U.S. patent application number 11/708849 was filed with the patent office on 2007-08-23 for processes for the convergent synthesis of calicheamicin derivatives.
This patent application is currently assigned to Wyeth. Invention is credited to Jianxin Gu, Justin Keith Moran.
Application Number | 20070197455 11/708849 |
Document ID | / |
Family ID | 38437933 |
Filed Date | 2007-08-23 |
United States Patent
Application |
20070197455 |
Kind Code |
A1 |
Moran; Justin Keith ; et
al. |
August 23, 2007 |
Processes for the convergent synthesis of calicheamicin
derivatives
Abstract
This invention describes processes for the convergent synthesis
of calicheamicin derivatives, and similar analogs using
bifunctional and trifunctional linker intermediates.
Inventors: |
Moran; Justin Keith; (Valley
Cottage, NY) ; Gu; Jianxin; (River Edge, NJ) |
Correspondence
Address: |
WYETH;PATENT LAW GROUP
5 GIRALDA FARMS
MADISON
NJ
07940
US
|
Assignee: |
Wyeth
Madison
NJ
|
Family ID: |
38437933 |
Appl. No.: |
11/708849 |
Filed: |
February 20, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60775370 |
Feb 21, 2006 |
|
|
|
Current U.S.
Class: |
514/25 ; 548/531;
560/1; 564/310 |
Current CPC
Class: |
C07D 207/46 20130101;
C07D 207/404 20130101; C07C 323/60 20130101; C07H 15/203 20130101;
A61P 35/00 20180101; C07C 319/20 20130101; C07D 309/14 20130101;
C07D 405/14 20130101; C07H 15/26 20130101; C07C 319/20 20130101;
C07C 323/60 20130101 |
Class at
Publication: |
514/25 ; 548/531;
560/1; 564/310 |
International
Class: |
C07C 69/74 20060101
C07C069/74 |
Claims
1. A process to prepare compounds of Formula (I): ##STR00048##
wherein: Z is selected from the group consisting of ##STR00049##
Alk.sup.1 is a branched or unbranched alkylene chain of 2 to 6
carbon atoms; Sp.sup.1 is selected from --S--, --O--, --CONH--,
--NHCO--, and --NR'--; Z.sup.1 is H, or alkyl of 1 to 5 carbon
atoms; Ar is 1,2-, 1,3-, or 1,4-phenylene optionally substituted
with one, two, or three groups independently selected from alkyl of
1 to 6 carbon atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1
to 4 carbon atoms, halogen, nitro, --COOR', --CONHR',
--O(CH.sub.2).sub.nCOOR', --S(CH.sub.2).sub.nCOOR',
--O(CH.sub.2).sub.nCONHR', and --S(CH.sub.2).sub.nCONHR' or a 1,2-,
1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 2,3-, 2,6-, or
2,7-naphthylidene optionally substituted with one, two, three, or
four groups independently selected from alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1 to 4 carbon
atoms, halogen, nitro, --COOR', --CONHR', --O(CH.sub.2).sub.nCOOR',
--S(CH.sub.2).sub.nCOOR', --O(CH.sub.2).sub.nCONHR', and
--S(CH.sub.2).sub.nCONHR'; n is an integer from 0 to 5; R' is a
straight or branched alkyl of 1 to 5 carbon atoms optionally
substituted by one or two groups of --OH, alkoxy of 1 to 4 carbon
atoms, thioalkoxy of 1 to 4 carbon atoms; Sp is a straight or
branched-chain divalent or trivalent alkyl radical of 1 to 18
carbon atoms, divalent or trivalent aryl or heteroaryl radical,
divalent or trivalent cycloalkyl of 3 to 18 carbon atoms or
heterocycloalkyl radical, divalent or trivalent aryl- or
heteroaryl-alkyl (C.sub.1-C.sub.18) radical, divalent or trivalent
cycloalkyl- or heterocyclo-alkyl-alkyl (C.sub.1-C.sub.18) radical
or divalent or trivalent unsaturated alkyl radical of 2 to 18
carbon atoms, wherein heteroaryl is furyl, thienyl,
N-methylpyrrolyl, pyridinyl, N-methylimidazolyl, oxazolyl,
pyrimidinyl, quinolyl, isoquinolyl, N-methylcarbazoyl,
aminocoumarinyl, or phenazinyl and wherein if Sp is a trivalent
radical, it can be additionally substituted by dialkylamino of 1 to
5 carbon atoms, alkoxy of 1 to 5 carbon atoms, hydroxy, or
alkylthio of 1 to 5 carbon atoms groups; W' is ##STR00050## R.sub.5
is --CH.sub.3, --C.sub.2H.sub.5, or --CH(CH.sub.3).sub.2; X is an
iodine or bromine atom; R.sub.5' is a hydrogen or the group RCO,
wherein R is hydrogen, branched or unbranched alkyl of 1 to 10
carbon atoms, alkylene of 2 to 10 carbon atoms, aryl of 6 to 11
carbon atoms, a (C.sub.6-C.sub.11) aryl-alkyl (C.sub.1-C.sub.5)
group, or a heteroaryl or heteroaryl-alkyl (C.sub.1-C.sub.5) group
wherein heteroaryl is defined as 2- or 3-furyl, 2- or 3-thienyl, 2-
or 3-(N-methylpyrrolyl), 2-, 3-, or 4-pyridinyl, 2-, 4-, or
5-(N-methylimidazolyl), 2-, 4-, or 5-oxazolyl, 2-, 3-, 5-, or
6-pyrimidinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, or 1-, 3-,
4-, 5-, 6-, 7-, or 8-isoquinolyl, all aryl and heteroaryl groups
optionally substituted by one or more hydroxy, amino, carboxy,
halo, nitro, (C.sub.1-C.sub.3) alkoxy of 1 to 3 carbon atoms, or
thioalkoxy of 1 to 5 carbon atoms; and Q is selected from the group
consisting of --NNHCO--, --NNHCS--, --NNHCONH--, --NNHCSNH--, and
--NO--; comprising the steps of: a. reacting a carboxylic acid of
the formula ##STR00051## with a mercapto compound of the formula
H.sub.2-Q-Sp-SH in an alcohol solvent in the presence of an alkyl
carboxylic acid, alk.sup.2CO.sub.2H where alk.sup.2 is 1 to 4
carbon atoms at about 20.degree. to 70.degree. C. for about 1 to 24
hours, wherein Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, and Sp are as
defined above, to produce a bilinker-carboxylic acid of the
formula, wherein the mercapto compound and carboxylic acid are
present in a ratio of about 1.2:1 ##STR00052## b. isolating the
bilinker-carboxylic acid of step (a); c. reacting the isolated
bilinker-carboxylic acid of step (b) with an at least a 3 fold
molar excess of N-hydroxysuccinimide, 2,3,5,6-tetrafluorophenol,
pentafluorophenol, 4-nitrophenol, 2,4-dinitrophenol, or
N-hydroxysulfosuccinimide in the presence of
1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or
N,N'-disuccinimidyl carbonate in an inert solvent containing 0-50%
N,N-dimethylformamide (DMF) to generate a trilinker-activated ester
of the formula ##STR00053## where Z is hereinbefore defined; d.
reacting the trilinker-activated ester formed in step c with a
methyltrithio antitumor antibiotic CH.sub.3--S--S--S--W' in the
presence of a base or an organic base with a methyltrithio
antitumor antibiotic CH.sub.3--S--S--S--W' in an inert organic
solvent to generate an activated ester of the formula below,
wherein the trilinker-activated ester in step c and the
CH.sub.3--S--S--S--W' are in a ratio of 3.3:1 and the temperature
of the reaction is .ltoreq.5.degree. C. ##STR00054## e. isolating
the activated ester of step (d) and purifying to yield antitumor
antibiotics of Formula (I) ##STR00055##
2. The process of claim 1 wherein the purifying of step (e)
comprises the use of reverse phase high performance liquid
chromatography having a mobile phase of about pH 7.0 to 9.0
followed by normal phase chromatography.
