloride
(10) (10.42 mg, 0.022 mmol) and following the same procedure than
above. Crude product was purified by PTLC (DCM:MEOH:NH.sub.4OH,
90:9:1), to give 9.7 mg of the expected product (47% yield).
.sup.1H NMR (500 MHz, DMSO-d6) .delta. 10.38 (s, 1H), 10.00 (s,
1H), 8.97 (s, 1H), 8.63 (t, J=6.0 Hz, 1H), 8.46 (d, J=5.2 Hz, 1H),
7.90 (d, J=9.7 Hz, 2H), 7.71-7.57 (m, 2H), 7.51 (d, J=8.7 Hz, 2H),
7.44-7.28 (m, 5H), 7.15 (t, J=8.9 Hz, 2H), 6.41 (d, J=5.1 Hz, 1H),
5.43 (d, J=7.2 Hz, 1H), 4.56-4.38 (m, 2H), 4.36 (dd, J=8.6, 6.1 Hz,
1H), 4.32-4.13 (m, 4H), 3.94 (s, 3H), 3.97-3.82 (m, 1H), 3.64-3.46
(m, 4H), 3.48-3.35 (m, 4H), 2.57-2.45 (m, 2H), 2.43 (s, 3H),
2.36-2.26 (m, 2H), 2.03 (p, J=6.3 Hz, 2H), 1.73 (dp, J=13.0, 6.2
Hz, 3H), 1.55-1.38 (m, 4H), 0.93 (s, 9H). .sup.13C NMR (126 MHz,
DMSO-d6) .delta. 171.89, 169.97, 169.51, 168.26, 167.88, 159.31,
159.22, 157.31, 154.21, 152.26, 151.90, 151.39, 149.56, 148.75,
147.69, 146.29, 139.47, 138.01, 137.94, 135.70, 135.60, 135.17,
135.15, 131.13, 129.61, 128.81, 128.61, 127.40, 123.77, 122.46,
122.40, 116.90, 115.09, 114.92, 114.47, 109.53, 109.05, 108.87,
108.45, 101.94, 99.03, 68.85, 67.16, 67.09, 66.62, 66.54, 65.47,
58.69, 56.35, 56.24, 55.77, 41.64, 37.92, 35.69, 35.36, 31.87,
29.60, 28.89, 26.28, 15.91, 15.31. LC-MS (ESI): m/z [M+H].sup.+
Calcd. for C.sub.58H.sub.66F.sub.2N.sub.7O.sub.11S, 1106.4509.
Found 1106.5096.
[1873] Protein Level Control
[1874] This description also provides methods for the control of
protein levels with a cell. This is based on the use of compounds
as described herein, which are known to interact with a specific
target protein such that degradation of a target protein in vivo
will result in the control of the amount of protein in a biological
system, preferably to a particular therapeutic benefit.
[1875] The following examples are used to assist in describing the
present invention, but should not be seen as limiting the present
invention in any way.
EXAMPLES
[1876] With reference to the appended figures and accompanying
descriptions.
[1877] FIGS. 1A and 1B provides an illustration of general
principle for PROTAC function. FIG. 1A provides exemplary PROTACs
comprise a protein targeting moiety (PTM; darkly shaded rectangle),
a ubiquitin ligase binding moiety (ULM; lightly shaded triangle),
and optionally a linker moiety (L; black line) coupling or
tethering the PTM to the ULM. FIG. 1B illustrates the functional
use of the PROTACs as described herein. Briefly, the ULM recognizes
and binds to a specific E3 ubiquitin ligase, and the PTM binds and
recruits a target protein bringing it into close proximity to the
E3 ubiquitin ligase. Typically, the E3 ubiquitin ligase is
complexed with an E2 ubiquitin-conjugating protein, and either
alone or via the E2 protein catalyzes attachment of ubiquitin (dark
circles) to a lysine on the target protein via an isopeptide bond.
The poly-ubiquitinated protein (far right) is then targeted for
degradation by the proteosomal machinery of the cell.
[1878] FIG. 2 provides a table of exemplary PROTAC compounds
(compounds 1-351) as described herein. The compounds are exemplary
and the description is expressly intended to encompass combinations
of the exemplified compounds respective PTM, Linker, and ULM
components. For example, the PTM of compound example 1, can be
combined with any of the linkers exemplified in examples 2-351
and/or any of the ULMs in examples 2-351, and so on.
[1879] Small molecule induced degradation of EGFR and mutants.
Epidermal Growth Factor Receptor (EGFR), also known as ErbB1/HER1,
is a proto-oncogene that has been implicated in a range of cancers
including glioblastoma multiforme, head and neck, and non-small
cell lung cancer. Overexpression and/or activating mutations of
EGFR are associated with a poor prognosis, therefore significant
effort has focused on targeting EGFR with both small molecule and
antibody-based therapies. Small molecule kinase inhibitors
competitively bind to the kinase domain, thereby preventing
signalling, while antibodies are capable of preferentially binding
the cognate ligand recognition site, thus preventing kinase
activation. Furthermore, degradation of EGFR by FDA-approved
antibodies has been implicated in their clinical success,
suggesting that degradation may be advantageous. With this
rationale in mind, small molecules capable of inducing EGFR
degradation were developed (i.e., EGFR-targeting PROTACs).
[1880] By conjugating an EGFR binding element, e.g., kinase
inhibitor, such as lapatinib (FIG. 3), to a ligand that binds to
the E3 ligase, VHL, exemplary molecules were synthesized capable of
penetrating the cell membrane and inducing EGFR degradation at low
nanomolar concentrations (FIGS. 4A-6C). Interestingly, inversion of
the hydroxyproline stereochemistry on the VHL-recruiting moiety of
the PROTAC can affect its ability to degrade EGFR (FIG. 4B). This
diastereomeric version (PROTAC Diastereomer 2) of the PROTAC
provides an excellent control compound with nearly identical
physicochemical properties (see FIGS. 6A-6C) but capable of only
inhibiting EGFR; an ideal tool with which to directly assess the
advantages of EGFR degradation over kinase inhibition without
variations in probe solubility, membrane permeability, or chemical
stability obscuring a head-to-head comparison (as would be the case
if comparisons were drawn directly with lapatinib itself).
[1881] Having demonstrated that recruitment of VHL to EGFR via a
lapatinib-based PROTAC is capable of efficiently inducing
degradation of a receptor tyrosine kinase (RTK), different
EGFR-binding elements were employed to degrade different clinically
relevant forms of EGFR. As shown in FIG. 4C, the lapatinib-based
PROTAC 1, is also capable of degrading an exon-20 insertion mutant
form of EGFR (ASV duplication). Switching to a warhead based on
mutant-EGFR selective gefitinib (PROTAC 3) enabled the degradation
of both exon-19 deletion EGFR as well as the mutant isoform
containing the L858R activating point mutation (FIG. 4D/4E), while
sparing the WT EGFR (see FIGS. 5A-5D). Finally employing the
second-generation inhibitor afatinib yielded PROTAC 4 capable of
degrading the gefitinib-resistant double mutant (L858R/T790M) EGFR
(FIG. 4F).
[1882] The choice of warhead can be crucial for successful target
degradation; here it is demonstrated that careful selection of the
recruiting element can also allow degradation of proteins in
different mutational states. For some of the aforementioned
PROTACs, a "hook effect" was observed on substrate degradation,
which has been previously reported and results from the formation
of unproductive dimers (rather than productive `trimers`) at higher
concentrations. The lack of this `hook effect` in other PROTACs
might arise from additive target: E3 ligase protein-protein
interactions that they induce.
[1883] Selective PROTAC-mediated degradation of HER2 and
implications for kinome re-wiring. Since lapatinib is also a potent
binder to other HER family RTKs, the potential for HER2 degradation
by lapatinib-based PROTACs was explored. Similar to EGFR, HER2
overexpression is an oncogenic driver of many forms of cancer
including ovarian, breast, and gastric cancers. Immunoblotting
analysis revealed that PROTAC 1, which utilizes a diethylene glycol
linker to tether lapatinib to the VHL recruiting element, could
concurrently degrade both EGFR and HER2 (FIG. 5A). However,
extension of the linker by an additional ethylene glycol unit to
create PROTAC 5 enabled the selective degradation of EGFR while
sparing HER2. This observation suggests that affinity for both
target protein and E3 ligase is not sufficient for the development
of a successful PROTAC, and that a more complex dynamic process may
be responsible.
