U.S. patent application number 15/206497 was filed with the patent office on 2017-01-12 for mdm2-based modulators of proteolysis and associated methods of use.
The applicant listed for this patent is Arvinas, Inc.. Invention is credited to Andrew P. Crew, Craig M. Crews, Hanqing Dong, Yimin Qian, Jing Wang.
Application Number | 20170008904 15/206497 |
Document ID | / |
Family ID | 57730930 |
Filed Date | 2017-01-12 |
United States Patent
Application |
20170008904 |
Kind Code |
A1 |
Crew; Andrew P. ; et
al. |
January 12, 2017 |
MDM2-BASED MODULATORS OF PROTEOLYSIS AND ASSOCIATED METHODS OF
USE
Abstract
The description relates to MDM2 binding compounds, including
bifunctional compounds comprising the same, which find utility as
modulators of targeted ubiquitination, especially inhibitors of a
variety of polypeptides and other proteins which are degraded
and/or otherwise inhibited by bifunctional compounds according to
the present invention. In particular, the description provides
compounds, which contain on one end a ligand which binds to the
MDM2 E3 ubiquitin ligase and on the other end a moiety which binds
a target protein such that the target protein is placed in
proximity to the ubiquitin ligase to effect degradation (and
inhibition) of that protein. Compounds can be synthesized that
exhibit a broad range of pharmacological activities consistent with
the degradation/inhibition of targeted polypeptides of nearly any
type.
Inventors: |
Crew; Andrew P.; (Guilford,
CT) ; Crews; Craig M.; (New Haven, CT) ; Dong;
Hanqing; (Madison, CT) ; Qian; Yimin;
(Plainsboro, NJ) ; Wang; Jing; (Milford,
CT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Arvinas, Inc. |
New Haven |
CT |
US |
|
|
Family ID: |
57730930 |
Appl. No.: |
15/206497 |
Filed: |
July 11, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62191193 |
Jul 10, 2015 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 35/00 20180101;
A61K 31/337 20130101; A61K 31/4184 20130101; A61K 31/4545 20130101;
A61K 31/436 20130101; C07D 401/14 20130101; A61K 47/55 20170801;
A61K 45/06 20130101; C07D 207/16 20130101; A61K 31/4995 20130101;
C07D 405/14 20130101; C07D 401/12 20130101; A61K 31/506 20130101;
C07D 417/12 20130101; A61K 31/704 20130101; C07D 495/14 20130101;
A61K 31/593 20130101; A61K 31/664 20130101; A61P 35/02 20180101;
A61P 43/00 20180101; A61K 31/337 20130101; A61K 2300/00 20130101;
A61K 31/436 20130101; A61K 2300/00 20130101; A61K 31/4184 20130101;
A61K 2300/00 20130101; A61K 31/4545 20130101; A61K 2300/00
20130101; A61K 31/506 20130101; A61K 2300/00 20130101; A61K 31/593
20130101; A61K 2300/00 20130101; A61K 31/664 20130101; A61K 2300/00
20130101; A61K 31/704 20130101; A61K 2300/00 20130101; A61K 31/4995
20130101; A61K 2300/00 20130101 |
International
Class: |
C07D 495/14 20060101
C07D495/14; C07D 417/12 20060101 C07D417/12; C07D 401/14 20060101
C07D401/14; C07D 207/16 20060101 C07D207/16; C07D 401/12 20060101
C07D401/12; C07D 405/14 20060101 C07D405/14; A61K 47/48 20060101
A61K047/48; A61K 31/551 20060101 A61K031/551; A61K 31/40 20060101
A61K031/40; A61K 31/427 20060101 A61K031/427; A61K 31/4166 20060101
A61K031/4166; A61K 31/496 20060101 A61K031/496; A61K 31/166
20060101 A61K031/166; A61K 31/4433 20060101 A61K031/4433; A61K
31/506 20060101 A61K031/506; A61K 45/06 20060101 A61K045/06 |
Claims
1. A compound having a chemical structure comprising of: PTM-L-MLM
wherein MLM is a MDM2 E3 ubiquitin ligase binding moiety, PTM is a
protein targeting moiety, and L is a linker coupling the MLM to the
PTM, and wherein the PTM binds to a targeted protein having a
function or activity selected from the group consisting of:
structural protein, regulatory, growth factor, receptor,
cytoskeletal, hormonal, enzymatic, nucleic acid binding,
immunological, contractile, storage, transportation, signal
transduction, catalytic activity, protein binding, aromatase
activity, lipase, protease, nuclease, motor activity, helicase
activity, metabolism, antioxidant activity, proteolysis,
biosynthesis, kinase, oxidoreductase, transferase, hydrolase,
lyase, isomerase, ligase, enzyme regulator, signal transducer,
protein or lipid binding, cell motility, membrane fusion, cell
communication, cell growth or differentiation, cell division,
response to stimulus, cell adhesion, apoptosis, transport,
secretion, electron transport, ion channel, chaperone or chaperone
regulator, nucleic acid binding activity, transcription regulator,
extracellular organization and biogenesis, and translation
regulator, or a pharmaceutically acceptable salt, enantiomer,
stereoisomer, solvate, polymorph or prodrug thereof.
2. The compound of claim 1, wherein the PTM binds to a protein
selected from the group consisting of B7.1, B7, TINFR1m, TNFR2,
NADPH oxidase, Bcl, Bax, apotosis pathway proteins, C5a receptor,
HMG-CoA reductase, PDE V phosphodiesterase, PDE IV
phosphodiesterase, PDE I, PDEII, PDEIII, squalene cyclase
inhibitor, CXCR1, CXCR2, nitric oxide (NO) synthase,
cyclo-oxygenase 1, cyclo-oxygenase 2, 5HT receptor, dopamine
receptor, G Protein, Gq, histamine receptor, 5-lipoxygenase,
tryptase serine protease, thymidylate synthase, purine nucleoside
phosphorylase, GAPDH trypanosomal, glycogen phosphorylase, carbonic
anhydrase, chemokine receptor, JAK, STAT, RXR, HIV 1 protease, HIV
1 integrase, influenza, neuramimidase, hepatitis B reverse
transcriptase, sodium channel, multi drug resistance (MDR), protein
P-glycoprotein (and MRP), tyrosine kinase, CD23, CD124, tyrosine
kinase p56 lck, CD4, CD5, IL-2 receptor, IL-1 receptor, TNF-alphaR,
ICAM1, Cat+ channel, VCAM, VLA-4 integrin, selectin, CD40/CD40L,
receptor, inosine monophosphate dehydrogenase, p38 MAP Kinase, JNK,
Ras, Raf, ERK, FLT-3, KSR1, SMARCA, SMARCA2, interleukin-1
converting enzyme, caspase, HCV, NS3 protease, HCV NS3 RNA
helicase, glycinamide ribonucleotide formyl transferase, rhinovirus
3C protease, herpes simplex virus-1 (HSV-I), protease,
cytomegalovirus (CMV) protease, poly (ADP-ribose) polymerase,
cyclin dependent kinase, growth factor, growth factor receptor,
receptor tyrosine kinase, cytokine, GPCR, vascular endothelial
growth factor, EGF, EGFR, HGF, HGFR, VEGF, VEGFR, Wnt, TNF-.alpha.,
TPO, TCGF, PGF, NT-3, NT-4, TGF, TGF-.beta., oxytocin receptor,
microsomal transfer protein inhibitor, bile acid transport
inhibitor, 5 alpha reductase, angiotensin 11, glycine receptor,
noradrenaline reuptake receptor, endothelin receptor, neuropeptide
Y and receptor, estrogen receptor, androgen receptor, adenosine
receptor, adenosine kinase and AMP deaminase, purinergic receptor,
P2Y1, P2Y2, P2Y4, P2Y6, P2X1-7, an E1, E2 or E3 ubiquitin ligase,
VHL, cereblon, p53, farnesyltransferase, geranylgeranyl
transferase, TrkA a receptor for NGF, beta-amyloid, tyrosine kinase
Flk-IIKDR, vitronectin receptor, integrin receptor, Her-21 neu,
telomerase, cytosolic phospholipaseA2 and EGF receptor tyrosine
kinase, ecdysone 20-monooxygenase, GABA gated chloride channel,
acetylcholinesterase, voltage-sensitive sodium channel protein,
calcium release channel, chloride channel, Acetyl-CoA carboxylase,
adenylosuccinate synthetase, protoporphyrinogen oxidase,
enolpyruvylshikimate-phosphate synthase, haloalkane halogenase
inhibitors, Hsp90, kinase, MDM2, human BET Bromodomain-containing
protein, HDAC, EZH2, human lysine methyltransferase, and aryl
hydrocarbon receptor (AHR).
3. The compound of claim 1, wherein the PTM binds to a protein
selected from the group consisting of a kinases, enzymes,
transporters, nuclear hormone receptors, non-nuclear hormone
receptors, G-protein coupled receptors (GPCRs), transcription
factors, and epigenetic targets particularly, a human BET
Bromodomain-containing protein (BRD), Brd4, Ras, Raf, MDM2,
androgen receptor (AR) and estrogen receptor (ER), EZH2 and
JNK.
4. A compound having a structure selected from the group consisting
of: ##STR00126## wherein, PTM is a protein targeting moiety that
binds a target protein, and L is a linker coupling the PTM to the
molecule shown; X is selected from the group consisting of carbon,
oxygen, sulfur, sulfoxide, sulfone, and N--R.sup.a; R.sup.a is
independently H or an alkyl group with carbon number 1 to 6; Y and
Z are independently carbon or nitrogen; A, A' and A'' are
independently selected from C, N, O or S, can also be one or two
atoms forming a fused bycyclic ring, or a 6,5- and 5,5-fused
aromatic bicyclic group; R.sub.1, R.sub.2 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: 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 are independently selected from the group
consisting of H, methyl and C1 to C6 alkyl; R.sub.5 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: 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 is H or
--C(.dbd.O)R.sup.b, wherein R.sup.b 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 R.sup.c 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 R.sup.d 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 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.sub.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: R.sup.e is an alkylene with 1 to 6 carbons, or a bond;
R.sup.f is a substituted 4- to 7-membered heterocycle; R.sup.g is
selected from the group consisting of aryl, hetero aryl,
substituted aryl or heteroaryl, and 4- to 7-membered heterocycle;
R.sub.9 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 consistin of halogen, alkene, alkyne, alkyl,
unsubstituted or substituted with Cl or F; R.sub.10 is selected
from the group consistin 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 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: 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 R.sup.h and
R.sup.i 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 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
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.sub.15 is CN; R.sub.16 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(allkyl), --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 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, hydrxylated 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, C1-6 alkyl with hydrogen replaced by a cyano group, 5
and 6 membered aryl or heteroaryl, aklyl 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 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 is
selected from the group consisting of aryl, heteroaryl, bicyclic
heteroaryl, and these aryl or hetroaryl 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 are independently selected
from C1-6 alkyl, C1-6 cycloalkyl, C1-6 alkoxy, hydoxylated 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 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 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, hydoxylated C1-6 alkyl,
cycloalkyl, fluorine-substituted C1-6 alkyl, CN, alkoxy, alkyl
sulfone, amide and sulfonamide; R.sub.24 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, hydoxylated alkyl,
cyano-substituted alkyl, cycloalyl and substituted cycloalky;
R.sub.25 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 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 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 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'' is
selected from the group consisting of alkyl, aryl substituted
alkyl, alkoxy substituted alkyl, cycloalkyl, aryl-substituted
cycloalkyl, and alkoxy substituted cycloalkyl, or a
pharmaceutically acceptable salt, enantiomer, stereoisomer,
solvate, polymorph or prodrug thereof.
5. The compound of claim 4, wherein the heterocycles in R.sup.f and
R.sup.g are independently selected from the group consisting of
substituted pyrrolidine, substituted piperidine, and substituted
piperizine.
6. The compound of claim 4, wherein the R.sub.9 substituents are
selected from Cl and F.
7. The compound of claim 4, wherein the R.sub.10 substituents are
selected from H, F and Cl.
8. The compound of claim 4, wherein R.sup.h and R.sup.i are
selected from the group consisting of: (i) R.sup.h is H, and
R.sup.i is 4-hydroxycyclohehexane; (ii) R.sup.h is H, and R.sup.i
is mono- and di-hydroxy substituted lower alkyl (C3 to C6); (iii)
R.sup.h is H, and R.sup.i is 3-hydroxycyclobutane; and (iv) R.sup.h
is H, and R.sup.i is phenyl-4-carboxylic acid, substituted
phenyl-4-carboxylic acid.
9. The compound of claim 4, wherein the R.sub.18 substitution is
selected from the group consisting of --N(C1-4 alkyl)(cycloalkyl),
--N(C1-4 alkyl)alkyl-cycloalkyl, and --N(C1-4 alkyl)
[(alkyl)-(heterocycle-substituted)-cycloalkyl].
10. The compound of claim 4, wherein the R.sub.28 saturated
heterocycle is selected from piperidine, piperidinone,
tetrahydropyran, and N-acyl-piperidine.
11. The compound of claim 4, wherein the compound has a structure
selected from the group consisting of: ##STR00127## wherein R1' and
R2' 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' and R6' are independently 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; and R5' 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, or a
pharmaceutically acceptable salt, enantiomer, stereoisomer,
solvate, polymorph or prodrug thereof.
12. The compound of claim 11, wherein the linker is attached to at
least one of R1', R2', R3', R4', R5', R6', or a combination
thereof.
13. The compound of claim 11, wherein R6' is independently selected
from the group consisting of H, ##STR00128## ##STR00129##
##STR00130## ##STR00131## ##STR00132## ##STR00133## ##STR00134##
wherein * indicates the point of attachment of the linker.
14. The compound of claim 4, wherein the compound has a structure
selected from the group consisting of: ##STR00135## ##STR00136##
wherein R.sub.7' is a member selected from the group consisting of
halogen, mono-, and di- or tri-substituted halogen; R.sub.8' 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;
R.sub.9' is selected from the group consistin 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 is selected from the group consistin of H,
--OCH.sub.3, --OCH.sub.2CH.sub.3, and halogen; R.sub.10' and
R.sub.11' 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).sup.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)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).sub.2, oxo, carboxy, clcloalkyl and heteroaryl; m, n, and
p are independently 0 to 6; R.sub.12' is selected from the group
consisting of --O-(alkyl), --O-(alkyl)-akoxy, --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); and R.sub.1'' is selected from
the group consisting of alkyl, aryl substituted alkyl, aloxy
substituted alkyl, cycloalkyl, ary-substituted cycloalkyl, and
alkoxy substituted cycloalkyl, or a pharmaceutically acceptable
salt, enantiomer, stereoisomer, solvate, polymorph or prodrug
thereof.
15. The compound of claim 14, wherein the linker is attached to at
least one of Z, R.sub.8', R.sub.9', R.sub.10', R.sub.11',
R.sub.12', R.sub.1'', or a combination thereof.
16. The compound of any of claim 4, 11 or 14, wherein the linker
group (L) comprises a chemical structural unit represented by the
formula: -Aq- wherein: q is an integer greater than 1; and A 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; wherein RL1, RL2, RL3, RL4 and RL5 are each, independently,
selected from the group consisting of 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, and NH
SO2NH2; and wherein: when q is greater than 1, RL1 or RL2 each,
independently, can be linked to another A group to form cycloalkyl
and/or heterocyclyl moeity that can be further substituted with 0-4
RL5 groups.
17. The compound of claim 16 wherein the linker group (L) is
selected from the structure consisting of: ##STR00137##
##STR00138## ##STR00139## ##STR00140## ##STR00141## ##STR00142##
wherein: "X" is a linear chain with atoms ranging from 2 to 14 with
heteroatoms optionally; and "Y" is O, N and S(O).sub.n wherein,
(n=0, 1, 2).
18. The compound of claim 1, wherein the PTM group is a protein
target moiety that binds to bromodomain-containing protein 4
(BRD4).
19. The compound of claim 18, wherein the PTM is a protein target
moiety that binds to a human BET Bromodomain-containing protein is
selected from the structure consisting of: ##STR00143##
##STR00144## wherein * indicates the point of attachment of the
linker.
20. The compound of claim 1, wherein the PTM is a protein target
moiety selected from the structure consisting of: ##STR00145##
wherein * indicates the point of attachment of the linker.
21. The compound according to claim 1, wherein the MLM comprises
part of structural feature as in at least one of RG7112, RG7388,
SAR405838, AMG-232, AM-7209, DS-5272, MK-8242, or NVP-CGM-097, and
analogs or derivatives thereof.
22. A compound is selected from the group consisting of chemical
formula:
4-(3-{4-[2-(2-{4-[2-(4-tert-butyl-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)-
-4,5-dimethyl-4,5-dihydro-1H-imidazole-1-carbonyl]piperazin-1-yl}ethoxy)et-
hoxy]phenyl}-4,4-dimethyl-5-oxo-2-sulfanylideneimidazolidin-1-yl)-2-(trifl-
uoromethyl)benzonitrile;
4-(3-{4-[(17-{4-[2-(4-tert-butyl-2-ethoxyphenyl)-4,5-bis
(4-chlorophenyl)-4,5-dimethyl-4,5-dihydro-1H-imidazole-1-carbonyl]piperaz-
in-1-yl}-3,6,9,12,15-pentaoxaheptadecan-1-yl)oxy]phenyl}-4,4-dimethyl-5-ox-
o-2-sulfanylideneimidazolidin-1-yl)-2-(trifluoromethyl)benzonitrile;
N-(17-{[3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-
-(2,2-dimethylpropyl)pyrrolidin-2-yl]formamido}-3,6,9,12,15-pentaoxaheptad-
ecan-1-yl)-2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-te-
traazatricyclo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetami-
de;
N-(2-{2-[2-(2-{[3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl-
)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidin-2-yl]formamido}ethoxy)ethoxy]et-
hoxy}ethyl)-2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-t-
etraazatricyclo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetam-
ide;
N-(14-{[3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cya-
no-5-(2,2-dimethylpropyl)pyrrolidin-2-yl]formamido}-3,6,9,12-tetraoxatetra-
decan-1-yl)-2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-t-
etraazatricyclo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetam-
ide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,-
2-dimethylpropyl)-N-{2-[2-(4-{[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2-
,4,4-tetramethylcyclobutyl]carbamoyl}phenoxy)ethoxy]ethyl}pyrrolidine-2-ca-
rboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-
-5-(2,2-dimethylpropyl)-N-[1-(4-{[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)--
2,2,4,4-tetramethylcyclobutyl]carbamoyl}phenyl)-1,4,7,10-tetraoxadodecan-1-
2-yl]pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-(2-{2-[2-(4-{[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,-
4,4-tetramethylcyclobutyl]carbamoyl}phenoxy)ethoxy]ethoxy}ethyl)pyrrolidin-
e-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-[1-(4-{[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-te-
tramethylcyclobutyl]-carbamoyl}phenyl)-1,4,7,10,13,16-hexaoxaoctadecan-18--
yl]pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-[1-(4-{[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-te-
tramethylcyclobutyl]carbamoyl}phenyl)-1,4,7,10,13-pentaoxapentadecan-15-yl-
]pyrrolidine-2-carboxamide;
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-[1-(4-{[(1,3-trans)-3-(3-chloro-4-cyanopheno-
xy)-2,2,4,4-tetramethylcyclobutyl]carbamoyl}phenyl)-1,4,7,10,13-pentaoxape-
ntadecan-15-yl]pyrrolidine-2-carboxamide;
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-[1-(4-{[(1r,3r)-3-(3-chloro-4-cyanophenoxy)--
2,2,4,4-tetramethylcyclobutyl]carbamoyl}phenyl)-1,4,7,10,13-pentaoxapentad-
ecan-15-yl]pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-{4-[(2-{2-[2-(2-
-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetraazatri-
cyclo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}ethoxy-
)ethoxy]ethoxy}ethyl)carbamoyl]-2-methoxyphenyl}-4-cyano-5-(2,2-dimethylpr-
opyl)pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-[4-({2-[2-(2-{2-
-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetraazatricyc-
lo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}ethoxy)et-
hoxy]ethyl}carbamoyl)-2-methoxyphenyl]-4-cyano-5-(2,2-dimethylpropyl)pyrro-
lidine-2-carboxamide;
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-[-
4-({2-[2-(2-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12--
tetraazatricyclo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]aceta-
mido}ethoxy)ethoxy]ethyl}carbamoyl)-2-methoxyphenyl]-4-cyano-5-(2,2-dimeth-
ylpropyl)pyrrolidine-2-carboxamide;
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-[-
4-({2-[2-(2-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12--
tetraazatricyclo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]aceta-
mido}ethoxy)ethoxy]ethyl}carbamoyl)-2-methoxyphenyl]-4-cyano-5-(2,2-dimeth-
ylpropyl)pyrrolidine-2-carboxamide;
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-{-
4-[(2-{2-[2-(2-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,-
12-tetraazatricyclo
[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}ethoxy)eth-
oxy]ethoxy}ethyl)
carbamoyl]-2-methoxyphenyl}-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-c-
arboxamide;
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-{-
4-[(2-{2-[2-(2-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,-
12-tetraazatricyclo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]ac-
etamido}ethoxy)
ethoxy]ethoxy}ethyl)carbamoyl]-2-methoxyphenyl}-4-cyano-5-(2,2-dimethylpr-
opyl)pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-{4-[(14-{2-[(9S-
)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetraazatricyclo[8.-
3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}-3,6,9,12-tetr-
aoxatetradecan-1-yl)carbamoyl]-2-methoxyphenyl}-4-cyano-5-(2,2-dimethylpro-
pyl)pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-[4-({2-[2-({[(2S)-1-[(2S,4R)-4-hydroxy-2-({[4-(4-methyl-1,-
3-thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxo-
butan-2-yl]carbamoyl}methoxy)ethoxy]ethyl}carbamoyl)-2-methoxyphenyl]pyrro-
lidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-(4-{[2-(2-{2-[(-
9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetraazatricyclo[-
8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}ethoxy)ethyl-
]carbamoyl}-2-methoxyphenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-c-
arboxamide;
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-(-
4-{[2-(2-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tet-
raazatricyclo
[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}ethoxy)eth-
yl]carbamoyl}-2-methoxyphenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-
-carboxamide;
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-(-
4-{[2-(2-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tet-
raazatricyclo
[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}ethoxy)eth-
yl]carbamoyl}-2-methoxyphenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-
-carboxamide;
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-{-
4-[(14-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetra-
azatricyclo
[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}-3,6,9,12--
tetraoxatetradecan-1-yl)carbamoyl]-2-methoxyphenyl}-4-cyano-5-(2,2-dimethy-
lpropyl)pyrrolidine-2-carboxamide;
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-{-
4-[(14-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetra-
azatricyclo
[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}-3,6,9,12--
tetraoxatetradecan-1-yl)carbamoyl]-2-methoxyphenyl}-4-cyano-5-(2,2-dimethy-
lpropyl)pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-{4-[(1-{[(2S)-1-[(2S,4R)-4-hydroxy-2-({[4-(4-methyl-1,3-th-
iazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobuta-
n-2-yl]carbamoyl}-2,5,8,11-tetraoxatridecan-13-yl)carbamoyl]-2-methoxyphen-
yl} pyrrolidine-2-carboxamide;
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-{4-[(1-{[(2S)-1-[(2S,4R)-4-hydroxy-2-({[4-(4-
-methyl-1,3-thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3,3-dime-
thyl-1-oxobutan-2-yl]carbamoyl}-2,5,8,11-tetraoxatridecan-13-yl)carbamoyl]-
-2-methoxyphenyl} pyrrolidine-2-carboxamide;
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-{4-[(1-{[(2S)-1-[(2S,4R)-4-hydroxy-2-({[4-(4-
-methyl-1,3-thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3,3-dime-
thyl-1-oxobutan-2-yl]carbamoyl}-2,5,8,11-tetraoxatridecan-13-yl)carbamoyl]-
-2-methoxyphenyl} pyrrolidine-2-carboxamide;
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-[4-({2-[2-({[(2S)-1-[(2S,4R)-4-hydroxy-2-({[-
4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3,3--
dimethyl-1-oxobutan-2-yl]carbamoyl}methoxy)ethoxy]ethyl}carbamoyl)-2-metho-
xyphenyl]pyrrolidine-2-carboxamide;
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-[4-({2-[2-({[(2S)-1-[(2S,4R)-4-hydroxy-2-({[-
4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3,3--
dimethyl-1-oxobutan-2-yl]carbamoyl}methoxy)ethoxy]ethyl}carbamoyl)-2-metho-
xyphenyl]pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-{4-[(2-{2-[2-({[(2S)-1-[(2S,4R)-4-hydroxy-2-({[4-(4-methyl-
-1,3-thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1--
oxobutan-2-yl]carbamoyl}methoxy)ethoxy]ethoxy}ethyl)carbamoyl]-2-ethoxyphe-
nyl}pyrrolidine-2-carboxamide;
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-{4-[(2-{2-[2-({[(2S)-1-[(2S,4R)-4-hydroxy-2--
({[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3-
,3-dimethyl-1-oxobutan-2-yl]carbamoyl}methoxy)ethoxy]ethoxy}ethyl)carbamoy-
l]-2-methoxyphenyl}pyrrolidine-2-carboxamide;
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-{4-[(2-{2-[2-({[(2S)-1-[(2S,4R)-4-hydroxy-2--
({[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3-
,3-dimethyl-1-oxobutan-2-yl]carbamoyl}methoxy)ethoxy]ethoxy}ethyl)carbamoy-
l]-2-methoxyphenyl}pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-(3-{[5--
(4-{3-[4-cyano-3-(trifluoromethyl)phenyl]-5,5-dimethyl-4-oxo-2-sulfanylide-
neimidazolidin-1-yl}phenoxy)pentyl]oxy}propyl)-5-(2,2-dimethylpropyl)pyrro-
lidine-2-carboxamide
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-{4-[(3--
{[5-(4-{3-[4-cyano-3-(trifluoromethyl)phenyl]-5,5-dimethyl-4-oxo-2-sulfany-
lideneimidazolidin-1-yl}phenoxy)pentyl]oxy}propyl)carbamoyl]-2-methoxyphen-
yl}-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-(2-methoxy-4-{[2-(2-{[(1,4-trans)-4-({4-[1-benzyl-5-(dimet-
hylamino)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)cyclohexyl]oxy}
ethoxy)ethyl]carbamoyl}phenyl)pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-[2-methoxy-5-({2-[2-(2-{[(1,4-trans)-4-({4-[1-benzyl-5-(di-
methylamino)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)cyclohexyl]oxy}
ethoxy)ethoxy]ethyl}carbamoyl)phenyl]pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-[4-({1--
[4-(3-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]carbamoyl}-5-[e-
thyl(oxan-4-yl)amino]-4-methylphenyl)phenyl]-1,4,7,10-tetraoxadodecan-12-y-
l}carbamoyl)-2-methoxyphenyl]-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxam-
ide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,-
2-dimethylpropyl)-N-[2-methoxy-4-({1-[(1,4-trans)-4-({4-[1-benzyl-5-(dimet-
hylamino)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)cyclohexyl]-1,4,7,10-tetrao-
xadodecan-12-yl}carbamoyl)phenyl]pyrrolidine-2-carboxamide;
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-[2-methoxy-5-({1-[(1,4-trans)-4-({4-[1-benzyl-5-(dimethyla-
mino)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)cyclohexyl]-1,4,7,10,13-pentaox-
apentadecan-15-yl}carbamoyl)phenyl]pyrrolidine-2-carboxamide; and
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-[4-({1--
[4-(3-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]carbamoyl}-5-[e-
thyl(oxan-4-yl)amino]-4-methylphenyl)phenyl]-1,4,7,10,13,16-hexaoxaoctadec-
an-18-yl}carbamoyl)-2-methoxyphenyl]-5-(2,2-dimethylpropyl)pyrrolidine-2-c-
arboxamide, or a pharmaceutically acceptable salt, enantiomer,
stereoisomer, solvate, polymorph or prodrug thereof.
