CA2157412A1 - Balanoids - Google Patents
BalanoidsInfo
- Publication number
- CA2157412A1 CA2157412A1 CA002157412A CA2157412A CA2157412A1 CA 2157412 A1 CA2157412 A1 CA 2157412A1 CA 002157412 A CA002157412 A CA 002157412A CA 2157412 A CA2157412 A CA 2157412A CA 2157412 A1 CA2157412 A1 CA 2157412A1
- Authority
- CA
- Canada
- Prior art keywords
- hydroxy
- phenyl
- benzene
- dihydroxy
- hydroxyphenyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D257/00—Heterocyclic compounds containing rings having four nitrogen atoms as the only ring hetero atoms
- C07D257/02—Heterocyclic compounds containing rings having four nitrogen atoms as the only ring hetero atoms not condensed with other rings
- C07D257/04—Five-membered rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
- C07C235/42—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton
- C07C235/44—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring
- C07C235/54—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a ring other than a six-membered aromatic ring
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C255/00—Carboxylic acid nitriles
- C07C255/49—Carboxylic acid nitriles having cyano groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
- C07C255/52—Carboxylic acid nitriles having cyano groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton to carbon atoms of six-membered aromatic rings being part of condensed ring systems
-
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C311/00—Amides of sulfonic acids, i.e. compounds having singly-bound oxygen atoms of sulfo groups replaced by nitrogen atoms, not being part of nitro or nitroso groups
- C07C311/01—Sulfonamides having sulfur atoms of sulfonamide groups bound to acyclic carbon atoms
- C07C311/02—Sulfonamides having sulfur atoms of sulfonamide groups bound to acyclic carbon atoms of an acyclic saturated carbon skeleton
- C07C311/09—Sulfonamides having sulfur atoms of sulfonamide groups bound to acyclic carbon atoms of an acyclic saturated carbon skeleton the carbon skeleton being further substituted by at least two halogen atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D207/14—Nitrogen atoms not forming part of a nitro radical
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/46—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with hetero atoms directly attached to the ring nitrogen atom
- C07D207/48—Sulfur atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
- C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D211/56—Nitrogen atoms
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D223/00—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom
- C07D223/02—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom not condensed with other rings
- C07D223/06—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom not condensed with other rings with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D223/12—Nitrogen atoms not forming part of a nitro radical
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D313/00—Heterocyclic compounds containing rings of more than six members having one oxygen atom as the only ring hetero atom
- C07D313/02—Seven-membered rings
- C07D313/04—Seven-membered rings not condensed with other rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D337/00—Heterocyclic compounds containing rings of more than six members having one sulfur atom as the only ring hetero atom
- C07D337/02—Seven-membered rings
- C07D337/04—Seven-membered rings not condensed with other rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
- C07F9/02—Phosphorus compounds
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
- C07F9/553—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having one nitrogen atom as the only ring hetero atom
- C07F9/572—Five-membered rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2601/00—Systems containing only non-condensed rings
- C07C2601/06—Systems containing only non-condensed rings with a five-membered ring
- C07C2601/08—Systems containing only non-condensed rings with a five-membered ring the ring being saturated
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2601/00—Systems containing only non-condensed rings
- C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
- C07C2601/14—The ring being saturated
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2601/00—Systems containing only non-condensed rings
- C07C2601/18—Systems containing only non-condensed rings with a ring being at least seven-membered
Abstract
A novel class of therapeutic compounds, denominated Balanoids, is disclosed. Balanoids have protein kinase C inhibitory activity and selectivity among the isoforms of protein kinase C. Balanoids are useful for treatment of diseases related to protein kinase C in animals, specially humans and is especially indicated for treatment of inflammatory diseases.
Description
DEMANDES OU BREVETS VOLUMINEUX
LA PRESENTE PARTIE DE ~; I I t DENIANDE OU CE BREVET
COMPREND PLUS D'UN TOME.
CECI EST LE TOME I DE ;2 NO~: Pour les tomes additionels, veuillez c~ntacter le 8ureau canadien des brevets JUMBO APPLICATIONSIPATENTS
THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE
THAN ONE VOLlJME
THIS IS VOLUME -1 - OF ,~
NOTE: Fcr additicnal Yclumes please c~nlact the Canadian Patent Office
LA PRESENTE PARTIE DE ~; I I t DENIANDE OU CE BREVET
COMPREND PLUS D'UN TOME.
CECI EST LE TOME I DE ;2 NO~: Pour les tomes additionels, veuillez c~ntacter le 8ureau canadien des brevets JUMBO APPLICATIONSIPATENTS
THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE
THAN ONE VOLlJME
THIS IS VOLUME -1 - OF ,~
NOTE: Fcr additicnal Yclumes please c~nlact the Canadian Patent Office
2 1 5 ~ PCT/US94/02283 BALANOIDS
~ , FIELD OF THE ~Nv~ ON
The present invention relates to the field of treatments for inflammatory, cardiovascular, metabolic, nervous system, viral infectious, neoplastic and other diseases. The invention provides compounds which can inhibit protein kinase C enzymes. More particularly, the present invention relates to novel compounds which are referred to herein as "balanoids".
R~R~.~UND OF THE INVENTION
Protein kinase C (PRC) is a family of calcium- and phospholipid-dependent serine/threonine-specific protein kinases which play an important role in cellular growth control, regulation, and differentiation. Protein kinase C is activated by diacylglycerol (DAG), a neutral lipid, and when activated will transfer the y-phosphate of MgATP to a serine or threonine residue on a substrate protein. The mechanisms of protein kinase C action have been described in U.S. Patent 4,816,450 issued March 28, 1989 to Bell et al., which is incorporated herein by reference.
The activation of protein kinase C has been implicated in several human disease processes, including cancer tumors, inflammation and reperfusion injury. Accordingly, protein kinase C is a target for therapeutic agents useful in treating these conditions.
Cancer is a disease characterized in part by uncontrolled cell growth. Protein kinase C is directly involved in cellular growth control and is believed to be involved in tumor formation. Protein kinase C is fundamental W094/20062 PCT~S94/02283 _ 2 1 5 l ~ 1 2 -- 2 to the processes involved in tumorigenicity, since it is the major high-affinity receptor for endogenous cellular DAGs as well as for several classes of tumor promoters. These tumor promoters also stimulate protein kinase C catalysis. Castagna et al., (1982) J. Biol. Chem. 257:7847, reported direct activation of protein kinase C by tumor promoting phorbol esters.
Protein kinase C is the major, if not exclusive, intracellular receptor of phorbol esters, which are very potent tumor promoters. Phorbol esters and other tumor promoters bind to and activate protein kinase C. Since DAG and phorbol esters interact at the same site, DAGs have been suggested to be the "endogenous phorbol esters", analogous to the opiate receptor where the conservation of a high affinity receptor implied the existence of an endogenous analogue. DAG has been shown to increase the affinity of protein kinase C for Ca 2 and phospholipid and thus activates protein kinase C at cellular levels of these essential cofactors. Extracellular signals including hormones, growth factors, and neurotransmitters are known to stimulate phosphatidylinositol turnover resulting in the generation of IP3 and DAG.
Structures of 40 distinct oncogenes of viral and cellular origin have revealed that oncogenes encode altered forms of normal cellular proteins. Several of the gene products appear related to growth factors or other elements involved in transmembrane signalling. These oncogene products appear to function by altering the level of critical second messengers. Cells transformed with the oncogenes ras, sis, erbB, abl, and src have been shown to contain elevated levels of DAG which is then believed to activate protein kinase C.
Studies on ras transformed cells have shown protein kinase C
activation to be concomitant with elevation of DAG.
Phorbol esters, such as phorbol myristate acetate (PM~A), have complex effects on cells including effects on membrane function, mitogenesis, differentiation, and gene expression. Synthetic DAGs mimic many of the effects of PMA in vitro and inhibitors of protein kinase C have been shown to 215~412 W094/20062 PCT~S94/02283 block PMA-induced effects on cells. Thus, protein kinase C may mediate the actions of certain oncogenes, such as ras, which cause intracellular increases in DAG and concomitant increases in protein kinase C. In addition, activation of protein kinase C leads to the expression of c-myc, c-fos, c-cis, c-fms, nuclear protooncogenes which are important in cell transformation. Overexpression of protein kinase C in NIH 3T3 cells causes altered growth regulation and enhanced tumorige-nicity, and in rat fibroblasts leads to anchorage-independent growth in soft agar. Overexpression of protein kinase C in these cells resulted in tumor formation in animals receiving transplanted cells.
Several studies have shown increased expression of protein kinase C in certain tumor types such as breast and lung carcinomas. Activated protein kinase C has also been detected in human colon carcinomas although increased expression at the gene level was not seen. Topoisomerases are directly modulated by protein kinase C as substrates for the enzyme.
Protein kinase C inhibitors have been reported to potentiate the antitumor activity of chemotherapeutic agents such as cis-platin both in vitro and in vivo (Grunicke, et al.
(1989) Adv. Enzyme Regul. 28:201; and German Offenlegungs-schrift DE 3827974). In addition, it has been suggested that protein kinase C would be a potential target for therapeutic design because of its central role in cell growth (Tritton, T.R. and J.A. Hickman, (1990) Cancer Cells 2:5-102). German Offenlegungsschrift DE 3827974 Al discloses therapeutic preparations comprising a protein kinase C inhibitor in combination with a lipid, a lipid analogue, a cytostatic agent or phospholipase inhibitor which are useful for cancer therapy.
Inflammation and reperfusion injury, particularly pertaining to cardiac injury, are common conditions for which there exists no definitive treatment despite extensive research. Appropriate treatments for these conditions are needed.
Protein kinase C inhibitors have been demonstrated to block platelet aggregation and release of neutrophil activating W094/20062 PCT~S94/02283 ~15~12 - 4 ~
agents such as platelet activating factor (PAF) (Schachtele, et al. (1988) Biochem. Biophy. Res. Commun. 151:542; Hannun, et al. (1987) J. Biol. Chem. 262:13620; Yamada, et al. (1988) Biochem. Pharmacol. 37:1161). Protein kinase C inhibitors have also been shown to inhibit neutrophil activation, and chemotactic migration (McIntyre, et al. (1987) J. Biol Chem.
262:15730; Lambreth, et al. (1988) J. Biol. Chem. 263:3818;
Pittet, et al. (1987) J. Biol. Chem. 262:10072; and Gaudry, et al. (1988) Immunology 63:715), as well as neutrophil degranulation and release of proteolytic enzymes and reactive oxygen intermediates (Wilson, et al. (1986) J. Biol. Chem.
261:12616; Fujita et al. (1986) Biochem. Pharmacol. 35:4555;
Berkow, et al. (1987) J. Leukoc. Biol. 41: 441; Salzer, et al.
(1987) Biochem. Biophys. Res. Commun. 148:747; Kramer, et al.
(1989) J. Biol. Chem. 262:5876; and Dewald, et al. (1989) Biochem. J. 26~:879).
Thus, inhibitors of protein kinase C have the capability of blocking all three of the most significant mechanisms of pathogenesis associated with myocardial reperfusion injury. Protein kinase C is, accordingly, a drug target for therapeutic agents. Additionally, the inhibitory effect of protein kinAse C inhibitors on keratinocytes, and on the oxidative burst in neutrophils, provides an anti-inflammatory effect.
Groth, T., et al., Proc. Adabori Conf: 3rd Ger.-Jap.
Symp. Pept. Chem., E. Wuensch, ed., 91 (1989) disclose ophiocordin, an antibiotic with antifungal activity having certain structural similarities to certain compounds of the invention.
~UNMARY OF ~HE l~v~.,lON
The present invention relates to a novel class of compounds referred to herein as balanoids. Compounds according to the present invention have the following formula:
2157~12 W O 94/20062 PCTrUS94102283 G F E
1' D l~ B B Z
~ ~ 1 2 n(~ ~)m A
wherein:
A is: CH2, NR, S, S 2 or O;
Bl is : NR, O or CH2;
B2 is: C0, CS, or SO2;
Z is: R4, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
D is: NR3, O or CH2;
E is: R5, aryl, heteroaryl, substituted aryl or substituted heteroaryl:
F is : CO , CS , CH (OR6), CH2 , O , S or NR;
G is: R7, aryl, heteroaryl, substituted aryl, substituted heteroaryl or substituted cycloalkyl;
K is: hydrogen or lower alkyl;
X is: CO, CS, CH2, CNR or CCR R
R , R , R , R , R , R , R , R and Rl are, independently, hydrogen, lower alkyl, aryl or JR ;
R5 is: lower alkyl or aryl;
J is: CO, C=NR, S 2 or P(O)0 alkyl;
Rll is: hydrogen, lower alkyl, aryl, alkamino, arylamino, aryloxy or alkoxy;
Rl2 is: straight or branched alkyl, aryl;
m is: 1-4;
n is: 1-4; and m plus n is up to 5;
providing that if m is 3, A is NH, Bl is 0, B2 is CO, Z is p-hydroxyphenyl, D is NH, X is CO, and E, F, and G, taken together, are W094/20062 PCT~S94/02283 215~12 - 6 -OH HO
HOOC ~$ C
OH
then n is not 1.
Additionally, the present invention relates to pharma-ceutically acceptable salts of the above compounds and to formulations comprising the above compounds in pharmaceutically acceptable carriers. Prodrugs such as carbonates and esters of phenolic functional groups and other species metabolizable into compounds of the invention are also considered to be within the scope of the present invention.
The present invention relates to a method of inhibiting protein kinase C activity which comprises contacting protein kinase C with an inhibitory amount of a balanoid of the invention.
The present invention also relates to methods of treating an animal, preferably a mammal, that is suffering from a PRC-related disease, especially an inflammatory, cardiovascular and/or neoplastic diseases by administering an effective amount of a balanoid to the animal.
DET~TT-~n ~ TPTION OF ~K~KK~ EMBODINENT8 This invention is directed to a family of novel compounds denominated "balanoids". Members of the family have been found to exhibit the ability to inhibit enzymes of the family of enzymes known as protein kinase C enzymes.
Selectivity in inhibitors among the isoforms of protein kinase C (PKC) has been shown for balanoids and it is believed that balanoids will be useful in the treatment of disease linked to PKC enzymes.
Methods for preparing balanoids together with synthetic intermediates, are further objects of the invention as are methods for testing for PKC-linked diseases.
W094/20062 215 ~ PCT~S94102283 The present invention relates to balanoids, and their pharmaceutically acceptable salts and formulations. Compounds according to the present invention have been shown to inhibit protein kinase C. PKC inhibitors are known to be useful in the treatment of cancer, inflammatory and reperfusion injury through their antiproliferative and anti-inflammatory activities in human neutrophils, human keratinocytes, and human tumor cells.
The present invention relates to methods of inhibiting protein kinase C activity which comprises contacting said protein kinase C with an effective amount of a balanoid or a pharmaceutically acceptable salt thereof. Protein kinase C
inhibitors are useful as anti-inflammatory, antitumor, and reperfusion injury agents through their antiproliferative and anti-inflammatory activities in human neutrophils, human keratinocytes, and human tumor cells. The present invention relates to methods of treating animals, specifically mammals, suffering from inflammatory, cardiovascular and/or neoplastic diseases by administering an amount of a balanoid or a pharma-ceutically acceptable salt thereof to the animal. Humantherapeutics are preferred.
The methods of the present invention comprise inhibit-ing protein kinAs~ C activity by contacting protein kinase C
with an inhibitory effective amount of a balanoid. Balanoids been discovered to inhibit the activity of protein kinase C.
Exposure of cells in vitro to balanoids results in the inhibition of PKC activity. Inhibition of PKC activity in cells impedes cellular activities associated with several disease conditions. Of particular note is the selectivity exhibited by Balanoids which permits selective inhibition of one or more isoforms (isozymes) of PKC to a greater degree than other isoforms. Such selectivity has long been desired and is indicative of great therapeutic usefulness.
The methods of the present invention are useful in the treatment of diseases which involve cellular growth, regulation and differentiation such as inflammatory, cardiovascular and neoplastic diseases. PKC activity is associated with disease W094l20062 PCT~S94/02283 2157~112 - 8 -conditions such as cancer, inflammation and reperfusion injury.
Accordingly, the present invention relates to methods of treating a mammal suffering from cancer, inflammation such as the type associated with arthritis, reperfusion injury or other PKC-linked conditions. The methods comprise administering to the mammal an effective amount of a balanoid or a phar-maceutically acceptable salt thereof which inhibits PKC
activity connected with disease.
PKC phosphorylates certain molecules, referred to herein as phosphorylation acceptor molecules. In order to identify compounds that inhibit PKC activity, an appropriate assay is performed. An exemplary and convenient assay is one in which radio labelled ATP is combined with a phosphorylation acceptor molecule in the presence of PKC and a balanoidal PKC
inhibitor-candidate compound (hereinafter referred to as a "test compound"). Various amounts of test compound are used to determine the level of inhibitory activity that a particular test compound possesses. As a control, radio labelled ATP, phosphorylation acceptor molecule and PKC are combined without test compound. Assay conditions such as pH, salt and cofactor conditions are preferably maintained to be similar to physiological levels in order to duplicate in vivo conditions.
In the assay, if PKC is active, the phosphorylation receptor molecule will be phosphorylated, gaining a radiolabelled phosphorus atom. Thus, the inhibitory activity of the test compound can be determined by incubating PKC, 3ZP-ATP, phosphorylation receptor molecule and test compound and then measuring the level of phosphorylation activity by measuring the level of radioactive phosphorus present in the phosphorylation receptor molecule.
A convenient way to determine the selectivity of PKC
inhibitory activity, test compounds are investigated for cAMP
dependent protein kinase (PKA) inhibitory activity. As in the PKC assay, the level of inhibitory activity is determined by measuring the level of phosphorylation of a phosphorylation acceptor molecule incubated with radiolabelled ATP and PKA.
W094/20062 2 1 3 7 4 1 ~ PCT~S94/02283 _ g _ Preferred PKC inhibitors are selective inhibitors and do not effect the activity of PKA.
In order to investigate the effect that balanoid PKC
inhibitors of the present invention have on cell growth and activity, assays are performed such as to determine human tumor cell growth inhibition, human keratinocyte inhibition and - neutrophil superoxide anion release. Briefly, a human tumor cell growth inhibition assay measures the growth of tumor cells in the presence PKC inhibitors by measuring the incorporation of radiolabelled amino acid in cells. The human keratinocyte inhibition assay measures the proliferation of human epidermal keratinocytes in the same manner as tumor cell growth is measured. Hyperproliferation of keratinocytes is symptomatic of many disease conditions associated with inflammation. The neutrophil superoxide anion release assay measures a PKC
inhibitors ability to block the PMA-induced effects on cells.
The ability of the PKC inhibitors to affect superoxide release by PMA stimulated neutrophils is determined by measuring cytochrome C reduction. Cytochrome C is measured by measuring optical density.
In vivo studies to determine the anti-inflammatory activity of a test substance are conducted using the phorbol 12-myristate 13-acetate (PMA) induced mouse ear edema model which is a mouse model of acute inflammation. Using this model, the efficacy of various test compounds as anti-inflammatory agents are determined.
The compounds of the present invention are referred to herein as balanoids. Novel compounds according to the present invention can be expressed by the formula:
G F E
X K
D ~/ B~B2Z
n(~ )) m A
W094l20062 PCT~S94/02283 2 ~ o -wherein:
A is: CH2, NR , S, SO2 or O;
Bl is: NR , O or CH2;
B2 is: CO, CS, or SO2;
Z is: R , aryl, heteroaryl, substituted aryl or substituted heteroaryl;
D is: NR , O or CH2;
E is: R , aryl, heteroaryl, substituted aryl or substituted heteroaryl;
F is: CO, CS, CH(OR), CH2, O, S or NR ;
G is: R7, aryl, heteroaryl, substituted aryl, substituted heteroaryl or substituted cycloalkyl;
R is: hydrogen or lower alkyl;
X is: CO, CS, CH2, CNR or CCR R ;
R , R , R , R , R , R , R , R and R are, independently, hydrogen, lower alkyl, aryl or JR1l;
R5 is: lower alkyl or aryl;
J is: CO, C=NRl2, SO2 or P(O)O alkyl;
R is: hydrogen, lower alkyl, aryl, alkamino, arylamino, aryloxy or alkoxy;
Rl2 is: straight or branched alkyl, aryl;
m is: 1-4;
n is: 1-4; and m plus n is up to 5;
providing that if m is 3, A is NH, Bl is O, B2 is CO, Z is p-hydroxyphenyl, D is NH, X is CO, and E, F, and G, taken together, are OH HO
HOOC~C~
OH
W094/20062 2 1 ~ 7 ~12 PCT~S94/02283 then n is not 1. Compounds according to the present invention include pharmaceutically acceptable salts of these compounds.
Prodrugs, such as those having carbonates and esters of phenolic groups are also within the scope of the invention.
Lower alkyl means a straight chain, branched or cyclic moiety having from 1 to 6 carbon atoms.
Compounds according to the present invention can have at position A: CH2, NR1, S, SO2 or O. It is preferred that A be NH, CH2, or NRl. When A is NR1, it is preferred that R1 be H or lower alkyl, aryl or JR~ wherein J is CO, C=NH or SOz and R~ is lower alkyl, aryl, alkylamino, arylamino, aryloxy or alkoxy.
Thus, compounds of the present invention can have at position A: CH2, NH, S, SO2, O, NSO2CH3, NSO2phenyl, NCONHphenyl, NCO
phenyl, NCH2phenyl, NCH(CH3)2, NCOCH3, NCOCF3, NSO2-(5-dimethylamino-l-naphthalene, NSO2-1-naphthalene, NSO2-2-naphthalene, NSO2-2-methyl-5-nitrophenyl, NSO2-2-nitrophenyl, NSO2-4-nitrophenyl, NCH=NC(CH3)3, NCONHCH3, Nco(cH2)l4cH3~
NCOOCH2phenyl, NCOOCH2CH(CH3) 2 ~ NCOOC(CH3)3, NCOCH2phenyl, NP=O(OCH2CH3)2, NCH2CH3, NCOOCH3, NSO2CH2phenyl or N(CH2)40H.
Compounds according to the present invention can have at position B1: NR2, O or CH2. It is preferred that B1 be NR or O. B1 is more preferably NH or NCH3.
Compounds according to the present invention-can have at position B2: CO, CS, or SO2. It is preferred that B2 be CO
or CS; B2 is more preferably CO.
Compounds according to the present invention can have at position K: H, or lower alkyl, such as methyl, ethyl or propyl. It is preferred that K be H.
- Compounds according to the present invention can have at position Z: R , aryl, heteroaryl, substituted aryl or substituted heteroaryl. In some embodiments, Z is preferably hydroxy substituted aryl, ether substituted aryl, hydroxy substituted heteroaryl or ether substituted aryl. In some embodiments, Z is pyridine, pyrrole, oxazole, indole, purine, furan, thiophene, pyridazine, pyrimidine, pyrazine, imidazole, W094/20062 PCT~S94102283 21S74~
thiazole, isoxazole, pyrazole, isothiazole, benzene, methyl benzene, dimethyl benzene, trimethyl benzene, tetramethyl benzene, ethyl benzene, tetraethyl benzene, propyl benzene, tetrapropyl benzene, butyl benzene, tetrabutyl benzene, pentyl benzene, tetrapentyl benzene, methoxy benzene, dimethoxy benzene, trimethoxy benzene, tetramethoxy benzene, ethoxy benzene, diethoxy benzene, nitro benzene, dinitro benzene, halo benzene, dihalo benzene, trihalo benzene, tetrahalo benzene, benzene carboxylic acid, benzene dicarboxylic acid, benzamide, benzene diamide, 3,5-dihydroxy benzene, trihydroxy benzene, tetrahydroxy benzene, pentahydroxy benzene, triethoxy benzene, tetraethoxy benzene, propoxy benzene, dipropoxy benzene, tripropoxy benzene, tetra propoxy benzene, aniline, diamino benzene, methoxy pyridine, dimethoxy pyridine, hydroxy pyridine, dihydroxy pyridine, ethoxy pyrrole, dihydroxy pyrrole, dimethoxy indole, hydroxy purine, dimethoxy furan, hydroxy thiophene, methoxy pyridazine, dimethoxy pyridazine, hydroxy pyrimidine, diamido pyrimidine, amido pyrazine, cyanobenzene, butyloxybenzene, hydroxyindole, diethoxy pyrazine, phenyl, quinoline, methoxy quinoline, dimethoxy quinoline, trimethoxy quinoline, hydroxy quinoline, dihydroxy quinoline, ethoxy quinoline, amino quinoline, diamido quinoline, trihalo quinoline, quinoline carboxylic acid, quinazoline, methoxy quinazoline, dimethoxy quinazoline, trimethoxy quinazoline, hydroxy quinazoline, trihydroxy quinazoline, tetraethoxy quinazoline, diamino quinazoline, triamido quinazoline, tetrahalo quinazoline, quinazoline dicarboxylic acid. Z is more preferably p-hydroxy phenyl, p-benzyloxy phenyl, p-benzoate phenyl, p-carboxy phenyl, 4-(2-hydroxyphenylcarbonyl)-3,5-dihydroxyphenyl,p-aminophenyl,4-fluoro phenyl, 4-benzyloxy phenyl, p-methyl phenyl, p-benzyloxycarbonyl phenyl, p-nitrophenyl, 5-benzyloxy-2-indole, 5-hydroxy-2-indole, 3,4-dihydroxy phenyl, 2-benzyloxy phenyl, 2-hydroxyphenyl, phenyl, p-NHSO2CH3phenyl, p-methoxymethyleneoxy phenyl, p-acetoxy phenyl. It is more preferred that Z be substituted phenyl. It is most preferred that Z be p-hydroxy phenyl, p-halophenyl or 5-hydroxy indole.
W O 94/20062 2 ~ 12 PCTrJS94/02283 Compounds according to the present invention can have at position D: NR , O or CH2. It is preferred that D be NR , or o. It is more preferred that D be O or NH.
Compounds according to the present invention can have at position E: R4, aryl, heteroaryl, substituted aryl or substituted heteroaryl. In some embodiments, E is preferably hydroxy substituted aryl, ether substituted aryl, hydroxy substituted heteroaryl or ether substituted aryl. In other embodiments, E may be pyridine, pyrrole, oxazole, indole, purine, furan, thiophene, pyridazine, pyrimidine, pyrazine, imidazole, thiazole, isoxazole, pyrazole, isothiazole, benzene, methyl benzene, dimethyl benzene, trimethyl benzene, tetramethyl benzene, ethyl benzene, tetraethyl benzene, propyl benzene, tetrapropyl benzene, butyl benzene, tetrabutyl benzene, pentyl benzene, tetrapentyl benzene, methoxy benzene, dimethoxy benzene, trimethoxy benzene, tetramethoxy benzene, ethoxy benzene, diethoxy benzene, nitro benzene, dinitro benzene, halo benzene, dihalo benzene, trihalo benzene, tetrahalo benzene, benzene carboxylic acid, benzene dicarboxylic acid, benzamide, benzene diamide, 3,5-dihydroxy benzene, trihydroxy benzene, tetrahydroxy benzene, pentahydroxy benzene, triethoxy benzene, tetraethoxy benzene, propoxy benzene, dipropoxy benzene, tripropoxy benzene, tetra propoxy benzene, aniline, diamino benzene, methoxy pyridine, dimethoxy pyridine, hydroxy pyridine, dihydroxy pyridine, ethoxy pyrrole, dihydroxy pyrrole, dimethoxy indole, hydroxy purine, dimethoxy furan, hydroxy thiophene, methoxy pyridazine, dimethoxy pyridazine, hydroxy pyrimidine, diamido pyrimidine, amido pyrazine, diethoxy pyrazine, phenyl, quinoline, methoxy quinoline, dimethoxy quinoline, trimethoxy quinoline, hydroxy quinoline, dihydroxy quinoline, ethoxy quinoline, amino quinoline, diamido quinoline, trihalo quinoline, quinoline carboxylic acid, quinazoline, methoxy quinazoline, dimethoxy quinazoline, trimethoxy quinazoline, hydroxy quinazoline, trihydroxy quinazoline, tetraethoxy quinazoline, diamino quinazoline, triamido quinazoline, tetrahalo quinazoline, quinazoline dicarboxylic acid, 2-hydroxy benzene, 3-hydroxy W094/20062 PCT~S94/02283 21S~12 - 14 -benzene, 3-butyloxy benzene, 3-butyloxy-5-hydroxy benzene, 3-hexanoyloxy-5-hydroxy benzene, 3,5-dioctyloxy benzene, 3-octyloxy-5-hydroxy benzene, 3-methoxy-5-hydroxy benzene, 3,5-bis(acetoxy)benzene,3-(methoxycarbonyl)oxy-5-hydroxy benzene,
~ , FIELD OF THE ~Nv~ ON
The present invention relates to the field of treatments for inflammatory, cardiovascular, metabolic, nervous system, viral infectious, neoplastic and other diseases. The invention provides compounds which can inhibit protein kinase C enzymes. More particularly, the present invention relates to novel compounds which are referred to herein as "balanoids".
R~R~.~UND OF THE INVENTION
Protein kinase C (PRC) is a family of calcium- and phospholipid-dependent serine/threonine-specific protein kinases which play an important role in cellular growth control, regulation, and differentiation. Protein kinase C is activated by diacylglycerol (DAG), a neutral lipid, and when activated will transfer the y-phosphate of MgATP to a serine or threonine residue on a substrate protein. The mechanisms of protein kinase C action have been described in U.S. Patent 4,816,450 issued March 28, 1989 to Bell et al., which is incorporated herein by reference.
The activation of protein kinase C has been implicated in several human disease processes, including cancer tumors, inflammation and reperfusion injury. Accordingly, protein kinase C is a target for therapeutic agents useful in treating these conditions.
Cancer is a disease characterized in part by uncontrolled cell growth. Protein kinase C is directly involved in cellular growth control and is believed to be involved in tumor formation. Protein kinase C is fundamental W094/20062 PCT~S94/02283 _ 2 1 5 l ~ 1 2 -- 2 to the processes involved in tumorigenicity, since it is the major high-affinity receptor for endogenous cellular DAGs as well as for several classes of tumor promoters. These tumor promoters also stimulate protein kinase C catalysis. Castagna et al., (1982) J. Biol. Chem. 257:7847, reported direct activation of protein kinase C by tumor promoting phorbol esters.
Protein kinase C is the major, if not exclusive, intracellular receptor of phorbol esters, which are very potent tumor promoters. Phorbol esters and other tumor promoters bind to and activate protein kinase C. Since DAG and phorbol esters interact at the same site, DAGs have been suggested to be the "endogenous phorbol esters", analogous to the opiate receptor where the conservation of a high affinity receptor implied the existence of an endogenous analogue. DAG has been shown to increase the affinity of protein kinase C for Ca 2 and phospholipid and thus activates protein kinase C at cellular levels of these essential cofactors. Extracellular signals including hormones, growth factors, and neurotransmitters are known to stimulate phosphatidylinositol turnover resulting in the generation of IP3 and DAG.
Structures of 40 distinct oncogenes of viral and cellular origin have revealed that oncogenes encode altered forms of normal cellular proteins. Several of the gene products appear related to growth factors or other elements involved in transmembrane signalling. These oncogene products appear to function by altering the level of critical second messengers. Cells transformed with the oncogenes ras, sis, erbB, abl, and src have been shown to contain elevated levels of DAG which is then believed to activate protein kinase C.
Studies on ras transformed cells have shown protein kinase C
activation to be concomitant with elevation of DAG.
Phorbol esters, such as phorbol myristate acetate (PM~A), have complex effects on cells including effects on membrane function, mitogenesis, differentiation, and gene expression. Synthetic DAGs mimic many of the effects of PMA in vitro and inhibitors of protein kinase C have been shown to 215~412 W094/20062 PCT~S94/02283 block PMA-induced effects on cells. Thus, protein kinase C may mediate the actions of certain oncogenes, such as ras, which cause intracellular increases in DAG and concomitant increases in protein kinase C. In addition, activation of protein kinase C leads to the expression of c-myc, c-fos, c-cis, c-fms, nuclear protooncogenes which are important in cell transformation. Overexpression of protein kinase C in NIH 3T3 cells causes altered growth regulation and enhanced tumorige-nicity, and in rat fibroblasts leads to anchorage-independent growth in soft agar. Overexpression of protein kinase C in these cells resulted in tumor formation in animals receiving transplanted cells.
Several studies have shown increased expression of protein kinase C in certain tumor types such as breast and lung carcinomas. Activated protein kinase C has also been detected in human colon carcinomas although increased expression at the gene level was not seen. Topoisomerases are directly modulated by protein kinase C as substrates for the enzyme.
Protein kinase C inhibitors have been reported to potentiate the antitumor activity of chemotherapeutic agents such as cis-platin both in vitro and in vivo (Grunicke, et al.
(1989) Adv. Enzyme Regul. 28:201; and German Offenlegungs-schrift DE 3827974). In addition, it has been suggested that protein kinase C would be a potential target for therapeutic design because of its central role in cell growth (Tritton, T.R. and J.A. Hickman, (1990) Cancer Cells 2:5-102). German Offenlegungsschrift DE 3827974 Al discloses therapeutic preparations comprising a protein kinase C inhibitor in combination with a lipid, a lipid analogue, a cytostatic agent or phospholipase inhibitor which are useful for cancer therapy.
Inflammation and reperfusion injury, particularly pertaining to cardiac injury, are common conditions for which there exists no definitive treatment despite extensive research. Appropriate treatments for these conditions are needed.
Protein kinase C inhibitors have been demonstrated to block platelet aggregation and release of neutrophil activating W094/20062 PCT~S94/02283 ~15~12 - 4 ~
agents such as platelet activating factor (PAF) (Schachtele, et al. (1988) Biochem. Biophy. Res. Commun. 151:542; Hannun, et al. (1987) J. Biol. Chem. 262:13620; Yamada, et al. (1988) Biochem. Pharmacol. 37:1161). Protein kinase C inhibitors have also been shown to inhibit neutrophil activation, and chemotactic migration (McIntyre, et al. (1987) J. Biol Chem.
262:15730; Lambreth, et al. (1988) J. Biol. Chem. 263:3818;
Pittet, et al. (1987) J. Biol. Chem. 262:10072; and Gaudry, et al. (1988) Immunology 63:715), as well as neutrophil degranulation and release of proteolytic enzymes and reactive oxygen intermediates (Wilson, et al. (1986) J. Biol. Chem.
261:12616; Fujita et al. (1986) Biochem. Pharmacol. 35:4555;
Berkow, et al. (1987) J. Leukoc. Biol. 41: 441; Salzer, et al.
(1987) Biochem. Biophys. Res. Commun. 148:747; Kramer, et al.
(1989) J. Biol. Chem. 262:5876; and Dewald, et al. (1989) Biochem. J. 26~:879).
Thus, inhibitors of protein kinase C have the capability of blocking all three of the most significant mechanisms of pathogenesis associated with myocardial reperfusion injury. Protein kinase C is, accordingly, a drug target for therapeutic agents. Additionally, the inhibitory effect of protein kinAse C inhibitors on keratinocytes, and on the oxidative burst in neutrophils, provides an anti-inflammatory effect.
Groth, T., et al., Proc. Adabori Conf: 3rd Ger.-Jap.
Symp. Pept. Chem., E. Wuensch, ed., 91 (1989) disclose ophiocordin, an antibiotic with antifungal activity having certain structural similarities to certain compounds of the invention.
~UNMARY OF ~HE l~v~.,lON
The present invention relates to a novel class of compounds referred to herein as balanoids. Compounds according to the present invention have the following formula:
2157~12 W O 94/20062 PCTrUS94102283 G F E
1' D l~ B B Z
~ ~ 1 2 n(~ ~)m A
wherein:
A is: CH2, NR, S, S 2 or O;
Bl is : NR, O or CH2;
B2 is: C0, CS, or SO2;
Z is: R4, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
D is: NR3, O or CH2;
E is: R5, aryl, heteroaryl, substituted aryl or substituted heteroaryl:
F is : CO , CS , CH (OR6), CH2 , O , S or NR;
G is: R7, aryl, heteroaryl, substituted aryl, substituted heteroaryl or substituted cycloalkyl;
K is: hydrogen or lower alkyl;
X is: CO, CS, CH2, CNR or CCR R
R , R , R , R , R , R , R , R and Rl are, independently, hydrogen, lower alkyl, aryl or JR ;
R5 is: lower alkyl or aryl;
J is: CO, C=NR, S 2 or P(O)0 alkyl;
Rll is: hydrogen, lower alkyl, aryl, alkamino, arylamino, aryloxy or alkoxy;
Rl2 is: straight or branched alkyl, aryl;
m is: 1-4;
n is: 1-4; and m plus n is up to 5;
providing that if m is 3, A is NH, Bl is 0, B2 is CO, Z is p-hydroxyphenyl, D is NH, X is CO, and E, F, and G, taken together, are W094/20062 PCT~S94/02283 215~12 - 6 -OH HO
HOOC ~$ C
OH
then n is not 1.
Additionally, the present invention relates to pharma-ceutically acceptable salts of the above compounds and to formulations comprising the above compounds in pharmaceutically acceptable carriers. Prodrugs such as carbonates and esters of phenolic functional groups and other species metabolizable into compounds of the invention are also considered to be within the scope of the present invention.
The present invention relates to a method of inhibiting protein kinase C activity which comprises contacting protein kinase C with an inhibitory amount of a balanoid of the invention.
The present invention also relates to methods of treating an animal, preferably a mammal, that is suffering from a PRC-related disease, especially an inflammatory, cardiovascular and/or neoplastic diseases by administering an effective amount of a balanoid to the animal.
DET~TT-~n ~ TPTION OF ~K~KK~ EMBODINENT8 This invention is directed to a family of novel compounds denominated "balanoids". Members of the family have been found to exhibit the ability to inhibit enzymes of the family of enzymes known as protein kinase C enzymes.
Selectivity in inhibitors among the isoforms of protein kinase C (PKC) has been shown for balanoids and it is believed that balanoids will be useful in the treatment of disease linked to PKC enzymes.
Methods for preparing balanoids together with synthetic intermediates, are further objects of the invention as are methods for testing for PKC-linked diseases.
W094/20062 215 ~ PCT~S94102283 The present invention relates to balanoids, and their pharmaceutically acceptable salts and formulations. Compounds according to the present invention have been shown to inhibit protein kinase C. PKC inhibitors are known to be useful in the treatment of cancer, inflammatory and reperfusion injury through their antiproliferative and anti-inflammatory activities in human neutrophils, human keratinocytes, and human tumor cells.
The present invention relates to methods of inhibiting protein kinase C activity which comprises contacting said protein kinase C with an effective amount of a balanoid or a pharmaceutically acceptable salt thereof. Protein kinase C
inhibitors are useful as anti-inflammatory, antitumor, and reperfusion injury agents through their antiproliferative and anti-inflammatory activities in human neutrophils, human keratinocytes, and human tumor cells. The present invention relates to methods of treating animals, specifically mammals, suffering from inflammatory, cardiovascular and/or neoplastic diseases by administering an amount of a balanoid or a pharma-ceutically acceptable salt thereof to the animal. Humantherapeutics are preferred.
The methods of the present invention comprise inhibit-ing protein kinAs~ C activity by contacting protein kinase C
with an inhibitory effective amount of a balanoid. Balanoids been discovered to inhibit the activity of protein kinase C.
Exposure of cells in vitro to balanoids results in the inhibition of PKC activity. Inhibition of PKC activity in cells impedes cellular activities associated with several disease conditions. Of particular note is the selectivity exhibited by Balanoids which permits selective inhibition of one or more isoforms (isozymes) of PKC to a greater degree than other isoforms. Such selectivity has long been desired and is indicative of great therapeutic usefulness.
The methods of the present invention are useful in the treatment of diseases which involve cellular growth, regulation and differentiation such as inflammatory, cardiovascular and neoplastic diseases. PKC activity is associated with disease W094l20062 PCT~S94/02283 2157~112 - 8 -conditions such as cancer, inflammation and reperfusion injury.