3. A process according to claim 1, wherein alk.sup.1 is 3 carbon
atoms; Sp.sup.1 is --O--; Z.sup.1 is methyl; Ar is unsubstituted
1,4-phenylene; Sp is 4 carbon atoms; R.sub.3 is H; Q is NNHC(O)--;
##STR00056## R.sub.5 is C.sub.2H.sub.5; R.sub.5' is --C(O)--R; R is
methyl; and Z is ##STR00057##
4. A process to prepare antitumor antibiotics of Formula (I):
##STR00058## wherein: Z is selected from the group consisting of
##STR00059## Alk.sup.1 is a branched or unbranched alkylene chain
of 2 to 6 carbon atoms; Sp.sup.1 is selected from --S--, --O--,
--CONH--, --NHCO--, and --NR'--; Z.sup.1 is H, or alkyl of 1 to 5
carbon atoms; Ar is 1,2-, 1,3-, or 1,4-phenylene optionally
substituted with one, two, or three groups independently selected
from alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 5 carbon atoms,
thioalkoxy of 1 to 4 carbon atoms, halogen, nitro, --COOR',
--CONHR', --O(CH.sub.2).sub.nCOOR', --S(CH.sub.2).sub.nCOOR',
--O(CH.sub.2).sub.nCONHR', and --S(CH.sub.2).sub.nCONHR' or a 1,2-,
1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 2,3-, 2,6-, or
2,7-naphthylidene optionally substituted with one, two, three, or
four groups independently selected from alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1 to 4 carbon
atoms, halogen, nitro, --COOR', --CONHR', --O(CH.sub.2).sub.nCOOR',
--S(CH.sub.2).sub.nCOOR', --O(CH.sub.2).sub.nCONHR', and
--S(CH.sub.2).sub.nCONHR'; n is an integer from 0 to 5; R' is a
straight or branched alkyl of 1 to 5 carbon atoms optionally
substituted by one or two groups of --OH, alkoxy of 1 to 4 carbon
atoms, thioalkoxy of 1 to 4 carbon atoms; Sp is a straight or
branched-chain divalent or trivalent alkyl radical of 1 to 18
carbon atoms, divalent or trivalent aryl or heteroaryl radical,
divalent or trivalent cycloalkyl of 3 to 18 carbon atoms or
heterocycloalkyl radical, divalent or trivalent aryl- or
heteroaryl-alkyl (C.sub.1-C.sub.18) radical, divalent or trivalent
cycloalkyl- or heterocyclo-alkyl-alkyl (C.sub.1-C.sub.18) radical
or divalent or trivalent unsaturated alkyl radical of 2 to 18
carbon atoms, wherein heteroaryl is furyl, thienyl,
N-methylpyrrolyl, pyridinyl, N-methylimidazolyl, oxazolyl,
pyrimidinyl, quinolyl, isoquinolyl, N-methylcarbazoyl,
aminocoumarinyl, or phenazinyl and wherein if Sp is a trivalent
radical, it can be additionally substituted by dialkylamino of 1 to
5 carbon atoms, alkoxy of 1 to 5 carbon atoms, hydroxy, or
alkylthio of 1 to 5 carbon atoms groups; W' is ##STR00060## R.sub.5
is --CH.sub.3, --C.sub.2H.sub.5, or --CH(CH.sub.3).sub.2; X is an
iodine or bromine atom; R.sub.5' is a hydrogen or the group RCO,
wherein R is hydrogen, branched or unbranched alkyl of 1 to 10
carbon atoms, alkylene of 2 to 10 carbon atoms, aryl of 6 to 11
carbon atoms, a (C.sub.6-C.sub.11) aryl-alkyl (C.sub.1-C.sub.5)
group, or a heteroaryl or heteroaryl-alkyl (C.sub.1-C.sub.5) group
wherein heteroaryl is defined as 2- or 3-furyl, 2- or 3-thienyl, 2-
or 3-(N-methylpyrrolyl), 2-, 3-, or 4-pyridinyl, 2-, 4-, or
5-(N-methylimidazolyl), 2-, 4-, or 5-oxazolyl, 2-, 3-, 5-, or
6-pyrimidinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, or 1-, 3-,
4-, 5-, 6-, 7-, or 8-isoquinolyl, all aryl and heteroaryl groups
optionally substituted by one or more hydroxy, amino, carboxy,
halo, nitro, (C.sub.1-C.sub.3) alkoxy of 1 to 3 carbon atoms, or
thioalkoxy of 1 to 5 carbon atoms; and Q is selected from the group
consisting of --NNHCO--, --NNHCS--, --NNHCONH--, --NNHCSNH--, and
--NO--; comprising the steps of: a. reacting a carboxylic acid of
the formula ##STR00061## with a mercapto compound of the formula
H.sub.2Q-Sp-SH in an alcohol solvent in the presence of an alkyl
carboxylic acid, alk.sup.2CO.sub.2H where alk.sup.2 is 1 to 4
carbon atoms at about 20.degree. to 70.degree. C. for about 1 to 24
hours, wherein Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, and Sp are as
defined above, to produce a bilinker-carboxylic acid of the
formula, wherein the mercapto compound and carboxylic acid are
present in a ratio of about 1.2:1 ##STR00062## b. isolating the
bilinker-carboxylic acid of step (a); c. reacting the isolated
bilinker-carboxylic acid of step (b) with a methyltrithio antitumor
antibiotic CH.sub.3--S--S--S--W' in the presence of a base or an
organic base with a methyltrithio antitumor antibiotic
CH.sub.3--S--S--S--W' in an inert organic solvent; d. reacting the
compound of step (c) with an at least a 3 fold molar excess of
N-hydroxysuccinimide, 2,3,5,6-tetrafluorophenol, pentafluorophenol,
4-nitrophenol, 2,4-dinitrophenol, or N-hydroxysulfosuccinimide in
the presence of 1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or
N,N'-disuccinimidyl carbonate in an inert solvent containing 0-50%
N,N-dimethylformamide (DMF) and purifying to yield antitumor
antibiotics of Formula (I) ##STR00063##
5. The process of claim 4 wherein the purifying of step (e)
comprises the use of a reverse phase high performance liquid
chromatography having a mobile phase of about pH 7.0 to 9.0
followed with a normal phase chromatography.
6. A process according to claim 4, wherein alk.sup.1 is 3 carbon
atoms; Sp.sup.1 is --O--; Z.sup.1 is methyl; Ar is unsubstituted
1,4-phenylene; Sp is 4 carbon atoms; R.sub.3 is H; Q is NNHC(O)--;
##STR00064## R.sub.5 is C.sub.2H.sub.5; R.sub.5' is --C(O)--R; R is
methyl; and Z is ##STR00065##
7. A process to prepare antitumor antibiotics of Formula (I):
##STR00066## wherein: Z is selected from the group consisting of
##STR00067## Alk.sup.1 is a branched or unbranched alkylene chain
of 2 to 6 carbon atoms; Sp.sup.1 is selected from --S--, --O--,
--CONH--, --NHCO--, and --NR'--; Z.sup.1 is H, or alkyl of 1 to 5
carbon atoms; Ar is 1,2-, 1,3-, or 1,4-phenylene optionally
substituted with one, two, or three groups independently selected
from alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 5 carbon atoms,
thioalkoxy of 1 to 4 carbon atoms, halogen, nitro, --COOR',
--CONHR', --O(CH.sub.2).sub.nCOOR', --S(CH.sub.2).sub.nCOOR',
--O(CH.sub.2).sub.nCONHR', and --S(CH.sub.2).sub.nCONHR' or a 1,2-,
1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 2,3-, 2,6-, or
2,7-naphthylidene optionally substituted with one, two, three, or
four groups independently selected from alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1 to 4 carbon
atoms, halogen, nitro, --COOR', --CONHR', --O(CH.sub.2).sub.nCOOR',
--S(CH.sub.2).sub.nCOOR', --O(CH.sub.2).sub.nCONHR', and
--S(CH.sub.2).sub.nCONHR'; n is an integer from 0 to 5; R' is a
straight or branched alkyl of 1 to 5 carbon atoms optionally
substituted by one or two groups of --OH, alkoxy of 1 to 4 carbon
atoms, thioalkoxy of 1 to 4 carbon atoms; Sp is a straight or
branched-chain divalent or trivalent alkyl radical of 1 to 18
carbon atoms, divalent or trivalent aryl or heteroaryl radical,
divalent or trivalent cycloalkyl of 3 to 18 carbon atoms or
heterocycloalkyl radical, divalent or trivalent aryl- or
heteroaryl-alkyl (C.sub.1-C.sub.18) radical, divalent or trivalent
cycloalkyl- or heterocyclo-alkyl-alkyl (C.sub.1-C.sub.18) radical
or divalent or trivalent unsaturated alkyl radical of 2 to 18
carbon atoms, wherein heteroaryl is furyl, thienyl,
N-methylpyrrolyl, pyridinyl, N-methylimidazolyl, oxazolyl,
pyrimidinyl, quinolyl, isoquinolyl, N-methylcarbazoyl,
aminocoumarinyl, or phenazinyl and wherein if Sp is a trivalent
radical, it can be additionally substituted by dialkylamino of 1 to
5 carbon atoms, alkoxy of 1 to 5 carbon atoms, hydroxy, or
alkylthio of 1 to 5 carbon atoms groups; W' is ##STR00068## R.sub.5
is --CH.sub.3, --C.sub.2H.sub.5, or --CH(CH.sub.3).sub.2; X is an
iodine or bromine atom; R.sub.5' is a hydrogen or the group RCO,
wherein R is hydrogen, branched or unbranched alkyl of 1 to 10
carbon atoms, alkylene of 2 to 10 carbon atoms, aryl of 6 to 11
carbon atoms, a (C.sub.6-C.sub.11) aryl-alkyl (C.sub.1-C.sub.5)
group, or a heteroaryl or heteroaryl-alkyl (C.sub.1-C.sub.5) group
wherein heteroaryl is defined as 2- or 3-furyl, 2- or 3-thienyl, 2-
or 3-(N-methylpyrrolyl), 2-, 3-, or 4-pyridinyl, 2-, 4-, or
5-(N-methylimidazolyl), 2-, 4-, or 5-oxazolyl, 2-, 3-, 5-, or
6-pyrimidinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, or 1-, 3-,
4-, 5-, 6-, 7-, or 8-isoquinolyl, all aryl and heteroaryl groups
optionally substituted by one or more hydroxy, amino, carboxy,
halo, nitro, (C.sub.1-C.sub.3) alkoxy of 1 to 3 carbon atoms, or
thioalkoxy of 1 to 5 carbon atoms; and Q is selected from the group
consisting of --NNHCO--, --NNHCS--, --NNHCONH--, --NNHCSNH--, and
--NO--; comprising the steps of: a. reacting a carboxylic acid of
the formula ##STR00069## with a mercapto compound of the formula
H.sub.2-Q-Sp-SH in an alcohol solvent in the presence of an alkyl
carboxylic acid, alk.sup.2CO.sub.2H where alk.sup.2 is 1 to 4
carbon atoms at about 20.degree. to 70.degree. C. for about 1 to 24
hours, wherein Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, and Sp are as
defined above, to produce a bilinker-carboxylic acid of the
formula, ##STR00070## b. isolating the bilinker-carboxylic acid of
step (a); c. reacting the isolated bilinker-carboxylic acid of step
(b) with an at least of N-hydroxysuccinimide,
2,3,5,6-tetrafluorophenol, pentafluorophenol, 4-nitrophenol,
2,4-dinitrophenol, or N-hydroxysulfosuccinimide in the presence of
1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or
N,N'-disuccinimidyl carbonate in an inert solvent containing 0-50%
N,N-dimethylformamide (DMF) to generate a trilinker-activated ester
of the formula ##STR00071## where Z is hereinbefore defined d.
reacting the trilinker-activated ester in step c in the presence of
a base or an organic base with a methyltrithio antitumor antibiotic
CH.sub.3--S--S--S--W' in an inert organic solvent to generate an
activated ester of the formula ##STR00072## e. isolating the
activated ester of step (d) and purifying to yield antitumor
antibiotics of Formula (I) ##STR00073##
8. A process according to claim 1, wherein alk.sup.1 is 3 carbon
atoms; Sp.sup.1 is --O--; Z.sup.1 is methyl; Ar is unsubstituted
1,4-phenylene; Sp is 4 carbon atoms; R.sub.3 is H; Q is NNHC(O)--;
##STR00074## R.sub.5 is C.sub.2H.sub.5; R.sub.5' is --C(O)--R; R is
methyl; and Z is ##STR00075##
9. A process according to claim 1, wherein Alk.sup.1 is an alkylene
of 2 to 5 carbon atoms, and Sp.sup.1 is an oxygen atom.