[1884] The advantages of PROTAC-mediated degradation over kinase
inhibition become apparent when the effect on cell proliferation is
compared (FIG. 5B). SKBr3 cells, a HER2-driven breast cancer cell
line, are more responsive to PROTAC 1 than the cognate
diastereomeric control 2 that has equivalent cell permeability and
kinase inhibitory properties but is incapable of inducing HER2
degradation (FIG. 6B/6C). PROTAC 1 treatment induces a greater
response in terms of anti-proliferative efficacy than does
diastereomer 2, and at a greater potency (PROTAC 1 IC.sub.50=102
nM, diastereomer 2 IC.sub.50=171 nM).
[1885] Comparison of PROTAC 1 with diastereomer 2 also reveals
additional advantages of degradation over inhibition following an
extended treatment period. Inhibition of EGFR/HER2 in SKBr3 cells
has previously been shown to rapidly induce kinome "re-wiring",
whereby alternative, uninhibited kinases are recruited as
heterodimerization partners leading to the phosphorylation of the
same downstream effectors to restore oncogenic signalling via RTK
crosstalk.
[1886] To investigate this phenomenon, SKBr3 cells were treated
with equal, saturating concentrations (FIG. 6B/6C) of either PROTAC
1 or diastereomer 2, which provided a direct and head-to-head
comparison of degradation versus inhibition given their similar
physicochemical properties. Interestingly, degradation appears to
have a protective effect against kinome re-wiring, particularly in
the case of ERK1/2 phosphorylation and specific HER3 and AKT
phosphorylation sites. Cells treated with the inhibitor
(diastereomer) 2 led to transient suppression of activation of
ERK1/2, phosphorylation of Akt (Thr308), c-Met and HER3, the latter
being a major node for preventing apoptosis and promoting survival
(FIG. 5C). However, within 24 to 48 hours, this suppression was
reversed despite continued presence of the inhibitor.
[1887] Quite strikingly, treatment with an equivalent concentration
of PROTAC 1 itself yielded sustained suppression of downstream
signalling, suggesting that removal of the target RTKs discourages
kinome re-wiring and permits longer sustained growth suppression.
Inhibition transiently prevents downstream signalling but
degradation may also impact the scaffolding roles exhibited by
RTKs, particularly in instances of kinome re-wiring by receptor
cross-talk. For example, heterodimerization of EGFR with c-Met and
signalling via the c-Met kinase domain has been implicated in
resistance to some inhibitors/antibodies. Analysis of the c-Met
phosphorylation status after 48 hours of PROTAC 1 or diastereomer 2
treatment revealed a substantial increase in signalling via c-Met
kinase domain in diastereomer-treated cells compared to the PROTAC
treated cells, presumably by trans-activation (FIG. 5D). These
results demonstrate the advantages gained from RTK degradation
compared to kinase inhibition with regards to prevention of
downstream signalling and delayed onset of kinome re-wiring.
[1888] Since PROTACs proved successful in degrading both EGFR and
HER2, PROTACs were made to a different RTK family. C-Met is the
receptor for Hepatocyte Growth Factor (HGF), which is also known as
the "scatter factor" for its ability to promote tumor metastasis.
Upon binding of HGF, c-Met dimerizes and transphosphorylates on
tyrosine residues within its kinase domain (Y1234 and Y1235) as
well as on its unique c-terminal multifunctional docking domain
(Y1313, Y1349, Y1356 and Y1365). The docking domain contains
recognition sites for diverse cellular effectors such as Src, Gab1,
Crk, Grb2, SHC and PI-3 kinase, which potently activate downstream
mitogenic pathways. Inhibitors, such as foretinib, which
competitively displace ATP from the c-Met kinase domain block
HGF-stimulated activation of ERK and Akt, the primary downstream
effectors of c-Met signalling. Despite this, however, small
molecule c-Met inhibitors have performed disappointingly in
clinical trials suggesting the possibility of a kinase-independent
function driving oncogenesis and highlighting the potential
advantage of c-Met degradation over inhibition.
[1889] FIGS. 6A-6C demonstrate that a Gefitinib-based PROTAC
(PROTAC 3) spares WT EGFR (FIG. 6A). OVCAR8 Cells were treated for
24 hours with increasing doses of PROTAC 3 or with DMSO control
before immunoblotting. FIG. 6B/6C--Characterization of PROTAC 1
(6B) and diastereomer 2 (6C) in SKBr3 cells. Cells were treated for
24 hours in full serum with increasing doses of PROTAC 1 or with
diastereomer 2 before immunoblotting. The characterization of
foretinib-based PROTACs in GTL16 cells is shown in FIGS. 7A-7F.
GTL16 cells were treated with increasing concentrations of PROTAC 7
(7A) or diastereomer 8 (7B) in media containing full serum for 24
hours before immunoblotting analysis. FIG. 7C--Representative blot
of cells treated with 500 nM PROTAC 7 or 500 nM diastereomer 8 for
48 hr before immunoblotting analysis. FIG. 7D shows the
quantification of washout experiments. c-MET levels normalized to
tubulin after treatment with the indicated compounds at the
indicated time points. Average of 3 independent repeats and error
bars represent S.E.M. The structure of VHL-Ligand 9 used in
competition experiments is shown in FIG. 7E. FIG. 7F shows
co-treatment competition of PROTAC 7 with VHL-Ligand 9 in
MDA-MB-231 cells for 24 hours.
[1890] Employing the c-Met inhibitor foretinib as a recruiting
element, we developed a PROTAC, compound 7, capable of recruiting
VHL to, and thereby inducing degradation of c-Met in a dose- and
time-dependent fashion. MDA-MB-231 cells treated with increasing
concentrations of foretinib-based PROTAC 7 (see FIG. 3 and FIG. 8A)
or its cognate diastereomer 8 (see FIG. 3 and FIG. 8B), in the
presence or absence of an HGF pulse, demonstrated that treatment
with approximately 10-fold higher concentration of diastereomer is
required to completely inhibit agonist-driven AKT phosphorylation.
We observed a similar level of inhibition of Akt phosphorylation in
GTL16 cells, a c-Met-overexpressing cell line, when grown and
treated in full serum with PROTAC 7 and diastereomer 8. The reduced
potency differential between PROTAC 7 and diastereomer 8 inhibition
of Akt phosphorylation likely results from the steady-state versus
agonist-challenged activation of the signaling cascade between the
two cell lines (GTL16 and MDA-MB-231, respectively). The
foretinib-based PROTAC 7 is also greater than 2-fold more potent
than its corresponding diastereomer 8 at inhibiting the
proliferation of GTL16 cells (FIG. 8C), again highlighting the
advantages of developing probes capable of protein degradation,
especially in oncogene-addicted contexts. Additionally, clearance
of WT c-Met from the cell is relatively rapid, requiring only 6 hr
of treatment to significantly reduce protein levels in MDA-MB-231
cells (FIG. 8D), providing a temporal advantage over RNA-mediated
knockdown techniques as well as abrogating the requirement for
transfection reagents or exogenous selection pressure that may
interfere with other normal cellular activities.
[1891] Subsequent to demonstrating the catalytic nature of PROTACs,
the duration of their effect was studied. Cells were treated for 24
hr with DMSO control, foretinib-based PROTAC 7, or its
corresponding diastereomer 8 before being dissociated from the
culture dishes, rinsed with PBS to wash out extracellular treatment
compound, and replated into fresh medium and on new culture dishes
for additional 24- or 48-hr periods followed by lysis.
PROTAC-treated cells exhibited a prolonged reduction in c-Met
levels out to 48 hr post washout. Crucially, in cells treated with
PROTAC 7, c-Met levels could be rescued by treatment with 50-fold
excess free VHL ligand following the washout. The free VHL ligand
prevents E3 ligase recruitment to the RTK by PROTAC 7, indicating
that the sustained knockdown in PROTAC-treated cells is mediated
post-translationally by disrupting PROTAC 7:Met:VHL complexes
remaining in the cell (catalytic mode of action) rather than a
response at the translational level, as has been observed with
other small molecules. Moreover, when MDA-MB-231 cells are
co-treated with PROTAC 7 and increasing doses of free VHL ligand,
the ability of the PROTAC to degrade c-Met is hindered, further
demonstrating the necessity and specificity for VHL recruitment.
Additionally, foretinib-based PROTAC 7 is also capable of
preventing inhibitor-induced compensatory signaling in a way
similar to that of the aforementioned lapatinib-based PROTACs.
GTL16 cells display profound kinome rewiring after 48 hr, as
evidenced by ERK1/2 phosphorylation in diastereomer 8-treated
cells, but not in PROTAC 7-treated cells.