23. A composition comprising an effective amount of the compound of
claim 1.
24. A pharmaceutical composition comprising an effective amount of
a compound of 1 and a pharmaceutically acceptable carrier,
additive, and/or excipient.
25. The pharmaceutical composition of 24, further comprising an
additional bioactive agent, wherein the additional bioactive agent
is an anticancer agent.
26. The composition according to 25, wherein said anticancer agent
is selected from the group consisting of 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 inhibitors, 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, LY 317615, 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)-indolylj-quinolone,
vatalanib, AG-013736, AVE-0005, the acetate salt of [D-Ser(Bu t) 6,
Azgly 10] (pyro-Glu-His-Trp-Ser-Tyr-D-Ser(Bu
t)-Leu-Arg-Pro-Azgly-NH.sub.2 acetate
[C.sub.59H.sub.84N.sub.18Oi.sub.4-(C.sub.2H.sub.4O.sub.2).sub.x
where x=1 to 2.4], 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, gleevac, 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 antagonists,
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.
27. A method for inducing degradation of a target protein in a cell
comprising administering an effective amount of the compound of 1
to the cell.
28. A method for treating a disease state or condition in a patient
wherein dysregulated protein activity is responsible for said
disease state or condition, said method comprising administering an
effective amount of a compound according to claim 1, wherein the
disease state or condition is cancer.
29. The method of 28, wherein the cancer is squamous-cell
carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular
carcinomas, and renal cell carcinomas, cancer of the bladder,
bowel, breast, cervix, colon, esophagus, head, kidney, liver, lung,
neck, ovary, pancreas, prostate, and stomach; leukemias; benign and
malignant lymphomas, particularly Burkitt's lymphoma and
Non-Hodgkin's lymphoma; benign and malignant melanomas;
myeloproliferative diseases; multiple myeloma, sarcomas, including
Ewing's sarcoma, hemangiosarcoma, Kaposi's sarcoma, liposarcoma,
myosarcomas, peripheral neuroepithelioma, synovial sarcoma,
gliomas, astrocytomas, oligodendrogliomas, ependymomas,
gliobastomas, neuroblastomas, ganglioneuromas, gangliogliomas,
medulloblastomas, pineal cell tumors, meningiomas, meningeal
sarcomas, neurofibromas, and Schwannomas; bowel cancer, breast
cancer, prostate cancer, cervical cancer, uterine cancer, lung
cancer, ovarian cancer, testicular cancer, thyroid cancer,
astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer,
liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's
disease, Wilms' tumor or teratocarcinomas, T-lineage Acute
lymphoblastic Leukemia (T-ALL), T-lineage lymphoblastic Lymphoma
(T-LL), Peripheral T-cell lymphoma, Adult T-cell Leukemia, Pre-B
ALL, Pre-B Lymphomas, Large B-cell Lymphoma, Burkitts Lymphoma,
B-cell ALL, Philadelphia chromosome positive ALL and Philadelphia
chromosome positive CML.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
application No. 62/191,193, filed Jul. 10, 2015, the entire
contents of the aforementioned application are hereby incorporated
herein by reference.
INCORPORATION BY REFERENCE
[0002] U.S. patent application Ser. No. 14/686,640, filed Apr. 14,
2015, entitled "Imide-Based Modulators of Proteolysis and
Associated Methods of Use", and PCT Patent Application Serial No.
PCT/US2013/021136, filed Jan. 11, 2013 entitled "Compounds and
Methods for the Enhanced Degradation of Targeted Proteins and Other
Polypeptides by an E3 Ubiquitin Ligase," are incorporated herein by
reference in their entirety. Furthermore, all references cited
herein are incorporated by reference herein in their entirety.
FIELD OF THE INVENTION
[0003] The description provides compounds binding to MDM2,
including bifunctional compounds comprising the same as mentioned,
and associated methods of use. The bifunctional compounds are
useful as modulators of targeted ubiquitination, especially with
respect to a variety of polypeptides and other proteins, which are
degraded and/or otherwise inhibited by bifunctional compounds
according to the present invention.
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 mouse double minute 2 homolog (MDM2)
inhibitors, additional compounds have been reported that target
MDM2 (i.e., human double minute 2 or HDM2) E3 ligases (J. Di, et
al. Current Cancer Drug Targets (2011), 11(8), 987-994).
[0005] Tumor suppressor gene p53 plays an important role in cell
growth arrest and apoptosis in response to DNA damage or stress (A.
Vazquez, et al. Nat. Rev. Drug. Dis. (2008), 7, 979-982), and
inactivation of p53 has been suggested as one of the major pathway
for tumor cell survival (A. J. Levine, et al. Nature (2000), 408,
307-310). In cancer patients, about 50% were found with p53
mutation (M. Hollstein, et al. Science (1991), 233, 49-53), while
patients with wild type p53 were often found p53 down regulation by
MDM2 through the protein-protein interaction of p53 and MDM2 (P.
Chene, et al. Nat. Rev. Cancer (2003), 3, 102-109). Under normal
cell condition without oncogenic stress signal, MDM2 keeps p53 at
low concentration. In response to DNA damage or cellular stress,
p53 level increases, and that also causes increase in MDM2 due to
the feedback loop from p53/MDM2 auto regulatory system. In other
words, p53 regulates MDM2 at the transcription level, and MDM2
regulates p53 at its activity level (A. J. Levine, et al. Genes
Dev. (1993) 7, 1126-1132).
[0006] Several mechanisms can explain p53 down regulation by MDM2.
First, MDM2 binds to N-terminal domain of p53 and blocks expression
of p53-responsive genes (J. Momand, et al. Cell (1992), 69,
1237-1245). Second, MDM2 shuttles p53 from nucleus to cytoplasm to
facilitate proteolytic degradation (J. Roth, et al. EMBO J. (1998),
17, 554-564). Lastly, MDM2 carries intrinsic E3 ligase activity of
conjugating ubiquitin to p53 for degradation through
ubiquitin-dependent 26s proteasome system (UPS) (Y. Haupt, et al.
Nature (1997) 387, 296-299). Therefore, disrupting p53/MDM2 auto
regulation can restore p53 activity and could bring a new approach
in the treatment of cancer. Not surprisingly, since the first
publication of small molecule MDM2 inhibitor Nutlins, multiple
classes of MDM2 antagonists have been reported and several of them
have advanced to the clinic development (B. Zhang, et al. Future
Med. Chem. (2015) 7, 631-645)
[0007] The most studied MDM2 antagonists are imidazolines with
aromatic rings decorated at the three carbons of the ring and NH
group functionalized. One example is RG7112 developed by Roche, in
which two adjacent phenyl rings on imidazoline core are in
cis-conformation (L. T. Vassilev, et al. Science (2004) 57,
1454-1472; B. Vu, et al. ACS Med. Chem. Lett. (2013) 4, 466-469).
The similar cis-bis-aryl substitution pattern is also presented in
Daiichi-Sankyo's MDM2 antagonist DS-5272, although imidazoline core
was replaced with thiazoloimidazoline (M. Miyazaki, et al. Bioorg.
Med. Chem. Lett. (2015) 23, 2360-2367; WO 2014/038606). The earlier
version DS-3032b advanced to clinical testing
(www.clinicaltrials.gov)
[0008] The spiroindolinone compounds MI-219 and MI-888 from
University of Michigan possesses a 5-membered pyrrolidine ring with
two adjacent phenyl ring substituted at the core with cis- and
trans-conformation (S. Wang, et al. PNAS USA (2008) 105,
3933-3938). Further modification in this chemical class resulted in
Sanofi-Aventis' SAR405838 (S. Wang, et al. J. Med. Chem. (2015) 58,
1038-1052; WO 2014/107713).
[0009] Similar to spiroindolinone MDM2 antagonist, non-spiro
molecules with pyrrolidine core decorated by adjacent
trans-bis-aryl rings were reported by Roche, and RG7388 from this
chemical series became Roche's second MDM2 inhibitor in the clinic
(Q. Ding, et al. J. Med. Chem. (2013) 56, 5979-5983)
[0010] Piperidinone and morpholinone cores with adjacent trans-aryl
substitution on the core are another chemical class of MDM2
inhibitors reported by Amgen. These compounds are structurally
different from imidazoline or spiro-indolinone or pyrrolidine
chemical class. AMG-232 with a piperidinone core advanced to the
clinic (D. Sun, et al. J. Med. Chem. (2014) 57, 1454-1472). AM-7209
is a more potent molecule from Amgen reported recently (Y. Rew, et
al. J. Med. Chem. (2014) 57, 10499-10511). A diversity of
structures with 6-membered cores were reported by Amgen (WO
2014/151863, WO2014/134201, US 2014/235629, US 2014/0243372).
[0011] Pyrrolopyrimidine- and imidazolopyridine-derived carboxylic
acid and acid mimetic oxadiazolone analogs are potent HDM2
inhibitors reported by Merck (WO 2014/100065; WO 2014/100071).
MK-8242, also known as SCH90042, has been tested in the clinic
(www.clinicaltrials.gov).
[0012] Novartis reported pyrrolopyrrolidinone chemical class where
three aryl groups were attached to pyrrolopyrrolidinone core (WO
2013/175417). CMG097, also known as NVP-CMG-097 in the clinic, is a
small molecule MDM2 inhibitor derived from 1,2-bis-aryl-substituted
dihydro-isoquinolinone chemical class (WO 2014/020502).
[0013] All small molecule MDM2 inhibitors mentioned above showed
potent activity in inhibiting p53 and MDM2 interaction, which
consequently stabilizes p53. However, due to the feedback loop,
antagonism mode also resulted in MDM2 up-regulation at the
transcription level as shown in the literature. As such, the
potential exists that degrading MDM2 could overcome issues
associated with MDM2 up-regulation. Also, because MDM2 functions as
E3 ligase, recruiting MDM2 to a disease causing protein and
effectuating its ubiquitination and degradation is an approach of
high interest for drug discovery.
[0014] An ongoing need exists in the art for effective treatments
for disease, especially hyperplasias and cancers. However,
non-specific effects, and the inability to target and modulate
certain classes of proteins altogether, such as transcription
factors, remain as obstacles to the development of effective
anti-cancer agents. As such, small molecule therapeutic agents that
leverage or potentiate MDM2 substrate specificity and, at the same
time, are "tunable" such that a wide range of protein classes can
be targeted and modulated with specificity would be very useful as
a therapeutic.
SUMMARY
[0015] The present disclosure describes bifunctional compounds
which function to recruit endogenous proteins to an E3 ubiquitin
ligase for degradation, and methods of using the same. In
particular, the present disclosure provides bifunctional or
proteolysis targeting chimeric (PROTAC) compounds, which find
utility as modulators of targeted ubiquitination of a variety of
polypeptides and other proteins, which are then degraded and/or
otherwise inhibited by the bifunctional compounds as described
herein. An advantage of the 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, e.g., multiple myeloma.
[0016] As such, in one aspect the disclosure provides MDM2
ligand-based PROTAC compounds of Formula (A):
##STR00001##
[0017] Formula (A) represents bifunctional or PROTAC compounds,
which comprise an E3 ubiquitin ligase binding moiety (i.e., a
ligand for an E3 ubquitin ligase or "ULM" group), coupled via
linker (L) to a moiety that binds a target protein (i.e., a
protein/polypeptide targeting ligand or "PTM" group) such that the
target protein/polypeptide is placed in proximity to the ubiquitin
ligase to effect degradation (and inhibition) of that protein. In a
preferred embodiment, the ULM is a moiety that binds MDM2 E3
ubiquitin ligase (i.e., "MLM").
[0018] In Formula (A), the respective positions of the PTM and MLM
moieties 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.
[0019] In certain preferred embodiments, PTM is a protein target
moiety. As such, PTM binds to a specific protein which is set to be
ubiquitinated or degraded.
[0020] In certain embodiments, "L" is the linker that connects PTM
and MLM. In certain embodiments, L is a bond (i.e., absent). In
certain additional embodiments, L is a chemical linker as described
herein. In certain preferred 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.
[0021] In certain additional embodiments, the MLM of the
bifunctional compound with a formula (A) 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.
[0022] In additional embodiments, the MLM comprises the core
structures mentioned above with adjacent bis-aryl substitutions
positioned as cis- or trans-configurations.
[0023] 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.
[0024] In certain embodiments, the compounds as described herein
comprise multiple MLMs, multiple PTMs, multiple chemical linkers or
a combination thereof.
[0025] In certain embodiments, PTMs can be, but not limited to,
small molecules binding to kinases, enzymes, transporters, nuclear
hormone receptors, non-nuclear hormone receptors, G-protein coupled
receptors (GPCRs), transcription factors, and epigenetic
targets.
[0026] In certain embodiments, the epigenetic targets can be
bromodomain and extra terminal domain (BET) family proteins, such
as, e.g., BRD1, -2, -3, or -4.
[0027] In certain embodiments, the nuclear hormone receptors can
be, but not limited to, androgen receptor (AR) and estrogen
receptor (ER).
[0028] In another aspect, the description provides bifunctional
molecules as shows in Formula (B), wherein PTM comprises an MDM2
binding moiety (MBM) coupled via a linker (L) to ULM
(ubiquitination ligase binding moiety), which comprises a moiety
that binds an E3 ubiquitin ligase, e.g., Von Hippel-Lindau E3
ubiquitin ligase (VHM), Cereblon (CLM) or MDM2 (MLM).
##STR00002##
[0029] In certain embodiments, "L" is the linker that connects PTM
and MLM. In certain embodiments, L is a bond (i.e., absent). In
certain additional embodiments, L is a chemical linker as described
herein. In certain preferred 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.
[0030] In an embodiment, VLM can be hydroxyproline or a derivative
thereof. Other contemplated VLMs are described in U.S. Patent
Application Pub. No. 2014/03022523A1, and 2015/0291562A1, which are
incorporated herein in their entirety.
[0031] In certain embodiments, MBM 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.
[0032] In additional embodiments, the MBM comprises the core
structures mentioned above with adjacent bis-aryl substitutions
positioned as cis- or trans-configurations.
[0033] In still additional embodiments, the MBM 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.
[0034] 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.
[0035] 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
[0036] 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.
[0037] In yet another aspect, the description provides a method of
ubiquitinating/degrading a target protein in a cell. In certain
embodiments, the method comprises administering to a subject or
contacting a subject, e.g., a patient or a cell, with a
bifunctional compound as described herein, wherein the bifunctional
compound effectuates degradation of the target protein. 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 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.
[0038] In still another aspect, the description provides methods
for treating or emeliorating a disease, disorder or symptom thereof
in a subject or a patient, e.g., an animal such as a mammal or 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.
[0039] 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
invention.
[0040] 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 invention 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
invention may be utilized in numerous combinations, all of which
are expressly contemplated by the present description. These
additional advantages objects and embodiments are expressly
included within the scope of the present invention. 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
[0041] The accompanying drawings, which are incorporated into and
form a part of the specification, illustrate several embodiments of
the present invention 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:
[0042] FIG. 1: c-Myc suppression in 22rv1 cells by chimeric
molecules, where BRD4 ligand is connected through linkers to MDM2
ligands using partial structural motif in RG7388. Chimeric
molecules with inactive MDM2 ligand (enantiomer of the active
counterpart) demonstrated no c-Myc suppression across a range of
concentrations, while chimeric molecules with active MDM2 ligand
showed dose dependent c-Myc suppression, suggesting BRD4
degradation mediated by MDM2 E3 ligase ubiquitination mechanism, as
c-Myc is directly regulated by the level of BRD4. Chimeric
molecules with MDM2 ligand as a racemate displayed similar c-Myc
suppression as observed in those containing active MDM2 ligand.
[0043] FIG. 2: Western blot of HCT116 cells treated with chimeric
molecules, where BRD4 ligand is connected through linkers to MDM2
ligands using partial structural motif in RG7388. Chimeric
molecules with inactive MDM2 ligand (A-1891, A-1894) demonstrated
no p53 level increase and no MDM2 up-regulation, while chimeric
molecules with active MDM2 ligand (A-1864, A1892 and A-1893, A-1877
carried a racemic MDM2 binding ligand) showed dose dependent p53
level increase and up-regulation of MDM2, suggesting chimeric
molecules with BRD4 binding fragment and MDM2 binding fragment
connected through a linker can function as small molecule MDM2
antagonist in stabilizing p53. The less significant MDM2 up
regulation and p53 level increase is due to the chimeric molecule
action mechanism of not only binding to MDM2 to block p53-MDM2
interaction but also degrading MDM2. Therefore, the net MDM2
up-regulation is significantly less, which also translated to p53
level due to MDM2-p53 feedback loop.
[0044] FIG. 3: Western blot of HCT116 cells treated with chimeric
molecules, where MDM2 ligand (using partial structural motif of
RG7388) is connected through linkers to VHL ligand. Chimeric
molecules with inactive MDM2 ligand (A-1897, A1908, and A-1911)
demonstrated no p53 level increase and no MDM2 up-regulation, while
chimeric molecules with active MDM2 ligand (A-1896, A-1907, and
A-1910, with A-1877, A-1895, and A-1909 carrying a racemic MDM2
binding ligand) showed dose dependent p53 level increase.
[0045] FIG. 4: Inhibition of cell proliferation in HCT116 and 22rv1
cells by chimeric molecules containing MDM2 binding motif. In
p53.sup.WT HCT-116 colon cancer cell lines, MDM2-recruiting BRD-4
PROTAC with active MDM2 binding moiety (A-1893) caused very potent
growth inhibition in comparison with the MDM2-recruiting BRD-4
PROTAC with inactive MDM2 binding moiety (A-1894). In this cell
growth assay, BRD4-Cereblon PROTAC A-825, MDM2 antagonist RG7388
(A-1850), the racemate of RG7388 (A-1851) and JQ1 were included as
a direct comparison.
[0046] FIG. 5: Time course of BRD4 degradation caused by BRD4-MDM2
chimeric compound (A-1893) in human colon cancer cell line
HCT116.
[0047] FIG. 6: Time course of BRD4 degradation caused by BRD4-MDM2
chimeric compound (A-1893) in human lung cancer cell line A549.
DETAILED DESCRIPTION
[0048] 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.
[0049] The present disclosure describes bifunctional compounds
which function to recruit endogenous proteins to an E3 ubiquitin
ligase for degradation, and methods of using the same. In
particular, the present disclosure provides bifunctional or
proteolysis targeting chimeric (PROTAC) compounds, which find
utility as modulators of targeted ubiquitination of a variety of
polypeptides and other proteins, which are then degraded and/or
otherwise inhibited by the bifunctional compounds as described
herein. An advantage of the 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.
[0050] As such, presently described are compositions and methods
that relate to the surprising and unexpected discovery that an E3
ubiquitin ligase protein, e.g., MDM2, ubiquitinates a target
protein once it and the target protein are placed in proximity by a
bifunctional or chimeric construct (e.g., a PROTAC) that binds the
E3 ubiquitin ligase protein and the target protein. Accordingly,
the present invention provides such compounds and compositions
comprising an E3 ubiquintin 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. The present
invention also provides a library of compositions and the use
thereof.
[0051] In particular, the present application is directed to
compounds which contain 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 MDM2, and a moiety that is capable of binding to a 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.
[0052] 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.
[0053] 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.
[0054] 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.
[0055] 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.
[0056] 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.
[0057] 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."
[0058] 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.
[0059] 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
non-limiting 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.
[0060] 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.
[0061] 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 co-administered 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.
[0062] 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 steroisomers (diastereomers) thereof, as
well as pharmaceutically acceptable salts and derivatives
(including prodrug forms) thereof where applicable, in context.
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 within
the context of the compound shown.
[0063] The term "compound" or "chemical compound" as used herein
can include organometallic compounds, organic compounds, metals,
transitional metal complexes, and small molecules. In certain
preferred embodiments, polynucleotides are excluded from the
definition of compounds. In other preferred embodiments,
polynucleotides and peptides are excluded from the definition of
compounds. In a particularly preferred embodiment, the term
compounds refers to small molecules (e.g., preferably, non-peptidic
and non-oligomeric) and excludes peptides, polynucleotides,
transition metal complexes, metals, and organometallic
compounds.
[0064] As used herein, the term "small molecule" refers to a
non-peptidic, non-oligomeric organic compound either synthesized in
the laboratory or found in nature. Small molecules, as used herein,
can refer to compounds that are "natural product-like", however,
the term "small molecule" is not limited to "natural product-like"
compounds. Rather, a small molecule is typically characterized in
that it contains several carbon-carbon bonds, and has a molecular
weight of less than 2000 g/mol, preferably less than 1500 g/mol,
although this characterization is not intended to be limiting for
the purposes of the present application. In certain other preferred
embodiments, synthetic small molecules are utilized.
[0065] 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, MDM2 is 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. The most common lysine is Lys48 on the
ubiquitin chain. This is the lysine used to make polyubiquitin,
which is recognized by the proteasome.
[0066] 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 invention
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
invention, the term patient refers to a human patient unless
otherwise stated or implied from the context of the use of the
term.
[0067] 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.
DETAILED DESCRIPTION
[0068] In one aspect, the description provides compounds comprising
an MDM2 E3 ubiquitin ligase binding moiety (MLM) connected to a
linker (L), as shown below, wherein MLM is a ligand for MDM2 or
HDM2, and L is a bond or a chemical linker group.
##STR00003##
[0069] The number and/or relative positions of the moieties in the
compounds illustrated herein are provided by way of example only.
As would be understood by the skilled artisan, compounds as
described herein can be synthesized with any desired number and/or
relative position of the respective functional moieties.
[0070] In another aspect, the description provides compounds
comprising an MDM2 E3 ubiquitin ligase binding moiety (MLM) coupled
via a linker (L) to a protein targeting moiety (PTM), wherein L is
a bond or a chemical linker group. These compounds are described
herein as "MDM2 PROTAC compounds" (MDM2-mediated proteolysis
targeting chimerics) and are presented as Formula (A):
##STR00004##
[0071] In Formula (A), the respective positions of the PTM and MLM
moieties 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.
[0072] In Formula (A), PTM is a protein/polypeptide targeting
moiety, L is a linker, and MLM is a MDM2 E3 ubiquitin ligase
binding moiety.
[0073] In another aspect, the description provides bifunctional
molecules as shows in Formula (B), wherein PTM comprises an MDM2
binding moiety (MBM) coupled via a linker (L) to ULM
(ubiquitination ligase binding moiety), which comprises a moiety
that binds an E3 ubiquitin ligase, e.g., Von Hippel Lindau E3
Ligase (VHM), Cereblon (CLM) or MDM2 (MLM).
##STR00005##
[0074] The terms ULM is used inclusively unless the context
indicates otherwise to indicate an E3 ubiquitin ligase binding
moiety, including those that bind MDM2 (i.e., MLMs). Further, the
term MLM is inclusive of all possible MDM2 E3 ubiquitin ligase
binding moieties.
[0075] In certain embodiments, the E3 ubiquitin ligase is MDM2. As
such, the ULM is an MLM that binds to MDM2.
[0076] In certain preferred embodiments, PTM is a protein target
moiety. As such, PTM binds to a specific protein which is set to be
ubiquitinated or degraded.
[0077] In certain preferred embodiments, "L" is a linker, e.g., a
bond (i.e., absent) or a chemical linker that connects PTM and
MLM.
[0078] In certain additional embodiments, the MLM of the
bifunctional compound as depicted in Formula (A) or (B) 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.
[0079] In additional embodiments, the MLM comprises the core
structures mentioned above with adjacent bis-aryl substitutions
positioned as cis- or trans-configurations.
[0080] 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.
[0081] In certain embodiments, the linker "L" is a connector with a
linear non-hydrogen atom number in the range of 1 to 20. In
additional embodiments, the connector "L" comprises a functional
group, e.g., an ether, amide, alkane, alkene, alkyne, ketone,
hydroxyl, carboxylic acid, thioether, sulfoxide, and sulfone. The
linker can also comprise aromatic, heteroaromatic, cyclic, bycyclic
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.
[0082] In certain embodiments, the compounds as described herein
comprise multiple MLMs, multiple PTMs, multiple chemical linkers or
a combination thereof.
[0083] In certain embodiments, PTMs can be, but not limited to,
small molecules binding to kinases, enzymes, transporters, nuclear
hormone receptors, non-nuclear hormone receptors, G-protein coupled
receptors (GPCRs), transcription factors, and epigenetic
targets.
[0084] In certain embodiments, PTM is a small molecule binding to
epigenetic targets, and the epigenetic targets can be BRDs, such as
BRD4.
[0085] In certain embodiments, PTM is a small molecule binding to
nuclear hormone receptors, and the nuclear hormone receptor can be,
but not limited to, androgen receptor (AR) and estrogen receptor
(ER).
[0086] 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.
[0087] 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).
##STR00006##
wherein above Formula (A-1) through Formula (A-8), X is selected
from the group consisting of carbon, oxygen, sulfur, sulfoxide,
sulfone, and N--R.sup.a; [0088] R.sup.a is independently H or an
alkyl group with carbon number 1 to 6; Y and Z are independently
carbon or nitrogen; A, A' and A'' are independently selected from
C, N, O or S, can also be one or two atoms forming a fused bycyclic
ring, or a 6,5- and 5,5-fused aromatic bicyclic group; R.sub.1,
R.sub.2 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: [0089]
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 are
independently selected from the group consisting of H, methyl and
C1 to C6 alkyl; R.sub.5 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: [0090]
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 is H or --C(.dbd.O)R.sup.b, wherein
[0091] R.sup.b 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 [0092] R.sup.c 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 [0093] R.sup.d 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 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.sub.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: [0094] R.sup.e is
an alkylene with 1 to 6 carbons, or a bond; [0095] R.sup.f is a
substituted 4- to 7-membered heterocycle; [0096] R.sup.g is
selected from the group consisting of aryl, hetero aryl,
substituted aryl or heteroaryl, and 4- to 7-membered heterocycle;
R.sub.9 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 consistin of halogen, alkene, alkyne, alkyl,
unsubstituted or substituted with Cl or F; R.sub.10 is selected
from the group consistin 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 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: [0097] 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
[0098] R.sup.h and R.sup.i 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 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 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.sub.15 is CN;
[0099] R.sub.16 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(allkyl), --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.dbd.O)-- 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 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, hydrxylated 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, C1-6 alkyl with hydrogen replaced by a cyano group, 5
and 6 membered aryl or heteroaryl, aklyl 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 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 is
selected from the group consisting of aryl, heteroaryl, bicyclic
heteroaryl, and these aryl or hetroaryl 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 are independently selected
from C1-6 alkyl, C1-6 cycloalkyl, C1-6 alkoxy, hydoxylated 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 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 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, hydoxylated
C1-6 alkyl, cycloalkyl, fluorine-substituted C1-6 alkyl, CN,
alkoxy, alkyl sulfone, amide and sulfonamide; R.sub.24 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, hydoxylated
alkyl, cyano-substituted alkyl, cycloalyl and substituted
cycloalkyl; R.sub.25 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 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 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 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'' is selected from the group consisting
of alkyl, aryl substituted alkyl, alkoxy substituted alkyl,
cycloalkyl, aryl-substituted cycloalkyl, and alkoxy substituted
cycloalkyl.