10. A process according to claim 9, wherein Alk.sup.1 is an
alkylene of 3 carbon atoms.
11. A process according to claim 1, wherein Z.sup.1 is alkyl of 1
to 3 carbon atoms.
12. A process according to claim 1, wherein Ar is 1,2-, 1,3-, or
1,4-phenylene, or 1,2-, 1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 2,3-,
2,6-, or 2,7-naphthylidene.
13. A process according to claim 12, wherein Ar is
1,4-phenylene.
14. A process according to claim 1, wherein Q is --NNHCO--.
15. A process according to claim 1, wherein Sp is straight or
branched-chain divalent or trivalent alkyl radical of 1 to 12
carbon atoms.
16. A process according to claim 15, wherein Sp is straight or
branched-chain divalent or trivalent alkyl radical of 1 to 6 carbon
atoms.
17. The process according to claim 1, wherein the alcohol solvent
is methanol.
18. The process according to claim 1, wherein the inert solvent is
acetonitrile.
19. The process according to claim 1, wherein alkyl carboxylic acid
is acetic acid.
20. The process according to claim 1, wherein the inert organic
solvent is acetonitrile.
21. A process according to claim 1, wherein Z is ##STR00076##
22. A process to prepare trilinker-activated esters of the formula:
##STR00077## wherein: Z is selected from the group consisting of
##STR00078## Alk.sup.1 is a branched or unbranched alkylene chain
of 2 to 6 carbon atoms; Sp.sup.1 is selected from --S--, --O--,
--CONH--, --NHCO--, and --NR'--; Z.sup.1 is H, or alkyl of 1 to 5
carbon atoms; Ar is 1,2-, 1,3-, or 1,4-phenylene optionally
substituted with one, two, or three groups independently selected
from alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 5 carbon atoms,
thioalkoxy of 1 to 4 carbon atoms, halogen, nitro, --COOR',
--CONHR', --O(CH.sub.2).sub.nCOOR', --S(CH.sub.2).sub.nCOOR',
--O(CH.sub.2).sub.nCONHR', and --S(CH.sub.2).sub.nCONHR' or a 1,2-,
1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 2,3-, 2,6-, or
2,7-naphthylidene optionally substituted with one, two, three, or
four groups independently selected from alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1 to 4 carbon
atoms, halogen, nitro, --COOR', --CONHR', --O(CH.sub.2).sub.nCOOR',
--S(CH.sub.2).sub.nCOOR', --O(CH.sub.2).sub.nCONHR', and
--S(CH.sub.2).sub.nCONHR'; n is an integer from 0 to 5; R' is a
straight or branched alkyl of 1 to 5 carbon atoms optionally
substituted by one or two groups of --OH, alkoxy of 1 to 4 carbon
atoms, thioalkoxy of 1 to 4 carbon atoms; Sp is a straight or
branched-chain divalent or trivalent alkyl radical of 1 to 18
carbon atoms, divalent or trivalent aryl or heteroaryl radical,
divalent or trivalent cycloalkyl of 3 to 18 carbon atoms or
heterocycloalkyl radical, divalent or trivalent aryl- or
heteroaryl-alkyl (C.sub.1-C.sub.18) radical, divalent or trivalent
cycloalkyl- or heterocyclo-alkyl-alkyl (C.sub.1-C.sub.18) radical
or divalent or trivalent unsaturated alkyl radical of 2 to 18
carbon atoms, wherein heteroaryl is furyl, thienyl,
N-methylpyrrolyl, pyridinyl, N-methylimidazolyl, oxazolyl,
pyrimidinyl, quinolyl, isoquinolyl, N-methylcarbazoyl,
aminocoumarinyl, or phenazinyl and wherein if Sp is a trivalent
radical, it can be additionally substituted by dialkylamino of 1 to
5 carbon atoms, alkoxy of 1 to 5 carbon atoms, hydroxy, or
alkylthio of 1 to 5 carbon atoms groups; W' is ##STR00079## R.sub.5
is --CH.sub.3, --C.sub.2H.sub.5, or --CH(CH.sub.3).sub.2; X is an
iodine or bromine atom; R.sub.5' is a hydrogen or the group RCO,
wherein R is hydrogen, branched or unbranched alkyl of 1 to 10
carbon atoms, alkylene of 2 to 10 carbon atoms, aryl of 6 to 11
carbon atoms, a (C.sub.6-C.sub.11) aryl-alkyl (C.sub.1-C.sub.5)
group, or a heteroaryl or heteroaryl-alkyl (C.sub.1-C.sub.5) group
wherein heteroaryl is defined as 2- or 3-furyl, 2- or 3-thienyl, 2-
or 3-(N-methylpyrrolyl), 2-, 3-, or 4-pyridinyl, 2-, 4-, or
5-(N-methylimidazolyl), 2-, 4-, or 5-oxazolyl, 2-, 3-, 5-, or
6-pyrimidinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, or 1-, 3-,
4-, 5-, 6-, 7-, or 8-isoquinolyl, all aryl and heteroaryl groups
optionally substituted by one or more hydroxy, amino, carboxy,
halo, nitro, (C.sub.1-C.sub.3) alkoxy of 1 to 3 carbon atoms, or
thioalkoxy of 1 to 5 carbon atoms; and Q is selected from the group
consisting of --NNHCO--, --NNHCS--, --NHNCONH--, --NNCSNH--, and
--NO--; comprising the steps of: a. reacting a carboxylic acid of
the formula ##STR00080## with a mercapto compound of the formula
H.sub.2Q-Sp-SH in an alcohol solvent in the presence of an alkyl
carboxylic acid, alk.sup.2CO.sub.2H where alk.sup.2 is 1 to 4
carbon atoms at about 20.degree. to 70.degree. C. for about 1 to 24
hours, wherein Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, and Sp are as
defined above, to produce a bilinker-carboxylic acid of the formula
##STR00081## b. isolating the bilinker-carboxylic acid of step (a);
c. reacting the bilinker-carboxylic acid from step (b) in the
presence of a base or an organic base with a methyltrithio
antitumor antibiotic CH.sub.3--S--S--S--W' in an inert organic
solvent to generate a bilinker-methyltrithio antitumor antibiotic
of the formula ##STR00082## d. reacting the isolated
bilinker-methyltrithio antitumor antibiotic of step (c) with
N-hydroxysuccinimide, 2,3,5,6-tetrafluorophenol, pentafluorophenol,
4-nitrophenol, 2,4-dinitrophenol, or N-hydroxysulfosuccinimide in
the presence of 1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or
N,N'-disuccinimidyl carbonate in an inert solvent containing 0-50%
N,N-dimethylformamide (DMF) to generate a trilinker-activated ester
of the formula ##STR00083##
23. The process according to claim 17, wherein Alk.sup.1 is an
alkylene of 2 to 5 carbon atoms, and Sp.sup.1 is an oxygen
atom.
24. A process for the preparation of trifunctional linker
intermediates, of the formula ##STR00084## wherein: Alk.sup.1 is a
branched or unbranched alkylene chain of 2 to 6 carbon atoms;
Sp.sup.1 is selected from --S--, --O--, --CONH--, --NHCO--, and
--NR'--; Z.sup.1 is H, or alkyl of 1 to 5 carbon atoms; Ar is 1,2-,
1,3-, or 1,4-phenylene optionally substituted with one, two, or
three groups independently selected from alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1 to 4 carbon
atoms, halogen, nitro, --COOR', --CONHR', --O(CH.sub.2).sub.nCOOR',
--S(CH.sub.2).sub.nCOOR', --O(CH.sub.2).sub.nCONHR', and
--S(CH.sub.2).sub.nCONHR' or a 1,2-, 1,3-, 1,4-, 1,5-, 1,6-, 1,7-,
1,8-, 2,3-, 2,6-, or 2,7-naphthylidene optionally substituted with
one, two, three, or four groups independently selected from alkyl
of 1 to 6 carbon atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy
of 1 to 4 carbon atoms, halogen, nitro, --COOR', --CONHR',
--O(CH.sub.2).sub.nCOOR', --S(CH.sub.2).sub.nCOOR',
--O(CH.sub.2).sub.nCONHR', and --S(CH.sub.2).sub.nCONHR'; n is an
integer from 0 to 5; R' is a straight or branched alkyl of 1 to 5
carbon atoms optionally substituted by one or two groups of --OH,
alkoxy of 1 to 4 carbon atoms, thioalkoxy of 1 to 4 carbon atoms;
Sp is a straight or branched-chain divalent or trivalent alkyl
radical of 1 to 18 carbon atoms, divalent or trivalent aryl or
heteroaryl radical, divalent or trivalent cycloalkyl of 3 to 18
carbon atoms or heterocycloalkyl radical, divalent or trivalent
aryl- or heteroaryl-alkyl (C.sub.1-C.sub.18) radical, divalent or
trivalent cycloalkyl- or heterocyclo-alkyl-alkyl (C.sub.1-C.sub.18)
radical or divalent or trivalent unsaturated alkyl radical of 2 to
18 carbon atoms, wherein heteroaryl is furyl, thienyl,
N-methylpyrrolyl, pyridinyl, N-methylimidazolyl, oxazolyl,
pyrimidinyl, quinolyl, isoquinolyl, N-methylcarbazoyl,
aminocoumarinyl, or phenazinyl and wherein if Sp is a trivalent
radical, it can be additionally substituted by dialkylamino of 1 to
5 carbon atoms, alkoxy of 1 to 5 carbon atoms, hydroxy, or
alkylthio of 1 to 5 carbon atoms groups; Q is selected from the
group consisting of --NNHCO--, --NNHCS--, and --NNHCONH--; Z is
selected from the group consisting of ##STR00085## comprising the
steps of: a. reacting a carboxylic acid of the formula ##STR00086##
with a mercapto compound of the formula H.sub.2Q-Sp-SH in an
alcohol solvent in the presence of an alkyl carboxylic acid,
alk.sup.2CO.sub.2H, where alk.sup.2 is 1 to 4 carbon atoms at about
20.degree. to 70.degree. C. for about 1 to 24 hours, wherein
Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, and Sp are as defined above,
to produce a bilinker-carboxylic acid of the formula ##STR00087##
b. isolating the bilinker-carboxylic acid of step (a); c. reacting
the isolated bilinker-carboxylic acid of step (b) with
N-hydroxysuccinimide, 2,3,5,6-tetrafluorophenol, pentafluorophenol,
4-nitrophenol, 2,4-dinitrophenol, or N-hydroxysulfosuccinimide in
the presence of 1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or
N,N'-disuccinimidyl carbonate in an inert solvent containing 0-50%
DMF to generate trifunctional linker intermediates, of the formula
##STR00088##
Description
[0001] This application claims priority from U.S. Provisional
Application 60/775,370 filed Feb. 21, 2006 entitled "Processes For
The Convergent Synthesis of Calicheamicin Derivatives" the content
of which is incorporated herein in its entirety to the extent that
it is consistent with this invention and application.
