U.S. patent number RE43,003 [Application Number 12/882,769] was granted by the patent office on 2011-12-06 for epothilone derivatives.
This patent grant is currently assigned to Bristol-Myers Squibb Company. Invention is credited to James A. Johnson, Soong-Hoon Kim, Gregory D. Vite.
United States Patent |
RE43,003 |
Vite , et al. |
December 6, 2011 |
Epothilone derivatives
Abstract
The present invention relates to compounds of the formula
##STR00001## in which the variables G, W, Q, X, Y, B.sub.1,
B.sub.2, Z.sub.1, Z.sub.2, and R.sub.1 R.sub.7 are as defined
herein, methods for the preparation of the derivatives and
intermediates thereof.
Inventors: |
Vite; Gregory D. (Titusvile,
NJ), Kim; Soong-Hoon (Lawrenceville, NJ), Johnson; James
A. (Lawrenceville, NJ) |
Assignee: |
Bristol-Myers Squibb Company
(Princeton, NJ)
|
Family
ID: |
26729990 |
Appl.
No.: |
12/882,769 |
Filed: |
September 15, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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09084542 |
May 26, 1998 |
6605599 |
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60051951 |
Jul 8, 1997 |
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60067524 |
Dec 4, 1997 |
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Reissue of: |
11512623 |
Aug 30, 2006 |
7241755 |
Jul 10, 2007 |
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Current U.S.
Class: |
514/183; 549/355;
540/463; 540/462; 549/346; 548/202; 514/365; 514/366; 540/468;
548/203; 540/452; 548/204 |
Current CPC
Class: |
C07D
491/04 (20130101); C07D 493/04 (20130101); A61P
35/00 (20180101); C07D 491/044 (20130101); A61P
9/00 (20180101); A61P 19/00 (20180101); A61P
43/00 (20180101); C07D 417/06 (20130101) |
Current International
Class: |
C07D
417/06 (20060101); C07D 277/20 (20060101); C07D
277/26 (20060101) |
Field of
Search: |
;540/452,462,463,468
;548/202,203,204 ;549/346,355 ;514/183,365,366 |
References Cited
[Referenced By]
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|
Primary Examiner: Kifle; Bruck
Attorney, Agent or Firm: Winslow; Anastasia P.
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATIONS
The present application claims the priority benefit of U.S. patent
application Ser. No. 10/405,886, filed Apr. 3, 2003, which is a
continuation application of U.S. patent application Ser. No.
09/084,542, filed May 26, 1998, now U.S. Pat. No. 6,605,599, which
claims priority to provisional application No. 60/067,524, filed
Dec. 4, 1997, and provisional application No. 60/051,951, filed
Jul. 8, 1997.
Claims
What is claimed is:
1. A compound of the formula ##STR00049## wherein: ##STR00050## G
is selected from the group consisting of .[.alkyl; substituted
alkyl;.]. substituted aryl; a 4 to 7 membered monocyclic, 7 to 11
membered bicyclic, or 10 to 15 membered tricyclic saturated or
unsaturated ring system having between 1 and 3 heteroatoms selected
from nitrogen, oxygen, and sulfur; ##STR00051## .Iadd.and a
1-methyl-2-(substituted R') ethenyl group, wherein R' is a
monocyclic group selected from the group consisting of
pyrrolidinyl, pyrrolyl, indolyl, pyrazolyl, oxetanyl, pyrazolinyl,
imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl,
isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl,
isothiazolyl, isothiazolidinyl, furyl, tetrahydrofuryl, thienyl,
oxadiazolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl,
2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxazepinyl, azepinyl,
4-piperidonyl, pyridyl, N-oxo-pyridyl, pyrazinyl, pyrimidinyl,
pyridazinyl, tetrahydropyranyl, tetrahydrothiopyranyl,
tetraydrothiopyranyl sulfone, morpholinyl, thiomorpholinyl,
thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1,3-dioxolane
and tetrahyro-1,1-dioxothienyl, dioxanyl, isothiazolidinyl,
thietanyl, thiiranyl, triazinyl, and triazolyl; or a bicyclic
heterocyclic group selected from the group consisting of
benzothiazolyl, benzoxazoyl, benzothienyl, quinuclidinyl,
quinolinyl, quinolinyl-N-oxide, tetrahydroisoquinolinyl,
isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl,
benzofuryl, chromonyl, coumarinyl, cinnolinyl, quinoxalinyl,
indazolyl, pyrrolopyridyl, furopyridinyl, furo[2,3-c]pyridinyl,
furo[3,1-b]pyridinyl], furo[2,3-b]pyridinyl, dihydroisoindolyl,
dihydroquinazolinyl, 3,4-dihydro-4-oxo-quinazolinyl,
benzisothiazolyl, benzisoxazolyl, benzodiazinyl, benzofurazanyl,
benzothiopyranyl, benzotriazolyl, benzpyrazolyl, dihydrobenzofuryl,
dihydrobenzothienyl, dihydrobenzothiopyranyl,
dihydrobenzothiopyranyl sulfone, dihydrobenzopyranyl, indolinyl,
isochromanyl, isoindolinyl, naphthyridinyl, phthalazinyl,
piperonyl, purinyl, pyridopyridyl, quinazolinyl,
tetrahydroquinolinyl, thienofuryl, thienopyridyl, and
thienothienyl; wherein the R' substituents are selected from alkyl,
hydroxyalkyl, and oxo;.Iaddend. W is O or .[.NR.sub.15;.].
.Iadd.NH; .Iaddend. X is O or H, H; Y is selected from the group
consisting of O; H, OR.sub.16; OR.sub.17, OR.sub.17; NOR.sub.18; H,
NHOR.sub.19; H, NR.sub.20R.sub.21; H, H; and CHR.sub.22; wherein
OR.sub.17, OR.sub.17 can be a cyclic ketal; Z.sub.1 and Z.sub.2 are
independently CH.sub.2; .[.B.sub.1 and B.sub.2 are independently
selected from the group consisting of OR.sub.24, OCOR.sub.25, and
O--C(.dbd.O)--NR.sub.26R.sub.27, and when B.sub.1 is OH and Y is
OH, H they can form a six-membered ring ketal or acetal;.].
.Iadd.B.sub.1 is selected from the group consisting of OR.sub.24,
OCOR.sub.25, and O--C(.dbd.O)--NR.sub.26R.sub.27; B.sub.2 is
selected from the group consisting of OH, OCOR.sub.25, and
O--C(.dbd.O)--NR.sub.26R.sub.27; R.sub.1 and R.sub.2 are both
hydrogen;.Iaddend. .[.R.sub.1, R.sub.2,.]. R.sub.3, R.sub.4,
R.sub.5, R.sub.7, .[.R.sub.13, R.sub.14,.]. R.sub.18, R.sub.19,
R.sub.20, R.sub.21, R.sub.22, R.sub.26, and .Iadd.R.sub.27 are
.Iaddend.selected from the group consisting of H, alkyl,
substituted alkyl, and aryl.[., and when R.sub.1 and R.sub.2 are
alky, they can be joined to form a cycloalkyl;.]. .Iadd.;
.Iaddend.and when R.sub.3 and R.sub.4 are alkyl they can be joined
to form a cycloalkyl; R.sub.6 is methyl; R.sub.16, R.sub.17,
R.sub.24, and R.sub.25 are selected from the group consisting of H,
alkyl, and substituted alkyl; .[.R.sub.11, R.sub.12, R.sub.32, and
R.sub.33 are selected from the group consisting of H; alkyl;
substituted alkyl; aryl; substituted aryl; cycloalkyl containing 1
to 3 rings and 3 to 7 carbons per ring which may be further fused
with an unsaturated C.sub.3 C.sub.7 carbocyclic ring; and a 4 to 7
membered monocyclic, 7 to 11 membered bicyclic, or 10 to 15
membered tricyclic saturated or unsaturated ring system having
between 1 and 3 heteroatoms selected from nitrogen, oxygen, and
sulfur;.]. R.sub.8 is hydrogen or methyl; .[.R.sub.15 is selected
from the group consisting of H; alkyl; substituted alkyl; aryl;
substituted aryl; cycloalkyl containing 1 to 3 rings and 3 to 7
carbons per ring which may be further fused with an unsaturated
C.sub.3 C.sub.7 carbocyclic ring; a 4 to 7 membered monocyclic, 7
to 11 membered bicyclic, or 10 to 15 membered tricyclic saturated
or unsaturated ring system having between 1 and 3 heteroatoms
selected from nitrogen, oxygen, and sulfur; R.sub.32C.dbd.O,
R.sub.33SO2, hydroxy, O-alkyl or O-substituted alkyl;.]. or a
pharmaceutically acceptable salt thereof; with the proviso that
compounds wherein W and X are both O; and .[.R.sub.1, R.sub.2
and.]. R.sub.7 .[.are.]. .Iadd.is .Iaddend.H; and R.sub.3 and
R.sub.4 are methyl; and G is
1-methyl-2-(substituted)-4-thiazolyl-ethenyl; are excluded.
2. The compound of claim 1, wherein: X is O and Y is O; or a
pharmaceutically acceptable salt thereof.
3. The compound of claim 2, wherein: B.sub.1 is OH and B.sub.2 is
OH; or a pharmaceutically acceptable salt thereof.
4. The compound of claim 3, wherein W is O; or a pharmaceutically
acceptable salt thereof.
5. The compound of claim 4, wherein: .[.R.sub.1, R.sub.2, and.].
R.sub.7 .[.are.]. .Iadd.is .Iaddend.H; and R.sub.3, R.sub.4, and
R.sub.5 are methyl; or a pharmaceutically acceptable salt
thereof.
6. The compound of claim 5, wherein G is: ##STR00052## .Iadd.a
1-methyl-2-(substituted R') ethenyl group;.Iaddend. or a
pharmaceutically acceptable salt thereof.
7. The compound of claim 6, wherein .[.R.sub.11 is a 4 to 7
membered monocyclic saturated or unsaturated ring system having
from 1 and 3 heteroatoms selected from nitrogen, oxygen, and
sulfur;.]. .Iadd.R' is a monocyclic group selected from the group
consisting of pyrrolidinyl, pyrrol, indolyl, pyrazolyl, oxetanyl,
pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl,
oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl,
thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl,
tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl,
2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl,
2-oxazepinyl, azepinyl, 4-piperidonyl, pyridyl, N-oxo-pyridyl,
pyrazinyl, pyrimidinyl, pyridazinyl, tetrahydropyranyl,
tetrahydrothiopyranyl, tetrahydrothiopyranyl sulfone, morpholinyl,
thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl
sulfone, 1,3-dioxolane and tetrahydro-1,1-dioxothienyl, dioxanyl,
isothiazolidinyl, thietanyl, thiiranyl, triazinyl, and triazolyl;
.Iaddend.or a pharmaceutically acceptable salt thereof.
8. The compound of claim 3, wherein W is .[.NR.sub.15.].
.Iadd.NH.Iaddend.; or a pharmaceutically acceptable salt
thereof.
9. The compound of claim 8, wherein: .[.R.sub.1, R.sub.2, and.].
R.sub.7 .[.are.]. .Iadd.is .Iaddend.H; and R.sub.3, R.sub.4, and
R.sub.5 are methyl; or a pharmaceutically acceptable salt
thereof.
10. The compound of claim 9, wherein G is: ##STR00053## .Iadd.a
1-methyl-2-(substituted R') ethenyl group; .Iaddend. or a
pharmaceutically acceptable salt thereof.
11. The compound of claim 10, wherein .[.R.sub.11 is a 4 to 7
membered monocyclic saturated or unsaturated ring system having
from 1 and 3 heteroatoms selected from nitrogen, oxygen, and
sulfur;.]. .Iadd.R' is a monocyclic group selected from the group
consisting of pyrrolidinyl, pyrrol, indolyl, pyrazolyl, oxetanyl,
pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl,
oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl,
thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl,
tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl,
2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl,
2-oxazepinyl, azepinyl, 4-piperidonyl, pyridyl, N-oxo-pyridyl,
pyrazinyl, pyrimidinyl, pyridazinyl, tetrahydropyranyl,
tetrahydrothiopyranyl, tetrahydrothiopyranyl sulfone, morpholinyl,
thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl
sulfone, 1,3-dioxolane and tetrahydro-1,1-dioxothienyl, dioxanyl,
isothiazolidinyl, thietanyl, thiiranyl, triazinyl, and triazolyl;
.Iaddend.or a pharmaceutically acceptable salt thereof.
.[.12. The compound of claim 11, wherein R.sub.15 is H; or a
pharmaceutically acceptable salt thereof..].
13. A pharmaceutical composition comprising the compound of claim 1
or a pharmaceutically acceptable salt thereof; and a
pharmaceutically acceptable vehicle or diluent.
Description
FIELD OF THE INVENTION
The present invention relates to epothilone derivatives, methods
for the preparation of the derivatives and intermediates
therefor.
BACKGROUND OF THE INVENTION
Epothilones are macrolide compounds which find utility in the
pharmaceutical field. For example, Epothilones A and B having the
same structures:
##STR00002## have been found to exert microtubule-stabilizing
effects similar to TAXOL and hence cytotoxic activity against
rapidly proliferating cells, such as, tumor cells or other
hyperproliferative cellular disease, see Angew. Chem. Int. Ed.
Engl., 1996, 35, No. 13/14.