[0100] In certain embodiments, the heterocycles in R.sup.f and
R.sup.g are substituted pyrrolidine, substituted piperidine,
substituted piperizine.
[0101] Unless the context indicates otherwise, the following terms
can mean:
[0102] The term "independently" is used herein to indicate that the
variable, which is independently applied, varies independently from
application to application.
[0103] 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 (Br, Cl, F, or I).
[0104] The term "lower alkyl" refers to methyl, ethyl or propyl
[0105] The term "lower alkoxy" refers to methoxy, ethoxy or
propoxy.
[0106] 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.
[0107] 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.
[0108] 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,
.beta.-alanine, arginine, asparagine, aspartic acid, cysteine,
cystine, glutamic acid, glutamine, glycine, phenylalanine,
histidine, isoleucine, lysine, leucine, methionine, proline,
serine, threonine, valine, tryptophan or tyrosine.
[0109] 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 C.sub.0, H stands in place of
carbon.
[0110] 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 substituents, preferably up
to three substituents, often 1 or 2 substituents on a moiety in a
compound according to the present invention 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 (CN), 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 invention 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 --(CH.sub.2).sub.m-- or --(CH.sub.2).sub.m-- groups or other
chains such as ethylene glycol chains, as identified above, may be
substituted anywhere on the chain. Preferred substituents 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 side chain 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 substituents, 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 invention, a moiety in a molecule may be optionally
substituted with up to five substituents, preferably up to three
substituents. Most often, in the present invention moieties which
are substituted are substituted with one or two substituents.
[0111] 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
[0112] 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)OH, --(CH.sub.2)--SH, --(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)OH,
--(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)C(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.
[0113] 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 invention 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, indoline, azaindoline,
benzofuran, 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, indoline, azaindoline, 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.
[0114] 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, CO 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 methylsubstitutedpyridine 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.
[0115] "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.
[0116] 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.
##STR00007##
wherein [0117] S.sup.c is CHR.sup.SS, NR.sup.URE, or O; [0118]
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); [0119] 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); [0120]
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 [0121] 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).
[0122] 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.
[0123] 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, pyrrolidinyl, pyrrolinyl, pyrrolyl, quinolinyl,
tetrahydrofuranyl, tetrahydropyranyl, tetrahydroquinoline,
thiazolidinyl, thiazolyl, thienyl, tetrahydrothiophene, oxane,
oxetanyl, oxathiolanyl, thiane among others.
[0124] 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, --SO.sub.2-alkyl, --SO.sub.2-substituted alkyl,
--SO.sub.2-aryl, oxo (.dbd.O), and --SO.sub.2-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).
[0125] 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.
[0126] "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.
[0127] 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.
[0128] Using MBM or 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.
[0129] In certain embodiments, the description provides a
bifunctional molecule comprising a structure selected from the
group consisting of:
##STR00008##
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'' are
as defined herein.
[0130] 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:
##STR00009##
wherein R1' and R2' 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' 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' 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'
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.
[0131] In certain embodiments, the linker connection position is at
least one of R4' or R6' or both.
[0132] In certain embodiments, R6' is independently selected from
the group consisting of H,
##STR00010## ##STR00011##
wherein "*" indicates the point of attachment of the linker.
[0133] In certain embodiments, the linker is attached to at least
one of R1', R2', R3', R4', R5', R6', or a combination thereof.
[0134] 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:
##STR00012## ##STR00013##
wherein: R7' is a member selected from the group consisting of
halogen, mono-, and di- or tri-substituted halogen; R8' 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' is
selected from the group consistin 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 is
selected from the group consistin of H, --OCH.sub.3,
--OCH.sub.2CH.sub.3, and halogen; R10' and R11' 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.in(CH.sub.2).sub.n--COOH,
(CH.sub.2CH.sub.2O).sub.in(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), --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)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).sub.2, oxo, carboxy, clcloalkyl and heteroaryl; m, n, and
p are independently 0 to 6; R12' is selected from the group
consisting of --O-(alkyl), --O-(alkyl)-akoxy, --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'' is selected from the
group consisting of alkyl, aryl substituted alkyl, aloxy
substituted alkyl, cycloalkyl, ary-substituted cycloalkyl, and
alkoxy substituted cycloalkyl.
[0135] In any of the aspects or embodiments described herein, the
alkyl, alkoxy or the like can be a lower alkyl or lower alkoxy.
[0136] In certain embodiments, the linker connection position is at
least one of Z, R8', R9', R10', R11'', R12'', or R1''.
[0137] 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 MBM or
MLM with formula A-2, A-3, A-5, A-6, A-7 and A-8, wherein the
solvent exposed area in the MBM or MLM can be connected to linker
"L" which will be attached to target protein ligand "PTM", to
construct PROTACs.
[0138] Exemplary Linkers
[0139] In certain embodiments, the compounds as described herein
can be chemically linked or coupled via a chemical linker (L). In
certain embodiments, the linker group L is a group comprising one
or more covalently connected structural units of B (e.g., --B.sub.1
. . . B.sub.q--), wherein B.sub.1 is a group coupled to at least
one of a MBM, a PTM, or a combination thereof. In certain
embodiments, B.sub.1 links an MBM, a PTM, or a combination thereof.
In certain embodiments B.sub.1 links an MBM, a PTM or a combination
thereof directly to another MBM, PTM, or combination thereof. In
other embodiments, B.sub.1 links a MBM, a PTM, or a combination
thereof indirectly to another MBM, PTM, or combination thereof
through B.sub.q.
[0140] In certain embodiments, B.sub.1 to B.sub.q are, each
independently, 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.3-11heteocyclyl optionally
substituted with 0-6 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, can be
linked to other B groups to form cycloalkyl and/or heterocyclyl
moeity which can be further substituted with 0-4 R.sup.1-5 groups;
wherein [0141] 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.1-8 alkyl, 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.
[0142] In certain embodiments, q is an integer greater than or
equal to 0. In certain embodiments, q is an integer greater than or
equal to 1.
[0143] In certain embodiments, e.g., where q is greater than 2,
B.sub.q is a group which is connected to MBM, and B.sub.1 and
B.sub.q are connected via structural units of B (number of such
structural units of B: q-2).
[0144] In certain embodiments, e.g., where q is 2, B.sub.q is a
group which is connected to B.sub.1 and to a MBM.
[0145] In certain embodiments, e.g., where q is 1, the structure of
the linker group L is --B.sub.1--, and B.sub.1 is a group which is
connected to a MBM moiety and a PTM moiety.
[0146] In additional embodiments, q is an integer from 1 to 100, 1
to 90, 1 to 80, 1 to 70, 1 to 60, 1 to 50, 1 to 40, 1 to 30, 1 to
20, or 1 to 10.
[0147] In certain embodiments, the linker (L) is selected from the
group consisting of:
[0148] --(CH.sub.2).sub.n-(lower alkyl)-, --(CH.sub.2).sub.n-(lower
alkoxyl)-, --(CH.sub.2).sub.n-(lower alkoxyl)-OCH.sub.2--C(O)--,
--(CH.sub.2).sub.n-(lower alkoxyl)-(lower alkyl)-OCH.sub.2--C(O)--,
--(CH.sub.2).sub.n-(cycloalkyl)-(lower alkyl)-OCH.sub.2--C(O)--,
--(CH.sub.2).sub.n-(hetero cycloalkyl)-,
--(CH.sub.2CH.sub.2O).sub.n-(lower alkyl)-O--CH.sub.2--C(O)--,
--(CH.sub.2CH.sub.2O).sub.n-(hetero
cycloalkyl)-O--CH.sub.2--C(O)--,
--(CH.sub.2CH.sub.2O).sub.n-Aryl-O--CH.sub.2--C(O)--,
--(CH.sub.2CH.sub.2O).sub.n-(hetero aryl)-O--CH.sub.2--C(O)--,
--(CH.sub.2CH.sub.2O).sub.n-(cyclo alkyl)-O-(hetero
aryl)-O--CH.sub.2--C(O)--, --(CH.sub.2CH.sub.2O).sub.n-(cyclo
alkyl)-O-Aryl-O--CH.sub.2--C(O)--,
--(CH.sub.2CH.sub.2O).sub.n-(lower
alkyl)-NH-Aryl-0-CH.sub.2--C(O)--,
--(CH.sub.2CH.sub.2O).sub.n-(lower alkyl)-O-Aryl-C(O)--,
--(CH.sub.2CH.sub.2O).sub.n-cycloalkyl-O-Aryl-C(O)--,
--(CH.sub.2CH.sub.2O).sub.n-cycloalkyl-O-(hetero aryl)1-C(O)--,
where n can be 0 to 10
##STR00014## ##STR00015## ##STR00016##
[0149] 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.
[0150] 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.
[0151] Although the MLM (or 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 invention, the linker is independently
covalently bonded to the MLM 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 MLM group and PTM group to provide maximum binding
of the MLM 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 MLM and/or PTM groups.
[0152] In certain embodiments, "L" can be linear chains with linear
atoms from 4 to 24, the carbon atom in the linear chain can be
substituted with oxygen, nitrogen, amide, fluorinated carbon, etc.,
such as the following:
##STR00017## ##STR00018## ##STR00019##
[0153] In certain embodiments, "L" can be nonlinear chains, and can
be aliphatic or aromatic or heteroaromatic cyclic moieties, some
examples of "L" include but not be limited to the following:
##STR00020## ##STR00021##
wherein: [0154] "X" in above structures can be linear chain with
atoms ranging from 2 to 14, and the mentioned chain can contain
heteroatoms such as oxygen; and [0155] "Y" in above structures can
be O, N, S(O).sub.n (n=0, 1, 2).
[0156] Exemplary PTMs
[0157] In preferred aspects of the invention, the PTM group is a
group, which binds to target proteins. 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 invention.
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 invention. Preferably, the target protein is a
eukaryotic protein. In certain aspects, the protein binding moiety
is a haloalkane (preferably a C.sub.1-C.sub.10 alkyl group which is
substituted with at least one halo group, preferably a halo group
at the distal end of the alkyl group (i.e., away from the linker or
CLM group), which may covalently bind to a dehalogenase enzyme in a
patient or subject or in a diagnostic assay.
[0158] PTM groups according to the present invention include, for
example, include any moiety which binds to a protein specifically
(binds to a target protein) and includes the following non-limiting
examples of small molecule target protein moieties: Hsp90
inhibitors, kinase inhibitors, compounds targeting Human BET
Bromodomain-containing proteins, HDAC inhibitors, human lysine
methyltransferase inhibitors, angiogenesis inhibitors, nuclear
hormone receptor compounds, immunosuppressive compounds, and
compounds targeting the aryl hydrocarbon receptor (AHR), among
numerous others. The compositions described below exemplify some of
the members of these nine types of small molecule target protein
binding moieties. Such small molecule target protein binding
moieties also include pharmaceutically acceptable salts,
enantiomers, solvates and polymorphs of these compositions, as well
as other small molecules that may target a protein of interest.
These binding moieties are linked to the ubiquitin ligase binding
moiety preferably through a linker in order to present a target
protein (to which the protein target moiety is bound) in proximity
to the ubiquitin ligase for ubiquitination and degradation.
[0159] Any protein, which can bind to a protein target moiety or
PTM group and acted on or degraded by an ubiquitin ligase is a
target protein according to the present invention. In general,
target proteins may include, for example, structural proteins,
receptors, enzymes, cell surface proteins, proteins pertinent to
the integrated function of a cell, including proteins involved in
catalytic activity, aromatase activity, motor activity, helicase
activity, metabolic processes (anabolism and catrabolism),
antioxidant activity, proteolysis, biosynthesis, proteins with
kinase activity, oxidoreductase activity, transferase activity,
hydrolase activity, lyase activity, isomerase activity, ligase
activity, enzyme regulator activity, signal transducer activity,
structural molecule activity, binding activity (protein, lipid
carbohydrate), receptor activity, cell motility, membrane fusion,
cell communication, regulation of biological processes,
development, cell differentiation, response to stimulus, behavioral
proteins, cell adhesion proteins, proteins involved in cell death,
proteins involved in transport (including protein transporter
activity, nuclear transport, ion transporter activity, channel
transporter activity, carrier activity, permease activity,
secretion activity, electron transporter activity, pathogenesis,
chaperone regulator activity, nucleic acid binding activity,
transcription regulator activity, extracellular organization and
biogenesis activity, translation regulator activity. Proteins of
interest can include proteins from eurkaryotes and prokaryotes
including humans as targets for drug therapy, other animals,
including domesticated animals, microbials for the determination of
targets for antibiotics and other antimicrobials and plants, and
even viruses, among numerous others.
[0160] In still other embodiments, the PTM group is a haloalkyl
group, wherein said alkyl group generally ranges in size from about
1 or 2 carbons to about 12 carbons in length, often about 2 to 10
carbons in length, often about 3 carbons to about 8 carbons in
length, more often about 4 carbons to about 6 carbons in length.
The haloalkyl groups are generally linear alkyl groups (although
branched-chain alkyl groups may also be used) and are end-capped
with at least one halogen group, preferably a single halogen group,
often a single chloride group. Haloalkyl PT, groups for use in the
present invention are preferably represented by the chemical
structure --(CH.sub.2).sub.v-Halo where v is any integer from 2 to
about 12, often about 3 to about 8, more often about 4 to about 6.
Halo may be any halogen, but is preferably Cl or Br, more often
Cl.
[0161] In another embodiment, the present invention provides a
library of compounds. The library comprises more than one compound
wherein each composition has a formula of A-B, wherein A is a
ubiquitin pathway protein binding moiety (preferably, an E3
ubiquitin ligase moiety as otherwise disclosed herein) and B is a
protein binding member of a molecular library, wherein A is coupled
(preferably, through a linker moiety) to B, and wherein the
ubiquitin pathway protein binding moiety recognizes an ubiquitin
pathway protein, in particular, an E3 ubiquitin ligase, such as
cereblon. In a particular embodiment, the library contains a
specific cereblon E3 ubiquitin ligase binding moiety bound to
random target protein binding elements (e.g., a chemical compound
library). As such, the target protein is not determined in advance
and the method can be used to determine the activity of a putative
protein binding element and its pharmacological value as a target
upon degradation by ubiquitin ligase.
[0162] The present invention may be used to treat a number of
disease states and/or conditions, including any disease state
and/or condition in which proteins are dysregulated and where a
patient would benefit from the degradation of proteins.
[0163] 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 multiple myeloma.
[0164] In alternative aspects, the present invention 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 invention may be used to treat a large
number of disease states or conditions including cancer, 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.
[0165] 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
invention.
[0166] The term "target protein" is used to describe a protein or
polypeptide, which is a target for binding to a compound according
to the present invention and degradation by ubiquitin ligase
hereunder. Such small molecule target protein binding moieties also
include pharmaceutically acceptable salts, enantiomers, solvates
and polymorphs of these compositions, as well as other small
molecules that may target a protein of interest. These binding
moieties are linked to CLM or ULM groups through linker groups
L.
[0167] 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.
Target proteins include proteins and peptides having any biological
function or activity including structural, regulatory, hormonal,
enzymatic, genetic, immunological, contractile, storage,
transportation, and signal transduction. In certain embodiments,
the target proteins include structural proteins, receptors,
enzymes, cell surface proteins, proteins pertinent to the
integrated function of a cell, including proteins involved in
catalytic activity, aromatase activity, motor activity, helicase
activity, metabolic processes (anabolism and catrabolism),
antioxidant activity, proteolysis, biosynthesis, proteins with
kinase activity, oxidoreductase activity, transferase activity,
hydrolase activity, lyase activity, isomerase activity, ligase
activity, enzyme regulator activity, signal transducer activity,
structural molecule activity, binding activity (protein, lipid
carbohydrate), receptor activity, cell motility, membrane fusion,
cell communication, regulation of biological processes,
development, cell differentiation, response to stimulus, behavioral
proteins, cell adhesion proteins, proteins involved in cell death,
proteins involved in transport (including protein transporter
activity, nuclear transport, ion transporter activity, channel
transporter activity, carrier activity, permease activity,
secretion activity, electron transporter activity, pathogenesis,
chaperone regulator activity, nucleic acid binding activity,
transcription regulator activity, extracellular organization and
biogenesis activity, translation regulator activity. Proteins of
interest can include proteins from eurkaryotes and prokaryotes,
including microbes, viruses, fungi and parasites, including humans,
microbes, viruses, fungi and parasites, among numerous others, as
targets for drug therapy, other animals, including domesticated
animals, microbials for the determination of targets for
antibiotics and other antimicrobials and plants, and even viruses,
among numerous others.
[0168] The term "protein target moiety" or PTM is used to describe
a small molecule which binds to a target protein or other protein
or polypeptide of interest and places/presents that protein or
polypeptide in proximity to an ubiquitin ligase such that
degradation of the protein or polypeptide by ubiquitin ligase may
occur. Non-limiting examples of small molecule target protein
binding moieties include Hsp90 inhibitors, kinase inhibitors, MDM2
inhibitors, compounds targeting Human BET Bromodomain-containing
proteins, HDAC inhibitors, human lysine methyltransferase
inhibitors, angiogenesis inhibitors, immunosuppressive compounds,
and compounds targeting the aryl hydrocarbon receptor (AHR), among
numerous others. The compositions described below exemplify some of
the members of these nine types of small molecule target
protein.
[0169] Exemplary protein target moieties according to the present
disclosure include, haloalkane halogenase inhibitors, Hsp90
inhibitors, kinase inhibitors, MDM2 inhibitors, compounds targeting
Human BET Bromodomain-containing proteins, HDAC inhibitors, human
lysine methyltransferase inhibitors, angiogenesis inhibitors,
immunosuppressive compounds, and compounds targeting the aryl
hydrocarbon receptor (AHR).
[0170] Additional exemplary protein targets to which a PTM may bind
and may be incorporated into compounds as described herein include,
Adrenomedullin (AM), Angiopoietin (Ang), Autocrine motility factor,
Bone morphogenetic proteins (BMPs), Ciliary neurotrophic factor
family Ciliary neurotrophic factor (CNTF), Leukemia inhibitory
factor (LIF), Interleukin-(IL-6), Colony-stimulating factors
Macrophage colony-stimulating factor (m-CSF), Granulocyte
colony-stimulating factor (G-CSF), Granulocyte macrophage
colony-stimulating factor (GM-CSF), Epidermal growth factor (EGF),
Ephrins Ephrin A1, Ephrin A2, Ephrin A3, Ephrin A4, Ephrin A5,
Ephrin B1, Ephrin B2, Ephrin B3, Erythropoietin (EPO), Fibroblast
growth factor (FGF), Foetal Bovine Somatotrophin (FBS), GDNF family
of ligands Glial cell line-derived neurotrophic factor (GDNF),
Neurturin, Persephin, Artemin, Growth differentiation factor-9
(GDF9), Hepatocyte growth factor (HGF), Hepatoma-derived growth
factor (HDGF), Insulin, Insulin-like growth factors Insulin-like,
growth factor-1 (IGF-1), Insulin-like growth factor-2 (IGF-2),
Interleukins IL-1-Cofactor for IL-3 and IL-6, IL-2, -3, -4, -5, -6,
-7, Keratinocyte growth factor (KGF), Migration-stimulating factor
(MSF), Macrophage-stimulating protein (MSP), also known as
hepatocyte growth factor-like protein (HGFLP), Myostatin (GDF-8),
Neuregulins Neuregulin 1 (NRG1), Neuregulin 2 (NRG2), Neuregulin 3
(NRG3), Neuregulin 4 (NRG4), Neurotrophins Brain-derived
neurotrophic factor (BDNF), Nerve growth factor (NGF),
Neurotrophin-3 (NT-3), Neurotrophin-4 (NT-4), Placental growth
factor (PGF), Platelet-derived growth factor (PDGF), Renalase
(RNLS)--Anti-apoptotic survival factor, T-cell growth factor
(TCGF), Thrombopoietin (TPO), Transforming growth factors
Transforming growth factor alpha (TGF-.alpha.), Transforming growth
factor beta (TGF-.beta.), Tumor necrosis factor-alpha
(TNF-.alpha.), Vascular endothelial growth factor (VEGF), Wnt
Signaling Pathway and receptors of the same.
[0171] More specifically, a number of drug targets for human
therapeutics represent protein targets to which protein target
moiety may be bound and incorporated into compounds according to
the present invention. These include proteins which may be used to
restore function in numerous polygenic diseases, including for
example B7.1 and B7, TINFR1m, TNFR2, NADPH oxidase, BclIBax and
other partners in the apotosis pathway, C5a receptor, HMG-CoA
reductase, PDE V phosphodiesterase type, PDE IV phosphodiesterase
type 4, PDE I, PDEII, PDEIII, squalene cyclase inhibitor, CXCR1,
CXCR2, nitric oxide (NO) synthase, cyclo-oxygenase 1,
cyclo-oxygenase 2, 5HT receptors, dopamine receptors, G Proteins,
i.e., Gq, histamine receptors, 5-lipoxygenase, tryptase serine
protease, thymidylate synthase, purine nucleoside phosphorylase,
GAPDH trypanosomal, glycogen phosphorylase, Carbonic anhydrase,
chemokine receptors, JAK, STAT, RXR and similar, HIV 1 protease,
HIV 1 integrase, influenza, neuramimidase, hepatitis B reverse
transcriptase, sodium channel, multi drug resistance (MDR), protein
P-glycoprotein (and MRP), tyrosine kinases, CD23, CD124, tyrosine
kinase p56 lck, CD4, CD5, IL-2 receptor, IL-1 receptor, TNF-alphaR,
ICAM1, Cat+ channels, VCAM, VLA-4 integrin, selectins, CD40/CD40L,
newokinins and receptors, inosine monophosphate dehydrogenase, p38
MAP Kinase, Ras, Raf, ERK pathway, FLT-3, KSR1, SMARCA, SMARCA2,
interleukin-1 converting enzyme, caspase, HCV, NS3 protease, HCV
NS3 RNA helicase, glycinamide ribonucleotide formyl transferase,
rhinovirus 3C protease, herpes simplex virus-1 (HSV-I), protease,
cytomegalovirus (CMV) protease, poly (ADP-ribose) polymerase,
cyclin dependent kinases, vascular endothelial growth factor,
oxytocin receptor, microsomal transfer protein inhibitor, bile acid
transport inhibitor, 5 alpha reductase inhibitors, angiotensin 11,
glycine receptor, noradrenaline reuptake receptor, endothelin
receptors, neuropeptide Y and receptor, estrogen receptors,
androgen receptors, adenosine receptors, adenosine kinase and AMP
deaminase, purinergic receptors (P2Y1, P2Y2, P2Y4, P2Y6, P2X1-7),
farnesyltransferases, geranylgeranyl transferase, TrkA a receptor
for NGF, beta-amyloid, tyrosine kinase Flk-IIKDR, vitronectin
receptor, integrin receptor, Her-21 neu, telomerase inhibition,
cytosolic phospholipaseA2 and EGF receptor tyrosine kinase.
Additional protein targets include, for example, ecdysone
20-monooxygenase, ion channel of the GABA gated chloride channel,
acetylcholinesterase, voltage-sensitive sodium channel protein,
calcium release channel, and chloride channels. Still further
target proteins include Acetyl-CoA carboxylase, adenylosuccinate
synthetase, protoporphyrinogen oxidase, and
enolpyruvylshikimate-phosphate synthase.
[0172] Haloalkane dehalogenase enzymes are another target of
specific compounds according to the present invention. Compounds
according to the present invention which contain chloroalkane
peptide binding moieties (C1-C12 often about C2-C10 alkyl halo
groups) may be used to inhibit and/or degrade haloalkane
dehalogenase enzymes which are used in fusion proteins or related
dioagnostic proteins as described in PCT/US2012/063401 filed Dec.
6, 2011 and published as WO 2012/078559 on Jun. 14, 2012, the
contents of which is incorporated by reference herein.
[0173] These various protein targets 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
invention, the level of activity of the protein may be altered for
therapeutic end result.
[0174] The compositions described below exemplify some of the
members of these types of small molecule target protein binding
moieties. Such small molecule target protein binding moieties also
include pharmaceutically acceptable salts, enantiomers, solvates
and polymorphs of these compositions, as well as other small
molecules that may target a protein of interest. References which
are cited hereinbelow are incorporated by reference herein in their
entirety.
[0175] I. Heat Shock Protein 90 (HSP90) Inhibitors:
[0176] HSP90 inhibitors as used herein include, but are not limited
to:
[0177] 1. The HSP90 inhibitors identified in Vallee, et al.,
"Tricyclic Series of Heat Shock Protein 90 (HSP90) Inhibitors Part
I: Discovery of Tricyclic Imidazo[4,5-C]Pyridines as Potent
Inhibitors of the HSP90 Molecular Chaperone (2011) J. Med. Chem.
54: 7206, including YKB
(N-[4-(3H-imidazo[4,5-C]Pyridin-2-yl)-9H-Fluoren-9-yl]-succinamide):
##STR00022##
derivatized where a linker group L or a -(L-MLM) group is attached,
for example, via the terminal amide group;
[0178] 2. The HSP90 inhibitor p54 (modified)
(8-[(2,4-dimethylphenyl)sulfanyl]-3]pent-4-yn-1-yl-3H-purin-6-amine):
##STR00023##
derivatized where a linker group L or a -(L-MLM) group is attached,
for example, via the terminal acetylene group;
[0179] 3. The HSP90 inhibitors (modified) identified in Brough, et
al., "4,5-Diarylisoxazole HSP90 Chaperone Inhibitors: Potential
Therapeutic Agents for the Treatment of Cancer", J. MED. CHEM. vol:
51, pag: 196 (2008), including the compound 2GJ
(5-[2,4-dihydroxy-5-(1-methylethyl)phenyl]-n-ethyl-4-[4-(morpholin-4-ylme-
thyl)phenyl]isoxazole-3-carboxamide) having the structure:
##STR00024##
derivatized, where a linker group L or a -(L-MLM) group is
attached, for example, via the amide group (at the amine or at the
alkyl group on the amine);
[0180] 4. The HSP90 inhibitors (modified) identified in Wright, et
al., Structure-Activity Relationships in Purine-Based Inhibitor
Binding to HSP90 Isoforms, Chem Biol. 2004 June; 11(6):775-85,
including the HSP90 inhibitor PU3 having the structure:
##STR00025##
derivatized where a linker group L or -(L-MLM) is attached, for
example, via the butyl group; and
[0181] 5. The HSP90 inhibitor geldanamycin
((4E,6Z,8S,9S,10E,12S,13R,14S,16R)-13-hydroxy-8,14,19-trimethoxy-4,10,12,-
16-tetramethyl-3,20,22-trioxo-2-azabicyclo[16.3.1] (derivatized) or
any of its derivatives (e.g.
17-alkylamino-17-desmethoxygeldanamycin ("17-AAG") or
17-(2-dimethylaminoethyl)amino-17-desmethoxygeldanamycin
("17-DMAG")) (derivatized, where a linker group L or a-(L-MLM)
group is attached, for example, via the amide group).