FIELD OF THE INVENTION
[0002] This invention describes processes for the convergent
synthesis of calicheamicin derivatives, and similar analogs using
bifunctional and trifunctional linker intermediates.
BACKGROUND OF THE INVENTION
[0003] The potent family of antibacterial and antitumor agents
known collectively as the calicheamicins or the LL-E33288 complex,
are disclosed in U.S. Pat. No. 4,970,198 (1990). The compounds
contain a methyltrisulfide that can be reacted with appropriate
thiols to form disulfides while at the same time introducing a
functional group such as a hydrazide or similar nucleophile.
Examples of this reaction with the calicheamicins are given in U.S.
Pat. No. 5,053,394. U.S. Pat. No. 5,770,701 is directed to a
process for preparing targeted forms of disulfide compounds of the
LL-E33288 complex. A linker, 4-(4-acetyl-phenoxy)butanoic acid, is
condensed with calicheamicin N-acetyl gamma dimethyl hydrazide to
afford the carboxylic acid-hydrazone which is further treated with
N-hydroxysuccinimide to give the OSu ester (N-succinimidyloxy)
which is ready for conjugation with a chosen biomacromolecule.
[0004] Previously disclosed synthetic methods for constructing
calicheamicin derivatives are complicated by multiple calicheamicin
containing synthetic steps having low overall yields. The
calicheamicin moiety is inherently toxic, and when already part of
a synthetic target necessitates increased safety precautions which
must be observed during manipulation and purification of the
products of each of the synthetic steps. The claimed process
provides a method of fewer steps involving the calicheamicin moiety
with increased yields.
SUMMARY OF THE INVENTION
[0005] Discussed herein is a process to prepare calicheamicin
derivatives of Formula (I):
##STR00001##
wherein:
Z is selected from the group consisting of
##STR00002##
[0006] Alk.sup.1 is a branched or unbranched alkylene chain of 2 to
6 carbon atoms;
Sp.sup.1 is selected from --S--, --O--, --CONH--, --NHCO--, and
--NR'--;
Z.sup.1 is H, or alkyl of 1 to 5 carbon atoms;
[0007] Ar is 1,2-, 1,3-, or 1,4-phenylene optionally substituted
with one, two, or three groups independently selected from alkyl of
1 to 6 carbon atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1
to 4 carbon atoms, halogen, nitro, --COOR', --CONHR',
--O(CH.sub.2).sub.nCOOR', --S(CH.sub.2).sub.nCOOR',
--O(CH.sub.2).sub.nCONHR', and --S(CH.sub.2).sub.nCONHR' or a 1,2-,
1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 2,3-, 2,6-, or
2,7-naphthylidene optionally substituted with one, two, three, or
four groups independently selected from alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1 to 4 carbon
atoms, halogen, nitro, --COOR', --CONHR', --O(CH.sub.2).sub.nCOOR',
--S(CH.sub.2).sub.nCOOR', --O(CH.sub.2).sub.nCONHR', and
--S(CH.sub.2).sub.nCONHR'; n is an integer from 0 to 5;
R' is a straight or branched alkyl of 1 to 5 carbon atoms
optionally substituted by one or two groups of --OH, alkoxy of 1 to
4 carbon atoms, thioalkoxy of 1 to 4 carbon atoms;
[0008] Sp is a straight or branched-chain divalent or trivalent
alkyl radical of 1 to 18 carbon atoms, divalent or trivalent aryl
or heteroaryl radical, divalent or trivalent cycloalkyl of 3 to 18
carbon atoms or heterocycloalkyl radical, divalent or trivalent
aryl- or heteroaryl-alkyl (C.sub.1-C.sub.18) radical, divalent or
trivalent cycloalkyl- or heterocyclo-alkyl-alkyl (C.sub.1-C.sub.18)
radical or divalent or trivalent unsaturated alkyl radical of 2 to
18 carbon atoms, wherein heteroaryl is furyl, thienyl,
N-methylpyrrolyl, pyridinyl, N-methylimidazolyl, oxazolyl,
pyrimidinyl, quinolyl, isoquinolyl, N-methylcarbazoyl,
aminocoumarinyl, or phenazinyl and wherein if Sp is a trivalent
radical, it can be additionally substituted by dialkylamino of 1 to
5 carbon atoms, alkoxy of 1 to 5 carbon atoms, hydroxy, or
alkylthio of 1 to 5 carbon atoms groups;
W' is
##STR00003##
[0009] R.sub.5 is --CH.sub.3, --C.sub.2H.sub.5, or
--CH(CH.sub.3).sub.2;
X is an iodine or bromine atom;
[0010] R.sub.5' is a hydrogen or the group RCO, wherein R is
hydrogen, branched or unbranched alkyl of 1 to 10 carbon atoms,
alkylene of 2 to 10 carbon atoms, aryl of 6 to 11 carbon atoms, a
(C.sub.6-C.sub.11) aryl-alkyl (C.sub.1-C.sub.5) group, or a
heteroaryl or heteroaryl-alkyl (C.sub.1-C.sub.5) group wherein
heteroaryl is defined as 2- or 3-furyl, 2- or 3-thienyl, 2- or
3-(N-methylpyrrolyl), 2-, 3-, or 4-pyridinyl, 2-, 4-, or
5-(N-methylimidazolyl), 2-, 4-, or 5-oxazolyl, 2-, 3-, 5-, or
6-pyrimidinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, or 1-, 3-,
4-, 5-, 6-, 7-, or 8-isoquinolyl, all aryl and heteroaryl groups
optionally substituted by one or more hydroxy, amino, carboxy,
halo, nitro, (C.sub.1-C.sub.3)alkoxy of 1 to 3 carbon atoms, or
thioalkoxy of 1 to 5 carbon atoms; and
Q is selected from the group consisting of --NNHCO--, --NNHCS--,
--NNHCONH--, --NNHCSNH--, and --NO--;
[0011] comprising the steps of: a. reacting a carboxylic acid of
the formula
##STR00004##
with a mercapto compound of the formula
H.sub.2Q-Sp-SH
in an alcohol solvent in the presence of an alkyl carboxylic acid,
alk.sup.2CO.sub.2H where alk.sup.2 is 1 to 4 carbon atoms at about
20.degree. to 70.degree. C. for about 1 to 24 hours, wherein
Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, and Sp are as defined above,
to produce a bilinker-carboxylic acid of the formula, wherein the
mercapto compound and carboxylic acid are present in a ratio of
about 1.2:1
##STR00005##
b. isolating the bilinker-carboxylic acid of step (a); c. reacting
the isolated bilinker-carboxylic acid of step (b) with an at least
a 3 fold molar excess of N-hydroxysuccinimide,
2,3,5,6-tetrafluorophenol, pentafluorophenol, 4-nitrophenol,
2,4-dinitrophenol, or N-hydroxysulfosuccinimide in the presence of
1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or
N,N'-disuccinimidyl carbonate in an inert solvent containing 0-50%
N,N-dimethylformamide (DMF) to generate a trilinker-activated ester
of the formula
##STR00006##
where Z is hereinbefore defined; d. reacting the
trilinker-activated ester formed in step with a methyltrithio
antitumor antibiotic CH.sub.3--S--S--S--W' in the presence of a
base or an organic base with a methyltrithio antitumor antibiotic
CH.sub.3--S--S--S--W' in an inert organic solvent to generate an
activated ester of the formula below, wherein the
trilinker-activated ester in step c and the CH.sub.3--S--S--S--W'
are in a ratio of 3.3:1 and the temperature of the reaction is
.ltoreq.5.degree. C.