[1892] Having proven that RTKs could be degraded via the PROTAC
technology the mechanism of degradation was assessed. Initially,
real-time qPCR was performed over time to demonstrate that the
observed decrease in c-Met protein levels in response to PROTAC
treatment occurs at the post-transcriptional level. Additionally,
co-treatment with pharmacological agents that inhibit either the
proteasome (epoxomicin) or the ubiquitination cascade
(NEDD8-activating enzyme E1 inhibitor, MLN-4924) were able to
restore protein levels to untreated levels, demonstrating not only
that ubiquitination is crucial for c-Met degradation but also that
it progresses via the proteasome. Furthermore, RTKs are known to
rely on the heatshock protein 90 (HSP90) chaperone for proper
folding as well as being cycled between the plasma membrane and
early endosomes in a HSP90-dependent fashion.
[1893] As such the effect of the HSP90 inhibitor 17-AAG
(17-N-allylamino-17-demethoxygeldanamycin) on PROTAC-mediated
degradation was explored. Co-treatment with PROTAC 7 and 17-AAG has
an additive effect on c-Met protein degradation in the MDA-MB-231
cell line, suggesting that HSP90 may be intercepting c-Met in a
separate compartment from PROTAC 7, thereby enhancing the
degradation of total c-Met within the cell. Previous work has shown
c-Met to be a client protein of HSP90 and that geldanamycin and
17-AAG could promote its ubiquitination and proteasome-dependent
degradation.
[1894] It was then determined whether PROTAC-targeted RTKs were
removed directly from the cell surface or were intercepted at some
point in the secretory pathway to the membrane. Employing a
cell-surface biotinylation degradation assay, it was demonstrated
that PROTAC 7 induces the degradation of the c-Met mature form (145
kDa) from the cell surface, suggesting that VHL recruitment is
capable of, either directly or indirectly, inducing RTK
internalization (FIGS. 9A and 9B). We confirmed PROTAC 7-mediated
RTK internalization by confocal immunofluorescence microscopy:
untreated cells exhibit cell-surface c-Met immunofluorescence but
treatment with PROTAC 7 (FIG. 9C) or HGF shows internalization and
localization to a perinuclear compartment as previously described.
This perinuclear compartment also stains positive for the early
endosome antigen 1 (EEA1) and appears to be distinct from the Golgi
apparatus, which is c-Met positive in untreated cells (FIGS.
12A-12E). These large EEA1-positive vesicle-like structures have
previously been reported in MDA-MB-231 cells. This provides, to the
best of our knowledge, the first evidence of small-molecule-induced
internalization of an endogenous RTK and further suggests sorting
into endosomes prior to degradation via the proteasome.
Interestingly, preliminary small interfering RNA experiments
suggest that this process is clathrin independent (FIG. 12E).
[1895] Next, the advantages of degradation over inhibition was
assessed in the Hs746T gastric cancer cell line, which expresses an
exon 14 splice variant of c-Met. Exon 14 skipping results in the
expression of c-Met lacking he juxtamembrane domain recruitment
site (Y1003) for Cbl, the endogenous E3 ligase that promotes
HGF-dependent internalization and subsequent degradation of c-Met.
This clinically relevant mutation results in prolonged downstream
signaling, since the naturally occurring "off-switch" for
HGF-induced signaling is no longer present. The lack of this
regulatory domain also increases the intrinsic stability of c-Met
protein in the absence of any other degradation signal.
Cycloheximide chase experiments (FIG. 10A) revealed that WT c-Met
has a basal half-life of 4.4 hr, while the basal half-life of the
exon 14 mutant c-Met is >8 hr. HGF treatment results in rapid
degradation of WT c-Met (FIGS. 10A and 10C) but not exon 14-deleted
c-Met (FIGS. 10A, 10B, and 10D); this lack of
internalization/degradation results in sustained downstream
signaling (FIGS. 10C and 10D), enhancing the cell proliferation and
tumorigenesis of exon 14 mutant c-Met-expressing cells.
[1896] Interestingly, PROTAC 7 treatment can induce the degradation
of exon 14-deleted c-Met (FIGS. 10A and 10E) despite that fact that
it is not degraded by the major natural mechanism (i.e., HGF). The
degradation half-life of PROTAC 7-treated exon 14-deleted c-Met is
only marginally longer than that of PROTAC 7-treated WT c-Met (4.2
hr versus 2.5 hr, respectively), in contrast to the wide
differential in their respective HGF-induced, native degradation
rates (>8 hr versus 1.66 hr, respectively) (FIG. 10B). The fact
that PROTAC 7 is able to degrade exon 14-deleted c-Met suggests
that this process in Cbl independent as well as clathrin
independent.
[1897] This provided another instance whereby target degradation by
PROTAC might prove advantageous over inhibition alone, in that
inhibition can temporarily block signaling at the level of kinase
activity, but only degradation can provide a lasting "off-switch"
for the receptor itself as demonstrated in FIGS. 10F and 10G.
Pre-treatment of Hs746T cells, which express an exon 14 mutant
c-Met, with PROTAC 7 reduces HGF-induced activation of Akt. There
remains a brief elevation of phospho-Akt in these cells at 0.5 hr
following HGF treatment; more importantly, however, is that the
sustained signaling observed in DMSO-treated cells (up to 6 hr) is
not observed in the PROTAC 7-treated cells (FIG. 10G). This
restoration of a WT phenotype to a mutant protein via a PROTAC is
intriguing, and the approach could prove potentially advantageous
in cancer patients bearing exon 14 splice variants of c-Met. As a
result of this apparent restoration of a WT phenotype to a mutant
protein via treatment with PROTAC, we sought to investigate the
PROTAC 7-induced ubiquitination state of c-Met through
immunoprecipitation experiments in the exon 14 mutant cells. After
4 hr of treatment with PROTAC 7, immunoprecipitated c-Met is
ubiquitinated to a greater extent than vehicle control samples
(FIG. 10H).
[1898] Furthermore, Hs746T lysate was subjected to tandem ubiquitin
binding entity 1 (TUBE1) immunoprecipitation in an effort to enrich
for polyubiquitinated substrates within the cell. PROTAC 7-treated
Hs746T cells display marked TUBE1 enrichment of c-Met when compared
with vehicle control-treated cells (FIG. 10I). These experiments
provide evidence that PROTAC 7 induces polyubiquitination of c-Met,
even in an exon 14 skipped context lacking the natural
phosphodegron. While the general applicability of PROTAC-mediated
degradation to RTKs may be inferred beyond the specific examples
described in this study, we are continuing to investigate other
instances of this broad superfamily of proteins as well as the
larger considerations of harnessing the PROTAC approach to the
entire proteome. Additionally, the advantages of inducing
degradation over inhibition of target proteins gleaned from this
"case study" provide a strong foundation for future PROTAC-based
paradigms.
[1899] FIG. 11A-D. FIG. 11A--Quantitative real time PCR was
performed at the indicated timepoints after PROTAC treatment (500
nM). Data is normalized to beta-Tubulin. FIG. 11B-11D
Representative Western blots and quantitation for cotreatment
experiments. FIG. 11B--Co-treatment of PROTAC 7 (500 nM) with
proteasome inhibitor epoxomicin (500 nM) for 6 hours in MDA-MB-231
cells. Quantified data represent average of 2 repeats. FIG.
11C--Co-treatment of PROTAC 7 (500 nM) with neddylation inhibitor
MLN-4924 (1 .mu.M) for 6 hours in MDA-MB-231 cells. Quantified data
represent average of 2 repeats. FIG. 11D--Co-treatment of PROTAC 7
(500 nM) with HSP90 inhibitor 17-AAG (1 .mu.M) for 6 hours in
MDA-MB-231 cells. Quantified data represent average of 2
repeats.
[1900] FIG. 12A-12E. Representative confocal microscopy images of
HGF-mediated internalization of c-Met. FIG. 12A--MDA-MB-231 cells
treated with 100 ng/ml HGF for the indicated times before fixing,
permeabilizing, and immunostaining for c-Met. FIG.
12B--Representative confocal microscopy images demonstrating
PROTAC-mediated colocalization with early endosome antigen 1
(EEA1). MDA-MB-231 cells treated with 500 nM PROTAC 7 for the
indicated times before fixing, permeabilizing, and immunostaining
for c-Met and EEA1. FIG. 12C--Representative confocal microscopy
images demonstrating c-Met co-localization with p230 (a trans-Golgi
marker). FIG. 12D--Quantification of images from FIG. 9C.