[0182] II. Kinase and Phosphatase Inhibitors:
[0183] Kinase inhibitors as used herein include, but are not
limited to:
[0184] 1. Erlotinib Derivative Tyrosine Kinase Inhibitor:
##STR00026##
where R is a linker group L or a -(L-MLM) group attached, for
example, via the ether group;
[0185] 2. The kinase inhibitor sunitinib (derivatized):
##STR00027##
derivatized where R is a linker group L or a -(L-MLM) group
attached, for example, to the pyrrole moiety;
[0186] 3. Kinase Inhibitor sorafenib (derivatized):
##STR00028##
derivatized where R is a linker group L or a -(L-MLM) group
attached, for example, to the amide moiety;
[0187] 4. The kinase inhibitor desatinib (derivatized):
##STR00029##
derivatized where R is a linker group L or a-(L-MLM) attached, for
example, to the pyrimidine;
[0188] 5. The kinase inhibitor lapatinib (derivatized):
##STR00030##
derivatized where a linker group L or a-(L-MLM) group is attached,
for example, via the terminal methyl of the sulfonyl methyl
group;
[0189] 6. The kinase inhibitor U09-CX-5279 (derivatized):
##STR00031##
derivatized where a linker group L or a -(L-MLM) group is attached,
for example, via the amine (aniline), carboxylic acid or amine
alpha to cyclopropyl group, or cyclopropyl group;
[0190] 7. The kinase inhibitors identified in Millan, et al.,
Design and Synthesis of Inhaled P38 Inhibitors for the Treatment of
Chronic Obstructive Pulmonary Disease, J. MED. CHEM. vol: 54, pag:
7797 (2011), including the kinase inhibitors Y1W and Y1X
(Derivatized) having the structures:
##STR00032##
[0191]
YIX(1-ethyl-3-(2-{[3-(1-methylethyl)[1,2,4]triazolo[4,3-a]pyridine--
6-yl]sulfanyl}benzyl)urea, derivatized where a linker group L or
a-(L-MLM) group is attached, for example, via the .sup.ipropyl
group;
##STR00033##
derivatized where a linker group L or a -(L-MLM) group is attached,
for example, preferably via either the i-propyl group or the
t-butyl group;
[0192] 8. The kinase inhibitors identified in Schenkel, et al.,
Discovery of Potent and Highly Selective Thienopyridine Janus
Kinase 2 Inhibitors J. Med. Chem., 2011, 54 (24), pp 8440-8450,
including the compounds 6TP and 0TP (Derivatized) having the
structures:
##STR00034##
derivatized where a linker group L or a -(L-MLM) group is attached,
for example, via the terminal methyl group bound to amide
moiety;
##STR00035##
derivatized where a linker group L or a -(L-MLM)group is attached,
for example, via the terminal methyl group bound to the amide
moiety;
[0193] 9. The kinase inhibitors identified in Van Eis, et al.,
"2,6-Naphthyridines as potent and selective inhibitors of the novel
protein kinase C isozymes", Biorg. Med. Chem. Lett. 2011 Dec. 15;
21(24):7367-72, including the kinase inhibitor 07U having the
structure:
##STR00036##
derivatized where a linker group L or a -(L-MLM)group is attached,
for example, via the secondary amine or terminal amino group;
[0194] 10. The kinase inhibitors identified in Lountos, et al.,
"Structural Characterization of Inhibitor Complexes with Checkpoint
Kinase 2 (Chk2), a Drug Target for Cancer Therapy", J. STRUCT.
BIOL. vol: 176, pag: 292 (2011), including the kinase inhibitor YCF
having the structure:
##STR00037##
derivatized where a linker group L or a -(L-MLM) group is attached,
for example, via either of the terminal hydroxyl groups;
[0195] 11. The kinase inhibitors identified in Lountos, et al.,
"Structural Characterization of Inhibitor Complexes with Checkpoint
Kinase 2 (Chk2), a Drug Target for Cancer Therapy", J. STRUCT.
BIOL. vol: 176, pag: 292 (2011), including the kinase inhibitors
XK9 and NXP (derivatized) having the structures:
##STR00038##
derivatized where a linker group L or a -(L-MLM) group is attached,
for example, via the terminal hydroxyl group (XK9) or the hydrazone
group (NXP);
[0196] 12. The kinase inhibitor afatinib (derivatized)
(N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[[(3S)-tetrahydro-3-furanyl]oxy]-
-6-quinazolinyl]-4(dimethylamino)-2-butenamide) (Derivatized where
a linker group L or a -(L-MLM) group is attached, for example, via
the aliphatic amine group);
[0197] 13. The kinase inhibitor fostamatinib (derivatized)
([6-({5-fluoro-2-[(3,4,5-trimethoxyphenyl)amino]pyrimidin-4-yl}amino)-2,2-
-dimethyl-3-oxo-2,3-dihydro-4H-pyrido[3,2-b]-1,4-oxazin-4-yl]methyl
disodium phosphate hexahydrate) (Derivatized where a linker group L
or a -(L-MLM) group is attached, for example, via a methoxy
group);
[0198] 14. The kinase inhibitor gefitinib (derivatized)
(N-(3-chloro-4-fluoro-phenyl)-7-methoxy-6-(3-morpholin-4-ylpropoxy)quinaz-
olin-4-amine):
##STR00039##
derivatized where a linker group L or a -(L-MLM) group is attached,
for example, via a methoxy or ether group;
[0199] 15. The kinase inhibitor lenvatinib (derivatized)
(4-[3-chloro-4-(cyclopropylcarbamoylamino)phenoxy]-7-methoxy-quinoline-6--
carboxamide) (derivatized where a linker group L or a -(L-MLM)
group is attached, for example, via the cyclopropyl group);
[0200] 16. The kinase inhibitor vandetanib (derivatized)
(N-(4-bromo-2-fluorophenyl)-6-methoxy-7-[(1-methylpiperidin-4-yl)methoxy]-
quinazolin-4-amine) (derivatized where a linker group L or a
-(L-MLM) group is attached, for example, via the methoxy or
hydroxyl group);
[0201] 17. The kinase inhibitor vemurafenib (derivatized)
(propane-1-sulfonic acid
{3-[5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4-difluoro-
-phenyl}-amide), derivatized where a linker group L or a -(L-MLM)
group is attached, for example, via the sulfonyl propyl group;
[0202] 18. The kinase inhibitor Gleevec (derivatized):
##STR00040##
derivatized where R as a linker group L or a-(L-MLM) group is
attached, for example, via the amide group or via the aniline amine
group;
[0203] 19. The kinase inhibitor pazopanib (derivatized) (VEGFR3
inhibitor):
##STR00041##
derivatized where R is a linker group L or a -(L-MLM) group
attached, for example, to the phenyl moiety or via the aniline
amine group;
[0204] 20. The kinase inhibitor AT-9283 (Derivatized) Aurora Kinase
Inhibitor
##STR00042##
where R is a linker group L or a -(L-MLM) group attached, for
example, to the phenyl moiety);
[0205] 21. The kinase inhibitor TAE684 (derivatized) ALK
inhibitor
##STR00043##
where R is a linker group L or a -(L-MLM) group attached, for
example, to the phenyl moiety);
[0206] 22. The kinase inhibitor nilotanib (derivatized) Abl
inhibitor:
##STR00044##
derivatized where R is a linker group L or a -(L-MLM) group
attached, for example, to the phenyl moiety or the aniline amine
group;
[0207] 23. Kinase Inhibitor NVP-BSK805 (derivatized) JAK2
Inhibitor
##STR00045##
derivatized where R is a linker group L or a -(L-MLM) group
attached, for example, to the phenyl moiety or the diazole
group;
[0208] 24. Kinase Inhibitor crizotinib Derivatized Alk
Inhibitor
##STR00046##
derivatized where R is a linker group L or a -(L-MLM) group
attached, for example, to the phenyl moiety or the diazole
group;
[0209] 25. Kinase Inhibitor JNJ FMS (derivatized) Inhibitor
##STR00047##
derivatized where R is a linker group L or a -(L-MLM) group
attached, for example, to the phenyl moiety;
[0210] 26. The kinase inhibitor foretinib (derivatized) Met
Inhibitor
##STR00048##
derivatized where R is a linker group L or a -(L-MLM)group
attached, for example, to the phenyl moiety or a hydroxyl or ether
group on the quinoline moiety;
[0211] 27. The allosteric Protein Tyrosine Phosphatase Inhibitor
PTP1B (derivatized):
##STR00049##
derivatized where a linker group L or a -(L-MLM) group is attached,
for example, at R, as indicated;
[0212] 28. The inhibitor of SHP-2 Domain of Tyrosine Phosphatase
(derivatized):
##STR00050##
derivatized where a linker group L or a -(L-MLM) group is attached,
for example, at R;
[0213] 29. The inhibitor (derivatized) of BRAF
(BRAF.sup.V600E)/MEK:
##STR00051##
derivatized where a linker group L or a-(L-MLM) group is attached,
for example, at R;
[0214] 30. Inhibitor (derivatized) of Tyrosine Kinase ABL
##STR00052##
derivatized where a linker group L or a-(L-MLM) group is attached,
for example, at R;
[0215] 31. The kinase inhibitor OSI-027 (derivatized) mTORC1/2
inhibitor
##STR00053##
derivatized where a linker group L or a-(L-MLM) group is attached,
for example, at R;
[0216] 32. The kinase inhibitor OSI-930 (derivatized) c-Kit/KDR
inhibitor
##STR00054##
derivatized where a linker group L or a-(L-MLM) group is attached,
for example, at R; and
[0217] 33. The kinase inhibitor OSI-906 (derivatized) IGF1R/IR
inhibitor
##STR00055##
derivatized where a linker group L or a-(L-MLM) group is attached,
for example, at R.
[0218] Wherein, in any of the embodiments described in sections
I-XVII, "R" designates a site for attachment of a linker group L or
a -(L-MLM)group on the piperazine moiety.
[0219] III. HDM2/MDM2 Inhibitors:
[0220] HDM2/MDM2 inhibitors as used herein include, but are not
limited to:
[0221] 1. 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:
##STR00056##
(derivatized where a linker group L or a -(L-MLM)group is attached,
for example, at the methoxy group or as a hydroxyl group);
##STR00057##
(derivatized where a linker group L or a -(L-MLM) group is
attached, for example, at the methoxy group or hydroxyl group);
##STR00058##
(derivatized where a linker group L or a -(L-MLM) group is
attached, for example, via the methoxy group or as a hydroxyl
group); and
[0222] 2. Trans-4-Iodo-4'-Boranyl-Chalcone
##STR00059##
[0223] (derivatized where a linker group L or a a linker group L or
a-(L-MLM) group is attached, for example, via a hydroxy group).
[0224] IV. Compounds Targeting Human BET Bromodomain-Containing
Proteins:
[0225] In certain embodiments, "PTM" can be ligands binding to
Bromo- and Extra-terminal (BET) proteins BRD2, BRD3 and BRD4.
Compounds targeting Human BET Bromodomain-containing proteins
include, but are not limited to the compounds associated with the
targets as described below, where "R" or "linker" designates a site
for linker group L or a-(L-MLM) group attachment, for example:
[0226] 1. JQ1, Filippakopoulos et al. Selective inhibition of BET
bromodomains. Nature (2010):
##STR00060## ##STR00061##
[0227] 2. I-BET, Nicodeme et al. Supression of Inflammation by a
Synthetic Histone Mimic. Nature (2010). Chung et al. Discovery and
Characterization of Small Molecule Inhibitors of the BET Family
Bromodomains. J. Med Chem. (2011):
##STR00062##
[0228] 3. Compounds described in Hewings et al.
3,5-Dimethylisoxazoles Act as Acetyl-lysine Bromodomain Ligands. J.
Med. Chem. (2011) 54 6761-6770.
##STR00063##
[0229] 4. I-BET151, Dawson et al. Inhibition of BET Recruitment to
Chromatin as an Efective Treatment for MLL-fusion Leukemia. Nature
(2011):
##STR00064##
[0230] 5. Carbazole type (US 2015/0256700)
##STR00065##
[0231] 6. Pyrrolopyridone type (US 2015/0148342)
##STR00066##
[0232] 7. Tetrahydroquinoline type (WO 2015/074064)
##STR00067##
[0233] 8. Triazolopyrazine type (WO 2015/067770)
##STR00068##
[0234] 9. Pyridone type (WO 2015/022332)
##STR00069##
[0235] 10. Quinazolinone type (WO 2015/015318)
##STR00070##
[0236] 11. Dihydropyridopyrazinone type (WO 2015/011084)
##STR00071##
[0237] V. HDAC Inhibitors:
[0238] HDAC Inhibitors (derivatized) include, but are not limited
to:
[0239] 1. Finnin, M. S. et al. Structures of Histone Deacetylase
Homologue Bound to the TSA and SAHA Inhibitors. Nature 40, 188-193
(1999).
##STR00072##
(Derivatized where "R" designates a site for attachment, for
example, of a linker group L or a -(L-MLM) group); and
[0240] 2. Compounds as defined by formula (I) of PCT WO0222577
("DEACETYLASE INHIBITORS") (Derivatized where a linker group L or a
-(L-MLM) group is attached, for example, via the hydroxyl
group);
[0241] VI. Human Lysine Methyltransferase Inhibitors:
[0242] Human Lysine Methyltransferase inhibitors include, but are
not limited to:
[0243] 1. Chang et al. Structural Basis for G9a-Like protein Lysine
Methyltransferase Inhibition by BIX-1294. Nat. Struct. Biol. (2009)
16(3) 312.
##STR00073##
[0244] (Derivatized where "R" designates a site for attachment, for
example, of a linker group L or a -(L-MLM) group);
[0245] 2. Liu, F. et al Discovery of a
2,4-Diamino-7-aminoalkoxyquinazoline as a Potent and Selective
Inhibitor of Histone Methyltransferase G9a. J. Med. Chem. (2009)
52(24) 7950.
##STR00074##
[0246] (Derivatized where "R" designates a potential site for
attachment, for example, of a linker group L or a -(L-MLM)
group);
[0247] 3. Azacitidine (derivatized)
(4-amino-1-.beta.-D-ribofuranosyl-1,3,5-triazin-2(1H)-one)
(Derivatized where a linker group L or a -(L-MLM) group is
attached, for example, via the hydroxy or amino groups); and
[0248] 4. Decitabine (derivatized)
(4-amino-1-(2-deoxy-b-D-erythro-pentofuranosyl)-1, 3,
5-triazin-2(1H)-one) (Derivatized where a linker group L or a
-(L-MLM) group is attached, for example, via either of the hydroxy
groups or at the amino group).
[0249] VII. Angiogenesis Inhibitors:
[0250] Angiogenesis inhibitors include, but are not limited to:
[0251] 1. GA-1 (derivatized) and derivatives and analogs thereof,
having the structure(s) and binding to linkers as described in
Sakamoto, et al., Development of Protacs to target cancer-promoting
proteins for ubiquitination and degradation, Mol Cell Proteomics
2003 December; 2(12):1350-8;
[0252] 2. Estradiol (derivatized), which may be bound to a linker
group L or a -(L-MLM) group as is generally described in
Rodriguez-Gonzalez, et al., Targeting steroid hormone receptors for
ubiquitination and degradation in breast and prostate cancer,
Oncogene (2008) 27, 7201-7211;
[0253] 3. Estradiol, testosterone (derivatized) and related
derivatives, including but not limited to DHT and derivatives and
analogs thereof, having the structure(s) and binding to a linker
group L or a -(L-MLM) group as generally described in Sakamoto, et
al., Development of Protacs to target cancer-promoting proteins for
ubiquitination and degradation, Mol Cell Proteomics 2003 December;
2(12):1350-8; and
[0254] 4. Ovalicin, fumagillin (derivatized), and derivatives and
analogs thereof, having the structure(s) and binding to a linker
group L or a -(L-MLM) group as is generally described in Sakamoto,
et al., Protacs: chimeric molecules that target proteins to the
Skp1-Cullin-F box complex for ubiquitination and degradation Proc
Natl Acad Sci USA. 2001 Jul. 17; 98(15):8554-9 and U.S. Pat. No.
7,208,157.
[0255] VIII. Immunosuppressive Compounds:
[0256] Immunosuppressive compounds include, but are not limited
to:
[0257] 1. AP21998 (derivatized), having the structure(s) and
binding to a linker group L or a -(L-MLM) group as is generally
described in Schneekloth, et al., Chemical Genetic Control of
Protein Levels: Selective in Vivo Targeted Degradation, J. AM.
CHEM. SOC. 2004, 126, 3748-3754;
[0258] 2. Glucocorticoids (e.g., hydrocortisone, prednisone,
prednisolone, and methylprednisolone) (Derivatized where a linker
group L or a -(L-MLM) group is to bound, e.g. to any of the
hydroxyls) and beclometasone dipropionate (Derivatized where a
linker group or a -(L-MLM) is bound, e.g. to a proprionate);
[0259] 3. Methotrexate (Derivatized where a linker group or a
-(L-MLM) group can be bound, e.g. to either of the terminal
hydroxyls);
[0260] 4. Ciclosporin (Derivatized where a linker group or a
-(L-MLM) group can be bound, e.g. at any of the butyl groups);
[0261] 5. Tacrolimus (FK-506) and rapamycin (Derivatized where a
linker group L or a -(L-MLM) group can be bound, e.g. at one of the
methoxy groups); and
[0262] 6. Actinomycins (Derivatized where a linker group L or a
-(L-MLM) group can be bound, e.g. at one of the isopropyl
groups).
[0263] IX. Compounds Targeting the Aryl Hydrocarbon Receptor
(AHR):
[0264] Compounds targeting the aryl hydrocarbon receptor (AHR)
include, but are not limited to:
[0265] 1. Apigenin (Derivatized in a way which binds to a linker
group L or a -(L-MLM) group as is generally illustrated in Lee, et
al., Targeted Degradation of the Aryl Hydrocarbon Receptor by the
PROTAC Approach: A Useful Chemical Genetic Tool, ChemBioChem Volume
8, Issue 17, pages 2058-2062, Nov. 23, 2007); and
[0266] 2. SR1 and LGC006 (derivatized such that a linker group L or
a -(L-MLM) is bound), as described in Boitano, et al., Aryl
Hydrocarbon Receptor Antagonists Promote the Expansion of Human
Hematopoietic Stem Cells, Science 10 Sep. 2010: Vol. 329 no. 5997
pp. 1345-1348.
[0267] X. Compounds Targeting RAF Receptor (Kinase):
##STR00075##
[0268] (Derivatized where "R" designates a site for linker group L
or -(L-MLM) group attachment, for example).
[0269] XI. Compounds Targeting FKBP:
##STR00076##
[0270] (Derivatized where "R" designates a site for a linker group
L or a -(L-MLM) group attachment, for example).
[0271] XII. Compounds Targeting Androgen Receptor (AR)
[0272] 1. RU59063 Ligand (derivatized) of Androgen Rceptor
##STR00077##
[0273] (Derivatized where "R" designates a site for a linker group
L or a -(L-MLM) group attachment, for example).
[0274] 2. SARM Ligand (derivatized) of Androgen Receptor
##STR00078##
[0275] (Derivatized where "R" designates a site for a linker group
L or a-(L-MLM) group attachment, for example).
[0276] 3. Androgen Receptor Ligand DHT (derivatized)
##STR00079##
[0277] (Derivatized where "R" designates a site for a linker group
L or -(L-MLM) group attachment, for example).
[0278] 4. MDV3100 Ligand (derivatized)
##STR00080##
[0279] 5. ARN-509 Ligand (derivatized)
##STR00081##
[0280] 6. Hexahydrobenzisoxazoles
##STR00082##
[0281] 7. Tetramethylcyclobutanes
##STR00083##
[0282] XIII. Compounds Targeting Estrogen Receptor (ER)
ICI-182780
[0283] 1. Estrogen Receptor Ligand
##STR00084##
[0284] (Derivatized where "R" designates a site for linker group L
or -(L-MLM) group attachment).
[0285] XIV. Compounds Targeting Thyroid Hormone Receptor (TR)
[0286] 1. Thyroid Hormone Receptor Ligand (derivatized)
##STR00085##
[0287] (Derivatized where "R" designates a site for linker group L
or -(L-MLM) group attachment and MOMO indicates a methoxymethoxy
group).
[0288] XV. Compounds Targeting HIV Protease
[0289] 1. Inhibitor of HIV Protease (derivatized)
##STR00086##
[0290] (Derivatized where "R" designates a site for linker group L
or -(L-MLM) group attachment). See, J. Med. Chem. 2010, 53,
521-538.
[0291] 2. Inhibitor of HIV Protease
##STR00087##
[0292] (Derivatized where "R" designates a potential site for
linker group L or -(L-MLM) group attachment). See, J. Med. Chem.
2010, 53, 521-538.
[0293] XVI. Compounds Targeting HIV Integrase
[0294] 1. Inhibitor of HIV Integrase (derivatized)
##STR00088##
[0295] (Derivatized where "R" designates a site for linker group L
or -(L-MLM) group attachment). See, J. Med. Chem. 2010, 53,
6466.
[0296] 2. Inhibitor of HIV Integrase (derivatized)
##STR00089##
[0297] 3. Inhibitor of HIV integrase Isetntress (derivatized)
##STR00090##
[0298] (Derivatized where "R" designates a site for linker group L
or -(L-MLM) group attachment). See, J. Med. Chem. 2010, 53,
6466.
[0299] XVII. Compounds Targeting HCV Protease
[0300] 1. Inhibitors of HCV Protease (derivatized)
##STR00091##
[0301] (Derivatized where "R" designates a site for linker group L
or -(L-MLM) group attachment).
[0302] XVIII. Compounds Targeting Acyl-Protein Thioesterase-1 and
-2 (APT1 and APT2)
[0303] 1. Inhibitor of APT1 and APT2 (derivatized)
##STR00092##
[0304] (Derivatized where "R" designates a site for linker group L
or -(L-MLM) group attachment). See, Angew. Chem. Int. Ed. 2011, 50,
9838-9842, where L is a linker group as otherwise described herein
and said MLM group is as otherwise described herein such that
-(L-MLM) binds the MLM group to a PTM group as otherwise described
herein.
[0305] Therapeutic Compositions
[0306] 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.
[0307] 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.
[0308] 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.
[0309] 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 disclosureion 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.
[0310] 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.
[0311] 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.
[0312] 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.
[0313] 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.
[0314] 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.
[0315] 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.
[0316] 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 invention
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 invention, 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.
[0317] 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.
[0318] 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.
[0319] 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.
[0320] 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 invention.
[0321] 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.
[0322] 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 invention 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.
[0323] 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.
[0324] 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.
[0325] 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.
[0326] 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.
[0327] 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.
[0328] 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.
[0329] 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.
[0330] 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.
[0331] 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 invention, one or more compounds according
to the present invention are coadministered with another bioactive
agent, such as an erythropoietin stimulating agent or a would
healing agent, including an antibiotic, as otherwise described
herein.
[0332] 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.
[0333] If administered intravenously, preferred carriers are
physiological saline or phosphate buffered saline (PBS).
[0334] 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.
[0335] 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 are 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.
[0336] Therapeutic Methods
[0337] 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.
[0338] 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, including cancer, which may be treated using
compounds according to the present invention are set forth
hereinabove.
[0339] The description provides therapeutic compositions as
described herein for effectuating the degradation of proteins of
interest for the treatment or amelioration of a disease, e.g.,
cancer. In certain additional embodiments, the disease is multiple
myeloma. As such, in another aspect, the description provides a
method of ubiquitinating/degrading a target protein in a cell. In
certain embodiments, the method comprises administering a
bifunctional compound as described herein comprising, e.g., a MLM
and a PTM, preferably linked through a linker moiety, as otherwise
described herein, wherein the MLM is coupled to the PTM and wherein
the MLM recognizes a ubiquitin pathway protein (e.g., an ubiquitin
ligase, preferably an E3 ubiquitin ligase such as, e.g., cereblon)
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 invention 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 pharamaceutically acceptable excipient, carrier,
adjuvant, another bioactive agent or combination thereof.
[0340] In additional embodiments, the description provides methods
for treating or emeliorating 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.
[0341] 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
invention.
[0342] In another embodiment, the present invention is directed to
a method of treating a human patient in need for a disease state or
condition modulated through a protein where the degradation of that
protein will produce a therapeutic effect in that patient, the
method comprising administering to a patient in need an effective
amount of a compound according to the present invention, optionally
in combination with another bioactive agent. 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
[0343] The term "disease state or condition" is used to describe
any disease state or condition wherein protein dysregulation (i.e.,
the amount of protein expressed in a patient is elevated) occurs
and where degradation of one or more 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.
[0344] Disease states of conditions which may be treated using
compounds according to the present invention include, for example,
asthma, autoimmune diseases such as multiple sclerosis, various
cancers, ciliopathies, cleft palate, diabetes, heart disease,
hypertension, inflammatory bowel disease, mental retardation, mood
disorder, obesity, refractive error, infertility, Angelman
syndrome, Canavan disease, Coeliac disease, Charcot-Marie-Tooth
disease, Cystic fibrosis, Duchenne muscular dystrophy,
Haemochromatosis, Haemophilia, Klinefelter's syndrome,
Neurofibromatosis, Phenylketonuria, Polycystic kidney disease,
(PKD1) or 4 (PKD2) Prader-Willi syndrome, Sickle-cell disease,
Tay-Sachs disease, Turner syndrome.
[0345] Further disease states or conditions which may be treated by
compounds according to the present invention include Alzheimer's
disease, Amyotrophic lateral sclerosis (Lou Gehrig's disease),
Anorexia nervosa, Anxiety disorder, Atherosclerosis, Attention
deficit hyperactivity disorder, Autism, Bipolar disorder, Chronic
fatigue syndrome, Chronic obstructive pulmonary disease, Crohn's
disease, Coronary heart disease, Dementia, Depression, Diabetes
mellitus type 1, Diabetes mellitus type 2, Epilepsy, Guillain-Barre
syndrome, Irritable bowel syndrome, Lupus, Metabolic syndrome,
Multiple sclerosis, Myocardial infarction, Obesity,
Obsessive-compulsive disorder, Panic disorder, Parkinson's disease,
Psoriasis, Rheumatoid arthritis, Sarcoidosis, Schizophrenia,
Stroke, Thromboangiitis obliterans, Tourette syndrome,
Vasculitis.