##STR00007##
e. isolating the activated ester of step (d) and purifying to yield
antitumor antibiotics of Formula (I)
##STR00008##
[0012] An additional process discussed includes a method of
preparing antitumor antibiotics of Formula (I):
##STR00009##
wherein:
Z is selected from the group consisting of
##STR00010##
[0013] Alk.sup.1 is a branched or unbranched alkylene chain of 2 to
6 carbon atoms;
Sp.sup.1 is selected from --S--, --O--, --CONH--, --NHCO--, and
--NR'--;
Z.sup.1 is H, or alkyl of 1 to 5 carbon atoms;
[0014] Ar is 1,2-, 1,3-, or 1,4-phenylene optionally substituted
with one, two, or three groups independently selected from alkyl of
1 to 6 carbon atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1
to 4 carbon atoms, halogen, nitro, --COOR', --CONHR',
--O(CH.sub.2).sub.nCOOR', --S(CH.sub.2).sub.nCOOR',
--O(CH.sub.2).sub.nCONHR', and --S(CH.sub.2).sub.nCONHR' or a 1,2-,
1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 2,3-, 2,6-, or
2,7-naphthylidene optionally substituted with one, two, three, or
four groups independently selected from alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1 to 4 carbon
atoms, halogen, nitro, --COOR', --CONHR', --O(CH.sub.2).sub.nCOOR',
--S(CH.sub.2).sub.nCOOR', --O(CH.sub.2).sub.nCONHR', and
--S(CH.sub.2).sub.nCONHR'; n is an integer from 0 to 5;
R' is a straight or branched alkyl of 1 to 5 carbon atoms
optionally substituted by one or two groups of --OH, alkoxy of 1 to
4 carbon atoms, thioalkoxy of 1 to 4 carbon atoms;
[0015] Sp is a straight or branched-chain divalent or trivalent
alkyl radical of 1 to 18 carbon atoms, divalent or trivalent aryl
or heteroaryl radical, divalent or trivalent cycloalkyl of 3 to 18
carbon atoms or heterocycloalkyl radical, divalent or trivalent
aryl- or heteroaryl-alkyl (C.sub.1-C.sub.18) radical, divalent or
trivalent cycloalkyl- or heterocyclo-alkyl-alkyl (C.sub.1-C.sub.18)
radical or divalent or trivalent unsaturated alkyl radical of 2 to
18 carbon atoms, wherein heteroaryl is furyl, thienyl,
N-methylpyrrolyl, pyridinyl, N-methylimidazolyl, oxazolyl,
pyrimidinyl, quinolyl, isoquinolyl, N-methylcarbazoyl,
aminocoumarinyl, or phenazinyl and wherein if Sp is a trivalent
radical, it can be additionally substituted by dialkylamino of 1 to
5 carbon atoms, alkoxy of 1 to 5 carbon atoms, hydroxy, or
alkylthio of 1 to 5 carbon atoms groups;
W' is
##STR00011##
[0016] R.sub.5 is --CH.sub.3, --C.sub.2H.sub.5, or
--CH(CH.sub.3).sub.2;
X is an iodine or bromine atom;
[0017] R.sub.5' is a hydrogen or the group RCO, wherein R is
hydrogen, branched or unbranched alkyl of 1 to 10 carbon atoms,
alkylene of 2 to 10 carbon atoms, aryl of 6 to 11 carbon atoms, a
(C.sub.6-C.sub.11) aryl-alkyl (C.sub.1-C.sub.5) group, or a
heteroaryl or heteroaryl-alkyl (C.sub.1-C.sub.5) group wherein
heteroaryl is defined as 2- or 3-furyl, 2- or 3-thienyl, 2- or
3-(N-methylpyrrolyl), 2-, 3-, or 4-pyridinyl, 2-, 4-, or
5-(N-methylimidazolyl), 2-, 4-, or 5-oxazolyl, 2-, 3-, 5-, or
6-pyrimidinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, or 1-, 3-,
4-, 5-, 6-, 7-, or 8-isoquinolyl, all aryl and heteroaryl groups
optionally substituted by one or more hydroxy, amino, carboxy,
halo, nitro, (C.sub.1-C.sub.3) alkoxy of 1 to 3 carbon atoms, or
thioalkoxy of 1 to 5 carbon atoms; and
Q is selected from the group consisting of --NNHCO--, --NNHCS--,
--NNHCONH--, --NNHCSNH--, and --NO--;
[0018] comprising the steps of: a. reacting a carboxylic acid of
the formula
##STR00012##
with a mercapto compound of the formula
H.sub.2Q-Sp-SH
in an alcohol solvent in the presence of an alkyl carboxylic acid,
alk.sup.2CO.sub.2H where alk.sup.2 is 1 to 4 carbon atoms at about
20.degree. to 70.degree. C. for about 1 to 24 hours, wherein
Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, and Sp are as defined above,
to produce a bilinker-carboxylic acid of the formula, wherein the
mercapto compound and carboxylic acid are present in a ratio of
about 1.2:1
##STR00013##
b. isolating the bilinker-carboxylic acid of step (a); c. reacting
the isolated bilinker-carboxylic acid of step (b) with
CH.sub.3--S--S--S--W' in the presence of a base or an organic base
with a methyltrithio antitumor antibiotic CH.sub.3--S--S--S--W' in
an inert organic solvent; d. reacting the compound of step (c) with
an at least a 3 fold molar excess of N-hydroxysuccinimide,
2,3,5,6-tetrafluorophenol, pentafluorophenol, 4-nitrophenol,
2,4-dinitrophenol, or N-hydroxysulfosuccinimide in the presence of
1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or
N,N'-disuccinimidyl carbonate in an inert solvent containing 0-50%
N,N-dimethylformamide (DMF) and purifying to yield compounds of
Formula (I)
##STR00014##
[0019] An additional process includes preparation of compounds of
Formula (I):
##STR00015##
wherein:
Z is selected from the group consisting of
##STR00016##
[0020] Alk.sup.1 is a branched or unbranched alkylene chain of 2 to
6 carbon atoms;
Sp.sup.1 is selected from --S--, --O--, --CONH--, --NHCO--, and
--NR'--;
Z.sup.1 is H, or alkyl of 1 to 5 carbon atoms;
[0021] Ar is 1,2-, 1,3-, or 1,4-phenylene optionally substituted
with one, two, or three groups independently selected from alkyl of
1 to 6 carbon atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1
to 4 carbon atoms, halogen, nitro, --COOR', --CONHR',
--O(CH.sub.2).sub.nCOOR', --S(CH.sub.2).sub.nCOOR',
--O(CH.sub.2).sub.nCONHR', and --S(CH.sub.2).sub.nCONHR' or a 1,2-,
1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 2,3-, 2,6-, or
2,7-naphthylidene optionally substituted with one, two, three, or
four groups independently selected from alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1 to 4 carbon
atoms, halogen, nitro, --COOR', --CONHR', --O(CH.sub.2).sub.nCOOR',
--S(CH.sub.2).sub.nCOOR', --O(CH.sub.2).sub.nCONHR', and
--S(CH.sub.2).sub.nCONHR'; n is an integer from 0 to 5;
R' is a straight or branched alkyl of 1 to 5 carbon atoms
optionally substituted by one or two groups of --OH, alkoxy of 1 to
4 carbon atoms, thioalkoxy of 1 to 4 carbon atoms;
[0022] Sp is a straight or branched-chain divalent or trivalent
alkyl radical of 1 to 18 carbon atoms, divalent or trivalent aryl
or heteroaryl radical, divalent or trivalent cycloalkyl of 3 to 18
carbon atoms or heterocycloalkyl radical, divalent or trivalent
aryl- or heteroaryl-alkyl (C.sub.1-C.sub.18) radical, divalent or
trivalent cycloalkyl- or heterocyclo-alkyl-alkyl (C.sub.1-C.sub.18)
radical or divalent or trivalent unsaturated alkyl radical of 2 to
18 carbon atoms, wherein heteroaryl is furyl, thienyl,
N-methylpyrrolyl, pyridinyl, N-methylimidazolyl, oxazolyl,
pyrimidinyl, quinolyl, isoquinolyl, N-methylcarbazoyl,
aminocoumarinyl, or phenazinyl and wherein if Sp is a trivalent
radical, it can be additionally substituted by dialkylamino of 1 to
5 carbon atoms, alkoxy of 1 to 5 carbon atoms, hydroxy, or
alkylthio of 1 to 5 carbon atoms groups;
W' is
##STR00017##
[0023] R.sub.5 is --CH.sub.3, --C.sub.2H.sub.5, or
--CH(CH.sub.3).sub.2;
X is an iodine or bromine atom;
[0024] R.sub.5' is a hydrogen or the group RCO, wherein R is
hydrogen, branched or unbranched alkyl of 1 to 10 carbon atoms,
alkylene of 2 to 10 carbon atoms, aryl of 6 to 11 carbon atoms, a
(C.sub.6-C.sub.11) aryl-alkyl (C.sub.1-C.sub.5) group, or a
heteroaryl or heteroaryl-alkyl (C.sub.1-C.sub.5) group wherein
heteroaryl is defined as 2- or 3-furyl, 2- or 3-thienyl, 2- or
3-(N-methylpyrrolyl), 2-, 3-, or 4-pyridinyl, 2-, 4-, or
5-(N-methylimidazolyl), 2-, 4-, or 5-oxazolyl, 2-, 3-, 5-, or
6-pyrimidinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, or 1-, 3-,
4-, 5-, 6-, 7-, or 8-isoquinolyl, all aryl and heteroaryl groups
optionally substituted by one or more hydroxy, amino, carboxy,
halo, nitro, (C.sub.1-C.sub.3) alkoxy of 1 to 3 carbon atoms, or
thioalkoxy of 1 to 5 carbon atoms; and
Q is selected from the group consisting of --NNHCO--, --NNHCS--,
--NNHCONH--, --NNHCSNH--, and --NO--;
[0025] comprising the steps of: a. reacting a carboxylic acid of
the formula
##STR00018##
with a mercapto compound of the formula
H.sub.2Q-Sp-SH
in an alcohol solvent in the presence of an alkyl carboxylic acid,
alk.sup.2CO.sub.2H where alk.sup.2 is 1 to 4 carbon atoms at about
20.degree. to 70.degree. C. for about 1 to 24 hours, wherein
Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, and Sp are as defined above,
to produce a bilinker-carboxylic acid of the formula,
##STR00019##
b. isolating the bilinker-carboxylic acid of step (a); c. reacting
the isolated bilinker-carboxylic acid of step (b) with an at least
of N-hydroxysuccinimide, 2,3,5,6-tetrafluorophenol,
pentafluorophenol, 4-nitrophenol, 2,4-dinitrophenol, or
N-hydroxysulfosuccinimide in the presence of
1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or
N,N'-disuccinimidyl carbonate in an inert solvent containing 0-50%
N,N-dimethylformamide (DMF) to generate a trilinker-activated ester
of the formula
##STR00020##
where Z is hereinbefore defined d. reacting the trilinker-activated
ester in step c in the presence of a base or an organic base with
CH.sub.3--S--S--S--W' in an inert organic solvent to generate an
activated ester of the formula
##STR00021##
e. isolating the activated ester of step (d) and purifying to yield
compounds of Formula (I)
##STR00022##
[0026] The processes include a purifying of step comprising the use
of a reverse phase high performance liquid chromatography having a
mobile phase of about pH 7.0 to 9.0 followed with a normal phase
chromatography.