Accordingly, the present invention relates to methods of treating a mammal suffering from cancer, inflammation such as the type associated with arthritis, reperfusion injury or other PKC-linked conditions. The methods comprise administering to the mammal an effective amount of a balanoid or a phar-maceutically acceptable salt thereof which inhibits PKC
activity connected with disease.
PKC phosphorylates certain molecules, referred to herein as phosphorylation acceptor molecules. In order to identify compounds that inhibit PKC activity, an appropriate assay is performed. An exemplary and convenient assay is one in which radio labelled ATP is combined with a phosphorylation acceptor molecule in the presence of PKC and a balanoidal PKC
inhibitor-candidate compound (hereinafter referred to as a "test compound"). Various amounts of test compound are used to determine the level of inhibitory activity that a particular test compound possesses. As a control, radio labelled ATP, phosphorylation acceptor molecule and PKC are combined without test compound. Assay conditions such as pH, salt and cofactor conditions are preferably maintained to be similar to physiological levels in order to duplicate in vivo conditions.
In the assay, if PKC is active, the phosphorylation receptor molecule will be phosphorylated, gaining a radiolabelled phosphorus atom. Thus, the inhibitory activity of the test compound can be determined by incubating PKC, 3ZP-ATP, phosphorylation receptor molecule and test compound and then measuring the level of phosphorylation activity by measuring the level of radioactive phosphorus present in the phosphorylation receptor molecule.
A convenient way to determine the selectivity of PKC
inhibitory activity, test compounds are investigated for cAMP
dependent protein kinase (PKA) inhibitory activity. As in the PKC assay, the level of inhibitory activity is determined by measuring the level of phosphorylation of a phosphorylation acceptor molecule incubated with radiolabelled ATP and PKA.
W094/20062 2 1 3 7 4 1 ~ PCT~S94/02283 _ g _ Preferred PKC inhibitors are selective inhibitors and do not effect the activity of PKA.
In order to investigate the effect that balanoid PKC
inhibitors of the present invention have on cell growth and activity, assays are performed such as to determine human tumor cell growth inhibition, human keratinocyte inhibition and - neutrophil superoxide anion release. Briefly, a human tumor cell growth inhibition assay measures the growth of tumor cells in the presence PKC inhibitors by measuring the incorporation of radiolabelled amino acid in cells. The human keratinocyte inhibition assay measures the proliferation of human epidermal keratinocytes in the same manner as tumor cell growth is measured. Hyperproliferation of keratinocytes is symptomatic of many disease conditions associated with inflammation. The neutrophil superoxide anion release assay measures a PKC
inhibitors ability to block the PMA-induced effects on cells.
The ability of the PKC inhibitors to affect superoxide release by PMA stimulated neutrophils is determined by measuring cytochrome C reduction. Cytochrome C is measured by measuring optical density.
In vivo studies to determine the anti-inflammatory activity of a test substance are conducted using the phorbol 12-myristate 13-acetate (PMA) induced mouse ear edema model which is a mouse model of acute inflammation. Using this model, the efficacy of various test compounds as anti-inflammatory agents are determined.
The compounds of the present invention are referred to herein as balanoids. Novel compounds according to the present invention can be expressed by the formula:
G F E
X K
D ~/ B~B2Z
n(~ )) m A
W094l20062 PCT~S94/02283 2 ~ o -wherein:
A is: CH2, NR , S, SO2 or O;
Bl is: NR , O or CH2;
B2 is: CO, CS, or SO2;
Z is: R , aryl, heteroaryl, substituted aryl or substituted heteroaryl;
D is: NR , O or CH2;
E is: R , aryl, heteroaryl, substituted aryl or substituted heteroaryl;
F is: CO, CS, CH(OR), CH2, O, S or NR ;
G is: R7, aryl, heteroaryl, substituted aryl, substituted heteroaryl or substituted cycloalkyl;
R is: hydrogen or lower alkyl;
X is: CO, CS, CH2, CNR or CCR R ;
R , R , R , R , R , R , R , R and R are, independently, hydrogen, lower alkyl, aryl or JR1l;
R5 is: lower alkyl or aryl;
J is: CO, C=NRl2, SO2 or P(O)O alkyl;
R is: hydrogen, lower alkyl, aryl, alkamino, arylamino, aryloxy or alkoxy;
Rl2 is: straight or branched alkyl, aryl;
m is: 1-4;
n is: 1-4; and m plus n is up to 5;
providing that if m is 3, A is NH, Bl is O, B2 is CO, Z is p-hydroxyphenyl, D is NH, X is CO, and E, F, and G, taken together, are OH HO
HOOC~C~
OH
W094/20062 2 1 ~ 7 ~12 PCT~S94/02283 then n is not 1. Compounds according to the present invention include pharmaceutically acceptable salts of these compounds.
Prodrugs, such as those having carbonates and esters of phenolic groups are also within the scope of the invention.
Lower alkyl means a straight chain, branched or cyclic moiety having from 1 to 6 carbon atoms.
Compounds according to the present invention can have at position A: CH2, NR1, S, SO2 or O. It is preferred that A be NH, CH2, or NRl. When A is NR1, it is preferred that R1 be H or lower alkyl, aryl or JR~ wherein J is CO, C=NH or SOz and R~ is lower alkyl, aryl, alkylamino, arylamino, aryloxy or alkoxy.
Thus, compounds of the present invention can have at position A: CH2, NH, S, SO2, O, NSO2CH3, NSO2phenyl, NCONHphenyl, NCO
phenyl, NCH2phenyl, NCH(CH3)2, NCOCH3, NCOCF3, NSO2-(5-dimethylamino-l-naphthalene, NSO2-1-naphthalene, NSO2-2-naphthalene, NSO2-2-methyl-5-nitrophenyl, NSO2-2-nitrophenyl, NSO2-4-nitrophenyl, NCH=NC(CH3)3, NCONHCH3, Nco(cH2)l4cH3~
NCOOCH2phenyl, NCOOCH2CH(CH3) 2 ~ NCOOC(CH3)3, NCOCH2phenyl, NP=O(OCH2CH3)2, NCH2CH3, NCOOCH3, NSO2CH2phenyl or N(CH2)40H.
Compounds according to the present invention can have at position B1: NR2, O or CH2. It is preferred that B1 be NR or O. B1 is more preferably NH or NCH3.
Compounds according to the present invention-can have at position B2: CO, CS, or SO2. It is preferred that B2 be CO
or CS; B2 is more preferably CO.
Compounds according to the present invention can have at position K: H, or lower alkyl, such as methyl, ethyl or propyl. It is preferred that K be H.
- Compounds according to the present invention can have at position Z: R , aryl, heteroaryl, substituted aryl or substituted heteroaryl. In some embodiments, Z is preferably hydroxy substituted aryl, ether substituted aryl, hydroxy substituted heteroaryl or ether substituted aryl. In some embodiments, Z is pyridine, pyrrole, oxazole, indole, purine, furan, thiophene, pyridazine, pyrimidine, pyrazine, imidazole, W094/20062 PCT~S94102283 21S74~
thiazole, isoxazole, pyrazole, isothiazole, benzene, methyl benzene, dimethyl benzene, trimethyl benzene, tetramethyl benzene, ethyl benzene, tetraethyl benzene, propyl benzene, tetrapropyl benzene, butyl benzene, tetrabutyl benzene, pentyl benzene, tetrapentyl benzene, methoxy benzene, dimethoxy benzene, trimethoxy benzene, tetramethoxy benzene, ethoxy benzene, diethoxy benzene, nitro benzene, dinitro benzene, halo benzene, dihalo benzene, trihalo benzene, tetrahalo benzene, benzene carboxylic acid, benzene dicarboxylic acid, benzamide, benzene diamide, 3,5-dihydroxy benzene, trihydroxy benzene, tetrahydroxy benzene, pentahydroxy benzene, triethoxy benzene, tetraethoxy benzene, propoxy benzene, dipropoxy benzene, tripropoxy benzene, tetra propoxy benzene, aniline, diamino benzene, methoxy pyridine, dimethoxy pyridine, hydroxy pyridine, dihydroxy pyridine, ethoxy pyrrole, dihydroxy pyrrole, dimethoxy indole, hydroxy purine, dimethoxy furan, hydroxy thiophene, methoxy pyridazine, dimethoxy pyridazine, hydroxy pyrimidine, diamido pyrimidine, amido pyrazine, cyanobenzene, butyloxybenzene, hydroxyindole, diethoxy pyrazine, phenyl, quinoline, methoxy quinoline, dimethoxy quinoline, trimethoxy quinoline, hydroxy quinoline, dihydroxy quinoline, ethoxy quinoline, amino quinoline, diamido quinoline, trihalo quinoline, quinoline carboxylic acid, quinazoline, methoxy quinazoline, dimethoxy quinazoline, trimethoxy quinazoline, hydroxy quinazoline, trihydroxy quinazoline, tetraethoxy quinazoline, diamino quinazoline, triamido quinazoline, tetrahalo quinazoline, quinazoline dicarboxylic acid. Z is more preferably p-hydroxy phenyl, p-benzyloxy phenyl, p-benzoate phenyl, p-carboxy phenyl, 4-(2-hydroxyphenylcarbonyl)-3,5-dihydroxyphenyl,p-aminophenyl,4-fluoro phenyl, 4-benzyloxy phenyl, p-methyl phenyl, p-benzyloxycarbonyl phenyl, p-nitrophenyl, 5-benzyloxy-2-indole, 5-hydroxy-2-indole, 3,4-dihydroxy phenyl, 2-benzyloxy phenyl, 2-hydroxyphenyl, phenyl, p-NHSO2CH3phenyl, p-methoxymethyleneoxy phenyl, p-acetoxy phenyl. It is more preferred that Z be substituted phenyl. It is most preferred that Z be p-hydroxy phenyl, p-halophenyl or 5-hydroxy indole.
W O 94/20062 2 ~ 12 PCTrJS94/02283 Compounds according to the present invention can have at position D: NR , O or CH2. It is preferred that D be NR , or o. It is more preferred that D be O or NH.
Compounds according to the present invention can have at position E: R4, aryl, heteroaryl, substituted aryl or substituted heteroaryl. In some embodiments, E is preferably hydroxy substituted aryl, ether substituted aryl, hydroxy substituted heteroaryl or ether substituted aryl. In other embodiments, E may be pyridine, pyrrole, oxazole, indole, purine, furan, thiophene, pyridazine, pyrimidine, pyrazine, imidazole, thiazole, isoxazole, pyrazole, isothiazole, benzene, methyl benzene, dimethyl benzene, trimethyl benzene, tetramethyl benzene, ethyl benzene, tetraethyl benzene, propyl benzene, tetrapropyl benzene, butyl benzene, tetrabutyl benzene, pentyl benzene, tetrapentyl benzene, methoxy benzene, dimethoxy benzene, trimethoxy benzene, tetramethoxy benzene, ethoxy benzene, diethoxy benzene, nitro benzene, dinitro benzene, halo benzene, dihalo benzene, trihalo benzene, tetrahalo benzene, benzene carboxylic acid, benzene dicarboxylic acid, benzamide, benzene diamide, 3,5-dihydroxy benzene, trihydroxy benzene, tetrahydroxy benzene, pentahydroxy benzene, triethoxy benzene, tetraethoxy benzene, propoxy benzene, dipropoxy benzene, tripropoxy benzene, tetra propoxy benzene, aniline, diamino benzene, methoxy pyridine, dimethoxy pyridine, hydroxy pyridine, dihydroxy pyridine, ethoxy pyrrole, dihydroxy pyrrole, dimethoxy indole, hydroxy purine, dimethoxy furan, hydroxy thiophene, methoxy pyridazine, dimethoxy pyridazine, hydroxy pyrimidine, diamido pyrimidine, amido pyrazine, diethoxy pyrazine, phenyl, quinoline, methoxy quinoline, dimethoxy quinoline, trimethoxy quinoline, hydroxy quinoline, dihydroxy quinoline, ethoxy quinoline, amino quinoline, diamido quinoline, trihalo quinoline, quinoline carboxylic acid, quinazoline, methoxy quinazoline, dimethoxy quinazoline, trimethoxy quinazoline, hydroxy quinazoline, trihydroxy quinazoline, tetraethoxy quinazoline, diamino quinazoline, triamido quinazoline, tetrahalo quinazoline, quinazoline dicarboxylic acid, 2-hydroxy benzene, 3-hydroxy W094/20062 PCT~S94/02283 21S~12 - 14 -benzene, 3-butyloxy benzene, 3-butyloxy-5-hydroxy benzene, 3-hexanoyloxy-5-hydroxy benzene, 3,5-dioctyloxy benzene, 3-octyloxy-5-hydroxy benzene, 3-methoxy-5-hydroxy benzene, 3,5-bis(acetoxy)benzene,3-(methoxycarbonyl)oxy-5-hydroxy benzene,
3,5-dihydroxy phenyl, 3-ethoxy-5-hydroxy phenyl, 3,5-dibenzyloxy phenyl, 3,5-dimethoxy phenyl, 3-hydroxy-5-benzoate phenyl, phenyl, 3,5-dimethoxymethyleneoxy phenyl, 3-methoxycarbonyloxy phenyl, 3-acetoxy-5-hydroxy phenyl. It is more preferred that E be 3,5-hydroxy benzene or 3-acyloxy-5-hydroxy benzene. It is most preferred that E be 3,5-hydroxy benzene.
Compounds according to the present invention can have at position F: CO, CS, CH(OR6), CH2, O, S or NR6. It is preferred that F be CO or CH2. It is most preferred that F be CO.
Compounds according to the present invention can have at position G: R~, aryl, heteroaryl, substituted aryl, substituted heteroaryl or substituted cycloalkyl. In some embodiments, G is preferably hydroxy substituted aryl, carboxy substituted aryl, hydroxy substituted heteroaryl or carboxy substituted heteroaryl. In other embodiments, G may be pyridine, pyrrole, oxazole, indole, purine, furan, thiophene, pyridazine, pyrimidine, pyrazine, imidazole, thiazole, isoxazole, pyrazole, isothiazole, benzene, methyl benzene, dimethyl benzene, trimethyl benzene, tetramethyl benzene, ethyl benzene, tetraethyl benzene, propyl benzene, tetrapropyl benzene, butyl benzene, tetrabutyl benzene, pentyl benzene, tetrapentyl benzene, methoxy benzene, dimethoxy benzene, trimethoxy benzene, tetramethoxy benzene, ethoxy benzene, diethoxy benzene, nitro benzene, dinitro benzene, halo benzene, dihalo benzene, trihalo benzene, tetrahalo benzene, benzene carboxylic acid, benzene dicarboxylic acid, benzamide, benzene diamide, 3,5-dihydroxy benzene, trihydroxy benzene, tetrahydroxy benzene, pentahydroxy benzene, triethoxy benzene, tetraethoxy benzene, propoxy benzene, dipropoxy benzene, tripropoxy benzene, tetra propoxy benzene, aniline, diamino benzene, methoxy pyridine, dimethoxy pyridine, hydroxy W094t20062 21~ ~ ~1~ PCTtUS94/02283 pyridine, dihydroxy pyridine, ethoxy pyrrole, dihydroxy pyrrole, dimethoxy indole, hydroxy purine, dimethoxy furan, hydroxy thiophene, methoxy pyridazine, dimethoxy pyridazine, hydroxy pyrimidine, diamido pyrimidine, amido pyrazine, 5 diethoxy pyrazine, phenyl, quinoline, methoxy quinoline, dimethoxy quinoline, trimethoxy quinoline, hydroxy quinoline, dihydroxy quinoline, ethoxy quinoline, amino quinoline, diamido quinoline, trihalo quinoline, quinoline carboxylic acid, quinazoline, methoxy quinazoline, dimethoxy quinazoline, trimethoxy quinazoline, hydroxy quinazoline, trihydroxy quinazoline, tetraethoxy quinazoline, diamino quinazoline, triamido quinazoline, tetrahalo quinazoline, quinazoline dicarboxylic acid, 2-cyano-6-hydroxy benzene,, 2-hydroxy-5,6,7,8-tetrahydro-naphthalene,2-acetoxy-6-carboxybenzene,2-hydroxy-6-tetrazolylbenzene,2-hydroxy-naphthalene,2-hydroxy-6-(methoxycarbonyl) benzene, 2-carboxy-3-pyridinyl, 2-(ethoxycarbonyl)-6-hydroxy benzene, 2,3-dihydroxy benzene, 2-carboxy cyclohexane, 2,6-dihalo benzene, 2-acetoxy-6-(ethoxycarbonyl) benzene. In some embodiments, G is preferably 2-carboxy-6-hydroxy phenyl, 2-ethoxycarbonyl-6-hydroxy phenyl, 2-hydroxy phenyl, 2-benzyloxycarbonyl phenyl, 2-hydroxy naphthyl, 2,3,5,6,-tetramethyl phenyl, 2,6-dihydroxy phenyl, 2,6-dimethoxy phenyl, 2-carboxy cyclohexane, 2-hydroxy cyclohexane, 2-hydroxy-1-naphthyl, 2,6-dichloro phenyl, 2-methoxy-6-hydroxy phenyl, 2-carboxy-3-pyridine, 3-carboxy-2-pyridine, phenyl, 3,4-dihydroxy phenyl, 2-methoxycarbonyl-6-hydroxy phenyl, 2-butoxycarbonyl-6-hydroxy phenyl, 2-(2-methylpropyloxycarbonyl)-6-hydroxyphenyl, 2-nitrilo-6-hydroxy phenyl, 2-carboxy phenyl, 2-(4-acetoxybenzyloxycarbonyl)-6-hydroxy phenyl, 2-benzyloxycarbonyl-6-benzyloxy phenyl, 2,6-dibenzyloxy phenyl, 2-benzyloxycarbonyl cyclohexane, 1-benzyloxy-2-naphthyl, 2-methoxy-6-benzyloxy phenyl, 2-benzyloxycarbonyl-3-pyridinyl, 3-benzyloxycarbonyl-2-pyridinyl, 2-benzyloxyphenyl, 2-nitrilo-6-benzyloxy phenyl, 3,4-dibenzyloxyphenyl, 2-benzyloxy-1-naphthyl, 6-benzyloxy-2-tetrazolylphenyl, 6-hydroxy-2-tetrazolylphenyl, 2-methyltetrazolylphenyl,3-methyltetrazolylphenyl,2-hydroxy-1-W O 94/20062 PCTrUS94/02283 2~ 2 - 16 -(5,6,7,8-tetrahydro) naphthyl,3-benzyloxycarbonyl-4-benzyloxy phenyl, 3-carboxy-4-hydroxy phenyl, 2-methoxymethyleneoxy phenyl, 2-ethoxycarbonyl-6-benzyloxy phenyl, 2-benzyloxy carbonyl-l-naphthyl, 2-carboxy-1-naphthyl, 2-benzyloxy-6-methyl phenyl, 2-methyl-6-hydroxy phenyl, 2-acetoxy-6-ethoxycarbonyl phenyl, 2-cyclohexylmethoxycarbonyl-6-hydroxy phenyl, 2-carboxy-6-benzyloxy phenyl, 2-methoxycarbonyl-6-benzyloxy phenyl, 2-hexanoyloxy-6-carboxy phenyl. It is more preferred that G be 2-carboxy-6-hydroxyphenyl, 2-hydroxy-6-(tetrazol-2-y)phenyl, 2,6-dihydroxy phenyl, 2-hydroxy-1-naphthyl, 2-methoxycarbonyl-6hydroxyphenyl, 2-cyano-6-hydroxy phenyl, and 2-hydroxy-6-(trifluoromethylsulfonamino)phenyl. It is most preferred that G be 2-carboxy-6-hydroxy benzene and its ester or acyl derivatives as well as 2-R-6-hydroxyphenyl where R is carboxylic acid surrogate such as tetrazole or N-sulfonylcarboxamide.
Compounds according to the present invention can have at position X: CO, CS, CNR3 or CCR9R10. It is preferred that X
be C0 or CH2.
Compounds according to the present invention can have 2 3 4 6 R7 R3 R9 and Rl, independently: Y
lower alkyl, aryl or JRl1 wherein J is CO, CN or S02 and R11 is lower alkyl, aryl, alkylamino, arylamino, aryloxy or alkoxy.
Compounds according to the present invention can have at position R5 lower alkyl or aryl.
In compounds according to the present invention, m is 1-4. It is preferred that m be 1-2, preferably 1.
In compounds according to the present invention, n is 1-4. It is preferred that n be 1-3.
In compounds according to the present invention, n plus m is less than or equal to 5. It is preferred that n plus m is less than or equal to 4.
It will be appreciated that atoms within the moieties defined by n and m may have substituents. Such substituents may preferably include hydrocarbyl groups such as the lower alkyl groups, methyl, ethyl and propyl together with larger aliphatic and aromatic functions.
WOg4/20062 2 i ~ 7 `1~ 2 PCT~S94/02283 It will also be appreciated that m and n may contain other functional species such as carbonyl, thiocarbonyl, hydroxy, amino, halo and others. Preferred species are carbonyl and thiocarbonyl.
Preferred compounds contain aryl groups in positions Z, E and G. In preferred compounds, Z, E and G are hydroxy substituted aryl, ether substituted aryl, hydroxy substituted heteroaryl, or ether substituted aryl. E and G are preferably substituted aryls or substituted heteroaryls whereby the substitutions are located such that the two aryl rings are conformationally "crowded" out of plane with each other.
Additionally, A is preferably NR or CHz.
Pharmaceutically acceptable salts of these compounds may be used in accordance with the present invention. One having ordinary skill in the art could readily appreciate what salts would be appropriate. Pharmaceutically acceptable salts include, but are not limited to sodium, trialkyl ammonium, potassium, calcium, zinc, lithium, magnesium, aluminum, diethanolamine, ethylenediamine, meglumine and acetate.
Preferred salts are sodium and potassium.
In certain preferred compounds of the present inven-tion A is CH2, NR , S, or O; Bl is NR , O, or CH2; B2 is CO or CS; Z is R4, aryl, heteroaryl, substituted aryl or substituted heteroaryl or D is NR3, O or CH2; E is R5, aryl, heteroaryl, substituted aryl or substituted heteroaryl; F is CO or CS; G is R7, aryl, heteroaryl, substituted aryl or substituted heteroaryl; X is CO or CS; R, R, R, R, R, R, R, R and R
are, independently: hydrogen, lower alkyl or aryl; R5 is lower alkyl or aryl; m is 1-4; and n is 1-4; where n plus m is less than or equal to 5.
In other preferred compounds of the present invention A is CH2, NRl, S, or O; Bl is NR2 or O; B2 is CO or CS; Z is hydroxy substituted aryl, ether substituted aryl, hydroxy substituted heteroaryl, halo substituted aryl; D is NR3 or 0;
E is hydroxy substituted aryl, ether substituted aryl, hydroxy substituted heteroaryl, acyloxy substituted aryl; F is CO or CS; G is hydroxy substituted aryl, ether substituted aryl, W094/20062 PCT~S94/02283 2 1 ~ 7 ~1~ 18 -hydroxy substituted heteroaryl, carboxy substituted aryl; X is CO or CS; Rl, R2 and R3 independently: hydrogen, lower alkyl or aryl; m is 1-2; and n is 1-3; where n plus m is less than or equal to 4.
In still other preferred compounds of the present invention A is NH , CH2 or NR; Bl is NR or O; B2 is CO or CS;
Z is hydroxyphenyl or halophenyl; D is NR3, O or CH2; E is 3,5-hydroxy benzene or 3,5-alkoxy benzene; F is CO or CH2; G is 2-carboxy-benzene, 2-hydroxy benzene, 2,6-dihydroxy benzene, 2-methoxy benzene, 2,6-dimethoxy benzene, or 6-hydroxy benzene-2-carboxylic acid; X is CO; Rl, R2, or R3 are, independently hydrogen, lower alkyl or aryl; m is l; and n is 3.
In certain preferred compounds of the present inven-tion A is NH or CH2; Bl is NH; B2 is CO; Z is p-hydroxyphenyl;
15 D is O; E is 3,5-hydroxy benzene; F is CO; G is 2-carboxy-6-hydroxyphenyl, 2-hydroxy-6-(tetrazol-2-y)phenyl, 2,6-dihydroxy phenyl, 2-hydroxy-1-naphthyl, 2-methoxycarbonyl-6hydroxyphenyl, 2-cyano-6-hydroxy phenyl, and 2-hydroxy-6-(trifluoromethylsulfonamino)phenyl; X is CO; m is l; and n is 3.
As will be appreciated, it is generally the case that one stereoisomer is more biologically active than its enantiomer. It is envisioned that preferred stereoisomerism will be determined for active species and that such preferred compounds will be selected for therapeutic and other uses.
Compounds of the present invention may be synthesized from readily available starting materials by st~n~rd ~er-~n;ques such as by following the basic synthesis set out below. One having ordinary skill in the art may employ other well known synthetic schemes to produce compounds according to the present invention.
Prodrugs such as carbonates and carboxy esters of phenolic OH and NH groups can be prepared by the derivatization of OH and NH groups with acylating agents, such as methyl chloroformate, ethyl chloroformate, isobutyryl chloride, methoxypropionyl chloride, methyl chlorosuccinate, ethyl chlorosuccinate and benzoyl chloride, for example.
21~74~ 2 W094l20062 PCT/US94/02283 Additionally, prodrugs of compounds which contain a carboxylic acid can be prepared by derivitazation with alkylating agents, such as methyl iodide or acetoxymethyl chloride.
Reaction Scheme I provides syntheses for producing compounds according to the present invention including the use of a cyclic carbonyl or a heterocyclic carbonyl such as the seven membered lactam shown as a starting material. For example, a lactam can be benzylated with base in tetrahydrofuran to protect the nitrogen functionality. It is then reacted with base and phenylselenyl chloride followed by sodium periodate to yield the unsaturated lactam. Oxidation with osmium tetroxide followed by benzoylation yields the hydroxy benzoate shown. Further reaction with trifluoromethanesulfonic anhydride and sodium azide provides the anti azido ester which can be reduced with, for example, lithium aluminum hydride. Reaction with a carboxylic acid substituted with a Z functionality yields the amide. Further reaction with a GFE carboxylic function followed by deprotection provides the family of ester/amides.
2157~112 -20-O
_~z = ~ Z ti `C
T ~ T
O t_ ~Z= e_~
.' T ~,, o T
m . .
~ m ~:
Q IL Q I
Z Jl-- ~ O
C~ 11 -- ' --C
W094/20062 ~i 7 11~ PCT~S94/02283 Scheme II provides a synthesis scheme for producing compounds according to the present invention including the use of an enol ether lactam such as the azepinone shown as a starting material. For example, an enolether lactam can be benzylated with base in tetrahydrofuran to protect the nitrogen function. It is then hydrolyzed with acid and reacted with sodium nitrite in acetic acid to form the oxime. Catalytic hydrogenation in the presence of acetic anhydride gives the acetamide, which can be reduced with, for example, sodium borohydride and hydrolyzed to the syn (or anti) aminohydroxy lactam. Reduction with, for example, lithium aluminum hydride and reaction with a carboxylic acid substituted with a Z
functionality yields the amide, which can be further reacted with GFE carboxylic function and deprotected to provide the family of ester/amides.
WO 94noo62 rCT/US94/02283 2 1 5 7 ~c~jeme II
OEt OEt ~O
1)KOtBu ~ HCUH20 ~
~ N ~ o 2)PhCH28r Ph N~ ~h Y.Tamuraetal., Chem.Pharm.Bull., ~(3).523-8,529-34(1971) 1 ) NaBH4 ~2)HCUH20/EtOH
NaNO2 ~ NOH H2/RaneyNi ~ NHCOCH3 3) NaOH
ACOH/Ac20 ~ N ~ O AcOHlAc20 ~ O
Ph Ph H ~OH Z-COCI OH
NHz LAH (~ IH2 Et3N/DMAP(~;~NHCO--Z
Ph Ph Z-C-N ~N Ph G-F-E-COCI
Et3NtDMAP lc_E-F-G H2 olC_E-F-G
~ ~NHCO - Z
G-F-E-COOH ( ~ NHCO - Z Pd(OH)2;C H
DCCIDMAP
W094/20062 21~ 2 PCT~S94/02283 Reaction Scheme III provides methods for producing compounds according to the present invention including the use as a starting material of the previously mentioned syn aminohydroxy lactam. Oxidation of the alcohol using, for example, oxalyl chloride, dimethylsulfoxide, and triethylamine (Swen oxidation) provides the Keto intermediate which is treated with HONH2-HCl followed by reduction using, for example, RaNi (Raney nickel) catalyst and hydrogen affords the amino-amide. Reaction with a GFE carboxylic acid function followed by deprotection provides the family of diamides.
215 7 ~ 24 - PCTIUS9~/02283 Scheme m ~OH O
(~NHCO--Z oxalyl chloride ~NHCO Z 1) NH20H-HCI
DMSO, Et3N, -65C NJ 2) RaNi. H2 Ph Ph G-F-E-COCI
~NH2 Rm Et3N/DMAP NHCO-E-F-G
(;;)~NHCO{~ or O~ NHCO--Z
G-F-E-co-N~g N
Ph Ph NHCO-E-F-G
~NHCO Z
Pd(OH)2/C H
W094eO062 ~ 3 5, ~12 PCT1594/02283 Scheme IV provides a synthesis scheme for producing compounds according to the present invention including the use as a starting material of a cyclic olefin as shown.
Epoxidation with peracetic acid followed by reaction with sodium azide affords the anti azido alcohol, which is O-protected and reduced to the aminoalcohol ether. Reaction with a carboxylic acid substituted with a Z functionality yields the amide, which is O-deprotected and reacted with a GFE carboxylic function to provide the family of ester/amides.
WO 94/20062 rCT/US94/02283 2 1 5 ~ 26 -t~ T
=:E O ~
IL -~3 ~
m ~ ~0~
~0 s _ c ~
_ c T
T
Z~
, E m ~
z~
~ o T T
0~ ~
N _ T
~ o U~f~O
z 5 0 C~ ~ Z`'-o , ~ r~l ~0 W094/2006~ 2 1 ~ 2 _ PCT/US94/02283 Scheme V provides a syntheses for preparing compounds according to the present invention including a synthesis scheme for producing 6, 7 and 8 member cyclic and heterocyclic groups including Bl and D with stereo specific attachment, including the use as a starting material of the unsaturated aldehyde.
For example, tin-mediated condensation and subsequent cyclization of the aldehyde with an isothiocyanate affords the oxazolidine thione, which can be reduced with lithium aluminum hydride, ozonolyzed, and further reduced with, for example, sodium borohydride to the diol. Mesylation with methanesulfonyl chloride and base followed by ring closure with benzylamine affords the azepine, which is hydrolyzed with an acid, for example, hydrochloric açid. The resultant anti aminoalcohol is reacted with a carboxylic acid substituted with a Z functionality to give the amide, which is further reacted with a GFE carboxylic function and deprotected to provide the family of ester/amides with predictable stereochemistry at the positions of attachment of B1 and D to the ring system.
WO 94/2006~
21~7 4~ -28 ô
CL
o~ ~ I I
Z
m ~<O~
--~--LIJ_~
~O~cn z J Z
~ .
T I oN
> ,_ Z O ~
O=~ c ~ ~ U e~
0=( ~ 5 O ~<O~
N~ 3~ ~> O
O ~ ,~
Z N m , N
3~ C`' ~.) C~
0~ ~ (~ _ ~ Z_~m O O
0~
W094/20062 2 1 ~ 7 ~ 1 2 The reactions of Scheme VI provide a synthesis scheme for producing 6, 7, and 8 membered cyclic and heterocyclic groups including Bl and D with stereo specific attachment including the use as a starting material of N-carbobenzyloxy asparagine. For example, CBZ-asparagine is reacted with bis(trifluoroacetoxy) iodobenzene to give the mono-protected - diaminoacid, which is differentially protected with di-t-butyl dicarbonate and reduced with, for example, borane/tetrahydrofuran. Oxidation to the aldehyde and condensation with an unsaturated organometallic affords the diprotected diamino alcohol, which gives the terminal tosyloxy compound after hydroboration/oxidation and treatment with toluene sulfonyl chloride. Removal of the butoxycarbonyl is accomplished by acid treatment, for example, formic acid or trifluoroacetic acid, followed by removal of the amine-protecting group, which can be removed with hydrogen.
Selective reaction with a carboxylic acid substituted with a Z
functionality followed by treatment with benzyl chloroformate and base gives the protected amide, which is further reacted with a GFE carboxylic function and deprotected to provide the family of ester/amides with predictable stereochemistry at the positions of attachment of B1 and D to the ring system.
2 1 5 ~ l 1 2 Scheme Vl H~N$~ ~_NHcez DMAP NEI ~~~ ez NHCeZ ~ ~ S wcr~
HN ~aM5CI , HN
80C n=0-2 80C 80C
l) BBN
2) H~02 3) Tsa TsO/~ z) tase ~ NHCeZ ~ Pd~OH) ~ N
Azapine 1.
Int~..c~ with alk...~ti~ synthcsis shown on nc:a shccet SUBSTITUTE SHEET (RULE 263 W094/20062 21~ ~ k ~ ~ PCT/US94102283 Scheme VII provides a syntheses for producing compounds according to the present invention including a synthesis scheme for producing 6, 7, and 8 member cyclic and heterocyclic groups including Bl and D with stereo specific attachment including the use as a starting material of phthalimide alkyl aldehyde. for example, the aldehyde can be reacted with methyl isocyanoacetate in the present of gold ferrocene catalyst to give the oxazolidine, which is hydrolyzed to the diaminohydroxy ester salt with for example, hydrochloric acid. Base mediated cyclization affords the lactam, which can be reduced to an aminohydroxy compound. Reaction with a carboxylic acid substituted with a Z functionality followed by treatment with benzyl chloroformate and base gives the protected amide, with is further reacted with a GFE carboxylic function and deprotected to provide the family of ester/amides with predictable stereo chemistry at the positions of attachment of B1 and D to the ring system.
.'O 94/20062 2~7~12 -32-Scheme Vll O Gold F~ ucc~c O
~N~r~ C ?~'CH,CO2CH3 [~ ~; O
O CHO O ,~
H3CO2C ~N"
n=0-2 1) 6NHCI
2) MeOH, HCI
OH `~
(~ .,NH2 MeOH,K2CO3 yH2-HCI
~/ ~ HCI-H2N ~C02CH3 N ~o OH
LAH
OH 1) ZCOCI OH
~'EL3, D~ N~r z N CBZ
H
Azapinc 1 G-F-E-COCI
~ ~Et3, DMAP
E--F-G
E--F-G O~
o H2, Pd(OH)2 _~ H
~,... ~N~rz lFA.ELOH C8Z
W094/20062 2 ~ ~ 7 ~ ~ 2 rcT~sg~/02283 Scheme VIII A provides a synthesis scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including Bl and D with syn attachment including the se of the protected cyclic ketone as starting material. For example, the ketone is deprotonated and the enolate aminated to afford the butoxycarbonylamino ketone which can be stereo specifically reduced with, for example, sodium borohydride to the syn aminoalcohol. Reactions with a carboxylic acid substituted with a Z functionality yields the amide. Further reaction with a GFE carboxylic function followed by deprotection provides the family of syn ester/amides.
WO 94/20062 PCTtUS9 1tO2283 7 ~ 1 2 34 o I
N m _~ ,O~ZT
Z
m O m ,~ I
._ T
O-~Z~ +' TZ
~ ~0--~Z ~ ~
T _ O
O~Z~ ~
+ + ~
,~ ~ X o X ,.
~ o U U D
-- o~Z~ o o z -W094/20062 21~ PCT~S91/02283 Scheme VIII B provides a synthesis scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including B1 and D with anti attachment including the use as starting material of the syn hydroxyamide from Scheme VIII A. For example, the syn hydroxyamide can be inverted to anti hydroxyamide by treatment with carboxylic acid, such as acetic acid, in the present of triphenylphosphine and diethylazodicarboxylate followed by treatment with sodium methoxide. Reaction with a GFE
carboxylic function followed by deprotection provides the family of anti ester/amides.
WO 9~/20062 PCT/US9~/02283 21~7~12 -36-~
X X IZ ~, U U O ~ ZI Nl C
U
Q
Z
T~ a~ ., O ~ JQ -- o o D D C~
X ~ T
U O
~N
~ IZ '~
o ~Z ~ 11 u _ E
C~
W094120062 21~ ~ ~12 PCT/US94/02283 Scheme VIII C provides methods for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including Bl and D with anti attachment including the use as starting material of the syn hydroxyamide from Scheme VIII A.
For example, the syn hydroxyamide can b~ inverted to anti amino amide by treatment with trifluoromethane sulfonic anhydride and sodium azide followed by reduction with, for example, tin (II) chloride. Reaction with a GFE carboxylic function followed by deprotection provides the family of anti diamides.
WO 94/2006~
21 ~! 112 -38-_ I
r ~
o Z.. ~ZT c~l C ¦
U U U Z"
~ ' _ o t~l N
m .q Z~Z Z
D O o ~ ~ _ N ~ N
_~ a ~J
W094/20062 2 ~ 3 PCT~S91/02283 Scheme VIII D provides a synthesis' scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including Bl and D with syn attachment including the use as starting material of the anti hydroxyamide from Scheme VIII B. For example, the anti hydroxyamide can be inverted to syn amino amide by treatment with trifluoromethanesulfonic anhydride and sodium azide followed by reduction with, for example, tin (II) chloride.
Reaction with a GFE carboxylic function followed by deprotection provides the family of syn diamides.
~'O 94/20062 N
O
+ <, 3 U O X ~ Z
,, ~, U
N
T
a I_~z~ . Z
N
O
~ o ~ E
W094/20062 21~ 7 41~ PCT/US94/02283 Scheme VIII E provides a synthesis scheme for producing compounds according to the present invention including producing 5 and 6 member cyclic and heterocyclic groups including Bl and D with anti attachment including the use of the protected cyclic epoxide as starting material. For example, the epoxide is opened with ammonia to provide the anti aminoalcohol or with azide to provide the azido-alcohol.
Reduction of the latter with, for example, triphenylphosphine provides the amino alcohol. Reaction with a carboxylic acid substituted with a Z functionality yields the anti hydroxyamide. Further reaction with a GFE carboxylic function followed by deprotection provides the family of anti ester/amides.
WO 94/20062 rCT/US91/02283 2 1 ~ 7 4 1 2 -42-D
X m Ll O U --, GZ I U~l / e x Z I
~ Z~
N ~
m o Z--,G n ~ C~L"_ e ~ ~ ~
D <~ --3 z ~
~, o ~ I o m N ~ z e ~J5 D
Z~~\ n +~
~
S O~z~
WO9ql20062 21~ PCT/159~;02283 Scheme VIII F provides a synthesis scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including Bl and D with syn attachment including the use of the anti hydroxyamide from Scheme VIII E as starting material. For example, the anti hydroxyamide can be inverted to syn hydroxyamide by treatment with carboxylic acid, such as acetic acid in the presence of triphenylphosphine and diethyl azodicarboxylate followed by deprotection provides the family of syn ester/amides.
2~^~7~
o I ~, N
~-) XO T
~ U --_~
~~
X
U
N ~
0~ 0 ~
w O D
X tL
u m O
'~QZ E
E
-C~
W094/20062 2 ~ 12 PCT/US9~/02283 Scheme VIII G provides a synthesis scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including B~ and D with syn attachment including the use of the anti hydroxyamide from Scheme VIII E as starting material. For example, the anti hydroxyamide can be inverted to syn amino amide by treatment with trifluoromethanesulfonic anhydride and sodium azide followed by reduction with, for example, tin (II) chloride.
Reaction with a GFE carboxylic function followed by deprotection provides the family of syn diamides.