[0346] Still additional disease states or conditions which can be
treated by compounds according to the present invention include
aceruloplasminemia, Achondrogenesis type II, achondroplasia,
Acrocephaly, Gaucher disease type 2, acute intermittent porphyria,
Canavan disease, Adenomatous Polyposis Coli, ALA dehydratase
deficiency, adenylosuccinate lyase deficiency, Adrenogenital
syndrome, Adrenoleukodystrophy, ALA-D porphyria, ALA dehydratase
deficiency, Alkaptonuria, Alexander disease, Alkaptonuric
ochronosis, alpha 1-antitrypsin deficiency, alpha-1 proteinase
inhibitor, emphysema, amyotrophic lateral sclerosis Alstrom
syndrome, Alexander disease, Amelogenesis imperfecta, ALA
dehydratase deficiency, Anderson-Fabry disease, androgen
insensitivity syndrome, Anemia Angiokeratoma Corporis Diffusum,
Angiomatosis retinae (von Hippel-Lindau disease) Apert syndrome,
Arachnodactyly (Marfan syndrome), Stickler syndrome, Arthrochalasis
multiplex congenital (Ehlers-Danlos syndrome#arthrochalasia type)
ataxia telangiectasia, Rett syndrome, primary pulmonary
hypertension, Sandhoff disease, neurofibromatosis type II,
Beare-Stevenson cutis gyrata syndrome, Mediterranean fever,
familial, Benjamin syndrome, beta-thalassemia, Bilateral Acoustic
Neurofibromatosis (neurofibromatosis type II), factor V Leiden
thrombophilia, Bloch-Sulzberger syndrome (incontinentia pigmenti),
Bloom syndrome, X-linked sideroblastic anemia, Bonnevie-Ullrich
syndrome (Turner syndrome), Bourneville disease (tuberous
sclerosis), prion disease, Birt-Hogg-Dube syndrome, Brittle bone
disease (osteogenesis imperfecta), Broad Thumb-Hallux syndrome
(Rubinstein-Taybi syndrome), Bronze Diabetes/Bronzed Cirrhosis
(hemochromatosis), Bulbospinal muscular atrophy (Kennedy's
disease), Burger-Grutz syndrome (lipoprotein lipase deficiency),
CGD Chronic granulomatous disorder, Campomelic dysplasia,
biotinidase deficiency, Cardiomyopathy (Noonan syndrome), Cri du
chat, CAVD (congenital absence of the vas deferens), Caylor
cardiofacial syndrome (CBAVD), CEP (congenital erythropoietic
porphyria), cystic fibrosis, congenital hypothyroidism,
Chondrodystrophy syndrome (achondroplasia),
otospondylomegaepiphyseal dysplasia, Lesch-Nyhan syndrome,
galactosemia, Ehlers-Danlos syndrome, Thanatophoric dysplasia,
Coffin-Lowry syndrome, Cockayne syndrome, (familial adenomatous
polyposis), Congenital erythropoietic porphyria, Congenital heart
disease, Methemoglobinemia/Congenital methaemoglobinaemia,
achondroplasia, X-linked sideroblastic anemia, Connective tissue
disease, Conotruncal anomaly face syndrome, Cooley's Anemia
(beta-thalassemia), Copper storage disease (Wilson's disease),
Copper transport disease (Menkes disease), hereditary
coproporphyria, Cowden syndrome, Craniofacial dysarthrosis (Crouzon
syndrome), Creutzfeldt-Jakob disease (prion disease), Cockayne
syndrome, Cowden syndrome, Curschmann-Batten-Steinert syndrome
(myotonic dystrophy), Beare-Stevenson cutis gyrata syndrome,
primary hyperoxaluria, spondyloepimetaphyseal dysplasia (Strudwick
type), muscular dystrophy, Duchenne and Becker types (DBMD), Usher
syndrome, Degenerative nerve diseases including de Grouchy syndrome
and Dejerine-Sottas syndrome, developmental disabilities, distal
spinal muscular atrophy, type V, androgen insensitivity syndrome,
Diffuse Globoid Body Sclerosis (Krabbe disease), Di George's
syndrome, Dihydrotestosterone receptor deficiency, androgen
insensitivity syndrome, Down syndrome, Dwarfism, erythropoietic
protoporphyria Erythroid 5-aminolevulinate synthetase deficiency,
Erythropoietic porphyria, erythropoietic protoporphyria,
erythropoietic uroporphyria, Friedreich's ataxia, familial
paroxysmal polyserositis, porphyria cutanea tarda, familial
pressure sensitive neuropathy, primary pulmonary hypertension
(PPH), Fibrocystic disease of the pancreas, fragile X syndrome,
galactosemia, genetic brain disorders, Giant cell hepatitis
(Neonatal hemochromatosis), Gronblad-Strandberg syndrome
(pseudoxanthoma elasticum), Gunther disease (congenital
erythropoietic porphyria), haemochromatosis, Hallgren syndrome,
sickle cell anemia, hemophilia, hepatoerythropoietic porphyria
(HEP), Hippel-Lindau disease (von Hippel-Lindau disease),
Huntington's disease, Hutchinson-Gilford progeria syndrome
(progeria), Hyperandrogenism, Hypochondroplasia, Hypochromic
anemia, Immune system disorders, including X-linked severe combined
immunodeficiency, Insley-Astley syndrome, Jackson-Weiss syndrome,
Joubert syndrome, Lesch-Nyhan syndrome, Jackson-Weiss syndrome,
Kidney diseases, including hyperoxaluria, Klinefelter's syndrome,
Kniest dysplasia, Lacunar dementia, Langer-Saldino achondrogenesis,
ataxia telangiectasia, Lynch syndrome, Lysyl-hydroxylase
deficiency, Machado-Joseph disease, Metabolic disorders, including
Kniest dysplasia, Marfan syndrome, Movement disorders, Mowat-Wilson
syndrome, cystic fibrosis, Muenke syndrome, Multiple
neurofibromatosis, Nance-Insley syndrome, Nance-Sweeney
chondrodysplasia, Niemann-Pick disease, Noack syndrome (Pfeiffer
syndrome), Osler-Weber-Rendu disease, Peutz-Jeghers syndrome,
Polycystic kidney disease, polyostotic fibrous dysplasia
(McCune-Albright syndrome), Peutz-Jeghers syndrome,
Prader-Labhart-Willi syndrome, hemochromatosis, primary
hyperuricemia syndrome (Lesch-Nyhan syndrome), primary pulmonary
hypertension, primary senile degenerative dementia, prion disease,
progeria (Hutchinson Gilford Progeria Syndrome), progressive
chorea, chronic hereditary (Huntington) (Huntington's disease),
progressive muscular atrophy, spinal muscular atrophy, propionic
acidemia, protoporphyria, proximal myotonic dystrophy, pulmonary
arterial hypertension, PXE (pseudoxanthoma elasticum), Rb
(retinoblastoma), Recklinghausen disease (neurofibromatosis type
I), Recurrent polyserositis, Retinal disorders, Retinoblastoma,
Rett syndrome, RFALS type 3, Ricker syndrome, Riley-Day syndrome,
Roussy-Levy syndrome, severe achondroplasia with developmental
delay and acanthosis nigricans (SADDAN), Li-Fraumeni syndrome,
sarcoma, breast, leukemia, and adrenal gland (SBLA) syndrome,
sclerosis tuberose (tuberous sclerosis), SDAT, SED congenital
(spondyloepiphyseal dysplasia congenita), SED Strudwick
(spondyloepimetaphyseal dysplasia, Strudwick type), SEDc
(spondyloepiphyseal dysplasia congenita) SEMD, Strudwick type
(spondyloepimetaphyseal dysplasia, Strudwick type), Shprintzen
syndrome, Skin pigmentation disorders, Smith-Lemli-Opitz syndrome,
South-African genetic porphyria (variegate porphyria),
infantile-onset ascending hereditary spastic paralysis, Speech and
communication disorders, sphingolipidosis, Tay-Sachs disease,
spinocerebellar ataxia, Stickler syndrome, stroke, androgen
insensitivity syndrome, tetrahydrobiopterin deficiency,
beta-thalassemia, Thyroid disease, Tomaculous neuropathy
(hereditary neuropathy with liability to pressure palsies),
Treacher Collins syndrome, Triplo X syndrome (triple X syndrome),
Trisomy 21 (Down syndrome), Trisomy X, VHL syndrome (von
Hippel-Lindau disease), Vision impairment and blindness (Alstrom
syndrome), Vrolik disease, Waardenburg syndrome, Warburg Sjo
Fledelius Syndrome, Weis senbacher-Zweymuller syndrome,
Wolf-Hirschhorn syndrome, Wolff Periodic disease, Weis
senbacher-Zweymuller syndrome and Xeroderma pigmentosum, among
others.
[0347] The term "neoplasia" or "cancer" is used throughout the
specification to refer to the pathological process that results in
the formation and growth of neoplasm, i.e., abnormal tissue that
grows by cellular proliferation, often more rapidly than normal and
continues to grow after the stimuli that initiated the new growth
cease. Malignant neoplasms show partial or complete lack of
structural organization and functional coordination with the normal
tissue and most invade surrounding tissues, metastasize to several
sites, and are likely to recur after attempted removal and to cause
the death of the patient unless adequately treated. As used herein,
the term neoplasia is used to describe all cancerous disease states
and embraces or encompasses the pathological process associated
with malignant hematogenous, ascitic and solid tumors. Exemplary
cancers which may be treated by the present compounds either alone
or in combination with at least one additional anti-cancer agent
include squamous-cell carcinoma, basal cell carcinoma,
adenocarcinoma, hepatocellular carcinomas, and renal cell
carcinomas, cancer of the bladder, bowel, breast, cervix, colon,
esophagus, head, kidney, liver, lung, neck, ovary, pancreas,
prostate, and stomach; leukemias; benign and malignant lymphomas,
particularly Burkitt's lymphoma and Non-Hodgkin's lymphoma; benign
and malignant melanomas; myeloproliferative diseases; sarcomas,
including Ewing's sarcoma, hemangiosarcoma, Kaposi's sarcoma,
liposarcoma, myosarcomas, peripheral neuroepithelioma, synovial
sarcoma, gliomas, astrocytomas, oligodendrogliomas, ependymomas,
gliobastomas, neuroblastomas, ganglioneuromas, gangliogliomas,
medulloblastomas, pineal cell tumors, meningiomas, meningeal
sarcomas, neurofibromas, and Schwannomas; bowel cancer, breast
cancer, prostate cancer, cervical cancer, uterine cancer, lung
cancer, ovarian cancer, testicular cancer, thyroid cancer,
astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer,
liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's
disease, Wilms' tumor and teratocarcinomas. Additional cancers
which may be treated using compounds according to the present
invention include, for example, T-lineage Acute lymphoblastic
Leukemia (T-ALL), T-lineage lymphoblastic Lymphoma (T-LL),
Peripheral T-cell lymphoma, Adult T-cell Leukemia, Pre-B ALL, Pre-B
Lymphomas, Large B-cell Lymphoma, Burkitts Lymphoma, B-cell ALL,
Philadelphia chromosome positive ALL and Philadelphia chromosome
positive CML.
[0348] The term "bioactive agent" is used to describe an agent,
other than a compound according to the present invention, 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.
[0349] The term "anti-cancer agent" is used to describe an
anti-cancer agent, which may be incorporated into the bifunctional
compounds according to the present invention or incombination with
the same 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.
[0350] The term "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 invention), protease inhibitors,
fusion inhibitors, among others, exemplary compounds of which may
include, for example, 3TC (Lamivudine), AZT (Zidovudine), (-)-FTC,
ddl (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.
[0351] Other anti-HIV agents which may be used in coadministration
with compounds according to the present invention include, for
example, other NNRTI's (i.e., other than the NNRTI's according to
the present invention) 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 (TMC 125), 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.
[0352] 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 gastic
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 invention.
[0353] 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
[0354] The following structures are ligands for BET (bromodomain
and extra terminal domain). These ligands are used as an example
only to demonstrate the current invention of using MDM2 E3 ligase
to degrade a target protein and in no way limit the present
invention. In the exampled structures below, the target protein is
BRD2 (BRD2, BRD3 and BRD4). In a certain embodiment, the PTM is
selected from the group consisting of:
##STR00093## ##STR00094##
wherein "*" indicates one of the positions to connect linker.
[0355] The following are examples of androgen receptor ligands.
These ligands are used as examples only to demonstrate the current
invention of using MDM2 E3 ligase to degrade a target protein and
in no way limit the present invention. In a particular embodiment,
the PTM is selected from the group consisting of:
##STR00095##
wherein "*" indicates one of the positions to connect linker.
[0356] The following is an example of EZH2 ligand. The ligands is
used as an example only to demonstrate the current invention of
using MDM2 E3 ligase to degrade a target protein and in no way
limit the present invention. In a particular embodiment, the PTM is
selected from the group consisting of:
##STR00096##
wherein "*" indicates one of the positions to connect linker.
[0357] The following is an example of JNK ligand. The ligands is
used as an example only to demonstrate the current invention of
using MDM2 E3 ligase to degrade a target protein and in no way
limit the present invention. In a particular embodiment, the PTM is
selected from the group consisting of:
##STR00097##
wherein "*" indicates one of the positions to connect linker.
[0358] The following are examples of MDM2 ligand derived chimeric
molecules using MDM2 E3 ligase to degrade BRDs and androgen
receptor and in no way limit the present invention. In an
embodiment, the chimeric molecule is selected from the group
consisting of:
##STR00098## ##STR00099## ##STR00100## ##STR00101##
[0359] The following are examples of MDM2 ligand derived chimeric
molecules using VHL E3 ligase to degrade MDM2, which provides
examples for Formula (B) as described herein. Thus, in certain
embodiments, the description provides a bifunctional molecules
selected from the group consisting of:
##STR00102## ##STR00103## ##STR00104##
[0360] The following are examples of MDM2 ligand derived chimeric
molecules using MDM2 E3 ligase to degrade androgen receptor. In an
embodiment, the chimeric molecule is selected from the group
consisting of:
##STR00105## ##STR00106## ##STR00107##
[0361] The following are examples of MDM2 ligand derived chimeric
molecules using MDM2 E3 ligase to degrade EZH2. Thus, in certain
additional embodiments, the description provides a bifunctional
compound selected from the group consisting of:
##STR00108##
[0362] The following are examples of MDM2 ligand derived chimeric
molecules using MDM2 E3 ligase to degrade JNK. Thus, in certain
additional embodiments, the description provides a bifunctional
compound selected from the group consisting of:
##STR00109## ##STR00110##
[0363] In certain aspects, the description provides a composition,
e.g., a pharmaceutical composition or therapeutic composition
comprising an effective amount of at least one compound as
described or exemplified herein, and a pharmaceutically acceptable
excipient.
[0364] Synthetic Procedures
[0365] Compounds claimed in this document can be synthesized using
synthetic methods known in the art of organic chemistry. The
following examples are representatives of claimed compounds. All
MDM2 ligand-derived PROTACs disclosed in this document were
analyzed for purity by LC/MS and all final compounds had purity
larger than 95%.
Example 1
Synthesis of A1876
##STR00111##
[0366] Step 1: Synthesis of tert-butyl
N-(14-[2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetra-
azatricyclo[8.3.0.0
[2,6]]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido]-3,6,9,12-tetraoxatet-
radecan-1-yl)carbamate
[0367] Into a 100-mL round-bottom flask, was placed
2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetraazatric-
yclo[8.3.0.0 [2,6]]trideca-2(6),4,7,10,12-pentaen-9-yl]acetic acid
(70.0 mg, 0.17 mmol, 1.00 equiv, prepared from the corresponding
carboxylic acid tert-butyl ester as described in the literature:
Chem. & Bio. 2015, 22, 755-763; PNAS 2016, June 6), tert-butyl
N-(14-amino-3,6,9,12-tetraoxatetradecan-1-yl)carbamate (59.0 mg,
0.18 mmol, 1.00 equiv),
O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium
hexafluorophosphate (80.0 mg, 0.21 mmol, 1.20 equiv),
N,N-Diisopropylethylamine (0.1 mL, 3.00 equiv),
N,N-dimethylformamide (1.0 mL). The resulting solution was stirred
for 1.0 h at room temperature and diluted with water (20 mL). The
mixture was extracted with ethyl acetate (3.times.10 mL) and the
organic layers were combined. The residue was applied onto a silica
gel column and eluted with dichloromethane/methanol (10/1). This
resulted in 160 mg (crude) of tert-butyl
N-(14-[2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetra-
azatricyclo[8.3.0.0
[2,6]]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido]-3,6,9,12-tetraoxatet-
radecan-1-yl)carbamate as a colorless oil.
[0368] Step 2: Synthesis of
N-(14-amino-3,6,9,12-tetraoxatetradecan-1-yl)-2-[(9S)-7-(4-chlorophenyl)--
4,5,13-trimethyl-3-thia-1,8,11,12-tetraazatricyclo[8.3.0.0
[2,6]]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamide Into a 100-mL
round-bottom flask purged and maintained with an inert atmosphere
of nitrogen, was placed tert-butyl
N-(14-[2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetra-
azatricyclo[8.3.0.0
[2,6]]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido]-3,6,9,12-tetraoxatet-
radecan-1-yl)carbamate (160 mg, 0.22 mmol, 1.00 equiv), hydrogen
chloride/dioxane (10.0 mL). The resulting solution was stirred for
1 h at room temperature. The resulting mixture was concentrated
under vacuum. This resulted in 150.0 mg (crude) of
N-(14-amino-3,6,9,12-tetraoxatetradecan-1-yl)-2-[(9S)-7-(4-chlorophenyl)--
4,5,13-trimethyl-3-thia-1,8,11,12-tetraazatricyclo[8.3.0.0
[2,6]]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamide as a white
solid.
Step 3:
3-(3-Chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-{4-[(14--
{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetraazatric-
yclo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}-3,6,9,-
12-tetraoxatetradecan-1-yl)carbamoyl]-2-methoxyphenyl}-4-cyano-5-(2,2-dime-
thylpropyl)pyrrolidine-2-carboxamide (A1876)
[0369] Into a 100-mL round-bottom flask purged and maintained with
an inert atmosphere of nitrogen, was placed
N-(14-amino-3,6,9,12-tetraoxatetradecan-1-yl)-2-[(9S)-7-(4-chlorophenyl)--
4,5,13-trimethyl-3-thia-1,8,11,12-tetraazatricyclo[8.3.0.0
[2,6]]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamide (93.0 mg, 0.15
mmol, 1.00 equiv), racemate of
4-[(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)--
4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-amido]-3-methoxybenzoic
acid (80.0 mg, 0.13 mmol, 1.00 equiv, prepared according to
literature procedure, J. Med. Chem. 2013, 56, 5979),
O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium
hexafluorophosphate (60.0 mg, 0.16 mmol, 1.20 equiv),
N,N-Diisopropylethylamine (0.5 mL, 3.00 equiv),
N,N-dimethylformamide (3.0 mL). The resulting solution was stirred
for 1.0 h at room temperature and diluted with water (20 mL). The
mixture was extracted with ethyl acetate (3.times.10 mL) and the
organic layers were combined. The crude product was purified by
Prep-HPLC (mobile phase:water (10 mmol/L NH.sub.4HCO.sub.3) and
CH.sub.3CN, 6 min, UV detector at 220 nm), which resulted in 10.7
mg (6%) of A1876 as a white solid.
[0370] .sup.1H NMR (300 MHz, CD.sub.3OD): .delta.8.35-8.20 (m, 1H),
7.80-7.70 (m, 1H), 7.55-7.30 (m, 9H), 7.29-7.12 (m, 2H), 4.80-4.70
(m, 1H), 4.68-4.58 (m, 2H), 4.18-3.99 (m, 1H), 3.88 (s, 3H),
3.71-3.54 (m, 18H), 3.52-3.40 (m, 3H), 2.73 (s, 3H), 2.45 (s, 3H),
1.62-1.50 (m, 4H), 1.30-1.25 (m, 2H), 0.98 (s, 9H); LC-MS
calculated for C.sub.60H.sub.66C.sub.13F.sub.2N.sub.9O.sub.8S (m/z)
1216.38, obsd 1216, 1218 [MH.sup.+, Cl.sup.35 and Cl.sup.37],
t.sub.R=2.49 (3.5 minute run).
Example 2
Preparation of A1893 and A1894
##STR00112##
[0371] Preparation of (2R*,3
S*,4R*,5S*)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-{4--
[(14-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetraaz-
atricyclo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}-3-
,6,9,12-tetraoxatetradecan-1-yl)carbamoyl]-2-methoxyphenyl}-4-cyano-5-(2,2-
-dimethylpropyl)pyrrolidine-2-carboxamide (A1893) and
(2S*,3R*,4S*,5R*)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-
-N-{4-[(14-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-t-
etraazatricyclo[8.3.0.0.sup.2,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetam-
ido}-3,6,9,12-tetraoxatetradecan-1-yl)carbamoyl]-2-methoxyphenyl}-4-cyano--
5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (A1894)
[0372] A1876 was separated by preparative LC with a chiral column
to provide two fractions as A1893 and A1894.
[0373] Fraction 1: .sup.1H NMR (300 MHz, CD.sub.3OD):
.delta.8.35-8.27 (m, 1H), 7.80-7.70 (m, 1H), 7.55-7.30 (m, 9H),
7.29-7.12 (m, 2H), 4.80-4.70 (m, 1H), 4.68-4.58 (m, 2H), 4.18-3.99
(m, 1H), 3.88 (s, 3H), 3.71-3.54 (m, 18H), 3.52-3.40 (m, 3H),
3.30-3.25 (m, 1H), 2.73 (s, 3H), 2.45 (s, 3H), 1.62-1.50 (m, 4H),
1.30-1.25 (m, 1H), 0.98 (s, 9H); LC- calculated for
C.sub.60H.sub.66C.sub.13F.sub.2N.sub.9O.sub.8S (m/z) 1216.38, obsd
1216, 1218 [MH.sup.+, Cl.sup.35 and Cl.sup.37], t.sub.R=1.96 min
(2.9 minute run).
[0374] Fraction 2: .sup.1H NMR (300 MHz, CD.sub.3OD):
.delta.8.35-8.27 (m, 1H), 7.80-7.70 (m, 1H), 7.55-7.30 (m, 9H),
7.29-7.12 (m, 2H), 4.80-4.70 (m, 1H), 4.68-4.58 (m, 2H), 4.18-3.99
(m, 1H), 3.88 (s, 3H), 3.71-3.54 (m, 18H), 3.52-3.40 (m, 3H),
2.73-2.60 (m, 4H), 2.45 (s, 3H), 1.62-1.50 (m, 4H), 1.30-1.25 (m,
1H), 0.98 (s, 9H); LC-MS calculated for
C.sub.60H.sub.66C.sub.13F.sub.2N.sub.9O.sub.8S (m/z) 1216.38, obsd
1216, 1218 [MH.sup.+, Cl.sup.35 and Cl.sup.37], t.sub.R=1.96 min
(2.9 minute run).
[0375] Using the same synthetic method as described for A1876,
A1893 and A1894, the following molecules were prepared: A1283,
A1306, A1307, A1863, A1864, A1865, A1829, A1874, A1875, A1890,
A1891 and a1892. The MDM2 ligand for A1283, A1306 and A1306 was
synthesized according to literature procedure (J. Med. Chem. 2013,
56, 5979)
Example 3
Preparation of A1895
##STR00113## ##STR00114## ##STR00115##
[0376] Step 1: Synthesis of
4-(4-methyl-1,3-thiazol-5-yl)benzonitrile
[0377] To a stirred solution of 4-bromobenzonitrile (20 g, 109.88
mmol) in DMA (250 mL) under a nitrogen atmosphere was added
4-methyl-1,3-thiazole (21.88 g, 220.67 mmol), palladium (II)
acetate (743 mg, 3.31 mmol) and potassium acetate (21.66 g, 220.71
mmol) at rt. The resulting solution was heated to 150.degree. C.
and stirred at this temperature for 5 h, at which time LC-MS
indicated completion of reaction. The reaction was cooled to rt,
diluted with 1 L of water and extracted with ethyl acetate (300
mL.times.3). The organic layers were combined, washed with
saturated aqueous solution of sodium chloride (200 mL), dried over
anhydrous sodium sulfate and then concentrated under reduced
pressure to give a crude residue, which was purified by flash
silica gel chromatography (eluent: ethyl acetate/petroleum ether,
v:v=1:5) to give the titled compound (yield: 91%) as a white
solid.
Step 2: Synthesis of
[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methanamine
[0378] To a stirred solution of
4-(4-methyl-1,3-thiazol-5-yl)benzonitrile (35 g, 174.77 mmol) in
tetrahydrofuran (1000 mL) was added LiAlH.sub.4 (20 g, 526.32 mmol)
in portions at 0.degree. C. in 10 min under a nitrogen atmosphere.
The resulting solution was then stirred at 60.degree. C. for 3 h,
at which time LC-MS indicated completion of reaction. The reaction
was cooled to 0.degree. C., then quenched by the addition water (20
mL, added slowly), aq. solution of NaOH (15%, 20 mL) and water (60
mL). The resulting mixture was then extracted with ethyl acetate
(300 mL.times.2). The organic layers were combined, washed with
saturated aqueous solution of sodium chloride (100 mL), dried over
anhydrous sodium sulfate and then concentrated under reduced
pressure to give a crude residue, which was purified by flash
silica gel chromatography (eluent: dichloromethane/methanol
(v:v=10:1)) to give the titled compound (yield: 56%) as a yellow
oil.
Step 3: Synthesis of tert-butyl
(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamo-
yl)pyrrolidine-1-carboxylate
[0379] To a stirred solution of
(2S,4R)-1-[(tert-butoxy)carbonyl]-4-hydroxypyrrolidine-2-carboxylic
acid (2.7 g, 11.68 mmol) in N,N-dimethylformamide (20 mL) was added
DIEA (2.52 g, 19.50 mmol), HATU (4.47 g, 11.76 mmol) and
[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methanamine (2 g, 9.79 mmol)
at rt. The resulting mixture was stirred at rt overnight, at which
time LC-MS indicated completion of reaction. The reaction mixture
was diluted by 20 mL of water and extracted with ethyl acetate (50
mL.times.3). The organic layers were combined, washed with
saturated aqueous solution of sodium chloride (50 mL), dried over
anhydrous sodium sulfate and then concentrated under reduced
pressure to give a crude residue, which was purified by flash
silica gel chromatography (eluent: dichloromethane/methanol
(v:v=20:1)) to give the titled compound (yield: 56%) as a yellow
solid.
Step 4: Synthesis of
(2S,4R)-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolid-
ine-2-carboxamide hydrochloride
[0380] To a solution of tert-butyl
(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamo-
yl)pyrrolidine-1-carboxylate (45 g, 107.78 mmol) in dioxane was
added a solution of hydrogen chloride (13.44 L) in dioxane (300
mL). The resulting solution was stirred for 2 h at 20.degree. C.
The solids were collected by filtration to give the titled product
(yield: 98%) as a yellow solid.
Step 5: Synthesis of tert-butyl
N-[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]met-
hyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbama
[0381] To a stirred solution of
(2S)-2-[[(tert-butoxy)carbonyl]amino]-3,3-dimethylbutanoic acid
(15.73 g, 68.01 mmol) in N,N-dimethylformamide (500 mL) was added
DIEA (29.2 g, 225.94 mmol), HATU (25.9 g, 68.12 mmol, 1.20 equiv)
and
(2S,4R)-2-amino-5-chloro-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)pheny-
l]methyl]pentanamide (20 g, 56.52 mmol) at rt. The resulting
solution was stirred at rt for 16 h, at which time LC-MS indicated
completion of reaction. The reaction mixture was diluted by water
(200 mL) and extracted with ethyl acetate (200 mL.times.3). The
organic layers were combined, washed with saturated aqueous
solution of sodium chloride (50 mL.times.2), dried over anhydrous
sodium sulfate and then concentrated under reduced pressure to give
a crude residue, which was purified by flash silica gel
chromatography (eluent: ethyl acetate/petroleum ether (v:v=2:1)) to
give the titled compound (yield: 51%) as a yellow solid.
Step 6: Synthesis of (2S,
4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-t-
hiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide
hydrochloride
[0382] To a stirred solution of tert-butyl
N-[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]met-
hyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate
(12 g, 22.61 mmol) in dioxane (20 mL) was added a solution of
hydrogen chloride (3.584 L) in dioxane (80 mL) at rt. The resulting
solution was stirred at rt for 2 h, at which time LC-MS indicated
completion of reaction. Precipitated solids were collected by
filtration to give the titled product (yield: 48%) as a yellow
solid. .sup.1HNMR (400 MHz, CD.sub.3OD): .delta. 9.84-9.82 (s, 1H),
7.58-7.54 (m, 4H), 4.71-4.41 (m, 4H), 4.13-4.08 (m, 1H), 3.86-3.71
(m, 2H), 3.36 (s, 1H), 2.60-2.58 (s, 3H), 2.35-2.07 (m, 2H),
1.19-1.12 (m, 9H). LC-MS (ES.sup.+): m/z 431.11 [MH.sup.+],
t.sub.R=0.73 min.