[0027] The processes include:
alk.sup.1 is 3 carbon atoms;
Sp.sup.1 is --O--;
Z.sup.1 is methyl;
Ar is unsubstituted 1,4-phenylene;
Sp is 4 carbon atoms;
R.sub.3 is H;
Q is NNHC(O)--;
##STR00023##
[0028] R.sub.5 is C.sub.2H.sub.5;
R.sub.5' is --C(O)--R;
R is methyl; and
Z is
##STR00024##
[0030] The processes include but are not limited to Alk.sup.1 is an
alkylene of 2 to 5 carbon atoms, and Sp.sup.1 is an oxygen
atom.
[0031] The processes include but are not limited Alk.sup.1 is an
alkylene of 3 carbon atoms.
[0032] The processes include but are not limited Z.sup.1 is alkyl
of 1 to 3 carbon atoms.
[0033] The processes include but are not limited Ar is 1,2-, 1,3-,
or 1,4-phenylene, or 1,2-, 1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-,
2,3-, 2,6-, or 2,7-naphthylidene.
[0034] The processes include but are not limited Ar is
1,4-phenylene.
[0035] The processes include but are not limited Q is
--NNHCO--.
[0036] The processes include but are not limited Sp is straight or
branched-chain divalent or trivalent alkyl radical of 1 to 12
carbon atoms.
[0037] The processes include but are not limited Sp is straight or
branched-chain divalent or trivalent alkyl radical of 1 to 6 carbon
atoms.
[0038] The processes include but are not limited the alcohol
solvent is methanol.
[0039] The processes include but are not limited the inert solvent
is acetonitrile.
[0040] The processes include but are not limited alkyl carboxylic
acid is acetic acid.
[0041] The processes include but are not limited the inert organic
solvent is acetonitrile.
[0042] The processes include but are not limited Z is
##STR00025##
[0043] Discussed herein is a process to prepare trilinker-activated
esters of the formula:
##STR00026##
wherein:
Z is selected from the group consisting of
##STR00027##
[0044] Alk.sup.1 is a branched or unbranched alkylene chain of 2 to
6 carbon atoms;
Sp.sup.1 is selected from --S--, --O--, --CONH--, --NHCO--, and
--NR'--;
Z.sup.1 is H, or alkyl of 1 to 5 carbon atoms;
[0045] Ar is 1,2-, 1,3-, or 1,4-phenylene optionally substituted
with one, two, or three groups independently selected from alkyl of
1 to 6 carbon atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1
to 4 carbon atoms, halogen, nitro, --COOR', --CONHR',
--O(CH.sub.2).sub.nCOOR', --S(CH.sub.2).sub.nCOOR',
--O(CH.sub.2).sub.nCONHR', and --S(CH.sub.2).sub.nCONHR' or a 1,2-,
1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 2,3-, 2,6-, or
2,7-naphthylidene optionally substituted with one, two, three, or
four groups independently selected from alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1 to 4 carbon
atoms, halogen, nitro, --COOR', --CONHR', --O(CH.sub.2).sub.nCOOR',
--S(CH.sub.2).sub.nCOOR', --O(CH.sub.2).sub.nCONHR', and
--S(CH.sub.2).sub.nCONHR'; n is an integer from 0 to 5;
R' is a straight or branched alkyl of 1 to 5 carbon atoms
optionally substituted by one or two groups of --OH, alkoxy of 1 to
4 carbon atoms, thioalkoxy of 1 to 4 carbon atoms;
[0046] Sp is a straight or branched-chain divalent or trivalent
alkyl radical of 1 to 18 carbon atoms, divalent or trivalent aryl
or heteroaryl radical, divalent or trivalent cycloalkyl of 3 to 18
carbon atoms or heterocycloalkyl radical, divalent or trivalent
aryl- or heteroaryl-alkyl (C.sub.1-C.sub.18) radical, divalent or
trivalent cycloalkyl- or heterocyclo-alkyl-alkyl (C.sub.1-C.sub.18)
radical or divalent or trivalent unsaturated alkyl radical of 2 to
18 carbon atoms, wherein heteroaryl is furyl, thienyl,
N-methylpyrrolyl, pyridinyl, N-methylimidazolyl, oxazolyl,
pyrimidinyl, quinolyl, isoquinolyl, N-methylcarbazoyl,
aminocoumarinyl, or phenazinyl and wherein if Sp is a trivalent
radical, it can be additionally substituted by dialkylamino of 1 to
5 carbon atoms, alkoxy of 1 to 5 carbon atoms, hydroxy, or
alkylthio of 1 to 5 carbon atoms groups;
W' is
##STR00028##
[0047] R.sub.5 is --CH.sub.3, --C.sub.2H.sub.5, or
--CH(CH.sub.3).sub.2;
X is an iodine or bromine atom;
[0048] R.sub.5' is a hydrogen or the group RCO, wherein R is
hydrogen, branched or unbranched alkyl of 1 to 10 carbon atoms,
alkylene of 2 to 10 carbon atoms, aryl of 6 to 11 carbon atoms, a
(C.sub.6-C.sub.11) aryl-alkyl (C.sub.1-C.sub.5) group, or a
heteroaryl or heteroaryl-alkyl (C.sub.1-C.sub.5) group wherein
heteroaryl is defined as 2- or 3-furyl, 2- or 3-thienyl, 2- or
3-(N-methylpyrrolyl), 2-, 3-, or 4-pyridinyl, 2-, 4-, or
5-(N-methylimidazolyl), 2-, 4-, or 5-oxazolyl, 2-, 3-, 5-, or
6-pyrimidinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, or 1-, 3-,
4-, 5-, 6-, 7-, or 8-isoquinolyl, all aryl and heteroaryl groups
optionally substituted by one or more hydroxy, amino, carboxy,
halo, nitro, (C.sub.1-C.sub.3) alkoxy of 1 to 3 carbon atoms, or
thioalkoxy of 1 to 5 carbon atoms; and
Q is selected from the group consisting of --NNHCO--, --NNHCS--,
--NHNCONH--, --NNCSNH--, and --NO--;
[0049] comprising the steps of: a. reacting a carboxylic acid of
the formula
##STR00029##
with a mercapto compound of the formula
H.sub.2Q-Sp-SH
in an alcohol solvent in the presence of an alkyl carboxylic acid,
alk.sup.2CO.sub.2H where alk.sup.2 is 1 to 4 carbon atoms at about
20.degree. to 70.degree. C. for about 1 to 24 hours, wherein
Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, and Sp are as defined above,
to produce a bilinker-carboxylic acid of the formula
##STR00030##
b. isolating the bilinker-carboxylic acid of step (a); c. reacting
the bilinker-carboxylic acid from step (b) in the presence of a
base or an organic base with CH.sub.3--S--S--S--W' in an inert
organic solvent to generate a bilinker-methyltrithio compound of
the formula
##STR00031##
d. reacting the isolated bilinker-methyltrithio compound of step
(c) with N-hydroxysuccinimide, 2,3,5,6-tetrafluorophenol,
pentafluorophenol, 4-nitrophenol, 2,4-dinitrophenol, or
N-hydroxysulfosuccinimide in the presence of
1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or
N,N'-disuccinimidyl carbonate in an inert solvent containing 0-50%
N,N-dimethylformamide (DMF) to generate a trilinker-activated ester
of the formula
##STR00032##
[0050] Presented herein is a process for the preparation of
trifunctional linker intermediates, of the formula
##STR00033##
wherein:
Alk.sup.1 is a branched or unbranched alkylene chain of 2 to 6
carbon atoms;
Sp.sup.1 is selected from --S--, --O--, --CONH--, --NHCO--, and
--NR'--;
Z.sup.1 is H, or alkyl of 1 to 5 carbon atoms;
[0051] Ar is 1,2-, 1,3-, or 1,4-phenylene optionally substituted
with one, two, or three groups independently selected from alkyl of
1 to 6 carbon atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1
to 4 carbon atoms, halogen, nitro, --COOR', --CONHR',
--O(CH.sub.2).sub.nCOOR', --S(CH.sub.2).sub.nCOOR',
--O(CH.sub.2).sub.nCONHR', and --S(CH.sub.2).sub.nCONHR' or a 1,2-,
1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 2,3-, 2,6-, or
2,7-naphthylidene optionally substituted with one, two, three, or
four groups independently selected from alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 5 carbon atoms, thioalkoxy of 1 to 4 carbon
atoms, halogen, nitro, --COOR', --CONHR', --O(CH.sub.2).sub.nCOOR',
--S(CH.sub.2).sub.nCOOR', --O(CH.sub.2).sub.nCONHR', and
--S(CH.sub.2).sub.nCONHR'; n is an integer from 0 to 5;
R' is a straight or branched alkyl of 1 to 5 carbon atoms
optionally substituted by one or two groups of --OH, alkoxy of 1 to
4 carbon atoms, thioalkoxy of 1 to 4 carbon atoms;
[0052] Sp is a straight or branched-chain divalent or trivalent
alkyl radical of 1 to 18 carbon atoms, divalent or trivalent aryl
or heteroaryl radical, divalent or trivalent cycloalkyl of 3 to 18
carbon atoms or heterocycloalkyl radical, divalent or trivalent
aryl- or heteroaryl-alkyl (C.sub.1-C.sub.18) radical, divalent or
trivalent cycloalkyl- or heterocyclo-alkyl-alkyl (C.sub.1-C.sub.18)
radical or divalent or trivalent unsaturated alkyl radical of 2 to
18 carbon atoms, wherein heteroaryl is furyl, thienyl,
N-methylpyrrolyl, pyridinyl, N-methylimidazolyl, oxazolyl,
pyrimidinyl, quinolyl, isoquinolyl, N-methylcarbazoyl,
aminocoumarinyl, or phenazinyl and wherein if Sp is a trivalent
radical, it can be additionally substituted by dialkylamino of 1 to
5 carbon atoms, alkoxy of 1 to 5 carbon atoms, hydroxy, or
alkylthio of 1 to 5 carbon atoms groups;
Q is selected from the group consisting of --NNHCO--, --NNHCS--,
and --NNHCONH--;
Z is selected from the group consisting of
##STR00034##
[0053] comprising the steps of: a. reacting a carboxylic acid of
the formula
##STR00035##
with a mercapto compound of the formula
H.sub.2Q-Sp-SH
in an alcohol solvent in the presence of an alkyl carboxylic acid,
alk.sup.2CO.sub.2H, where alk.sup.2 is 1 to 4 carbon atoms at about
20.degree. to 70.degree. C. for about 1 to 24 hours, wherein
Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, and Sp are as defined above,
to produce a bilinker-carboxylic acid of the formula
##STR00036##
b. isolating the bilinker-carboxylic acid of step (a); c. reacting
the isolated bilinker-carboxylic acid of step (b) with
N-hydroxysuccinimide, 2,3,5,6-tetrafluorophenol, pentafluorophenol,
4-nitrophenol, 2,4-dinitrophenol, or N-hydroxysulfosuccinimide in
the presence of 1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or
N,N'-disuccinimidyl carbonate in an inert solvent containing 0-50%
DMF to generate trifunctional linker intermediates, of the
formula
##STR00037##
[0054] The terms used in this specification generally have their
ordinary meanings in the art, within the context of the invention,
and in the specific context where each term is used. Certain terms
are discussed below, or elsewhere in the specification, to provide
additional guidance to the practitioner in describing the
compounds, compositions, and methods of the invention and how to
make and use them. Moreover, it will be appreciated that the same
thing can be said in more than one way. Consequently, alternative
language and synonyms may be used for any one or more of the terms
discussed herein, nor is any special significance to be placed upon
whether or not a term is elaborated or discussed herein. The use of
examples anywhere in this specification, including examples of any
terms discussed herein, is illustrative only, and in no way limits
the scope and meaning of the invention or of any exemplified term.