SUMMARY OF THE INVENTION
The present invention relates to compounds of the formula
##STR00003##
Q is selected from the group consisting of
##STR00004##
G is selected from the group consisting of alkyl, substituted
alkyl, substituted or unsubstituted aryl, heterocyclo,
##STR00005##
W is O or NR.sub.15;
X is O or H,H;
Y is selected from the group consisting of O; H,OR.sub.16;
OR.sub.17,OR.sub.17; NOR.sub.18; H,NOR.sub.19; H,NR.sub.20R.sub.21;
H,H; or CHR.sub.22; OR.sub.17OR.sub.17 can be a cyclic ketal;
Z.sub.1 and Z.sub.2 are selected from the group consisting of
CH.sub.2, O, NR.sub.23, S, or SO.sub.2, wherein only one of .[.Z.].
.Iadd.Z.sub.1 .Iaddend.and Z.sub.2 is a heteroatom;
B.sub.1 and B.sub.2 are selected from the group consisting of
OR.sub.24, or OCOR.sub.25, or O.sub.2CNR.sub.26R.sub.27; when
B.sub.1 is H and Y is OH, H they can form a six-membered ring ketal
or acetal;
D is selected from the group consisting of NR.sub.28R.sub.29,
NR.sub.30COR.sub.31 or saturated heterocycle;
R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7,
R.sub.13, R.sub.14, R.sub.18, R.sub.19, R.sub.20, R.sub.21,
R.sub.22, R.sub.26, and R.sub.27 are selected from the group H,
alkyl, substituted alkyl, or aryl and when R.sub.1 and R.sub.2 are
alkyl can be joined to form a cycloalkyl; R.sub.3 and R.sub.4 are
alkyl can be joined to form a cycloalkyl;
R.sub.9, R.sub.10, R.sub.16, R.sub.17, R.sub.24, R.sub.25, and
R.sub.31 are selected from the group H, alkyl, or substituted
alkyl;
R.sub.8, R.sub.11, R.sub.12, R.sub.28, R.sub.30, R.sub.32,
R.sub.33, and R.sub.30 are selected from the group consisting of H,
alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, or
heterocyclo;
R.sub.15, R.sub.23 and R.sub.29 are selected from the group
consisting of H, alkyl, substituted alkyl, aryl, substituted aryl,
cycloalkyl, heterocyclo, R.sub.32C.dbd.O, R.sub.33SO.sub.2,
hydroxy, O-alkyl or O-substituted alkyl;
and any salts, solvates or hydrates thereof.
Proviso
The present invention does not include compounds of formula V
wherein W and X are both O; and R.sub.1, R.sub.2, R.sub.7, are H;
and R.sub.3, R.sub.4, R.sub.6, are methyl; and R.sub.8, is H or
methyl; and Z.sub.1, and Z.sub.2, are CH.sub.2; and G is
1-methyl-2-(substituted-4-thiazolyl)ethenyl; and Q is as defined
above.
DETAILED DESCRIPTION OF THE INVENTION
Listed below are definitions of various terms used to describe this
invention. These definitions apply to the terms as they are used
throughout this specification, unless otherwise limited in specific
instances, either individually or as part of a larger group.
The term "alkyl" refers to straight or branched chain unsubstituted
hydrocarbon groups of 1 to 20 carbon atoms, preferably 1 to 7
carbon atoms. The expression "lower alkyl" refers to unsubstituted
alkyl groups of 1 to 4 carbon atoms.
The term "substituted alkyl" refers to an alkyl group substituted
by, for example, one to four substituents, such as, halo,
trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, cycloalkoxy,
heterocyclooxy, oxo, alkanoyl, aryloxy, alkanoyloxy, amino,
alkylamino, arylamino, aralkylamino, cycloalkylamino,
heterocycloamino, disubstituted amines in which the 2 amino
substituents are selected from alkyl, aryl or aralkyl,
alkanoylamino, aroylamino, aralkanoylamino, substituted
alkanoylamino, substituted arylamino, substituted aralkanoylamino,
thiol, alkylthio, arylthio, aralkylthio, cycloalkylthio,
heterocyclothio, alkylthiono, arylthiono, aralkylthiono,
alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, sulfonamido (e.g.
SO.sub.2NH.sub.2), substituted sulfonamido, nitro, cyano, carboxy,
carbamyl (e.g. CONH.sub.2), substituted carbamyl (e.g. CONH alkyl,
CONH aryl, CONH aralkyl or cases where there are two substituents
on the nitrogen selected from alkyl, aryl or aralkyl),
alkoxycarbonyl, aryl, substituted aryl, guanidino and heterocyclos,
such as, indolyl, imidazolyl, furyl, thienyl, thiazolyl,
pyrrolidyl, pyridyl, pyrimidyl and the like. Where noted above
where the substituent is further substituted it will be with
halogen, alkyl, alkoxy, aryl or aralkyl.
The term "halogen" or "halo" refers to fluorine, chlorine, bromine
and iodine.
The term "aryl" refers to monocyclic or bicyclic aromatic
hydrocarbon groups having 6 to 12 carbon atoms in the ring portion,
such as phenyl, naphthyl, biphenyl and diphenyl groups, each of
which may be substituted.
The term "aralkyl" refers to an aryl group bonded directly through
an alkyl group, such as benzyl.
The term "substituted aryl" refers to an aryl group substituted by,
for example, one to four substituents such as alkyl, substituted
alkyl, halo, trifluoromethoxy, trifluoromethyl, hydroxy, alkoxy,
cycloalkyloxy, heterocyclooxy, alkanoyl, alkanoyloxy, amino,
alkylamino, aralkylamino, cycloalkylamino, heterocycloamino,
dialkylamino, alkanoylamino, thiol, alkylthio, cycloalkylthio,
heterocyclothio, ureido, nitro, cyano, carboxy, carboxyalkyl,
carbamyl, alkoxycarbonyl, alkylthiono, arylthiono, alkysulfonyl,
sulfonamido, aryloxy and the like. The substituent may be further
substituted by halo, hydroxy, alkyl, alkoxy, aryl, substituted
aryl, substituted alkyl or aralkyl.
The term "cycloalkyl" refers to optionally substituted,
saturated-cyclic hydrocarbon ring systems, preferably containing 1
to 3 rings and 3 to 7 carbons per ring which may be further fused
with an unsaturated C.sub.3 C.sub.7 carbocyclic ring. Exemplary
groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, cyclooctyl, cyclodecyl, cyclododecyl, and adamantyl.
Exemplary substituents include one or more alkyl groups as
described above, or one or more groups described above as alkyl
substituents.
The terms "heterocycle", "heterocyclic" and "heterocyclo" refer to
an optionally substituted, fully saturated or unsaturated, aromatic
or nonaromatic cyclic group, for example, which is a 4 to 7
membered monocyclic, 7 to 11 membered bicyclic, or 10 to 15
membered tricyclic ring system, which has at least one heteroatom
in at least one carbon atom-containing ring. Each ring of the
heterocyclic group containing a heteroatom may have 1, 2 or 3
heteroatoms selected from nitrogen atoms, oxygen atoms and sulfur
atoms, where the nitrogen and sulfur heteroatoms may also
optionally be oxidized and the nitrogen heteroatoms may also
optionally be quaternized. The heterocyclic group may be attached
at any heteroatom or carbon atom.
Exemplary monocyclic heterocyclic groups include pyrrolidinyl,
pyrrolyl, indolyl, pyrazolyl, oxetanyl, pyrazolinyl, imidazolyl,
imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolinyl,
isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl, isothiazolyl,
isothiazolidinyl, furyl, tetrahydrofuryl, thienyl, oxadiazolyl,
piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl,
2-oxopyrrolidinyl, 2-oxazepinyl, azepinyl, 4-piperidonyl, pyridyl,
N-oxo-pyridyl, pyrazinyl, pymidinyl, pyridazinyl,
tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydrothiopyranyl
sulfone, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide,
thiomorpholinyl sulfone, 1,3-dioxolane and
tetrahydro-1,1-dioxothienyl, dioxanyl, isothiazolidinyl, thietanyl,
thiiranyl, triazinyl, and triazolyl, and the like.
Exemplary bicyclic heterocyclic groups include benzothiazolyl,
benzoxazolyl, benzothienyl, quinuclidinyl, quinolinyl,
quinolinyl-N-oxide, tetrahydroisoquinolinyl, isoquinolinyl,
benzimidazolyl, benzopyranyl, indolizinyl, benzofuryl, chromonyl,
coumarinyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl,
furopyridinyl (such as furo[2,3-c]pyridinyl, furo[3,1-b]pyridinyl]
or furo[2,3-b]pyridinyl), dihydroisoindolyl, dihydroquinazolinyl
(such as 3,4-dihydro-4-oxo-quinazolinyl), benzisothiazolyl,
benzisoxazolyl, benzodiazinyl, benzofurazanyl, benzothiopyranyl,
benzotriazolyl, benzpyrazolyl, dihydrobenzofuryl,
dihydrobenzothienyl, dihydrobenzothiopyranyl,
dihydrobenzothiopyranyl sulfone, dihydrobenzopyranyl, indolinyl,
isochromanyl, isoindolinyl, naphthyridinyl, phthalazinyl,
piperonyl, purinyl, pyridopyridyl, quinazolinyl,
tetrahydroquinolinyl, thienofuryl, thienopyridyl, thienothienyl,
and the like.
Exemplary substituents include one or more alkyl groups as
described above or one or more groups described above as alkyl
substituents. Also included are smaller heterocyclos, such as,
epoxides and aziridines.
The term "heteroatoms" shall include oxygen, sulfur and
nitrogen.
The compounds of formula V may form salts with alkali metals such
as sodium, potassium and lithium, with alkaline earth metals such
as calcium and magnesium, with organic bases such as
dicyclohexylamine, tributylamine, pyridine and amino acids such as
arginine, lysine and the like. Such salts can be obtained, for
example, by exchanging the carboxylic acid protons, if they contain
a carboxylic acid, in compounds of formula V with the desired ion
in a medium in which the salt precipitates or in an aqueous medium
followed by evaporation. Other salts can be formed as known to
those skilled in the art.
The compounds for formula V form salts with a variety of organic
and inorganic acids. Such salts include those formed with hydrogen
chloride, hydrogen bromide, methanesulfonic acid,
hydroxyethanesulfonic acid, sulfuric acid, acetic acid,
trifluoroacetic acid, maleic acid, benzenesulfonic acid,
toluenesulfonic acid and various others (e.g., nitrates,
phosphates, borates, tartrates, citrates, succinates, benzoates,
ascorbates, salicylates and the like). Such salts are formed by
reacting a compound of formula V in an equivalent amount of the
acid in a medium in which the salt precipitates or in an aqueous
medium followed by evaporation.
In addition, zwitterions ("inner salts") are formed.
Compounds of the formula V may also have prodrug forms. Any
compound that will be converted in vivo to provide the bioactive
agent (i.e., the compound for formula V) is a prodrug within the
scope and spirit of the invention.
For example compounds of the formula V may form a carboxylate ester
moiety. The carboxylate esters are conveniently formed by
esterifying any of the carboxylic acid functionalities found on the
disclosed ring structure(s).
Various forms of prodrugs are well known in the art. For examples
of such prodrug derivatives, see: a) Design of Prodrugs, edited by
H. Bundgaard, (Elsevier, 1985) and Methods in Enzymology, Vol. 112,
pp. 309 396, edited by K. Widder et al. (Academic Press, 1985); b)
A Textbook of Drug Design and Development, edited by
Krosgaard-Larsen and H. Bundgaard, Chapter 5, "Design and
Application of Prodrugs," by H. Bundgaard, pp. 113 191 (1991); c)
H. Bundgaard, Advanced Drug Delivery Reviews, 8, pp. 1 38 (1992);
d) H. Bundgaard et al., Journal of Pharmaceutical Sciences, 77, p.
285 (1988); and e) N. Kakeya et al., Chem. Phar. Bull., 32, p. 692
(1984).
It should further be understood that solvates (e.g., hydrates) of
the compounds of formula V are also within the scope of the present
invention. Methods of solvation are generally known in the art.
Use and Utility
The compounds of formula V are microtubule-stabilizing agents. They
are thus useful in the treatment of a variety of cancers or other
abnormal proliferative diseases, including (but not limited to) the
following; carcinoma, including that of the bladder, breast, colon,
kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid and
skin; including squamous cell carcinoma; hematopoietic tumors of
lymphoid lineage, including leukemia, acute lymphocytic leukemia,
acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma,
Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma and
Burketts lymphoma; hematopoietic tumors of myeloid lineage,
including acute and chronic myelogenous leukemias and promyelocytic
leukemia; tumors of mesenchymal origin, including fibrosarcoma and
rhabdomyoscarcoma; other tumors, including melanoma, seminoma,
teratocarcinoma, neuroblastoma and glioma; tumors of the central
and peripheral nervous system, including astrocytoma,
neuroblastoma, glioma, and schwannomas; tumors of mesenchymal
origin, including fibrosarcoma, rhabdomyoscaroma, and osteosarcoma;
and other tumors, including melanoma, xenoderma pigmentosum,
keratoacanthoma, seminoma, thyroid follicular cancer and
teratocarcinoma.
Compounds of formula V may also inhibit tumor angiogenesis, thereby
affecting abnormal cellular proliferation. Such anti-angiogenesis
properties of the compounds of formula V may also be useful in the
treatment of certain forms of blindness related to retinal
vascularization, arthritis, especially inflammatory arthritis,
multiple sclerosis, restinosis and psoriasis.