Step 7: Synthesis of tert-butyl
N-(2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethyl)carbamate
[0383] Into a 100-mL round-bottom flask, was placed a solution of
2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethan-1-ol (3.0 g, 15.52 mmol,
1.00 equiv) in tetrahydrofuran/water (30/30 mL), di-tert-butyl
dicarbonate (3.6 g, 16.49 mmol, 1.05 equiv), sodium hydroxide (2.5
g, 62.50 mmol, 4.00 equiv). The resulting solution was stirred for
16 h at room temperature. The resulting solution was diluted with
water (20 mL) and extracted with ethyl acetate (20 mL.times.3). The
combined organic layers were washed with brine (20 mL.times.1),
dried over anhydrous sodium sulfate and concentrated under vacuum.
The residue was applied onto a silica gel column and eluted with
ethyl acetate/petroleum ether (1:1). This resulted in 2.0 g (44%)
of tert-butyl
N-(2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethyl)carbamate as light
yellow oil. LC-MS (ES.sup.+): m/z 294.05 [MH.sup.+], t.sub.R=0.93
min, (1.9 minute run).
Step 8: Synthesis of
14-[[(tert-butoxy)carbonyl]amino]-3,6,9,12-tetraoxatetradecanoic
acid
[0384] Into a 50-mL round-bottom flask, was placed a solution of
tert-butyl
N-(2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethyl)carbamate (300.0
mg, 1.02 mmol, 1.00 equiv) in tetrahydrofuran (10 mL), sodium
hydride (50.0 mg, 2.08 mmol, 1.20 equiv), 2-bromoacetic acid (141.0
mg, 1.01 mmol, 1.00 equiv). The resulting solution was stirred for
4 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). The combined organic layers
were washed with brine (20 mL.times.1). The resulting mixture was
concentrated under vacuum. This resulted in 310.0 mg (86%) of
14-[[(tert-butoxy)carbonyl]amino]-3,6,9,12-tetraoxatetradecanoic
acid as light yellow oil.
Step 9: Synthesis of tert-butyl
N-(1-[[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl-
]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]-
-2,5,8,11-tetraoxatridecan-13-yl)carbamate
[0385] Into a 25-mL round-bottom flask, was placed a solution of
14-[[(tert-butoxy)carbonyl]amino]-3,6,9,12-tetraoxatetradecanoic
acid (175.0 mg, 0.50 mmol, 1.00 equiv) in N,N-dimethylformamide (10
mL),
(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthia-
zol-5-yl)benzyl) pyrrolidine-2-carboxamide hydrochloride (250.0 mg,
0.54 mmol, 1.10 equiv, from Step 6),
O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium
hexafluorophosphate (277.0 mg, 0.73 mmol, 1.20 equiv),
N,N-diisopropylethylamine (0.5 mL, 5.00 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). The
combined organic layers were washed with brine (20 mL.times.1). 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 227.0
mg (60%) of tert-butyl
N-(1-[[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl-
]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]-
-2,5,8,11-tetraoxatridecan-13-yl)carbamate as yellow oil. LC-MS
(ES.sup.+): m/z 764.35 [MH.sup.+], t.sub.R=1.08 min, (1.9 minute
run).
Step 10: Synthesis of
(2S,4R)-1-[(2S)-2-(14-amino-3,6,9,12-tetraoxatetradecanamido)-3,3-dimethy-
lbutanoyl]-4-hydroxy-N-[
[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide
[0386] Into a 25-mL round-bottom flask, was placed a solution of
tert-butyl
N-(1-[[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl-
]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]-
-2,5,8,11-tetraoxatridecan-13-yl)carbamate (227.0 mg, 0.30 mmol,
1.00 equiv) in dioxane (10 mL), then HCl (g) was introduced in. The
resulting solution was stirred for 2 h at room temperature. The
resulting mixture was concentrated under vacuum. This resulted in
190.0 mg (96%) of
(2S,4R)-1-[(2S)-2-(14-amino-3,6,9,12-tetraoxatetradecanamido)-3,3-dimethy-
lbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrol-
idine-2-carboxamide as yellow oil.
Step 11: Synthesis of
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-[4-[(1-[[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-th-
iazol-5-yl)phenyl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobuta-
n-2-yl]carbamoyl]-2,5,8,11-tetraoxatridecan-13-yl)carbamoyl]-2-methoxyphen-
yl]pyrrolidine-2-carboxamide (A1895)
[0387] Into a 50-mL round-bottom flask, was placed a solution of
racemate of 4-[(2R, 3S, 4R,
5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,-
2-dimethylpropyl)pyrrolidine-2-amido]-3-methoxybenzoic acid (80 mg,
0.13 mmol, 1.00 equiv) in N,N-dimethylformamide (10 mL),
(2S,4R)-1-[(2S)-2-(14-amino-3,6,9,12-tetraoxatetradecanamido)-3,3-dimethy-
lbutanoyl]-4-hydroxy-N-[ [4-(4-methyl-1,3-thiazol-5-yl)phen
yl]methyl]pyrrolidine-2-carboxamide (320.0 mg, 0.48 mmol, 1.10
equiv), O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium
hexafluorophosphate (64.0 mg, 0.17 mmol, 1.20 equiv),
N,N-diisopropylethylamine (0.4 mL, 5.00 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
combined organic layers were washed with brine (20 mL.times.1). The
mixture was dried over anhydrous sodium sulfate and concentrated
under vacuum. The crude material was purified by prep-HPLC (column:
XBridge Shield RP18 OBD Column, Sum, 19*150 mm; Mobile Phase A:
water (10 mmol/L bicarbonate amine), Mobile Phase B: acetonitrile;
Flow rate: 20 mL/min; Gradient: 50% B to 60% B in 9 min; 254 nm).
This resulted in 40.0 mg (24%) of (2R/2S, 3S/3R, 4R/4S,
5S/SR)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5--
(2,2-dimethylpropyl)-N-[4-[(1-[[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-
-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1--
oxobutan-2-yl]carbamoyl]-2,5,8,11-tetraoxatridecan-13-yl)carbamoyl]-2-meth-
oxyphenyl]pyrrolidine-2-carboxamide as a white solid.
[0388] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.84 (s, 1H),
8.34-8.31 (m, 1H), 7.62-7.64 (m, 1H), 7.51 (s, 1H), 7.40-7.29 (m,
8H), 7.24-7.19 (m, 2H), 4.80-4.75 (m, 1H), 4.70-4.55 (m, 3H),
4.55-4.45 (m, 1H), 4.27-4.25 (m, 1H), 4.10-4.00 (m, 1H), 3.99-3.77
(m, 5H), 3.70-3.66 (m, 2H), 3.66-3.61 (m, 15H), 3.59-3.55 (m, 2H),
2.48 (s, 3H), 2.25-2.19 (m, 1H), 2.19-1.92 (m, 1H), 1.66-1.63 (m,
1H), 1.40-1.35 (m, 1H), 0.99 (s, 18H). LC-MS calcd for
C.sub.63H.sub.76Cl.sub.2F.sub.2N.sub.8O.sub.11S (m/z) 1260.47, obsd
1284.05/1286.05 [M+Na.sup.+], t.sub.R=2.27 min, (3.6 minute
run).
Example 4
Preparation of A1896 and a1897
##STR00116##
[0389] Preparation of (2R*,3
S*,4R*,5S*)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cya-
no-5-(2,2-dimethylpropyl)-N-{4-[(1-{[(2S)-1-[(2S,4R)-4-hydroxy-2-({[4-(4-m-
ethyl-1,3-thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3,3-dimeth-
yl-1-oxobutan-2-yl]carbamoyl}-2,5,8,11-tetraoxatridecan-13-yl)carbamoyl]-2-
-methoxyphenyl}pyrrolidine-2-carboxamide (A1896) and
(2S*,3R*,4S*,5R*)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-
-4-cyano-5-(2,2-dimethylpropyl)-N-{4-[(1-{[(2S)-1-[(2S,4R)-4-hydroxy-2-({[-
4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-1-yl]-3,3--
dimethyl-1-oxobutan-2-yl]carbamoyl}-2,5,8,11-tetraoxatridecan-13-yl)carbam-
oyl]-2-methoxyphenyl}pyrrolidine-2-carboxamide (A1897)
[0390] A1895 was separated by Prep-chiral-HPLC (column: Phenomenex
Lux 5u Cellulose-4, AXIA Packed 250*21.2 mm, Sum; Mobile Phase:
methanol in water, Flow rate: 20 mL/min; run time: 24 min; 254/220
nm). Two fractions were collected. Fraction A (RT1: 11.68 min) gave
A1896 (15 mg) as a white solid.
[0391] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.83 (s, 1H),
8.34-8.31 (m, 1H), 7.62-7.64 (m, 1H), 7.51 (s, 1H), 7.40-7.29 (m,
8H), 7.24-7.19 (m, 2H), 4.80-4.74 (m, 3H), 4.70-4.55 (m, 3H),
4.33-4.28 (m, 1H), 4.06-4.03 (m, 1H), 3.98-3.94 (m, 5H), 3.81-3.77
(m, 2H), 3.66-3.61 (m, 16H), 2.48 (s, 3H), 2.25-2.19 (m, 1H),
2.19-1.92 (m, 1H), 1.66-1.63 (m, 1H), 1.40-1.35 (m, 1H), 0.99 (s,
18H). LC-MS calcd for
C.sub.63H.sub.76Cl.sub.2F.sub.2N.sub.8O.sub.11S (m/z) 1260.47, obsd
1284.05/1286.05 [M+Na.sup.+], t.sub.R=2.27 min, (3.6 minute
run).
[0392] Fraction B (RT2: 20.22 min) gave A1897 also as a white solid
(15 mg).
[0393] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.83 (s, 1H),
8.34-8.31 (m, 1H), 7.62-7.64 (m, 1H), 7.51 (s, 1H), 7.40-7.29 (m,
8H), 7.24-7.19 (m, 2H), 4.80-4.55 (m, 6H), 4.33-4.28 (m, 1H),
4.06-4.03 (m, 1H), 3.98-3.94 (m, 5H), 3.81-3.77 (m, 2H), 3.66-3.61
(m, 16H), 2.48 (s, 3H), 2.25-2.19 (m, 1H), 2.19-1.92 (m, 1H),
1.72-1.63 (m, 1H), 1.40-1.25 (m, 1H), 1.02 (s, 18H). LC-MS calcd
for C.sub.63H.sub.76Cl.sub.2F.sub.2N.sub.8O.sub.11S (m/z) 1260.47,
obsd 1284.05/1286.05 [M+Na.sup.+], t.sub.R=2.42 min, (3.6 minute
run).
[0394] Using the same synthetic method as described for A1895,
A1896 and A1897, the following molecules were prepared: A1877,
A1907, A1908, A1909, A1910, and A1911.
Example 5
Preparation of A1717
##STR00117## ##STR00118##
[0395] Step 1: Synthesis of tert-butyl
N-[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]-
carbamate
[0396] Into a 100-mL 3-necked round-bottom flask purged and
maintained with an inert atmosphere of nitrogen, was placed a
solution of tert-butyl
N-[(1,3-trans)-3-hydroxy-2,2,4,4-tetramethylcyclobutyl]carbamate
(600.0 mg, 2.47 mmol, 1.00 equiv) in N,N-dimethylformamide (10.0
mL). This was followed by the addition of sodium hydride (198.0 mg,
8.25 mmol, 2.00 equiv), in portions at 0.degree. C. After 30 min,
to this was added 2-chloro-4-fluorobenzonitrile (459.0 mg, 2.95
mmol, 1.20 equiv). The resulting solution was stirred for 1 h at
70.degree. C. The reaction mixture was cooled to room temperature
with a water bath. 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 were combined. The
resulting solution was washed with brine and concentrated. The
residue was applied onto a silica gel column eluted with ethyl
acetate/petroleum ether (1/5). This resulted in 100.0 mg (11%) of
tert-butylN-[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetrameth-
ylcyclobutyl]carbamate as colorless oil. LC-MS (ES.sup.+): m/z
279.10 [MH-100].sup.+, t.sub.R=1.20 min (2.5 minute run).
Step 2: Synthesis of
2-chloro-4-[(1,3-trans)-3-amino-2,2,4,4-tetramethylcyclobutoxy]-benzonitr-
ile
[0397] Into a 100-mL round-bottom flask purged and maintained with
an inert atmosphere of nitrogen, was placed tert-butyl
N-[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]-
carbamate (500.0 mg, 1.32 mmol, 1.00 equiv), hydrogen
chloride/dioxane (3 mL, 4M), 1,4-dioxane (3 mL). The resulting
solution was stirred for 1 h at room temperature. The resulting
mixture was concentrated under vacuum. This resulted in 447 mg
(87%) of
2-chloro-4-[(1,3-trans)-3-amino-2,2,4,4-tetramethylcyclobutoxy]benzonitri-
le as a white solid.
Step 3: Synthesis of
4-[[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl-
]carbamoyl]phenyl acetate
[0398] Into a 25-mL round-bottom flask, was placed a solution of
4-(acetyloxy)benzoic acid (100.0 mg, 0.56 mmol, 1.00 equiv) in
N,N-dimethylformamide (10 mL),
2-chloro-4-[(1,3-trans)-3-amino-2,2,4,4-tetramethylcyclobutoxy]benzonitri-
le (190.0 mg, 0.68 mmol, 1.10 equiv), HATU (253.0 g, 665.39 mmol,
1.20 equiv), DIEA (0.5 mL, 5.00 equiv). The resulting solution was
stirred for 2 h at room temperature. The reaction was then quenched
by the addition of water (10 mL). The resulting solution was
extracted with ethyl acetate (10 mL.times.3) and the organic layers
were combined. The resulting mixture was washed with brine (10
mL.times.1). The mixture was dried over anhydrous sodium sulfate
and concentrated under vacuum. This resulted in 230.0 mg (94%) of
4-[[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl-
]carbamoyl]phenyl acetate as a light brown solid. LC-MS (ES.sup.+):
m/z 441.00 [M+H.sup.+], t.sub.R=1.09 min
Step 4: Synthesis of
4-hydroxy-N-[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylc-
yclobutyl]benzamide
[0399] Into a 50-mL round-bottom flask, was placed
4-[[(R1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobuty-
l]carbamoyl]phenyl acetate (230.0 mg, 0.52 mmol, 1.00 equiv),
sodium hydroxide (100.0 mg, 2.50 mmol, 3.00 equiv) and methanol (10
mL). The resulting solution was stirred at room temperature. The
resulting mixture was concentrated under vacuum and diluted with
water (10 mL). The pH value of the solution was adjusted to 4-5
with hydrogen chloride (1 mol/L). The resulting solution was
extracted with ethyl acetate (10 mL.times.3) and the organic layers
were combined. The solution was washed with brine (10 mL.times.1)
and concentrated under vacuum. The residue was applied onto a
silica gel column eluted with ethyl acetate/petroleum ether (1:1).
This resulted in 200.0 mg (96%) of
4-hydroxy-N-[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylc-
yclobutyl]-benzamide as light yellow oil.
Step 5: Synthesis of 2-[2-(benzyloxy)ethoxy]ethyl
4-methylbenzene-1-sulfonate
[0400] Into a 250-mL round-bottom flask,
2-[2-(benzyloxy)ethoxy]ethan-1-ol (3.0 g, 15.29 mmol, 1.00 equiv),
4-methylbenzene-1-sulfonyl chloride (4.36 g, 22.87 mmol, 1.50
equiv), triethylamine (3.09 g, 30.54 mmol, 2.00 equiv),
4-dimethylaminopyridine (933.0 mg, 7.64 mmol, 0.50 equiv) were
mixed in dichloromethane (20 mL). The resulting solution was
stirred for 3 h at room temperature. The mixture was applied onto a
silica gel column eluted with ethyl acetate/petroleum ether (1/3).
This resulted in 4.9 g (91%) of 2-[2-(benzyloxy)ethoxy]ethyl
4-methylbenzene-1-sulfonate as light yellow oil.
Step 6: Synthesis of tert-butyl
N-(1-phenyl-2,5,8,11,14-pentaoxahexadecan-16-yl)carbamate
[0401] Into a 25-mL round-bottom flask, sodium hydride (67 mg, 2.79
mmol, 1.20 equiv, 60% in oil) was added to a solution of tert-butyl
N-2-[2-(2-hydroxyethoxy)ethoxy]ethylcarbamate (320.0 mg, 1.28 mmol,
1.00 equiv) in N,N-dimethylformamide (10 mL) at 0.degree. C. The
mixture was stirred for 15 min at that temperature. Then
2-[2-(benzyloxy)ethoxy]ethyl 4-methylbenzene-1-sulfonate (536 mg,
1.53 mmol, 1.20 equiv) was added and the reaction was warmed to
room temperature and stirred for 4 h. After quenched by addition of
water, the resulting solution was extracted with ethyl acetate (50
mL.times.3) and the organic layers were combined. The resulting
mixture was washed with brine (50 mL.times.3) and dried over
anhydrous sodium sulfate. The filtered solution was concentrated
and the residue was applied onto a silica gel column eluted with
ethyl acetate/petroleum ether (4/1). This resulted in 290 mg (53%)
of tert-butyl
N-(1-phenyl-2,5,8,11,14-pentaoxahexadecan-16-yl)carbamate as light
yellow oil. LC-MS (ES.sup.+): m/z 428.95 [M+H.sup.+], t.sub.R=0.92
min, (1.9 minute run).
Step 7: Synthesis of tert-butyl
14-hydroxy-3,6,9,12-tetraoxatetradecylcarbamate
[0402] Into a 50-mL round-bottom flask, palladium on carbon (200.0
mg) was added to a solution of tert-butyl
N-(1-phenyl-2,5,8,11,14-pentaoxahexadecan-16-yl)carbamate (290.0
mg, 0.68 mmol, 1.00 equiv) in methanol (10 mL) at room temperature
under nitrogen atmosphere. The reaction flask was vacuumed and
charged with a hydrogen balloon. The resulting solution was stirred
for 8 h at 40.degree. C. in an oil bath. The reaction mixture was
then filtered through a Celite pad and the filtrate was
concentrated under reduced pressure. This resulted in 220 mg
(crude) of tert-butyl
14-hydroxy-3,6,9,12-tetraoxatetradecylcarbamate as yellow oil
Step 8: Synthesis of tert-butyl
N-(14-[[(4-methylbenzene)sulfonyl]oxy]-3,6,9,12-tetraoxatetradecan-1-yl)c-
arbamate
[0403] Into a 25-mL round-bottom flask, tert-butyl
N-(14-hydroxy-3,6,9,12-tetraoxatetradecan-1-yl)carbamate (228.0 mg,
0.68 mmol, 1.00 equiv), 4-methylbenzene-1-sulfonyl chloride (192.0
mg, 1.01 mmol, 1.50 equiv), triethylamine (136.2 mg, 1.35 mmol,
2.00 equiv), 4-dimethylaminopyridine (16.4 mg, 0.13 mmol, 0.20
equiv) were mixed in dichloromethane (10 mL). The resulting
solution was stirred for 8 h at room temperature. The residue was
applied onto a silica gel column eluted with ethyl
acetate/petroleum ether (3/2). This resulted in 110.0 mg (33%) of
tert-butyl
N-(14-[[(4-methylbenzene)sulfonyl]oxy]-3,6,9,12-tetraoxatetradecan-1-yl)c-
arbamate as light yellow oil. LC-MS (ES.sup.+): m/z 492.00
[M+H.sup.+], t.sub.R=0.93 min, (1.9 minute run).
Step 9: Synthesis of tert-butyl
N-[1-(4-[[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcycl-
obutyl]carbamoyl]phenyl)-1,4,7,10,13-pentaoxapentadecan-15-yl]carbamate
[0404] Into a 25-mL round-bottom flask, was placed
4-hydroxy-N-[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylc-
yclobutyl]benzamide (50.0 mg, 0.13 mmol, 1.00 equiv), potassium
carbonate (34.7 mg, 0.25 mmol, 2.00 equiv), tert-butyl
N-(14-[(4-methylbenzene)sulfonyl]oxy-3,6,9,12-tetraoxatetradecan-1-yl)car-
bamate (74.0 mg, 0.15 mmol, 1.20 equiv) in N,N-dimethylformamide
(5.0 mL). The resulting solution was stirred for 5 h at 80.degree.
C. in an oil bath. The reaction was quenched with 50 mL of water.
The resulting solution was extracted with ethyl acetate (50
mL.times.3) and the organic layers were combined. The resulting
mixture was washed with brine (50 mL.times.3) and dried over
anhydrous sodium sulfate. After the evaporation of solvents, the
crude product was purified by prep-TLC with ethyl acetate/petroleum
ether (4/1). This resulted in 110.0 mg of tert-butyl
N-[1-(4-[[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcycl-
obutyl]carbamoyl]phenyl)-1,4,7,10,13-pentaoxapentadecan-15-yl]carbamate
as light yellow oil. LC-MS (ES.sup.+): m/z 740.10/742.10
[M+Na.sup.+], t.sub.R=1.14 min, (1.9 minute run).
Step 10: Synthesis of
4-[(14-amino-3,6,9,12-tetraoxatetradecan-1-yl)oxy]-N-[(1,3-trans)-3-(3-ch-
loro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide
hydrochloride
[0405] Into a 25-mL round-bottom flask, hydrogen chloride (2 mL, 2N
in dioxane) was added to a solution of tert-butyl N-[1-(4-[
[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]ca-
rbamoyl]phenyl)-1,4,7,10,13-pentaoxapentadecan-15-yl]carbamate
(110.0 mg, 0.15 mmol, 1.00 equiv) in methanol (15 mL). The
resulting solution was stirred for 3 h at room temperature and
concentrated under vacuum. This resulted in 100.0 mg (crude) of
4-[(14-amino-3,6,9,12-tetraoxatetradecan-1-yl)oxy]-N-[(1,3-trans)-3-(3-ch-
loro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide
hydrochloride as light yellow oil.
Step 11: Synthesis of
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-[1-(4-[
[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]ca-
rbamoyl]phenyl)-1,4,7,10,13-pentaoxapentadecan-15-yl]pyrrolidine-2-carboxa-
mide (A1717)
[0406] Into a 25-mL round-bottom flask, was placed
(2R/2S,3S/3R,4R/4S,5S/5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluor-
ophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxylic
acid (59.4 mg, 0.13 mmol, 1.00 equiv, prepared according to
literature procedure: J. Med. Chem. 2013, 56, 5979),
4-[(14-amino-3,6,9,12-tetraoxatetradecan-1-yl)oxy]-N-[(1,3-trans)-3-(3-ch-
loro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide
hydrochloride (100.0 mg, 0.15 mmol, 1.20 equiv),
N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
hexafluorophospate (58.0 mg, 0.15 mmol, 1.20 equiv),
N,N-dimethylformamide (5.0 mL). N,N-Diisopropylethylamine (82.3 mg,
0.64 mmol, 5.00 equiv) was added and the reaction was stirred for 2
h at room temperature. The reaction mixture was quenched by
addition of 20 mL of water. The resulting solution was extracted
with ethyl acetate (50 mL.times.3) and the organic layers were
combined. The resulting mixture was washed with brine (50
mL.times.3) and dried over anhydrous sodium sulfate. After the
evaporation of solvents, the crude product was purified by
Prep-HPLC with the following conditions: Column, XBridge Shield
RP18 OBD Column, 5 um, 19*150 mm; mobile phase:water with 10 mmol/L
ammonium bicarbonate and acetonitrile (hold 74.0% acetonitrile in
10 min); Detector, UV 254 nm. This resulted in 45 mg (33%) of
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-[1-(4-[[(1,3-trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-te-
tramethylcyclobutyl]carbamoyl]phenyl)-1,4,7,10,13-pentaoxapentadecan-15-yl-
]pyrrolidine-2-carboxamide (A1717) as a white solid.
[0407] .sup.1H NMR (400 MHz, CD.sub.3OD): .delta.7.82-7.80 (d,
J=8.0 Hz, 2H), 7.74-7.65 (m, 2H), 7.39-7.31 (m, 3H), 7.26-7.22 (m,
2H), 7.14 (d, J=2.8 Hz, 1H), 7.05-6.98 (m, 3H), 4.73-4.71 (d, J=7.2
Hz, 1H), 4.44-4.42 (d, J=8.4 Hz, 1H), 4.29 (s, 1H), 4.23-4.20 (d,
J=9.2 Hz, 2H), 4.16 (s, 1H), 4.00 (m, 1H), 3.90-3.88 (d, J=9.2 Hz,
2H), 3.74-3.57 (m, 13H), 3.50-3.40 (m, 1H), 3.36-3.32 (m, 1H),
1.65-1.55 (m, 1H), 1.31-1.29 (m, 1H), 1.28 (s, 6H), 1.24 (s, 6H),
0.95 (s, 9H); LC-MS calcd for
C.sub.55H.sub.64Cl.sub.3F.sub.2N.sub.5O.sub.8 (m/z) 1067.48, obsd
1068.50 [M+H.sup.+], t.sub.R=2.76 min, (3.6 minute run).
Example 6
Preparation of A1720 and A1735
##STR00119##
[0408] Preparation of
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-[1-(4-{[(1,3-trans)-3-(3-chloro-4-cyanopheno-
xy)-2,2,4,4-tetramethylcyclobutyl]carbamoyl}phenyl)-1,4,7,10,13-pentaoxape-
ntadecan-15-yl]pyrrolidine-2-carboxamide (A1720) and
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-[1-(4-{[(1,3-trans)-3-(3-chloro-4-cyanopheno-
xy)-2,2,4,4-tetramethylcyclobutyl]carbamoyl}phenyl)-1,4,7,10,13-pentaoxape-
ntadecan-15-yl]pyrrolidine-2-carboxamide (A1735)
[0409] A1717 was separated by preparative chiral HPLC (Column:
Chiralpak IA 2*25 cm, 5 um; Mobile Phase A: hexane; Mobile Phase B:
ethanol; Flow rate: 15 mL/min; Gradient: 50 B to 50 B in 35 min;
254/220 nm). The chiral separation resulted in two fractions.
Fraction A (RT1: 16.962 min) gave 10.0 mg (29%) of
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-[1-(4-[[(1,3-tran)-3-(3-chloro-4-cyanophenox-
y)-2,2,4,4-tetramethylcyclobutyl]carbamoyl]phenyl)-1,4,7,10,13-pentaoxapen-
tadecan-15-yl]pyrrolidine-2-carboxamide as a white solid.