Percentage of cellular pixels occupied by c-Met immunofluorescence
and average cellular pixel intensity were used as a proxy for
puncta formation and reduction in cell surface c-Met. FIG.
12E--Clathrin heavy chain (CHC) siRNA experiment. MDA-MB-231 cells
were transfected with CHC siRNA before treatment with PROTAC 7 for
24 hours prior to lysis and immunoblotting.
[1901] Cycloheximide pulse-chase western blots. FIG.
13A--MDA-MB-231 cells were treated with cycloheximide followed by
DMSO, PROTAC 7 or HGF and lysed at the indicated incubation
times--Set 1. FIG. 13B--MDA-MB231 cells were treated with
cycloheximide followed by DMSO, PROTAC 7 or HGF and lysed at the
indicated incubation times--Set 2. FIG. 13C--Hs746T cells were
treated with cycloheximide followed by DMSO, PROTAC 7 or HGF and
lysed at the indicated incubation times. FIG. 13D--c-Met
immunoprecipitation experiments. Hs746T cells were treated with 2
uM epoxomicin for 30 minutes before the addition of PROTAC 7 for 4
hours prior to c-Met immunoprecipitation. (WCL=Whole-cell lysate).
FIG. 13E--Hs746T cells were treated as in D prior to TUBE1
immunoprecipitation experiments.
[1902] Structures of exemplary PROTAC compounds as described herein
(Lapatinib-based (furan) PROTACs) are shown in FIG. 14. Degradation
activity of exemplary PROTAC compounds of FIG. 14. FIGS. 15A and
15B demonstrate the degradation activity of exemplary PROTAC
compounds of FIG. 14. FIG. 15A--the percent degraded HER1 and HER2
protein at 1 uM, linker atoms, linker length (in Angstroms), linker
type and E3 ligase binding moiety (ULM) is indicated. FIG.
15B--demonstrates the degradation activity (dose-response) of HER1
in OVCAR8 cells by lapatinib-based PROTACS as indicated.
[1903] FIGS. 16A and 16B show structures of exemplary lapatinib
(furan)-based PROTACs (FIG. 16A). FIG. 16B--Western blot
demonstrating degradation activity of compounds of FIG. 16A. OVCAR8
treated cells for 24 hours. NRG (5 ng/mL) stimulation for the last
5 minutes. Anti-EGFR rabbit (CST), anti-HER2 (Santa Cruz
Biotechnologies), and anti-tubulin (Sigma-Aldrich) were used for
detection of proteins.
[1904] Degradation activity of exemplary PROTAC compounds. FIG.
17A--shows structures of exemplary lapatinib (phenyl)-based
PROTACs. FIG. 17B--Western blot demonstrating degradation activity
of compounds of FIG. 14A. OVCAR8 treated cells for 24 hours. NRG (5
ng/mL) stimulation for the last 5 minutes. Anti-EGFR rabbit (CST),
anti-HER2 (Santa Cruz Biotechnologies), and anti-tubulin
(Sigma-Aldrich) were used for detection of proteins.
[1905] Tables 1and 2provide C50 and protein degradation data,
respectively, for the indicated exemplary EGFR-PROTACs as described
herein (see FIG. 2). The data demonstrate the ability to inhibit
and degrade wildtype (WT) and mutant forms of EGFR in multiple cell
types, including at clinically relevant concentrations.
TABLE-US-00001 TABLE 1 Time Resolved (TR)-FRET results for
exemplary EGFR PROTACs of Figure 2. Lower IC.sub.50 values (Lower
IC50 values (50% of the maximum inhibition of the corresponding
kinase activity by the test compound) indicate higher affinity for
the corresponding protein. Values in micromolar, geometric mean if
measured more than once. EGFR EGFR EGFR EGFR EGFR L858R/ WT EGFR
A19/746/750 EGFR L858R L858R/ T790M/ HER2 HER3 Ex. IC.sub.50
A19/746/750 T790M Exon20NPG IC.sub.50 T790M C797S HER2WT InsYVMA
IC.sub.50 # (.mu.M) IC.sub.50 (.mu.M) IC.sub.50 (.mu.M) IC.sub.50
(.mu.M) (.mu.M) IC.sub.50 (.mu.M) IC.sub.50 (.mu.M) IC.sub.50
(.mu.M) IC.sub.50 (.mu.M) (.mu.M) 1 0.065 0.0021 6.1 0.23 0.014 12
4.5 1.2 2 0.059 0.0040 7.1 0.51 0.012 >30 3.6 2.3 3 0.21 0.0055
6.6 0.86 0.024 14 14 1.5 4 0.045 0.0042 24 0.29 0.0083 22 10 1.8 5
0.040 0.0041 7.3 0.53 0.011 19 5.4 1.5 6 0.064 0.0029 2.2 0.23
0.047 8.9 3 0.52 7 0.058 0.0010 3.4 0.11 0.040 8.5 2.7 0.30 8 0.089
0.0017 4.8 0.45 0.011 7.2 3.2 1.2 9 0.027 0.0019 3.9 0.26 0.0049
6.8 3.1 0.59 10 1.7 3.7 28 3.2 3.5 16 5.5 3.7 11 0.0084 0.00014 17
0.049 0.00040 >30 3.5 0.19 12 0.50 0.083 8.2 5.7 0.15 9.1 10
>30 13 0.76 0.037 >30 5.3 0.13 >30 >30 >30 14 0.84
0.20 >30 4.4 0.17 >30 7.3 11 15 0.69 0.14 >30 4.7 0.20
>30 17 19 17 0.22 0.19 7.2 4.9 48 0.0047 0.12 3 7 20 66 0.011
0.066 1.2 1.4 6.6 70 >30 >30 11 >30 15 16 >30 19 71 30
>30 3.1 >30 26 0.12 0.096 >30 13 72 >30 22 >30 30
>30 73 20 8.1 6.8 9.8 11 14 >30 3.4 74 >30 >30 >30
10 >30 75 >30 >30 >30 >30 >30 76 17 14 0.36 0.036
8.6 77 24 12 9.1 9.4 >30 78 5.7 5.6 0.25 0.025 25 79 >30
>30 29 7.6 >30 80 4.7 13 4.7 2.2 >30 81 2.4 7.1 8.6 >30
>30 82 3.8 >30 0.77 0.45 >30 83 3 >30 2.5 1.2 >30 84
1.8 5.3 0.68 0.27 >30 85 >30 >30 16 3.9 86 >30 24 12
2.4 87 >30 14 15 3.3 88 >30 3.1 11 9.4 89 >30 >30 1.6
0.65 >30 90 1.3 3.4 0.74 0.69 29 91 4.3 8.3 4.7 0.66 92 9.8 3.1
6.2 1.1 93 11 3.3 3.5 0.53 94 2.1 1.5 6 1.8 95 4.2 3.3 6.7 2.2 96
1.9 2.5 10 4.6 97 1.5 >30 >30 >30 29 101 0.012 0.069 3.4
3.6 103 0.59 0.75 >30 23 104 0.59 0.51 12 8.2 105 0.97 0.82
>30 24 106 0.47 1.9 4.9 5.2 107 0.14 0.6 6.7 6 108 0.77 0.98 10
7.8 109 0.18 0.58 6 8.9 110 0.77 1.8 8.6 12 254 1.1 5.7 1 0.10 255
0.58 4.7 0.83 0.14 256 2.5 6.1 14 1.4 257 5.7 >30 >30 2.7 258
1.7 4.9 5.9 4 259 2.1 2.1 3.1 0.55 260 3.2 >30 20 0.56 261 2.2
4.9 3.5 0.18 262 3.7 7.4 0.43 0.25 263 8.6 12 11 0.79 264 4.2 24
>30 1.1 265 3.6 20 27 0.29 266 10 >30 >30 2.6 267 30 30 30
1.4 275 0.66 9.1 0.88 0.069 276 20 >30 23 29 277 >30 >30
>30 >30 278 30 >30 >30 >30 279 16 >30 29 >30
280 >30 >30 >30 >30 281 12 >30 >30 15 282 12
>30 >30 23 283 1.2 4.6 2.8 0.82 284 14 30 >30 12 285 26 35
17 286 7.3 0.48 0.17 287 6.7 0.074 0.016 288 5.9 6.9 2 289 1.2 1.7
0.19 290 1.1 2.7 0.14 291 14 >60 >60 292 39 19 11 293 1.7 4.2
0.64 294 3.5 0.81 0.17 295 34 3.1 0.71 296 1.6 1.5 0.61 297 1.8 1.8
0.31 298 1.6 1.4 0.54 299 0.24 4.6 0.20 300 2.7 20 3.1 301 2.8 1.6
0.27 0.24 0.034 302 >30 0.67 0.32 303 >30 0.55 0.25 304
>30 0.092 0.017 305 2.1 0.0098 0.0013 306 13 0.38 0.021 0.053
0.016 307 0.37 3.8 1.8 0.71 0.18 308 0.12 0.053 0.0021 0.00091
0.00027 309 0.0072 1.9 5.1 0.12 0.025 310 0.14 6 8.4 0.11 0.022 311
0.097 9 4.6 0.065 0.016 312 0.095 0.17 0.0049 0.00067 0.00035 313
0.0014 0.0075 0.0016 0.00029 0.00026 314 0.089 10 5.2 0.098 0.020
319 0.0042 0.0013 0.0003 0.0023 0.0008
TABLE-US-00002 TABLE 2 Degradation of EGFR protein in various cell
lines. EGFR-wt EGFR EGFR MDA-MB- EGFR L858R/ L858R/ 231 or Exon
T790M T790M/ MCF7 or EGFR- 20 Double C797S A549 or A19 ASV mutant
Triple mutant Ex. # OVCAR8 HCC827 HeLa* H1975 H520* 1 C C C 2 C C C
3 C C C 4 C C C 5 C C C 6 C C C 7 C C 8 C C 9 C C 10 C C 11 C C 12
C C 13 C C 14 C C 15 C C 16 C C 17 C C 36 A C 41 A A B 42 A 43 B B