2 1~ l 4 12 O
+ <~31 -~ZI
U ,, U
O ~ ~ _ tO ~3 o TZ X~Z ~ Z
N ~_ n --I ~ ~ CL
c~ I ~ _ X S
r~ , U
N
E I E E
U, W094/20062 215 ~ ~ ~ 2 PCT/US94/02283 Scheme VIII H provides a synthesis scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including the use of the syn hydroxyamide from Scheme VIII F as starting material. For example, the syn hydroxyamide can be inverted to anti amino amide by treatment with trifluoromethanesulfonic anhydride and sodium azide followed by reduction with, for example, tin (II) chloride. Reaction with a GFE carboxylic function followed by deprotection provides the family of anti diamides.
WO 9~/20062 2 1 ~ ~ 4 1 2 48 ~I tL
Z .~, I
</~
~ x I .. ~
U U X
o ~ U
o o OI
a ~ t~
~ C D
Z'--~ Z
~ O
D _, Q~ C
U
~ a T--~ ~ ~ N z C~
W094/20062 21~ ~ ~1~ PCT/US9S/02283 Scheme IXA provides a synthesis scheme for producing intermediate GFE carboxylic acids including the use of 4-bromobenzoic acids as starting material. For example a 4-bromobenzoic acid can be esterified by treatment with carbonyldiimidazole followed by an alcohol such as tert-butanol. The ester is treated with n-butyl lithium followed by an addition of N, N-dimethyl formamide to give the aldehyde ester, or by addition of carbon dioxide to give the acid ester, which is converted to the acid chloride ester with oxalyl chloride. The ester can also be reacted with n-butyl lithium and a benzaldehyde to give the diphenyl carbinol, which is oxidized with, for example, chromic acid, and deprotected to provide GFE carboxylic acids. The ester can be reacted with n-butyl lithium and benzoyl chloride to give the diphenyl ketone, which is deprotected to also provide GFE carboxylic acids.
21~12 .so.
Scheme rx A L) n-suLiO c~c~2~
Er~CO2H ,, ~,~C02R --2) CO2~CO2R
~ (COC~).
CIOC~CO2R
X 1) ~ ~CO R
O O
~COzR ~CO2H
Er~ 1) B .I; ~CO2R
o ~CO2H
SUBS~lTUTE SHEET (RULE 2~
2~7412 WO9~/20062 PCT/US91l02283 Scheme IXB provides a synthesis scheme for producing intermediate GFE carboxylic acids including the use of either
Compounds according to the present invention can have at position F: CO, CS, CH(OR6), CH2, O, S or NR6. It is preferred that F be CO or CH2. It is most preferred that F be CO.
Compounds according to the present invention can have at position G: R~, aryl, heteroaryl, substituted aryl, substituted heteroaryl or substituted cycloalkyl. In some embodiments, G is preferably hydroxy substituted aryl, carboxy substituted aryl, hydroxy substituted heteroaryl or carboxy substituted heteroaryl. In other embodiments, G may be pyridine, pyrrole, oxazole, indole, purine, furan, thiophene, pyridazine, pyrimidine, pyrazine, imidazole, thiazole, isoxazole, pyrazole, isothiazole, benzene, methyl benzene, dimethyl benzene, trimethyl benzene, tetramethyl benzene, ethyl benzene, tetraethyl benzene, propyl benzene, tetrapropyl benzene, butyl benzene, tetrabutyl benzene, pentyl benzene, tetrapentyl benzene, methoxy benzene, dimethoxy benzene, trimethoxy benzene, tetramethoxy benzene, ethoxy benzene, diethoxy benzene, nitro benzene, dinitro benzene, halo benzene, dihalo benzene, trihalo benzene, tetrahalo benzene, benzene carboxylic acid, benzene dicarboxylic acid, benzamide, benzene diamide, 3,5-dihydroxy benzene, trihydroxy benzene, tetrahydroxy benzene, pentahydroxy benzene, triethoxy benzene, tetraethoxy benzene, propoxy benzene, dipropoxy benzene, tripropoxy benzene, tetra propoxy benzene, aniline, diamino benzene, methoxy pyridine, dimethoxy pyridine, hydroxy W094t20062 21~ ~ ~1~ PCTtUS94/02283 pyridine, dihydroxy pyridine, ethoxy pyrrole, dihydroxy pyrrole, dimethoxy indole, hydroxy purine, dimethoxy furan, hydroxy thiophene, methoxy pyridazine, dimethoxy pyridazine, hydroxy pyrimidine, diamido pyrimidine, amido pyrazine, 5 diethoxy pyrazine, phenyl, quinoline, methoxy quinoline, dimethoxy quinoline, trimethoxy quinoline, hydroxy quinoline, dihydroxy quinoline, ethoxy quinoline, amino quinoline, diamido quinoline, trihalo quinoline, quinoline carboxylic acid, quinazoline, methoxy quinazoline, dimethoxy quinazoline, trimethoxy quinazoline, hydroxy quinazoline, trihydroxy quinazoline, tetraethoxy quinazoline, diamino quinazoline, triamido quinazoline, tetrahalo quinazoline, quinazoline dicarboxylic acid, 2-cyano-6-hydroxy benzene,, 2-hydroxy-5,6,7,8-tetrahydro-naphthalene,2-acetoxy-6-carboxybenzene,2-hydroxy-6-tetrazolylbenzene,2-hydroxy-naphthalene,2-hydroxy-6-(methoxycarbonyl) benzene, 2-carboxy-3-pyridinyl, 2-(ethoxycarbonyl)-6-hydroxy benzene, 2,3-dihydroxy benzene, 2-carboxy cyclohexane, 2,6-dihalo benzene, 2-acetoxy-6-(ethoxycarbonyl) benzene. In some embodiments, G is preferably 2-carboxy-6-hydroxy phenyl, 2-ethoxycarbonyl-6-hydroxy phenyl, 2-hydroxy phenyl, 2-benzyloxycarbonyl phenyl, 2-hydroxy naphthyl, 2,3,5,6,-tetramethyl phenyl, 2,6-dihydroxy phenyl, 2,6-dimethoxy phenyl, 2-carboxy cyclohexane, 2-hydroxy cyclohexane, 2-hydroxy-1-naphthyl, 2,6-dichloro phenyl, 2-methoxy-6-hydroxy phenyl, 2-carboxy-3-pyridine, 3-carboxy-2-pyridine, phenyl, 3,4-dihydroxy phenyl, 2-methoxycarbonyl-6-hydroxy phenyl, 2-butoxycarbonyl-6-hydroxy phenyl, 2-(2-methylpropyloxycarbonyl)-6-hydroxyphenyl, 2-nitrilo-6-hydroxy phenyl, 2-carboxy phenyl, 2-(4-acetoxybenzyloxycarbonyl)-6-hydroxy phenyl, 2-benzyloxycarbonyl-6-benzyloxy phenyl, 2,6-dibenzyloxy phenyl, 2-benzyloxycarbonyl cyclohexane, 1-benzyloxy-2-naphthyl, 2-methoxy-6-benzyloxy phenyl, 2-benzyloxycarbonyl-3-pyridinyl, 3-benzyloxycarbonyl-2-pyridinyl, 2-benzyloxyphenyl, 2-nitrilo-6-benzyloxy phenyl, 3,4-dibenzyloxyphenyl, 2-benzyloxy-1-naphthyl, 6-benzyloxy-2-tetrazolylphenyl, 6-hydroxy-2-tetrazolylphenyl, 2-methyltetrazolylphenyl,3-methyltetrazolylphenyl,2-hydroxy-1-W O 94/20062 PCTrUS94/02283 2~ 2 - 16 -(5,6,7,8-tetrahydro) naphthyl,3-benzyloxycarbonyl-4-benzyloxy phenyl, 3-carboxy-4-hydroxy phenyl, 2-methoxymethyleneoxy phenyl, 2-ethoxycarbonyl-6-benzyloxy phenyl, 2-benzyloxy carbonyl-l-naphthyl, 2-carboxy-1-naphthyl, 2-benzyloxy-6-methyl phenyl, 2-methyl-6-hydroxy phenyl, 2-acetoxy-6-ethoxycarbonyl phenyl, 2-cyclohexylmethoxycarbonyl-6-hydroxy phenyl, 2-carboxy-6-benzyloxy phenyl, 2-methoxycarbonyl-6-benzyloxy phenyl, 2-hexanoyloxy-6-carboxy phenyl. It is more preferred that G be 2-carboxy-6-hydroxyphenyl, 2-hydroxy-6-(tetrazol-2-y)phenyl, 2,6-dihydroxy phenyl, 2-hydroxy-1-naphthyl, 2-methoxycarbonyl-6hydroxyphenyl, 2-cyano-6-hydroxy phenyl, and 2-hydroxy-6-(trifluoromethylsulfonamino)phenyl. It is most preferred that G be 2-carboxy-6-hydroxy benzene and its ester or acyl derivatives as well as 2-R-6-hydroxyphenyl where R is carboxylic acid surrogate such as tetrazole or N-sulfonylcarboxamide.
Compounds according to the present invention can have at position X: CO, CS, CNR3 or CCR9R10. It is preferred that X
be C0 or CH2.
Compounds according to the present invention can have 2 3 4 6 R7 R3 R9 and Rl, independently: Y
lower alkyl, aryl or JRl1 wherein J is CO, CN or S02 and R11 is lower alkyl, aryl, alkylamino, arylamino, aryloxy or alkoxy.
Compounds according to the present invention can have at position R5 lower alkyl or aryl.
In compounds according to the present invention, m is 1-4. It is preferred that m be 1-2, preferably 1.
In compounds according to the present invention, n is 1-4. It is preferred that n be 1-3.
In compounds according to the present invention, n plus m is less than or equal to 5. It is preferred that n plus m is less than or equal to 4.
It will be appreciated that atoms within the moieties defined by n and m may have substituents. Such substituents may preferably include hydrocarbyl groups such as the lower alkyl groups, methyl, ethyl and propyl together with larger aliphatic and aromatic functions.
WOg4/20062 2 i ~ 7 `1~ 2 PCT~S94/02283 It will also be appreciated that m and n may contain other functional species such as carbonyl, thiocarbonyl, hydroxy, amino, halo and others. Preferred species are carbonyl and thiocarbonyl.
Preferred compounds contain aryl groups in positions Z, E and G. In preferred compounds, Z, E and G are hydroxy substituted aryl, ether substituted aryl, hydroxy substituted heteroaryl, or ether substituted aryl. E and G are preferably substituted aryls or substituted heteroaryls whereby the substitutions are located such that the two aryl rings are conformationally "crowded" out of plane with each other.
Additionally, A is preferably NR or CHz.
Pharmaceutically acceptable salts of these compounds may be used in accordance with the present invention. One having ordinary skill in the art could readily appreciate what salts would be appropriate. Pharmaceutically acceptable salts include, but are not limited to sodium, trialkyl ammonium, potassium, calcium, zinc, lithium, magnesium, aluminum, diethanolamine, ethylenediamine, meglumine and acetate.
Preferred salts are sodium and potassium.
In certain preferred compounds of the present inven-tion A is CH2, NR , S, or O; Bl is NR , O, or CH2; B2 is CO or CS; Z is R4, aryl, heteroaryl, substituted aryl or substituted heteroaryl or D is NR3, O or CH2; E is R5, aryl, heteroaryl, substituted aryl or substituted heteroaryl; F is CO or CS; G is R7, aryl, heteroaryl, substituted aryl or substituted heteroaryl; X is CO or CS; R, R, R, R, R, R, R, R and R
are, independently: hydrogen, lower alkyl or aryl; R5 is lower alkyl or aryl; m is 1-4; and n is 1-4; where n plus m is less than or equal to 5.
In other preferred compounds of the present invention A is CH2, NRl, S, or O; Bl is NR2 or O; B2 is CO or CS; Z is hydroxy substituted aryl, ether substituted aryl, hydroxy substituted heteroaryl, halo substituted aryl; D is NR3 or 0;
E is hydroxy substituted aryl, ether substituted aryl, hydroxy substituted heteroaryl, acyloxy substituted aryl; F is CO or CS; G is hydroxy substituted aryl, ether substituted aryl, W094/20062 PCT~S94/02283 2 1 ~ 7 ~1~ 18 -hydroxy substituted heteroaryl, carboxy substituted aryl; X is CO or CS; Rl, R2 and R3 independently: hydrogen, lower alkyl or aryl; m is 1-2; and n is 1-3; where n plus m is less than or equal to 4.
In still other preferred compounds of the present invention A is NH , CH2 or NR; Bl is NR or O; B2 is CO or CS;
Z is hydroxyphenyl or halophenyl; D is NR3, O or CH2; E is 3,5-hydroxy benzene or 3,5-alkoxy benzene; F is CO or CH2; G is 2-carboxy-benzene, 2-hydroxy benzene, 2,6-dihydroxy benzene, 2-methoxy benzene, 2,6-dimethoxy benzene, or 6-hydroxy benzene-2-carboxylic acid; X is CO; Rl, R2, or R3 are, independently hydrogen, lower alkyl or aryl; m is l; and n is 3.
In certain preferred compounds of the present inven-tion A is NH or CH2; Bl is NH; B2 is CO; Z is p-hydroxyphenyl;
15 D is O; E is 3,5-hydroxy benzene; F is CO; G is 2-carboxy-6-hydroxyphenyl, 2-hydroxy-6-(tetrazol-2-y)phenyl, 2,6-dihydroxy phenyl, 2-hydroxy-1-naphthyl, 2-methoxycarbonyl-6hydroxyphenyl, 2-cyano-6-hydroxy phenyl, and 2-hydroxy-6-(trifluoromethylsulfonamino)phenyl; X is CO; m is l; and n is 3.
As will be appreciated, it is generally the case that one stereoisomer is more biologically active than its enantiomer. It is envisioned that preferred stereoisomerism will be determined for active species and that such preferred compounds will be selected for therapeutic and other uses.
Compounds of the present invention may be synthesized from readily available starting materials by st~n~rd ~er-~n;ques such as by following the basic synthesis set out below. One having ordinary skill in the art may employ other well known synthetic schemes to produce compounds according to the present invention.
Prodrugs such as carbonates and carboxy esters of phenolic OH and NH groups can be prepared by the derivatization of OH and NH groups with acylating agents, such as methyl chloroformate, ethyl chloroformate, isobutyryl chloride, methoxypropionyl chloride, methyl chlorosuccinate, ethyl chlorosuccinate and benzoyl chloride, for example.
21~74~ 2 W094l20062 PCT/US94/02283 Additionally, prodrugs of compounds which contain a carboxylic acid can be prepared by derivitazation with alkylating agents, such as methyl iodide or acetoxymethyl chloride.
Reaction Scheme I provides syntheses for producing compounds according to the present invention including the use of a cyclic carbonyl or a heterocyclic carbonyl such as the seven membered lactam shown as a starting material. For example, a lactam can be benzylated with base in tetrahydrofuran to protect the nitrogen functionality. It is then reacted with base and phenylselenyl chloride followed by sodium periodate to yield the unsaturated lactam. Oxidation with osmium tetroxide followed by benzoylation yields the hydroxy benzoate shown. Further reaction with trifluoromethanesulfonic anhydride and sodium azide provides the anti azido ester which can be reduced with, for example, lithium aluminum hydride. Reaction with a carboxylic acid substituted with a Z functionality yields the amide. Further reaction with a GFE carboxylic function followed by deprotection provides the family of ester/amides.
2157~112 -20-O
_~z = ~ Z ti `C
T ~ T
O t_ ~Z= e_~
.' T ~,, o T
m . .
~ m ~:
Q IL Q I
Z Jl-- ~ O
C~ 11 -- ' --C
W094/20062 ~i 7 11~ PCT~S94/02283 Scheme II provides a synthesis scheme for producing compounds according to the present invention including the use of an enol ether lactam such as the azepinone shown as a starting material. For example, an enolether lactam can be benzylated with base in tetrahydrofuran to protect the nitrogen function. It is then hydrolyzed with acid and reacted with sodium nitrite in acetic acid to form the oxime. Catalytic hydrogenation in the presence of acetic anhydride gives the acetamide, which can be reduced with, for example, sodium borohydride and hydrolyzed to the syn (or anti) aminohydroxy lactam. Reduction with, for example, lithium aluminum hydride and reaction with a carboxylic acid substituted with a Z
functionality yields the amide, which can be further reacted with GFE carboxylic function and deprotected to provide the family of ester/amides.
WO 94noo62 rCT/US94/02283 2 1 5 7 ~c~jeme II
OEt OEt ~O
1)KOtBu ~ HCUH20 ~
~ N ~ o 2)PhCH28r Ph N~ ~h Y.Tamuraetal., Chem.Pharm.Bull., ~(3).523-8,529-34(1971) 1 ) NaBH4 ~2)HCUH20/EtOH
NaNO2 ~ NOH H2/RaneyNi ~ NHCOCH3 3) NaOH
ACOH/Ac20 ~ N ~ O AcOHlAc20 ~ O
Ph Ph H ~OH Z-COCI OH
NHz LAH (~ IH2 Et3N/DMAP(~;~NHCO--Z
Ph Ph Z-C-N ~N Ph G-F-E-COCI
Et3NtDMAP lc_E-F-G H2 olC_E-F-G
~ ~NHCO - Z
G-F-E-COOH ( ~ NHCO - Z Pd(OH)2;C H
DCCIDMAP
W094/20062 21~ 2 PCT~S94/02283 Reaction Scheme III provides methods for producing compounds according to the present invention including the use as a starting material of the previously mentioned syn aminohydroxy lactam. Oxidation of the alcohol using, for example, oxalyl chloride, dimethylsulfoxide, and triethylamine (Swen oxidation) provides the Keto intermediate which is treated with HONH2-HCl followed by reduction using, for example, RaNi (Raney nickel) catalyst and hydrogen affords the amino-amide. Reaction with a GFE carboxylic acid function followed by deprotection provides the family of diamides.
215 7 ~ 24 - PCTIUS9~/02283 Scheme m ~OH O
(~NHCO--Z oxalyl chloride ~NHCO Z 1) NH20H-HCI
DMSO, Et3N, -65C NJ 2) RaNi. H2 Ph Ph G-F-E-COCI
~NH2 Rm Et3N/DMAP NHCO-E-F-G
(;;)~NHCO{~ or O~ NHCO--Z
G-F-E-co-N~g N
Ph Ph NHCO-E-F-G
~NHCO Z
Pd(OH)2/C H
W094eO062 ~ 3 5, ~12 PCT1594/02283 Scheme IV provides a synthesis scheme for producing compounds according to the present invention including the use as a starting material of a cyclic olefin as shown.
Epoxidation with peracetic acid followed by reaction with sodium azide affords the anti azido alcohol, which is O-protected and reduced to the aminoalcohol ether. Reaction with a carboxylic acid substituted with a Z functionality yields the amide, which is O-deprotected and reacted with a GFE carboxylic function to provide the family of ester/amides.
WO 94/20062 rCT/US94/02283 2 1 5 ~ 26 -t~ T
=:E O ~
IL -~3 ~
m ~ ~0~
~0 s _ c ~
_ c T
T
Z~
, E m ~
z~
~ o T T
0~ ~
N _ T
~ o U~f~O
z 5 0 C~ ~ Z`'-o , ~ r~l ~0 W094/2006~ 2 1 ~ 2 _ PCT/US94/02283 Scheme V provides a syntheses for preparing compounds according to the present invention including a synthesis scheme for producing 6, 7 and 8 member cyclic and heterocyclic groups including Bl and D with stereo specific attachment, including the use as a starting material of the unsaturated aldehyde.
For example, tin-mediated condensation and subsequent cyclization of the aldehyde with an isothiocyanate affords the oxazolidine thione, which can be reduced with lithium aluminum hydride, ozonolyzed, and further reduced with, for example, sodium borohydride to the diol. Mesylation with methanesulfonyl chloride and base followed by ring closure with benzylamine affords the azepine, which is hydrolyzed with an acid, for example, hydrochloric açid. The resultant anti aminoalcohol is reacted with a carboxylic acid substituted with a Z functionality to give the amide, which is further reacted with a GFE carboxylic function and deprotected to provide the family of ester/amides with predictable stereochemistry at the positions of attachment of B1 and D to the ring system.
WO 94/2006~
21~7 4~ -28 ô
CL
o~ ~ I I
Z
m ~<O~
--~--LIJ_~
~O~cn z J Z
~ .
T I oN
> ,_ Z O ~
O=~ c ~ ~ U e~
0=( ~ 5 O ~<O~
N~ 3~ ~> O
O ~ ,~
Z N m , N
3~ C`' ~.) C~
0~ ~ (~ _ ~ Z_~m O O
0~
W094/20062 2 1 ~ 7 ~ 1 2 The reactions of Scheme VI provide a synthesis scheme for producing 6, 7, and 8 membered cyclic and heterocyclic groups including Bl and D with stereo specific attachment including the use as a starting material of N-carbobenzyloxy asparagine. For example, CBZ-asparagine is reacted with bis(trifluoroacetoxy) iodobenzene to give the mono-protected - diaminoacid, which is differentially protected with di-t-butyl dicarbonate and reduced with, for example, borane/tetrahydrofuran. Oxidation to the aldehyde and condensation with an unsaturated organometallic affords the diprotected diamino alcohol, which gives the terminal tosyloxy compound after hydroboration/oxidation and treatment with toluene sulfonyl chloride. Removal of the butoxycarbonyl is accomplished by acid treatment, for example, formic acid or trifluoroacetic acid, followed by removal of the amine-protecting group, which can be removed with hydrogen.
Selective reaction with a carboxylic acid substituted with a Z
functionality followed by treatment with benzyl chloroformate and base gives the protected amide, which is further reacted with a GFE carboxylic function and deprotected to provide the family of ester/amides with predictable stereochemistry at the positions of attachment of B1 and D to the ring system.
2 1 5 ~ l 1 2 Scheme Vl H~N$~ ~_NHcez DMAP NEI ~~~ ez NHCeZ ~ ~ S wcr~
HN ~aM5CI , HN
80C n=0-2 80C 80C
l) BBN
2) H~02 3) Tsa TsO/~ z) tase ~ NHCeZ ~ Pd~OH) ~ N
Azapine 1.
Int~..c~ with alk...~ti~ synthcsis shown on nc:a shccet SUBSTITUTE SHEET (RULE 263 W094/20062 21~ ~ k ~ ~ PCT/US94102283 Scheme VII provides a syntheses for producing compounds according to the present invention including a synthesis scheme for producing 6, 7, and 8 member cyclic and heterocyclic groups including Bl and D with stereo specific attachment including the use as a starting material of phthalimide alkyl aldehyde. for example, the aldehyde can be reacted with methyl isocyanoacetate in the present of gold ferrocene catalyst to give the oxazolidine, which is hydrolyzed to the diaminohydroxy ester salt with for example, hydrochloric acid. Base mediated cyclization affords the lactam, which can be reduced to an aminohydroxy compound. Reaction with a carboxylic acid substituted with a Z functionality followed by treatment with benzyl chloroformate and base gives the protected amide, with is further reacted with a GFE carboxylic function and deprotected to provide the family of ester/amides with predictable stereo chemistry at the positions of attachment of B1 and D to the ring system.
.'O 94/20062 2~7~12 -32-Scheme Vll O Gold F~ ucc~c O
~N~r~ C ?~'CH,CO2CH3 [~ ~; O
O CHO O ,~
H3CO2C ~N"
n=0-2 1) 6NHCI
2) MeOH, HCI
OH `~
(~ .,NH2 MeOH,K2CO3 yH2-HCI
~/ ~ HCI-H2N ~C02CH3 N ~o OH
LAH
OH 1) ZCOCI OH
~'EL3, D~ N~r z N CBZ
H
Azapinc 1 G-F-E-COCI
~ ~Et3, DMAP
E--F-G
E--F-G O~
o H2, Pd(OH)2 _~ H
~,... ~N~rz lFA.ELOH C8Z
W094/20062 2 ~ ~ 7 ~ ~ 2 rcT~sg~/02283 Scheme VIII A provides a synthesis scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including Bl and D with syn attachment including the se of the protected cyclic ketone as starting material. For example, the ketone is deprotonated and the enolate aminated to afford the butoxycarbonylamino ketone which can be stereo specifically reduced with, for example, sodium borohydride to the syn aminoalcohol. Reactions with a carboxylic acid substituted with a Z functionality yields the amide. Further reaction with a GFE carboxylic function followed by deprotection provides the family of syn ester/amides.
WO 94/20062 PCTtUS9 1tO2283 7 ~ 1 2 34 o I
N m _~ ,O~ZT
Z
m O m ,~ I
._ T
O-~Z~ +' TZ
~ ~0--~Z ~ ~
T _ O
O~Z~ ~
+ + ~
,~ ~ X o X ,.
~ o U U D
-- o~Z~ o o z -W094/20062 21~ PCT~S91/02283 Scheme VIII B provides a synthesis scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including B1 and D with anti attachment including the use as starting material of the syn hydroxyamide from Scheme VIII A. For example, the syn hydroxyamide can be inverted to anti hydroxyamide by treatment with carboxylic acid, such as acetic acid, in the present of triphenylphosphine and diethylazodicarboxylate followed by treatment with sodium methoxide. Reaction with a GFE
carboxylic function followed by deprotection provides the family of anti ester/amides.
WO 9~/20062 PCT/US9~/02283 21~7~12 -36-~
X X IZ ~, U U O ~ ZI Nl C
U
Q
Z
T~ a~ ., O ~ JQ -- o o D D C~
X ~ T
U O
~N
~ IZ '~
o ~Z ~ 11 u _ E
C~
W094120062 21~ ~ ~12 PCT/US94/02283 Scheme VIII C provides methods for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including Bl and D with anti attachment including the use as starting material of the syn hydroxyamide from Scheme VIII A.
For example, the syn hydroxyamide can b~ inverted to anti amino amide by treatment with trifluoromethane sulfonic anhydride and sodium azide followed by reduction with, for example, tin (II) chloride. Reaction with a GFE carboxylic function followed by deprotection provides the family of anti diamides.
WO 94/2006~
21 ~! 112 -38-_ I
r ~
o Z.. ~ZT c~l C ¦
U U U Z"
~ ' _ o t~l N
m .q Z~Z Z
D O o ~ ~ _ N ~ N
_~ a ~J
W094/20062 2 ~ 3 PCT~S91/02283 Scheme VIII D provides a synthesis' scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including Bl and D with syn attachment including the use as starting material of the anti hydroxyamide from Scheme VIII B. For example, the anti hydroxyamide can be inverted to syn amino amide by treatment with trifluoromethanesulfonic anhydride and sodium azide followed by reduction with, for example, tin (II) chloride.
Reaction with a GFE carboxylic function followed by deprotection provides the family of syn diamides.
~'O 94/20062 N
O
+ <, 3 U O X ~ Z
,, ~, U
N
T
a I_~z~ . Z
N
O
~ o ~ E
W094/20062 21~ 7 41~ PCT/US94/02283 Scheme VIII E provides a synthesis scheme for producing compounds according to the present invention including producing 5 and 6 member cyclic and heterocyclic groups including Bl and D with anti attachment including the use of the protected cyclic epoxide as starting material. For example, the epoxide is opened with ammonia to provide the anti aminoalcohol or with azide to provide the azido-alcohol.
Reduction of the latter with, for example, triphenylphosphine provides the amino alcohol. Reaction with a carboxylic acid substituted with a Z functionality yields the anti hydroxyamide. Further reaction with a GFE carboxylic function followed by deprotection provides the family of anti ester/amides.
WO 94/20062 rCT/US91/02283 2 1 ~ 7 4 1 2 -42-D
X m Ll O U --, GZ I U~l / e x Z I
~ Z~
N ~
m o Z--,G n ~ C~L"_ e ~ ~ ~
D <~ --3 z ~
~, o ~ I o m N ~ z e ~J5 D
Z~~\ n +~
~
S O~z~
WO9ql20062 21~ PCT/159~;02283 Scheme VIII F provides a synthesis scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including Bl and D with syn attachment including the use of the anti hydroxyamide from Scheme VIII E as starting material. For example, the anti hydroxyamide can be inverted to syn hydroxyamide by treatment with carboxylic acid, such as acetic acid in the presence of triphenylphosphine and diethyl azodicarboxylate followed by deprotection provides the family of syn ester/amides.
2~^~7~
o I ~, N
~-) XO T
~ U --_~
~~
X
U
N ~
0~ 0 ~
w O D
X tL
u m O
'~QZ E
E
-C~
W094/20062 2 ~ 12 PCT/US9~/02283 Scheme VIII G provides a synthesis scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including B~ and D with syn attachment including the use of the anti hydroxyamide from Scheme VIII E as starting material. For example, the anti hydroxyamide can be inverted to syn amino amide by treatment with trifluoromethanesulfonic anhydride and sodium azide followed by reduction with, for example, tin (II) chloride.
Reaction with a GFE carboxylic function followed by deprotection provides the family of syn diamides.
2 1~ l 4 12 O
+ <~31 -~ZI
U ,, U
O ~ ~ _ tO ~3 o TZ X~Z ~ Z
N ~_ n --I ~ ~ CL
c~ I ~ _ X S
r~ , U
N
E I E E
U, W094/20062 215 ~ ~ ~ 2 PCT/US94/02283 Scheme VIII H provides a synthesis scheme for producing compounds according to the present invention including a synthesis scheme for producing 5 and 6 member cyclic and heterocyclic groups including the use of the syn hydroxyamide from Scheme VIII F as starting material. For example, the syn hydroxyamide can be inverted to anti amino amide by treatment with trifluoromethanesulfonic anhydride and sodium azide followed by reduction with, for example, tin (II) chloride. Reaction with a GFE carboxylic function followed by deprotection provides the family of anti diamides.
WO 9~/20062 2 1 ~ ~ 4 1 2 48 ~I tL
Z .~, I
</~
~ x I .. ~
U U X
o ~ U
o o OI
a ~ t~
~ C D
Z'--~ Z
~ O
D _, Q~ C
U
~ a T--~ ~ ~ N z C~
W094/20062 21~ ~ ~1~ PCT/US9S/02283 Scheme IXA provides a synthesis scheme for producing intermediate GFE carboxylic acids including the use of 4-bromobenzoic acids as starting material. For example a 4-bromobenzoic acid can be esterified by treatment with carbonyldiimidazole followed by an alcohol such as tert-butanol. The ester is treated with n-butyl lithium followed by an addition of N, N-dimethyl formamide to give the aldehyde ester, or by addition of carbon dioxide to give the acid ester, which is converted to the acid chloride ester with oxalyl chloride. The ester can also be reacted with n-butyl lithium and a benzaldehyde to give the diphenyl carbinol, which is oxidized with, for example, chromic acid, and deprotected to provide GFE carboxylic acids. The ester can be reacted with n-butyl lithium and benzoyl chloride to give the diphenyl ketone, which is deprotected to also provide GFE carboxylic acids.
21~12 .so.
Scheme rx A L) n-suLiO c~c~2~
Er~CO2H ,, ~,~C02R --2) CO2~CO2R
~ (COC~).
CIOC~CO2R
X 1) ~ ~CO R
O O
~COzR ~CO2H
Er~ 1) B .I; ~CO2R
o ~CO2H
SUBS~lTUTE SHEET (RULE 2~
2~7412 WO9~/20062 PCT/US91l02283 Scheme IXB provides a synthesis scheme for producing intermediate GFE carboxylic acids including the use of either
4-bromobenzoic esters from scheme IXA or benzyl alcohols as starting material. For example, a 4-bromobenzoic ester can be treated with n-butyl lithium and phthalic anhydride to afford the 2'-carboxybenzophenone ester, which is protected with, for example, benzyl alcohol esterification at the 2' position and deprotected at the other ester position to provide GFE
carboxylic acids. In an alternative, preferred method, the alcohol is coupled with the acid chloride and treated with one equivalent of nBuLi which provides, upon rearrangement, the hydroxy ketone. Oxidation to the acid is effected in a two-step process using first pyridinium dichromate (PDC) or TEMPO
(2,2,6,6-tetramethyl-1-piperidinyloxy) followed by tetrabutylammonium permanganate (nBu4MnO4) or sulfamic acid (NH2SO3H) and sodium chlorite (NaClO2). The resultant acid is protected, for example, by conversion to the benzyl ester and then deprotected at the other ester position to provide the family of GFE carboxylic acids.
21~7 4~ 2 ~O 94/20062 PCT/US94/02283 Scheme IX B - 52-/=\ 1) BuLi ~ protect ~ 2) 5~ Iyl/J x ~CO2R
R'02C O R'02C O
~ dep~tec~, 5~CO2H
HO HO~
Br ~CO2R []
CIOC ~ CO2R ,KOtBu X
2) nBuLi HO2C O R'02C O
~ protect ~CO2R
R'02C O
depl~)tect ~CO2H
2~5 ~J~1~
W094/20062 rcTluss4lo2283 Scheme IXC provides a synthesis scheme for producing GFE carboxylic acids including the use of bromobenzenes as starting material. For example, a bromobenzene can be treated with n-butyl lithium and the acid chloride ester from scheme IXA to afford the benzophenone ester, which is deprotected to provide GFE carboxylic acids. The bromobenzene can also be treated with n-butyl lithium and the aldehyde ester from scheme IXA to give the diphenylcarbinol ester, which is either deprotected directly, oxidized with, for example, chromic acid and deprotected, or reduced with, for example, hydrogen and deprotected to provide the family of GFE carboxylic acids.
2157~1~?
WO 94/20062 PCT/US94/02283 _ Scheme ~X C
~r 1) ~'-r ~ CO R ~COzR
tCy~utc~L ~CO2H
L~.U...,L ~`CO2H
2) ~CO R ~CO2R
O ~ \
~CO2R ~CO2R
d~y~lt~ deprotect ~CO2H ~CO2H
S~I~UTE S! IEE~ (RULE 2~J
2157~12 WO9~/20062 PCT/US91102283 Scheme IXD provides a synthesis scheme for producing compounds according to the present invention including the use of halobenzenes, for example, bromobenzenes, and heterocyclic or cyclic compounds substituted with B1-B2-Z and DH, which are described in other schemes, as starting materials. For example, a bromobenzene can be reacted with a 4-heterobenzoate ester, in dimethyl formamide/triethylamine with heat or copper catalyst to give GFE acid chloride. This can be treated with heterocyclic compounds substituted with B1-B2-Z and DH in the presence of tetrahydrofuran/triethylamine to provide this family of balanoids.
PCT/US9~/02283 21~7~
Scheme ~X D
Arm 1 synthesis (continued):
DMF/Et3N
,Br HQ~ DMFlEt~N/Cu catalyst Q=O,~H,S
~3,a~ Ester ~ ~3`cocl HD~ B2--Z y ~CO
D~B, - B2--Z
Et3N/THF A
W094/20062 21~ ~ 4 ~ 2 rcTlus94lo2283 Scheme IXE provides a synthesis scheme f~or producing compounds according to the present invention including the use of aromatic or heteroaromatic halides protected, hetero-substituted aryl or heteroaryl acids or alcohols, and heterocyclic or cyclic compounds substituted with Bl-B2-X and DH, which are described in other schemes, as starting materials. For example, a bromoaromatic can be reacted with a protected 4-hetero-substituted aryl acid or alcohol in the presence of transition metal catalyst, for example, copper, and base to afford protected GFE acid or alcohol, which can be deprotected and treated with, for example, phosphorous penta chlorides to provide GFEX chloride. This is reacted with heterocyclic or cyclic compounds substituted with Bl-B2-Z and DH to provide another family of balanoids.
2 1 ~
- s8 -Scheme IX E
Cu catalyst G Br+ HF E XOR ~ G F E--XOR
or Pd(0) or Ni (0) E~3N
d~plotccL PCIS
G F E XOH
HD B~B2Z G F E--X\
~. D B1 Bz-Z
G--F--E XCI , \1~
E-,~IT~ / \
~A~)n W094/20062 2 1 5 7 ~ ~ ~ PCTIUS9~102283 Scheme IX F provides a preferred method for the preparation of the GFE-CO2H. Intermediate aldehyde is first protected, for example, as a cyclic acetal, which is then treated with nBuLi and DMF to afford aldehyde 1. Aryl bromide, is treated with n-butyl lithium and then aldehyde I to afford the alcohol. Oxidation using, for example MnO2 followed by acidic hydrolysis of the acetal gives the ketoaldehyde.
Oxidation to the acid using, for example, sodium chlorite and sulfamic acid, followed by the appropriate deprotection sequences affords GFE-CO2H.
wo 94/20062 PCT/US94/02283 2 1 5 ~ 60 -Scheme IX F
o o I Y I X~
/=1=\ 1) nBuLi ~/ ~ ~/~ 1) oxidation 2) 1 C~~ c/J~ 2) acid, H20 ~/~ 1) o~id~ion ~f/
2)protection /~1~
CO2R3)deprotection / CO2H
x2 x2 W094/20062 21~ 7 412 PCT~S9~/022~3 -Scheme IX G describes a preferred method for the preparation of GFE-CO2H where E is substituted with two OH or OR groups. Protected bis-phenol is treated with n-butyl lithium followed by aldehyde (prepared in Scheme IX F) provides, after oxidation with, for example, MnO2 the ketone intermediate. Following deprotection, the primary alcohol is oxidized, using, for example MnO2 followed by sodium chlorite/hydrogen peroxide. Hydrolysis of the acetal followed by treatment with MnO2, KCN, acetic acid and the alcohol R'-OH
provides the desired GFE-CO2H.
A detailed scheme for benzophenone synthesis is shown in Scheme IX H.
wo 94/2006~ - 62- PCT/US94/02283 Scheme IX G
O OCH3 y L~ l)nBuLi ~ OCH3 ~/r CHO O~ OCH3 o 3) oxidation o o OCH3 1) deprotection ~ H30+
2) oxidation ll o ! _ .
3) esterification ~ ~ / ~ CO2R
~ ~
o OCH3 O OCH3 Y ~ 1 1) KCN, AcOH /Y
~/~ ~MnO2, R'OH
CHO O/~2) dep~otection ~ ~ C02R' ~ CO2H
WO9~/2006~ 2 1 ~ 7 ~ l 2 - PCT/US9~/02283 Scheme IX H
Synthesis of MOM-Protected Benzophenone CO.M~ CO.Me ~OH
~3~ MOMCI ~ LAH, T~ ~
HO OH MOMO~OMOM MOMOJ~OMOM
OT~DMS
TBDMSCI r e ~
1~/10~10~0~10~1 HO nnO BnO
K,C03,BnBr ~ 1,3~ ,l).u~ H7~-C
CHO ~ PhMe,~ ~ o O~
HO hlOMO MOMO
f~q N.-lH, MOMCI ~ "BuLi, C6HI2 ~CHO
n ~MF ~
OTBDMS MOMlo hlOhlO~OTl~DMS
+ ~, I ''BuLi,0''C ~<
MOMO OhlOlM O J .1 ~
WO 9~/20062 PCT/US9 ~/022~3 2 1 ~
~ 64 ~
MOMO MOMO~,OMOM MOMO~OTBDMS
¢~0 OT~DMSh-- OMOM MnO~, CH,CI
o J "BuLi, 0"C. THF
O
MOMO~ OTBDMSMOMO~ OH
MOMO o ~ MOMO o ~ \\
~< TBAF, THF ~ MnO., CH.CI, OMOM > ~ OhlOM
O O
MOMO ~_~CHO hlOhlO~CO,hle MO~ KOH, 1, MO~
W~ OhlOlM MeOH. 0"C W~ OM()hl O O
NaCI02. HtO~ 18% H~SO~
N.aH2PO.~, MeCN-H.O a~l;orbe~ n SiO2 CH2( 12 MOh~O CO~H hlOlMO CO~Me MO~ MO~
~ OhlOhl W~CHO
WO 94/2006~
~1 J ~ ~ _ ~ PCT/US94/02283 ~,HO MOMO~,OMOM 'Blll i O"C B~ U~OTBDMS
~ ll l OMOM
OOTBDMS ~
o MlOhlO~OTBDMS MOMO ~--OH
MnO.. CH2CI- n,~ TBAF B~
W~ OMOM W~ OMIOMI
O O
MOMO~ ~CHO M~ lO~ CO2H
BnO O \ \\ BnO ~ \\
MnO-. CH-a2 ~< NsClO2, NaH2PO1 ~ <
H,02,MeCN-H.O l~'~~ OMOM
O O
2~ 2 - 66 -Scheme X provides means for produciAg compounds according to the present invention including the use of heterocyclic or cyclic compounds substituted with B1-Bz-Z and DXEFG where F is C=o, which are described in other schemes, as starting materials. For example, a heterocyclic or cyclic compound substituted with B1-B2-Z and DXEFG where F is C=O can be treated with a reducing agent such as sodium borohydride to afford balanoids where F is CHOH, or it can be treated with a sulfurizing agent, for example, phosphorus pentasulfide, to provide the family of balanoids with B2 and/or F equal to C=S.