[0410] .sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 7.82-7.80 (d,
J=8.0 Hz, 2H), 7.74-7.65 (m, 2H), 7.39-7.31 (m, 3H), 7.26-7.22 (m,
2H), 7.14 (d, J=2.8 Hz, 1H), 7.05-6.98 (m, 3H), 4.73-4.71 (d, J=7.2
Hz, 1H), 4.44-4.42 (d, J=8.4 Hz, 1H), 4.29 (s, 1H), 4.23-4.20 (d,
J=9.2 Hz, 2H), 4.16 (s, 1H), 4.00 (m, 1H), 3.90-3.88 (d, J=9.2 Hz,
2H), 3.74-3.57 (m, 13H), 3.50-3.40 (m, 1H), 3.36-3.32 (m, 1H),
1.65-1.55 (m, 1H), 1.31-1.29 (m, 1H), 1.28 (s, 6H), 1.24 (s, 6H),
0.95 (s, 9H); LC-MS calcd for
C.sub.55H.sub.64Cl.sub.3F.sub.2N.sub.5O.sub.8 (m/z) 1067.48, obsd
1068.10/1070.10 [M+H.sup.+], t.sub.R=2.62 min, (3.6 minute
run).
[0411] The second fraction (RT2: 28.90 min) gave 10.0 mg (29%) of
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-5-(2,2-dimethylpropyl)-N-[1-(4-[[(1r,3r)-3-(3-chloro-4-cyanophenoxy)--
2,2,4,4-tetramethylcyclobutyl]carbamoyl]phenyl)-1,4,7,10,13-pentaoxapentad-
ecan-15-yl]pyrrolidine-2-carboxamide as a white solid.
[0412] .sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 7.82-7.80 (d,
J=8.0 Hz, 2H), 7.74-7.65 (m, 2H), 7.39-7.31 (m, 3H), 7.26-7.22 (m,
2H), 7.14 (d, J=2.8 Hz, 1H), 7.05-6.98 (m, 3H), 4.73-4.71 (d, J=7.2
Hz, 1H), 4.44-4.42 (d, J=8.4 Hz, 1H), 4.29 (s, 1H), 4.23-4.20 (d,
J=9.2 Hz, 2H), 4.16 (s, 1H), 4.00 (m, 1H), 3.90-3.88 (d, J=9.2 Hz,
2H), 3.74-3.57 (m, 13H), 3.50-3.40 (m, 1H), 3.36-3.32 (m, 1H),
1.65-1.55 (m, 1H), 1.31-1.29 (m, 1H), 1.28 (s, 6H), 1.24 (s, 6H),
0.95 (s, 9H); LC-MS calcd for
C.sub.55H.sub.64Cl.sub.3F.sub.2N.sub.5O.sub.8 (m/z) 1067.48, obsd
1068.10/1070.10 [M+H.sup.+], t.sub.R=2.62 min, (3.6 minute
run).
[0413] Compounds A1751, A1603, A1621 and A1688 were prepared using
the same method as described for the preparation of A1717, A1720
and A1735
Example 7
Preparation of A2434
##STR00120##
[0414] Step 1: Synthesis of
2-chloro-4-isothiocyanatobenzonitrile
[0415] To a stirred solution of 4-amino-2-chlorobenzonitrile (1 g,
6.55 mmol) in dichloromethane (9 mL) was added sodium bicarbonate
(2.21 g, 26.31 mmol) and water (9 mL), followed by addition of
thiophosgene (817 mg, 7.11 mmol) drop wise in 30 min at 0.degree.
C. The resulting mixture was stirred for 1 h at room temperature.
The reaction mixture was diluted with dichloromethane (200 mL),
washed with brine (50 mL.times.2), dried over anhydrous sodium
sulfate and then concentrated under reduced pressure to give a
crude residue. The residue was purified by flash silica gel
chromatography (eluent: ethyl acetate/petroleum ether (v:v=1:30))
to give the desired product (yield: 71%).sup.1HNMR (400 MHz,
CDCl.sub.3): .delta. 7.69 (d, J=8.0 Hz, 1H), 7.38 (s, 1H), 7.28 (m,
1H).
Step 2: Synthesis of 2-chloro-4-[3-(4-hydroxyphenyl)-5-imino-4,
4-dimethyl-2-sulfanylideneimidazolidin-1-yl]benzonitrile
[0416] To a stirred solution of
2-chloro-4-isothiocyanatobenzonitrile (399 mg, 2.05 mmol) in
toluene (5 mL) was added
2-[(4-hydroxyphenyl)amino]-2-methylpropanenitrile (300 mg, 1.70
mmol) and 4-dimethylaminopyridine (312 mg, 2.55 mmol). The
resulting solution was then heated in an oil bath to 100.degree. C.
and stirred at the same temperature for 16 h. The resulting mixture
was concentrated under vacuum. The residue was purified by flash
silica gel chromatography (eluent: ethyl acetate/petroleum ether,
v:v=1:1) to give the desired product (yield: 48%) as a brown solid.
LC-MS (ES.sup.+): m/z 370.95 [M+H.sup.+], t.sub.R=0.74 min.
Step 3: Synthesis of
2-chloro-4-[3-(4-hydroxyphenyl)-4,4-dimethyl-5-oxo-2-sulfanylideneimidazo-
lidin-1-yl]benzonitrile
[0417] To a stirred solution of
2-chloro-4-[3-(4-hydroxyphenyl)-5-imino-4,
4-dimethyl-2-sulfanylideneimidazolidin-1-yl]benzonitrile (300 mg,
0.81 mmol) in methanol (6 mL) was added aqueous hydrogen chloride
(2N, 3.0 mL). The resulting solution was then heated in an oil bath
to 100.degree. C. and stirred at the same temperature for 2 h. The
reaction mixture was diluted with water (30 mL), extracted with
ethyl acetate (60 mL.times.3), washed with water (50 mL), dried
over anhydrous sodium sulfate and concentrated under vacuum to give
titled product (yield: 93%) as a yellow solid, which was used for
next step without any further purifications. LC-MS (ES.sup.+): m/z
372.00 [M+H.sup.+], t.sub.R=0.97 min.
Step 4 and Step 5: Preparation of
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-c-
yano-N-(3-{[5-(4-{3-[4-cyano-3-(trifluoromethyl)phenyl]-5,5-dimethyl-4-oxo-
-2-sulfanylideneimidazolidin-1-yl}phenoxy)pentyl]oxy}propyl)-5-(2,2-dimeth-
ylpropyl)pyrrolidine-2-carboxamide (A2434)
[0418] Step 4 and step 5 were carried out using the method as
described for the synthesis of A1717.
[0419] Compound A2434 was isolated as a solid. LC-MS calcd for
C.sub.50H.sub.51Cl.sub.2F.sub.5N.sub.6O.sub.4S (m/z) 997.94, obsd
997/999.
[0420] Compound A2435 was prepared with the same method as
described for the preparation of A2434.
[0421] In the case of the preparation of compound A679, A680 and
A702, the similar synthetic route was used. The MDM2 ligand of
imidazoline chemotype was synthesized according to literature
procedure (ACS Med. Chem. Lett. 2103, 4, 466).
Example 8
Preparation of A2844
##STR00121## ##STR00122##
[0422] Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic
acid
[0423] To a stirred solution of 2-methyl-3-nitrobenzoic acid (10 g,
55 mmol) in conc. H.sub.2SO.sub.4 (40 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (9 g, 32 mmol) was
added portion wise at room temperature and reaction was stirred at
room temperature for 5 h. Then the reaction mass was poured on an
ice cold water. Solid was filtered, and the resulting residue was
washed with water and dried under vacuum to afford
5-bromo-2-methyl-3-nitrobenzoic acid (12 g, 84%) as a light yellow
solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.28 (d, J=2.0
Hz, 1H), 8.13 (d, J=2.0 Hz, 1H), 2.51 (s, 3H)
Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate
[0424] A mixture of 5-bromo-2-methyl-3-nitrobenzoic acid (12 g, 41
mmol) in SOCl.sub.2/MeOH (v:v=1:10) (250 mL) was heated to reflux
overnight. The reaction mixture was cooled and concentrated. The
residue was dissolved in 300 mL of ethyl acetate. The organic layer
was washed sequentially with sat. aq. NaHCO.sub.3 and brine, dried
over Na.sub.2SO.sub.4, and concentrated. The residue was purified
by chromatography (silica gel, petroleum ether/ethyl acetate (20:1,
v:v)) to afford the desired compound (11 g, yield: 87%). .sup.1H
NMR (400 MHz, CDCl.sub.3): .delta. 8.12 (d, J=2.0 Hz, 1H), 7.97 (d,
J=2.0 Hz, 1H), 3.95 (s, 3H), 2.57 (s, 3H)
Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate
[0425] To a stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzoate (11 g, 40 mmol) in ethanol (100
mL), was added NH.sub.4Cl solution (13 g in 50 mL water, 240 mmol)
followed by Fe powder (20 g, 360 mmol). The resulting reaction was
stirred at 80.degree. C. for 2-3 h. Then the reaction mixture was
filtered and the filtrate was concentrated till dryness to give a
solid which was dissolved in sat. sodium bicarbonate solution.
Aqueous layer was extracted with ethyl acetate (3.times.100 mL).
The combined organic layers were dried over sodium sulfate and
concentrated to afford the desired compound (8.1 g, 83%).
[0426] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.33 (s, 1H),
6.94 (s, 1H), 3.87 (s, 3H), 3.79 (br, 2H), 2.28 (s, 3H)
Step 4: Synthesis of methyl
5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate
[0427] To a solution of methyl 3-amino-5-bromo-2-methylbenzoate (2
g, 8.2 mmol) in dichloromethane (20 mL) and acetic acid (2.5 g, 40
mol) was added tetrahydropyran-4-one (1.2 g, mol 12 mmol) at
25.degree. C. After 2.5 h, NaCNBH.sub.3 was added into the reaction
in portions and the mixture was stirred overnight. The reaction was
quenched with a solution of sodium hydroxide (1.6 g, 40 mmol) in
water (50 mL). After stirring for 10 minutes at ambient
temperature, the organic layer was washed with water (2.times.50
mL), dried (Na.sub.2SO.sub.4) and concentrated. The crude product
was purified by silica gel chromatography eluting with 5-20% ethyl
acetate in petroleum to afford the desired compound (1.3 g, 50%) as
a light yellow oil. .sup.1H NMR 400 MHz, DMSO-d.sub.6): .delta.
6.97 (s, 1H), 6.93 (s, 1H), 4.99 (d, J=8.0 Hz, 1H), 3.87 (d, d,
J=10.80 Hz, 2H), 3.80 (s, 3H), 3.60 (br, 1H), 3.44 (t, J=11.6 Hz,
3H), 2.15 (s, 3H), 1.84 (d, J=12.4 Hz, 2H), 1348-1.57 (m, 2H)
Step 5: Synthesis of methyl
5-bromo-3-[ethyl(oxan-4-yl)amino]-2-methylbenzoate
[0428] To a stirred solution of methyl
5-bromo-2-methyl-3-[(oxan-4-yl)amino]benzoate (1 g, 119 mmol) in
THF (20 mL) was added LiHDMS (1.0M, 2.0 eq, THF) at 0.degree. C.
After 30 min, EtI (4.0 eq) was added into the mixture at 0.degree.
C. Then reaction mixture was stirred at rt for 3 h. Saturated
NaHCO.sub.3 was added and the mixture was separated. The aqueous
layer was extracted with CH.sub.2Cl.sub.2 and the combined organic
layers were concentrated in vacuo to afford the desired product
(1.2 g crude) which was used into next step without further
purification.
Step 6: Synthesis of
5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoic
acid
[0429] To a stirred solution of
5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate
(1.2 g, crude) in ethanol (15 mL) was added LiOH (0.3 g, 10 mmol)
and the resulting mixture was stirred at 60.degree. C. for 1 h.
Upon the completion of the reaction as determined by TLC, the
solvent was removed under reduced pressure and the residue was
acidified with 1N HCl until pH.about.5, and it was concentrated.
The crude product was purified by silica gel chromatography eluting
with 5-10% (CH.sub.3OH/DCM) to afford the desired product (0.7 g,
70%) as a light yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 7.88 (s, 1H), 7.42 (s, 1H), 3.98 (d, J=11.2 Hz, 2H), 3.34
(t, J=11.2 Hz, 2H), 3.03-3.09 (m, 2H), 2.95-3.00 (m, 1H), 2.52 (s,
3H), 1.64-1.73 (m, 4H), 0.88 (t, J=6.8 Hz, 3H)
Step 7: Synthesis of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(t-
etrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide
[0430] The acid from step 6 (0.5 g, 1.5 mmol) was dissolved in DMF
(5 mL), and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.45 g,
2.9 mmol) and DIEA (0.84 g, 5.8 mmol) were added. The reaction
mixture was stirred at room temperature for 15 minutes, and then
PYBOP (1.6 g, 3.0 mmol) was added. The mixture was stirred at room
temperature for 3 h. Upon the completion of the reaction as
determined by TLC, the reaction mixture was poured onto ice-cold
water (150 mL). The mixture was stirred for another 10 minutes and
the solid was collected by filtration. The solid was washed with
water (50 mL) and dried by air. Then the solid was slurried in 5%
MeOH in DCM solution to afford desired product as a solid (200 mg,
30%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H),
8.21 (s, 1H), 7.31 (s, 1H), 7.09 (s, 1H), 5.86 (s, 1H), 4.26 (d,
J=4.4 Hz, 2H), 3.83 (d, J=9.60 Hz, 2H), 3.20-3.27 (m, 2H),
3.00-3.02 (m, 3H), 2.19 (s, 3H), 2.15 (s, 3H), 2.11 (s, 3H),
1.48-1.62 (m, 4H), 0.78 (t, J=6.8 Hz, 3H).
Step 8 to step 13: Synthesis of
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-[4-({1--
[4-(3-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]carbamoyl}-5-[e-
thyl(oxan-4-yl)amino]-4-methylphenyl)phenyl]-1,4,7,10,13,16-hexaoxaoctadec-
an-18-yl}carbamoyl)-2-methoxyphenyl]-5-(2,2-dimethylpropyl)pyrrolidine-2-c-
arboxamide (A2844)
[0431] Reactions in step 8 through step 10 were carried out using
the standard procedure of tosylation on hydroxyl group, tosyl group
displacement by bis-Boc-amine under potassium carbonate condition
and tosyl group displacement by phenol. The Suzuki coupling in Step
11 was carried out using palladium tetrakis(triphenylphosphine)
under the stand Suzuki coupling condition. The final two steps in
forming A2844 were followed the same procedure as described for the
synthesis of A1717. Compound A2844 was isolated as a solid.
[0432] .sup.1H NMR (400 MHz, CD.sub.3OD, ppm) .delta. 8.41-8.36 (m,
1H), 7.78-7.69 (m, 1H), 7.59-7.46 (m, 4H), 7.45-7.33 (m, 4H),
7.32-7.22 (m, 3H), 7.02-6.97 (m, 2H), 6.11 (s, 1H), 4.79-4.76 (m,
1H), 4.65-4.61 (m, 1H), 4.51 (s, 2H), 4.18-4.13 (m, 2H), 4.11-4.05
(m, 1H), 3.99 (s, 3H), 3.98-3.88 (m, 2H), 3.85-3.3.79 (m, 2H),
3.71-3.53 (m, 20H), 3.42-3.31 (m, 2H), 3.18-3.03 (m, 3H), 2.40 (s,
3H), 2.32 (s, 3H), 2.25 (s, 3H), 1.79-1.62 (m, 5H), 1.41-1.32 (m,
1H), 1.05 (s, 9H), 0.92-0.88 (t, J=6.8 Hz, 3H).
[0433] LC-MS calcd for
C.sub.72H.sub.87Cl.sub.2F.sub.2N.sub.7O.sub.12 (m/z) 1351.40, obsd
1352.70 (M+H.sup.+); t.sub.R=2.15 min (3.0 minute run).
[0434] Compound A2790 was prepared using the same method as
described for the preparation of A2844.
Example 9
Preparation of A2766
##STR00123## ##STR00124## ##STR00125##
[0435] Step 1: Synthesis of
[(2E)-3-(dimethylamino)-2-[2-(methylsulfanyl)pyrimidin-4-yl]prop-2-en-1-y-
lidene]dimethylazanium
[0436] Into a 100 mL 3-necked round-bottom flask, was placed a
solution of oxalyl dichloride (6.6 mL, 2.00 equiv) in
chloroform/N,N-dimethylformamide (45/6 mL) at 0.degree. C. The
above mixture was stirred for 30 min at 45.degree. C. The reaction
was cooled to 0.degree. C. 4-methyl-2-(methylsulfanyl)pyrimidine
(5.0 g, 142.65 mmol, 1.00 equiv) was added to the solution
separately at 0.degree. C. The resulting solution was stirred for
16 h at 45.degree. C. in an oil bath. The solids were collected by
filtration. This resulted in 8.5 g (90%) of
[(2E)-3-(dimethylamino)-2-[2-(methylsulfanyl)pyrimidin-4-yl]prop-
-2-en-1-ylidene]-dimethylazanium as a yellow solid.
[0437] LC-MS (ES.sup.+): m/z 250.95 [M+H.sup.+], t.sub.R=0.38 min,
(1.9 minute run).
Step 2: Synthesis of
2-(methylsulfanyl)-4-(1,2-oxazol-4-yl)pyrimidine
[0438] Into a 1000 mL 3-necked round-bottom flask, was placed a
solution of hydroxylamine hydrogen chloride (6.4 g, 92.75 mmol,
3.00 equiv) in water (300 mL), sodium carbonate (14.3 g, 134.92
mmol, 4.40 equiv),
[(2E)-3-(dimethylamino)-2-[2-(methylsulfanyl)pyrimidin-4-yl]prop-2-en-1-y-
lidene]dimethylazanium (7.7 g, 30.63 mmol, 1.00 equiv). The
resulting solution was stirred for 5 h at room temperature. The
solids were collected by filtration, concentrated under vacuum.
This resulted in 2.9 g (49%) of
2-(methylsulfanyl)-4-(1,2-oxazol-4-yl)pyrimidine as a brown
solid.
[0439] LC-MS (ES.sup.+): m/z 193.95[MH.sup.+], t.sub.R=1.22 min,
(2.6 minute run).
Step 3: Synthesis of
2-[2-(methylsulfanyl)pyrimidin-4-yl]-3-oxopropanenitrile
[0440] Into a 25-mL round-bottom flask, was placed a solution of
2-(methylsulfanyl)-4-(1,2-oxazol-4-yl)pyrimidine (1.0 g, 5.18 mmol,
1.00 equiv) in methanol/water (5/5 mL). Sodium hydroxide (210.0 mg,
5.25 mmol, 1.00 equiv) was added. The resulting solution was
stirred overnight at 70.degree. C. in an oil bath. The resulting
mixture was concentrated under vacuum. The pH value of the solution
was adjusted to 3-4 with hydrogen chloride. The solids were
collected by filtration, concentrated under vacuum. This resulted
in 1.0 g (100%) of
2-[2-(methylsulfanyl)pyrimidin-4-yl]-3-oxopropanenitrile as a brown
solid.
[0441] LC-MS (ES.sup.+): m/z 193.85[MH.sup.+], t.sub.R=0.48 min,
(1.9 minute run).
Step 4: Synthesis of
1-benzyl-4-[2-(methylsulfanyl)pyrimidin-4-yl]-1H-pyrazol-5-amine
[0442] Into a 100-mL round-bottom flask, was placed a solution of
2-[2-(methylsulfanyl)pyrimidin-4-yl]-3-oxopropanenitrile (1.0 g,
5.18 mmol, 1.00 equiv) in ethanol/3M hydrogen chloride (10/6 mL).
Benzylhydrazine hydrogen chloride (1.5 g, 7.73 mmol, 1.50 equiv)
was added. The resulting solution was stirred for 2 h at 83.degree.
C. in an oil bath. The resulting mixture was concentrated under
vacuum. The pH value of the solution was adjusted to 9 with sodium
carbonate. The resulting solution was extracted with
dichloromethane (20 mL.times.3) and the organic layers were
combined and concentrated under vacuum. The residue was applied
onto a silica gel column with ethyl acetate/petroleum ether (1:1).
This resulted in 900.0 mg (58%) of
1-benzyl-4-[2-(methylsulfanyl)pyrimidin-4-yl]-1H-pyrazol-5-amine as
a golden solid. LC-MS (ES.sup.+): m/z 297.90[MH.sup.+],
t.sub.R=0.83 min, (1.9 minute run).
Step 5: Synthesis of
1-benzyl-N,N-dimethyl-4-[2-(methylsulfanyl)pyrimidin-4-yl]-1H-pyrazol-5-a-
mine
[0443] Into a 25-mL round-bottom flask, was placed a solution of
1-benzyl-4-[2-(methylsulfanyl)pyrimidin-4-yl]-1H-pyrazol-5-amine
(150.0 mg, 0.50 mmol, 1.00 equiv) in tetrahydrofuran (10 mL),
sodium hydride (61.0 mg, 2.54 mmol, 3.00 equiv) was added to the
solution separately at 0.degree. C., 30 min later. To this mixture
iodomethane (0.4 mL, 10.00 equiv) was added. The resulting solution
was stirred for 12 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 were combined and concentrated under vacuum. The residue was
applied onto a silica gel column with ethyl acetate/petroleum ether
(1:2). This resulted in 76.0 mg (46%) of
1-benzyl-N,N-dimethyl-4-[2-(methylsulfanyl)pyrimidin-4-yl]-1H-pyrazol-5-a-
mine as light yellow oil. LC-MS (ES.sup.+): m/z 325.95 [M+H.sup.+],
t.sub.R=1.07 min, (1.9 minute run).
Step 6: Synthesis of
1-benzyl-4-(2-methanesulfonylpyrimidin-4-yl)-N,N-dimethyl-1H-pyrazol-5-am-
ine
[0444] Into a 25-mL round-bottom flask, was placed a solution of
1-benzyl-N,N-dimethyl-4-[2-(methylsulfanyl)pyrimidin-4-yl]-1H-pyrazol-5-a-
mine (76.0 mg, 0.23 mmol, 1.00 equiv) in dichloromethane (10 mL).
To this solution was added 3-chlorobenzoperoxoic acid (172.0 g,
996.70 mmol). 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
dichloromethane (20 mL.times.3) and the organic layers were
combined and concentrated under vacuum. This resulted in 76.0 mg
(91%) of
1-benzyl-4-(2-methanesulfonylpyrimidin-4-yl)-N,N-dimethyl-1H-pyrazol-5-am-
ine as a light yellow solid. LC-MS (ES.sup.+): m/z
358.00[MH.sup.+], t.sub.R=0.86 min, (1.9 minute run).
Step 7: Synthesis of
2-(2-(2-(1,4-trans)-4-aminocyclohexyloxy)-ethoxy)ethoxy)ethanol
[0445] To a solution of
2-(2-(2-((1,4-trans)-4-(dibenzylamino)cyclohexyloxy)-ethoxy)-ethoxy)-etha-
nol (512.0 mg, 1.2 mol, 1.0 equiv) in 20 mL MeOH was added Pd/C
(10%, 500 mg) under nitrogen atmosphere in a 100 mL round bottom
flask. The reaction flask was vacuumed and charged with a hydrogen
balloon. The reaction mixture was stirred for 16 h at 50.degree. C.
under hydrogen atmosphere. After the reaction was done, the
reaction mixture was filtered through a Celite pad and the filtrate
was concentrated under reduced pressure. This resulted in 247 mg of
2-[2-(2-[[(1,4-trans)-4-aminocyclohexyl]oxy]ethoxy)ethoxy]ethan-1-ol
as colorless oil. LC-MS (ES.sup.+): m/z 248.10 [M+H.sup.+],
t.sub.R=0.55 min, (2.6 minute run).
Step 8: Synthesis of
2-(2-(2-((1,4-trans)-4-(4-(1-benzyl-5-(dimethylamino)-1H-pyrazol-4-yl)pyr-
imidin-2-ylamino)cyclohexyloxy)ethoxy)ethoxy)ethanol
[0446] Into a 20 mL microwave vial, was placed a solution of
2-[2-(2-[[(1,4-trans)-4-aminocyclohexyl]oxy]ethoxy)ethoxy]ethan-1-ol
(247.0 mg, 1.0 mmol, 1.0 equiv) in iso-propanol (3.0 mL),
1-benzyl-4-(2-methanesulfonylpyrimidin-4-yl)-N,N-dimethyl-1H-pyrazol-5-am-
ine (357.0 mg, 1.0 mmol, 1.0 equiv), N,N-Diisopropylethylamine
(516.0 mg, 4.0 mmol, 4.0 equiv). The vial was irradiated in a
microwave at 130.degree. C. for 6 h. The resulting mixture was
concentrated under vacuum. The residue was applied onto a silica
gel column eluting with ethyl acetate/petroleum ether (1:1). This
resulted in 230.0 mg (44%) of
2-[2-(2-[[(1,4-trans)-4-([4-[1-benzyl-5-(dimethylamino)-1H-pyrazol-4-yl]p-
yrimidin-2-yl]amino)cyclohexyl]-oxy]ethoxy)ethoxy]ethan-1-ol as
colorless oil. LC-MS (ES.sup.+): m/z 525.10 [M+H.sup.+],
t.sub.R=0.75 min, (2.0 minute run).
Step 9: Synthesis of
2-[2-(2-[[(1,4-trans)-4-([4-[1-benzyl-5-(dimethylamino)-1H-pyrazol-4-yl]p-
yrimidin-2-yl]amino)cyclohexyl]oxy]ethoxy)ethoxy]ethyl
4-methylbenzene-1-sulfonate
[0447] Into a 100-mL round-bottom flask, was placed a solution of
2-[2-(2-[[(1,4-trans)-4-([4-[1-benzyl-5-(dimethylamino)-1H-pyrazol-4-yl]p-
yrimidin-2-yl]amino)cyclohexyl]oxy]ethoxy)ethoxy]ethan-1-ol (105.0
mg, 0.2 mmol, 1.0 equiv) in dichloromethane (20.0 mL),
triethylamine (40.0 mg, 0.4 mmol, 2.0 equiv),
4-dimethylaminopyridine (12.0 mg, 0.10 mmol, 0.1 equiv), 4-toluene
sulfonyl chloride (57.0 mg, 0.3 mmol, 1.5 equiv). The resulting
solution was stirred for 4 h at 40.degree. C. in an oil bath. The
resulting solution was quenched with 15 ml of water and extracted
with dichloromethane (20 mL.times.2). The combined organic layers
was dried over anhydrous sodium sulfate and concentrated under
vacuum. The residue was applied onto a silica gel column eluting
with ethyl acetate/petroleum ether (1:1). This resulted in 110 mg
(81%) of
2-[2-(2-[[(1,4-trans)-4-([4-[1-benzyl-5-(dimethylamino)-1H-pyrazol-4-yl]p-
yrimidin-2-yl]amino)cyclohexyl]oxy]-ethoxy)ethoxy]ethyl
4-methylbenzene-1-sulfonate as colorless oil. LC-MS (ES.sup.+): m/z
679.35 [M+H.sup.+], t.sub.R=1.34 min, (2.0 minute run).
Step 10 through Step 12: Preparation of
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-di-
methylpropyl)-N-[2-methoxy-5-({2-[2-(2-{[(1r,4r)-4-({4-[1-benzyl-5-(dimeth-
ylamino)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)cyclohexyl]oxy}-ethoxy)ethox-
y]ethyl}carbamoyl)phenyl]pyrrolidine-2-carboxamide (A2766)
[0448] The preparation of A2766 from the intermediate prepared in
step 9 was carried out using the same method descried for the
preparation of A1717, namely, the conversion of the tosyl group to
amine and followed by amide formation with MDM2 ligand.