44 B A 45 A 46 A 47 A A A 48 B A C B 49 A 50 A 51 A A A 56 A 61 A
64 A 66 A B A C 69 A 70 C 71 C 72 C 73 C 74 C 75 C 76 C C 77 C 78 C
C 79 C 80 C C 81 C 82 C 83 C 84 C 85 C 86 C 87 C C 88 C 89 C 90 C C
91 C 92 C C 93 C C 94 C 95 C C 96 C 97 A A B 98 C 99 A 100 C 101 C
C 102 C C 103 C C 104 C C 105 C C 106 C C 107 C C 108 C C 109 C C
110 C C 121 C C 254 C C 255 C B 256 C C 257 C C 258 C C 259 C C 260
C C 261 C C 262 C C 263 C 264 C 265 C C 266 C C 267 C C 268 C 269 C
270 C 271 C 273 C 275 C C B 276 C 277 C 280 C 283 C 284 C 289 C 290
B 291 C 294 C 295 C 296 C 297 B 298 C 299 C 300 C 301 C 302 A 303 A
304 C A 305 C 306 A 307 A 308 B 309 C 310 B 311 B 312 B 313 C A 314
C 319 A 320 A 324 A 325 A 326 A 327 C 328 A 329 C 330 B 331 A 332 A
333 B 334 B 335 B 336 C 338 B 350 C 351 C Degradation of exemplary
compounds (see FIG. 2) data are categorized as follows: A:
.ltoreq.50% EGFR protein remaining after 72 hours of incubation
with the test compound at a concentration between 300 nM and 10 nM;
B: .ltoreq.80% and >50% EGFR protein remaining after 72 hours of
incubation with the test compound at a concentration between 300 nM
and 10 nM; C: >80% EGFR protein remaining after 72 hours of
incubation with the test compound at a concentration of 300 nM.
[1906] In certain embodiments, or in combination with any of the
embodiments described herein, the compounds as described herein
have an IC50 (half maximal inhibitory concentration) for RTK
activity (e.g., EGFR activity) of less than about 1 pM, from about
1 pM to about 1 nM, from about 1 nM to about 1 .mu.M, or from about
1 .mu.M to about 1 mM. In certain additional embodiments, the
compounds as described herein have an IC50 of from about 1 .mu.M to
about 100 .mu.M, from about 10 pM to about 100 .mu.M, or from about
100 pM to about 100 .mu.M. In certain embodiments, the compounds as
described herein have an IC50 of from about 1 .mu.M to about 1
.mu.M, from about 10 pM to about 1 .mu.M, or from about 100 pM to
about 1 .mu.M. In further embodiments, the compounds as described
herein have an IC50 of from about 1 nM to about 1 .mu.M, from about
10 nM to about 1 .mu.M, or from about 100 nM to about 1 .mu.M. In
certain embodiments, the IC50 is determined by TR-FRET method as
described herein.
[1907] In certain additional embodiments, or in combination with
any of the embodiments described herein, the compounds as described
herein exhibit degradation activity of about .ltoreq.50% RTK
protein remaining after 72 hours of incubation with the test
compound at a concentration between about 300 nM and about 10 nM;
from about .ltoreq.80% to about >50% RTK protein remaining after
72 hours of incubation with the test compound at a concentration
between about 300 nM and about 10 nM; or about >80% RTK protein
remaining after 72 hours of incubation with the test compound at a
concentration of 300 nM. In certain embodiments, the degradation is
determined in an in vitro degradation assay as described herein. In
certain embodiments, the in vitro degradation assay is determined
in a cell line selected from OVCAR8, HCC827, HeLa, H1975, or H520
cells.
[1908] As described herein, for the first time it was demonstrated
that PROTACs are capable of inducing the degradation of active
receptor tyrosine kinases and provide examples of successful
degradation of three separate RTKs--EGFR, HER2, and c-Met,
including multiple mutants of EGFR and c-Met. Degradation may
provide advantages over inhibition in several key ways. In most
cases compounds capable of degradation inhibit downstream
signalling and cell proliferation at lower concentrations than
similar compounds that only inhibit. Furthermore, degradation
provides a more sustained reduction in signalling as evidenced by
the reduction in kinome re-wiring observed previously with EGFR,
HER2 and c-Met inhibitors, as well as the sustained duration of
response even after washout. Also, PROTACs are capable of disposing
of proteins that are mutated to avoid their natural "off-switch".
This work significantly expands upon the potential protein targets
of PROTACs to include transmembrane proteins and establishes that
recruitment of VHL to RTKs is capable of efficiently removing this
class of protein targets from the membrane in a similar fashion to
their response to growth factor. Control experiments using the
inactive diastereomeric compounds with identical physicochemical
properties that degradation is leveraged over inhibition alone,
highlighting the potential advantages of this pharmacologic
modality.
[1909] Exemplary Methods
[1910] EGFR Protein Degradation Assay
[1911] HeLa cells or H520 cells stably transfected with flag-tagged
EGFR (exon 20 insert), parental HeLa cells (wild-type EGFR),
NCI-H1975 cells (EGFR: L858R, T790M double-mutant), and A549 cells
(wild-type EGFR) were screened.
[1912] Cells expressing the appropriate form of EGFR were seeded in
6-well plates (300, 000 cells per well) and allowed to adhere to
the plate overnight. Compound (2.times. concentration) was added in
a volume of media equal to that of media in each well to give final
concentrations of 3 .mu.M, 1 .mu.M, 0.3 .mu.M, 0.1 .mu.M, 0.03
.mu.M, together with a DMSO control. Cells were incubated with
compound for 24 hours. To harvest cells, media was removed and
cells were washed once with ice-cold phosphate-buffered saline
(PBS) prior to the addition of 400 .mu.l lysis buffer (Cell
Signaling Technology) supplemented with a protease inhibitor
cocktail (Thermo Scientific). Cells were scraped from the plate and
the cell lysate transferred to an Eppendorf tube and then clarified
by a single spin at 10 000 rpm for 10 minutes. The protein
concentration of each lysate was determined (BCA kit, Thermo
Scientific) prior to the addition of loading buffer (Invitrogen)
and reducing agent (Invitrogen). 10 .mu.g protein of each lysate
was loaded on an SDS/PAGE gel and run at 165V for 2 hours.
Following transfer to a PVDF membrane (iBlot2, Thermo Fisher
Scientific), the membrane was blocked in Tris-buffered saline
containing 0.1% Tween-20 (TBS-T) and supplemented with 5% powdered
milk for 1 hour. Membranes were then probed with EGFR and tubulin
(loaded control) primary antibodies in TBS-T containing 3% bovine
serum albumin (BSA) overnight at 4.degree. C. Membranes were washed
thrice with TBS-T before incubation with secondary antibody in
TBS-T containing 3% bovine serum albumin (BSA) and incubated for 1
hour at room temperature. Following 3 further washes with TBS,
membrane proteins of interest were detected by enzyme-linked
chemiluminescence (ECL) using a Chemidoc (Bio-Rad).