~ro 9~/20062 21 S 7 412 PCTIUS9~102283 Scheme X
;~ G 1l HE~ G
I E
m=0-3 P4SIOor Lawesson's reagent 111 E~G
DC ll3 B~ C_ z A
X~,X2,X3 in~cpe~ ly = O or S
2 1 J7 '~12 - 68 -Scheme XIA provides a synthesis scheme for producing compounds according to the present invention including the use of heterocyclic compounds substituted with B1-B2-Z and DXEFG
where A is NH, described in other schemes, as starting materials. For example, a heterocyclic compound substituted with Bl-B2-Z and DXEFG where A is NH can be treated with an alkylating agent, sulfonylating agent, or acylating agent, for example acetyl chloride, in the presence of base to provide the family of balanoids with a substanted nitrogen.
In an alternative approach, Scheme XI B, the R group can be appended at an earlier stage in the synthesis using appropriately protected intermediates. For example, heterocyclic intermediate can be treated with an alkylating agent, sulfonylating agent or acylating agent, in the presence of base to provide the intermediate where Rl is not hydrogen.
2 PCT/US9 ~/02283 Scheme XI A - 69 -G--F--E--X-D~--B2--Z G--F--E--X-D 31-3z--Z
n( )m Et3N n(~ )m H Rl m=1-4,n=14 Rl=lower alkyl, aryl, or JR2 where m+n~or=5 J=CO,CN, or SO2 and R2=lower alkyl, aryl, alkylamino, or alkoxy Scheme XI B
HO~B1 -B2 ~Z ~B1 -B2 ~Z
N Et3N N
H Rl as described )--( in Scheme I Rl 2157~1~
WO9~12006' PCT/US94102283 Scheme XII provides a synthesis scheme for producing compounds according to the present invention including the use of cyclic or heterocyclic compounds substituted with B1-B2-Z and DXEFG where X is C=O, which are described in other schemes, as starting materials. For example, a heterocyclic compound substituted with B1-B2-Z and DXEFG where is C=O can be treated with an alkylating agent, for example dimethyl sulfate and an alcohol, such as methanol, to give the intermediate ketal.
This can be reacted with an organometallic, for example, the lithium salt of diethyl malonate, to provide the family of balanoids in which x is C=CR3R4. The ketal can be reacted with a primary amine, for example, butylamine, to provide the family of balanoids in which X is C=NR2.
WO 94/2006~ 7 - - 71 - ~ cL ~ PCT/US911022S3 X--\
E /
U~
a X E X E
O~
G U U
C~
oN
_ o C~.~
m c E ~X E/
O=~
~ U
~VO91/20062 PCT/US91/02283 57 41~ Scheme XIII provides syntheses for producing compounds according to the present invention including the use of GFE
carboxylic acids and cyclic or heterocyclic compounds substituted with B1-B2-Z and DH or B,H and DXEFG, which are described in other schemes, as starting materials. For example, a GFE carbinol can be treated with mesyl chloride and an amine base, such as triethylamine, followed by treatment with an iodide source such as sodium iodide to afford a GFE
methyl iodide. This can be reacted with a cyclic or heterocyclic compound substituted with Bl-B2-Z and DH in the presence of base such as sodium hydride to provide the family of balanoids in which X is CH2.
The cyclic or heterocyclic compound substituted with B,H and DXEFG can be reacted with a Z sulfonyl chloride, for example, benzenesulfonyl chloride, in the presence of base to provide the family of balanoids in which B2 is SO2.
N
o~ I
U
U ~ ' ~,u z ~ m E
s z I ' X~s X
u ~ U Z
E
U~ .
21~7~ 74 -Scheme XIV A provides a synthesis scheme for producing intermediates which are heterocyclic compounds substituted with Bl=NH2 and D=OH and in which A is oxygen. For example, a cyclic diene such as cyclopentadiene can be treated with a peracid such as peracetic acid and the epoxide opened with an amino species, for example, (dibenzylamino) dimethyl aluminum, followed by butyldimethylsilyl chloride to obtain the protected amino alcohol. This can be ozonolyzed and reduced to the diol, which is treated with tosyl chloride to give the chloro tosylate. Treatment with bis(trimethyltin)oxide or preferably potassium superoxide and a crown ether, for example, 18-C-6, provides the chloroalcohol which is ring closed on treatment with a strong base, such as methyllithium or butyllithium to afford the protected heterocyclic compound, which is deprotected to heterocyclic compounds substituted with B~=NH2 and D=OH and in which A is oxygen.
~'O 91/2006' 5 2 1 5 7 4 1 ~ PCT/US9 ~/02283 Scheme XIV A
1) Bn2NAIMe2 NBn2 Ac02H ~ 2) TBDMS-CI ~
1-3 1-3 ~ 'OTBDMS
1) 03 OTBDMS OTBDMS
2) NaBH4 Ts-CI .
HO~OH ~ Cl ~OTs NBn2 NBn2 2) BuLi, toluene, heat TBDMSOs NBn2 [H] TBDMSOs NH2 n(i~)m n(~ )m F- HOs NH2 n(~ )m 2 1~7 ~ ~/20062 - 76 - PCT/US9~/02283 Scheme XIV B provides a synthesis scheme for producing intermediates which are heterocyclic compounds substituted with B1=NH2 and D=OH and in which A is sulfur. For example, a cyclic diene such as cyclopentadiene can be treated with a peracid such as peracetic acid and the epoxide opened with an azide species, for example, sodium azide, followed by t-butyldimethylsilyl chloride to obtain the protected amino alcohol. This can be ozonolyzed and reduced to the diol, which is treated with mesyl chloride to give the dimesylate and ring closed with, for example, bis(trimethyltin) sulfide or preferably, lithium sulfide and an amine base such as triethylamine. Reduction of the azide and deprotection occurs on treatment with lithium aluminum hydride to give heterocyclic compounds substituted with Bl = NH2 and D = OH and in which A
is sulfur.
Compounds where A is SO2 can be prepared from compounds where A is sulfur by treatment with an oxidizing agent, for example, peracetic acid,followed by deprotection to provide compounds wherein A is SO2 (Scheme XIV C).
WO 94/20062 21 S 1 ~ ~ ~ PCT/US91/02283 Scheme XIV B
1) NaN3, NH4CI, CH30H/N3 Ac02H ~< 2) TBDMS-CI
1-3 1-3 ~ "OTBDMS
1) 03 OTBDMS OTBDMS
2) NaBH4 . Ms-CI
HO~OH ~ MsO ~OMs N3 NBn2 1 ) Li2S, Et3N, CH30H HO NH2 2)LiAlH4 S
-'O 9~/~006' Scheme X~V C - 78 - `-215~
/ /
___ 1) RCO3H ~ A
S~ )m 2) De~rut~c~ion n( ~ ~ )m O O
~ro94/20062 215 ~ ~1~ PCT/US94/02283 Scheme XV provides a synthesis scheme for producing compounds according to the present invention including the use of Z sulfonyl chlorides, such as benzenesulfonyl chloride, and cyclic or heterocyclic compounds substituted with Bl=NH2 and DH, which are described in other schemes, as starting materials.
For example, a Z sulfonyl chloride and a cyclic or heterocyclic compound substituted with Bl=NH2 and DH are combined in the presence of base to afford the sulfonamide, which is reacted with GFEX halide to provide the family of balanoids in which B
is N and B2 is SO2.
WO 94/2006' PCT/US93/022S3 2 ~r'J - 80 -Scheme XV
ol o D NH2 Cl--S--Z D N--S--Z
m( ~ ) n Et~N m( ~ ) n G F E X~ H ¦¦
G - F E--X--Cl D N--S--Z
m( ~A~ ) n G--F E X~ R
D N_S--Z
K2CO3, R2X \~ ( o m( g~A~ ) n W094/20062 2 1 5 ~ ~ ~ ~? PCT/US9S/02283 Scheme XVI provides a synthesis scheme for producing compounds according to the present invention including the use of ketones, for example, acetophenone, and azepinediones, such as 1-benzylazepin-2,4-dione, as starting materials. For example, the ketone can be brominated with bromine in acetic acid and protected with ethylene glycol to afford the bromomethyl ketal, which is added to a base-treated solution of the azepinedione to give the alkylated azepinedione. This is reduced to the hydroxyazepine with, for example, lithium aluminum hydride, and reacted with base and GFEX halide, which is described in other schemes, then deprotected to provide the family of balanoids where Bl is CH2, Bz is C=O, and D is O.
These compounds can be reacted with, for example, phosphorus tetrasulfide, to provide the family of balanoids where Bl is CH2, B is C=S and D is O.
WO 94/2006' 2 1 5 ~ 82 -Scheme XVI
--'6Z Br2, HOAcBr~_~Z HOCH2CH20H, Br 7~
O o pTsOH, Benzene, O J
Dean-Stark Trap ~,~ 1 ) NaH ~ LIAIH~
G--F E X
OH O
(~<Z 1.) NaH (~<Z
N \J 2-~ Cl X E--F--G N \_l Rl R
G--F E X
1.) PPTS, Acetone H20 , (~' P4S, o N
G--F E X\
o (~Z
N S
-'09~/20062 ~ 2 PCT~S9~10228 Scheme XVII provides a synthesis scheme for producing compounds according to the present invention including GFE
carboxylic acids, which are described in other schemes, and cyclic or heterocyclic groups substituted with OH and BlB2Z, also described in other schemes, as starting materials. For example, a GFE carboxylic acid can be treated with oxalyl chloride and N,O-dimethylhydroxylamine to afford the methoxymethyl amide, which is reacted with a methyl organometallic to give the methyl ketone. This is then deprotonated with base and reacted with the product from treatment of a cyclic or heterocyclic group substituted with DH
and BlB2z with tosyl chloride to provide the family of balanoids with D as CH2 and X as C=O.
WO 9~/2006~
- PCT/US9~/022~3 2157~12 Scheme XVII
?
) (COC1~2/D~
CH2Cl2 ~'.~"
G F--E--CO2H ~ G F--E--CON(OMe)(Me) 2) MeNHOMe M=Li, MgBr G F E--COCHa HO B~ B2Z TsCI TsO B~ B2Z
(~)n ~ (~) LiN(SiMe3)2 G--F E--COC~Bl B2Z
(b~A~)n W094/2006~ 215 i ~ CT/US94/02t~3 Scheme XVIII provides a synthetic scheme for compounds of the invention where K is not equal to H. Oxidation of the alcohol using, for example, oxalyl chloride and dimethylsulfoxide (Swern oxidation) followed by addition of an organometallic reagent, for example, the complex of trimethylaluminum and methyl magnesium chloride to afford the tertiary alcohol. This intermediate is converted using the Scheme I to compounds of the invention.
wO 9~/2006~
Scheme XVIII
2157~12 HO B1 - B2--z 1 ) oxalyl chloride HO ~ B1 - B2--Z
)~ DMSO, Et3N )~
A/~ 2) Me3AI, MeMgCl Q\A/Q
as described Targets where K = CH3 in Scheme I
~094/2006~ 21 S 7 41~ PCT/US94/02283 Scheme XIX provides a synthetic scheme for compounds of the invention where the group G is substituted with an alkoxycarbonyl group. Compounds wherein G is substituted with a carboxy group are treated with an alkylating agent, for example, methyl iodide, and a base such as sodium carbonate to provide the target compounds.
WO 9~/20062 - 88 - PCT/US9~1/022~3 ~cheme ~lX
C~X~
X D K B IB2Z R-halo ~A~ )m K2CO3, DMF
OH
Y I X
X~
x2 DK B IB2Z
n(~ )m where R = alkyl, substit-1tPd alkyl wo 9~,2006? 2 1 S 7 4 1 2 PCT/US9~/02283 Scheme XX provides a synthetic route for compounds of the invention where G and/or E residues are substituted with acyloxy groups. Target compounds, which possess one or more hydroxyl groups on G or E is treated with an acylating agent, for example, acetyl chloride, ethyl chloroformate, and the like, in the presence of a base such as pyridine or triethylamine to provide the target compounds.
WO 94/2006 2 PCTIUS9 ~/022~3 Scheme XX ~ 90 ~
lS~ 41~ o OH HO~ X 1 ) _1l D K B IB2Z R Cl pyridine ~A~ 2) dep~~ L
~O~o~X~ ~
o~L O~ D~ B~B2Z
n(~A~ )m ~'O9~/20062 21~ 7 ~12 PCT/US9~/022X3 Scheme XXI provides a synthetic scheme for the preparations of compounds where Bl is N-R and B2 is C=O.
Intermediate amide, following protection of the D
functionality, is treated with a strong base such as KOtBu or KH and an alkylating agent such as methyliodide or dimethylsulfate to provide the intermediate where B1 is N-R.
This can be converted using the procedures outlined in Scheme I to afford the target compound.
WO 9~/2006~
2 PCT/US9`1/02283 Scheme XXI ~ 9 HD K N~ 1) Protection HD K N~
\)~( o 2) strongbase, \~4 0 ~ A/~ alkylating agent as described target where R = lower alkyl in Scheme I
Scheme XXII describes the synthesis of compounds of the invention in which group G is substituted with a tetrazole ring. Keto aldehyde (prepared as described in Scheme IX F) is treated with hydroxylamine hydrochloride in dimethylformamide to provide the nitrile. Following deprotection to the acid, it is coupled to provide target compounds wherein G is substituted with a nitrile group. Treatment with trimethylsilylazide and nBu2SnO followed by deprotection affords the target compounds.
WO 91/20062 PCT/US9~/02283 Scheme ~Xll 94 ~
1 HONH2-HCl ~9/l----¢~/
~CHO 0~CO2R CN /J~CO2R
t~D K B ,B2Z CN 5~ ~
~4 X~ D K B ,B2Z
n(~ )~ )m n(~)m ) TMSN3 Y ll x' nBu2SnO ~/~
2) Deprotect ~
D K B~B2Z
n(~A~ )m WO9~/2006' 21~ 7 ~12 PCT/US9~/02283 Scheme XXIII provides a synthetic scheme for the preparation of compounds where D is N-R. Amine intermediate (prepared as described in Scheme III) is converted to the trifluoroacetamide. Treatment with a strong base, such as KOtBu and an alkylating agent such as methyl iodide or dimethylsulfate followed by cleavage of the trifluorocetamide provides the intermediate amine wherein D is N-R. This is converted to compounds of the invention using procedures outlined in Scheme III.
WO 9~/20062 n~ PCT/US9~1/02283 Scheme XXIII - ~u- -Z157412 o 1~
\ HN Bl-B2--Z 1) KOtBu n(~ )m 1 ) TFA, CDI )_~ (CH3)2S04 g\ A/~ 2) KOH, CH30H
~ CH3 as described , Targets where D = -NCH3 n(~ ~A/Q)m in Scheme I
Persons of ordinary skill in the art will recognize that the foregoing schemes are exemplary only and should not be construed to be limiting.
Pharmaceutical preparations incorporating compounds according to the present invention can be used to block PKC
activity related to abnormal or undesirable cellular events and activity including tumorogeneis and cellular activity related to inflammation and reperfusion injury. Treatment of disorders and disease conditions can be performed by administration of effective amounts of pharmaceutical preparation that comprise compounds according to the present invention. Compounds can be formulated for human and animal prophylactic and therapeutic applications by those having ordinary skill in the art. The range of amounts of a compound to be administered to mammals, lS particularly humans, to be effective in inflammatory, tumor or reperfusion injury therapy can routinely be determined by those having ordinary skill in the art.
The compounds and pharmaceutical compositions of the invention may be administered by any method that produces contact of the active ingredient with the agent's site of action in the body of a mammal or in a body fluid or tissue.
These methods include but not limited to oral, topical, hypodermal, intravenous, intramuscular and intraparenteral methods of administration. The compounds may be administered singly or in combination with other compounds of the invention, other pharmaceutical compounds such as chemotherapeutic compounds, or in conjunction with therapies such as radiation treatment. The compounds of the invention are preferably administered with a pharmaceutically acceptable carrier selected on the basis of the selected route of administration and standard pharmaceutical practice.
The compounds of the invention are administered to mammals, preferably humans, in therapeutically effective amounts which are effective to inhibit protein kinase C, to inhibit tumor cell growth, inhibit inflammation of tissue, inhibit keratinocyte cell proliferation, inhibit oxidative burst from neutrophils or inhibit platelet aggregation. The W094/2006' PCT/US94/02283 2157~1~ I 98 -dosage administered in any particular instance will depend upon factors such as the pharmacodynamic characteristics of the compound of the invention, its mode and route of adminis-tration; age, health, and weight of the recipient; nature and extent of symptoms; kind of concurrent treatment, frequency of treatment, and the effect desired.
Compounds according to the present invention inhibit the activity of PKC in cells. The range of the amount of inhibitory compound that is effective for inhibiting PKC
activity can be determined by one having ordinary skill in the art. By inhibiting PKC activity, balanoids are useful in the treatment of disease conditions in which control of cellular growth, regulation and/or differentiation is desirable. An effective amount of a balanoid can be administered to mammals who are suffering from inflammatory, cardiovascular or neoplastic diseases, particularly inflammation, reperfusion injury and cancer, in order to counter the disease at the cellular level.
It is contemplated that the daily dosage of a compound of the invention will be in the range of from about 1 ~g to about lOO mg per kg of body weight, preferably from about 1 ~g to about 40 mg per kg body weight, more preferably from about lO ~g to about 20 mg per kg per day.
Pharmaceutical compositions of the invention may be administered in a single dosage, divided dosages or in sustained release forms. Persons of ordinary skill will be able to determine dosage forms and amounts with only routine experimentation based upon the considerations of this inven-tion. Isomers of the compounds and pharmaceutical compositions, particularly optically active stereoisomers, are also within the scope of the present invention.
The compounds of the invention may be administered as a pharmaceutical composition orally in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions. The compounds may also be administered parenterally in sterile liquid dosage forms or topically in a carrier. The compounds of the ~094/20062 2 1 5 7 ~ 1 2 PCT/US94/02283 _ 99 _ invention may be formulated into dosage forms according to standard practices in the field of pharmaceutical preparations.
See ~emington's Pharmaceutical Sciences, A. Osol, Mack Publishing Company, Easton, Pennsylvania.
Compounds of the invention may be mixed with powdered carriers, such as lactose, sucrose, mannitol, starch, cellulose derivatives, magnesium stearate, and stearic acid for insertion into gelatin capsules, or for forming into tablets. Both tablets and capsules may be manufactured as sustained release products for continuous release of medication over a period of hours. Compressed tablets can be sugar or film coated to mask any unpleasant taste and protect the tablet from the atmosphere or enteric coated for selective disintegration in the gastrointestinal tract.
Liquid dosage forms for oral administration may contain coloring and flavoring to increase patient acceptance, in addition to a pharmaceutically acceptable diluent such as water, buffer or saline solution.
For parenteral administration, a compound of the invention may be mixed with a suitable carrier or diluent such as water, an oil, saline solution, aqueous dextrose (glucose) and related sugar solutions, glycols such as propylene glycol or polyethylene glycols. Solutions for parenteral administration contain preferably a water soluble salt of the compound of the invention. Stabilizing agents, antioxidizing agents and preservatives may also be added. Suitable antioxidizing agents include sodium bisulfite, sodium sulfite, and ascorbic acid, citric acid and its salts, and sodium EDTA.
Suitable preservatives include benzalkonium chloride, methyl-or propyl-paraben, and chlorbutanol.
Animal studies have shown that perhaps 50% or more of ischemic-related myocardial damage can be attributed to polymorphonuclear leukocytes (neutrophils) which accumulate at the site of occlusion. Damage from the accumulated neutrophils may be due to the release of proteolytic enzymes from the activated neutrophils or the release of reactive oxygen intermediates (ROI). Much of the "no reflow" phenomenon 2,15~12 - 100 -associated with myocardial ischemia is attributed to myocardial capillary plugging. The plugging of capillaries has been attributed to both aggregated platelets and aggregated neutrophils. Although both cell types are aggregate during the ischemic event, the relative contribution of each to capillary plugging has not yet been established. It is accepted that the damage by neutrophils to myocardial tissue proceeds through a cascade of events, one of the earliest being the bonding of activated neutrophils to damaged vascular endothelium. However the binding of the neutrophils is significantly enhanced by their activation. Thus, an even earlier event is the generation of molecules (such as cytokines, and chemotactic factors) which can function as activation stimuli. These molecules probably originate from damaged and aggregated platelets, from damaged vascular endothelium, or from the oxidation of plasma proteins or lipids by endothelial-derived oxidants.
Strategies for overcoming the deleterious effects of reactive oxygen intermediates have centered on the development of scavengers for the molecules. Superoxide dismutase (SOD) has been shown to be a particularly effective scavenger of superoxide, but suffers from a very short half-life in the blood. Several companies have approached this problem by creating versions of the enzyme with increased half-lives by techniques such as liposome encapsulation or polyethylene glycol conjugation. Reports on the effectiveness of these new versions are mixed. Catalase, a scavenger of hydrogen peroxide, and hydroxyl radical scavengers have also been tested and found to be effective to varying degrees. However, none of the strategies designed to scavenge reactive oxygen intermediates will prevent the aggregation of platelets, the release of chemotactic molecules, the activation and adherence of neutrophils to vascular endothelium, or the release of proteolytic enzymes from activated neutrophils.
One advantage of protein kinase C inhibitors as therapeutics for reperfusion injury is that they have been demonstrated to: 1) block platelet aggregation and release of W094l20062 21~ 7 ~12 PCT/US94102283 neutrophil activating agents such as PAF; 2) block neutrophil activation, chemotactic migration, and adherence to activated or damaged endothelium; and 3) block neutrophil release of proteolytic enzymes and reactive oxygen intermediates. Thus, these agents have the capability of blocking all three of the most significant mechanisms of pathogenesis associated with reperfusion injury and should thus have a decided therapeutic advantage.
The table below contains a list of compounds according to the present invention. For convenience, in some cases the substituient groups described in their noun form rather than as adjectives, e.g. pyridine rather than pyridyl. It is to be understood that, unless specified, points of attachment of these functional groups can be any which are typically found in organic chemistry. Thus, for example, recitation of "pyridine"
as a substituient comprehends attachment at the 2, 3, or 4 position .
Cmpd. A 81 ~ 2 Z D E F G X K m n NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3 phenyl 2 NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3 phenyl 3 NH NH SO2 p-methyl phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3 phenyl 4 NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO phenyl CO H 1 3 phenyl CH2 NH CO p-hydroxy phenyl 0 2,6-dihydroxy CH2 phenyl CO H 1 3 phenyl 6 CH2 NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3 phenyl 7 CH2 NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-carboxy phenyl CO H 1 `3 phenyl 8 NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-carboxy phenyl CO H 1 3 phenyl 9 NH NH CO pyridine 0 3,5-dihydroxy CO 2-- I,UAy 6- CO H 1 3 phenyl hydroxy benzene NH NH CO pyrrole 0 3,5-dihydroxy CO 2-~.~ b~)A~6- CO H 1 3 phenyl hydroxy phenyl 11 NH NH CO oxazole 0 3,5~" 'hoA~ CO 2-- bUAY 6- CO H 1 3 phenyl methyl phenyl 12 NH NH CO indole 0 3-hydroxy-5- CO 2-hydroxy-6- CO H 1 3 r `h~l~ h_.,~1 methylphenyl 13 NH NH CO purine 0 3 1 `h y 5- CO 2,6-dimethyl CO H 1 3 i~
hJII h~ l phenyl Cmpd. A 8 1 ~ 2 ¦ D E F ¦ G ¦ X K ¦ m n 14 NH NH CO ~uran O 3-hydroxy CO 2,6-~tn hvAy CO H 1 3 phenyi phenyi NH NH CO lI,k~i-,_.,e O 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3 phenyi phenyi 16 NH NH CO PJ-i ' ' ,e O 3,5~1n 'hGA~ CO 2~ L.oAf 6- CO H 1 3 phenyl hydroxy benzene 17 NH NH CO ,~".i ,hli.,e O 3-hydroxy-5- CO 2~arboxy4- CO H 1 3 . . ._lhJ4 1 ~i hydroxy benzene 18 NH NH CO pyrazine O 3.5~' '~ Y CO 2~ IJVA~ 6- CO H 1 3 phenyi hydroxy benzene 19 NH NH CO imidazole O 3-hydroxy-5- CO 2~ LVA~r 6- CO H 1 3 ~
... `hy4h_.,~1 hydroybenzene ~ i--NH NH CO Ihiazole O 3,5-dihyroxy CO 2- I,oAf 6- CO H 1 3 ~ CJ~
benzene hydroxy benzene w 21 NH NH CO isoxazole O 3,5-d' . IhuAy CO 2~ boAy 6- CO H 1 3 ~ i~
phenyi hydroxy benzene l~
22 NH NH CO pyrazole O 3,5~dihydroxy CO 2- bvAy 6- CO H 1 3 phenyi hydroxy benzene 23 NH NH CO i~ r~l',' 'e O 3,5-dihydroxy CO 2~- LUA~6- CO H 1 3 benzene hydroxy benzene 24 NH NH CO benzene O 3,5~" . hvAy CO 2~ bOAY 6- CO H 1 3 phenyi hydroxy phenyl NH NH CO methyl benzene O 3-hydroxy-5- CO 2,6-dimethyl CO H 1 3 ~?
,.. _lh,1~ h_.. ~1 phenyi r~
26 NH NH CO dimethyl benzene O 3,5~ ./ CO 2,6~di,.. _1hvAy CO H 1 3 0 phenyl phenyi i~-27 NH NH CO trimethyi benzene O 3,5-dihydroxy CO 2~arboxy-6- CO H 1 3 benzene hydroxy benzene ~.:~ o Cmpd. A dl ¦ ~ Z ¦ D E F ~I ¦ X K m ¦ n ~ æ
28 NH NH CO Mr~.. h~1 0 3-hydroxy-5- CO 2~arboxy~ CO H 1 3 hyl~,h~" /l hydroxy benzene 29 NH NH CO ethyl benzene 0 3,5-dihydroxy CO 2-carboxy~ CO H 1 3 benzene hydroxy phenyl NH NH CO tetraethyl benzene 0 3-hydroxy CO 2~ OAY 6- CO H 1 3 phenyl hydroxy benzene 31 NH NH CO propyl benzene 0 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3 phenyl phenyl 32 NH NH CO tetra propyl 0 3,5-dihydroxy CO 2-carboxy~ CO H 1 3 benzene benzene methyl phenyl 33 NH NH CO butyl benzene 0 3-hydroxy-5- CO 2-carboxy-6- CO H 1 3 methoxy phenyl hydroxy benzene 34 NH NH CO tetrabutyl benzene 0 3,5-dihydroxy CO 2~ I,oAt 6- CO H 1 3 benzene hydroxy benzene NH NH CO pentyl benzene 0 3,5-dihydroxy CO 2-carboxy-6- CO H 1 3 benzene hydroxy benzene 36 NH NH CO t~ benzene 0 3,5-dihydroxy CO 2,6t'. hùAy CO H 1 3 phenyl phenyl 37 NH NH CO methoxy benzene 0 3,5-dihydroxy CO 2,6-dimethyl CO H 1 3 benzene phenyl 38 NH NH CO di~ hUAy benzene 0 3,5~i,.. 4hUAt CO 2~ LOAY 6- CO H 1 3 phenyl methylphenyl 39 NH NH CO td~ thoAy benzene 0 3,5-dlhydroxy CO 2~ I~OAY 6- CO H 1 3 benzene methoxy phenyl u NH NH CO tetra methyl 0 3-hydroxy-5- CO 2-carboxy~- CO H 1 3 ~, benzene methoxy phenyl hydroxy phenyl x~
41 NH NH CO ethoxy benzene 0 3,5-dihydroxy CO 2; t~oAy 6- CO H 1 3 benzene hydroxy phenyl Cmpd ¦ A E1¦ d l Z ¦ D E F G X K m n 42 NH NH CO dlethoAy benzene 0 3-hydroAy-S- CO 2-- boA~ 6- CO H 1 3 methyl phenyi methoxy phenyi 43 NH NH CO nitro benzene 0 3,5-dihydroAy CO 2,6-dimethyl CO H 1 3 benzene phenyl 44 NH NH CO dinitro benzene 0 31 ~h y 5- CO 2~ bUAY 6- CO H 1 3 methyi phenyi hydroxy benzene NH NH CO halo benzene 0 3,5-dihydroAy CO 2~arboAy4- CO H 1 3 benzene hydroxy phenyi 46 NH NH CO dihalo benzene 0 3-hydroxy CO 2-carboxy4- CO H 1 3 phenyl hydroxy phenyi 47 NH NH CO trihalo benzene 0 3,5-dihydroxy CO 2-carboxy4- CO H 1 3 phenyl hydroxy phenyi I 1 4~ NH NH CO tetrahalo benzene 0 3,5-dihydroxy CO 2~ bUA~ 6- CO H 1 3 benzene methoxy phenyl --49 NH NH CO benzene_ bUA~1 0 3,5-~L. huAy CO 2-carboxy4- CO H 1 3 1 acid phenyi methoxy phenyi NH NH CO benzene 0 3,5-dihydroxy CO 2~arhoAy4- CO H 1 3 d;~IIIUAYI;~ acid benzene methyi phenyi 51 NH NH CO b~" .id~ amido 0 3 m 'h /5- CO 2-hydroxy4- CO H 1 3 benzene methyi phenyl methyi phenyi 52 NH NH CO benzene dbmide 0 3-hydroxy CO 2,6-dimethyi CO H 1 3 phenyi phenyi 53 NH NH CO phenol 0 3,5-dihydroxy CO 2,6-d;,,,~lhuAy CO H 1 3 c benzene phenyi u 54 NH NH CO dihydroxy benzene 0 3,5-dihydroAy CO 2,6~ihydroxy CO H 1 3 phenyi phenyl NH NH CO trihydroxy benzene 0 3,5-dihydroxy CO 2-carboxy4- CO H 1 3 benzene methoxy phenyi Cmpd. A dl E2 ¦ Z D ¦ E F G X K m n l~
56 NH NH CO pe... 'hJd~vA~ O 3hJd~uA~t5- CO 2~ LUA~6- CO H 1 3 benzene methyi phenyi methoxy phenyi 57 NH NH CO triethoAy benzene 0 3,5-dlhydroxy CO 2,6-dimethyi CO H 1 3 benzene phenyi 58 NH NH CO tetra ethoxy O 3-hydroxy CO 2,6-dihydroxy CO H 1 3 benzene phenyi phenyi S9 NH NH CO propoxy benzene 0 3,5-dihydroxy CO 2 carboxy~ CO H 1 3 benzene hydroxy phenyi NH NH CO ~ lU~UAy benzene 0 3n.- .y5- CO 2~arboAy~ CO H 1 3 methyi phenyi hydroxy phenyi 61 NH NH CO t~i~,u~oAybenzene O 3,5-dihydroxy CO 2-t~rboxy-6- CO H 1 3 benzene methyi phenyi 62 NH NH CO tetra propoAy O 3,5-dihydroxy CO 2,6-" hùA~t CO H 1 3 benzene phenyi phenyi tJ
63 NH NH CO amidine 0 3,5-dihydroAy CO 2 carboxy~ CO H 1 3 phenyi hydroxy phenyi 64 NH NH CO dbmino benzene 0 3,5-dihydroxy CO 2-1,/.' UAY 6- CO H 1 3 benzene methyi phenyi NH NH CO methoxy pyridine 0 3,5-dihydroxy CO 2-o bVA'Y 6- CO H 1 3 benzene methyi phenyi 66 NH NH CO di,.. _lhuAy pyndine 0 3-hydroxy-5- CO 2-n jJUXY 6- CO H 1 3 methoxy phenyi methoxy phenyi 67 NH NH CO hydroxy pyridine 0 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3 c benzene phenyi v 68 NH NH CO dihydroxypyridine O 3,5-dihydroxy CO 2,6t-i.. ,~ll.GAy CO H 1 3 benzene phenyi 69 NH NH CO ethoAy pyrrole 0 3,5-dihydroxy CO 2-hydroxy-6- CO H 1 3 phenyi methyi phenyi ¦Cmpd. A El ¦ E2 D E F G X ¦ K m n NH NH CO dihydroxy pyrrole 0 3,5-dihydroAy CO 2-carboAy4- CO H 1 3 benzene hydroAy phenyl 71 NH NH CO dim h~JAy Indole 0 3-hydroAy-S- CO 2~arboAy4- CO H 1 3 methoxy phenyl hydroxy phenyl 72 NH NH CO hydroAy purine 0 3,5-dihydroAy CO 2-. bUA~ 6- CO H 1 3 benzene me~hyl phenyl 73 NH NH CO ~ hUAy ~uran 0 3: ~h J5- CO 2~ bUAy 6- CO H 1 3 methyl phenyl me~hoAy phenyl 74 NH NH CO hydro)yll,k~h~.,e 0 3,5-dihydroAy CO 2,6-dime~hyl CO H 1 3 benzene phenyl NH NH CO melhoAy p~ e 0 3,5-dihydroAy CO 2.6-dihydroxy CO H 1 3 benzene phenyl 76 NH NH CO 'i~.. lhùAy O 3-hydroAy CO 2,6-~- hùAy CO H 1 3 ~ r~
~".iJ~;"e phenyl phe~nyl ~1 77 NH NH CO hydroxy~,.;'".e O 3,5-dihydroAy CO 2-carboAy4- CO H 1 3 1 -~
phenyl hydroAy phenyl 78 NH NH CO diamido pf~ . e O 3,5-dihydroAy CO 2-carboxy4- CO H 1 3 J~
benzene hydroxy phenyl 79 NH NH CO amido pyrazine 0 3,5-'.lhuA1 CO 2~ bùAt 6- CO H 1 3 phenyl methoAy phenyl NH NH CO dielho)ty pyrazine 0 3 5-dihydroAy CO 2-carboAy4- CO H 1 3 benzene hydroxy phenyl 81 NH NH CO p-hydroAy phenyl O pyndine CO 2-~rboAy4- CO H 1 3 c melhyl phenyl v 82 NH NH CO p-methoAy phenyl O pyrrole CO 2-hydroAy4- CO H 1 3 me~hyl phenyl 83 NH NH CO p-chloro phenyl O oxazole CO 2,6-~" huAy CO H 1 3 phenyl Cmpd. A E~ Ez Z D E F G X K m n ~, 84 NH NH CO~ hUA~ phenyi O indole CO 2,6-dihydroxy CO H 1 3 phenyi NH NH CO .. hJd~UA~ phenyi O purine CO 2- buAf 6- CO H 1 3 hydroxy phenyi 86 NH NH COm-chloro phenyi O furan CO 2-. buA~6- CO H 1 3 hydroxy phenyi 87 NH NH CO o-methoxy phenyi O Ihlu~ .. e CO 2-hydroxy4- CO H 1 3 methyi phenyi 88 NH NH CO o-hydroxy phenyi- O ~,.; ' ,e CO 2,6-dimethyi CO H 1 3 phenyi 89 NH NH CO o-chloro phenyi O ~".i ' .e CO 2,6-. . hùAy CO H 1 3 phenyi NH NH CO p-hydroxy phenyi O pyrazine CO 2-carboxy4- CO H 1 3 o methoxy phenyi I
91 NH NH CO m-methoAy phenyi O imidazole CO phenyi CO H 1 3 92 NH NH CO m-hydroxy phenyi O Ihiazole CO 2-carboxy4- CO H 1 3 methyi phenyi 93 NH NH CO p-methoAy phenyi O Isoxazole CO 2-carboxy4- CO H 1 3 hydroxy phenyi 94 NH NH CO o-methoAy phenyi O pyrazole CO 2-carboxy4- CO H 1 3 hydroxy phenyi NH NH CO m-hydroA~ phenyi O Is~tl,' 'e CO 2-carboxy4- CO H 1 3 hydroxy phenyi u 96 NH NH CO o-methoAy phenyi O benzene CO 2-carboxy4- CO H 1 3 o methyi benzene 97 NH NH CO o-chloro phenyi O methyibenzene CO 2-carboxy4- CO H 1 3 methyi benzene CmDd. A E1 E2 Z D E F ~ X ¦ K m n 98 NH NH C0 p-hydroxy phenyl O dimethyl CO 2~ bUA'y6- CO H 1 3 benzene hydroxy phenyl 99 NH NH C0 p-chloro phenyt O trlmethyl CO 2-carboxy-6- CO H 1 3 benzene me~hyl phenyl 100 NH NH C0 m-hydroxy phenyl 0 t- ... lh~ CO 2-l,JJ~uA~6- CO H 1 3 methyl phenyl 101 NH NH C0 ... m lhuAy phenyl O ethylbenzene CO 2,6-Ji~ hUAy CO H 1 3 phenyl 102 NH NH C0 p-hydroAy phenyl O tetraethyl CO 2-carboxy-6- CO H 1 3 benzene methoxy phenyl 103 NH NH C0 p-methoAy phenyl O propylbenzene CO 2.6-dihydroxy CO H 1 3 benzene phenyl 104 NH NH C0 m-hydroxy phenyl O tetrapropyl CO 2-hJJ~UAy6- CO H 1 3 0 ~-~
benzene methyl phenyl ~ ~n 105 NH NH C0 o-hydroxy phenyl O butyl benzene CO 2,6-dihydroxy CO H 1 3 phenyl 106 NH NH C0 m-hydroxy phenyl O tetrabutyl CO 2-carboxy-6- CO H 1 3 l~a benzene me~hyl benzene 107 NH NH C0 m-hydroxy phenyl O pentytbenzene CO 2-carboAy-6- CO H 1 3 methyl benzene 108 NH NH C0 o-hydroxy phenyl O t , ,t~ CO 2~rboxy-6- CO H 1 3 benzene hydroxy phenyl 109 NH NH C0 o-chloro phenyl O methoxy CO 2-carboAy-6- CO H 1 3 c benzene methoxy phenyl ~O
110 NH NH C0 p-methoxy phenyl O :' huAy CO 2-hydroxy-6- CO H 1 3 jR
benzene methyl phenyl . ~
Cmpd. ¦ A E~ E~ D E F G X K m n ¦ ~ ~ g 111 NH NH CO p-chloro phenyi O t~lr.. hoAy CO 2,6-dimethyi CO H 1 3 benzene phenyi 112 NH NH CO p-hydroxy phenyi O tetramethyi CO 2,6-dihydroxy CO H 1 3 benzene phenyi 113 NH NH CO p-rnethoxy phenyi O ethoxybenzene CO 2-carboxy4- CO H 1 3 methyi benzene 114 NH NH CO p-hydroxy phenyi O diethoxy CO 2-carboxy4- CO H 1 3 benzene hydroxy phenyi 115 NH NH CO m-hydroxy phenyi O nitrobenzene CO 2-carboxy4- CO H 1 3 methoxy phenyl 116 NH NH CO p-chloro phenyi O dinitrobenzene CO 2,6-dimethyi CO H 1 3 phenyi 117 NH NH CO o-methoxy phenyi O halobenzene CO 2-hydroxy4- CO H 1 3 1.
methyi phenyi O
118 NH NH CO p-methoxy phenyi O dihalobenzene CO 2-carboxy4- CO H 1 3 methyi phenyi 119 NH NH CO p-hydroxy phenyi O trihalobenzene CO 2-hydroxy4- CO H 1 3 methyi phenyi 120 NH NH CO o-chloro phenyi O tetrahalo CO 2-carboxy4- CO H 1 3 benzene methoxy phenyi 121 NH NH CO m-hydroxy phenyi O carboxyacid CO 2-carboxy4- CO H 1 3 benzene hydroxy phenyi 122 NH NH CO m-chloro phenyi O ' bOAj ld CO 2-carboxy4- CO H 1 3 c benzene ethyi benzene v 123 NH NH CO p-hydroxy phenyi O amido benzene CO 2,6-di".~t'loiy CO H 1 3 ,~
phenyi 00 124 NH NH CO o-methoxy phenyi O diamido CO 2-c~rboxy4- CO H 1 3 benzene hydroxy phenyi -Cmpd. A¦ B1 ¦ B2 ¦ Z D E F G ¦ X K m n ¦ o 125 NH NH CO p-hydroxy phenyi O phenol CO 2-carboxy4- CO H 1 3 hydroxy phenyl 126 NH NH CO 1.. hùAy phenyl O dihydroxy CO 2-; LUAY6- CO H 1 3 benzene hydroAvy phenyi 127 NH NH CO p-methoAy phenyi O trihydroAy CO 2-carboxy4- CO H 1 3 benzene propoxy benzene 128 NH NH CO 1.. 3~ UA~ phenyl O l~,t~ hJJ~UAy CO 2carboxy4- CO H 1 3 benzene hydroxy phenyi 129 NH NH CO p-chloro phenyi O triethoxy CO 2,6-dimethyi CO H 1 3 benzene phenyi 130 NH NH CO p-methoAy phenyi O tetraethoxy CO 2-carboxy4- CO H 1 3 benzene methoxy phenyi I
131 NH NH CO p-hydroxy phenyi O pentoAy CO 2,6-dihydroxy CO H 1 3 1~ C r~
benzene phenyl ~ _~
132 NH NH CO p-hydroxy phenyi O dipentoxy CO 2,6~ . t~10AY CO H 1 3 benzene phenyl 133 NH NH CO m-methoAy phenyl O tripentoAvy CO 2- buAy6- CO H 1 3 benzene ethyi benzene 134 NH NH CO o-hydroxy phenyi O I ~r~ Y CO 2-carboxy4- CO H 1 3 benzene . hydroxy phenyi 135 NH NH CO o-hydroxy phenyi O aniline CO 2-carboxy4- CO H 1 3 hydroxy phenyi 136 NH NH CO o-chloro phenyi O dbmino CO 2,6-~ thoAy CO H 1 3 C
benzene phenyi ~, 137 NH NH CO p-methoAy phenyi O methoxy CO 2-carboxy4- CO H 1 3 pyridine propyi benzene A
2157~
E
_ y I
O O O O O O O O O O O O O
N E N N N E N ~ N N N E N N ~D N _ N E N
O O O O O O O O O O O O O
~ C ~ t ~ 2 E
C~
O O O O O O O O O O O O O
~ ~ C G C ~ lii c ~ CL ~ t~. G
G G ~ O O O
m O O O O O O O O O O O O O
m Z Z Z Z Z Z Z Z Z Z Z Z Z
I
Z Z Z Z Z Z Z Z Z Z Z Z Z
c E 0 0 -- N ~ ~ It~ O
e~ e~ e~ e~ e!~ e~ e!~ e~ e~ eq e!~ e~ e~
E _ _ _ _ _ _ _ _ _ _ _ _ _ Y
- O O O O O O O O O O O O O
~ '~
e~ e~
'. '. eD~U ~U 'U
N . N . G C~ O .E ~ 2 : L
O O O O O O O O O O O O O
-2 2 3~ 3O~ ~ 3O~
LU ,. eD ~ g~ U g, ~ U ~ l U ~ I
e~ Ce!~ _ e!~ _ e!~ _er~ _ e!~ . eq . e~ _ e!~ . eq . e!~ _ eq .