[0449] .sup.1H NMR (400 MHz, CD.sub.3OD, ppm) .delta. 8.43-8.38 (m,
1H), 8.21-8.15 (m, 1H), 7.95 (s, 1H), 7.75-7.65 (m, 1H), 7.58 (s,
1H), 7.51-7.45 (m, 1H), 7.41-7.28 (m, 10H), 6.88-6.83 (m, 1H), 5.33
(s, 2H), 4.78-4.73 (m, 1H), 4.65-4.56 (m, 1H), 4.11-4.05 (m, 1H),
3.99 (s, 3H), 3.98-3.85 (m, 1H), 3.72-3.3.67 (m, 6H), 3.66-3.58 (m,
6H), 2.79 (s, 6H), 2.11-2.01 (m, 4H), 1.74-1.65 (m, 1H), 1.41-1.23
(m, 6H), 1.03 (s, 9H).
[0450] LC-MS calcd for
C.sub.59H.sub.68Cl.sub.2F.sub.2N.sub.10O.sub.6 (m/z) 1120.47, obsd
1121.50 (M+H.sup.+); t.sub.R=3.40 min (5.0 minute run).
[0451] Compound A2720, A2791 and A2792 were prepared with the same
method as described for the preparation of A2766.
[0452] Protein Degradation Biological Assays
[0453] The following biological assays were performed to evaluate
the protein degradation in various cell types using representative
compounds disclosed. In each assay, cells were treated with varying
amounts of compounds encompassed by the current disclosure as shown
in the Table. The degradation of the following proteins were
evaluated: bromodomain-containing protein 4 (BRD4), androgen
receptor (AR), c-Myc, c-Jun N-terminal kinases (JNK), and enhancer
of zeste homolog 2 (EZH2).
[0454] BRD4 Western Blot
[0455] VCaP cells were chased from ATCC and cultured in Dulbecco's
Modified Eagle's Medium (ATCC), supplemented with 10% FBS (ATCC)
and Penicillin/Streptomycin (Life Technologies). DMSO control and
compound treatments (0.03 .mu.M to 1 .mu.M) were performed in
12-well plates for 16 h. cells were harvested, and lysed in RIPA
buffer (50 mM Tris pH8, 150 mM NaCl, 1% Tx-100, 0.1% SDS, 0.5%
sodium deoxycholate) supplemented with protease and phosphatase
inhibitors. Lysates were clarified at 16,000 g for 10 minutes, and
protein concentration was determined. Equal amount of protein (20
.mu.g) was subjected to SDS-PAGE analysis and followed by
immunoblotting according to standard protocols. The antibodies used
were BRD4 (Cell signaling #13440), and Actin (Sigma #5441).
Detection reagents were Clarity Western ELC substrate (Bio-rad
#170-5060).
[0456] AR ELISA Assay
[0457] VCaP cells were chased from ATCC and cultured in Dulbecco's
Modified Eagle's Medium (ATCC), supplemented with 10% FBS (ATCC)
and Penicillin/Streptomycin (Life Technologies). DMSO control and
compound treatments (0.0001 .mu.M to 1 .mu.M) were performed in
96-well plates for 16 h. cells were harvested, and lysed with Cell
Lysis Buffer (catalog #9803) (20 mM Tris-HCl, pH 7.5, 150 mM NaCl,
1 mM Na.sub.2EDTA, 1 mM EGTA, 1% Triton, 2.5 mM sodium
pyrophosphate, 1 mM B-glycerophosphate, 1 mM Na.sub.3VO.sub.4, 1
.mu.g/mL leupeptin. Lysates were clarified at 16,000 g for 10
minutes, and loaded into PathScan AR ELISA (Cell Signaling Catalog
#12850). The PathScan Total Androgen Receptor Sanwich ELISA Kit is
a solid phase sandwich enzyme-linked immunosorbent assay (ELISA)
that detects endogenous levels of total androgen receptor protein.
An Androgen Receptor Rabbit mAb has been coated onto the
microwells. After incubation with cell lysates, androgen receptor
protein is captured by the coated antibody. Following extensive
washing, an Androgen receptor Mouse Detection mAbis added to detect
the captured androgen receptor protein. Anti-mouse IgG, HRP-linked
Antibody is then used to recognize the bound detection antibody.
HRP substrate, TMB, is added to develop color. The magnitude of
absorbance for the developed color is proportional to the quantity
of total androgen receptor protein.
[0458] c-Myc ELISA Assay
[0459] 22Rv-1 cells were purchased from ATCC and cultured in RPMI
with 10% FBS. Cells were harvested using trypsin (Gibco
#25200-114), counted and seeded at 30,000 cells/well at a volume of
75 .mu.L/well in RPMI with 10% FBS in 96-well plates. Cells were
dosed with compounds diluted in 0.1% DMSO, incubated for 18 h, then
washed and lysed in 50 .mu.L RIPA buffer (50 mM Tris pH 8, 150 mM
NaCl, 1% Tx-100, 0.1% SDS, 0.5% sodium deoxycholate) supplemented
with protease and phosphatase inhibitors. The lysates were
clarified at 4000 rpm at 4.degree. C. for 10 minutes. Aliquots were
added into a 96-well ELISA plate of Novex Human c-Myc ELISA kit
from Life Technologies (catalog # KH02041). Into each well was
added 50 .mu.L of c-Myc detection antibody. Plates were incubated
at room temperature for 3 h, washed with ELISA wash buffer,
followed by addition of 100 .mu.L of the anti-rabbit IgG-HRP
secondary antibody and 30 minutes of incubation. The plates were
washed with ELISA wash buffer followed by addition of 100 .mu.L of
TMB to each well. Color change was monitored every 5 minutes. Stop
solution (100 .mu.L) was added and plates were read at 450 nM.
[0460] JNK and EZH2 Western Blot Assay
[0461] Cells were purchased from ATCC and cultured in Dulbecco's
Modified Eagle's Medium (ATCC), supplemented with 10% FBS (ATCC)
and Penicillin/Streptomycin (Life Technologies). DMSO control and
compound treatments (0.003 .mu.M, 0.01 .mu.M, 0.03 .mu.M and 0.1
.mu.M) were performed in 12-well plates for 16 h. Cells were
harvested, and lysed in RIPA buffer (50 mM Tris pH8, 150 mM NaCl,
1% Tx-100, 0.1% SDS, 0.5% sodium deoxycholate) supplemented with
protease and phosphatase inhibitors. Lysates were clarified at
16,000 g for 10 minutes, and protein concentration was determined.
Equal amount of protein (20 .mu.g) was subjected to SDS-PAGE
analysis and followed by immunoblotting according to standard
protocols.
[0462] Synthesized molecules were assayed for protein degradation,
suppression, and regulation and growth inhibition of cancer cells
(FIG. 1). c-Myc suppression was observed in 22rv1 cells by chimeric
molecules, where BRD4 ligand is connected through linkers to MDM2
ligands using partial structural motif in RG7388. Chimeric
molecules with inactive MDM2 ligand (enantiomer of the active
counterpart) demonstrated no c-Myc suppression across a range of
concentrations, while chimeric molecules with active MDM2 ligand
showed dose dependent c-Myc suppression, suggesting BRD4
degradation mediated by MDM2 E3 ligase ubiquitination mechanism, as
c-Myc is directly regulated by the level of BRD4. Chimeric
molecules with MDM2 ligand as a racemate displayed similar c-Myc
suppression as observed in those containing active MDM2 ligand.
[0463] Western blot of HCT116 cells treated with chimeric molecules
was performed, where BRD4 ligand is connected through linkers to
MDM2 ligands using partial structural motif in RG7388 (FIG. 2).
Chimeric molecules with inactive MDM2 ligand (A-1891, A-1894)
demonstrated no p53 level increase and no MDM2 up-regulation, while
chimeric molecules with active MDM2 ligand (A-1864, A1892 and
A-1893, A-1877 carried a racemic MDM2 binding ligand) showed dose
dependent p53 level increase and up-regulation of MDM2, suggesting
chimeric molecules with BRD4 binding fragment and MDM2 binding
fragment connected through a linker can function as small molecule
MDM2 antagonist in stabilizing p53. The less significant MDM2 up
regulation and p53 level increase is due to the chimeric molecule
action mechanism of not only binding to MDM2 to block p53-MDM2
interaction but also degrading MDM2. Therefore, the net MDM2
up-regulation is significantly less, which also translated to p53
level due to MDM2-p53 feedback loop.
[0464] Western blot of HCT116 cells treated with chimeric molecules
(FIG. 3), where MDM2 ligand (using partial structural motif of
RG7388) is connected through linkers to VHL ligand. Chimeric
molecules with inactive MDM2 ligand (A-1897, A1908, and A-1911)
demonstrated no p53 level increase and no MDM2 up-regulation, while
chimeric molecules with active MDM2 ligand (A-1896, A-1907, and
A-1910, with A-1877, A-1895, and A-1909 carrying a racemic MDM2
binding ligand) showed dose dependent p53 level increase.
[0465] In p53.sup.WT HCT-116 colon cancer cell lines,
MDM2-recruiting BRD-4 PROTAC with active MDM2 binding moiety
(A-1893) caused very potent growth inhibition in comparison with
the MDM2-recruiting BRD-4 PROTAC with inactive MDM2 binding moiety
(A-1894) (FIG. 4). In this cell growth assay, BRD4-Cereblon PROTAC
A-825, MDM2 antagonist RG7388 (A-1850), the racemate of RG7388
(A-1851) and JQ1 were included as a direct comparison.
[0466] Time course of BRD4 degradation caused by BRD4-MDM2 chimeric
compound (A-1893) in human colon cancer cell line HCT116 (FIG. 5)
and human lung cancer cell line A549. (FIG. 6)
[0467] Following table is a representative of the degradation
activity of some exemplary compounds. The degradation activities
for target proteins are categorized as following: A (0 to 25%
degradation at 1 .mu.M); B (25 to 50% degradation at 1 .mu.M) and C
(larger than 50% degradation at 1 .mu.M).
TABLE-US-00001 Observed (m/z) Degradation activity Example from
LC/MS BRD4 AR JNK EZH2 Chemical Name A680 1082 A
4-(3-{4-[2-(2-{4-[2-(4-tert-butyl-2-
ethoxyphenyl)-4,5-bis(4-chlorophenyl)-
4,5-dimethyl-4,5-dihydro-1H-imidazole- 1-carbonyl]piperazin-1-
yl}ethoxy)ethoxy]phenyl}-4,4-dimethyl-
5-oxo-2-sulfanylideneimidazolidin-1-
yl)-2-(trifluoromethyl)benzonitrile A702 1260 B
4-(3-{4-[(17-{4-[2-(4-tert-butyl-2-
ethoxyphenyl)-4,5-bis(4-chlorophenyl)-
4,5-dimethyl-4,5-dihydro-1H-imidazole-
1-carbonyl]piperazin-1-yl}-3,6,9,12,15-
pentaoxaheptadecan-1-yl)oxy]phenyl}-4,4-
dimethyl-5-oxo-2-sulfanylideneimidazolidin-
1-yl)-2-(trifluoromethyl)benzonitrile A1283 1135, 1137 B
N-(17-{[3-(3-chloro-2-fluorophenyl)-4- (M + Na)
(4-chloro-2-fluorophenyl)-4-cyano-5-
(2,2-dimethylpropyl)pyrrolidin-2- yl]formamido}-3,6,9,12,15-
pentaoxaheptadecan-1-yl)-2-[(9S)-7-(4-
chlorophenyl)-4,5,13-trimethyl-3-thia-
1,8,11,12-tetraazatricyclo-[8.3.0.0.sup.2,.sup.6]trideca-
2(6),4,7,10,12-pentaen-9-yl]acetamide A1306 1025, 1027 B
N-(2-{2-[2-(2-{[3-(3-chloro-2-
fluorophenyl)-4-(4-chloro-2-fluorophenyl)-
4-cyano-5-(2,2-dimethylpropyl)pyrrolidin-2-
yl]formamido}ethoxy)ethoxy]ethoxy}ethyl)-
2-[(9S)-7-(4-chlorophenyl)-4,5,13- trimethyl-3-thia-1,8,11,12-
tetraazatricyclo[8.3.0.0.sup.2,.sup.6]trideca-
2(6),4,7,10,12-pentaen-9-yl]acetamide A1307 1069, 1071 C
N-(14-{[3-(3-chloro-2-fluorophenyl)-4-
(4-chloro-2-fluorophenyl)-4-cyano-5-
(2,2-dimethylpropyl)pyrrolidin-2- yl]formamido}-3,6,9,12-
tetraoxatetradecan-1-yl)-2-[(9S)-7-(4-
chlorophenyl)-4,5,13-trimethyl-3-thia- 1,8,11,12-tetraazatricyclo-
[8.3.0.0.sup.2,.sup.6]trideca-2(6),4,7,10,12-
pentaen-9-yl]acetamide A1571 934, 936 A
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-5-(2,2- dimethylpropyl)-N-{2-[2-(4-{[(1,3-
trans)-3-(3-chloro-4-cyanophenoxy)- 2,2,4,4-tetramethylcyclobutyl]
carbamoyl}phenoxy)ethoxy]ethyl}pyrrolidine- 2-carboxamide A1603
1024, 1026 A 3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-5-(2,2-
dimethylpropyl)-N-[1-(4-{[(1,3-trans)-3-
(3-chloro-4-cyanophenoxy)-2,2,4,4-
tetramethylcyclobutyl]carbamoyl}phenyl)-
1,4,7,10-tetraoxadodecan-12-yl]pyrrolidine- 2-carboxamide A1621
980, 982 A 3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-5-(2,2-
dimethylpropyl)-N-(2-{2-[2-(4-{[(1,3-
trans)-3-(3-chloro-4-cyanophenoxy)- 2,2,4,4-tetramethylcyclobutyl]
carbamoyl}phenoxy)ethoxy]ethoxy}ethyl)pyr- rolidine-2-carboxamide
A1688 1112, 1114 A 3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-5- (2,2-dimethylpropyl)-N-[1-(4-{[(1,3-
trans)-3-(3-chloro-4-cyanophenoxy)- 2,2,4,4-tetramethylcyclobutyl]-
carbamoyl}phenyl)-1,4,7,10,13,16-
hexaoxaoctadecan-18-yl]pyrrolidine-2- carboxamide A1717 1068, 1070
B 3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-5-(2,2- dimethylpropyl)-N-[1-(4-{[(1,3-
trans)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-
tetramethylcyclobutyl]carbamoyl}phenyl)-
1,4,7,10,13-pentaoxapentadecan-15- yl]pyrrolidine-2-carboxamide
A1720 1068, 1070 B (2R,3S,4R,5S)-3-(3-chloro-2-
fluorophenyl)-4-(4-chloro-2- fluorophenyl)-4-cyano-5-(2,2-
dimethylpropyl)-N-[1-(4-{[(1,3-trans)-3-
(3-chloro-4-cyanophenoxy)-2,2,4,4-
tetramethylcyclobutyl]carbamoyl}phenyl)-
1,4,7,10,13-pentaoxapentadecan-15- yl]pyrrolidine-2-carboxamide
A1735 1068, 1070 A (2S,3R,4S,5R)-3-(3-chloro-2-
fluorophenyl)-4-(4-chloro-2- fluorophenyl)-4-cyano-5-(2,2-
dimethylpropyl)-N-[1-(4-{[(1r,3r)-3-(3-
chloro-4-cyanophenoxy)-2,2,4,4-
tetramethylcyclobutyl]carbamoyl}phenyl)-
1,4,7,10,13-pentaoxapentadecan-15- yl]pyrrolidine-2-carboxamide
A1829 1174, 1176 B 3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-N-{4-[(2-{2-[2-(2-{2-
[(9S)-7-(4-chlorophenyl)-4,5,13- trimethyl-3-thia-1,8,11,12-
tetraazatricyclo[8.3.0.0.sup.2,.sup.6]trideca-
2(6),4,7,10,12-pentaen-9-
yl]acetamido}ethoxy)ethoxy]ethoxy}ethyl)carbamoyl]-
2-methoxyphenyl}-4-cyano-5-(2,2-
dimethylpropyl)pyrrolidine-2-carboxamide A1863 1130, 1132 B
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-N-[4-({2-[2-(2-{2- [(9S)-7-(4-chlorophenyl)-4,5,13-
trimethyl-3-thia-1,8,11,12-
tetraazatricyclo-[8.3.0.0.sup.2,.sup.6]trideca-
2(6),4,7,10,12-pentaen-9- yl]acetamido}ethoxy)ethoxy]-
ethyl}carbamoyl)-2-methoxyphenyl]-4-
cyano-5-(2,2-dimethylpropyl)pyrrolidine- 2-carboxamide A1864 1130,
1132 B (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-
4-(4-chloro-2-fluorophenyl)-N-[4-({2-
[2-(2-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-
trimethyl-3-thia-1,8,11,12-tetraazatricyclo-
[8.3.0.0.sup.2,.sup.6]trideca-2(6),4,7,10,12-pentaen-9-
yl]acetamido}ethoxy)ethoxy]ethyl}-
carbamoyl)-2-methoxyphenyl]-4-cyano-
5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide A1865 1130, 1132 A
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-N-[4-({2-[2-(2-{2-[(9S)-
7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-
tetraazatricyclo-[8.3.0.0.sup.2,.sup.6]trideca-2(6),4,7,10,12-
pentaen-9-yl]acetamido}ethoxy)ethoxy]ethyl}-
carbamoyl)-2-methoxyphenyl]-4-cyano-
5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide A1874 1172, 1174 B
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-N-{4-[(2-{2-[2-(2-{2-[(9S)-
7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-
tetraazatricyclo[8.3.0.0.sup.2,.sup.6]trideca-2(6),4,7,10,12-pentaen-
-9- yl]acetamido}ethoxy)ethoxy]ethoxy}ethyl)carbamoyl]-
2-methoxyphenyl}-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-
2-carboxamide A1875 1172, 1174 A
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-N-{4-[(2-{2-[2-(2-{2-
[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-
tetraazatricyclo[8.3.0.0.sup.2,.sup.6]trideca-2(6),4,7,10,12-pentaen-
-9- yl]acetamido}ethoxy)ethoxy]ethoxy}ethyl)carbamoyl]-
2-methoxyphenyl}-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-
2-carboxamide A1876 1216, 1218 C
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-
{4-[(14-{2-[(9S)-7-(4-chlorophenyl)-4,5,13-
trimethyl-3-thia-1,8,11,12-tetraazatricyclo[8.3.0.0.sup.2,.sup.6]-
trideca-2(6),4,7,10,12-pentaen-9-yl]acetamido}-3,6,9,12-
tetraoxatetradecan-1-yl)carbamoyl]-2-methoxyphenyl}-4-
cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide A1877 1173,
1175 3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N-[4-
({2-[2-({[(2S)-1-[(2S,4R)-4-hydroxy-2-
({[4-(4-methyl-1,3-thiazol-5-
yl)phenyl]methyl}carbamoyl)pyrrolidin-
1-yl]-3,3-dimethyl-1-oxobutan-2-
yl]carbamoyl}methoxy)ethoxy]ethyl}carbamoyl)-
2-methoxyphenyl]pyrrolidine-2-carboxamide A1890 1084, 1086 C
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-N-(4-{[2-(2-{2-[(9S)-7-
(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-
tetraazatricyclo[8.3.0.0.sup.2,.sup.6]-trideca-2(6),4,7,10,12-
pentaen-9-yl]acetamido}ethoxy)ethyl]carbamoyl}-
2-methoxyphenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-
2-carboxamide A1891 1084, 1086 A
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-N-(4-{[2-(2-{2-[(9S)-7-
(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-
tetraazatricyclo[8.3.0.0.sup.2,.sup.6]-trideca-2(6),4,7,10,12-
pentaen-9-yl]acetamido}ethoxy)ethyl]carbamoyl}-
2-methoxyphenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-
2-carboxamide A1892 1084, 1086 C
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-N-(4-{[2-(2-{2-[(9S)-7-
(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-
tetraazatricyclo[8.3.0.0.sup.2,.sup.6]-trideca-2(6),4,7,10,12-
pentaen-9-yl]acetamido}ethoxy)ethyl]carbamoyl}-
2-methoxyphenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-
2-carboxamide A1893 1216, 1218 C
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-N-{4-[(14-{2-[(9S)-7-(4-
chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-
tetraazatricyclo[8.3.0.0.sup.2,.sup.6]-trideca-2(6),4,7,10,12-
pentaen-9-yl]acetamido}-3,6,9,12-tetraoxatetradecan-1-
yl)carbamoyl]-2-methoxyphenyl}-4-cyano-5-(2,2-
dimethylpropyl)pyrrolidine-2-carboxamide A1894 1216, 1218 A
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-N-{4-[(14-{2-[(9S)-7-(4-
chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-
tetraazatricyclo[8.3.0.0.sup.2,.sup.6]-trideca-2(6),4,7,10,12-
pentaen-9-yl]acetamido}-3,6,9,12-tetraoxatetradecan-
1-yl)carbamoyl]-2-methoxyphenyl}-4-cyano-5-(2,2-
dimethylpropyl)pyrrolidine-2-carboxamide A1895 1261, 1263
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-
4-cyano-5-(2,2-dimethylpropyl)-N-{4-[(1-{[(2S)-
1-[(2S,4R)4-hydroxy-2-({[4-(4-methyl-1,3-
thiazol-5-yl)phenyl]methyl}carbamoyl)pyrrolidin-
1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl}-
2,5,8,11-tetraoxatridecan-13-yl)carbamoyl]-2-
methoxyphenyl}pyrrolidine-2-carboxamide A1896 1261, 1263
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N-{4-
[(1-{[(2S)-1-[(2S,4R)-4-hydroxy-2-
({[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl}-
carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-
yl]carbamoyl}-2,5,8,11-tetraoxatridecan-13-yl)carbamoyl]-2-
methoxyphenyl}pyrrolidine-2-carboxamide A1897 1261, 1263
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N-{4-
[(1-{[(2S)-1-[(2S,4R)-4-hydroxy-2-({[4-
(4-methyl-1,3-thiazol-5-yl)phenyl]methyl}-
carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-
yl]carbamoyl}-2,5,8,11-tetraoxatridecan-13-
yl)carbamoyl]-2-methoxyphenyl}pyrrolidine- 2-carboxamide A1907
1173, 1175 (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N-[4-({2-
[2-({[(2S)-1-[(2S,4R)-4-hydroxy-2-({[4-
(4-methyl-1,3-thiazol-5-yl)phenyl]methyl}-
carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-
yl]carbamoyl}methoxy)-ethoxy]ethyl}carbamoyl)-2-
methoxyphenyl]pyrrolidine-2-carboxamide A1908 1173, 1175
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N-[4-({2-
[2-({[(2S)-1-[(2S,4R)-4-hydroxy-2-({[4-
(4-methyl-1,3-thiazol-5-yl)phenyl]methyl}-
carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-
yl]carbamoyl}methoxy)-ethoxy]ethyl}carbamoyl)-2-
methoxyphenyl]pyrrolidine-2-carboxamide A1909 1217, 1219
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N-
{4-[(2-{2-[2-({[(2S)-1-[(2S,4R)-
4-hydroxy-2-({[4-(4-methyl-1,3-thiazol-5-
yl)phenyl]methyl}carbamoyl)pyrrolidin-
1-yl]-3,3-dimethyl-1-oxobutan-2-
yl]carbamoyl}methoxy)ethoxy]ethoxy}ethyl)carbamoyl]-
2-methoxyphenyl}-pyrrolidine-2-carboxamide A1910 1217, 1219
(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-
(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-
dimethylpropyl)-N-{4-[(2-{2-[2-({[(2S)-
1-[(2S,4R)-4-hydroxy-2-({[4-(4-methyl-1,3-thiazol-
5-yl)phenyl]methyl}-carbamoyl)pyrrolidin-1-yl]-
3,3-dimethyl-1-oxobutan-2-yl]carbamoyl}methoxy)-
ethoxy]ethoxy}ethyl)carbamoyl]-2-
methoxyphenyl}pyrrolidine-2-carboxamide A1911 1217, 1219
(2S,3R,4S,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N-{4-
[(2-{2-[2-({[(2S)-1-[(2S,4R)-
4-hydroxy-2-({[4-(4-methyl-1,3-thiazol-5-
yl)phenyl]methyl}-carbamoyl)pyrrolidin-1-yl]-
3,3-dimethyl-1-oxobutan-2-yl]carbamoyl}methoxy)-
ethoxy]ethoxy}ethyl)carbamoyl]-2-
methoxyphenyl}pyrrolidine-2-carboxamide A2434 997, 999 C
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-N-(3-{[5-(4-
{3-[4-cyano-3-(trifluoromethyl)phenyl]-
5,5-dimethyl-4-oxo-2-sulfanylideneimidazolidin-1-
yl}phenoxy)pentyl]oxy}propyl)-5-(2,2-
dimethylpropyl)pyrrolidine-2-carboxamide A2435 1146, 1148 C
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-N-{4-[(3-{[5-
(4-{3-[4-cyano-3-(trifluoromethyl)phenyl]-
5,5-dimethyl-4-oxo-2-sulfanylideneimidazolidin-1-
yl}phenoxy)pentyl]oxy}propyl)carbamoyl]-2-
methoxyphenyl}-5-(2,2-dimethylpropyl)pyrrolidine- 2-carboxamide
A2720 1077 A 3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N-(2-
methoxy-4-{[2-(2-{[(1,4-trans)-4-({4-
[1-benzyl-5-(dimethylamino)-1H-pyrazol-4-yl]pyrimidin-2-
yl}amino)cyclohexyl]oxy}ethoxy)ethyl]
carbamoyl}phenyl)pyrrolidine-2-carboxamide A2766 1121 A
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-
cyano-5-(2,2-dimethylpropyl)-N-[2-methoxy-5-({2-
[2-(2-{[(1,4-trans)-4-({4-[1-benzyl-5-
(dimethylamino)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)cyclo-
hexyl]oxy}ethoxy)ethoxy]ethyl}carbamoyl)phenyl]pyr-
rolidine-2-carboxamide A2790 1264 A
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-N-[4-({1-[4-(3-
{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-
yl)methyl]carbamoyl}-5-[ethyl(oxan-
4-yl)amino]-4-methylphenyl)phenyl]-1,4,7,10-
tetraoxadodecan-12-yl}carbamoyl)-2-methoxyphenyl]-
5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide A2791 1165 A
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N-[2-
methoxy-4-({1-[(1,4-trans)-4-({4-[1-
benzyl-5-(dimethylamino)-1H-pyrazol-4-yl]pyrimidin-
2-yl}amino)cyclohexyl]-1,4,7,10-tetraoxadodecan-12-
yl}carbamoyl)phenyl]pyrrolidine-2-carboxamide A2792 1209 A
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-
fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N-[2-
methoxy-5-({1-[(1,4-trans)-4-({4-[1-
benzyl-5-(dimethylamino)-1H-pyrazol-4-yl]pyrimidin-2-
yl}amino)cyclohexyl]-1,4,7,10,13-pentaoxapentadecan-15-
yl}carbamoyl)phenyl]pyrrolidine-2-carboxamide A2844 1352 A
3-(3-chloro-2-fluorophenyl)-4-(4-chloro-
2-fluorophenyl)-4-cyano-N-[4-({1-[4-(3-
{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-
3-yl)methyl]carbamoyl}-5-[ethyl(oxan-
4-yl)amino]-4-methylphenyl)phenyl]-1,4,7,10,13,16-
hexaoxaoctadecan-18-yl}carbamoyl)-2-methoxyphenyl]-
5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide
* * * * *