[1913] H520 cells (ATCC: #HTB-182) stably expressing EGFR triple
mutant (L858R, T790M, C797S) were seeded in 1 ml media in 12-well
plates at a density of 100 000 cells per well. Cells were incubated
overnight at 37.degree. C., 5% CO.sub.2 prior to compound addition.
A 10 mM compound stock in DMSO was serially diluted in DMSO to
provide the following stock concentrations: 1, 0.3, 0.1, 0.03 and
0.01 mM. An appropriate volume of each compound dilution, together
with a DMSO vehicle control, was added to media to provide a
2.times. compound solution. 1 ml of each compound solution was then
added to the appropriate well in the 12-well plate to give the
following final compound titrations on cells: DMSO, 1, 0.3, 0.1,
0.03 and 0.01 M. Following compound addition, cells were incubated
for 72 hours at 37.degree. C., 5% CO.sub.2. At the end of the
compound treatment, cells were washed once with ice-cold
phosphate-buffered saline (PBS) and treated with lysis solution (lx
lysis buffer, Cell Signaling Technologies (CST #9803) supplemented
with Piarce.TM. protease inhibitors, #A32953, Thermo Fisher
Scientific, TFS). Lysates were collected and clarified of cell
debris by spinning at 10K rpm for 10 minutes. A specific volume of
supernatant was retrieved for protein quantification (Pierce.TM.
BCA kit, TFS #23225) and for Western analysis. Loading buffer
supplemented with EDTA and reducing agent was added to each
supernatant. A specific volume of supernatant (determined by
protein assay: 1-5 .mu.g protein loaded per sample) was loaded into
a NuPAGE.TM. 4-12% Bis-Tris gel (TFS). Samples were run at 165V in
MOPS running buffer for 2 hours and then transferred to PVDF
blotting membrane using a dry blotting system (iBlot2, TFS).
Membranes were blocked in Tris-buffered saline supplemented with
Tween, 0.1% (TBST) containing 5% non-fat dry milk, (AmericanBio).
Following block, membranes were washed once with TBST. Each
membrane was cut to allow separate staining for EGFR and tubulin
and then appropriate primary antibodies (EGFR, L858R mutant CST,
#3197; .beta.-tubulin, CST #2128) diluted 1:1000 and 1:5000
respectively in TBST with 3% BSA were added and membranes incubated
overnight on a rocker at 4.degree. C. Membranes were washed
3.times. with TBST prior to addition of secondary antibody (1:10
000, anti-rabbit HRP-linked, CST #7074) in TBST with 3% BSA and
then incubated at room temperature for one hour. Membranes were
then washed 3.times. with TBS and signal developed by exposure to
developer (SuperSignal West Femto, TFS) for 5 minutes. Membranes
were then immediately imaged using a BioRad Chemidoc.
[1914] H520 cells stably expressing EGFR triple mutant (L858R,
T790M, C797S) were seeded in 1 ml media in 12-well plates at a
density of 100 000 cells per well. Cells were incubated overnight
at 37.degree. C., 5% CO.sub.2 prior to compound addition. Working
stock concentrations were prepared as for the degradation assay. 1
ml appropriate compound solution or DMSO vehicle control was added
to 1 ml media in the appropriate of well of the 12-well plate.
Cells were incubated for 2 hours at 37.degree. C., 5% CO.sub.2.
Following compound treatment, cells were harvested using methods
identical to those for the degradation assay. Western analysis was
performed exactly as performed for the degradation assay except the
primary antibodies used were phosphoEGFR Y1068 (CST #2234) and
.beta.-tubulin (CST #2128) diluted in TBST with 3% BSA at 1:1000
and 1:5000 dilution respectively.
[1915] Inhibition of Cell Proliferation Assay
[1916] Cells expressing the appropriate form of EGFR were seeded in
96-well plates (2000 cells per well) and allowed to adhere to the
plate overnight. Compound (2.times. concentration) was added in a
volume of media equal to that of media in each well to give a
9-point concentration response curve, with 10 .mu.M top
concentration and diluted 3-fold together with a DMSO control.
Cells were incubated with compound for 72 hours. Cell-titre
Glo.RTM. reagent (Promega) was added to each well and incubated for
30 minutes and the luminescent signal was then read using a
Cytation plate reader (BioTek). Luminescent values for each
compound concentration were normalized to the DMSO vehicle control
and data were plotted and curve fit using GraphPad Prism.
[1917] TR-FRET Assay for ERBB Kinases
[1918] All compounds and PROTACs were serially diluted in
three-fold increments using 100% DMSO, followed by an intermediate
10-fold dilution using Buffer A (50 mM HEPES, pH 7.5, 50 mM NaCl,
10 mM MgCl2, 1 mM DTT, and 0.1% Pluronic F-68). Two microliters of
serially diluted compound or PROTAC were then transferred to black
384-well Proxiplates (PerkinElmer, #6008260) using an Integra
Viaflo96. Next, 10 uL of protein kinase in Buffer A was added to
each well of the assay plate and pre-incubated with compound for 10
minutes. Kinase reactions were then initiated by addition of 5 uL
substrate mix containing 3 mM ATP and 30 uM fluorescein-labeled
Poly-GuTyr (Thermo Fisher, #PV3610) in Buffer A and allowed to
proceed for 10 minutes at room temperature. Reactions were quenched
by addition of a 5 uL mixture containing 5 nM LanthaScreen.RTM.
Tb-pY20 Antibody (Thermo Fisher, #PV3552) and 40 mM EDTA in Buffer
A. Assay plates were then read using a Synergy2 (Biotek
Instruments, Winooski, Vt.) via excitation thru a 340/20 nm
bandpass filter and emission collected thru 490/10 nm (donor) and
520/25 nm (acceptor) bandpass filters. The final kinase
concentrations used for each 15 uL reaction were as follows: 0.2 nM
EGFR Exon20NPG (SignalChem, #E10-132GG), 0.1 nM wild type EGFR (BPS
Bioscience, #40187), 0.3 nM EGFR L858R/T790M/C797S (BPS Bioscience,
#40351), 0.1 nM EGFR L858R (BPS Bioscience, #40189), 0.4 nM
L858R/T790M (BPS Bioscience, #40350), 1 nM EGFR Del19 (SignalChem,
#E10-122JG), 10 nM EGFR Del19 T790M (SignalChem, #E10-122KG), 0.3
nM Her2 (BPS Bioscience, #40230), 15 nM Her2 InsYVMA (SignalChem,
#E27-13BG).
[1919] Experimental Model and Subject Details
[1920] MDA-MB-231, SKBr-3, HCC-827, and H1975 cells were obtained
from the American Type Culture Collection (ATCC). OVCAR8 cells were
a gift from Joyce Liu (Dana Farber). H3255 cells were a gift from
Katerina Politi (Yale University). All of the aforementioned cell
lines were cultured in RPMI-1640 (1.times.) medium containing 10%
fetal bovine serum (FBS) and 1% penicillin-streptomycin and grown
in a humidified incubator at 37.degree. C., 5% CO.sub.2. GTL-16
cells were a gift from F. Mana (Developmental Biology Institute of
Marseille-Luminy) and similarly grown in RPMI-1640 medium
containing 10% FBS and 1% penicillin-streptomycin. To generate an
Exon 20-insertion EGFR stable cell line, HeLa cells (ATCC) were
transduced with a lentiviral mammalian expression vector
pD2119-EFs-3.times.FLAG-EGFR-Exon20ins (purchased from DNA 2.0) and
selected with 2 ug/ml puromycin in Dulbecco's modified Eagle's
medium (DMEM) containing 10% FBS. This vector contains a 767 ASV
duplication of exon 20.
[1921] Immunoblotting.
[1922] Cells were treated with the indicated concentrations of
PROTAC or corresponding inhibiting diastereomer for the specified
time and then harvested in lysis buffer (25 mM Tris-HCl pH 7.5 with
1% NP-40 and 0.25% deoxycholate, supplemented with 10 mM sodium
pyrophosphate, 20 mM -glycerophosphate, 10 mM sodium fluoride, 1 mM
sodium orthovanadate, 0.1 mM phenylarsine oxide, 10 .mu.g/ml
leupeptin, 10 .mu.g/ml pepstatin A, 30 .mu.g/ml bestatin, 0.3
trypsin inhibitor units/ml aprotinin and 1 mM PMSF). Following
centrifugation at 16,000.times.g for 10 min at 4.degree. C. to
pellet insoluble materials, the protein concentrations of the
supernatants were quantitated by BCA assay (Thermo Fisher
Scientific). Protein samples were resolved by 8% SDS-PAGE,
electrophoretically transferred to nitrocellulose and probed with
the antibodies listed above. Immunoblots were developed using
enhanced chemiluminescence and visualized using a Bio-Rad Chemi-Doc
MP Imaging System and quantitated with Image Lab v.5.2.1 software
(Bio-Rad Laboratories).