Likewise, the invention is not limited to the examples
presented.
[0055] The terms, "about" or "approximately" shall generally mean
within 20 percent, preferably within 10 percent, and more
preferably within 5 percent of a given value or range.
[0056] The term "alkoxy" as used herein refers to an alkyl group,
as defined above, having an oxygen radical attached thereto. The
term "thioalkoxy" refers to an alkoxy group as defined, having a
sulfur radical attached thereto.
[0057] The term "alkyl" refers to the radical of saturated
aliphatic groups, including straight-chain alkyl groups,
branched-chain alkyl groups, cycloalkyl(alicyclic) groups, alkyl
substituted cycloalkyl groups, and cycloalkyl substituted alkyl
groups. In an embodiment, a straight chain or branched chain alkyl
has 6 or fewer carbon atoms in its backbone. The term "alkyl" can
be used alone or as part of a chemical name. The term "aryl" is
defined as an aromatic carbocyclic moiety and may be substituted or
unsubstituted. Aryl groups have 6 to 14 carbon atoms and include
phenyl and napthyl. The term "aryl" also includes polycyclic ring
systems having two or more rings in which two or more carbons are
common to two adjoining rings (the rings are "fused") wherein at
least one of the carbocyclic rings is aromatic, e.g., the other
rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, and/or
aryls.
[0058] Carboxy is defined as a --CO.sub.2H radical.
[0059] The term "halogen" or "halo" refers to an atom of fluorine,
chlorine, bromine, or iodine.
[0060] The term "heteroaryl" refers to a 4 to 10 membered ring
structure, which ring structure includes one to four heteroatoms. A
heteroaryl comprises a heterocyclic ring system of one to three
fused rings, in which at least one ring may have an aromatic
character and contains 1 to 4 heteroatoms the same or different
selected from the group consisting of S, N, and O. The remaining
rings of the ring system may be fully unsaturated, partially
saturated, or fully saturated. Each ring comprises three to ten
members. The term "heteroatom" as used herein means an atom of any
element other than carbon or hydrogen and include for example
nitrogen, oxygen, sulfur, phosphorus, and selenium.
[0061] The term "nitro" means --NO.sub.2.
[0062] The term "substituted" is contemplated to include all
permissible substituents of organic compounds. In a broad aspect,
the permissible substituents of organic compounds include acyclic
and cyclic, branched and unbranched, carbocyclic and heterocyclic,
aromatic and nonaromatic substituents of organic compounds. The
permissible substituents can be one or more and the same or
different for appropriate organic compounds. For purposes of this
invention, the heteroatoms such as nitrogen may have hydrogen
substituents and/or any permissible substituents of organic
compounds described herein which satisfy the valencies of the
heteroatoms. This invention is not intended to be limited in any
manner by the permissible substituents of organic compounds.
[0063] The term alkyl means a straight or branched alkyl of 1 to 18
carbon atoms, preferably 2 to 5 carbon atoms, 1 to 4 carbon atoms
or 1 to 3 carbon atoms. In some embodiments of the invention the
term alkyl is methyl.
[0064] The term alcohol solvent means methanol, ethanol and the
like with a boiling point of less than about 100.degree. C.
[0065] Alkylene refers to an alkyl, as defined above, which is a
diradical, rather than a radical. An alkylene can be branched or
unbranched and can have 2-18 carbons.
[0066] Ambient temperature is about 25.degree. C.
[0067] Inert solvent or inert organic solvent describes a solvent
that will neither react or form a covalent bond in the steps to
prepare compounds of Formula (I), trilinker-activated esters or
trifunctional linker intermediates as described herein. An example
of an inert solvent or inert organic solvent may be mixtures such
as but not limited to acetonitrile/ethylacetate (1:1)),
dichloromethane, N,N-dimethylformamide, tetrahydrofuran, dioxane or
acetonitrile. An inert solvent may contain 0-50%
N,N-dimethylformamide. An inert organic solvent is for example
acetonitrile, ethyl acetate or dichloromethane.
[0068] An organic base is for example an alkylamine base which
includes triethylamine, N,N-diethylmethylamine, N,N-diethylaniline,
N,N-diethylethylenediamine, N,N-diisopropylethylamine,
tributylamine or tripropylamine and further include
dimethylaminopyridine (DMAP) with diisopropylethylamine (DIEA),
N-methylmorpholine, N-methylpyrrolidine,
2,6-di-tertbutyl-4-methylpyridine or pyridine. A base is an alkali
metal hydroxide, alkali metal carbonate or alkali metal
bicarbonate.
DETAILED DESCRIPTION OF THE INVENTION
[0069] The process for the preparation of calicheamicin derivatives
of Formula I and tri and bifunctional linker intermediates useful
in the preparation of said derivatives of the present invention are
described in the following reaction Schemes I and II.
##STR00038##
[0070] In accordance with Scheme I, a carboxylic acid 1 wherein
Alk.sup.1, Sp.sup.1, Ar and Z.sup.1 are hereinbefore defined, found
in U.S. Pat. No. 5,773,001, which is hereby incorporated herein by
reference, are condensed with mercapto compound 2 where Sp and Q
are hereinbefore defined in an alcoholic solvent with a boiling
point of less than about 100.degree. C. in the presence of an alkyl
carboxylic acid in a about 5% acetic acid at about 20.degree. to
about 70.degree. C. for about 1 to about 24 hours, to afford
bilinker-carboxylic acid 3 wherein Alk.sup.1, Sp.sup.1, Ar, Q, Sp
and Z.sup.1 are as defined above.
[0071] Bilinker-carboxylic acid 3 is reacted with
N-hydroxysuccinimide, 2,3,5,6-tetrafluorophenol, pentafluorophenol,
4-nitrophenol, 2,4-dinitrophenol, or N-hydroxysulfosuccinimide in
the presence of 1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or other
carbodiimide or N,N'-disuccinimidyl carbonate in an inert solvent
such as dichloromethane, tetrahydrofuran, dioxane, or acetonitrile
containing 0-50% DMF or DMF to generate trilinker-activated ester 4
where Z is selected from the group consisting of
##STR00039##
[0072] For example, reaction of bilinker-carboxylic acid 3 with a
coupling agent, such as 1,3-dicyclohexylcarbodiimide (DCC) or
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, and
N-hydroxysuccinimide or other comparable carboxyl-activating group
in an inert solvent, such as N,N-dimethylformamide (DMF),
tetrahydrofuran, dioxane or acetonitrile, leads to the formation of
a trilinker-activated ester 4, such as the N-hydroxysuccinimide
ester described herein. Preferred is N-hydroxysuccinimide, DCC at
ambient temperature in dioxane. A preferred solvent mixture is
acetonitrile containing 0-50% DMF. Reaction of the
bilinker-carboxylic acid 3 with N-hydroxysuccinimide,
2,3,5,6-tetrafluorophenol, pentafluorophenol, 4-nitrophenol,
2,4-dinitrophenol, or N-hydroxysulfosuccinimide in the presence of
1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or other
carbodiimide in an inert solvent such as dioxane or acetonitrile
containing 0-50% N,N-dimethylformamide (DMF) leads to the formation
of a trilinker-activated ester 4. The trilinker-activated ester 4
can be isolated by removal of the volatile solvents and further
purified by reverse or normal phase chromatography on an inert
support which includes silica-60.
[0073] Trilinker-activated ester 4 is first reacted with an alkali
metal carbonate which includes but is not limited to sodium
carbonate and forms the sodium salt of trilinker-activated ester 4
in acetonitrile by heating at gentle reflux. Further reaction of
the sodium salt of trilinker-activated ester 4 with
methyltrithioantitumor antibiotic 5 at about -15.degree. C. in an
inert organic solvent, preferably acetonitrile gives activated
ester 6 wherein Z, Alk.sup.1, Sp.sup.1, Ar, Z.sup.1, Q, Sp and W'
are hereinbefore defined. In particular, N-acetyl-LL-E33288
.gamma..sub.1.sup.l is the preferred methyltrithioantitumor
antibiotic 5. Preferred is the reaction in acetonitrile at about
0.degree. C. Optionally an organic base may replace the alkali
metal carbonate which preferably includes triethylamine, in
acetonitrile at about 0.degree. C.