O O OO O O O O OO O O O
~U ~ eD eu eu eeu ~ eu e ,eu eu eu O OG CL ~ O C~
m O OO O O O O OO O O O O
~ o m Z Z Zz z z z z zZ Z z z I
Z Z Zz z z z Z Zz Z z z c u~ eD r~ e~ u~~OD ~ ~ eD
C~ ~ _ __ _ _ _ _ __ _ _ _ _ Cmpd. ¦ A B1 B2 Z D E F G X K m n ¦ O
166 NH NH CO p4hloro phenyl 0 3,5-dihydroxy CO thbzole CO H 1 3 benzene 167 NH NH CO m-methoAy phenyl 0 3,5-dihydroxy CO Isoxazole CO H 1 3 benzene 168 NH NH CO p-chloro phenyl 0 3,5-dihydroxy CO pyrazole CO H 1 3 benzene 169 NH NH CO p-hydroxy phenyl O benzene isutl,' '~ CO H 1 3 170 NH NH CO p-methoAy phenyl 0 3,5-dihydroxy CO benzene CO H 1 3 benzene 171 NH NH CO o-hydroxy phenyi 0 3,5-dihydroxy CO methyibenzene CO H 1 3 benzene 172 NH NH CO o-chloro phenyl 0 3,5-dihydroxy CO dimethytbenzene CO H 1 3 ~' benzene ,~
173 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO trimethylbenzene CO H 1 3 benzene 174 NH NH CO p-me~hoxy phenyi 0 3,5-dihydroxy CO t~ tl,~l CO H 1 3 benzene 175 NH NH CO ,.. n lhoAy phenyl 0 3,5~ihydroxy CO ethylbenzene CO H 1 3 benzene 176 NH NH CO p-hydroxy phenyl 0 3,5-dihydro~ CO tetraethylbenzene CO H 1 3 benzene 177 NH NH CO m-hydroxy phenyt 0 3,5-dihydroxy CO propyibenzene CO H 1 3 c benzene benzene u 173 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO tetrapropyi CO H 1 3 benzene benzene 179 NH NH CO p-chloro phenyl 0 3,5-dihydroxy CO butylbenzene CO H 1 3 _ Y
- O O O O O O O O O O O O O
~ "
~ n ~ ~. .. ~
~ , . C . ~ . ~ _ o o o o o o o o o o o o o o J~ ' ; ~ ~L ~ o ~ O ~ o2 2 o o o o o o o o o o o o o ,c C c~ C~ C C~ c 1~ ~ Q, G 'c~ ~
c~ ~ G ~ c~ ~. G ~ G
,o ,o ,o C- ~ O O G CL G i o m I
E 0 0 '`~ D ~ 0 ~ o, Cmpd. A ¦ B~ B2 Z D E F G X K m n ~ 2 193 NH NH CO ... h~J~UAy phenyi 0 3,5-dihydroAy CO trihalobenzene CO H 1 3 benzene 194 NH NH CO m-chloro phenyi 0 3,5-dihydroAy CO tetrahalobenzene CO H 1 3 benzene 195 NH NH CO p-hydroxy phenyi 0 3,5"hyJ,JA~ CO carboxyadd CO H 1 3 benzene benzene 196 NH NH CO p-chloro phenyi 0 3,5-dihydroAy CO " ~ ~,f ld CO H 1 3 benzene benzene 197 NH NH CO p-methoAy phenyi 0 3,5-dihydroxy CO amido benzene CO H 1 3 benzene 198 NH NH CO m-hydroA~y phenyi 0 3,5-dihydroAy CO diamido benzene CO H 1 3 benzene 199 NH NH CO m-chloro phenyi 0 3,5-dihydroxy CO 3,5-dihydroxy CO H 1 3 1' benzene benzene a~
200 NH NH CO o-methoAy phenyi 0 3,5-dihydroA~y CO trihydroxybenzene CO H 1 3 benzene 202 NH NH CO o-methoAy phenyi 0 3,5-dihydroA~y CO t `r~hJJ~uAy CO H 1 3 benzene benzene 202 NH NH CO p-chloro phenyi 0 3,5-dihydroAy CO p~.~' hJJ~uA~ CO H 1 3 benzene benzene 203 NH NH CO p-chloro phenyi 0 3,5-dihydroAy CO triethoAybenzene CO H 1 3 benzene 204 NH NH CO o-hydroAy phenyi 0 3,5-dihydroxy CO tetraethoAy CO H 1 3 c benzene benzene r~
205 NH NH CO p-hydroAy phenyi 0 3,5-dihydroAy CO pentoxybenzene CO H 1 3 benzene ~, 206 NH NH CO p-hydroxy phenyi 0 3,5-dihydroxy CO dipentoxybenzene CO H 1 3 benzene Cmpd. A B~ ¦ J2 ¦ ¦ E F G X K m n 207 NH NH CO p-methoAy phenyl 0 3,5~ihydroxy CO tripen~oxybenzene CO H 1 3 benzene 208 NH NH CO m-hydroxy phenyl 0 3,5-dihydroxy CO l~t,., , y CO H 1 3 benzene benzene 209 NH NH CO o-methoAy phenyi 0 3,5-dihydroxy CO amino benzene CO H 1 3 benzene 210 NH NH CO o-chloro phenyi 0 3,5-dihydroxy CO diamino benzene CO H 1 3 benzene 211 NH NH CO p-methoxy phenyi 0 3,5-dihydroxy CO methoxypyridine CO H 1 3 benzene 212 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO Ji",_lhuA~pyridine CO H 1 3 benzene I ~V
213 NH NH CO p-methoxy phenyi 0 3,5-dihydroxy CO hydroAypyridine CO H 1 3 benzene 214 NH NH CO p-methoxy phenyl 0 3,5-dihydroxy CO dihydroxypyridine CO H 1 3 1 ~_~
benzene l~
215 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO ethoAypyrrole CO H 1 3 benzene 216 NH NH CO ... hJJ~uAy phenyi 0 3,5-dihydroxy CO dihydroxypyrrole CO H 1 3 benzene 217 NH NH CO p-chloro phenyi 0 3,5-dihydroxy CO ' . `i~.. A~Hndole CO H 1 3 benzene 218 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO hydroAypurine CO H 1 3 c benzene u 219 NH NH CO p-methoxy phenyi 0 3,5-dihydroxy CO d~.,._lhuAyfuran CO H 1 3 benzene 220 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO hydroxyll, ~ .,t CO H 1 3 benzene ~ o Cmpd. A ~1¦ i32 Z D E F G X K m n Wg 221 NH NH C0 m-chloro phenyi O benzene CO methoAyl".; ' ,e CO H 1 3 222 NH NH C0 p-hydroAy phenyi 0 3,5-dihydroAy CO 'i.. IhuAy CO H 1 3 benzene IJ j . ;~;. ,e 223 NH NH C0 p-chloro phenyl O benzene CO hydroAyp,.; " ,e CO H 1 3 224 NH NH C0 m-chloro phenyi 0 3,5-dihydroAy CO diamido py.; .;di,.e CO H 1 3 benzene 225 NH NH C0 o-hydroAy phenyi 0 3,5-dihydroxy CO amido pyrazine CO H 1 3 benzene 226 NH NH C0 m-hydroAy phenyl 0 3,5-dihydroA~y CO diethoAypyrazine CO H 1 3 benzene 227 NH NH C0 quinoline 0 3,5-dihydroAy CO phenyi CO H 1 3 1--benzene 22a NH NH C0 methoxy 0 3,5-dihydroxy C0 phenyi CO H 1 3 quinoline benzene 229 NH NH C0 ~'. hùAy 0 3,5-dihydroAy C0 phen~ CO H 1 3 quinoline benzene 230 NH NH C0 I,i hUA~ O 3,5-dihydroAy CO phenyl CO H 1 3 quinoline benzene 231 NH NH C0 hydroAy 0 3,5-dihydroxy C0 phenyi CO H 1 3 quinoline benzene 232 NH NH C0 dihydroAy 0 3,5-dihydroAy C0 phenyl CO H 1 3 c quinoline benzene u 233 NH NH C0 ethoAy 0 3,5-dihydroxy C0 phenyl CO H 1 3 ~;
quinoline benzene 234 NH NH C0 amino 0 3,5-dihydroAy CO phen~ CO H 1 3 quinoline benzene Cmpd. A 31 B2 7 D E ¦ F G ¦ X ¦ K m n 235 NH NH CO dbmido 0 3,5-dihydroxy CO phenyi CO H 1 3 qulnoline benzene 236 NH NH CO trihalo 0 3,5-dihydroAy CO phenyi CO H 1 3 quinoline benzene 237 NH NH CO quinolinecarboA~yic O 3,5-dihydroAy CO phenyi CO H 1 3 acid benzene 238 NH NH CO ~,ln l.e O 3,5-dihydroAy CO phenyi CO H 1 3 benzene 239 NH NH CO methoAy O 3,5-dihydroAy CO phenyi CO H 1 3 ~il ' , " ,e benzene 240 NH NH CO . ~lh~AY O 3,5-dihydroAy CO phenyi CO H 1 3 'il' . ' ,e benzene 241 NH NH CO l~i\, Ihv~ O 3,5-dihydroAy CO phenyi CO H 1 3 ~ r~
il' ,- ',e benzene ~D
242 NH NH CO hydroxy O 3.5-dihydroAy CO phenyi CO H 1 3 1 _~
, ', ' ,e benzene ~p.
243 NH NH CO t~;hy.' VAY O 3,5-dihydroxy CO phenyi CO H 1 3 ~,-ln !ine benzene 244 NH NH CO t hv~y O 3,5-dihydroAy CO phenyi CO H 1 3 e benzene 245 NH NH CO diamino 0 3,5-dihydroxy CO phenyi CO H 1 3 il~ln ' ,e benzene 246 NH NH CO triamido 0 3,5-dihydroAy CO phenyi CO H 1 3 q~lr ' ,e benzene ~
247 NH NH CO tetrahalo 0 3,5-dihydroAy CO phenyi CO H 1 3 O
, ', 'i. ~e benzene 248 NH NH CO 'i"~ ~'h,e O 3,5-dihydroxy CO phenyi CO H 1 3 Ji~_. i VAyi- acid benzene 21S7~ 120-E _ _ _ _ _ _ _ _ _ _ _ _ _ _ O O O O O O O O O O O O O O
C C C C ~ &
G 1~ C~ ~ G C~ C~ ~ C~ 1~ D ~ C~ C~
LL
ID
o ~ ~ O ~
O O O O O O O O O O O O O O
C ~ ~ C C ~D ~ G C~ ~ ~ G
: : : . : :
G G t~ o o ; G ~ C t~
m O O O O O O O O O O O O O O
m Z Z Z Z Z Z Z Z Z Z Z Z Z Z
ZZZZZZZZZZZZZZ
-E ~ O -- N 0 ~ U) ~ ~-- 0 CO O _ N
~le N N N N N N N N N N N N N N
WO 94/20062 215 7 ~ 12 PCT/US94/02283 E _ _ _ _ _ _ _ _ _ _ _ _ _ Y
x 8 ~, ~, ~, ~, 8 ~, ~, ~ ~
G G C~ Cl. ~ G ~ CL ' ' C
O O O O O O O O O O O O O
o ' o ! ' 7` J ~ ~ _ r O O O o o o o o o o o o o -- C~' c ~ S c Q ~ G ~ G ~ ~, ~ ~, o ' o 1' o C C o Q. O O 11 t~ G O O O
N
m O O O O O O O O O O O O O
m Z Z Z Z Z Z Z Z Z Z Z Z Z
-T T I I I T I I ~
Z Z Z Z Z Z Z Z Z Z Z Z Z
~
C.) N N N N N NN N N ~N` N C~l N
WO 94/20062 PCTtUS94/02283 21~7412 - 122 -E
Y
O O O O O O O O O O O O O O
2 ~ o ~ o : : o ~ .E
8 8 8 ~, 8 8 8 8 8 8 8 8 8 8 2 ,~ D ~ ~ C ~ S
O O O O O O O O O O O O O O
G 0- G ~ ~ CL , ~ ~ ' ~ c G
c c~ G ~ c o o o `.
m O O O O O O O O O O O O O O
m ZzzzzzzzzzzzzZ
ZZZZZZZZZZZZZZ
E ~D ~ 0 0 c~ _ N C" ~ U~ ~ ~ 0 0 C~ ~1. N N N N N0 N0 N0 N N0 N0 N0 N N N0 WO 94120062 21~ 7 412 PCTIUS94/02283 E _ _ _ _ _ _ __ _ _ _ _ Y
~ ~ ,,. . ~
_ 8 8 8 o 8 8 8 8 8 8 8 8 8 o o o o o o o o o o o o o G ,c ~ ~ ~ G G ,c . . O : ' G ~ . C G O
m O O O O O O O O O O O O O
m I I I I I I I I I I
Z Z Z O O O ~ Z c~ ~o Z Z Z Z Z
~ ' N N N N 0 N 0 r-- 0 0 0-- N
cn ~`~
Cmpd. A B 1 B 2 Z D E F G X K m n f 303 NH O CO m-hydroxy phenyi O 3,Wihydroxy CO phenyi CO H 1 3 benzene 304 NH N-ethyi CO o-chloro phenyi O 3,5-dihydroAy CO phenyi CO H 1 3 benzene 305 NH N- CO ... mlhuAy phenyi O 3,5-dihydroxy CO phenyi CO H 1 3 methyi benzene 306 N-anisly N- CO p-hydroxy phenyi O 3,5-dihydroAy CO ;l~ln 'ine CO H 1 3 phenyi benzene " bGAJr': acid 307 NH NH CS p-chloro phenyi O 3,5 :"hJ ' UAy CO phenyi CO H 1 3 benzene 308 NH NH CO p-hydroxy phenyi CH2 3,5-dihydroAy CO phenyi CO H 1 3 benzene 309 NH NH CO p-hydroxy phenyi N-ethyi 3,5-dihydroAy CO phenyi CO H 1 3 N
310 NH NH CO p-methoxy phenyi N- 3,5-dihydroAy CO phenyi CO H 1 3 phenyi benzene 311 NH NH CO o-methoxy phenyi N- 3,5-dihydroxy CO phenyi CO H 1 3 propyi benzene 312 NH NH CO m-chloro phenyi O 3,5-dihydroxy CS phenyi CO H 1 3 benzene 313 NH NH CO m-chloro phenyi O 3,5-dihydroxy CH(OC phenyi CO H 1 3 benzene H ~ ~, 314 NH NH CO o-chloro phenyi O 3,5-dihydroAy CH(O- phenyi CO H 1 3 benzene phenyi) u 315 NH NH CO o-hydroxy phenyi O benzene CH2 phenyi CO H 1 3 O
316 NH NH CO m-chloro phenyi O 3,WihydroAy O phenyi CO H 1 3 benzene WO 94/20062 215 7 ~ 1~ PCT/US94/02283 r.~ -- r,~l ~ . _ _ N N r.~ r,~ _ _ N
-- _ _ _ N r~ ~ N e~ N
Y I I I I I T
8 8 8 8 8 8 8 8 8 8 8 8 ~ 8 ~
~ ;~, f ~ f f f f f ~ ~D f f ~~D
~ 8 8 8 8 8 8 8 8 8 8 ~ 8 ~ 8 ~i b ~ C C ~ D
~ . ~ . r, ~, r,.~, r, ~ rq _ ~ _ ~ . ~ . _ . C~ . .
O O O O O O O O O O O O O O O
c c ~ ~ ~ ~ ~ _ ~
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G G C~ i O C~ G 2 . : : .
_ 8 8 8 8 8 8 8 8 8 8 8 o 8 ~
m I I I I T
Z Z Z z z z z z z z a~ Z o. Z Z Z
E ~ N N CNU N N N N N N N c~ C~
2157 ~ 126 -N N _N _ 0 0 0 0 _ N 0 E __ _ _ _ _ _ _ _ _ _ _ N
Y II I I I I T I I I I T
Otn Ou~ O a~ O u~ Z, ,~ ~ O O
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fB o z 8 t~ 8 z E 8 z c~ ~, z 8 E . c OOOOOOOOOOzOOO
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m u~ O tn ~ ~ ~
m OO z ,~ ~ , ,~ . ~ I I
~ TT T T I C u~ ~ e ,-- T
E NC~ ~ u~ ` 0 O) O _ N 0 ~ It7 WO 94/20062 21~ ~ 112 PCT/US94/02283 --C~J N C'~ N ~ r~ _ Y
O O O O O o~ O a~ Ou~ O c/~ O a~
D ~ D 'D _ E _ 2 ~
~ . . . ~
0~ O~n O O O O U~ I _ I ~ O O O
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WO 94t20062 PCTtUS94/02283 2157ll~2 N ~t _ _ _ N N c~ _ _ N
E _ _ N ~ ~ N C'~ N
Y
O a~ O a~ Z~,~U O O o o cn O O O
'U
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m t~ O a~ z '' z E ' ~ ' ~
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c-- Co _ N 7 ~ In tD
WO 94/20062 215 7 ~12 PCT/US94102283 N ~ N ~ _ _ E _ _ _ _ _ _ _ _ _ _ _ N C'~
I T I I I I I T T T I I T
X N
O O O tn O o~ O a~ O o~ o 0, z C~ ~ ,'D ~ o ~ ; ~ ; ; ; , O O O O O cn O O a~ O
O Z G ~1) ~ 2 ~ 2 ~ o ' G ' Q ~ Q ~ ~ b . ~ . ~ . ~ . ~ . ~ ~ c -o o o o o o o o o o o o o G Q Q ~Q 'G C ~ D C ~ ~D
N ~ ~ Q ~ ~ ~, ,~
: . - . . : . ' --~ _ _ _ _ G O Q O O O G Q C
N
m O O O o O~n O O ~ O O O O
- N ~ ~ ~
I I I I IC~ O O Z 'Q Z E IZ Z ~CQ Z Q
N C,'7 111E ' Q ~ I I I I z E ~ o _ N C~
Cmpd. A ¦ 8 1 ¦ d 2 D E F (1 X K m n W
386 CH2 NH CS o-methoxy phenyi O methoxy CS amino benzene CS H 4 e 387 0 NH CO o-hydroxy phenyi O h 'hJAY CO diamino benzene CH(CH3 H 2 2 ~".; ' ' .e CH3~
388 S NH CO ... n, Iho,~y phenyi O hydroxy N- methoxypyridine CS H 3 2 .. IJi"e methyi 389 N-SO z NH CO p-chloro phenyi NH dbmido CO d,,,,~lhùxypyridine C=N- H 2 3 phenyi 1,,.; .. Ji"e phenyi 390 N-methy NH CO p-chloro phenyi O amido pyrazine N- hydroxypyridine CS H 1 3 pyridine 391 N-butyi NH CO p-hydroxy phenyi NH diethoxy CO dihydroxypyridine CO H 1 3 pyrazine 392 N-phenyi NH CO p-chloro phenyi NH phenyi N- phenyi CO H 1 3 1' purine o 393 S NH CO p-hydroxy phenyi NH phenyi CO phenyi CO H
394 N-SO z NH CS hydroxytl,', h~ne NH dihydroxy CH2 phenyi CO H
phenyi benzene 395 N-methyi NH CO methoxy~,.'' '.e CH2 dihydroxy CH2 phenyi CO H 1 4 benzene 396 N-butyl NH CS ~" . 'huAy O dihydroxy O phenyi CO H 2 ~,.;JaL;"e benzene 397 N-phenyi CH2 CO hydroxypf~ ' " ,e O dihydroxy O phenyi CO H 3 1 ~
benzene 6 398 N-pyridy O CS diamido ~".; ,;di"e N- dihydroxy S pheny CS H 4 1 phenyi benzene 399 0 0 CO amido pyrazine N- dihydroxy CH2 phenyi CO H 2 2 methyi benzene WO 94/20062 21~ 7 ~12 PCT/US94/02283 ._ N ~ ~r _ _ N _ N N _ N _ C'7 E ,., N
Y
8 8 8 ~ ~ 8 ~ 8 ' . , X~ ", X~ o I ~ ~, ;
D D ~D
_ _ -- -- C
8 8 8 8 t~ O G ~ 8 8 8 8 ~, o x x r O ~ r ~D ~ ~ -- S _ ~
Z D , '~ I I I I I N ~ _ '` 2 ~ ' ' ~ ~5, 'D o ~D D
~ . ~ . ~ _ ~ . Cl. ~ ~ G o m O O o m O o m O cn O m O o~
m , _ ' E I , rD .
~7 D E ,0 D E
c~ z z z z O o u~ Z ~ z z Z z cL
Eo _ N ~ ~ cn O-- N
~,~ ~ ~ Sj! ~ ~ ~ ~o ~ ~o ~
21S7~12 - 132 -~ ~ _ N ~t . _ _ N N ~ ~
E
N ~ ~ N C'~ N
o~ O ~ O cn O a~ O cn O O O c~ O
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m o ~ O m O m O m O O O O O O
m , .~ , ti; I , .~ , ,, I
T T T z ~ ~ o ~ z ~ z z z C
E' ~ ~ m ~ ~ 0 ~N N N ~ ~tN N
wo g4/20062 2 1 ~ 7 Ll 1 2 PCT/US94/02283 -N ~ N ~ _ _ _ N
E _ _ _ _ _ _ _ _ _ _ N C'~ ~t N
Y
~, 8 ~, 8 8 ~, 8 ~, 8 o 8 ~, 8 c~
8 z ~ 8 z ~ 8 o o o o 0 o 8 8 8 ' ' ~ ' ' E
Z ~ Z E z Z C z ~ I I I I
J
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Q . ~, ~ ~ , ' ; ; ; ~ _ ~ O ' ~ c e N
m 0 0 0 0 0 0 0 0 0 0 0 0 0 0 m T ~ O O Z ~ ' E I Z G Z ~ I
0 ,~ E o G ~
Z ~ Z Z Z Z O ~ Z Z Z Z (~ O 0 E r~ CD ~ ~ ~ N ~ ~ 10 PCTIUS94/On83 2157~ 134-N rr~ r.~ _ _ N _ _ N _ r.~7 rr, E ,~, N
Y I T I T I I I T T T T T
X I ~ --I U~ Z, ~,, o o o o o o ~D ~D
~ ~ G O G ~ C~ CL Q ~ C C~ Q
$ o c~ 8 ~ 8 u) 8 ~ o z ~ 8 J ,J . . ". ! rD !
o D D C r~ ' C
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o O ~ ' U ' 8 8 8 8 8 8 c~ 8 8 8 8 8 m I ~ r z s . D D Z S ' ~
<7 ~ E ~ ~ 1~ C 1 T T T T
E
Cmpd. A ¦ E1 ~2 Z D E F G ¦ X K m n F~
454 N-SO 2- NH CO p_., hy ' UAt NH benzene CS dihydroxy CS H 1 3 phenyi benzene benzene 455 N-methyi NH CO triethoxy benzene NH phenyi N- dihydroxy CO H 1 3 methyl benzene 456 N-but~1 NH CO tetraethoxy O phenyi N- dihydroxy CO H
benzene methyi benzene 457 N-phenyl NH CO pentoxy benzene O phenyl CO benzene CO H 1 2 458 N-pyridyi NH CS dipentoxy benzene O phenyi N-ethyi dihydroxy CO H 1 4 benzene 459 NH NH CO tripentoxy benzene O phenyi CO dihydroxy CS H 2 benzene 460 S NH CO l-,t~ O phenyi N- dihydroxy CO H 3 1 1' ~3 benzene purine benzene Ul t~
461 N-SO 2 NH CS aniline O phenyi CO dihydroxy CO H 4 1 1 ~
phenyi benzene ~"
462 N-methyi NH CO diamino benzene O phenyi CH2 dihydroxy CS H 2 2 benzene 463 N-buty NH CO methoxy pyridine O phenyi CH2 dihydroAy CO H 3 2 464 N-phenyi CH2 CO 'i,. `h ./ pyridine O phenyi O dihydroxy CO H 2 3 benzene 465 N-pyridyi O CO hydroxy pyridine O phenyi O dihydroxy CO H 1 3 benzene v 466 0 0 CS dihydroxy pyridine O phenyi S dihydroxy CO H 1 1 0 benzene 467 CH2 N- CO ethoxy pyrrole O phenyi CH2 dihydroxy CS H 1 3 phenyi benzene r~ _ N ~ _ _ _ N N e~
---- -- -- N ~ ~t N'~ N _ ~ _ Y
O O O O O O O a~ O O a~ O O
D
o ~ O
O O O U7 I c I Ç~ O O O O ~1~ 0 ~D ) tD ~D
D e~~D 'C~ ~D
u~ ! . ~ I ~ I : ~D 'C I,, I 'D ~CD ~. 'D
C ~C
O O O o o o o O O O O z ~ z E
'D 'D
:
D
N ,~ ~ . _ ~'D ~
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-m O O O O O o cn O ~ O C~ O O
Z E Z Z ~ Z ~ Z Z Z Z Z O O O Z c , C E D ~ ~ I
c E ~ 0 -- N ~ ~ O
WO 94/20062 215 ~ 412 PCT/US94/02283 N _ N N _ N _ ~ C'~ _ N ~
E _ _ _ _ _ _ _ _ _ _ _ _ _ _ Y
X Z ~ Z C
t~ 8 ~ 8 t~ 8 ~ 8 ~ O ~ c~
ID ID : I ! ! I ' rL
o E :
~ 8 ~ O Z G 8 ~, 8 z E 8 z 8 z E 8 ~D I CD C CD CJ ~D >' Z z c, z c, z z z z z ~ z O z c, z E z CD C ,~D ~D ~D ~D C CL~C C 'CL CL ~D
N.. ~ ~ ~, ~ CL ~ ~ CL ~ CL o~ ~D
O : : , ~
G CL CL O O CL CL CL o o E~, m O O O 0~ o m O m O m O m O o~
~ C
m ~ ~E Z c, ~ z z z T T 1 T <~ A 4 4 T
CD ~ 0 00 _ N
2 1 5 ~ ~ 1 2 - 13 8 - PCT/US94/02283 -- -- -- C~J N C'~
E N C') ~ N ~ N
T I T
C~ O O O O O
D
'` O ~ 'I~
E e 0 ~ ~-O O U~ (.) ~u2 ' 2 ' 2 ' ~ ~ a Z c~ O Z ~ z E ~ Z z O I . O x, x, ~' O
-- -- '~ ~ 'D . 2 _ N
8 o 8 8 8 8 m N
Z C Z Z Z Z O
E 5~ g~ '~ ~ ~ o y T
ooooooooooooo N . N . N ~ N . N N N Q N . N . N . N N . N Q
O O O O O O O O O O O O O
U~ C ~ C "~ S ~ C ~ C ~
~ Q r~ Q c~) Q c~ . ~ Q C~ Q ~ Q ~ Q ~ Q ~ Q ~ G ~ Q ~ Q
O O O O O O O O O O O O O
~ ' ~ ' ' ' c ' Q 'Q Q Q Q Q Q Q
m O O O O O O O O O O O O O
m ZZZZZZZZZZZZZ
", ." ~
I
,c I I I I I I C) c Q
o oN ~ o U~ ~o ~ ~ a~ --o -- --N Cl~
c~ o Cmpd. A B 1 B 2 Z D E F G X K M N ~ o 514 NH NH CO p-hydro y phenyl O 3,5-d hOAY CO 2,641hYdrAY CO H 1 3 phenyi phenyl 515 NH NH CO p-hydroxy phenyl NH 3,5-dihydroxy CO 2-- LOAY6- CO H 1 3 phenyi hydroxy phenyi 516 NH NH CO p-hydroxy phenyl NH 3,5- ' hoAy CO 2- LOAY 6- CO H 1 3 phenyi hydroxy phenyl 517 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2-carboxy~ CO H 1 3 phenyi hydroxy phenyi 518 NH NH CO p-hydroxy phenyl O 3,5 ' hoAy CO 2,6-d' lhoA~y CO H 1 3 phenyi phenyi 519 NH NH CO p-hydroAy phenyl O 3,5-dihydroxy CO 2,6~' lhoA) CO H 1 3 phenyi phenyl 520 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2~arLoAy CO H 1 3 1' phenyi cyclohe: ,e O
521 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3 phenyl phenyi 522 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 1-hJdl~.A~r 2- CO H 1 3 phenyi naphthyi 523 NCH2 NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2-carboxy-6- CO H 1 3 phenyi phenyi hydroxy phenyi 524 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2,6-dichloro phenyl CO H 1 3 phenyl 525 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2-carboxy CO H 1 3 phenyl c)_luh- ,e u 526 NH NH SO2 p-methyl phenyl O 3,5-dihydroxy CO 2-carboxy~ CO H 1 3 phenyi hydroxy phenyi j~, 527 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2 I 'hOA1 6- CO H 1 3 phenyl hydroxy phenyi y - I I I I I I I I I I I I I
l O O O O O O O O O O O O O
N N e~l N _ N ~1 N 1~ N N C~ ~ W N C~
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O O O O .` O O O O O O O O
` ~ ~ ~-~ 5 ~ ~ ~ ~ 5~ ~ ~ ~ ~
O O O O O O O O O O O Z O
~ ~ ~ ~ ~ ~
C G C ~ C ~. C ~ Q c ~ C 0. Cl. ~ c~. G G G C C ~ C
m O O O O O O O O O O O O O
m Z Z Z Z Z Z Z Z Z Z Z Z Z
8 ~ 8 o ~
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S _ _ _ _ _ _ _ _ _ _ _ y T
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O O O I I O O O O O O O
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m T T T
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_ .
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.
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CCI I I I I I I I I I T
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N . N . N . N . N ~ N . C~ N C N . N ~D N ~O N N
_ 8 8 8 8 8 8 8 8 8 8 8 8 ~ ~ ~ X ~ X ~ X ~ ' X, ~ X ~
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0 ~ ~ N ~
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~ ~ N N N N . N . N . N N . N . N . N . N ~- ~D
O O O O O O O O O O O O O
;~ C ~ C ~ C ;~ C ;~ C g C ~. C ;5~ C 5~ C ~ C ;g C ;~ C ;~. ~
Z O O O O O O O O O O O O
C~ CL C ~. 'G ~G ~ ~ ~ c ~ C G G C ~ Q G 1~ ~ G
m O O O O O O O O O O O O O
T T I I I T I I I I I T
O ~ o ~ = = o o = = O o 6 = =
E ~ Q 0 æ ~ 0N ~ ~ 0 tO
2157~ 146-Y
C) ~ I I I I I I I I I T
N . N . N N . ~ N . ~ N . C W ~ a ~ a G N ~ _ N N
2 ~ ~ ~ '`
Y ~ UJ ~ U~ Y
~ ~ ~ ~ G ~ C~ ~ ~ ~ ~ ~ G ~ c~ ~ G e~
O O O O O O O O O O O O O
c c~ , . .
~ c c~ c G ~ C~
-m O O O O O O O O O O O O O
m z I I I I I T
I C ~ I O ~ I ~ c ~ c c E cl~ u~ u~ ~D ~D o uo y S
O O O O O O O O O O O O
N . Q N _ O. N . N _ C~ N . N ~ N ~ N . N . ~ t'~ . I
O O O O O O O O O O O
C u~ ~, s ~. s ~ ~ ~
CL ~ O ~ G ~ ~ ~ ~ ~ C ~ O. ~ ~ ~ O. ~ G
O O O O Z O O O O O
~ 'S. ~ ~ u CL C ~ ~ G C~ ' ~ 1~ t~ G
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m O o O O O O O O O
.
m Z Z Z Z Z Z Z Z Z Z Z Z
s ~ I ~ I~ I ~ I ~ I ~ I ~ N S 5~ ~ ~ N~
Z t~ Z c~ Z c Z C Z o. Z G Z G Z G Z ~ Z ~ Z c Z
C~
E o _ N t~ O
~ ~ _ _ _ _ _ _ _ -- -- ~D No ~D
WO 94/2006t PCT/US94102283 21 j~3 i~ - 148 - --Z _ _ _ _ _ _ _ _ _ ~ _ y T I I T I T T T
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rD ~ r, WO94/2~K2 PCT~S94/0~83 21~ ~12 The following, nonlimiting Examples illustrate preferred methods for preparing the compounds for use in the method of the present invention and the data demonstrating PKC
inhibitory activity of the Compounds used in the present invention.
EXAMPL~8 3-Acetyl~minohexahydro-l-phenylmethylazepin-2,~-dione A solution of hexahydro-l-phenylazepin-2,3,4-trione-3-oxime (1.23 g, 5mmol) in 4:1 acetic acid/acetic anhydride (20 ml) was treated with Raney nickel (Aldrich, one-half tsp) in a Parr bottle and subjected to hydrogenation for eighteen hours at 40-45 psi and room temperature. The mixture was carefully evacuated of hydrogen and filtered through Celite~. The filter pad was then washed with methanol (with care taken not to let the filter pad become dry). The filtrate was concentrated in vacuo and the residue was diluted with toluene and further concentrated to remove most of the acetic acid. The residue was chilled on an ice bath and treated with saturated sodium bicarbonate carefully to avoid excessive bubbling. The cloudy aqueous solution was extracted with methylene chloride (3 x 50 ml), and the combined organic solution was dried (Na2SO4) and concentrated in vacuo. The residue was flash chromatographed on silica gel (eluted with 19:1 methylene chloride/methanol) to afford 3-acetylaminohexahydro-1-phenylmethylazepin-2,4-dione (1.11 g, 81~) as a white solid.
syn-3-Aminohexahy~ro-~-hy~roxy-l-phenylmethylazepin-2-one A solution of 3-acetylaminohexahydro-1-phenylmethylazepin-2,4-dione (0.82 g, 3.0 mmol) in absolute ethanol (lS ml) was treated with sodium borohydride (0.23 g, 6 mmol) and stirred for thirty minutes. The solution was then treated with water (5 ml) and concentrated in vacuo, and taken up in 2:1 ethanol/water (7.5 ml). Concentrated hydrochloric acid (2.5 ml) was added, and the mixture was refluxed for two hours and partially concentrated, then diluted with water (25 ml). The aqueous acidic mixture was extracted with ether (25 W094l20062 PCT~S94/02283 ml), and the aqueous solution was basified with 30% sodium hydroxide and extracted with methylene chloride (3 x 40 ml).
The combined methylene chloride extracts were washed with water (25 ml), dried (Na2S04), and concentrated in vacuo, to a yellow solid, which was recrystallized from ethyl acetate to afford syn-3-aminohPYAhydro-4-hydroxy-1-phenylmethylazepin-2-one (0.42 - g, 60%) as a white solid.
syn-3-Aminohexahydro-~,-hydroxy-l-phQnylmethylazepine A cooled (S-C) solution of lithium aluminum hydride/tetrahydrofuran (Aldrich, 1Ø N, 5.1 ml) under nitrogen was treated with syn-3-aminohexahydro-4-hydroxy-1-phenlmethylazepin-2-one (0.40 g, 1.7 mmol) in portions so that the pot temperature did not exceed 15-C. The mixture was refluxed for 6.5 hours, cooled on an ice bath, and carefully treated with water (0.21 ml), 15% sodium hydroxide (0.21 ml), and water (0.63 ml). The suspension was allowed to stir for five days, during which time the product partially decomposed (optimal time is 2-5 hours). The suspension was filtered, and the filtrate was concentrated in vacuo and chromatographed on silica gel (eluted with 90:8:2 methylene chloride/methanol/triethylamine). The appropriate fractions were concentrated in vacuo to afford syn-3-aminohexahydro-4-hydroxy-1-phenyl-methlazepine (0.22 g, 58%%) as a colorless oil.
syn-~exahydro-~-hydroxy-3-(~-phenylmethoxy)benzoyl~mino-1-phenylmethyl~zepine A solution of 4-benzyloxybenzoic acid (0.183 g, 0.8 mmol) in anhydrous tetrahydrofuran (2 ml) and N, N-dimethylformamide (0.5 ml) was treated with N,N'-carbonyldiimidazole ((0.15 g, 0.9 mmol) and stirred at roomtemperature for l.Sh. The solution was treated with syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.20 g, 0.9 mmol) in anhydrous tetrahydrofuran (1 ml). The mixture was stirred for eighteen hours, and then concentrated in vacuo.
the residue was taken up in lN sodium carbonate (20 ml), and W094l2~62 PCT~S94/02283 21~7~12 - 162 -the aqueous mixture was extracted with toluene (2 X 25 ml) containing a small amount of 2-propanol. The combined organic extracts were dried (Na2SO4) and the concentrated residue was flash chromatographed on silica gel (eluted with 3:1 ethyl acetate/hexane) to afford syn-hexahydro-4-hydroxy-3-(4-penylmethoxy)-benzoylamino-l-phenylmethylazepine (0.17 g, 50%) as a viscous oil.