[1923] Cell Proliferation Assays
[1924] Following PROTAC or diastereomer treatment of cells as
indicated, culture medium was supplemented with 330 .mu.g/ml MTS
(Promega Corp., Madison, Wis.) and 25 .mu.M phenazine methosulfate
(Sigma, St. Louis, Mo.) and incubated at 37.degree. C.
Mitochondrial reduction of MTS to the formazan derivative was
monitored by measuring the medium's absorbance at 490 nm using a
Wallac Victor.sup.2 platereader (Perkin-Elmer Life Sciences,
Waltham, Mass.). Data analysis and statistics performed using Prism
v7.0 software (GraphPad Software).
[1925] Cell Surface Biotinylation Degradation Assay
[1926] A protocol was adapted from Joffre et. al to measure the
removal of c-Met from the cell surface of MDA-MB-231 cells (Joffre
et al., 2011). Cells were plated in full serum, allowed to adhere,
and switched to serum-free RPMI-1640 for 16 hr. After this time,
cells were placed on ice and rinsed with ice-cold 1.times.PBS-CM
(0.1 mM CaCl2, 1 mM MgCl2) twice and incubated with PBS-CM for 5
min at 4.degree. C. PBS-CM was aspirated, at which point cells were
labelled with a cell membrane impermeant reagent, EZ-link
Sulfo-NHS-SS-biotin at 0.5 mg/ml for 30 min at 4.degree. C. with
gentle rocking. This step enabled covalent labelling of all cell
surface proteins. All of the following were carried out at
4.degree. C. to prevent trafficking of said proteins. Cells were
subsequently rinsed with ice-cold PBS-CM twice and excess biotin
was quenched with Tris-glycine buffer (100 mM Tris pH 8.0, 150 mM
NaCl, 0.1 mM CaCl2), 1 mM MgCl2 10 mM glycine, 1% BSA) for 15 min
at 4.degree. C. with gentle rocking. Cells were then rinsed with
ice-cold PBS-CM three times before being chased with warm
serum-free RPMI-1640 medium containing either HGF (100 ng/ml) or
PROTAC (500 nM) and placed in a humidified incubator at 37.degree.
C. for the indicated amount of time, at which point the cells were
lysed with lysis buffer (50 mM Tris, pH 7.5, 100 mM NaCl, 10%
glycerol, 1% NP-40, 1 mM EDTA) supplemented with 1.times. protease
inhibitors (Roche). Lysates were spun down at 14,000.times.g at
4.degree. C. for 10 min and protein content was measured by BCA
assay. Protein lysate was normalized and aliquoted onto
pre-equilibrated NeutrAvidin agarose beads for 2 hrs at 4.degree.
C., with gentle rotation. Beads were washed three times with wash
buffer (100 mM Tris, pH 7.5, 300 mM NaCl, and 1% Triton X-100) and
resuspended in 2.times. elution buffer (62.5 mM Tris, pH 6.8, 3%
SDS, 10% glycerol, 0.02% bromophenol blue, 160 mM DTT). Protein was
eluted off of the beads by heating at 95.degree. C. for 5 min and
the supernatant was run on an SDS-PAGE gel and evaluated for the
presence of cell surface c-Met protein. Whole-cell lysate refers to
the lysate loaded onto NeutrAvidin beads, thereby representing the
total c-Met protein.
[1927] Cycloheximide Chase Assay
[1928] MDA-MB-231 cells were plated at 3.times.10.sup.5 cells per
well in a 6-well dish, allowed to adhere, and switched to
serum-free RPMI-1640 for 16 hr. Cells were then pre-treated with
cycloheximide (Sigma) at 100 ug/ml for 1 hr prior to addition of
either HGF (100 ng/ml), PROTAC (500 nM), or vehicle. At the
indicated time points, cells were immediately placed on ice, rinsed
with PBS, lysed, and boiled.
[1929] Immunofluorescence Microscopy
[1930] MDA-MB-231 cells were plated at a density of
1.times.10.sup.5 cells/ml onto 12 mm round coverslips, cultured
overnight, switched to serum free media for >12 hours and then
treated with 500 nM PROTAC 7 or 100 ng/ml HGF for the indicated
times before washing with PBS. Cells were fixed with 4%
formaldehyde for 20 minutes at room temperature, washed with
ice-cold PBS, permeabilized and blocked with 0.25% Triton X-100/1%
BSA in PBS for 30 minutes. Fixed cells were incubated with c-Met
Antibody (1:3000 dilution, Cell Signalling #8198) for 1 hour,
washed three times with PBS for 5 minutes, incubated with Alexa
Fluor-488 conjugated anti-rabbit antibody (1:1000 dilution,
ThermoFisher A-11008) for 1 hour washed three times with PBS for 5
minutes and mounted in vectashield containing DAPI. Imaged on Zeiss
Axio Observer Z1 inverted microscope.
[1931] siRNA Experiments
[1932] The siRNA (4 .mu.L of 10 .mu.M stock solution, 40 pMol) was
diluted with Opti-MEM media (150 .mu.L) then added to a solution of
Lipofectamine RNAiMAX (9 .mu.L in 150 .mu.L in Opti-MEM) and
incubated for 10 minutes before being added to MDA-MB-231 cells at
.about.80% confluency. The following day, the transfected cells
were plated out and used for experiments as described above.
[1933] Immunoprecipitation Experiments
[1934] Hs746T cells (2.5.times.10.sup.6) were seeded into 10 cm
dishes, allowed to adhere, switched to serum-free DMEM media for 16
hr. After this time, cells were pre-treated with 2 uM epoxomicin
for 30 minutes at 37.degree. C. After this pre-treatment, 10 cm
plates were treated with either 1 uM Compound 7 or vehicle for 4
hours at 37.degree. C., after which they were placed on ice, rinsed
twice with ice-cold 1.times.PBS and lysed with 500 uL modified
1.times.RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40,
1% sodium deoxycholate, 0.1% SDS) containing 5 mM
1,10-phenanthroline monohydrate, 10 mM N-ethylmaleimide, 20 uM
PR-619, and 1.times. complete protease inhibitor cocktail (Roche).
Lysates were spun down at 14,000.times.g at 4.degree. C. for 10 min
and protein content was measured by BCA assay. Protein lysate was
normalized and 500 ug of lysate was aliquoted onto naked Protein
A-Sepharose 4B beads (Sigma), and pre-cleared for 1 hr at 4.degree.
C. with gentle rotation. After this 1 hr incubation, samples were
spun down at 3,000.times.g at 4.degree. C. for 2 min and the
normalized, pre-cleared lysate were subsequently loaded onto
Protein A-Sepharose 4B beads coupled with 5 ug of Met (CST, #8198)
antibody. MET was immunoprecipitated from Hs746T lysates for 2 hr
at 4.degree. C. with gentle rotation, after which samples were spun
down at 3,000.times.g at 4.degree. C. for 2 min, flow-thru was
collected to assess pulldown efficiency (see FIG. S6D), and the
beads were washed once with ice-cold lysis buffer and thrice with
ice-cold 1.times.TBS-T (137 mM NaCl, 2.7 mM KCl, 19 mM Tris-HCl pH
7.5, 0.02% Tween-20). The beads were resuspended in 1.times.LDS
sample buffer containing 5% BME. Immunoprecipitated protein was
eluted off of the beads by heating at 95.degree. C. for 5 min and
the supernatant was run on an SDS-PAGE gel and evaluated for the
presence of immunoprecipitated total Met (CST, #3127), as well as
ubiquitinated Met (CST, #3936). Whole-cell lysate refers to the
normalized, input lysate loaded onto Protein A-Sepharose beads.
[1935] TUBE1 Immunoprecipitation Experiments
[1936] TUBE1 immunoprecipitations were carried out exactly as
described in the previous section (Immunoprecipitation
Experiments), except for the fact that 1 mg of Hs746T lysate was
used and loaded onto 20 uL TUBE1 agarose (LifeSensors) resin per
sample.