[0074] As further described in Scheme II, reaction of
bilinker-carboxylic 3 prepared by condensation of carboxylic acid 1
wherein Alk.sup.1, Sp.sup.1, Ar and Z.sup.1 are hereinbefore
defined, with mercapto compound 2 according to scheme I, is reacted
with methyltrithioantitumor antibiotic 5 in the presence of
triethylamine in N,N-dimethylformamide (DMF) at about -5.degree. C.
affords intermediate 7 which is further converted to
trilinker-activated ester 6 by reaction with N-hydroxysuccinimide,
2,3,5,6-tetrafluorophenol, pentafluorophenol, 4-nitrophenol,
2,4-dinitrophenol, or N-hydroxysulfosuccinimide in the presence of
1,3-dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), or other
carbodiimide or N,N'-disuccinimidyl carbonate in an inert solvent
mixture of DMF and acetonitrile which is then purified preferably
by chromatography to afford antitumor antibiotics of Formula
(I).
##STR00040##
[0075] The following examples are presented to illustrate certain
embodiments of the present invention, but should not be construed
as limiting the scope of this invention. Those skilled in the art
will readily understand that known variations of the conditions of
the following preparative procedures can be used to prepare these
compounds.
EXAMPLE 1
Butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-(4-hydroxy-4-oxobutoxy)phenyl]ethylidene-
]hydrazide
##STR00041##
[0077] To a stirred mixture of 0.5 g [3.4 mmol] of
3-methyl-3-mercapto-butanoic acid hydrazide in 5.0 ml of methanol
is rapidly added 0.91 g [4.1 mmol] of 4-(4-acetylphenoxy)-butanoic
acid followed by an additional 10 ml of methanol and 1.5 ml of
acetic acid and stirring continued for 24 hours. The reaction
mixture is filtered and the solid washed with 100 ml of methanol to
give 0.78 g of the title compound as a solid.
EXAMPLE 2
Butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-(4-hydroxy-4-oxobutoxy)phenyl]ethylidene-
]hydrazide
##STR00042##
[0079] A mixture of 3-methyl-3-mercapto-butanoic acid hydrazide
(4.0 g, 27 mmol), 4-(4-acetylphenoxy)-butanoic acid (5.0 g, 22.5
mmol) and acetic acid (7.5 mL) in methyl alcohol (75 mL) is heated
at about 45.degree. C. for about 7 h. The mixture is allowed to
cool to room temperature. The white solid (7.12 g, 90%) is
collected on a Buchner funnel and washed with MeOH (2.times.10 mL).
.sup.1H NMR (DMSO-d.sub.6): .delta. (ppm) 12.14 (s, 1H), 10.37 and
10.21 (s, 1H), 7.74-7.70 (m, 2H), 6.97-6.95 (m, 2H), 4.04 (t, 2H),
3.09 and 3.04 (s, 2H), 2.66 (s, 1H), 2.41-2.37 (t, 2H), 2.22 and
2.20 (s, 3H), 1.97-1.93 (m, 2H), 1.8 (s, 3H), 1.47 (s, 3H). MS: 375
(M.sup.++Na).
EXAMPLE 3
Butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-[4-[(2,5-dioxo-1-pyrrolidinyl)oxy]-4-oxo-
butoxy]phenyl]ethylidene]hydrazide
##STR00043##
[0081] To a mixture of 0.5 g [1.42 mmol] of butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-(4-hydroxy-4-oxobutoxy)phenyl]ethylidene-
]hydrazide (Example 1 or 2) and 0.22 g [1.89 mmol] of
N-hydroxysuccinimide is added 10 ml of N,N-dimethylformamide
followed by the rapid addition of 0.70 g [3.65 mmol] of
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and the
mixture stirred at room temperature for 3 hours. The reaction
mixture is concentrated in vacuo to a residue which is partitioned
between ethyl acetate and water. The separated organic layer is
washed with water, saturated sodium chloride and dried
(MgSO.sub.4). The organic layer is evaporated in vacuo to give an
oily residue which crystallized from ethyl acetate-hexane affording
0.21 g of the title compound as a colorless solid.
EXAMPLE 4
Butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-[4-[(2,5-dioxo-1-pyrrolidinyl)oxy]-4-oxo-
butoxy]phenyl]ethylidene]hydrazide
##STR00044##
[0083] A mixture of butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-(4-hydroxy-4-oxobutoxy)phenyl]ethylidene-
]hydrazide (Examples 1 or 2) (3.69 g, 10.48 mmol) and
N-hydroxysuccinimide (1.386 g, 12.05 mmol) is suspended in dioxane
(60 mL), DCC (2.482 g, 12.05 mmol) in dioxane (30 mL) is added
dropwise over 15 min. The mixture is stirred at room temperature
for 24 h. The precipitated dicyclohexylurea is filtered off and
washed with dioxane (2.times.10 mL). The filtrate is concentrated
to about 50 mL and while stirring, water (250 mL) is added. The
resulting white solid is collected on a Buchner funnel, washed with
water (2.times.50 mL), and dried in vacuo at room temperature. To
this white solid is added MeCN (60 mL) and the mixture is heated at
50.degree. C. until it became solution, isopropyl alcohol (IPA)
(400 mL) is added. The mixture is then cooled to 0-5.degree. C. for
2 h. The solid is collected on a Buchner funnel, washed with cold
IPA (2.times.20 mL) an dried in vacuo to afford the product of the
example as a white solid (3.58 g,.sub.--70%). MS: 450
(M.sup.++1).
EXAMPLE 5
Butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-[4-[(2,5-dioxo-1-pyrrolidinyl)oxy]-4-oxo-
butoxy]phenyl]ethylidene]hydrazide condensed with
N-acetyl-LL-E33288.gamma..sub.1.sup.l
##STR00045##
[0085] To a stirred solution of 0.505 g [1.12 mmol] butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-[4-[(2,5-dioxo-1-pyrrolidinyl)oxy]-4-oxo-
butoxy]phenyl]ethylidene]hydrazide (Example 4) in 50 ml of
acetonitrile is added 0.123 g [1.16 mmol] of sodium carbonate
followed by heating at gentle reflux for 1 hour, cooled to room
temperature and filtered. The filtrate is cooled to -15.degree. C.
and a solution of 1.4969 g [1.1 mmol] of N-acetyl-LL-E33288
.gamma..sub.1.sup.l in 5 ml of acetonitrile added slowly by
dropwise addition over 20 minutes and stirring continued for about
1.5 hours. The reaction mixture is allowed to warm to room
temperature and stirred for about 3 hours. The volatiles are
evaporated in vacuo to a residue which is stored in a freezer. To
the residue is added 25 ml of ethyl acetate followed by storage in
a freezer for about 1 hour. The reaction mixture is filtered and
the ethyl acetate evaporated to a residue which is dissolved in 10
ml of ethyl acetate and applied to a column of 110 g of silica gel.
The column is eluted with 1-5% methyl alcohol in ethyl acetate to
give 0.322 g of the desired product having 65.27% purity as
determined by HPLC.
EXAMPLE 6
Butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-[4-[(2,5-dioxo-1-pyrrolidinyl)oxy]-4-oxo-
butoxy]phenyl]ethylidene]hydrazide condensed with
N-acetyl-LL-E33288.gamma..sub.1.sup.l
##STR00046##
[0087] A solution of butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-[4-[(2,5-dioxo-1-pyrrolidinyl)oxy]-4-oxo-
butoxy]phenyl]ethylidene]hydrazide (450 mg, 1 mmol) (Example 4) in
CH.sub.3CN (100 mL) containing Et.sub.3N (0.35 mL) is treated with
a solution of N-acetyl-LL-E33288 .gamma..sub.1.sup.l (500 mg, 0.355
mmol) in CH.sub.3CN (100 mL) at 0-5.degree. C. The mixture is then
stirred for another 1 h while cooling with a ice-bath. The solvent
is removed and the residue is purified on a silica gel column
eluting with CH.sub.2Cl.sub.2-MeOH to afford the product of the
example (340 mg, 54%) as a white solid. MS: 1780 (M.sup.++1)
EXAMPLE 7
Butanoic acid,
3-mercapto-3-methyl-,2[(E)-1-[4-(4-hydroxy-4-oxobutoxy)phenyl]ethylidene]-
hydrazide condensed with N-acetyl-LL-E33288.gamma..sub.1.sup.l
##STR00047##
[0089] To a stirring solution of N-acetyl-LL-E33288
.gamma..sub.1.sup.l (200 mg, 0.142 mmol) in 10 ml
acetonitrile/ethyl acetate (1:1) at -5.degree. C. is added in 1 ml
aliquots every 10 min a solution of butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-(4-hydroxy-4-oxobutoxy)phenyl]ethylidene-
]hydrazide (150 mg, 0.43 mmol) (Examples 1 or 2) in 10 ml
acetonitrile/ethyl acetate (1:1) and 0.06 ml triethyl amine. The
solution is stirred for two hours at -5.degree. C. after the last
addition of the butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-(4-hydroxy-4-oxobutoxy)phenyl]ethylidene-
]hydrazide solution. The solvent was removed under reduced pressure
and the residue is purified on a silica gel column eluting with
CH.sub.2Cl.sub.2-MeOH to afford the product of the example as a
white solid. MS 1684 (M.sup.++1)
EXAMPLE 8
Butanoic acid,
3-mercapto-3-methyl-,2-[(E)-1-[4-[4-[(2,5-dioxo-1-pyrrolidinyl)oxy]-4-oxo-
butoxy]phenyl]ethylidene]hydrazide condensed with
N-acetyl-LL-E33288.gamma..sub.1.sup.l
[0090] To a stirring solution of butanoic acid,
3-mercapto-3-methyl-,2[(E)-1-[4-(4-hydroxy-4-oxobutoxy)phenyl]ethylidene]-
hydrazide with N-acetyl-LL-E33288.gamma..sub.1.sup.l (100 mg, 0.059
mmol) in 0.5 mL of DMF and 1.8 mL of acetonitrile at 25.degree. C.
is added N-hydroxysuccinimide (236 mg, 2.05 mmol) and
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (160 mg 0.835 mmol).
Following the addition, the solution is stirred for one hour at
25.degree. C. The acetonitrile is removed under reduced pressure
and the resulting DMF solution is added to 3 mL of stirring water
giving a precipitate. The precipitate is filtered, dried and
purified on a silica gel column eluting with
CH.sub.2Cl.sub.2-isopropyl alcohol giving the product of the
example (53 mg, 50%) obtained as a white solid. MS: 1780
(M.sup.++1)
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