Compounds of formula V may induce or inhibit apoptosis, a
physiological cell death process critical for normal development
and homeostasis. Alterations of apoptotic pathways contribute to
the pathogenesis of a variety of human diseases. Compounds of
formula V, as modulators of apoptosis, will be useful in the
treatment of a variety of human diseases with aberrations in
apoptosis including cancer (particularly, but not limited to
follicular lymphomas, carcinomas with p53 mutations, hormone
dependent tumors of the breast, prostrate and ovary, and
precancerous lesions such as familial adenomatous polyposis), viral
infections (including but not limited to herpesvirus, poxvirus,
Epstein-Barr virus, Sindbis virus and adenovirus), autoimmune
diseases (including but not limited to systemic lupus
erythematosus, immune mediated glomerulonephritis, rheumatoid
arthritis, psoriasis, inflammatory bowel diseases and autoimmune
diabetes mellitus), neurodegenerative disorders (including but not
limited to Alzheimer's disease, AIDS-related dementia, Parkinson's
disease, amyotrophic lateral sclerosis, retinitis pigmentosa,
spinal muscular atrophy and cerebellar degeneration), AIDS,
myelodysplastic syndromes, aplastic anemia, ischemic injury
associated myocardial infarctions, stroke and reperfusion injury,
arrhythmia, atherosclerosis, toxin-induced or alcohol induced liver
diseases, hematological diseases (including but not limited to
chronic anemia and aplastic anemia), degenerative diseases of the
musculoskeletal system (including but not limited to osteoporosis
and arthritis), aspirin-sensitive rhinosinusitis, cystic fibrosis,
multiple sclerosis, kidney diseases, and cancer pain.
The compounds of this invention are also useful in combination with
known anti-cancer and cytotoxic agents and treatments, including
radiation. If formulated as a fixed dose, such combination products
employ the compounds of this invention within the dosage range
described below and the other pharmaceutically active agent within
its approved dosage range. Compounds of formula V can be used
sequentially with known anticancer or cytotoxic agents and
treatment, including radiation when a combination formulation is
inappropriate. Especially useful are cytotoxic drug combinations
wherein the second drug chosen acts in a different phase of the
cell cycle, e.g. S phase, than the present compounds of formula V
which exert their effects at the G.sub.2-M phase.
e.g.
Thymidilate Synthase Inhibitors DNA Cross Linking Agents
Topoisomerase I and II Inhibitors DNA Alkylating Agents
Ribonucleoside Reductase Inhibitors Cytotoxic Factors e.g.
TNF-alpha or Growth factor inhibitors e.g. HER 2 receptor MAB's
The present compounds may exist as multiple optical, geometric, and
stereoisomers. Included within the present invention are all such
isomers and mixtures thereof.
The compounds of this invention can be formulated with a
pharmaceutical vehicle or diluent for oral, intravenous or
subcutaneous administration. The pharmaceutical composition can be
formulated in a classical manner using solid or liquid vehicles,
diluents and additives appropriate to the desired mode of
administration. Orally, the compounds can be administered in the
form of tablets, capsules, granules, powders and the like. The
compounds are administered in a dosage range of about 0.05 to 200
mg/kg/day, preferably less than 100 mg/kg/day, in a single dose or
in 2 to 4 divided doses.
Preferred Compounds
Especially preferred compounds of formula V are those wherein Q
is:
##STR00006## X is O; Y is O; Z.sub.1 and Z.sub.2 are CH.sub.2; and
W is NR.sub.15.
Method of Preparation
Compounds of formula V are prepared by the following schemes.
##STR00007## ##STR00008## wherein R.sub.3, R.sub.4, R.sub.5,
R.sub.6, R.sub.8 and R.sub.15 are as above and P.sub.1 is an oxygen
protecting group.
Compounds of formula V where W is NR.sub.15 and X is O can be
prepared as outlined in Scheme 1. A compound of formula XII, where
P.sub.1 is an oxygen protecting group such as t-butyldimethylsilyl,
can be prepared from a compound of formula VI by known methods
(i.e., Nicolaou, K. C. et al., Angew. Chem. Int. Ed. Engl., 36:166
168 (1997)). Aldol reaction of a compound of formula XII and a
compound of formula XIV provides a compound of formula XIII. The
compound of formula XIV can be prepared by known methods (i.e.,
Schinzer, D. et al., Eur. Chem. Chron., 1:7 10 (1996)). An aldehyde
of formula XVIII can be prepared from a compound of formula XV as
shown in Scheme 1 or by using known methods (i.e., Taylor, R. E. et
al., Tetrahedron Lett., 38:2061 2064 (1997)). A compound of formula
XIX can be prepared from a compound XVIII by treatment with an
amine using dehydrating conditions such as catalytic
p-toluenesulfonic acid and azeotropic removal of water. A compound
of formula XX can be prepared from a compound of formula XIX by
treatment with an allylating reagent such as allylmagnesium
bromide. A compound of formula XXI can be prepared from compounds
of formulas XIII and XX, by standard amide bond coupling agents
(i.e., DCC, BOP, EDC/HOBT, PyBrOP). A compound of formula XXII can
be prepared from a compound of formula XXI by ring-closing
metathesis using either the Grubbs
(RuCl.sub.2(.dbd.CHPh)(PCY.sub.3).sub.2; see Grubbs, R. H. et al.,
Angew. Chem. Int. Ed. Engl., 34:2039 (1995)) or Schrock catalysts
(See Schrock, R. R. et al., J. Am. Chem. Soc., 112:3875 (1990)).
Deprotection of a compound of formula XXI using, for example when
P.sub.1 is a t-butyldimethylsily group, hydrogen fluoride in
acetronitrile or tetra-n-butyl ammonium fluoride in THF provides a
compound of formula V where Q is an ethylene group, W is NR.sub.15,
X is O, an R.sub.3, R.sub.4, R.sub.5, R.sub.6 are defined as
described above. Regioselective epoxidation of a compound of
formula V where Q is an ethylene group using dimethyldioxirane
provides a compound of formula V where Q is an oxirane group, W is
NR.sub.15, X is O, and R.sub.3, R.sub.4, R.sub.5, R.sub.15 are
defined as described above.
##STR00009##
Alternatively, a compound of formula VIII can be prepared by
reaction of a compound of formula XXIII with magnesium and an acid
chloride (R.sub.5CH.sub.2COCl) to give a compound of formula XXIV
(See for example: Heathcock, C. et al., J. Org. Chem., 55:1114 1117
(1990)), followed by ozononolysis to give a compound of formula
VIII as shown in Scheme 2.
##STR00010##
Alternatively, a compound of formula XIV can be prepared as shown
in Scheme 3. Reaction of a compound of formula XXV and
pseudoephedrine provides a compound of formula XXVI. A compound of
formula XXVII can be prepared from a compound of formula XXVI by
alkylation with a pentenyl halide such as 5-bromopentene according
to the method of Meyers (i.e., Meyers, A. et al., J. Am. Chem.
Soc., 116:9361 9362 (1994)). A compound of formula XXVIII can be
prepared from a compound of formula XXVII with a reducing agent
such as lithium pyrrolidinyl borohydride. Oxidation of a compound
of formula XXVIII, using for example pyridinium chlorochromate,
provides a compound of formula XIV. Direct conversion of a compound
of formula XXVII to a compound of formula XIV can be accomplished
with a reducing agent such as lithium triethoxy-aluminum
hydride.
##STR00011##
Alternatively, a compound of formula XX can be prepared from
allylglycine as shown in Scheme 4. Allylglycine can be N-protected
using methods known in the art to give a compound of formula XXIX,
where P.sub.2 is a suitable N-protecting group such as
t-butyloxycarbonyl. Optionally, where R.sub.29 is not hydrogen, a
compound of formula XXX can be prepared from a compound of formula
XXIX by alkylation with an alkyl halide in the presence of a base
such as sodium hydride. A compound of formula XXXI can be prepared
from a compound of formula XXX using N,O-dimethylhydroxylamine and
standard coupling agents such as EDCI and HOBT. A compound of
formula XXXII can be prepared from a hydroxamate XXXI by treatment
with an organometallic reagent such as an alkyl or arylmagnesium
halide. Wittig olefination of a compound of formula XXXII provides
a compound of formula XXXIII (the Wittig reagent is prepared as
reported: Danishefsky, S. E. et al., J. Org. Chem., 61:7998 7999
(1996)). N-Deprotection of a compound of formula XXXIII using
methods known in the art provides a compound of formula XX.
##STR00012##
A compound of formula V where W is NR.sub.15, X is oxygen, and G is
a 1,2-disubstituted olefin can be prepared as shown in Scheme 5. A
compound of formula XXXV can be prepared by Wittig olefination of a
compound of formula XXXII. A compound of formula XXXIV can be
prepared by methods known in the art. A compound of formula XXXVI
can be prepared by N-deprotection of a compound of formula XXXV
using methods known in the art. A compound of formula XXXVII can be
prepared by coupling reaction of a compound of formula XXXVI and a
compound of formula XIII using standard coupling agents such as
EDCI and HOBT. A compound of formula XXXVIII can be prepared from a
compound of formula XXXVII by methods described in Scheme 1 for the
preparation of a compound of formula XXII. Using methods described
in Scheme 1 (steps o and p), a compound of formula XXXVIII can be
converted to compounds of formula V where W is NR.sub.15, X is
oxygen, and G is a 1,2-disubstituted olefin.
##STR00013##
A compound of formula V where both W and X are oxygen, and G is a
1,2-disubstituted olefin can be prepared as shown in Scheme 6. A
compound of formula XXXX can be prepared from a compound of formula
XXXIX by treatment with an allylating agent such as allylmagnesium
bromide. Enantiomerically pure XXXX can be prepared by employing
chiral reagents (see, for example: Taylor, R. E. et al.,
Tetrahedron Lett., 38:2061 2064 (1997); Nicolaou, K. C. et al.,
Angew. Chem. Int. Ed. Engl., 36:166 168 (1997); Keck, G. et al., J.
Am. Chem. Soc., 115:8467 (1993)). A compound of formula XXXXI can
be prepared from compounds of formula XXXX and XIII by using
standard esterification methods such as DCC and DMAP. A compound of
formula XXXXII can be prepared from a compound of formula XXXXI via
ring-closing olefin metathesis as described in Scheme 1 for the
preparation of a compound of formula XXII. Compounds of formula V
where both W and X are oxygen, and G is a 1,2-disubstituted olefin
can be prepared from a compound of formula XXXXII by deprotection
(where Q is an ethylene group) and, if desired, epoxidation (where
Q is an oxirane group) as described above.
##STR00014##
A compound of formula V where both W and X are oxygen, and G is
alkyl, substituted alkyl, aryl, heteroaryl, bicycloaryl, or
bicycloheteroaryl can be prepared as shown in Scheme 7. A compound
of formula XXXXIV can be prepared by allylation of a compound of
formula XXXXIII, where G is alkyl, substituted alkyl, aryl,
heteroaryl, bicycloaryl, or bicycloheteroaryl, by reaction with an
allylating reagent such as allyl magnesium bromide. A compound of
formula XXXXV can be prepared from a compound of formula XXXXIV via
esterification with a compound of formula XIII using, for example,
DCC and DMAP. A compound of formula XXXXVI can be prepared from a
compound of formula XXXXV by ring-closing metathesis as described
above. Following the methods outlined above for Scheme 1, a
compound of formula XXXXVI can be converted to compounds of formula
V by deprotection and subsequent epoxidation.
##STR00015##
A compound of formula V where W is NR.sub.15, X is oxygen, and G is
alkyl, substituted alkyl, aryl, heteroaryl, bicycloaryl, or
bicycloheteroaryl can be prepared as shown in Scheme 8. A compound
of formula XXXXVII can be prepared by reaction of a compound of
formula XXIII, where G is alkyl, substituted alkyl, aryl,
heteroaryl, bicycloaryl, or bicycloheteroaryl, and an amine under
dehydrating conditions. A compound of formula XXXXVIII can be
prepared from a compound of formula XXXXVII by treatment with an
allylating agent such as allylmagnesium bromide. A compound of
formula XXXXIX can be prepared from a compound of formula XXXXVIII
and a compound of formula XIII by standard amide bond coupling
techniques using, for example, EDCI and HOBT. A compound of formula
L can be prepared from a compound of formula XXXXIX by ring-closing
metathesis as described above. Following the methods outlined above
for Scheme 1, a compound of formula L can be converted to compounds
of formulas V by deprotection and subsequent epoxidation.
##STR00016##
A compound of formula V where X is oxygen, W is NR.sub.15, and G
is
##STR00017## and D is selected from the group consisting of
NR.sub.28R.sub.29, NR.sub.30COR.sub.31, and saturated heterocycle
(i.e., piperidinyl, morpholinyl, piperazinyl, etc.) can be prepared
as shown in Scheme 9. A compound of formula LI can be prepared from
a compound of formula XXXII by reductive amination using a primary
or secondary amine and a reducing agent such as sodium
triacetoxyborohydride. Compounds of formula LIII, LIV, and V can
then be prepared following methods described above in Scheme 1.
##STR00018##
Alternatively, a compound of formula V where X is oxygen, W is
oxygen or NR.sub.15, and G is
##STR00019## and D is selected from the group consisting of
NR.sub.28R.sub.29, NR.sub.30COR.sub.31, and saturated heterocycle
(i.e., piperidinyl, morpholinyl, piperazinyl, etc.) can be prepared
from a compound of formula V as shown in Scheme 10. A compound of
formula V can be converted to a compound of formula LV by
protection of the hydroxyl groups with suitable protecting groups
such as t-butyldimethylsilyl. A compound of formula LVI can be
prepared from a compound of formula LV by ozonolysis. Treatment of
a compound of formula LVI with an amine and a reducing agent such
as sodium triacetoxyboro-hydride provides a compound of formula
LVII. Removal of the protecting groups from a compound of formula
LVII, with for example hydrogen fluoride, provides a compound of
formula V where X is oxygen, W is NR.sub.15 or oxygen, and G is
##STR00020##
##STR00021##
A compound of formula V where W is NR.sub.15, X is oxygen, and G
is
##STR00022## can be prepared as outlined in Scheme 11. A compound
of formula LVIII can be prepared from a compound of formula XXX by
treatment with an amine and standard amide bond coupling agents
such as EDCI and HOBT. A compound of formula LX can be prepared
from a compound of formula LVIII by N-deprotection, using for
example trifluoroacetic acid when P.sub.2 is a t-butyloxycarbonyl
group, followed by coupling of compounds of formula LIX and XIII
using standard amide bond coupling agents such as EDCI and HOBT. A
compound of formula LXI can be prepared from a compound of formula
LX by ring-closing metathesis. A compound of formula V can be
prepared from a compound of formula LXI following methods described
in Scheme 1.