3,5-Bis~phenylmethoxy)-~-~2-phenylmethoxybenzoyl benzoic ~ci~
ester with syn-Hexahy~ro-~-hydroxy-3-~4-phenylmethoxy benzoyl-~mino-l-phenylmethylazepine A solution of 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)benzoic acid (0.245 g, 0.45 mmol) in anhydrous methylene chloride (1.5 ml) was treated with N,N-dimethylformamide (two drops), then with 2 N oxalyl chloride/methylene chloride (Aldrich, 0.30 ml, 0.60 mmol), and stirred for one hour under nitrogen. The solution was concentrated in vacuo and placed under high vacuum for one hour. syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoyl-amino-l-phenylmethylazepine (0.170 g, 0.40 mmol) was dissolved in anhydrous methylene chloride (3 ml), treated with 4-dimethyl-aminopyridine (0.01 g) and triethylamine (0.12 ml, 1.2 mmol), and cooled in an ice bath under nitrogen. The acid chloride was removed from high vacuum and dissolved in anhydrous methylene chloride (2 ml), and was then added to the cooled solution. The mixture was allowed to warm to room temperature, stirred for one hour, and was partially concentrated in vacuo. The residual solution was deposited on a silica gel column and eluted (first with 2:1 hexane/methylene chloride, then with 1:1 hexane/methylene chloride) to afford (after concentration of the appropriate fractions) 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)- benzoic acid ester with syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepine (0.29 g 77~) as a white foam.
N O 94/20062 21~ 7 412 PCTrUS94tO2283 syn- (~- (3,5-Dihydroxy-~-(2-h~.osybenzoyl)benzoyloxy))hexhy~ro -3-(~-hydroxybenzoylamino)azepine A cloudy suspension of 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl-benzoic acid ester with syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepine (0.29 g, 0.30 mmol) in reagent ethanol (25 ml) was treated with - Pd(OH)2/C (Aldrich, Pearlman's catalyst, 0.20 g, 20%) in a Parr bottle, then subjected to hydrogenation for twenty four hours at 50-52 psi. The hydrogen was evacuated and the solution was carefully filtered through Celite~ under nitrogen, and the filter pad was washed with methanol (not to dryness). The filtrate was concentrated in ~acuo to crude material, which was flash chromatographed on a short column of silica gel (eluted with 1:1 CHClJEtOH) to afford a product (0.13 g) as a pale yellow foam. This was triturated from ether/ acetonitrile to afford syn-(4-(3,5-dihydroxy-4-(2-hydroxybenzoyl-benzoyl-oxy))hexahydro-3-(4-hydroxybenzoylamino) azepine (0.195 g, 62%) as a pale yellow powder (dihydrate): mp 177-179-C; Rf (1:1 CHCl3/EtOH on silica) 0.45; IR (KBr) 1623 cm , lH NMR (d6-DMSO) ~ 8.20 (d, lH, J = 8Hz), 7.10 (s, 2H), 7.02 (d, lH, J - 9 Hz), 6.91 (t, lH, J -= 8Hz), 6.78 (d, 2H, J = 9Hz), 5.39 (br d, lH, J = 7Hz), 4.48 (m, lH), 3.00-3.20 (m, 4H), 2.05-2.20 (M, lH), 1.70-2.00 (m, 3H). Anal. Calcd. for C27H26N2O8-2H20: C, 59.77; N,
carboxylic acids. In an alternative, preferred method, the alcohol is coupled with the acid chloride and treated with one equivalent of nBuLi which provides, upon rearrangement, the hydroxy ketone. Oxidation to the acid is effected in a two-step process using first pyridinium dichromate (PDC) or TEMPO
(2,2,6,6-tetramethyl-1-piperidinyloxy) followed by tetrabutylammonium permanganate (nBu4MnO4) or sulfamic acid (NH2SO3H) and sodium chlorite (NaClO2). The resultant acid is protected, for example, by conversion to the benzyl ester and then deprotected at the other ester position to provide the family of GFE carboxylic acids.
21~7 4~ 2 ~O 94/20062 PCT/US94/02283 Scheme IX B - 52-/=\ 1) BuLi ~ protect ~ 2) 5~ Iyl/J x ~CO2R
R'02C O R'02C O
~ dep~tec~, 5~CO2H
HO HO~
Br ~CO2R []
CIOC ~ CO2R ,KOtBu X
2) nBuLi HO2C O R'02C O
~ protect ~CO2R
R'02C O
depl~)tect ~CO2H
2~5 ~J~1~
W094/20062 rcTluss4lo2283 Scheme IXC provides a synthesis scheme for producing GFE carboxylic acids including the use of bromobenzenes as starting material. For example, a bromobenzene can be treated with n-butyl lithium and the acid chloride ester from scheme IXA to afford the benzophenone ester, which is deprotected to provide GFE carboxylic acids. The bromobenzene can also be treated with n-butyl lithium and the aldehyde ester from scheme IXA to give the diphenylcarbinol ester, which is either deprotected directly, oxidized with, for example, chromic acid and deprotected, or reduced with, for example, hydrogen and deprotected to provide the family of GFE carboxylic acids.
2157~1~?
WO 94/20062 PCT/US94/02283 _ Scheme ~X C
~r 1) ~'-r ~ CO R ~COzR
tCy~utc~L ~CO2H
L~.U...,L ~`CO2H
2) ~CO R ~CO2R
O ~ \
~CO2R ~CO2R
d~y~lt~ deprotect ~CO2H ~CO2H
S~I~UTE S! IEE~ (RULE 2~J
2157~12 WO9~/20062 PCT/US91102283 Scheme IXD provides a synthesis scheme for producing compounds according to the present invention including the use of halobenzenes, for example, bromobenzenes, and heterocyclic or cyclic compounds substituted with B1-B2-Z and DH, which are described in other schemes, as starting materials. For example, a bromobenzene can be reacted with a 4-heterobenzoate ester, in dimethyl formamide/triethylamine with heat or copper catalyst to give GFE acid chloride. This can be treated with heterocyclic compounds substituted with B1-B2-Z and DH in the presence of tetrahydrofuran/triethylamine to provide this family of balanoids.
PCT/US9~/02283 21~7~
Scheme ~X D
Arm 1 synthesis (continued):
DMF/Et3N
,Br HQ~ DMFlEt~N/Cu catalyst Q=O,~H,S
~3,a~ Ester ~ ~3`cocl HD~ B2--Z y ~CO
D~B, - B2--Z
Et3N/THF A
W094/20062 21~ ~ 4 ~ 2 rcTlus94lo2283 Scheme IXE provides a synthesis scheme f~or producing compounds according to the present invention including the use of aromatic or heteroaromatic halides protected, hetero-substituted aryl or heteroaryl acids or alcohols, and heterocyclic or cyclic compounds substituted with Bl-B2-X and DH, which are described in other schemes, as starting materials. For example, a bromoaromatic can be reacted with a protected 4-hetero-substituted aryl acid or alcohol in the presence of transition metal catalyst, for example, copper, and base to afford protected GFE acid or alcohol, which can be deprotected and treated with, for example, phosphorous penta chlorides to provide GFEX chloride. This is reacted with heterocyclic or cyclic compounds substituted with Bl-B2-Z and DH to provide another family of balanoids.
2 1 ~
- s8 -Scheme IX E
Cu catalyst G Br+ HF E XOR ~ G F E--XOR
or Pd(0) or Ni (0) E~3N
d~plotccL PCIS
G F E XOH
HD B~B2Z G F E--X\
~. D B1 Bz-Z
G--F--E XCI , \1~
E-,~IT~ / \
~A~)n W094/20062 2 1 5 7 ~ ~ ~ PCTIUS9~102283 Scheme IX F provides a preferred method for the preparation of the GFE-CO2H. Intermediate aldehyde is first protected, for example, as a cyclic acetal, which is then treated with nBuLi and DMF to afford aldehyde 1. Aryl bromide, is treated with n-butyl lithium and then aldehyde I to afford the alcohol. Oxidation using, for example MnO2 followed by acidic hydrolysis of the acetal gives the ketoaldehyde.
Oxidation to the acid using, for example, sodium chlorite and sulfamic acid, followed by the appropriate deprotection sequences affords GFE-CO2H.
wo 94/20062 PCT/US94/02283 2 1 5 ~ 60 -Scheme IX F
o o I Y I X~
/=1=\ 1) nBuLi ~/ ~ ~/~ 1) oxidation 2) 1 C~~ c/J~ 2) acid, H20 ~/~ 1) o~id~ion ~f/
2)protection /~1~
CO2R3)deprotection / CO2H
x2 x2 W094/20062 21~ 7 412 PCT~S9~/022~3 -Scheme IX G describes a preferred method for the preparation of GFE-CO2H where E is substituted with two OH or OR groups. Protected bis-phenol is treated with n-butyl lithium followed by aldehyde (prepared in Scheme IX F) provides, after oxidation with, for example, MnO2 the ketone intermediate. Following deprotection, the primary alcohol is oxidized, using, for example MnO2 followed by sodium chlorite/hydrogen peroxide. Hydrolysis of the acetal followed by treatment with MnO2, KCN, acetic acid and the alcohol R'-OH
provides the desired GFE-CO2H.
A detailed scheme for benzophenone synthesis is shown in Scheme IX H.
wo 94/2006~ - 62- PCT/US94/02283 Scheme IX G
O OCH3 y L~ l)nBuLi ~ OCH3 ~/r CHO O~ OCH3 o 3) oxidation o o OCH3 1) deprotection ~ H30+
2) oxidation ll o ! _ .
3) esterification ~ ~ / ~ CO2R
~ ~
o OCH3 O OCH3 Y ~ 1 1) KCN, AcOH /Y
~/~ ~MnO2, R'OH
CHO O/~2) dep~otection ~ ~ C02R' ~ CO2H
WO9~/2006~ 2 1 ~ 7 ~ l 2 - PCT/US9~/02283 Scheme IX H
Synthesis of MOM-Protected Benzophenone CO.M~ CO.Me ~OH
~3~ MOMCI ~ LAH, T~ ~
HO OH MOMO~OMOM MOMOJ~OMOM
OT~DMS
TBDMSCI r e ~
1~/10~10~0~10~1 HO nnO BnO
K,C03,BnBr ~ 1,3~ ,l).u~ H7~-C
CHO ~ PhMe,~ ~ o O~
HO hlOMO MOMO
f~q N.-lH, MOMCI ~ "BuLi, C6HI2 ~CHO
n ~MF ~
OTBDMS MOMlo hlOhlO~OTl~DMS
+ ~, I ''BuLi,0''C ~<
MOMO OhlOlM O J .1 ~
WO 9~/20062 PCT/US9 ~/022~3 2 1 ~
~ 64 ~
MOMO MOMO~,OMOM MOMO~OTBDMS
¢~0 OT~DMSh-- OMOM MnO~, CH,CI
o J "BuLi, 0"C. THF
O
MOMO~ OTBDMSMOMO~ OH
MOMO o ~ MOMO o ~ \\
~< TBAF, THF ~ MnO., CH.CI, OMOM > ~ OhlOM
O O
MOMO ~_~CHO hlOhlO~CO,hle MO~ KOH, 1, MO~
W~ OhlOlM MeOH. 0"C W~ OM()hl O O
NaCI02. HtO~ 18% H~SO~
N.aH2PO.~, MeCN-H.O a~l;orbe~ n SiO2 CH2( 12 MOh~O CO~H hlOlMO CO~Me MO~ MO~
~ OhlOhl W~CHO
WO 94/2006~
~1 J ~ ~ _ ~ PCT/US94/02283 ~,HO MOMO~,OMOM 'Blll i O"C B~ U~OTBDMS
~ ll l OMOM
OOTBDMS ~
o MlOhlO~OTBDMS MOMO ~--OH
MnO.. CH2CI- n,~ TBAF B~
W~ OMOM W~ OMIOMI
O O
MOMO~ ~CHO M~ lO~ CO2H
BnO O \ \\ BnO ~ \\
MnO-. CH-a2 ~< NsClO2, NaH2PO1 ~ <
H,02,MeCN-H.O l~'~~ OMOM
O O
2~ 2 - 66 -Scheme X provides means for produciAg compounds according to the present invention including the use of heterocyclic or cyclic compounds substituted with B1-Bz-Z and DXEFG where F is C=o, which are described in other schemes, as starting materials. For example, a heterocyclic or cyclic compound substituted with B1-B2-Z and DXEFG where F is C=O can be treated with a reducing agent such as sodium borohydride to afford balanoids where F is CHOH, or it can be treated with a sulfurizing agent, for example, phosphorus pentasulfide, to provide the family of balanoids with B2 and/or F equal to C=S.
~ro 9~/20062 21 S 7 412 PCTIUS9~102283 Scheme X
;~ G 1l HE~ G
I E
m=0-3 P4SIOor Lawesson's reagent 111 E~G
DC ll3 B~ C_ z A
X~,X2,X3 in~cpe~ ly = O or S
2 1 J7 '~12 - 68 -Scheme XIA provides a synthesis scheme for producing compounds according to the present invention including the use of heterocyclic compounds substituted with B1-B2-Z and DXEFG
where A is NH, described in other schemes, as starting materials. For example, a heterocyclic compound substituted with Bl-B2-Z and DXEFG where A is NH can be treated with an alkylating agent, sulfonylating agent, or acylating agent, for example acetyl chloride, in the presence of base to provide the family of balanoids with a substanted nitrogen.
In an alternative approach, Scheme XI B, the R group can be appended at an earlier stage in the synthesis using appropriately protected intermediates. For example, heterocyclic intermediate can be treated with an alkylating agent, sulfonylating agent or acylating agent, in the presence of base to provide the intermediate where Rl is not hydrogen.
2 PCT/US9 ~/02283 Scheme XI A - 69 -G--F--E--X-D~--B2--Z G--F--E--X-D 31-3z--Z
n( )m Et3N n(~ )m H Rl m=1-4,n=14 Rl=lower alkyl, aryl, or JR2 where m+n~or=5 J=CO,CN, or SO2 and R2=lower alkyl, aryl, alkylamino, or alkoxy Scheme XI B
HO~B1 -B2 ~Z ~B1 -B2 ~Z
N Et3N N
H Rl as described )--( in Scheme I Rl 2157~1~
WO9~12006' PCT/US94102283 Scheme XII provides a synthesis scheme for producing compounds according to the present invention including the use of cyclic or heterocyclic compounds substituted with B1-B2-Z and DXEFG where X is C=O, which are described in other schemes, as starting materials. For example, a heterocyclic compound substituted with B1-B2-Z and DXEFG where is C=O can be treated with an alkylating agent, for example dimethyl sulfate and an alcohol, such as methanol, to give the intermediate ketal.
This can be reacted with an organometallic, for example, the lithium salt of diethyl malonate, to provide the family of balanoids in which x is C=CR3R4. The ketal can be reacted with a primary amine, for example, butylamine, to provide the family of balanoids in which X is C=NR2.
WO 94/2006~ 7 - - 71 - ~ cL ~ PCT/US911022S3 X--\
E /
U~
a X E X E
O~
G U U
C~
oN
_ o C~.~
m c E ~X E/
O=~
~ U
~VO91/20062 PCT/US91/02283 57 41~ Scheme XIII provides syntheses for producing compounds according to the present invention including the use of GFE
carboxylic acids and cyclic or heterocyclic compounds substituted with B1-B2-Z and DH or B,H and DXEFG, which are described in other schemes, as starting materials. For example, a GFE carbinol can be treated with mesyl chloride and an amine base, such as triethylamine, followed by treatment with an iodide source such as sodium iodide to afford a GFE
methyl iodide. This can be reacted with a cyclic or heterocyclic compound substituted with Bl-B2-Z and DH in the presence of base such as sodium hydride to provide the family of balanoids in which X is CH2.
The cyclic or heterocyclic compound substituted with B,H and DXEFG can be reacted with a Z sulfonyl chloride, for example, benzenesulfonyl chloride, in the presence of base to provide the family of balanoids in which B2 is SO2.
N
o~ I
U
U ~ ' ~,u z ~ m E
s z I ' X~s X
u ~ U Z
E
U~ .
21~7~ 74 -Scheme XIV A provides a synthesis scheme for producing intermediates which are heterocyclic compounds substituted with Bl=NH2 and D=OH and in which A is oxygen. For example, a cyclic diene such as cyclopentadiene can be treated with a peracid such as peracetic acid and the epoxide opened with an amino species, for example, (dibenzylamino) dimethyl aluminum, followed by butyldimethylsilyl chloride to obtain the protected amino alcohol. This can be ozonolyzed and reduced to the diol, which is treated with tosyl chloride to give the chloro tosylate. Treatment with bis(trimethyltin)oxide or preferably potassium superoxide and a crown ether, for example, 18-C-6, provides the chloroalcohol which is ring closed on treatment with a strong base, such as methyllithium or butyllithium to afford the protected heterocyclic compound, which is deprotected to heterocyclic compounds substituted with B~=NH2 and D=OH and in which A is oxygen.
~'O 91/2006' 5 2 1 5 7 4 1 ~ PCT/US9 ~/02283 Scheme XIV A
1) Bn2NAIMe2 NBn2 Ac02H ~ 2) TBDMS-CI ~
1-3 1-3 ~ 'OTBDMS
1) 03 OTBDMS OTBDMS
2) NaBH4 Ts-CI .
HO~OH ~ Cl ~OTs NBn2 NBn2 2) BuLi, toluene, heat TBDMSOs NBn2 [H] TBDMSOs NH2 n(i~)m n(~ )m F- HOs NH2 n(~ )m 2 1~7 ~ ~/20062 - 76 - PCT/US9~/02283 Scheme XIV B provides a synthesis scheme for producing intermediates which are heterocyclic compounds substituted with B1=NH2 and D=OH and in which A is sulfur. For example, a cyclic diene such as cyclopentadiene can be treated with a peracid such as peracetic acid and the epoxide opened with an azide species, for example, sodium azide, followed by t-butyldimethylsilyl chloride to obtain the protected amino alcohol. This can be ozonolyzed and reduced to the diol, which is treated with mesyl chloride to give the dimesylate and ring closed with, for example, bis(trimethyltin) sulfide or preferably, lithium sulfide and an amine base such as triethylamine. Reduction of the azide and deprotection occurs on treatment with lithium aluminum hydride to give heterocyclic compounds substituted with Bl = NH2 and D = OH and in which A
is sulfur.
Compounds where A is SO2 can be prepared from compounds where A is sulfur by treatment with an oxidizing agent, for example, peracetic acid,followed by deprotection to provide compounds wherein A is SO2 (Scheme XIV C).
WO 94/20062 21 S 1 ~ ~ ~ PCT/US91/02283 Scheme XIV B
1) NaN3, NH4CI, CH30H/N3 Ac02H ~< 2) TBDMS-CI
1-3 1-3 ~ "OTBDMS
1) 03 OTBDMS OTBDMS
2) NaBH4 . Ms-CI
HO~OH ~ MsO ~OMs N3 NBn2 1 ) Li2S, Et3N, CH30H HO NH2 2)LiAlH4 S
-'O 9~/~006' Scheme X~V C - 78 - `-215~
/ /
___ 1) RCO3H ~ A
S~ )m 2) De~rut~c~ion n( ~ ~ )m O O
~ro94/20062 215 ~ ~1~ PCT/US94/02283 Scheme XV provides a synthesis scheme for producing compounds according to the present invention including the use of Z sulfonyl chlorides, such as benzenesulfonyl chloride, and cyclic or heterocyclic compounds substituted with Bl=NH2 and DH, which are described in other schemes, as starting materials.
For example, a Z sulfonyl chloride and a cyclic or heterocyclic compound substituted with Bl=NH2 and DH are combined in the presence of base to afford the sulfonamide, which is reacted with GFEX halide to provide the family of balanoids in which B
is N and B2 is SO2.
WO 94/2006' PCT/US93/022S3 2 ~r'J - 80 -Scheme XV
ol o D NH2 Cl--S--Z D N--S--Z
m( ~ ) n Et~N m( ~ ) n G F E X~ H ¦¦
G - F E--X--Cl D N--S--Z
m( ~A~ ) n G--F E X~ R
D N_S--Z
K2CO3, R2X \~ ( o m( g~A~ ) n W094/20062 2 1 5 ~ ~ ~ ~? PCT/US9S/02283 Scheme XVI provides a synthesis scheme for producing compounds according to the present invention including the use of ketones, for example, acetophenone, and azepinediones, such as 1-benzylazepin-2,4-dione, as starting materials. For example, the ketone can be brominated with bromine in acetic acid and protected with ethylene glycol to afford the bromomethyl ketal, which is added to a base-treated solution of the azepinedione to give the alkylated azepinedione. This is reduced to the hydroxyazepine with, for example, lithium aluminum hydride, and reacted with base and GFEX halide, which is described in other schemes, then deprotected to provide the family of balanoids where Bl is CH2, Bz is C=O, and D is O.
These compounds can be reacted with, for example, phosphorus tetrasulfide, to provide the family of balanoids where Bl is CH2, B is C=S and D is O.
WO 94/2006' 2 1 5 ~ 82 -Scheme XVI
--'6Z Br2, HOAcBr~_~Z HOCH2CH20H, Br 7~
O o pTsOH, Benzene, O J
Dean-Stark Trap ~,~ 1 ) NaH ~ LIAIH~
G--F E X
OH O
(~<Z 1.) NaH (~<Z
N \J 2-~ Cl X E--F--G N \_l Rl R
G--F E X
1.) PPTS, Acetone H20 , (~' P4S, o N
G--F E X\
o (~Z
N S
-'09~/20062 ~ 2 PCT~S9~10228 Scheme XVII provides a synthesis scheme for producing compounds according to the present invention including GFE
carboxylic acids, which are described in other schemes, and cyclic or heterocyclic groups substituted with OH and BlB2Z, also described in other schemes, as starting materials. For example, a GFE carboxylic acid can be treated with oxalyl chloride and N,O-dimethylhydroxylamine to afford the methoxymethyl amide, which is reacted with a methyl organometallic to give the methyl ketone. This is then deprotonated with base and reacted with the product from treatment of a cyclic or heterocyclic group substituted with DH
and BlB2z with tosyl chloride to provide the family of balanoids with D as CH2 and X as C=O.
WO 9~/2006~
- PCT/US9~/022~3 2157~12 Scheme XVII
?
) (COC1~2/D~
CH2Cl2 ~'.~"
G F--E--CO2H ~ G F--E--CON(OMe)(Me) 2) MeNHOMe M=Li, MgBr G F E--COCHa HO B~ B2Z TsCI TsO B~ B2Z
(~)n ~ (~) LiN(SiMe3)2 G--F E--COC~Bl B2Z
(b~A~)n W094/2006~ 215 i ~ CT/US94/02t~3 Scheme XVIII provides a synthetic scheme for compounds of the invention where K is not equal to H. Oxidation of the alcohol using, for example, oxalyl chloride and dimethylsulfoxide (Swern oxidation) followed by addition of an organometallic reagent, for example, the complex of trimethylaluminum and methyl magnesium chloride to afford the tertiary alcohol. This intermediate is converted using the Scheme I to compounds of the invention.
wO 9~/2006~
Scheme XVIII
2157~12 HO B1 - B2--z 1 ) oxalyl chloride HO ~ B1 - B2--Z
)~ DMSO, Et3N )~
A/~ 2) Me3AI, MeMgCl Q\A/Q
as described Targets where K = CH3 in Scheme I
~094/2006~ 21 S 7 41~ PCT/US94/02283 Scheme XIX provides a synthetic scheme for compounds of the invention where the group G is substituted with an alkoxycarbonyl group. Compounds wherein G is substituted with a carboxy group are treated with an alkylating agent, for example, methyl iodide, and a base such as sodium carbonate to provide the target compounds.
WO 9~/20062 - 88 - PCT/US9~1/022~3 ~cheme ~lX
C~X~
X D K B IB2Z R-halo ~A~ )m K2CO3, DMF
OH
Y I X
X~
x2 DK B IB2Z
n(~ )m where R = alkyl, substit-1tPd alkyl wo 9~,2006? 2 1 S 7 4 1 2 PCT/US9~/02283 Scheme XX provides a synthetic route for compounds of the invention where G and/or E residues are substituted with acyloxy groups. Target compounds, which possess one or more hydroxyl groups on G or E is treated with an acylating agent, for example, acetyl chloride, ethyl chloroformate, and the like, in the presence of a base such as pyridine or triethylamine to provide the target compounds.
WO 94/2006 2 PCTIUS9 ~/022~3 Scheme XX ~ 90 ~
lS~ 41~ o OH HO~ X 1 ) _1l D K B IB2Z R Cl pyridine ~A~ 2) dep~~ L
~O~o~X~ ~
o~L O~ D~ B~B2Z
n(~A~ )m ~'O9~/20062 21~ 7 ~12 PCT/US9~/022X3 Scheme XXI provides a synthetic scheme for the preparations of compounds where Bl is N-R and B2 is C=O.
Intermediate amide, following protection of the D
functionality, is treated with a strong base such as KOtBu or KH and an alkylating agent such as methyliodide or dimethylsulfate to provide the intermediate where B1 is N-R.
This can be converted using the procedures outlined in Scheme I to afford the target compound.
WO 9~/2006~
2 PCT/US9`1/02283 Scheme XXI ~ 9 HD K N~ 1) Protection HD K N~
\)~( o 2) strongbase, \~4 0 ~ A/~ alkylating agent as described target where R = lower alkyl in Scheme I
Scheme XXII describes the synthesis of compounds of the invention in which group G is substituted with a tetrazole ring. Keto aldehyde (prepared as described in Scheme IX F) is treated with hydroxylamine hydrochloride in dimethylformamide to provide the nitrile. Following deprotection to the acid, it is coupled to provide target compounds wherein G is substituted with a nitrile group. Treatment with trimethylsilylazide and nBu2SnO followed by deprotection affords the target compounds.
WO 91/20062 PCT/US9~/02283 Scheme ~Xll 94 ~
1 HONH2-HCl ~9/l----¢~/
~CHO 0~CO2R CN /J~CO2R
t~D K B ,B2Z CN 5~ ~
~4 X~ D K B ,B2Z
n(~ )~ )m n(~)m ) TMSN3 Y ll x' nBu2SnO ~/~
2) Deprotect ~
D K B~B2Z
n(~A~ )m WO9~/2006' 21~ 7 ~12 PCT/US9~/02283 Scheme XXIII provides a synthetic scheme for the preparation of compounds where D is N-R. Amine intermediate (prepared as described in Scheme III) is converted to the trifluoroacetamide. Treatment with a strong base, such as KOtBu and an alkylating agent such as methyl iodide or dimethylsulfate followed by cleavage of the trifluorocetamide provides the intermediate amine wherein D is N-R. This is converted to compounds of the invention using procedures outlined in Scheme III.
WO 9~/20062 n~ PCT/US9~1/02283 Scheme XXIII - ~u- -Z157412 o 1~
\ HN Bl-B2--Z 1) KOtBu n(~ )m 1 ) TFA, CDI )_~ (CH3)2S04 g\ A/~ 2) KOH, CH30H
~ CH3 as described , Targets where D = -NCH3 n(~ ~A/Q)m in Scheme I
Persons of ordinary skill in the art will recognize that the foregoing schemes are exemplary only and should not be construed to be limiting.
Pharmaceutical preparations incorporating compounds according to the present invention can be used to block PKC
activity related to abnormal or undesirable cellular events and activity including tumorogeneis and cellular activity related to inflammation and reperfusion injury. Treatment of disorders and disease conditions can be performed by administration of effective amounts of pharmaceutical preparation that comprise compounds according to the present invention. Compounds can be formulated for human and animal prophylactic and therapeutic applications by those having ordinary skill in the art. The range of amounts of a compound to be administered to mammals, lS particularly humans, to be effective in inflammatory, tumor or reperfusion injury therapy can routinely be determined by those having ordinary skill in the art.
The compounds and pharmaceutical compositions of the invention may be administered by any method that produces contact of the active ingredient with the agent's site of action in the body of a mammal or in a body fluid or tissue.
These methods include but not limited to oral, topical, hypodermal, intravenous, intramuscular and intraparenteral methods of administration. The compounds may be administered singly or in combination with other compounds of the invention, other pharmaceutical compounds such as chemotherapeutic compounds, or in conjunction with therapies such as radiation treatment. The compounds of the invention are preferably administered with a pharmaceutically acceptable carrier selected on the basis of the selected route of administration and standard pharmaceutical practice.
The compounds of the invention are administered to mammals, preferably humans, in therapeutically effective amounts which are effective to inhibit protein kinase C, to inhibit tumor cell growth, inhibit inflammation of tissue, inhibit keratinocyte cell proliferation, inhibit oxidative burst from neutrophils or inhibit platelet aggregation. The W094/2006' PCT/US94/02283 2157~1~ I 98 -dosage administered in any particular instance will depend upon factors such as the pharmacodynamic characteristics of the compound of the invention, its mode and route of adminis-tration; age, health, and weight of the recipient; nature and extent of symptoms; kind of concurrent treatment, frequency of treatment, and the effect desired.
Compounds according to the present invention inhibit the activity of PKC in cells. The range of the amount of inhibitory compound that is effective for inhibiting PKC
activity can be determined by one having ordinary skill in the art. By inhibiting PKC activity, balanoids are useful in the treatment of disease conditions in which control of cellular growth, regulation and/or differentiation is desirable. An effective amount of a balanoid can be administered to mammals who are suffering from inflammatory, cardiovascular or neoplastic diseases, particularly inflammation, reperfusion injury and cancer, in order to counter the disease at the cellular level.
It is contemplated that the daily dosage of a compound of the invention will be in the range of from about 1 ~g to about lOO mg per kg of body weight, preferably from about 1 ~g to about 40 mg per kg body weight, more preferably from about lO ~g to about 20 mg per kg per day.
Pharmaceutical compositions of the invention may be administered in a single dosage, divided dosages or in sustained release forms. Persons of ordinary skill will be able to determine dosage forms and amounts with only routine experimentation based upon the considerations of this inven-tion. Isomers of the compounds and pharmaceutical compositions, particularly optically active stereoisomers, are also within the scope of the present invention.
The compounds of the invention may be administered as a pharmaceutical composition orally in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions. The compounds may also be administered parenterally in sterile liquid dosage forms or topically in a carrier. The compounds of the ~094/20062 2 1 5 7 ~ 1 2 PCT/US94/02283 _ 99 _ invention may be formulated into dosage forms according to standard practices in the field of pharmaceutical preparations.
See ~emington's Pharmaceutical Sciences, A. Osol, Mack Publishing Company, Easton, Pennsylvania.
Compounds of the invention may be mixed with powdered carriers, such as lactose, sucrose, mannitol, starch, cellulose derivatives, magnesium stearate, and stearic acid for insertion into gelatin capsules, or for forming into tablets. Both tablets and capsules may be manufactured as sustained release products for continuous release of medication over a period of hours. Compressed tablets can be sugar or film coated to mask any unpleasant taste and protect the tablet from the atmosphere or enteric coated for selective disintegration in the gastrointestinal tract.
Liquid dosage forms for oral administration may contain coloring and flavoring to increase patient acceptance, in addition to a pharmaceutically acceptable diluent such as water, buffer or saline solution.
For parenteral administration, a compound of the invention may be mixed with a suitable carrier or diluent such as water, an oil, saline solution, aqueous dextrose (glucose) and related sugar solutions, glycols such as propylene glycol or polyethylene glycols. Solutions for parenteral administration contain preferably a water soluble salt of the compound of the invention. Stabilizing agents, antioxidizing agents and preservatives may also be added. Suitable antioxidizing agents include sodium bisulfite, sodium sulfite, and ascorbic acid, citric acid and its salts, and sodium EDTA.
Suitable preservatives include benzalkonium chloride, methyl-or propyl-paraben, and chlorbutanol.
Animal studies have shown that perhaps 50% or more of ischemic-related myocardial damage can be attributed to polymorphonuclear leukocytes (neutrophils) which accumulate at the site of occlusion. Damage from the accumulated neutrophils may be due to the release of proteolytic enzymes from the activated neutrophils or the release of reactive oxygen intermediates (ROI). Much of the "no reflow" phenomenon 2,15~12 - 100 -associated with myocardial ischemia is attributed to myocardial capillary plugging. The plugging of capillaries has been attributed to both aggregated platelets and aggregated neutrophils. Although both cell types are aggregate during the ischemic event, the relative contribution of each to capillary plugging has not yet been established. It is accepted that the damage by neutrophils to myocardial tissue proceeds through a cascade of events, one of the earliest being the bonding of activated neutrophils to damaged vascular endothelium. However the binding of the neutrophils is significantly enhanced by their activation. Thus, an even earlier event is the generation of molecules (such as cytokines, and chemotactic factors) which can function as activation stimuli. These molecules probably originate from damaged and aggregated platelets, from damaged vascular endothelium, or from the oxidation of plasma proteins or lipids by endothelial-derived oxidants.
Strategies for overcoming the deleterious effects of reactive oxygen intermediates have centered on the development of scavengers for the molecules. Superoxide dismutase (SOD) has been shown to be a particularly effective scavenger of superoxide, but suffers from a very short half-life in the blood. Several companies have approached this problem by creating versions of the enzyme with increased half-lives by techniques such as liposome encapsulation or polyethylene glycol conjugation. Reports on the effectiveness of these new versions are mixed. Catalase, a scavenger of hydrogen peroxide, and hydroxyl radical scavengers have also been tested and found to be effective to varying degrees. However, none of the strategies designed to scavenge reactive oxygen intermediates will prevent the aggregation of platelets, the release of chemotactic molecules, the activation and adherence of neutrophils to vascular endothelium, or the release of proteolytic enzymes from activated neutrophils.
One advantage of protein kinase C inhibitors as therapeutics for reperfusion injury is that they have been demonstrated to: 1) block platelet aggregation and release of W094l20062 21~ 7 ~12 PCT/US94102283 neutrophil activating agents such as PAF; 2) block neutrophil activation, chemotactic migration, and adherence to activated or damaged endothelium; and 3) block neutrophil release of proteolytic enzymes and reactive oxygen intermediates. Thus, these agents have the capability of blocking all three of the most significant mechanisms of pathogenesis associated with reperfusion injury and should thus have a decided therapeutic advantage.
The table below contains a list of compounds according to the present invention. For convenience, in some cases the substituient groups described in their noun form rather than as adjectives, e.g. pyridine rather than pyridyl. It is to be understood that, unless specified, points of attachment of these functional groups can be any which are typically found in organic chemistry. Thus, for example, recitation of "pyridine"
as a substituient comprehends attachment at the 2, 3, or 4 position .
Cmpd. A 81 ~ 2 Z D E F G X K m n NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3 phenyl 2 NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3 phenyl 3 NH NH SO2 p-methyl phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3 phenyl 4 NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO phenyl CO H 1 3 phenyl CH2 NH CO p-hydroxy phenyl 0 2,6-dihydroxy CH2 phenyl CO H 1 3 phenyl 6 CH2 NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3 phenyl 7 CH2 NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-carboxy phenyl CO H 1 `3 phenyl 8 NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-carboxy phenyl CO H 1 3 phenyl 9 NH NH CO pyridine 0 3,5-dihydroxy CO 2-- I,UAy 6- CO H 1 3 phenyl hydroxy benzene NH NH CO pyrrole 0 3,5-dihydroxy CO 2-~.~ b~)A~6- CO H 1 3 phenyl hydroxy phenyl 11 NH NH CO oxazole 0 3,5~" 'hoA~ CO 2-- bUAY 6- CO H 1 3 phenyl methyl phenyl 12 NH NH CO indole 0 3-hydroxy-5- CO 2-hydroxy-6- CO H 1 3 r `h~l~ h_.,~1 methylphenyl 13 NH NH CO purine 0 3 1 `h y 5- CO 2,6-dimethyl CO H 1 3 i~
hJII h~ l phenyl Cmpd. A 8 1 ~ 2 ¦ D E F ¦ G ¦ X K ¦ m n 14 NH NH CO ~uran O 3-hydroxy CO 2,6-~tn hvAy CO H 1 3 phenyi phenyi NH NH CO lI,k~i-,_.,e O 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3 phenyi phenyi 16 NH NH CO PJ-i ' ' ,e O 3,5~1n 'hGA~ CO 2~ L.oAf 6- CO H 1 3 phenyl hydroxy benzene 17 NH NH CO ,~".i ,hli.,e O 3-hydroxy-5- CO 2~arboxy4- CO H 1 3 . . ._lhJ4 1 ~i hydroxy benzene 18 NH NH CO pyrazine O 3.5~' '~ Y CO 2~ IJVA~ 6- CO H 1 3 phenyi hydroxy benzene 19 NH NH CO imidazole O 3-hydroxy-5- CO 2~ LVA~r 6- CO H 1 3 ~
... `hy4h_.,~1 hydroybenzene ~ i--NH NH CO Ihiazole O 3,5-dihyroxy CO 2- I,oAf 6- CO H 1 3 ~ CJ~
benzene hydroxy benzene w 21 NH NH CO isoxazole O 3,5-d' . IhuAy CO 2~ boAy 6- CO H 1 3 ~ i~
phenyi hydroxy benzene l~
22 NH NH CO pyrazole O 3,5~dihydroxy CO 2- bvAy 6- CO H 1 3 phenyi hydroxy benzene 23 NH NH CO i~ r~l',' 'e O 3,5-dihydroxy CO 2~- LUA~6- CO H 1 3 benzene hydroxy benzene 24 NH NH CO benzene O 3,5~" . hvAy CO 2~ bOAY 6- CO H 1 3 phenyi hydroxy phenyl NH NH CO methyl benzene O 3-hydroxy-5- CO 2,6-dimethyl CO H 1 3 ~?