EMBODIMENTS OF THE PRESENT DISCLOSURE
[1937] The present disclosure encompasses the following specific
embodiments. These following embodiments may include all of the
features recited in a proceeding embodiment, as specified. Where
applicable, the following embodiments may also include the features
recited in any proceeding embodiment inclusively or in the
alternative.
[1938] In certain aspects, the description provides a bifunctional
compound having the chemical structure: PTM-Linker-ULM, or a
pharmaceutically acceptable salt, enantiomer, stereoisomer,
solvate, polymorph or prodrug thereof, wherein: ULM is a small
molecule E3 ubiquitin ligase binding moiety that binds an E3
ubiquitin ligase; PTM is a small molecule receptor tyrosine kinase
(RTK) protein targeting moiety, wherein the PTM is at least one of;
and Linker (L) is a bond or a chemical linking moiety covalently
coupling the ULM and the PTM.
[1939] In any of the aspects or embodiments described herein, the
ULM is a moiety that binds an E3 ligase protein selected from the
group consisting of Von Hippel-Lindau, cereblon, mouse
double-minute homolog2, and IAP as described and exemplified
herein. In any of the aspects or embodiments described herein, the
described compounds include a PTM that comprises the structure of
any of formulas I-XVII as described herein, including all
variations described. In any of the aspects or embodiments
described herein, the PTM is coupled to the ULM via a linker,
wherein the linker (L) is a bond or chemical linker moiety as
described herein. In any of the aspects or embodiments described
herein, the linker is coupled to the PTM via an R group as
described for formulas I-XVII.
[1940] In any of the aspects or embodiments described herein, the
compound includes a linker (L) moiety having a structure as
described herein coupling the PTM to the ULM. For example, in any
of the aspects or embodiments, the linker comprises a chemical
structural unit represented by the formula: -(A.sup.L)q-, wherein:
(A.sup.L)q is a group which is connected to at least one of a ULM,
a PTM moiety, or a combination thereof; q is an integer greater
than or equal to 1; each A.sup.L is independently selected from the
group consisting of, a bond, CRL1RL2, O, S, SO, SO2, NRL3, SO2NRL3,
SONRL3, CONRL3, NRL3CONRL4, NRL3SO2NRL4, CO, CRL1=CRL2, C.ident.C,
SiRL1RL2, P(O)RL1, P(O)ORL1, NRL3C(.dbd.NCN)NRL4, NRL3C(.dbd.NCN),
NRL3C(.dbd.CNO2)NRL4, C3-11cycloalkyl optionally substituted with
0-6 RL1 and/or RL2 groups, C3-11heteocyclyl optionally substituted
with 0-6 RL1 and/or RL2 groups, aryl optionally substituted with
0-6 RL1 and/or RL2 groups, heteroaryl optionally substituted with
0-6 RL1 and/or RL2 groups, where RL1 or RL2, each independently are
optionally linked to other groups to form cycloalkyl and/or
heterocyclyl moiety, optionally substituted with 0-4 RL5 groups;
and RL1, RL2, RL3, RL4 and RL5 are, each independently, H, halo,
C1-8alkyl, OC1-8alkyl, SC1-8alkyl, NHC1-8alkyl, N(C1-8alkyl)2,
C3-11cycloalkyl, aryl, heteroaryl, C3-11heterocyclyl,
OC1-8cycloalkyl, SC1-8cycloalkyl, NHC1-8cycloalkyl,
N(C1-8cycloalkyl)2, N(C1-8cycloalkyl)(C1-8alkyl), OH, NH2, SH,
SO2C1-8alkyl, P(O)(OC1-8alkyl)(C1-8alkyl), P(O)(OC1-8alkyl)2,
CC-C1-8alkyl, CCH, CH.dbd.CH(C1-8alkyl),
C(C1-8alkyl).dbd.CH(C1-8alkyl), C(C1-8alkyl).dbd.C(C1-8alkyl)2,
Si(OH)3, Si(C1-8alkyl)3, Si(OH)(C1-8alkyl)2, COC1-8alkyl, CO2H,
halogen, CN, CF3, CHF2, CH2F, NO2, SF5, SO2NHC1-8alkyl,
SO2N(C1-8alkyl)2, SONHC1-8alkyl, SON(C1-8alkyl)2, CONHC1-8alkyl,
CON(C1-8alkyl)2, N(C1-8alkyl)CONH(C1-8alkyl),
N(C1-8alkyl)CON(C1-8alkyl)2, NHCONH(C1-8alkyl), NHCON(C1-8alkyl)2,
NHCONH2, N(C1-8alkyl)SO2NH(C1-8alkyl), N(C1-8alkyl)
SO2N(C1-8alkyl)2, NH SO2NH(C1-8alkyl), NH SO2N(C1-8alkyl)2, NH
SO2NH2.
[1941] In any of the aspects or embodiments, the linker (L) has a
chemical structure selected from:
##STR00812##
wherein W.sup.L1 and W.sup.L2 are each independently a 4-8 membered
ring with 0-4 heteroatoms, optionally substituted with R.sup.Q,
each R.sup.Q is independently a H, halo, OH, CN, CF.sub.3,
C.sub.1-C.sub.6 alkyl (linear, branched, optionally substituted),
C.sub.1-C.sub.6 alkoxy (linear, branched, optionally substituted),
or 2 R.sup.Q groups taken together with the atom they are attached
to, form a 4-8 membered ring system containing 0-4 heteroatoms;
Y.sup.L1 is each independently a bond, C.sub.1-C.sub.6 alkyl
(linear, branched, optionally substituted) and optionally one or
more C atoms are replaced with O; or C.sub.1-C.sub.6alkoxy (linear,
branched, optionally substituted); n is 0-10; and a dashed line
indicates the attachment point to the PTM or ULM moieties.
[1942] In any of the aspects or embodiments described herein, the
compound comprises a linker (L) which is a polyethylenoxy group
optionally substituted with aryl or phenyl comprising from 1 to 10
ethylene glycol units.
[1943] In any of the aspects or embodiments described herein, the
compound has the structure selected from compounds 1-351 (FIG. 2),
including analogs, derivatives, salts, prodrugs, polymorphs,
analogs, derivatives, and deuterated forms thereof. In an
additional aspect, the description provides a therapeutic
composition comprising an effective amount of at least one of
compounds 1-351 (FIG. 2), and pharmaceutically acceptable carrier.
In any of the aspects or embodiments described herein, the
composition further comprises at least one of an additional
bioactive agent or another PROTAC compound as described and/or
exemplified herein. In any of the aspects or embodiments, the
additional bioactive agent is an anti-cancer (i.e., anti-oncologic)
or anti-inflammatory agent. In any of the aspects or embodiments,
the PROTAC compounds as described herein are co-administered
(together or separately) with an anti-oncologic agent. In any of
the aspects or embodiments described herein, the anti-oncologic is
an anti-PD1 or anti-PD-L1 antibody. In an aspect, the description
provides a therapeutic composition for co-administration comprising
an effective amount of a compound as described herein, and an
effective amount additional biologically active agent, for example,
an anti-oncologic agent. In certain embodiments, the effective
amount of the compounds as described herein, and the effective
amount of the additional biologically active agent are comprised in
separate containers.
[1944] In an aspect, the description provides a composition
comprising a pharmaceutically acceptable carrier and an effective
amount of at least one compound as described herein for treating a
disease or disorder in a subject. In any of the aspects or
embodiments, the disease or disorder is associated with receptor
tyrosine kinase, e.g., EGFR, c-MET, HER1-3, or VEGFR,
overexpression or hyperactivity. In any of the aspects or
embodiments, the disease or disorder is cancer. In any of the
aspects or embodiments, the disease or disorder is at least one of
squamous-cell carcinoma of the lung, colon and anal cancers,
glioblastoma, and epithelial tumors of the head and neck,
psoriasis, eczema and atherosclerosis or a combination thereof.
[1945] In additional aspects, the description provides methods of
treating a receptor tyrosine kinase (RTK)-related disease or
disorder in a subject comprising administering to a subject in need
thereof an effective amount of at least one compound as described
herein, or a therapeutic composition comprising the same, wherein
the at least one compound or composition is effective for
ameliorating at least one symptom of the RTK-related disease or
disorder. In any of the aspects or embodiments described herein,
the RTK-related disease or disorder is cancer, inflammatory disease
or reduced hair growth, that is associated with RTK overexpression
or hyper-activity. In any of the aspects or embodiments described
herein, the disease or disorder is related to EGFR overexpression
or hyper-activity.
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