##STR00023##
A compound of formula V where W is oxygen, X is oxygen, and G
is
##STR00024## can be prepared as outlined in Scheme 12. A compound
of formula LXII can be prepared from allylglycine by treatment with
nitrous acid. A compound of formula LXIII can be prepared from a
compound of formula LXII by treatment with an amine and standard
amide bond coupling agents such as EDCI and HOBT. A compound of
formula LXIV can be prepared from compounds of formula LXIII and
XIII using standard amide bond coupling agents such as EDCI and
HOBT. A compound of formula LXV can be prepared from a compound of
formula LXIV by ring-closing metathesis. A compound of formula V
can be prepared from a compound of formula LXV following methods
described in Scheme 1.
##STR00025##
Compounds of formula V where G is a 1,2-disubstituted ethyl group
can be prepared from a compound of formula V where G is a
1,2-disubstituted ethylene group by hydrogenation with a catalyst
such as palladium on carbon, as shown in Scheme 13. Furthermore,
compounds of formula V where G is a 1,2-disubstituted cyclopropyl
group can be prepared from a compound of formula V where G is a
1,2-disubstituted ethylene group by cyclopropanation with
diiodomethane and zinc-copper couple, as shown in Scheme 13.
##STR00026##
A compound of formula V where Z.sub.1 is oxygen can be prepared as
shown in Scheme 14. A compound of formula LXVII can be prepared
from an alpha-hydroxy ester LXVI and a
3-buten-1-yl-trifluoromethanesulfonate (or with a 3-butenyl bromide
and silver triflate). A compound of formula LXVII can be reduced
with a reducing agent such as diisobutylaluminum hydride to provide
a compound of formula LXVIII. Alternatively, a compound of formula
LXVIII can be obtained from a compound of formula LXVII by a two
step procedure involving reduction with lithium borohydride and
oxidation with pyridinium chlorochromate. This compound of formula
LXVIII can be substituted for a compound of formula XIV in Scheme 1
to give a compound of formula LXIX. Further elaboration of LXIX as
described above provides a compound of formula V where Z.sub.1 is
oxygen.
##STR00027##
Similarly, a compound of formula V where Z.sub.1 is NR.sub.23 can
be prepared as shown in Scheme 15. A compound of formula LXXI can
be prepared from an alpha-amino ester LXX and a
3-buten-1-yl-bromide. A compound of formula LXXI can be reduced
with a reducing agent such as diisobutylaluminum hydride to provide
a compound of formula LXXII. Alternatively, a compound of formula
LXXII can be obtained from a compound of formula LXXI by a two step
procedure involving reduction with lithium borohydride and
oxidation with pyridinium chlorochromate. This compound of formula
LXXII can be substituted for a compound of formula XIV in Scheme 1
to give a compound of formula LXXIII. Further elaboration of LXXIII
as described above provides a compound of formula V where Z.sub.1
is NR.sub.23.
##STR00028##
A compound of formula V where Z.sub.2 is oxygen can be prepared as
shown in Scheme 16. A compound of formula LXXV can be prepared from
a beta-hydroxy ester LXXIV and an allylating agent such as
allylbromide (or an allyl bromide and silver triflate). A compound
of formula LXXV can be reduced with a reducing agent such as
diisobutylaluminum hydride to provide a compound of formula LXXVI.
Alternatively, a compound of formula LXXVI can be obtained from a
compound of formula LXXV by a two step procedure involving
reduction with lithium borohydride and oxidation with pyridinium
chlorochromate. This compound of formula LXXVI can be substituted
for a compound of formula XIV in Scheme 1 to give a compound of
formula LXXVII. Further elaboration of LXXVII as described above
provides a compound of formula V where Z.sub.2 is oxygen.
##STR00029##
Similarly, a compound of formula V where Z.sub.2 is NR.sub.23 can
be prepared as shown in Scheme 17. A compound of formula LXXIX can
be prepared from a beta-amino ester LXXVIII and an allylating agent
such as allylbromide. A compound of formula LXXIX can be reduced
with a reducing agent such as diisobutylaluminum hydride to provide
a compound of formula .[.LIXXX.]. .Iadd.LXXX.Iaddend..
Alternatively, a compound of formula LXXX can be obtained from a
compound of formula LXXIX by a two step procedure involving
reduction with lithium borohydride and oxidation with pyridinium
chlorochromate. This compound of formula LXXX can be substituted
for a compound of formula XIV in Scheme 1 to give a compound of
formula LXXXI. Further elaboration of LXXXI as described above
provides a compound of formula V where Z.sub.2 is NR.sub.23.
##STR00030##
A compound of formula V where W is oxygen or NR.sub.15and X is H,H
can be prepared as shown in Scheme 18. A compound of formula V can
be converted to a compound of formula LXXXII, where P.sub.4 and
P.sub.5 are hydroxyl protecting groups, by treatment with a reagent
such as t-butyldimethylsilyltriflate. A compound of formula LXXXIII
can be prepared from a compound of formula LXXXII by treatment with
Lawesson's reagent. A compound of formula LXXXIV can be prepared
from a compound of formula LXXXIII by using a reducing agent such
as tri-n-butyltin hydride when W is oxygen or by treatment with
methyl iodide and sodium borohydride when W is NR.sub.15. Removal
of the protecting groups from a compound of formula LXXXIV, using
for example hydrogen fluoride when P.sub.4 and P.sub.5 are silyl
groups, provides a compound of formula V where W is oxygen or
NR.sub.15 and X is H,H.
##STR00031## ##STR00032##
A compound of formula V where W, X and Y are oxygen, and R.sub.1 is
alkyl or substituted alkyl can be prepared as shown in Scheme 19. A
compound of formula V can be protected to give a compound of
formula .[.LXXXV.]. .Iadd.LXXXXV.Iaddend., where P.sub.5 and
P.sub.6 are hydroxyl protecting groups, by treatment with a reagent
such as t-butyldimethylsilyl trifluoromethanesulfonate. A compound
of formula .[.LXXXVI.]. .Iadd.LXXXXVI .Iaddend.can be prepared from
a compound of formula .[.LXXXV.]. .Iadd.LXXXXV .Iaddend.by
treatment with a reducing agent such as sodium borohydride. A
compound of formula .[.LXXXVII.]. .Iadd.LXXXXVII .Iaddend.can be
prepared from a compound of formula .[.LXXXVI.]. .Iadd.LXXXXVI
.Iaddend.by protection of the hydroxyl group, where P.sub.7 is for
example p-methoxybenzyl, using p-methoxybenzyl
trichloroacetimidate. Removal of the protecting groups P.sub.5 and
P.sub.6 of a compound of formula LXXXXVII using, for example,
hydrogen fluoride in pyridine when P.sub.5 and P.sub.6 are
t-butyldimethylsilyl groups provides a compound of formula
LXXXXVIII which then can be selectively protected using for example
t-butyldimethylsilyl chloride to give a compound of formula LXXXXIX
where P.sub.8 is a t-butyldimethylsilyl group. A compound of
formula C can be prepared from a compound of formula LXXXXIX by
treatment with a base such as lithium diisopropylamide followed by
treatment with an alkylating agent such as methyl iodide. A
compound of formula C can be protected to give a compound of
formula CI, where P.sub.9 is a hydroxyl protecting group, by
treatment with a reagent such as t-butyldimethylsilyl
trifluoromethanesulfonate. A compound of formula CII can be
prepared from a compound of formula CI by removal of the P.sub.7
group using, for example, DDQ when P.sub.7 is a p-methoxybenzyl
group. A compound of formula V, where W, X and Y are oxygen, and
R.sub.1 is alkyl or substituted alkyl, can be prepared from a
compound of formula CII by oxidation using, for example, TPAP/NMO
followed by removal of the protecting groups using, for example,
hydrogen fluoride when P.sub.8 and P.sub.9 are silyl groups. This
compound of formula V can be further oxidized with
dimethyldioxirane as shown in Scheme 1 to provide the corresponding
epoxide compound of formula V.
##STR00033##
A compound of formula V where X is oxygen and Q is an olefin can be
prepared from a compound of formula V where X is oxygen and Q is an
oxirane ring by treatment with a reactive metallocene such as
titanocene, zirconocene or niobocene as shown in Scheme 20 (see for
example R. Schobert and U. Hohlein, Synlett, 465 466 (1990)).
##STR00034##
A compound of formula V where X is oxygen and W is NR.sub.15, where
R.sub.15 is hydrogen, can be prepared from a compound of formula V
where both X and W are oxygen as shown in Scheme 21. A compound of
formula CIII can be prepared from a compound of formula V where
both X and W are oxygen by formation of a pi-allylpalladium complex
using, for example, palladium tetrakistriphenylphosphine followed
by treatment with sodium azide (see, for example: Murahashi, S.-I.
et al., J. Org. Chem., 54:3292 (1989)). Subsequent reduction of a
compound of formula CIII with a reducing agent such as
triphenylphosphine provides a compound of formula CIV. A compound
of formula V where X is oxygen and W is NR.sub.15, where R.sub.15
is hydrogen, can be prepared from a compound of formula CIV by
macrolactamization using, for example, diphenylphosphoryl azide or
bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP).
##STR00035##
A compound of formula V where X is oxygen and W is NR.sub.15, where
R.sub.15 is alkyl, substituted alkyl, aryl, heteroaryl, cycloalkyl,
heterocyclo, O-alkyl, O-substituted alkyl, can be prepared from a
compound of formula V where both X and W are oxygen as shown in
Scheme 22. A compound of formula CV can be prepared from a compound
of formula V where both X and W are oxygen by formation of a
pi-allylpalladium complex using, for example, palladium
tetrakistriphenylphosphine followed by treatment with a primary
amine. A compound of formula V where X is oxygen and W is NR.sub.15
where R.sub.15 is alkyl, substituted alkyl, aryl, heteroaryl,
cycloalkyl, heterocyclo, OH, O-alkyl, O-substituted alkyl, can be
prepared from a compound of formula CV by macrolactamization using,
for example, diphenylphosphoryl azide or
bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP). In the
case where R.sub.15 is OH, it may be necessary to remove a
protecting group such as t-butyldimethylsilyl from an intermediate
where R.sub.15 is O-t-butyldimethylsilyl.
The in vitro assessment of biological activity of the compounds of
Formula V was performed as follows:
In Vitro Tubulin Polymerization
Twice cycled (2.times.) calf brain tubulin was prepared following
the procedure of Williams and Lee (see Williams, R. C., Jr. and
Lee, J. C., "Preparation of tubulin from brain.", Methods in
Enzymology, 85, Pt. D, 376 385 (1982)) and stored in liquid
nitrogen before use. Quantification of tubulin polymerization
potency is accomplished following a modified procedure of Swindell
et al. (see Swindell, C. S., Krauss, N. E., Horwitz, S. B. and
Ringel, I., "Biologically active taxol analogues with deleted
A-ring side chain substituents and variable C-2' configurations.",
J. Med Chem., 34:1176 1184 (1991)). These modifications, in part,
result in the expression of tubulin polymerization potency as an
effective concentration for any given compound. For this method,
different concentrations of compound in polymerization buffer (0.1M
MES, 1 mM EGTA, 0.5 mM MgCl.sub.2, pH 6.6) are added to tubulin in
polymerization buffer at 37.degree. in microcuvette wells of a
Beckman (Beckman Instruments) Model DU 7400 TN spectrophotometer. A
final microtubule protein concentration of 1.0 mg/ml and compound
concentration of generally 2.5, 5.0, and 10 .mu.M are used. Initial
slopes of OD change measured every 10 seconds were calculated by
the program accompanying the instrument after initial and final
times of the linear region encompassing at least 3 time points were
manually defined. Under these conditions linear variances were
generally <10.sup.-6, slopes ranged from 0.03 to 0.002
absorbance unit/minute, and maximum absorbance was 0.15 absorbance
units. Effective concentration (EC.sub.0.01) is defined as the
interpolated concentration capable of inducing an initial slope of
0.01 OD/minute rate and is calculated using the formula:
EC.sub.0.01=concentration/slope. EC.sub.0.01 values are expressed
as the mean with standard deviation obtained from 3 different
concentrations. EC.sub.0.01 values for the compounds in this
invention fall in the range 0.01 1000 .mu.M.
Cytoxicity (In-Vitro)
Cytoxicity was assessed in HCT-116 human colon carcinoma cells by
MTS
(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphenyl)--
2H-tetrazolium, inner salt) assay as reported in T. L. Riss et al.,
"Comparison of MTT, XTT, and a novel tetrazolium compound MTS for
in vitro proliferation and chemosensitivity assays.", Mol. Biol.
Cell, 3 (Suppl.):184a (1992). Cells were plated at 4,000 cell/well
in 96 well microtiter plates and 24 hours later drugs were added
and serial diluted. The cells were incubated at 37.degree. form 72
hours at which time the tetrazolium dye, MTS at 333 .mu.g/ml (final
concentration), in combination with the electron coupling agent
phenazine methosulfate at 25 .mu.M (final concentration) was added.