,.. _lh,1~ h_.. ~1 phenyi r~
26 NH NH CO dimethyl benzene O 3,5~ ./ CO 2,6~di,.. _1hvAy CO H 1 3 0 phenyl phenyi i~-27 NH NH CO trimethyi benzene O 3,5-dihydroxy CO 2~arboxy-6- CO H 1 3 benzene hydroxy benzene ~.:~ o Cmpd. A dl ¦ ~ Z ¦ D E F ~I ¦ X K m ¦ n ~ æ
28 NH NH CO Mr~.. h~1 0 3-hydroxy-5- CO 2~arboxy~ CO H 1 3 hyl~,h~" /l hydroxy benzene 29 NH NH CO ethyl benzene 0 3,5-dihydroxy CO 2-carboxy~ CO H 1 3 benzene hydroxy phenyl NH NH CO tetraethyl benzene 0 3-hydroxy CO 2~ OAY 6- CO H 1 3 phenyl hydroxy benzene 31 NH NH CO propyl benzene 0 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3 phenyl phenyl 32 NH NH CO tetra propyl 0 3,5-dihydroxy CO 2-carboxy~ CO H 1 3 benzene benzene methyl phenyl 33 NH NH CO butyl benzene 0 3-hydroxy-5- CO 2-carboxy-6- CO H 1 3 methoxy phenyl hydroxy benzene 34 NH NH CO tetrabutyl benzene 0 3,5-dihydroxy CO 2~ I,oAt 6- CO H 1 3 benzene hydroxy benzene NH NH CO pentyl benzene 0 3,5-dihydroxy CO 2-carboxy-6- CO H 1 3 benzene hydroxy benzene 36 NH NH CO t~ benzene 0 3,5-dihydroxy CO 2,6t'. hùAy CO H 1 3 phenyl phenyl 37 NH NH CO methoxy benzene 0 3,5-dihydroxy CO 2,6-dimethyl CO H 1 3 benzene phenyl 38 NH NH CO di~ hUAy benzene 0 3,5~i,.. 4hUAt CO 2~ LOAY 6- CO H 1 3 phenyl methylphenyl 39 NH NH CO td~ thoAy benzene 0 3,5-dlhydroxy CO 2~ I~OAY 6- CO H 1 3 benzene methoxy phenyl u NH NH CO tetra methyl 0 3-hydroxy-5- CO 2-carboxy~- CO H 1 3 ~, benzene methoxy phenyl hydroxy phenyl x~
41 NH NH CO ethoxy benzene 0 3,5-dihydroxy CO 2; t~oAy 6- CO H 1 3 benzene hydroxy phenyl Cmpd ¦ A E1¦ d l Z ¦ D E F G X K m n 42 NH NH CO dlethoAy benzene 0 3-hydroAy-S- CO 2-- boA~ 6- CO H 1 3 methyl phenyi methoxy phenyi 43 NH NH CO nitro benzene 0 3,5-dihydroAy CO 2,6-dimethyl CO H 1 3 benzene phenyl 44 NH NH CO dinitro benzene 0 31 ~h y 5- CO 2~ bUAY 6- CO H 1 3 methyi phenyi hydroxy benzene NH NH CO halo benzene 0 3,5-dihydroAy CO 2~arboAy4- CO H 1 3 benzene hydroxy phenyi 46 NH NH CO dihalo benzene 0 3-hydroxy CO 2-carboxy4- CO H 1 3 phenyl hydroxy phenyi 47 NH NH CO trihalo benzene 0 3,5-dihydroxy CO 2-carboxy4- CO H 1 3 phenyl hydroxy phenyi I 1 4~ NH NH CO tetrahalo benzene 0 3,5-dihydroxy CO 2~ bUA~ 6- CO H 1 3 benzene methoxy phenyl --49 NH NH CO benzene_ bUA~1 0 3,5-~L. huAy CO 2-carboxy4- CO H 1 3 1 acid phenyi methoxy phenyi NH NH CO benzene 0 3,5-dihydroxy CO 2~arhoAy4- CO H 1 3 d;~IIIUAYI;~ acid benzene methyi phenyi 51 NH NH CO b~" .id~ amido 0 3 m 'h /5- CO 2-hydroxy4- CO H 1 3 benzene methyi phenyl methyi phenyi 52 NH NH CO benzene dbmide 0 3-hydroxy CO 2,6-dimethyi CO H 1 3 phenyi phenyi 53 NH NH CO phenol 0 3,5-dihydroxy CO 2,6-d;,,,~lhuAy CO H 1 3 c benzene phenyi u 54 NH NH CO dihydroxy benzene 0 3,5-dihydroAy CO 2,6~ihydroxy CO H 1 3 phenyi phenyl NH NH CO trihydroxy benzene 0 3,5-dihydroxy CO 2-carboxy4- CO H 1 3 benzene methoxy phenyi Cmpd. A dl E2 ¦ Z D ¦ E F G X K m n l~
56 NH NH CO pe... 'hJd~vA~ O 3hJd~uA~t5- CO 2~ LUA~6- CO H 1 3 benzene methyi phenyi methoxy phenyi 57 NH NH CO triethoAy benzene 0 3,5-dlhydroxy CO 2,6-dimethyi CO H 1 3 benzene phenyi 58 NH NH CO tetra ethoxy O 3-hydroxy CO 2,6-dihydroxy CO H 1 3 benzene phenyi phenyi S9 NH NH CO propoxy benzene 0 3,5-dihydroxy CO 2 carboxy~ CO H 1 3 benzene hydroxy phenyi NH NH CO ~ lU~UAy benzene 0 3n.- .y5- CO 2~arboAy~ CO H 1 3 methyi phenyi hydroxy phenyi 61 NH NH CO t~i~,u~oAybenzene O 3,5-dihydroxy CO 2-t~rboxy-6- CO H 1 3 benzene methyi phenyi 62 NH NH CO tetra propoAy O 3,5-dihydroxy CO 2,6-" hùA~t CO H 1 3 benzene phenyi phenyi tJ
63 NH NH CO amidine 0 3,5-dihydroAy CO 2 carboxy~ CO H 1 3 phenyi hydroxy phenyi 64 NH NH CO dbmino benzene 0 3,5-dihydroxy CO 2-1,/.' UAY 6- CO H 1 3 benzene methyi phenyi NH NH CO methoxy pyridine 0 3,5-dihydroxy CO 2-o bVA'Y 6- CO H 1 3 benzene methyi phenyi 66 NH NH CO di,.. _lhuAy pyndine 0 3-hydroxy-5- CO 2-n jJUXY 6- CO H 1 3 methoxy phenyi methoxy phenyi 67 NH NH CO hydroxy pyridine 0 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3 c benzene phenyi v 68 NH NH CO dihydroxypyridine O 3,5-dihydroxy CO 2,6t-i.. ,~ll.GAy CO H 1 3 benzene phenyi 69 NH NH CO ethoAy pyrrole 0 3,5-dihydroxy CO 2-hydroxy-6- CO H 1 3 phenyi methyi phenyi ¦Cmpd. A El ¦ E2 D E F G X ¦ K m n NH NH CO dihydroxy pyrrole 0 3,5-dihydroAy CO 2-carboAy4- CO H 1 3 benzene hydroAy phenyl 71 NH NH CO dim h~JAy Indole 0 3-hydroAy-S- CO 2~arboAy4- CO H 1 3 methoxy phenyl hydroxy phenyl 72 NH NH CO hydroAy purine 0 3,5-dihydroAy CO 2-. bUA~ 6- CO H 1 3 benzene me~hyl phenyl 73 NH NH CO ~ hUAy ~uran 0 3: ~h J5- CO 2~ bUAy 6- CO H 1 3 methyl phenyl me~hoAy phenyl 74 NH NH CO hydro)yll,k~h~.,e 0 3,5-dihydroAy CO 2,6-dime~hyl CO H 1 3 benzene phenyl NH NH CO melhoAy p~ e 0 3,5-dihydroAy CO 2.6-dihydroxy CO H 1 3 benzene phenyl 76 NH NH CO 'i~.. lhùAy O 3-hydroAy CO 2,6-~- hùAy CO H 1 3 ~ r~
~".iJ~;"e phenyl phe~nyl ~1 77 NH NH CO hydroxy~,.;'".e O 3,5-dihydroAy CO 2-carboAy4- CO H 1 3 1 -~
phenyl hydroAy phenyl 78 NH NH CO diamido pf~ . e O 3,5-dihydroAy CO 2-carboxy4- CO H 1 3 J~
benzene hydroxy phenyl 79 NH NH CO amido pyrazine 0 3,5-'.lhuA1 CO 2~ bùAt 6- CO H 1 3 phenyl methoAy phenyl NH NH CO dielho)ty pyrazine 0 3 5-dihydroAy CO 2-carboAy4- CO H 1 3 benzene hydroxy phenyl 81 NH NH CO p-hydroAy phenyl O pyndine CO 2-~rboAy4- CO H 1 3 c melhyl phenyl v 82 NH NH CO p-methoAy phenyl O pyrrole CO 2-hydroAy4- CO H 1 3 me~hyl phenyl 83 NH NH CO p-chloro phenyl O oxazole CO 2,6-~" huAy CO H 1 3 phenyl Cmpd. A E~ Ez Z D E F G X K m n ~, 84 NH NH CO~ hUA~ phenyi O indole CO 2,6-dihydroxy CO H 1 3 phenyi NH NH CO .. hJd~UA~ phenyi O purine CO 2- buAf 6- CO H 1 3 hydroxy phenyi 86 NH NH COm-chloro phenyi O furan CO 2-. buA~6- CO H 1 3 hydroxy phenyi 87 NH NH CO o-methoxy phenyi O Ihlu~ .. e CO 2-hydroxy4- CO H 1 3 methyi phenyi 88 NH NH CO o-hydroxy phenyi- O ~,.; ' ,e CO 2,6-dimethyi CO H 1 3 phenyi 89 NH NH CO o-chloro phenyi O ~".i ' .e CO 2,6-. . hùAy CO H 1 3 phenyi NH NH CO p-hydroxy phenyi O pyrazine CO 2-carboxy4- CO H 1 3 o methoxy phenyi I
91 NH NH CO m-methoAy phenyi O imidazole CO phenyi CO H 1 3 92 NH NH CO m-hydroxy phenyi O Ihiazole CO 2-carboxy4- CO H 1 3 methyi phenyi 93 NH NH CO p-methoAy phenyi O Isoxazole CO 2-carboxy4- CO H 1 3 hydroxy phenyi 94 NH NH CO o-methoAy phenyi O pyrazole CO 2-carboxy4- CO H 1 3 hydroxy phenyi NH NH CO m-hydroA~ phenyi O Is~tl,' 'e CO 2-carboxy4- CO H 1 3 hydroxy phenyi u 96 NH NH CO o-methoAy phenyi O benzene CO 2-carboxy4- CO H 1 3 o methyi benzene 97 NH NH CO o-chloro phenyi O methyibenzene CO 2-carboxy4- CO H 1 3 methyi benzene CmDd. A E1 E2 Z D E F ~ X ¦ K m n 98 NH NH C0 p-hydroxy phenyl O dimethyl CO 2~ bUA'y6- CO H 1 3 benzene hydroxy phenyl 99 NH NH C0 p-chloro phenyt O trlmethyl CO 2-carboxy-6- CO H 1 3 benzene me~hyl phenyl 100 NH NH C0 m-hydroxy phenyl 0 t- ... lh~ CO 2-l,JJ~uA~6- CO H 1 3 methyl phenyl 101 NH NH C0 ... m lhuAy phenyl O ethylbenzene CO 2,6-Ji~ hUAy CO H 1 3 phenyl 102 NH NH C0 p-hydroAy phenyl O tetraethyl CO 2-carboxy-6- CO H 1 3 benzene methoxy phenyl 103 NH NH C0 p-methoAy phenyl O propylbenzene CO 2.6-dihydroxy CO H 1 3 benzene phenyl 104 NH NH C0 m-hydroxy phenyl O tetrapropyl CO 2-hJJ~UAy6- CO H 1 3 0 ~-~
benzene methyl phenyl ~ ~n 105 NH NH C0 o-hydroxy phenyl O butyl benzene CO 2,6-dihydroxy CO H 1 3 phenyl 106 NH NH C0 m-hydroxy phenyl O tetrabutyl CO 2-carboxy-6- CO H 1 3 l~a benzene me~hyl benzene 107 NH NH C0 m-hydroxy phenyl O pentytbenzene CO 2-carboAy-6- CO H 1 3 methyl benzene 108 NH NH C0 o-hydroxy phenyl O t , ,t~ CO 2~rboxy-6- CO H 1 3 benzene hydroxy phenyl 109 NH NH C0 o-chloro phenyl O methoxy CO 2-carboAy-6- CO H 1 3 c benzene methoxy phenyl ~O
110 NH NH C0 p-methoxy phenyl O :' huAy CO 2-hydroxy-6- CO H 1 3 jR
benzene methyl phenyl . ~
Cmpd. ¦ A E~ E~ D E F G X K m n ¦ ~ ~ g 111 NH NH CO p-chloro phenyi O t~lr.. hoAy CO 2,6-dimethyi CO H 1 3 benzene phenyi 112 NH NH CO p-hydroxy phenyi O tetramethyi CO 2,6-dihydroxy CO H 1 3 benzene phenyi 113 NH NH CO p-rnethoxy phenyi O ethoxybenzene CO 2-carboxy4- CO H 1 3 methyi benzene 114 NH NH CO p-hydroxy phenyi O diethoxy CO 2-carboxy4- CO H 1 3 benzene hydroxy phenyi 115 NH NH CO m-hydroxy phenyi O nitrobenzene CO 2-carboxy4- CO H 1 3 methoxy phenyl 116 NH NH CO p-chloro phenyi O dinitrobenzene CO 2,6-dimethyi CO H 1 3 phenyi 117 NH NH CO o-methoxy phenyi O halobenzene CO 2-hydroxy4- CO H 1 3 1.
methyi phenyi O
118 NH NH CO p-methoxy phenyi O dihalobenzene CO 2-carboxy4- CO H 1 3 methyi phenyi 119 NH NH CO p-hydroxy phenyi O trihalobenzene CO 2-hydroxy4- CO H 1 3 methyi phenyi 120 NH NH CO o-chloro phenyi O tetrahalo CO 2-carboxy4- CO H 1 3 benzene methoxy phenyi 121 NH NH CO m-hydroxy phenyi O carboxyacid CO 2-carboxy4- CO H 1 3 benzene hydroxy phenyi 122 NH NH CO m-chloro phenyi O ' bOAj ld CO 2-carboxy4- CO H 1 3 c benzene ethyi benzene v 123 NH NH CO p-hydroxy phenyi O amido benzene CO 2,6-di".~t'loiy CO H 1 3 ,~
phenyi 00 124 NH NH CO o-methoxy phenyi O diamido CO 2-c~rboxy4- CO H 1 3 benzene hydroxy phenyi -Cmpd. A¦ B1 ¦ B2 ¦ Z D E F G ¦ X K m n ¦ o 125 NH NH CO p-hydroxy phenyi O phenol CO 2-carboxy4- CO H 1 3 hydroxy phenyl 126 NH NH CO 1.. hùAy phenyl O dihydroxy CO 2-; LUAY6- CO H 1 3 benzene hydroAvy phenyi 127 NH NH CO p-methoAy phenyi O trihydroAy CO 2-carboxy4- CO H 1 3 benzene propoxy benzene 128 NH NH CO 1.. 3~ UA~ phenyl O l~,t~ hJJ~UAy CO 2carboxy4- CO H 1 3 benzene hydroxy phenyi 129 NH NH CO p-chloro phenyi O triethoxy CO 2,6-dimethyi CO H 1 3 benzene phenyi 130 NH NH CO p-methoAy phenyi O tetraethoxy CO 2-carboxy4- CO H 1 3 benzene methoxy phenyi I
131 NH NH CO p-hydroxy phenyi O pentoAy CO 2,6-dihydroxy CO H 1 3 1~ C r~
benzene phenyl ~ _~
132 NH NH CO p-hydroxy phenyi O dipentoxy CO 2,6~ . t~10AY CO H 1 3 benzene phenyl 133 NH NH CO m-methoAy phenyl O tripentoAvy CO 2- buAy6- CO H 1 3 benzene ethyi benzene 134 NH NH CO o-hydroxy phenyi O I ~r~ Y CO 2-carboxy4- CO H 1 3 benzene . hydroxy phenyi 135 NH NH CO o-hydroxy phenyi O aniline CO 2-carboxy4- CO H 1 3 hydroxy phenyi 136 NH NH CO o-chloro phenyi O dbmino CO 2,6-~ thoAy CO H 1 3 C
benzene phenyi ~, 137 NH NH CO p-methoAy phenyi O methoxy CO 2-carboxy4- CO H 1 3 pyridine propyi benzene A
2157~
E
_ y I
O O O O O O O O O O O O O
N E N N N E N ~ N N N E N N ~D N _ N E N
O O O O O O O O O O O O O
~ C ~ t ~ 2 E
C~
O O O O O O O O O O O O O
~ ~ C G C ~ lii c ~ CL ~ t~. G
G G ~ O O O
m O O O O O O O O O O O O O
m Z Z Z Z Z Z Z Z Z Z Z Z Z
I
Z Z Z Z Z Z Z Z Z Z Z Z Z
c E 0 0 -- N ~ ~ It~ O
e~ e~ e~ e~ e!~ e~ e!~ e~ e~ eq e!~ e~ e~
E _ _ _ _ _ _ _ _ _ _ _ _ _ Y
- O O O O O O O O O O O O O
~ '~
e~ e~
'. '. eD~U ~U 'U
N . N . G C~ O .E ~ 2 : L
O O O O O O O O O O O O O
-2 2 3~ 3O~ ~ 3O~
LU ,. eD ~ g~ U g, ~ U ~ l U ~ I
e~ Ce!~ _ e!~ _ e!~ _er~ _ e!~ . eq . e~ _ e!~ . eq . e!~ _ eq .
O O OO O O O O OO O O O
~U ~ eD eu eu eeu ~ eu e ,eu eu eu O OG CL ~ O C~
m O OO O O O O OO O O O O
~ o m Z Z Zz z z z z zZ Z z z I
Z Z Zz z z z Z Zz Z z z c u~ eD r~ e~ u~~OD ~ ~ eD
C~ ~ _ __ _ _ _ _ __ _ _ _ _ Cmpd. ¦ A B1 B2 Z D E F G X K m n ¦ O
166 NH NH CO p4hloro phenyl 0 3,5-dihydroxy CO thbzole CO H 1 3 benzene 167 NH NH CO m-methoAy phenyl 0 3,5-dihydroxy CO Isoxazole CO H 1 3 benzene 168 NH NH CO p-chloro phenyl 0 3,5-dihydroxy CO pyrazole CO H 1 3 benzene 169 NH NH CO p-hydroxy phenyl O benzene isutl,' '~ CO H 1 3 170 NH NH CO p-methoAy phenyl 0 3,5-dihydroxy CO benzene CO H 1 3 benzene 171 NH NH CO o-hydroxy phenyi 0 3,5-dihydroxy CO methyibenzene CO H 1 3 benzene 172 NH NH CO o-chloro phenyl 0 3,5-dihydroxy CO dimethytbenzene CO H 1 3 ~' benzene ,~
173 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO trimethylbenzene CO H 1 3 benzene 174 NH NH CO p-me~hoxy phenyi 0 3,5-dihydroxy CO t~ tl,~l CO H 1 3 benzene 175 NH NH CO ,.. n lhoAy phenyl 0 3,5~ihydroxy CO ethylbenzene CO H 1 3 benzene 176 NH NH CO p-hydroxy phenyl 0 3,5-dihydro~ CO tetraethylbenzene CO H 1 3 benzene 177 NH NH CO m-hydroxy phenyt 0 3,5-dihydroxy CO propyibenzene CO H 1 3 c benzene benzene u 173 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO tetrapropyi CO H 1 3 benzene benzene 179 NH NH CO p-chloro phenyl 0 3,5-dihydroxy CO butylbenzene CO H 1 3 _ Y
- O O O O O O O O O O O O O
~ "
~ n ~ ~. .. ~
~ , . C . ~ . ~ _ o o o o o o o o o o o o o o J~ ' ; ~ ~L ~ o ~ O ~ o2 2 o o o o o o o o o o o o o ,c C c~ C~ C C~ c 1~ ~ Q, G 'c~ ~
c~ ~ G ~ c~ ~. G ~ G
,o ,o ,o C- ~ O O G CL G i o m I
E 0 0 '`~ D ~ 0 ~ o, Cmpd. A ¦ B~ B2 Z D E F G X K m n ~ 2 193 NH NH CO ... h~J~UAy phenyi 0 3,5-dihydroAy CO trihalobenzene CO H 1 3 benzene 194 NH NH CO m-chloro phenyi 0 3,5-dihydroAy CO tetrahalobenzene CO H 1 3 benzene 195 NH NH CO p-hydroxy phenyi 0 3,5"hyJ,JA~ CO carboxyadd CO H 1 3 benzene benzene 196 NH NH CO p-chloro phenyi 0 3,5-dihydroAy CO " ~ ~,f ld CO H 1 3 benzene benzene 197 NH NH CO p-methoAy phenyi 0 3,5-dihydroxy CO amido benzene CO H 1 3 benzene 198 NH NH CO m-hydroA~y phenyi 0 3,5-dihydroAy CO diamido benzene CO H 1 3 benzene 199 NH NH CO m-chloro phenyi 0 3,5-dihydroxy CO 3,5-dihydroxy CO H 1 3 1' benzene benzene a~
200 NH NH CO o-methoAy phenyi 0 3,5-dihydroA~y CO trihydroxybenzene CO H 1 3 benzene 202 NH NH CO o-methoAy phenyi 0 3,5-dihydroA~y CO t `r~hJJ~uAy CO H 1 3 benzene benzene 202 NH NH CO p-chloro phenyi 0 3,5-dihydroAy CO p~.~' hJJ~uA~ CO H 1 3 benzene benzene 203 NH NH CO p-chloro phenyi 0 3,5-dihydroAy CO triethoAybenzene CO H 1 3 benzene 204 NH NH CO o-hydroAy phenyi 0 3,5-dihydroxy CO tetraethoAy CO H 1 3 c benzene benzene r~
205 NH NH CO p-hydroAy phenyi 0 3,5-dihydroAy CO pentoxybenzene CO H 1 3 benzene ~, 206 NH NH CO p-hydroxy phenyi 0 3,5-dihydroxy CO dipentoxybenzene CO H 1 3 benzene Cmpd. A B~ ¦ J2 ¦ ¦ E F G X K m n 207 NH NH CO p-methoAy phenyl 0 3,5~ihydroxy CO tripen~oxybenzene CO H 1 3 benzene 208 NH NH CO m-hydroxy phenyl 0 3,5-dihydroxy CO l~t,., , y CO H 1 3 benzene benzene 209 NH NH CO o-methoAy phenyi 0 3,5-dihydroxy CO amino benzene CO H 1 3 benzene 210 NH NH CO o-chloro phenyi 0 3,5-dihydroxy CO diamino benzene CO H 1 3 benzene 211 NH NH CO p-methoxy phenyi 0 3,5-dihydroxy CO methoxypyridine CO H 1 3 benzene 212 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO Ji",_lhuA~pyridine CO H 1 3 benzene I ~V
213 NH NH CO p-methoxy phenyi 0 3,5-dihydroxy CO hydroAypyridine CO H 1 3 benzene 214 NH NH CO p-methoxy phenyl 0 3,5-dihydroxy CO dihydroxypyridine CO H 1 3 1 ~_~
benzene l~
215 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO ethoAypyrrole CO H 1 3 benzene 216 NH NH CO ... hJJ~uAy phenyi 0 3,5-dihydroxy CO dihydroxypyrrole CO H 1 3 benzene 217 NH NH CO p-chloro phenyi 0 3,5-dihydroxy CO ' . `i~.. A~Hndole CO H 1 3 benzene 218 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO hydroAypurine CO H 1 3 c benzene u 219 NH NH CO p-methoxy phenyi 0 3,5-dihydroxy CO d~.,._lhuAyfuran CO H 1 3 benzene 220 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO hydroxyll, ~ .,t CO H 1 3 benzene ~ o Cmpd. A ~1¦ i32 Z D E F G X K m n Wg 221 NH NH C0 m-chloro phenyi O benzene CO methoAyl".; ' ,e CO H 1 3 222 NH NH C0 p-hydroAy phenyi 0 3,5-dihydroAy CO 'i.. IhuAy CO H 1 3 benzene IJ j . ;~;. ,e 223 NH NH C0 p-chloro phenyl O benzene CO hydroAyp,.; " ,e CO H 1 3 224 NH NH C0 m-chloro phenyi 0 3,5-dihydroAy CO diamido py.; .;di,.e CO H 1 3 benzene 225 NH NH C0 o-hydroAy phenyi 0 3,5-dihydroxy CO amido pyrazine CO H 1 3 benzene 226 NH NH C0 m-hydroAy phenyl 0 3,5-dihydroA~y CO diethoAypyrazine CO H 1 3 benzene 227 NH NH C0 quinoline 0 3,5-dihydroAy CO phenyi CO H 1 3 1--benzene 22a NH NH C0 methoxy 0 3,5-dihydroxy C0 phenyi CO H 1 3 quinoline benzene 229 NH NH C0 ~'. hùAy 0 3,5-dihydroAy C0 phen~ CO H 1 3 quinoline benzene 230 NH NH C0 I,i hUA~ O 3,5-dihydroAy CO phenyl CO H 1 3 quinoline benzene 231 NH NH C0 hydroAy 0 3,5-dihydroxy C0 phenyi CO H 1 3 quinoline benzene 232 NH NH C0 dihydroAy 0 3,5-dihydroAy C0 phenyl CO H 1 3 c quinoline benzene u 233 NH NH C0 ethoAy 0 3,5-dihydroxy C0 phenyl CO H 1 3 ~;
quinoline benzene 234 NH NH C0 amino 0 3,5-dihydroAy CO phen~ CO H 1 3 quinoline benzene Cmpd. A 31 B2 7 D E ¦ F G ¦ X ¦ K m n 235 NH NH CO dbmido 0 3,5-dihydroxy CO phenyi CO H 1 3 qulnoline benzene 236 NH NH CO trihalo 0 3,5-dihydroAy CO phenyi CO H 1 3 quinoline benzene 237 NH NH CO quinolinecarboA~yic O 3,5-dihydroAy CO phenyi CO H 1 3 acid benzene 238 NH NH CO ~,ln l.e O 3,5-dihydroAy CO phenyi CO H 1 3 benzene 239 NH NH CO methoAy O 3,5-dihydroAy CO phenyi CO H 1 3 ~il ' , " ,e benzene 240 NH NH CO . ~lh~AY O 3,5-dihydroAy CO phenyi CO H 1 3 'il' . ' ,e benzene 241 NH NH CO l~i\, Ihv~ O 3,5-dihydroAy CO phenyi CO H 1 3 ~ r~
il' ,- ',e benzene ~D
242 NH NH CO hydroxy O 3.5-dihydroAy CO phenyi CO H 1 3 1 _~
, ', ' ,e benzene ~p.
243 NH NH CO t~;hy.' VAY O 3,5-dihydroxy CO phenyi CO H 1 3 ~,-ln !ine benzene 244 NH NH CO t hv~y O 3,5-dihydroAy CO phenyi CO H 1 3 e benzene 245 NH NH CO diamino 0 3,5-dihydroxy CO phenyi CO H 1 3 il~ln ' ,e benzene 246 NH NH CO triamido 0 3,5-dihydroAy CO phenyi CO H 1 3 q~lr ' ,e benzene ~
247 NH NH CO tetrahalo 0 3,5-dihydroAy CO phenyi CO H 1 3 O
, ', 'i. ~e benzene 248 NH NH CO 'i"~ ~'h,e O 3,5-dihydroxy CO phenyi CO H 1 3 Ji~_. i VAyi- acid benzene 21S7~ 120-E _ _ _ _ _ _ _ _ _ _ _ _ _ _ O O O O O O O O O O O O O O
C C C C ~ &
G 1~ C~ ~ G C~ C~ ~ C~ 1~ D ~ C~ C~
LL
ID
o ~ ~ O ~
O O O O O O O O O O O O O O
C ~ ~ C C ~D ~ G C~ ~ ~ G
: : : . : :
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m O O O O O O O O O O O O O O
m Z Z Z Z Z Z Z Z Z Z Z Z Z Z
ZZZZZZZZZZZZZZ
-E ~ O -- N 0 ~ U) ~ ~-- 0 CO O _ N
~le N N N N N N N N N N N N N N
WO 94/20062 215 7 ~ 12 PCT/US94/02283 E _ _ _ _ _ _ _ _ _ _ _ _ _ Y
x 8 ~, ~, ~, ~, 8 ~, ~, ~ ~
G G C~ Cl. ~ G ~ CL ' ' C
O O O O O O O O O O O O O
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N
m O O O O O O O O O O O O O
m Z Z Z Z Z Z Z Z Z Z Z Z Z
-T T I I I T I I ~
Z Z Z Z Z Z Z Z Z Z Z Z Z
~
C.) N N N N N NN N N ~N` N C~l N
WO 94/20062 PCTtUS94/02283 21~7412 - 122 -E
Y
O O O O O O O O O O O O O O
2 ~ o ~ o : : o ~ .E
8 8 8 ~, 8 8 8 8 8 8 8 8 8 8 2 ,~ D ~ ~ C ~ S
O O O O O O O O O O O O O O
G 0- G ~ ~ CL , ~ ~ ' ~ c G
c c~ G ~ c o o o `.
m O O O O O O O O O O O O O O
m ZzzzzzzzzzzzzZ
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E ~D ~ 0 0 c~ _ N C" ~ U~ ~ ~ 0 0 C~ ~1. N N N N N0 N0 N0 N N0 N0 N0 N N N0 WO 94120062 21~ 7 412 PCTIUS94/02283 E _ _ _ _ _ _ __ _ _ _ _ Y
~ ~ ,,. . ~
_ 8 8 8 o 8 8 8 8 8 8 8 8 8 o o o o o o o o o o o o o G ,c ~ ~ ~ G G ,c . . O : ' G ~ . C G O
m O O O O O O O O O O O O O
m I I I I I I I I I I
Z Z Z O O O ~ Z c~ ~o Z Z Z Z Z
~ ' N N N N 0 N 0 r-- 0 0 0-- N
cn ~`~
Cmpd. A B 1 B 2 Z D E F G X K m n f 303 NH O CO m-hydroxy phenyi O 3,Wihydroxy CO phenyi CO H 1 3 benzene 304 NH N-ethyi CO o-chloro phenyi O 3,5-dihydroAy CO phenyi CO H 1 3 benzene 305 NH N- CO ... mlhuAy phenyi O 3,5-dihydroxy CO phenyi CO H 1 3 methyi benzene 306 N-anisly N- CO p-hydroxy phenyi O 3,5-dihydroAy CO ;l~ln 'ine CO H 1 3 phenyi benzene " bGAJr': acid 307 NH NH CS p-chloro phenyi O 3,5 :"hJ ' UAy CO phenyi CO H 1 3 benzene 308 NH NH CO p-hydroxy phenyi CH2 3,5-dihydroAy CO phenyi CO H 1 3 benzene 309 NH NH CO p-hydroxy phenyi N-ethyi 3,5-dihydroAy CO phenyi CO H 1 3 N
310 NH NH CO p-methoxy phenyi N- 3,5-dihydroAy CO phenyi CO H 1 3 phenyi benzene 311 NH NH CO o-methoxy phenyi N- 3,5-dihydroxy CO phenyi CO H 1 3 propyi benzene 312 NH NH CO m-chloro phenyi O 3,5-dihydroxy CS phenyi CO H 1 3 benzene 313 NH NH CO m-chloro phenyi O 3,5-dihydroxy CH(OC phenyi CO H 1 3 benzene H ~ ~, 314 NH NH CO o-chloro phenyi O 3,5-dihydroAy CH(O- phenyi CO H 1 3 benzene phenyi) u 315 NH NH CO o-hydroxy phenyi O benzene CH2 phenyi CO H 1 3 O
316 NH NH CO m-chloro phenyi O 3,WihydroAy O phenyi CO H 1 3 benzene WO 94/20062 215 7 ~ 1~ PCT/US94/02283 r.~ -- r,~l ~ . _ _ N N r.~ r,~ _ _ N
-- _ _ _ N r~ ~ N e~ N
Y I I I I I T
8 8 8 8 8 8 8 8 8 8 8 8 ~ 8 ~
~ ;~, f ~ f f f f f ~ ~D f f ~~D
~ 8 8 8 8 8 8 8 8 8 8 ~ 8 ~ 8 ~i b ~ C C ~ D
~ . ~ . r, ~, r,.~, r, ~ rq _ ~ _ ~ . ~ . _ . C~ . .
O O O O O O O O O O O O O O O
c c ~ ~ ~ ~ ~ _ ~
~1 ~ ~ ~, ~ ~ ~. ~ ~, ~ rD
G G C~ i O C~ G 2 . : : .
_ 8 8 8 8 8 8 8 8 8 8 8 o 8 ~
m I I I I T
Z Z Z z z z z z z z a~ Z o. Z Z Z
E ~ N N CNU N N N N N N N c~ C~
2157 ~ 126 -N N _N _ 0 0 0 0 _ N 0 E __ _ _ _ _ _ _ _ _ _ _ N
Y II I I I I T I I I I T
Otn Ou~ O a~ O u~ Z, ,~ ~ O O
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O O O O O o~ O a~ Ou~ O c/~ O a~
D ~ D 'D _ E _ 2 ~
~ . . . ~
0~ O~n O O O O U~ I _ I ~ O O O
O O U~ C O ~
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WO 94t20062 PCTtUS94/02283 2157ll~2 N ~t _ _ _ N N c~ _ _ N
E _ _ N ~ ~ N C'~ N
Y
O a~ O a~ Z~,~U O O o o cn O O O
'U
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m t~ O a~ z '' z E ' ~ ' ~
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Za~Z~ZZZZOt~ZZZZ
c-- Co _ N 7 ~ In tD
WO 94/20062 215 7 ~12 PCT/US94102283 N ~ N ~ _ _ E _ _ _ _ _ _ _ _ _ _ _ N C'~
I T I I I I I T T T I I T
X N
O O O tn O o~ O a~ O o~ o 0, z C~ ~ ,'D ~ o ~ ; ~ ; ; ; , O O O O O cn O O a~ O
O Z G ~1) ~ 2 ~ 2 ~ o ' G ' Q ~ Q ~ ~ b . ~ . ~ . ~ . ~ . ~ ~ c -o o o o o o o o o o o o o G Q Q ~Q 'G C ~ D C ~ ~D
N ~ ~ Q ~ ~ ~, ,~
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N
m O O O o O~n O O ~ O O O O
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386 CH2 NH CS o-methoxy phenyi O methoxy CS amino benzene CS H 4 e 387 0 NH CO o-hydroxy phenyi O h 'hJAY CO diamino benzene CH(CH3 H 2 2 ~".; ' ' .e CH3~
388 S NH CO ... n, Iho,~y phenyi O hydroxy N- methoxypyridine CS H 3 2 .. IJi"e methyi 389 N-SO z NH CO p-chloro phenyi NH dbmido CO d,,,,~lhùxypyridine C=N- H 2 3 phenyi 1,,.; .. Ji"e phenyi 390 N-methy NH CO p-chloro phenyi O amido pyrazine N- hydroxypyridine CS H 1 3 pyridine 391 N-butyi NH CO p-hydroxy phenyi NH diethoxy CO dihydroxypyridine CO H 1 3 pyrazine 392 N-phenyi NH CO p-chloro phenyi NH phenyi N- phenyi CO H 1 3 1' purine o 393 S NH CO p-hydroxy phenyi NH phenyi CO phenyi CO H
394 N-SO z NH CS hydroxytl,', h~ne NH dihydroxy CH2 phenyi CO H
phenyi benzene 395 N-methyi NH CO methoxy~,.'' '.e CH2 dihydroxy CH2 phenyi CO H 1 4 benzene 396 N-butyl NH CS ~" . 'huAy O dihydroxy O phenyi CO H 2 ~,.;JaL;"e benzene 397 N-phenyi CH2 CO hydroxypf~ ' " ,e O dihydroxy O phenyi CO H 3 1 ~
benzene 6 398 N-pyridy O CS diamido ~".; ,;di"e N- dihydroxy S pheny CS H 4 1 phenyi benzene 399 0 0 CO amido pyrazine N- dihydroxy CH2 phenyi CO H 2 2 methyi benzene WO 94/20062 21~ 7 ~12 PCT/US94/02283 ._ N ~ ~r _ _ N _ N N _ N _ C'7 E ,., N
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benzene methyi benzene 457 N-phenyl NH CO pentoxy benzene O phenyl CO benzene CO H 1 2 458 N-pyridyi NH CS dipentoxy benzene O phenyi N-ethyi dihydroxy CO H 1 4 benzene 459 NH NH CO tripentoxy benzene O phenyi CO dihydroxy CS H 2 benzene 460 S NH CO l-,t~ O phenyi N- dihydroxy CO H 3 1 1' ~3 benzene purine benzene Ul t~
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rD ~ r, WO94/2~K2 PCT~S94/0~83 21~ ~12 The following, nonlimiting Examples illustrate preferred methods for preparing the compounds for use in the method of the present invention and the data demonstrating PKC
inhibitory activity of the Compounds used in the present invention.
EXAMPL~8 3-Acetyl~minohexahydro-l-phenylmethylazepin-2,~-dione A solution of hexahydro-l-phenylazepin-2,3,4-trione-3-oxime (1.23 g, 5mmol) in 4:1 acetic acid/acetic anhydride (20 ml) was treated with Raney nickel (Aldrich, one-half tsp) in a Parr bottle and subjected to hydrogenation for eighteen hours at 40-45 psi and room temperature. The mixture was carefully evacuated of hydrogen and filtered through Celite~. The filter pad was then washed with methanol (with care taken not to let the filter pad become dry). The filtrate was concentrated in vacuo and the residue was diluted with toluene and further concentrated to remove most of the acetic acid. The residue was chilled on an ice bath and treated with saturated sodium bicarbonate carefully to avoid excessive bubbling. The cloudy aqueous solution was extracted with methylene chloride (3 x 50 ml), and the combined organic solution was dried (Na2SO4) and concentrated in vacuo. The residue was flash chromatographed on silica gel (eluted with 19:1 methylene chloride/methanol) to afford 3-acetylaminohexahydro-1-phenylmethylazepin-2,4-dione (1.11 g, 81~) as a white solid.
syn-3-Aminohexahy~ro-~-hy~roxy-l-phenylmethylazepin-2-one A solution of 3-acetylaminohexahydro-1-phenylmethylazepin-2,4-dione (0.82 g, 3.0 mmol) in absolute ethanol (lS ml) was treated with sodium borohydride (0.23 g, 6 mmol) and stirred for thirty minutes. The solution was then treated with water (5 ml) and concentrated in vacuo, and taken up in 2:1 ethanol/water (7.5 ml). Concentrated hydrochloric acid (2.5 ml) was added, and the mixture was refluxed for two hours and partially concentrated, then diluted with water (25 ml). The aqueous acidic mixture was extracted with ether (25 W094l20062 PCT~S94/02283 ml), and the aqueous solution was basified with 30% sodium hydroxide and extracted with methylene chloride (3 x 40 ml).
The combined methylene chloride extracts were washed with water (25 ml), dried (Na2S04), and concentrated in vacuo, to a yellow solid, which was recrystallized from ethyl acetate to afford syn-3-aminohPYAhydro-4-hydroxy-1-phenylmethylazepin-2-one (0.42 - g, 60%) as a white solid.
syn-3-Aminohexahydro-~,-hydroxy-l-phQnylmethylazepine A cooled (S-C) solution of lithium aluminum hydride/tetrahydrofuran (Aldrich, 1Ø N, 5.1 ml) under nitrogen was treated with syn-3-aminohexahydro-4-hydroxy-1-phenlmethylazepin-2-one (0.40 g, 1.7 mmol) in portions so that the pot temperature did not exceed 15-C. The mixture was refluxed for 6.5 hours, cooled on an ice bath, and carefully treated with water (0.21 ml), 15% sodium hydroxide (0.21 ml), and water (0.63 ml). The suspension was allowed to stir for five days, during which time the product partially decomposed (optimal time is 2-5 hours). The suspension was filtered, and the filtrate was concentrated in vacuo and chromatographed on silica gel (eluted with 90:8:2 methylene chloride/methanol/triethylamine). The appropriate fractions were concentrated in vacuo to afford syn-3-aminohexahydro-4-hydroxy-1-phenyl-methlazepine (0.22 g, 58%%) as a colorless oil.
syn-~exahydro-~-hydroxy-3-(~-phenylmethoxy)benzoyl~mino-1-phenylmethyl~zepine A solution of 4-benzyloxybenzoic acid (0.183 g, 0.8 mmol) in anhydrous tetrahydrofuran (2 ml) and N, N-dimethylformamide (0.5 ml) was treated with N,N'-carbonyldiimidazole ((0.15 g, 0.9 mmol) and stirred at roomtemperature for l.Sh. The solution was treated with syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.20 g, 0.9 mmol) in anhydrous tetrahydrofuran (1 ml). The mixture was stirred for eighteen hours, and then concentrated in vacuo.
the residue was taken up in lN sodium carbonate (20 ml), and W094l2~62 PCT~S94/02283 21~7~12 - 162 -the aqueous mixture was extracted with toluene (2 X 25 ml) containing a small amount of 2-propanol. The combined organic extracts were dried (Na2SO4) and the concentrated residue was flash chromatographed on silica gel (eluted with 3:1 ethyl acetate/hexane) to afford syn-hexahydro-4-hydroxy-3-(4-penylmethoxy)-benzoylamino-l-phenylmethylazepine (0.17 g, 50%) as a viscous oil.