A dehydrogenase enzyme in live cells reduces the MTS to a form that
absorbs light at 492 nM which can be quantitated
spectrophotometrically. The greater the absorbance the greater the
number of live cells. The results are expressed as an IC.sub.50,
which is the drug concentration required to inhibit cell
proliferation (i.e. absorbance at 450 nM) to 50% of that of
untreated control cells. The IC.sub.50 values for compounds of this
invention fall in the range 0.01 1000 nM.
EXAMPLES
The following Examples illustrate the present invention.
Example 1
##STR00036##
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-Dihydroxy-5,5,7,9-tetramethyl-16-[1-met-
hyl-2-(2-methyl-4-thiazolyl)ethenyl]-1-aza-13(E)-cyclohexadecene-2,6-dione
A. N-[(2-Methyl)-1-propenyl]morpholine
To stirring morpholine (165.5 g, 1.9 mol) was added
isobutyraldehyde (173 mL, 1.9 mol) at a rate which did not allow
the temperature of the reaction to exceed 30.degree. C. After
complete addition, the reaction mixture was stirred at room
temperature for 2 h, and then the flask was equipped with a
Dean-Stark trap and heated at 160.degree. C. for 20 h. The reaction
mixture was then cooled to room temperature, and the flask was
equipped with a vigreux column distillation apparatus. Distillation
under high vacuum gave 135 g (50%) of Compound A as a clear
colorless oil. MS (M+H, 142).
B. 2,2-Dimethyl-3-oxopentanal
To a stirring solution of propionyl chloride (44 mL, 0.50 mol) in
ether (135 mL) at 0.degree. C. under nitrogen was added a solution
of Compound A (69 g, 0.50 mol) in ether (135 mL) over 45 min. After
addition was complete, the reaction mixture was stirred at reflux
for 2 h, and then stirred at room temperature for 16 h. The
reaction mixture was filtered, and the filter cake was washed with
ether (50 mL). The volatiles were removed in vacuo. The residue was
taken into H.sub.2O (80 mL) and the solution was adjusted to a pH
of 4. Ether was added (80 mL) and the biphasic mixture was stirred
for 16 h. The reaction mixture was poured into a separatory funnel,
the layers separated, and the aqueous layer was extracted with
ether (5.times.100 mL). The combined organics were dried
(MgSO.sub.4), filtered, and evaporated in vacuo. The residue was
distilled under high vacuum to give 10.4 g (16%) of Compound B as a
clear, colorless oil. MS (M-H, 127).
C. 4-tert-Butyldimethylsilyloxy-5,5-dimethyl-6-oxo-1-octene
To a solution of (-)-B-methoxydiisopinocamphenylborane (25.7 g, 81
mmol) in ether (80 mL) at 0.degree. C. under nitrogen was added 1.0
M allylmagnesium bromide in ether (77 mL, 77 mmol) over 1.5 h. The
reaction mixture was stirred at 25.degree. C. for 1 h, and then
concentrated in vacuo. The residue was extracted with pentane
(2.times.150 mL), and the extracts were filtered through Celite
under nitrogen. The combined extracts were then evaporated in vacuo
to give the B-allyldiisopinocamphenylborane. This material was
taken up in ether (200 mL) and cooled to -100.degree. C. under
nitrogen. A solution of Compound B (11.42 g, 89 mmol) in ether (90
mL) at -78.degree. C. was then added over a 1 h period. The
reaction mixture was stirred for an additional 0.5 h and methanol
(1.5 mL) was added. The reaction mixture was brought to room
temperature, treated with 3 N NaOH (32 mL) and 30% H.sub.2O.sub.2
(64 mL), and then kept at reflux for 2 h. The reaction mixture was
cooled to room temperature, the layers were separated, and the
organic phase was washed with H.sub.2O (500 mL). The combined
aqueous washes were re-extracted with ether (2.times.100 mL). The
combined organic extracts were washed with saturated aqueous NaCl
(100 mL), dried (MgSO.sub.4), filtered, and concentrated in vacuo.
This residue was taken up in CH.sub.2Cl.sub.2 (250 mL), cooled to
0.degree. C., and diisopropylethylamine (93 mL, 535 mmol) was
added. To the stirring solution was then added
tert-butyldimethylsilyl trifluoromethanesulfonate (69 g, 260 mmol)
slowly as to not increase the temperature above 10.degree. C. After
complete addition, the reaction mixture was poured into H.sub.2O
(650 mL), the layers were separated, and the aqueous layer was
extracted with CH.sub.2Cl.sub.2 (2.times.650 mL). The combined
organics were dried (Na.sub.2SO.sub.4), filtered, and concentrated
in vacuo. The residue was purified by flash chromatography eluting
with hexanes followed by 10% EtOAc/hexanes to give 17.2 g (78%) of
Compound C as a clear, colorless oil. The enantiomeric excess was
found to be 94% determined by .sup.1H NMR analysis of the Mosher's
ester of the alcohol. .sup.13C NMR (CDCl.sub.3, 80 MHz) d 215.8,
136.1, 116.5, 52.8, 39.0, 31.9, 26.0, 22.4, 20.1, 18.1, 7.6, -3.6,
-4.4.
D. 3-tert-Butyldimethylsiloxy-4,4-dimethyl-5-oxoheptanal
Through a solution of Compound C (10.8 g, 38.0 mmol) in
CH.sub.2Cl.sub.2 at -78.degree. C. was bubbled O.sub.3 until the
solution remained blue (1 h). O.sub.2 was then bubbled through for
15 min followed by N.sub.2 for 30 min after which time the solution
became clear. Triphenylphosphine (10 g, 38 mmol) was then added and
the reaction mixture was warmed to -35.degree. C. and stored for 16
h. The volatiles were removed in vacuo and the residue was purified
by flash chromatography eluting with 8% EtOAc/hexanes to give 8.9 g
(74%) of Compound D as a clear, colorless oil. .sup.1H NMR
(CDCl.sub.3, 300 MHz) d 9.75 (m, 1H), 4.53 (t, J=4.8 Hz, 1H), 3.40
3.60 (m, 4H), 1.10 (s, 3H), 1.07 (s, 3H), 0.98 (t, J=7.0 Hz, 3H),
0.83 (s, 9H), 0.07 (s, 3H), 0.04 (s, 3H).
E. 3-tert-Butyldimethylsiloxy-4,4-dimethyl-5-oxoheptanoic acid
To a solution of Compound D (3.90 g, 13.6 mmol) in t-butanol (75
mL) was added 2-methyl-2-butene (5.85 mL, 55.2 mmol), and then a
solution of sodium chlorite (4.61 g, 40.8 mmol) and sodium
phosphate monobasic (2.81 g, 20.4 mmol) in H.sub.2O (15 mL) was
added dropwise at room temperature. The reaction mixture was
stirred for 0.5 h and then the solvents were removed in vacuo. To
the residue was added H.sub.2O (150 mL) followed by extraction with
EtOAc (3.times.150 mL). The combined organic extracts were dried
(MgSO.sub.4), filtered, and the volatiles were removed in vacuo.
The residue was-purified by flash chromatography eluting with 20%
EtOAc/hexanes/1% AcOH to give 3.79 g (92%) of Compound E as a
clear, colorless, viscous oil. MS (M+H, 303).
F.
(R,R)--N-(2-Hydroxy-1-methyl-2-phenethyl)-N,2-(S)-dimethyl-6-hepteneami-
de
A suspension of LiCl (6.9 g, 0.16 mol) and preformed lithium
diisopropylamide (Aldrich, 2.0 M solution in
heptane/ethylbenzene/THF, 27.6 mL, 55 mmol) in additional THF (70
mL) at -78.degree. C. was treated dropwise with a solution of
(R,R)--N-(2-hydroxy-1-methyl-2-phenylethyl)-N-methyl propionamide
(6.0 g, 27 mmol, Meyers, A. G. et al., J. Am. Chem. Soc., 116:9361
(1994)) in THF (30 mL) over 10 min. The bright yellow, reaction
mixture was stirred at -78.degree. C. (1 h), at 0.degree. C. (15
min), and at 25.degree. C. (5 min) before being recooled to
0.degree. C. and treated with a solution of 5-bromo-1-pentene (4.8
mL, 40 mmol) in THF (5 mL). The reaction mixture was stirred at
0.degree. C. (24 h), poured into saturated aqueous NH.sub.4Cl (100
mL) and EtOAc (100 mL). The two phases were separated and the
aqueous phase was further extracted with EtOAc (3.times.100 mL).
The organic extracts were combined, washed with saturated aqueous
NaCl (200 mL), dried (Na.sub.2SO.sub.4), and concentrated in vacuo.
Flash chromatography (SiO.sub.2, 4.0.times.25 cm, 2%
MeOH--CHCl.sub.3) afforded Compound F (6.9 g, 88%) as a pale yellow
oil. MS (ESI.sup.+): 290 (M+H).sup.+; MS(ESI.sup.-): 288.2
(M-H).sup.-.
G. (S)-2-Methyl-6-heptenol
A 250 mL round-bottom flask at 0.degree. C. was charged
sequentially with pyrrolidine (2.6 mL, 30 mmol) and BH.sub.3-THF
complex (1.0 M in THF, 31 mL, 30 mmol). The borane-pyrrolidine
complex was warmed to 25.degree. C. (1 h), recooled to 0.degree.
C., and treated with n-butyllithium (1.6 M in hexane, 19 mL, 30
mmol) dropwise over 30 min while carefully maintaining an internal
temperature below 5.5.degree. C. The reaction mixture was stirred
at 0.degree. C. for an additional 30 min before a solution of
Compound F (3.0 g, 10 mmol) in THF (23 mL) was added dropwise over
10 min. The reaction mixture was stirred at 25.degree. C. (6 h)
before being quenched by the dropwise addition of aqueous 3 N HCl
(25 mL). The reaction mixture was then poured into aqueous 1 N HCl
(200 mL) and extracted with Et.sub.2O (4.times.80 mL). The combined
organics were washed with a 1:1 solution of saturated aqueous
NaCl--aqueous 1 N HCl (2.times.150 mL) and concentrated in vacuo.
An aqueous solution of NaOH (1 N, 200 mL) was added to the residue
and the suspension was stirred for 30 min. The mixture was
extracted with Et.sub.2O (3.times.100 mL) and the combined ether
layers were washed with a 1:1 solution of saturated aqueous
NaCl--aqueous 1 N NaOH (2.times.200 mL), dried (Na.sub.2SO.sub.4),
and concentrated in vacuo. Flash chromatography (SiO.sub.2,
4.0.times.25 cm, 15 25% Et.sub.2O-pentane gradient elution)
afforded Compound G (1.26 g, 95%) as a colorless oil.
[a].sup.25.sub.D-11 (c 12, CH.sub.2Cl.sub.2).
H. (S)-2-Methyl-6-heptenal
A solution of Compound G (0.24 g, 1.9 mmol) in CH.sub.2Cl.sub.2 (6
mL) was treated with pyridinium chlorochromate (0.61 g, 2.8 mmol)
and the reaction mixture was stirred at 25.degree. C. for 5 h. The
resulting dark brown viscous slurry was passed through a silica
gel-Celite plug (Celite 1.0.times.1 cm on top of SiO.sub.2,
1.0.times.5 cm, eluting with 50 mL of CH.sub.2Cl.sub.2). The
solvent was removed in vacuo to afford crude Compound H (0.15 g,
63%) as a colorless oil, which was sufficiently pure to use in
subsequent reactions. .sup.1H NMR (300 MHz, CD.sub.2Cl.sub.2) d
9.62 (s, 1H), 5.88 5.68 (m, 1H), 5.13 4.92 (m, 2H), 2.37 2.24 (m,
1H), 2.15 2.05 (m, 2H), 1.62 1.78 (m, 1H), 1.51 1.32 (m, 3H), 1.07
(d, 3H, J=7.0 Hz).
I.
(3S,6R,7S,8S)-3-tert-Butyldimethylsiloxy-4,4,6,8-tetramethyl-7-hydroxy--
5-oxo-12-tridecenoic acid
To a preformed LDA solution (Aldrich, 2.0 M solution in
heptane/ethylbenzene/THF, 3.8 mL, 7.6 mmol) in additional THF (25
mL) at -78.degree. C. was added a solution of Compound E (1.0 g,
3.4 mmol) in THF (5 mL) dropwise over 3 min. The reaction mixture
was stirred at -78.degree. C. (10 min), warmed to -40.degree. C.
(20 min), and recooled to -78.degree. C. before Compound H (0.56 g,
4.4 mmol) in THF (5 mL) was added. The reaction mixture was warmed
to -40.degree. C., stirred for 1 h, and diluted with saturated
aqueous NH.sub.4Cl (50 mL). The two layers were separated and the
aqueous phase was extracted with EtOAc (4.times.50 mL). The
combined organic layers were washed with saturated aqueous NaCl
(100 mL), dried (Na.sub.2SO.sub.4), and concentrated in vacuo.
Flash chromatography (SiO.sub.2, 2.5.times.20 cm, 2 5%
MeOH--CHCl.sub.3 gradient elution) followed by HPLC (YMC S-10, ODS,
30.times.500 mm column, eluting with MeOH at a flow rate of 20
mL/min) separation afforded the desired syn-aldol product Compound
I (0.60 g, 43%) and an undesired diastereomer (0.32 g, 22%) along
with starting Compound E (.about.10%). MS (ESI.sup.+): 879.3
(2M+Na).sup.+, 451.2 (M+Na).sup.+, 429.2 (M+H).sup.+;
MS(ESI.sup.-): 427.3 (M-H).sup.-. Stereochemistry was confirmed by
direct comparison of both the .sup.13C and .sup.1H NMRs of the
subsequent ester derivative (used in the synthesis of Epothilone C)
to the same intermediate previously described by K. C. Nicolaou et
al., Angew. Chem. Int. Ed. Engl., 36:166 (1997).
J. (S)-2-[N-[(tert-Butyloxy)carbonyl]amino]-4-pentenoic acid
A solution L-2-amino-4-pentenoic acid (NovaBiochem, 3.0 g, 26 mmol)
in THF--H.sub.2O (1:1, 200 mL) at 0.degree. C. was treated
sequentially with NaHCO.sub.3 (6.6 g, 78 mmol) and di-tert-butyl
dicarbonate (10.4 g, 1.8 mmol). The reaction mixture was warmed to
25.degree. C. and stirred for 16 h. The pH of the mixture was
adjusted to 4 by the careful addition of saturated aqueous citric
acid at 0.degree. C., and the mixture was extracted with EtOAc
(4.times.50 mL). The combined organic layers were washed with
saturated aqueous NaCl (75 mL), dried (Na.sub.2SO.sub.4), and
concentrated in vacuo. Flash chromatography (SiO.sub.2, 4.0.times.6
cm, 5 10% MeOH--CHCl.sub.3 gradient elution) afforded Compound J
(5.5 g, 99%) as a colorless oil. MS(ESI.sup.-): 429.3 (2M-H).sup.-,
214.1 (M-H).sup.-.
K.
(S)-2-[N.sup.2-[(tert-Butyloxy)carbonyl]amino]-N-methoxy-N-methyl-4-pen-
teneamide
A solution Compound J (2.9 g, 13 mmol) in CHCl.sub.3 (55 mL) at
0.degree. C. was treated sequentially with
N,O-dimethylhydroxylamine hydrochloride (1.4 g, 15 mmol),
1-hydroxybenzotriazole (2.0 g, 15 mmol), 4-methylmorpholine (4.4
mL, 40 mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (3.4 g, 18 mmol). The reaction mixture was gradually
warmed to 25.degree. C., stirred for 16 h, and diluted with
H.sub.2O (100 mL). The two layers were separated and the aqueous
phase was extracted with EtOAc (3.times.75 mL). The combined
organic phases were washed with aqueous 5% HCl (100 mL), saturated
aqueous NaHCO.sub.3 (100 mL), saturated aqueous NaCl (100 mL),
dried (Na.sub.2SO.sub.4), and concentrated in vacuo. Flash
chromatography (SiO.sub.2, 3.0.times.20 cm, 25 50% EtOAc-hexane
gradient elution) afforded Compound K (2.5 g, 71%) as a colorless
oil. MS (ESI.sup.+): 258.9 (M+H).sup.+, 202.9 (M-isobutylene) 158.9
(M-BOC); MS(ESI.sup.-): 257.2 (M-H).sup.-.
L. (S)-3-[N-[(tert-Butyloxy)carbonyl]amino]-5-hexen-2-one
A solution of Compound K (2.5 g, 1.0 mmol) in THF (65 mL) at
0.degree. C. was treated with methylmagnesium bromide (3.0 M in
Et.sub.2O, 8.1 mL, 2.4 mmol). The reaction mixture was stirred at
0.degree. C. (2.5 h) and carefully poured into saturated aqueous
NH.sub.4Cl (100 mL). The two layers were separated and the aqueous
phase was extracted with EtOAc (3.times.75 mL). The combined
organic extracts were washed with saturated aqueous NH.sub.4Cl (75
mL), H.sub.2O (75 mL), saturated aqueous NaCl (75 mL), dried
(MgSO.sub.4), and concentrated in vacuo. Flash chromatography
(SiO.sub.2, 3.0.times.20 cm, 10 25% EtOAc-hexane gradient elution)
afforded (S)-2-[N-[(tert-Butyloxy)carbonyl]amino]-5-hexene-2-one
(2.2 g, 67%) as a colorless oil. MS (ESI.sup.+): 213.9 (M+H).sup.+,
157.9 (M-isobutylene), 113.9 (M-BOC); MS(ESI.sup.-): 212.2
(M-H.sup.-).
M.
(S)-4-[3-[N-[(tert-Butyloxy)carbonyl]amino]-2-methyl-1(E),5-hexadienyl]-
-2-methylthiazole
A solution of 2-methyl-4-thiazolylmethyl diphenylphosphine oxide
(2.5 g, 8.0 mmol, Danishefsky et al., J. Org. Chem., 61:7998
(1996)) in THF (38 mL) at -78.degree. C. was treated with
n-butyllithium (1.6 M in hexane, 5.2 mL, 8.4 mmol) dropwise over 5
min. The resulting brilliant orange mixture was stirred for 15 min
at -78.degree. C., and treated with a solution of Compound L (0.81
g, 3.8 mmol) in THF (5 mL). After 10 min at -78.degree. C., the
cooling bath was removed and the reaction mixture was allowed to
warm to 25.degree. C. (2 h). The mixture was poured into saturated
aqueous NH.sub.4Cl (50 mL) and the two layers were separated. The
aqueous phase was extracted with Et.sub.2O (3.times.50 mL) and the
combined organic extracts were washed successively with H.sub.2O
(75 mL), saturated aqueous NaHCO.sub.3 (75 mL), saturated aqueous
NaCl (75 mL), dried (Na.sub.2SO.sub.4), and concentrated in vacuo.
Flash chromatography (SiO.sub.2, 4.0.times.30 cm, 10 20%
EtOAc-hexane gradient elution) afforded Compound M (0.23 g, 18%) as
a colorless oil along with recovered starting ketone (20 30%). MS
(ESI.sup.+): 309.1 (M+H).sup.+, 253.0 (M-isobutylene);
MS(ESI.sup.-): 307.3 (M-H).sup.-.
N. (S)-4-(3-Amino-2-methyl-1(E),5-hexadienyl-2-methylthiazole
Compound M (0.15 g, 0.49 mmol) was treated with 4.0 N HCl in
1,4-dioxane (5 mL) at 0.degree. C. (30 min) under Ar. The volatiles
were removed in vacuo, and the resulting white foam was dissolved
in cold saturated aqueous NaHCO.sub.3 (3 mL). The solution was
extracted with EtOAc (4.times.10 mL), and the combined EtOAc layers
were dried (Na.sub.2SO.sub.4) and concentrated in vacuo. Flash
chromatography (SiO.sub.2, 1.0.times.5 cm, 5 10% MeOH--CHCl.sub.3
gradient elution) afforded Compound N (88 mg, 88%) as a colorless
oil. MS (ESI.sup.+): 209.0 (M+H).sup.+; MS(ESI.sup.-): 207.2
(M-H).sup.-.
O.
(3S,6R,7S,8S)--N--(S)-[1-(2-Methyl-4-thiazolyl-2-methyl-1(E),5-hexadien-
-3-yl]-3-tert-butyldimethylsiloxy-4,4,6,8-tetramethyl-7-hydroxy-5-oxo-12-t-
rideceneamide
A solution of Compound N (88 mg, 0.42 mmol) in DMF (1.3 mL) at
0.degree. C. was treated sequentially with Compound I (0.15 g, 0.35
mmol), 1-hydroxybenzotriazole (49 mg, 0.36 mmol), and
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.10
g, 0.52 mmol). The reaction mixture was gradually warmed to
25.degree. C., stirred for 15 h, and diluted with H.sub.2O (3 mL).
The mixture was extracted with EtOAc (3.times.10 mL), and the
combined organic phases were washed with aqueous 5% HCl (10 mL),
saturated aqueous NaHCO.sub.3 (10 mL), and saturated aqueous NaCl
(10 mL), dried (Na.sub.2SO.sub.4), and concentrated in vacuo. Flash
chromatography (SiO.sub.2, 1.5.times.20 cm, 2.5% MeOH--CHCl.sub.3)
afforded Compound O (0.17 g, 77%) as a white foam. MS (ESI.sup.+):
619.3 (M+H).sup.+.
P.
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4-tert-Butyldimethylsiloxy-8-hydroxy-5,5-
,7,9-tetramethyl-16-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-1-aza-13(E)-
-cyclohexadecene-2,6-dione
A solution of Compound O (17 mg, 27 mmol) in degassed benzene (8.0
mL) was treated with Grubb's catalyst
[bis(tricyclohexyl-phosphine)benzylidine-ruthenium dichloride,
Strem Chemicals] (11 mg, 14 mmol) under Ar. The reaction mixture
was stirred at 25.degree. C. for 15 h and treated again with an
additional portion of catalyst (5.0 mg, 4.5 mmol). After 7
additional hours, the benzene was removed in vacuo, and the black
viscous residue was passed through a pad of silica gel (1.0.times.3
cm) eluting with Et.sub.2O (25 mL). The eluent was concentrated in
vacuo to afford a separable 5:1 (E/Z) mixture of geometric isomers.
PTLC (SiO.sub.2, 1 mm plate, 2 elutions with a 1:1:1 solution of
hexane-toluene-ethyl acetate) afforded the E-isomer Compound P (5.1
mg, 34%) and the corresponding Z-isomer (1.0 mg, 6.7%). For
Compound P: MS (ESI.sup.+): 1181.7 (2M+H).sup.+, 591.4 (M+H).sup.+.
For the Z-isomer: MS (ESI.sup.+): 1181.5 (2M+H).sup.+, 613.2
(M+Na).sup.+, 591.2 (M+H).sup.+; MS (ESI.sup.-): 589.3
(M-H).sup.-.
Q.
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-Dihydroxy-5,5,7,9-tetramethyl-16-[1--
methyl-2-(2-methyl-4-thiazolyl)ethenyl]-1-aza-13(E)-cyclohexadecene-2,6-di-
one
To a 1 dram vial charged with Compound P (2.3 mg, 3.9 mmol) in
CH.sub.2Cl.sub.2 (0.4 mL) at 0.degree. C. was added trifluoroacetic
acid (0.1 mL). The reaction mixture was sealed under a blanket of
Ar and stirred at 0.degree. C. After 4 h, the volatiles were
removed under a constant stream of Ar at 0.degree. C. Saturated
aqueous NaHCO.sub.3 (1 mL) and EtOAc (1 mL) were added to the
residue and the two layers were separated. The aqueous phase was
extracted with EtOAc (4.times.1 mL), and the combined EtOAc layers
were dried (Na.sub.2SO.sub.4) and concentrated in vacuo. PTLC
(SiO.sub.2, 20.times.10.times.0.025 cm, eluting with 5%
MeOH--CHCl.sub.3) afforded
[4S-[4R*,7S*,8S*,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9-tetramethyl-16-[1-me-
thyl-2-(2-methyl-4-thiazolyl)ethenyl]-1-aza-13(E)-cyclohexadecene-2,6-dion-
e (1.3 mg, 68%) as a white film. MS (ESI.sup.+): 953.5
(2M+H).sup.+, 477.3 (M+H).sup.+; MS (ESI.sup.-): 475.5
(M-H).sup.-.
Example 2
The following compounds can be made following the reaction schemes
previously disclosed:
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12,16-pen-
tamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,13,17-trioxabicycl-
o[14.1.0]heptadecane-5,9-dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12-tetram-
ethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,13,17-trioxabicyclo[1-
4.1.0]heptadecane-5,9-dione;
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9,13-pentamethyl-16-[1-
-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-1,10-dioxa-13-cyclohexadecene-2,6-
-dione; [4S-[4R*,7S*
,8R*,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9-tetramethyl-16-[1-methyl-2-(2-me-
thyl-4-thiazolyl)ethenyl]-1,10-dioxa-13-cyclohexadecene-2,6-dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12,16-pen-
tamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,14,17-trioxabicycl-
o[14.1.0]heptadecane-5,9-dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12-tetram-
ethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,14,17-trioxabicyclo[1-
4.1.0]heptadecane-5,9-dione; [4S-[4R* ,7S*,8R*
,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9,13-pentamethyl-16-[1-methyl-2-(2-met-
hyl-4-thiazolyl)ethenyl]-1,11-dioxa-13-cyclohexadecene-2,6-dione;
[4S-[4R*,7S*,8R*
,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9-tetramethyl-16-[1-methyl-2-(2-methyl-
-4-thizolyl)ethenyl]-1,11-dioxa-13-cyclohexadecene-2,6-dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12,16-pen-
tamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,17-dioxabicyclo[14-
.1.0]heptadecane-9-one;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12-tetram-
ethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,17-dioxabicyclo[14.1.-
0]heptadecane-9-one;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-3,8,8,10,12,16-h-
examethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,17-dioxabicyclo[1-
4.1.0]heptadecane-5,9-dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-3,8,8,10,12-pent-
amethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,17-dioxabicyclo[14.-
1.0]heptadecane-5,9-dione;
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9,13,16-hexamethyl-16--
[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-1-oxa-13-cyclohexadecene-2,6-di-
one;
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9,16-pentamethyl-1-
6-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-1-oxa-13-cyclohexadecene-2,6--
dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-6,8,8,10,1-
2,16-hexamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,17-dioxabic-
yclo[14.1.0]heptadecane-5,9-dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-6,8,8,10,12-pent-
amethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,17-dioxabicyclo[14.-
1.0]heptadecane-5,9-dione;
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9-tetramethyl-16-[1-(2-
-methyl-4-thiazolyl)ethenyl]-1-aza-13-cyclohexadecene-2,6-dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-4,8,8,10,12,16-h-
examethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-aza-17-oxabicyclo-
[14.1.0]heptadecane-5,9-dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-4,8,8,10,12-pent-
amethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-aza-17-oxabicyclo[1-
4.1.0]heptadecane-5,9-dione;
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-dihydroxy-1,5,5,7,9,13-hexamethyl-16-[-
1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-1-aza-13-cyclohexadecene-2,6-dio-
ne;
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-dihydroxy-1,5,5,7,9-pentamethyl-16--
[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-1-aza-13-cyclohexadecene-2,6-di-
one;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12,16-
-pentamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-13-aza-4,17-diox-
abicyclo[14.1.0]heptadecane-5,9-dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12-tetram-
ethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-13-aza-4,17-dioxabicycl-
o[14.1.0]heptadecane-5,9-dione;
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9,13-pentamethyl-16-[1-
-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-10-aza-1-oxa-13-cyclohexadecene-2-
,6-dione;
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9-tetramethyl-
-16-[1-(2-methyl-4-thiazolyl)ethenyl]-10-aza-1-oxa-13-cyclohexadecene-2,6--
dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12,-
16-pentamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-14-aza-4,17-di-
oxabicyclo[14.1.0]heptadecane-5,9-dione;
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12-tetram-
ethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-14-aza-4,17-dioxabicycl-
o[14.1.0]heptadecane-5,9-dione;
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9,13-pentamethyl-16-[1-
-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-11-aza-1-oxa-13-cyclohexadecene-2-
,6-dione;
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-dihydroxy-5,5,7,9-tetramethyl-
-16-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-11-aza-1-oxa-13-cyclohexade-
cene-2,6-dione;
[1S-[1R*,3R*,7R*,10S*,11R*,12R*,16S*]]-N-phenyl-7,11-dihydroxy-8,8,10,12,-
16-pentamethyl-5,9-dioxo-4,17-dioxabicyclo[14.1.0]heptadecane-3-carboxamid-
e;
[1S-[1R*,3R*,7R*,10S*,11R*,12R*,16S*]]-N-phenyl-7,11-dihydroxy-8,8,10,1-
2-tetramethyl-5,9-dioxo-4,17-dioxabicyclo[14.1.0]heptadecane-3-carboxamide-
;
[4S-[4R*,7S*,8R*,9R*,15R*]]-N-phenyl-4,8-dihydroxy-5,5,7,9,13-pentamethy-
l-2,6-dioxo-1-oxa-13-cyclohexadecene-16-carboxamide;
[4S-[4R*,7S*,8R*,9R*,15R*]]-N-phenyl-4,8-dihydroxy-5,5,7,9-tetramethyl-2,-
6-dioxo-1-oxa-13-cyclohexadecene-16-carboxamide;
[1S[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12,16-pent-
amethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)cyclopropyl]-4,17-dioxabicyclo-
[14.1.0]heptadecane-5,9-dione; and
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-dihydroxy-8,8,10,12-tetram-
ethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)cyclopropyl]-4,17-dioxabicyclo[1-
4.1.0]heptadecane-5,9-dione.
##STR00037##
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12,16-pent-
amethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-aza-17-oxabicyclo[1-
4.1.0]heptadecane-5,9-dione
A.
(3S,6R,7S,8S,12R,13S,15S)-15-Azido-12,13-epoxy-4,4,6,8,12,16-hexamethyl-
-7-hydroxy-17-(2-methyl-4-thiazolyl)-5-oxo-16-heptadecenoic
acid
A solution of epothilone B (0.35 g, 0.69 mmol) in degassed THF (4.5
mL) was treated with a catalytic amount (80 mg, 69 mmol) of
tetrakis(triphenylphosphine) palladium (0) and the suspension was
stirred at 25.degree. C., under Ar for 30 min. The resulting bright
yellow, homogeneous solution was treated all at once with a
solution of sodium azide (54 mg, 0.83 mmol) in degassed H.sub.2O
(2.2 mL). The reaction mixture was warmed to 45.degree. C. for 1 h,
diluted with H.sub.2O (5 mL) and extracted with EtOAc (4.times.7
mL). The organic extracts were washed with saturated aqueous NaCl
(15 mL), dried (Na.sub.2SO.sub.4), and concentrated in vacuo. The
residue was purified by flash chromatography (SiO.sub.2,
3.0.times.15 cm, 95:5.0:0.5 CHCl.sub.3--MeOH--AcOH) to afford
Compound A (0.23 g, 61%) as a colorless oil. MS (ESI.sup.+): 551
(M+H).sup.+; MS(ESI.sup.-): 549 (M-H).sup.-.
B.
(3S,6R,7S,8S,12R,13S,15S)-15-Amino-12,13-epoxy-4,4,6,8,12,16-hexamethyl-
-7-hydroxy-17-(2-methyl-4-thiazolyl)-5-oxo-16-heptadecenoic
acid
A solution of Compound A (0.23 g, 0.42 mmol) in THF (4.0 mL) was
treated with H.sub.2O (23 mL, 1.25 mmol) and polymer supported
triphenylphosphine (Aldrich, polystyrene cross-linked with 2% DBL.,
0.28 g, 0.84 mmol) at 25.degree. C. The resulting suspension was
stirred at 25.degree. C. under Ar (32 h), filtered through a Celite
pad and concentrated in vacuo. The residue was purified by flash
chromatography (SiO.sub.2, 1.5.times.10 cm, 95:5.0:0.5 to 90:10:1.0
CHCl.sub.3-MeOH--AcOH gradient elution) to afford Compound B (96
mg, 44%) as a colorless oil. MS (ESI.sup.+): 525.2 (M+H).sup.+;
MS(ESI.sup.-): 523.4 (M-H).sup.-.
Alternatively, to a 25 mL round-bottom flask charged with Compound
A (0.26 g, 0.47 mmol) and PtO.sub.2 (0.13 g, 50 wt %) was added
absolute EtOH under Ar. The resulting black mixture was stirred
under one atmosphere of H.sub.2 for 10 h, after which time the
system was purged with N.sub.2 and an additional portion of
PtO.sub.2 (65 mg, 25 wt %) was added. Once again the reaction
mixture was stirred under a blanket of H.sub.2 for 10 h. The system
was then purged with N.sub.2, and the reaction mixture was filtered
through a Celite pad eluting with CH.sub.2Cl.sub.2 (3.times.25 ML).
The solvents were removed in vacuo and the residue was purified as
described above to afford Compound B (0.19 g, 75%).
Alternatively, a solution of Compound A (20 mg, 36 mmol) in THF
(0.4 mL) was treated with triphenylphosphine (19 mg, 73 mmol) under
Ar. The reaction mixture was warmed to 45.degree. C., stirred for
14 h and cooled to 25.degree. C. The resulting iminophosphorane was
treated with ammonium hydroxide (28%, 0.1 mL) and once again the
reaction mixture was warmed to 45.degree. C. After 4 h, the
volatiles were removed in vacuo and the residue was purified as
described above to afford Compound B (13 mg, 70%).
C.
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12,16-p-
entamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-aza-17-oxabicycl-
o[14.1.0]heptadecane-5,9-dione
A solution of Compound B (0.33 g, 0.63 mmol) in degassed DMF (250
mL) was treated with solid NaHCO.sub.3 (0.42 g, 5.0 mmol) and
diphenylphosphoryl azide (0.54 mL, 2.5 mmol) at 0.degree. C. under
Ar. The resulting suspension was stirred at 4.degree. C. for 24 h,
diluted with phosphate buffer (250 mL, pH=7) at 0.degree. C. and
extracted with EtOAc (5.times.100 mL). The organic extracts were
washed with 10% aqueous LiCl (2.times.125 mL), dried
(Na.sub.2SO.sub.4) and concentrated in vacuo. The residue was first
purified by flash chromatography (SiO.sub.2, 2.0.times.10 cm, 2 5%
MeOH--CHCl.sub.3 gradient elution) and then repurified using a
Chromatotron (2 mm SiO.sub.2, GF rotor, 2 5% MeOH--CHCl.sub.3
gradient elution) to afford the title compound (0.13 g, 40%) as a
colorless oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 6.98 (s,
1H), 6.71 (d, 1H, NH, J=8.1 Hz), 6.56 (s, 1H), 4.69 4.62 (m, 1H),
4.18 4.12 (m, 1H), 4.01 3.96 (m, 1H), 3.86 (s, 1H), 3.38 3.34 (m,
1H), 2.82 (dd, 1H, J=5.6, 6.0Hz), 2.71 (s, 3H), 2.58 (s, 1H), 2.43
(dd, 1H, J=9.0, 14.5Hz), 3.34 (dd, 1H, J=3.0, 14.5Hz), 2.14 (s,
3H), 2.05 1.92 (m, 2H), 1.82 1.41 (a series of multiplets, 7H),
1.35 (s, 3H), 1.28 (s, 3H), 1.18 (d, 3H, J=6.8 Hz), 1.14 (s 3H),
1.00 (d, 3H, J=6.8 Hz); MS (ESI.sup.+): 507.2
(M+H).sup.+;.sup.-MS(ESI.sup.-): 505.4 (M-H).sup.-.
Example 4
Process for Reduction of Oxirane Ring of Epothilone and Epothilone
Analogs
To a two-necked flask was added chopped pieces of magnesium
turnings (24 mg, 1.0 mmol). The flask was flame-dried under vacuum
and cooled under argon. Bis(cyclopentadienyl)titanium dichloride
(250 mg, 1.0 mmol) was added followed by anhydrous THF (5 mL). The
stirring suspension was evacuated with low vacuum, and the reaction
flask was refilled with argon. The red suspension became dark,
turning a homogeneous deep green after 1.5 h with nearly all the
magnesium metal being consumed. An aliquot (3.5 mL, 0.70 mmol, 3.5
eq) was removed and cooled to -78.degree. C. under argon. To this
solution was added epothilone A (99 mg, 0.20 mmol, 1.0 eq). The
reaction mixture was warmed to room temperature and stirred for 15
min. The volatiles were removed in vacuo and the residue was
chromatographed two times on silica (25 g), eluting with 35%
EtOAc/hexanes to give 76 mg (80%) of epothilone C as a pale yellow
viscous oil.
Example 5
##STR00038##
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12-tetrame-
thyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-aza-17-oxabicyclo[14.1-
.0]heptadecane-5,9-dione
A.
(3S,6R,7S,8S,12R,13S,15S)-15-Azido-3,7-dihydroxy-12,13-epoxy-4,4,6,8,16-
-pentamethyl-17-(2-methyl-4-thiazolyl)-5-oxo-16(E)-heptadecenoic
acid
Tetrakis(triphenylphosphine)palladium(0) (1.17 g, 1.01 mmol, 0.10
eq) was added to a solution of epothilone A (4.97 g, 10.1 mmol, 1.0
eq) in degassed THF (100 ml) at room temperature and was stirred
for 30 minutes-under argon. Sodium-azide (0.980 g, 15.1 mmol, 1.5
eq) was added to the above reaction mixture followed by the
addition of degassed water (10 ml). The reaction mixture-was heated
to 45.degree. C. for one hour, cooled to room temperature, diluted
with ethyl acetate (300 ml) and further diluted with water (150
ml). The aqueous layer was extracted with ethyl acetate
(3.times.100 ml). The combined organic extracts were washed with
brine (150 ml), dried (sodium sulfate), filtered and concentrated
under vacuum. The oily residue was purified by flash silica gel
chromatography (eluting 0 5% methanol/chloroform with 0.1% of
acetic acid) to afford Compound A (1.84 g, 34.0% yield) as glassy
solid. MS (ESI.sup.+): 537 (M+H).sup.+; MS (ESI): 535
(M-H).sup.-.
B.
(3S,6R,7S,8S,12R,13S,15S)-15-Amino-3,7-dihydroxy-12,13-epoxy-4,4,6,8,16-
-pentamethyl-17-(2-methyl-4-thiazolyl)-5-oxo-16(E)-heptadecenoic
acid
Platinum oxide (0.980 g, 4.30 mmol, 1.25 eq) was added to a
solution of Compound A (1.85 g, 3.44 mmol, 1.0 eq) in absolute
ethanol (137 ml). The reaction mixture was stirred vigorously under
a hydrogen balloon for 16 hours at room temperature. The reaction
mixture was filtered and the filtrate was concentrated under
vacuum. The oily residue was purified by preparative HPLC (YMC S-15
ODS 50.times.500 mm column, 45 minutes/gradient, 0 100% B, 50
ml/min, retention time=17 minutes, A=0.1% acetic acid/5%
acetonitrile/95% water, B=0.1% acetic acid/5% water/95%
acetonitrile). The appropriate fractions were concentrated under
vacuum and the residue was lyophilized from aqueous acetonitrile to
afford Compound B (1.33 g, 76.0% yield) as a colorless solid. MS
(ESI.sup.+): 511(M+H).sup.+; MS (ESI.sup.-): 509 (M-H).sup.-.
C.
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12-tetr-
amethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-aza-17-oxabicyclo[1-
4.1.0]heptadecane-5,9-dione
Compound B (0.860 g, 1.68 mmol, 1.0 eq) was dissolved in anhydrous
DMF (0.00250M, 672 ml) and degassed for one hour at room
temperature. The solution was cooled to 0.degree. C., and anhydrous
sodium bicarbonate (1.13 g, 13.4 mmol, 4.0 eq) and
diphenylphosphoryl azide (1.85 g, 6.72 mmol, 8.0 eq) were added
under argon. The reaction mixture was kept at 4.degree. C. under
argon and stirred 16 hours. The reaction mixture was then cooled to
-60.degree. C., and pH 7 phosphate buffer (400 ml) was added slowly
to quench the reaction. The temperature was kept below -30.degree.
C. The above mixture was allowed to warm to room temperature slowly
and extracted with ethyl acetate (1 liter). The aqueous layer was
washed with ethyl acetate (4.times.300 ml). The organic extracts
were combined, washed with 10% LiCl (500 ml), dried (sodium
sulfate), filtered and concentrated under vacuum. The oily residue
was purified by preparative HPLC (YMC S-15 ODS 50.times.500 mm
column, 45 minutes/gradient, 0 100% B, 50 ml/min, retention time=35
minutes, A=5% acetonitrile/95% water, B=5% water/95% acetonitrile).
The appropriate fractions were concentrated under vacuum and the
residue was lyophilized from aqueous acetonitrile to afford title
compound (0.220 g, 26.0% yield) as a colorless solid. MS
(ESI.sup.+): 493 (M+H).sup.+, MS (ESI.sup.-): 491 (M-H).sup.-.
Example 6
##STR00039##
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-Dihydroxy-5,5,7,9,13-pentamethyl-16-[1--
methyl-2-(2-methyl-4-thiazolyl)ethenyl]-1-aza-13(Z)-cyclohexadecene-2,6-di-
one
Tungsten hexachloride (0.19 g, 0.49 mmol, 0.5 equiv) was dissolved
in THF (5.0 ml) and the solution was cooled to -78.degree. C.
n-Butyllithium in hexane (1.6M, 0.63 ml, 1.0 mmol, 1.0 equiv) was
added in one portion and the reaction mixture was allowed to warm
to room temperature over 20 minutes (the solution turned dark green
upon warming to rt). A 0.1M solution of the prepared tungsten
reagent (0.79 ml, 0.079 mmol, 2.0 equiv.) was added to Compound 4C
(0.020 g, 0.039 mmol, 1.0 equiv) at room temperature. The reaction
mixture was stirred at room temperature for 30 minutes and then was
quenched with saturated NaHCO.sub.3 (2.0 ml). The quenched solution
was diluted with water (10 ml) and the solution was extracted with
CH.sub.2Cl.sub.2 (4.times.20 ml). The combined organic extracts
were dried (Na.sub.2SO.sub.4), filtered and concentrated under
vacuum. The inorganics were removed by passing the residue through
a silica gel plug (eluting with 19/1 CHCl.sub.3/MeOH). The eluent
was concentrated under vacuum. The residue was purified by PHPLC
(YMC-S5 ODS, 30 100% B, A=5% aq CH.sub.3CN, B=95% aqueous
CH.sub.3CN, 3 ml/min., 220 nm., 30 min. gradient) and the
appropriate fractions were concentrated under vacuum. The sticky
solid was lyophilized from aqueous acetonitrile to afford the title
compound (4.3 mg, 29%) as a white solid. TLC: R.sub.f=0.57 (9/1
CHCl.sub.3/MeOH, visualization by UV); HRMS: (M+H).sup.+
calc=491.29436, found=491.2934.
Example 7
##STR00040##
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12,16-pent-
amethyl-3-[1-methyl-2-(2-hydroxymethyl-4-thiazolyl)ethenyl]-4-aza-17-oxabi-
cyclo[14.1.0]heptadecane-5,9-dione
##STR00041##
A.
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12,16-p-
entamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4,17-dioxabicyclo[-
14.1.0]heptadecane-5,9-dione,N-oxide
A solution of epothilone B (2.0 g, 3.9 mmol) in CH.sub.2Cl.sub.2
(30 mL) was treated with 3-chloroperoxybenzoic acid (1.0 g, 5.9
mmol) at 25.degree. C., under Ar for 2 h. An additional 0.5 g (3.0
mmol) of 3-chloroperoxybenzoic acid was added and the reaction
mixture was then stirred for 2 h. The reaction mixture was filtered
and the filtrate was concentrated in vacuo. The residue was
dissolved in EtOAc (100 mL), washed with saturated aqueous
NaHCO.sub.3 (75 mL), 5% aqueous Na.sub.2SO.sub.3 (75 mL), H.sub.2O
(75 mL), dried (Na.sub.2SO.sub.3) and concentrated in vacuo. The
residue was purified by flash chromatography (SiO.sub.2,
4.5.times.30 cm, 2 10% MeOH--CHCl.sub.3 gradient elution) to afford
Compound A (1.04 g, 50%) as a white solid. MS (ESI.sup.+): 524.3
(M+H).sup.+; MS (ESI.sup.-): 522.5 (M-H).sup.-.
##STR00042##
B. [1S-[1R*,3R*(E),7R*,10
S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12,16-pentamethyl-3-[1-methyl-2-
-(2-hydroxymethyl-4-thiazolyl)ethenyl]-4,17-dioxabicyclo[14.1.0]heptadecan-
e-5,9-dione[Epothilone F]
To a solution of Compound A (0.46 g, 0.88 mmol) in CH.sub.2Cl.sub.2
(10 mL) in a resealable tube was added 2,6-lutidine (0.82 mL, 7.0
mmol) and trifluoroacetic anhydride (0.87 mL, 6.2 mmol) under Ar.
The reaction vessel was sealed under Ar, heated to 75.degree. C.
(12 min), cooled to 25.degree. C., and the volatiles were removed
under a steady stream of N.sub.2. The reaction tube was then placed
on a high vacuum pump for 15 min. The resulting residue was
dissolved in MeOH (10 mL) and treated with ammonium hydroxide (28
30% NH.sub.4 in H.sub.2O, 1.0 mL). The mixture was heated to
45.degree. C. (10 min), and the volatiles were removed in vacuo.
The crude reaction mixture was purified by HPLC (YMC S-15 ODS
30.times.500 mm column, 50% acetonitrile-H.sub.2O isocratic
conditions, flow rate=20 mL/min, retention time=28 min). The
appropriate fractions were concentrated under vacuum and the
residue was lyophilized from aqueous acetonitrile to afford
Compound B (0.22 g, 48%) as a white solid. MS (ESI.sup.+): 524.3
(M+H).sup.+, 1047.6 (2M+H).sup.+; MS (ESI.sup.-): 522.5
(M-H).sup.-.
##STR00043##
C.
(3S,6R,7S,8S,12R,13S,15S)-15-Azido-3,7-Dihydroxy-12,13-epoxy-4,4,6,8,12-
,16-hexamethyl-17-(2-hydroxymethyl-4-thiazolyl)-5-oxo-16(E)-heptadecenoic
acid
A solution of Compound B (0.18 g, 0.34 mmol) in degassed THF (3.0
mL) was treated with a catalytic amount (40 mg, 3.4.times.10.sup.-2
mmol) of tetrakis(triphenylphosphine) palladium(0) and the
suspension was stirred at 25.degree. C., under Ar for 30 min. The
resulting bright yellow, homogeneous solution was treated all at
once with a solution of sodium azide (27 mg, 0.41 mmol) in degassed
H.sub.2O (1.5 mL). The reaction mixture was warmed to 45.degree. C.
for 1 h, diluted with H.sub.2O (5 mL) and extracted with EtOAc
(4.times.10 mL). The organic extracts were washed with saturated
aqueous NaCl (15 mL), dried (Na.sub.2SO.sub.4), and concentrated in
vacuo. The residue was purified by flash chromatography (SiO.sub.2,
2.5.times.15 cm, 95:5 CHCl.sub.3-MeOH to 95:5.0:0.5
CHCl.sub.3-MeOH--AcOH gradient elution) to afford Compound C (39
mg, 20%) as a colorless oil. MS (ESI.sup.+): 567.4 (M+H).sup.+,
1133.6 (2M+H).sup.+; MS (ESI.sup.-): 565.5 (M-H).sup.-, 1131.8
(2M-H).sup.-.
##STR00044##
D.
(3S,6R,7S,8S,12R,13S,15S)-15-Amino-3,7-dihydroxy-12,13-epoxy-4,4,6,8,12-
,16-hexamethyl-17-(2-hydroxymethyl-4-thiazolyl)-5-oxo-16(E)-heptadecenoic
acid
To a 10 mL round-bottom flask charged with Compound C (40 mg, 71
mmol) and PtO.sub.2 (12 mg, 30 wt %) was added absolute EtOH (3 mL)
under Ar. The resulting black mixture was stirred under one
atmosphere of H.sub.2 for 10 h. The system was then purged with
N.sub.2 and the reaction mixture was filtered through a nylon
membrane (washing with 25 mL of MeOH). The solvents were removed in
vacuo to afford Compound D (29 mg, 76%) as a foam, which was
sufficiently pure to use in the next step. LCMS: 541.3
(M+H).sup.+.
##STR00045##
E.
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12,16-p-
entamethyl-3-[1-methyl-2-(2-hydroxymethyl-4-thiazolyl)ethenyl]-4-aza-17-ox-
abicyclo[14.1.0]heptadecane-5,9-dione
A solution of Compound D (29 mg, 54 mmol) in degassed DMF (21 mL)
was treated with solid NaHCO.sub.3 (36 mg, 0.43 mmol) and
diphenylphosphoryl azide (46 mL, 0.21 mmol) at 0.degree. C. under
Ar. The resulting suspension was stirred at 4.degree. C. for 19 h,
cooled to -40.degree. C., diluted with 25 mL of pH 7 phosphate
buffer (carefully adding such that the internal temperature remains
below -30.degree. C.), and extracted with EtOAc (4.times.10 mL).
The organic extracts were washed with cold 10% aqueous LiCl (25
mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The
residue was purified using a chromatotron (1 mm SiO.sub.2 GF rotor,
2 5% MeOH--CHCl.sub.3 gradient elution) to afford the title
Compound E (9.1 mg, 34%) as a colorless oil. MS (ESI.sup.+): 523.2
(M+H).sup.+; MS (ESI.sup.-): 521.5 (M-H).sup.-.
Example 8
##STR00046##
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-Dihydroxy-5,5,7,9,13-pentamethyl-16-[1--
methyl-2-(2-hydroxymethyl-4-thiazolyl)ethenyl]-1-aza-13(Z)-cyclohexadecene-
-2,6-dione
##STR00047##
A.
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12,16-p-
entamethyl-3-[1-methyl-2-(2-tert-butyldiphenylsilyloxymethyl-4-thiazolyl)e-
thenyl]-4-aza-17-oxabicyclo[14.1.0]heptadecane-5,9-dione
A solution of Compound 7E (6.8 mg, 13 mmol) in CH.sub.2Cl.sub.2
(0.5 mL) was treated with triethylamine (2.7 mL, 20 mmol),
4-N,N-dimethylaminopyridine (0.2 mg, 1.3 mmol) and
tert-butyldiphenylsilyl chloride (3.7 mL, 14 mmol) at 0.degree. C.
under Ar. The reaction mixture was gradually warmed to 25.degree.
C. (1 h), cooled to 0.degree. C., quenched by the addition of
saturated aqueous NaHCO.sub.3 (1 mL), and extracted with EtOAc
(4.times.2 mL). The combined organic extracts were washed with
brine (5 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo.
The residue was purified by flash chromatography (SiO.sub.2,
1.0.times.5 cm, 2 5% MeOH--CHCl.sub.3 gradient elution) to afford
Compound A (7.0 mg, 71%) as a colorless oil. MS (ESI.sup.+): 761.5
(M+H).sup.+; MS (ESI.sup.-): 759.7 (M-H).sup.-.
##STR00048##
B.
[4S-[4R*,7S*,8R*,9R*,15R*(E)]]-4,8-Dihydroxy-5,5,7,9,13-pentamethyl-16--
[1-methyl-2-(2-hydroxymethyl-4-thiazolyl)ethenyl]-1-aza-13(Z)-cyclohexadec-
ene-2,6-dione
A solution of tungsten(IV) chloride (0.10 g, 0.25 mmol) in
anhydrous THF at -78.degree. C. was treated with n-BuLi (1.6 M in
hexanes, 0.32 mL, 0.50 mmol) under Ar. The reaction mixture was
warmed to 25.degree. C. over 40 min and then recooled to 0.degree.
C. An aliquot of the resulting deep-green, homogeneous solution
(0.2 mL, 20 mmol) was added to a 1 dram vial charged with Compound
A (7.0 mg, 9.2 mmol) at 0.degree. C. under Ar. The reaction mixture
was warmed to 25.degree. C., stirred for 30 min, quenched by the
addition of saturated aqueous NaHCO.sub.3 (0.5 mL) and extracted
with EtOAc (4.times.1 mL). The combined organic extracts were dried
(Na.sub.2SO.sub.4) and concentrated in vacuo. The residue was
purified by preparative TLC (SiO.sub.2, 20.times.20.times.0.025 cm,
eluting with 5% MeOH--CHCl.sub.3) to afford an inseparable mixture
of the silyl-protected (13Z) isomer of Compound B along with a
small amount (<10%) of the minor (13 E) isomer, which was
immediately deprotected in the next step.
The silyl-protected isomeric mixture of Compound B (2.3 mg, 3.1
mmol) was treated with 0.3 mL of a buffered solution of HF-pyridine
in THF (2:1:0.5 THF/pyridine/HF-pyridine solution from Aldrich
Chemical Co.) at 25.degree. C. After 1 h, the reaction mixture was
neutralized with saturated aqueous NaHCO.sub.3 (0.5 mL) and
extracted with EtOAc (4.times.1 mL). The combined organic extracts
were washed with saturated aqueous NaHCO.sub.3 (1 mL), dried
(Na.sub.2SO.sub.4) and the volatiles were removed in vacuo. The
residue was purified by preparative TLC (SiO.sub.2,
20.times.10.times.0.025 cm, eluting with 5% MeOH--CHCl.sub.3) to
afford title compound (13Z-isomer) along with an inseparable amount
(<10%) of the minor (13E) isomer (0.96 mg, 20% for the two
steps) as a thin film. MS (ESI.sup.+): 507.3 (M+H).sup.+; MS
(ESI.sup.-): 505.6 (M-H).sup.-.
* * * * *