3,5-Bis~phenylmethoxy)-~-~2-phenylmethoxybenzoyl benzoic ~ci~
ester with syn-Hexahy~ro-~-hydroxy-3-~4-phenylmethoxy benzoyl-~mino-l-phenylmethylazepine A solution of 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)benzoic acid (0.245 g, 0.45 mmol) in anhydrous methylene chloride (1.5 ml) was treated with N,N-dimethylformamide (two drops), then with 2 N oxalyl chloride/methylene chloride (Aldrich, 0.30 ml, 0.60 mmol), and stirred for one hour under nitrogen. The solution was concentrated in vacuo and placed under high vacuum for one hour. syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoyl-amino-l-phenylmethylazepine (0.170 g, 0.40 mmol) was dissolved in anhydrous methylene chloride (3 ml), treated with 4-dimethyl-aminopyridine (0.01 g) and triethylamine (0.12 ml, 1.2 mmol), and cooled in an ice bath under nitrogen. The acid chloride was removed from high vacuum and dissolved in anhydrous methylene chloride (2 ml), and was then added to the cooled solution. The mixture was allowed to warm to room temperature, stirred for one hour, and was partially concentrated in vacuo. The residual solution was deposited on a silica gel column and eluted (first with 2:1 hexane/methylene chloride, then with 1:1 hexane/methylene chloride) to afford (after concentration of the appropriate fractions) 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)- benzoic acid ester with syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepine (0.29 g 77~) as a white foam.
N O 94/20062 21~ 7 412 PCTrUS94tO2283 syn- (~- (3,5-Dihydroxy-~-(2-h~.osybenzoyl)benzoyloxy))hexhy~ro -3-(~-hydroxybenzoylamino)azepine A cloudy suspension of 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl-benzoic acid ester with syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepine (0.29 g, 0.30 mmol) in reagent ethanol (25 ml) was treated with - Pd(OH)2/C (Aldrich, Pearlman's catalyst, 0.20 g, 20%) in a Parr bottle, then subjected to hydrogenation for twenty four hours at 50-52 psi. The hydrogen was evacuated and the solution was carefully filtered through Celite~ under nitrogen, and the filter pad was washed with methanol (not to dryness). The filtrate was concentrated in ~acuo to crude material, which was flash chromatographed on a short column of silica gel (eluted with 1:1 CHClJEtOH) to afford a product (0.13 g) as a pale yellow foam. This was triturated from ether/ acetonitrile to afford syn-(4-(3,5-dihydroxy-4-(2-hydroxybenzoyl-benzoyl-oxy))hexahydro-3-(4-hydroxybenzoylamino) azepine (0.195 g, 62%) as a pale yellow powder (dihydrate): mp 177-179-C; Rf (1:1 CHCl3/EtOH on silica) 0.45; IR (KBr) 1623 cm , lH NMR (d6-DMSO) ~ 8.20 (d, lH, J = 8Hz), 7.10 (s, 2H), 7.02 (d, lH, J - 9 Hz), 6.91 (t, lH, J -= 8Hz), 6.78 (d, 2H, J = 9Hz), 5.39 (br d, lH, J = 7Hz), 4.48 (m, lH), 3.00-3.20 (m, 4H), 2.05-2.20 (M, lH), 1.70-2.00 (m, 3H). Anal. Calcd. for C27H26N2O8-2H20: C, 59.77; N,
5.57; H, 5.16. Found: C, 59.83; H, 5.39; N, 5.46.
anti-4-~3,5-Dihydroxy-~-~2-phenylcarbonyl-benzoyloxy-3-(~-hydroxybenzamido)azepine To a solution of tran-4-(3,5-dibenzyloxy-4-phenyl carbonyl-benzoyloxy-3-(4-benzyloxybenzamido)-N-benzylazepine (40 mg, 0.042 mmol) was added Pd(OH)2 (Pearlman's catalyst) 20 mg, 50% on weight basis) and introduced H2 gas at atmosphere pressure.
Trans-l-(~-hydroxybenzamido)-2-~4-benzyl-3,5-dihydroxybenzoyloxy) cycloheptane The catalyst Pd(OH)2 on carbon (20%, moist, 20 mg) was added to a solution of trans-1-(4-benzoyloxybenzamido)-2-)4-W094l2~62 PCT~S94/02283 21S7~1Z - 164 -benzoyl-3,5-dibenzyloxybenzoyloxy)cycloheptane (215 mg, 0.28 mmol) in methanol (8.4 ml). The mixture was stirred vigorously at room temperature under 1 atm H2 contained in a balloon for sixteen hours. The solid catalyst was removed by flash chromatography (sioz~ 2:2:1/diethyl ether:hexane:methylene chloride) to give a white powder (112 mg, 84%): mp 224-226-C;
lH NMR (CD30D) ~ 7.52-7.56 (m, 2H), 7.02-722 (m, 5H), 6.94 (s, 2H), 6.71-6.75 (m, 2H), 5.10 (tm, J = 9.1 Hz, lH), 4.35 (tm, J-9.3 Hz, lH`), 3.93 (s, 2H), 1.56-2.02 (m, lOH) IR (KBr) cm 3389, 1687, 1626. Anal. calcd. for C28H29O6N: C, 70.72: H, 6.15:
N, 2.95. Found C, 70.39; H, 6.37; N, 2.67.
Trans~ -hydroxybenzamido)-2-[~-(2-hydroxybenzoyl)-3,5-dihy~roxybenzoyloxy] cycloheptane The catalyst Pd(OH)2 on carbon (20%, moist, 9 mg) was added to a solution of trans-1-(4-benzyloxybenzamido)-2-t4-(2-benzyloxybenzoyl)-3,5-dibenzyloxybenzoyloxy~ cycloheptane (112 mg, 0.13 mmol) in methanol (3.9 ml) and ethyl acetate (1.3 ml).
The mixture was stirred vigorously at room temperature under 1 atm H2 contained in a balloon for seventeen hours. The solid catalyst was removed by filtration through Florisil. The filtrate was evaporated and purified by flash chromatography (SiO2 2:2:1/ethyl acetate:hexane:methylene chloride) to give a pale yellow powder (40 mg, 61~): mp 234-236-C; lH NMR (CD30D) ~ 7.56-7.59 (m, 2H), 7.47 (t, J-7.1 Hz, lH), 7.23 (d, J=8.0 Hz, lH), 7.00 (s, 2H), 6.96 (d, J=8.2 Hz, lH), 6.74-6.78 (m, 2H), 5.15 (tm, J=9.3 Hz, lH), 4.40 (tm, J=9.3 Hz, lH), 1.58-2.05 (m, lOH) IR (KBr) cm 3392, 1700, 1678, 1626. Anal. calcd. for C28H29O6N: C, 66.53; H, 5.38 N, 2.77. Found: C, 66.37; H, 5.56; N, 2.47.
wos4/20062 _ 165215 7 4 12 PCT~S94/02283 (+)-Tr~ns-4-[~-(2-Ca.LGa~ 6-hy~roxybenzoyl)-3,5- ~ihy~roxy-benzoyloxy]-3-~-hydroxybenzami~o)-1-~methylsulfonyl) pyrrolidine ~COMPO~ND 501) HO N31 H2' 10h Pd/C HO~ ~N~
~j CH30 H , ~ ~OBn N 2. THF, 2 N KOH N
- S 02CH3ClOC~OBn S 02CH3 C 02Bn NE~, l o OBn DMAP, C3~COCI
OBn HO~ N~3~ EP~dOOHHT2FA BnO~O NJ~3 h OH 0~ h OBn H SO2CH3 BnO2C~b 8BBn N
~+)-Tr~ns-~-~ydroxy-3-~-benzylGxyLr~z~mido)-l-~methylsulfonyl) -pyrrol~dine To 10% palladium on carbon (41 mg) wetted with methanol (1.0 ml) was added the azide (412 mg, 2.00 mmol) in methanol (9.0 ml). The flask was evacuated and filled with H2 twice then allowed to stir under H2 (1 atm) for 4 h. The mixture was filtered through Celite and washed through with methanol (100 ml). The methanol was evaporated, providing an off-white solid which was used without characterization.
To a O C solution of the above amino alcohol in THF
(6.0 ml) was added 2 N KOH (1.0 ml). The ice bath was removed, and the acid chloride (approx. 0.75 eq) added portionwise over 2.5 h, until the starting material was gone as evidenced by thin layer chromatography. The mixture was diluted with CH2Cl2 (30 ml) and poured into H2O (60 ml). Methanol (20 ml) was W094/2~62 PCT~S94/02283 21S7~2 - 166 -added to disperse the emulsion, the layers were separa~ed, and the aqueous layer extracted with CH2C12 (4 x 50 ml). The organic lyaers were combined, dried (MgSO4) filtered and evaporated to provide the title compound as a white solid (593 mg, 76% over two steps).lH NMR (CD30D) ~ 7.60 (d, J = 8.7 Hz, 2H), 7.25-7.10 (m, 5H), 6.85 (d, J = 9.0 Hz, 2H), 4.94 (s, 2H), 4.18-4.12 (m, 2H), 3.58-3.42 (m, 4H), 2.71 (s, 3H).
(~)-Tr~ns-~-[~-~6-benzyloxy-2-(benzyloxycarbonyl)benzoyl]-3,5-~ibenzyloxybenzoyloxy)-3-(~-~enzyloxybenzami~o)-1-~methyl sulfonyl)-pyrrolidi~e ~COMPOUND 600) To a 0-C solution of the product of the previous step (202 mg, 0.518 mmol), diisopropylethylamine (98 ~1, 1.2 eq., 0.565 mmol) and 4-dimethylaminopyridine (58 mg, 1.0 eq., 0.471 mmol) in CH2Cl2 (8.0 ml) under N2 was added a solution of the acid chloride (0.471 mmol) in CH2C12 (4.0 ml). The reaction was allowed to warm to room temperature with stirring over 16 h.
The cloudy reaction mixture was diluted with CH2Cl2 (50 ml) and washed with satd. NaHCO3 (30 ml) then brine (30 ml). The aqueous layers were extracted with CH2C12 (50 ml) each. The organics were combined, dried (MgSO4), filtered and evaporated to a light yellow oil. Flash column chromatography (1:1 hexane:ethyl acetate) provided the title product as an off white foam (371 mg, 75%): mp 73-79-C; IR (KBr) 3367, 1723, 1660, 1230, 1112, 745, 697 cm-l; lH NMR (CDCl3) ~ 7.80 (d, J =
8.7 Hz, 2H), 7.40-7.10 (m, 20 H), 7.10-7.00 (m, 8H), 6.98 (s, 2H), 6.97 (d, J = 8.3 Hz, 2H), 5.48 (m, lH), 5.15 (s, 2H), 5.09 (s, 2H), 4.79 (s, 4H), 4.74 ( m, lH), 4.72 (s, 2 H), 3.99 (dd, J = 12.3, 5.5 Hz, lH), 3.78 (dd, J = 11.0, 6.0 Hz, lH), 3.58 ( dd, J = 11.0, 3.3 Hz, lH), 3.51 (dd, J = 12.3, 2.1 Hz, lH) 2.78 (s, 3H); LRMS (M + H) 1051 (65), 797 (21), 661 (100), 571 (38); HRMS calcd for C62H55N2Ol2S (M + H) 1051.3476, found 1051.3433; Anal. Calcd. for C62H5~N2Ol2S-H20: C, 69.65; H, 5.28;
N 2.62; S, 3.00; found C, 69.79; H, 5.30; N, 2.59: S, 2.82.
Wo 94/20062 215 7 41~ PCT/US94102283 _ ~ Tran~-~-[~-~2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxy benzoyloxy]-3-(4-hydroxybenzamido)-1-Imethylsulfonyl) pyrrolidine (COMPOUND 501) To Compound 600 (323 mg, 0.307 mmol) and Pd(OH)2 (50 mg of a 20% by weight powder) under N2 were added ethanol (28 ml) then trifluoroacetic acid (28 ~1). The flask was evacuated and filled with H2 three times, then stirred under H2 27 h. The mixture was filtered through Celite, washed with methanol (40 ml) and the filtrate evaporated to a yellow glass. A DMF
solution of the reaction product was purified by reverse phase HPLC (21 x 250 mm C18 column) to provide Compound 501 (137 mg, 74%) as a yellow powder after lyophilization: mp 167-170, 179-187C (dec); IR (KBr) 3394, 1708, 1607, 1235, 762 cm ; H
NMR (CD30D) ~ 7.53 (d, J = 8.7 Hz, 2H), 7.29 (d, J = 7.6 Hz, lH), 7.07 (dd, J = 7.9, 8.1 Hz, 1 H), 6.82 (d, J = 8.2 Hz, lH),
anti-4-~3,5-Dihydroxy-~-~2-phenylcarbonyl-benzoyloxy-3-(~-hydroxybenzamido)azepine To a solution of tran-4-(3,5-dibenzyloxy-4-phenyl carbonyl-benzoyloxy-3-(4-benzyloxybenzamido)-N-benzylazepine (40 mg, 0.042 mmol) was added Pd(OH)2 (Pearlman's catalyst) 20 mg, 50% on weight basis) and introduced H2 gas at atmosphere pressure.
Trans-l-(~-hydroxybenzamido)-2-~4-benzyl-3,5-dihydroxybenzoyloxy) cycloheptane The catalyst Pd(OH)2 on carbon (20%, moist, 20 mg) was added to a solution of trans-1-(4-benzoyloxybenzamido)-2-)4-W094l2~62 PCT~S94/02283 21S7~1Z - 164 -benzoyl-3,5-dibenzyloxybenzoyloxy)cycloheptane (215 mg, 0.28 mmol) in methanol (8.4 ml). The mixture was stirred vigorously at room temperature under 1 atm H2 contained in a balloon for sixteen hours. The solid catalyst was removed by flash chromatography (sioz~ 2:2:1/diethyl ether:hexane:methylene chloride) to give a white powder (112 mg, 84%): mp 224-226-C;
lH NMR (CD30D) ~ 7.52-7.56 (m, 2H), 7.02-722 (m, 5H), 6.94 (s, 2H), 6.71-6.75 (m, 2H), 5.10 (tm, J = 9.1 Hz, lH), 4.35 (tm, J-9.3 Hz, lH`), 3.93 (s, 2H), 1.56-2.02 (m, lOH) IR (KBr) cm 3389, 1687, 1626. Anal. calcd. for C28H29O6N: C, 70.72: H, 6.15:
N, 2.95. Found C, 70.39; H, 6.37; N, 2.67.
Trans~ -hydroxybenzamido)-2-[~-(2-hydroxybenzoyl)-3,5-dihy~roxybenzoyloxy] cycloheptane The catalyst Pd(OH)2 on carbon (20%, moist, 9 mg) was added to a solution of trans-1-(4-benzyloxybenzamido)-2-t4-(2-benzyloxybenzoyl)-3,5-dibenzyloxybenzoyloxy~ cycloheptane (112 mg, 0.13 mmol) in methanol (3.9 ml) and ethyl acetate (1.3 ml).
The mixture was stirred vigorously at room temperature under 1 atm H2 contained in a balloon for seventeen hours. The solid catalyst was removed by filtration through Florisil. The filtrate was evaporated and purified by flash chromatography (SiO2 2:2:1/ethyl acetate:hexane:methylene chloride) to give a pale yellow powder (40 mg, 61~): mp 234-236-C; lH NMR (CD30D) ~ 7.56-7.59 (m, 2H), 7.47 (t, J-7.1 Hz, lH), 7.23 (d, J=8.0 Hz, lH), 7.00 (s, 2H), 6.96 (d, J=8.2 Hz, lH), 6.74-6.78 (m, 2H), 5.15 (tm, J=9.3 Hz, lH), 4.40 (tm, J=9.3 Hz, lH), 1.58-2.05 (m, lOH) IR (KBr) cm 3392, 1700, 1678, 1626. Anal. calcd. for C28H29O6N: C, 66.53; H, 5.38 N, 2.77. Found: C, 66.37; H, 5.56; N, 2.47.
wos4/20062 _ 165215 7 4 12 PCT~S94/02283 (+)-Tr~ns-4-[~-(2-Ca.LGa~ 6-hy~roxybenzoyl)-3,5- ~ihy~roxy-benzoyloxy]-3-~-hydroxybenzami~o)-1-~methylsulfonyl) pyrrolidine ~COMPO~ND 501) HO N31 H2' 10h Pd/C HO~ ~N~
~j CH30 H , ~ ~OBn N 2. THF, 2 N KOH N
- S 02CH3ClOC~OBn S 02CH3 C 02Bn NE~, l o OBn DMAP, C3~COCI
OBn HO~ N~3~ EP~dOOHHT2FA BnO~O NJ~3 h OH 0~ h OBn H SO2CH3 BnO2C~b 8BBn N
~+)-Tr~ns-~-~ydroxy-3-~-benzylGxyLr~z~mido)-l-~methylsulfonyl) -pyrrol~dine To 10% palladium on carbon (41 mg) wetted with methanol (1.0 ml) was added the azide (412 mg, 2.00 mmol) in methanol (9.0 ml). The flask was evacuated and filled with H2 twice then allowed to stir under H2 (1 atm) for 4 h. The mixture was filtered through Celite and washed through with methanol (100 ml). The methanol was evaporated, providing an off-white solid which was used without characterization.
To a O C solution of the above amino alcohol in THF
(6.0 ml) was added 2 N KOH (1.0 ml). The ice bath was removed, and the acid chloride (approx. 0.75 eq) added portionwise over 2.5 h, until the starting material was gone as evidenced by thin layer chromatography. The mixture was diluted with CH2Cl2 (30 ml) and poured into H2O (60 ml). Methanol (20 ml) was W094/2~62 PCT~S94/02283 21S7~2 - 166 -added to disperse the emulsion, the layers were separa~ed, and the aqueous layer extracted with CH2C12 (4 x 50 ml). The organic lyaers were combined, dried (MgSO4) filtered and evaporated to provide the title compound as a white solid (593 mg, 76% over two steps).lH NMR (CD30D) ~ 7.60 (d, J = 8.7 Hz, 2H), 7.25-7.10 (m, 5H), 6.85 (d, J = 9.0 Hz, 2H), 4.94 (s, 2H), 4.18-4.12 (m, 2H), 3.58-3.42 (m, 4H), 2.71 (s, 3H).
(~)-Tr~ns-~-[~-~6-benzyloxy-2-(benzyloxycarbonyl)benzoyl]-3,5-~ibenzyloxybenzoyloxy)-3-(~-~enzyloxybenzami~o)-1-~methyl sulfonyl)-pyrrolidi~e ~COMPOUND 600) To a 0-C solution of the product of the previous step (202 mg, 0.518 mmol), diisopropylethylamine (98 ~1, 1.2 eq., 0.565 mmol) and 4-dimethylaminopyridine (58 mg, 1.0 eq., 0.471 mmol) in CH2Cl2 (8.0 ml) under N2 was added a solution of the acid chloride (0.471 mmol) in CH2C12 (4.0 ml). The reaction was allowed to warm to room temperature with stirring over 16 h.
The cloudy reaction mixture was diluted with CH2Cl2 (50 ml) and washed with satd. NaHCO3 (30 ml) then brine (30 ml). The aqueous layers were extracted with CH2C12 (50 ml) each. The organics were combined, dried (MgSO4), filtered and evaporated to a light yellow oil. Flash column chromatography (1:1 hexane:ethyl acetate) provided the title product as an off white foam (371 mg, 75%): mp 73-79-C; IR (KBr) 3367, 1723, 1660, 1230, 1112, 745, 697 cm-l; lH NMR (CDCl3) ~ 7.80 (d, J =
8.7 Hz, 2H), 7.40-7.10 (m, 20 H), 7.10-7.00 (m, 8H), 6.98 (s, 2H), 6.97 (d, J = 8.3 Hz, 2H), 5.48 (m, lH), 5.15 (s, 2H), 5.09 (s, 2H), 4.79 (s, 4H), 4.74 ( m, lH), 4.72 (s, 2 H), 3.99 (dd, J = 12.3, 5.5 Hz, lH), 3.78 (dd, J = 11.0, 6.0 Hz, lH), 3.58 ( dd, J = 11.0, 3.3 Hz, lH), 3.51 (dd, J = 12.3, 2.1 Hz, lH) 2.78 (s, 3H); LRMS (M + H) 1051 (65), 797 (21), 661 (100), 571 (38); HRMS calcd for C62H55N2Ol2S (M + H) 1051.3476, found 1051.3433; Anal. Calcd. for C62H5~N2Ol2S-H20: C, 69.65; H, 5.28;
N 2.62; S, 3.00; found C, 69.79; H, 5.30; N, 2.59: S, 2.82.
Wo 94/20062 215 7 41~ PCT/US94102283 _ ~ Tran~-~-[~-~2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxy benzoyloxy]-3-(4-hydroxybenzamido)-1-Imethylsulfonyl) pyrrolidine (COMPOUND 501) To Compound 600 (323 mg, 0.307 mmol) and Pd(OH)2 (50 mg of a 20% by weight powder) under N2 were added ethanol (28 ml) then trifluoroacetic acid (28 ~1). The flask was evacuated and filled with H2 three times, then stirred under H2 27 h. The mixture was filtered through Celite, washed with methanol (40 ml) and the filtrate evaporated to a yellow glass. A DMF
solution of the reaction product was purified by reverse phase HPLC (21 x 250 mm C18 column) to provide Compound 501 (137 mg, 74%) as a yellow powder after lyophilization: mp 167-170, 179-187C (dec); IR (KBr) 3394, 1708, 1607, 1235, 762 cm ; H
NMR (CD30D) ~ 7.53 (d, J = 8.7 Hz, 2H), 7.29 (d, J = 7.6 Hz, lH), 7.07 (dd, J = 7.9, 8.1 Hz, 1 H), 6.82 (d, J = 8.2 Hz, lH),
6.73 (s, 2H), 6.62 (d, J = 8.7 Hz, 2H), 5.29 (dt, J = 5.5, 3.1 Hz, lH), 4.48 (dt, J = 6.2, 4.2 Hz, lH), 3.73 (dd, J = 12.0, 5.5 Hz, lH), 3.67 (dd, J = 10.7, 6.9 Hz, lH), 3.35 (dd, J =
12.0, 2.8 Hz, lH), 3.28 (dd, J = 10.6, 4.4 Hz, lH), 2.75 (s, 3H); LRMS (M + H) 601 (100), 301 (54), 283 (56); HRMS calcd for C27H25N2O12S (M + H) 601.1128, found 601.1831; Anal. Calcd.
for C2,H24N2O12S-2.5 H20: C, 50.23; H, 4.53; N, 4.34; S, 4.97;
found C, 50.22; H, 4.36; N, 4.37; S, 4.77.
- 168 - PCT~S94/02283 21~, -Tr~ns-~-t~-~2-Hy~roxycarbonyl-C-hy~roxybenzoyl)-3~5-~ihy~roxybenzoyloxy]-3-(~-hy~roxybenz~mi~o)-1-tphenylsulfonyl) pyrrolidine ~CONPOUND 502) HO~N~ Na~CO~ ~ ~lOBn H TFA S 02Ph f ~2Bn CH2C12 ~COCI
OBn ~0 ~N~ EtdOOH,H)2HF BnO~o N~
OH 0~ ~j OBn H2cb~ooH SO2Ph 2 ~ OBn S02Ph tl)-Tr~ns-3-(~-benzylG~ z~mi~o)-~-hy~roxy-l-(phenylsulfonyl) -pyrrolidine To a slurry of starting hydroxyamide (150 mg, 0.352 mmol) in H2O (8.8 ml) and CH2C12 (8.8 ml) were added anhydrous Na2CO3 (112 mg, 3.0 eq, 1.06 mmol) then benzenesulfonyl chloride (58 ~1, 0.458 mmol, 1.3 eq), and the mixture stirred at room temperature 15 h. The solution was then diluted with CH2C12 (20 ml) and poured into H20 (20 ml) and methanol (4 ml). The layers were separated and the aqueous layer extracted with CH2Cl2 (3 x 30 ml). The organics were combined, dried (MgSO~), filtered and evaporated to a white powder (159 mg, quant yield): H NMR (CD30D) ~ 7.62 (d, J = 7.7 Hz, 2H), 7.43 (d, J
= 8.9 Hz, 2H), 7.35-7.30 (m, 3H), 7.25-7.10 (m, 5H), 6.80 (d, J = 8.8 Hz, 2H), 4.95 (s, 2H), 4.06-4.00 (m, lH), 3.95-3.90 (m, 215~412 W094/20062 PCT~S94/02283 lH), 3.50-3.35 (m, 2H), 3.15 (dd, J = 10.6, 3.9 Hz, lH), 2.99 (dd, J = 10.8, 3.2 Hz, lH).
(+)-Tr~ns-4-[~-(6-benzyloxy-2-(benzyloxyc~rbonyl)benzoyl]-3,5-dibenzyloxybenzoyloxy)-3-(~-benzyloxybenz~mi~o)-1- (phenyl sulfonyl)-pyrrolidine (COMPOUND 601) To a solution of the prior product (159 mg, 0.352 mmol) in CH2Cl2 (6.0 ml) were added 4-dimethylaminopyridine (43 mg, 0.352~mol, 1.0 eq), diisopropylethylamine (74 ~1, 0.42 mmol, 1.2 eq) then a solution of acid chloride (0.383 mmol, 1.1 eq) in CH2Cl2 (3.0 ml). The mixture was stirred at room temperature under N2 14 h. The reaction mixture was then diluted with CH2C12 (30 ml), and washed with 10% NaHCO3 (50 ml) then brine (50 ml). The aqueous layers were combined and extracted with CH2C12 (2 x 50 ml). The organics were combined, dried (MgSO~), filtered and evaporated. Flash column chromatoghraphy of the residue (2:1 hexane:ethyl acetate) on silica gel provided the title compound (183 mg, 47%): 1H NMR
(CDCl3) ~ 7.77 (d, J = 6.7 Hz, 2H), 7.70 (d, J = 8.8 Hz, 2H),
12.0, 2.8 Hz, lH), 3.28 (dd, J = 10.6, 4.4 Hz, lH), 2.75 (s, 3H); LRMS (M + H) 601 (100), 301 (54), 283 (56); HRMS calcd for C27H25N2O12S (M + H) 601.1128, found 601.1831; Anal. Calcd.
for C2,H24N2O12S-2.5 H20: C, 50.23; H, 4.53; N, 4.34; S, 4.97;
found C, 50.22; H, 4.36; N, 4.37; S, 4.77.
- 168 - PCT~S94/02283 21~, -Tr~ns-~-t~-~2-Hy~roxycarbonyl-C-hy~roxybenzoyl)-3~5-~ihy~roxybenzoyloxy]-3-(~-hy~roxybenz~mi~o)-1-tphenylsulfonyl) pyrrolidine ~CONPOUND 502) HO~N~ Na~CO~ ~ ~lOBn H TFA S 02Ph f ~2Bn CH2C12 ~COCI
OBn ~0 ~N~ EtdOOH,H)2HF BnO~o N~
OH 0~ ~j OBn H2cb~ooH SO2Ph 2 ~ OBn S02Ph tl)-Tr~ns-3-(~-benzylG~ z~mi~o)-~-hy~roxy-l-(phenylsulfonyl) -pyrrolidine To a slurry of starting hydroxyamide (150 mg, 0.352 mmol) in H2O (8.8 ml) and CH2C12 (8.8 ml) were added anhydrous Na2CO3 (112 mg, 3.0 eq, 1.06 mmol) then benzenesulfonyl chloride (58 ~1, 0.458 mmol, 1.3 eq), and the mixture stirred at room temperature 15 h. The solution was then diluted with CH2C12 (20 ml) and poured into H20 (20 ml) and methanol (4 ml). The layers were separated and the aqueous layer extracted with CH2Cl2 (3 x 30 ml). The organics were combined, dried (MgSO~), filtered and evaporated to a white powder (159 mg, quant yield): H NMR (CD30D) ~ 7.62 (d, J = 7.7 Hz, 2H), 7.43 (d, J
= 8.9 Hz, 2H), 7.35-7.30 (m, 3H), 7.25-7.10 (m, 5H), 6.80 (d, J = 8.8 Hz, 2H), 4.95 (s, 2H), 4.06-4.00 (m, lH), 3.95-3.90 (m, 215~412 W094/20062 PCT~S94/02283 lH), 3.50-3.35 (m, 2H), 3.15 (dd, J = 10.6, 3.9 Hz, lH), 2.99 (dd, J = 10.8, 3.2 Hz, lH).
(+)-Tr~ns-4-[~-(6-benzyloxy-2-(benzyloxyc~rbonyl)benzoyl]-3,5-dibenzyloxybenzoyloxy)-3-(~-benzyloxybenz~mi~o)-1- (phenyl sulfonyl)-pyrrolidine (COMPOUND 601) To a solution of the prior product (159 mg, 0.352 mmol) in CH2Cl2 (6.0 ml) were added 4-dimethylaminopyridine (43 mg, 0.352~mol, 1.0 eq), diisopropylethylamine (74 ~1, 0.42 mmol, 1.2 eq) then a solution of acid chloride (0.383 mmol, 1.1 eq) in CH2Cl2 (3.0 ml). The mixture was stirred at room temperature under N2 14 h. The reaction mixture was then diluted with CH2C12 (30 ml), and washed with 10% NaHCO3 (50 ml) then brine (50 ml). The aqueous layers were combined and extracted with CH2C12 (2 x 50 ml). The organics were combined, dried (MgSO~), filtered and evaporated. Flash column chromatoghraphy of the residue (2:1 hexane:ethyl acetate) on silica gel provided the title compound (183 mg, 47%): 1H NMR
(CDCl3) ~ 7.77 (d, J = 6.7 Hz, 2H), 7.70 (d, J = 8.8 Hz, 2H),
7.47-7.16 (m, 14H), 7.15-7.05 (m, 6H), 7.04-6.94 (m, 3H), 6.90-6.83 (m, 3H), 6.42 (d, J = 6.7 Hz, lH), 5.30 (dt, J = 5.1, 2.6 Hz, lH), 5.18 (s, 2H), 5.13 (s, 2H), 4.78 (m, 2H), 4.76 (s, 2H), 4.72 (s, 2H), 4.64-4.60 (m, lH), 3.88 (dd, J = 12.6, 5.4 Hz, lH), 3.74-3.65 (m, lH), 3.60-3.38 (m, 2H).
(+)-Trans-~-t4-(2-Hydroxyc~rbonyl-6-hydrOXybenZOYll-3,5-dih~k6~benzoyloxy]-3-(~-hydroxybenz~mido)-l-(phenylsulfonyl) pyrrolidine (COMPOUND 502) To a solution of Compound 601 (183 mg, 0.164 mmol) in THF (7.4 ml) and ethanol (7.4 ml) was added Pd(OH)2 (92 mg, of a 20% by weight powder). The flask was evacuated and filled with H2 twice, then stirred under H2 (1 atm) for 20 h. The suspension was filtered through Celite, washed through with methanol (50 ml), and evaporated to a yellow oil. Purification by HPLC (21 x 250 mm Cl8 column) provided Compound 502 (75 mg, 69%) as a fluffy yellow powder after lyophilization: mp 185-208-C; IR (KBr) 3402, 1709, 1636, 1608, 1232 cm-l; lH NMR
WOg4/2~62 PCT~S94/02283 (CD30D) ~ 7.53 (d, J = 8.2 Hz 2H), 7.49 (d, J = 8.7 Hz, 2H), 7.32 (d, J = 7.3 Hz, lH), 7.22-7.11 (m, 3H), 7.09 (dd, J = 8.1, 7.9 Hz, lH), 6.84 (d, J = 7.2 Hz, lH), 6.61 (d, J = 8.7 Hz, 2H), 6.35 (s, 2H), 4.99 (app t, J = 2.2 Hz, lH), 4.92 (dd, J =
5.6, 2.8 Hz, lH), 3.62 (dd, J = 13.0, 4.3 Hz, lH), 3.50 (dd, J
= 10.8, 6.0 Hz, lH), 3.39 (dd, J = 10.7, 2.4 Hz, lH), 3.32 (bd, J = 13.1 Hz, lH); HRMS (M + H) calcd 663.1285, found 663.1302; Anal. Calcd. for C32H26N2Ol2S-l H2O: C, 56.47: H, 4.15;
N, 4.12; S, 4.71; found: C, 56.56; H, 4.17; N, 4.09; S, 4.58.
~09412~62 2 1 5 7 4 1 ~ PCT~S94/02283 Trans-4-[~-(2-Ethoxycarbonyl-6-hydroxybenzoyl)-3,5-~ihydroxybenzoyloxy]-3-~-hy~roxybenz~mido)-~-~methylsulfonyl) pyrrolidine ~COMPOUND 503) (1)-Tr~s-4-[4-(2-Ethoxyc~ nyl-6-hy~roxybenzoyl)-3-etho~cy-S-hy~roxybenzoyloxy~-3-(4-hydroxybenz~mido)-1-~methylsulfonyl) pyrrolidine ~COMPOUND 50~) ~h ~
E102C~oo H N
H~ohN~3~ Etl, Na2C03 503 H02C~OHsO2CH3 acetone, DMF
501 X~ h ~
EtO2C~gH N
To a solution of Compound 501 (70 mg, 0.12 mmol) in acetone (5.6 ml) under N2 were added Na2CO3 (anhyd, 25 mg, 0.23 mmol, 2.0 eq) then iodoethane (47 ~1, 0.58 mmol, 5.0 eq).
After stirring at room temperature 3h, a solid began to form.
After Sh, more iodoethane (0.47 ml, S0 eq) was added. After 22h, more iodoethane (0.47 ml, 50 eq) was added, and after another 24 h DMF (1.0 ml) was added to force the precipitate to dissolve. The clear yellow solution was stirred 20 h more, evaporated to approx. 2 ml, then partitioned between H20 (30 ml) and CH2Cl2 (30 ml). The layers were separated, and the aqueous layer extracted with CH2Cl2 (5 x 20 ml). The organics were combined, dried (MgSO4), filtered and evaporated. The yellow residue was purified by reverse phase HPLC (21 x 250 mm Cle column) to provide Compound 503 (27 mg, 37%) as a yellow 2157~12 - 172 -power after lyophilization, as well as Compound 504 (11 mg, 14%) as a light yellow powder after lyophilization.
Data for Compound 503 are mp 146-162-C; IR (KBr) 3404, 3364, 2361, 1714, 1637, 1576, 1300, 1231 cm; H NMR
(CD30D) ~ 8.33 (d, J = 6.6 Hz, lH), 7.53 (d, J = 8.7 Hz, 2H), 7.27 (d, J = 7.7 Hz, lH), 7.08 (dd, J = 8.1, 7.8 Hz, lH), 6.83 (d, J = 8.2 Hz, lH), 6.74 (s, 2H), 6.62 (d, J = 8.7 Hz, 2H), 5.29 (dd, J = 5.9, 3.2 Hz, lH), 4.53-4.47 (m, lH), 3.93 (q, J
= 7.1 Hz, 2H), 3.73 (dd, J = 12.1, 5.6 Hz, lH), 3.68 (dd, J =
10.3, 6.5 Hz, lH), 3.35 (dd, J = 12.0, 2.9 Hz, lH), 3.28 (dd, J = 10.7, 4.5 Hz, lH), 2.76 (s, 3H), 0.92 (t, J = 7.1 Hz, 3H);
LRMS (M + H) 629 (100), 555 (10), 203 (41); HRMS (M + H) calcd 629.1441, found 629.1476; Anal. Calcd. for C29H28N2Ol2S-1.25 H2O: C, 53.49; H, 4.72; N, 4.30; S, 4.92; found:
C, 53.66; H, 4.61; N, 4.29; S, 4.87.
Data for Compound 504 are mp 129-134, 140-148 C (dec);
IR (KBr) 3404, 2362, 1715, 1633, 1573, 1300, 1229 cm 1; lH NMR
(CD30D) ô 7.53 (d, J = 8.7 Hz, 2H), 7.29 (d, J = 7.7 Hz, lH), 7.12 (dd, J = 8.1, 7.9 Hz, lH), 7.00 (d, J = 1.5 Hz, lH), 6.84 (d, J = 8.8 Hz, lH), 6.76 (d, J = 1.5 Hz, lH), 6.63 (d, J = 8.8 Hz, 2H), 5.30 (dd, J = 5.4, 3.1 Hz, lH), 4.57-4.50 (m, lH), 3.92 (q, J = 7.1 Hz, 2H), 3.76-3.68 (m, 2H), 3.56 (q, J = 6.9 Hz, 2H), 3.39 (dd, J = 11.9, 3.1 Hz, lH), 3.28 (dd, J = 10.7, 4.6 Hz, lH), 2.78 (s, 3H), 0.91 (t, J = 7.1 Hz, 3H), 0.50 (t, J = 6.9 Hz, 3H); HRMS (M + H) calcd 657.1754, found 657.1619;
Anal. Calcd. for C31H32N2Ol2S-H2O: C, 55.19; H, 5.08; N, 4.15; S, 4.75; found: C, 55.18; H, 4.92; N, 4.10; S, 4.66.
`W094/20062 21~ 7 ~1~ PCT~S94/0~83 ~yn~ 3,5-Dihydroxy-~-~2-hyd o~yLa~zoyl)benzoyloxy)hex~hydro-3-~-hydroxybenzoyl~mino)azepine ~COMPOUND 505) 3-Acetyl~minohexahydro-1-phenylmethylazepin-2,~-dione A solution of hexahydro-l-phenylazepin-2,3,4-trione-3-oxime (1.23 g, 5 mmol) in 4:1 acetic acid/acetic anhydride (20 ml) was treated with Raney nickel (Aldrich, one-half tsp) in a Parr bottle and subjected to hydrogenation over 18 h at 40-45 psi and room temperature. The mixture was carefully evacuated of hydrogen and filtered through Celite. The filter pad was then washed with methanol (with care taken not to let the filter pad become dry). The filtrate was concentrated in vacuo and the residue diluted with toluene and further concentrated to remove most of the acetic acid. The residue was chilled on an ice bath and treated with saturated sodium bicarbonate carefully to avoid excessive bubbling. The cloudy aqueous solution was extracted with methylene chloride (3x50 ml, and the combined organic solution dried (Na2S04) and concentrated in vacuo. The residue was flash chromatographed on silica gel (eluted with l9:1 methylene chloride/methanol) to afford 3-acetylaminoheYA~ydro-l-phenylmethylazepin-2,4-dione (1.11 g, 81%) as a white solid.
syn-3-Aminohexahydro-~-hydroxy-l-phenylmethylazepin-2-one A solution of 3-acetylaminohexahydro-1-phenylmethylazepin-2,4-dione (0.82 g, 3.0 mmol) in absolute ethanol (15 ml) was treated with sodium borohydride (0.23 g, 6 mmol) and stirred for 30 min, then treated with water (5 ml) and concentrated in vacuo. The aqueous residue was extracted with methylene chloride (3x25 ml) and the combined organic extracts were dried (Na2SO~), concentrated in vacuo, and taken up in 2:1 ethanol/water (7.5 ml). Concentrated hydrochloric acid (2.5 ml) was added, and the mixture was refluxed for 2h and partially concentrated, then diluted with water (25 ml).
The aqueous acidic mixture was extracted with ether (25 ml), and the aqueous solution basified with 30% sodium hydroxide and extracted with methylene chloride (3x40 ml). The combined W094/2~62 PCT~S94/02283 ~157412 - 174 -methylene chloride extracts were washed with water (25ml), dried (Na2S0~), and concentrated in vacuo to a yellow solid, which was recrystallized from ethyl acetate to afford syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.42 g, 60%) as a white solid.
syn-3-Aminohexahydro-4-hydroxy-1-phenylmethylazepine A cooled (5-C) solution of lithium aluminum hydride/tetrahydrofuran (Aldrich, 1.0 N, 5.1 ml) under nitrogen was treated with syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.40 g, 1.7 mmol) in portions so that the pot temperature did not exceed 15-C. The mixture was refluxed for 6.5h, cooled on an ice bath, and carefully treated with water (0.21 ml), 15% sodium hydroxide (0.21 ml), and water (0.63 ml). The suspension was allowed to stir for 5 days, during which time the product partially decomposed (optimal time is 2 - 5 hours). The suspension was filtered, and the filtrate was concentrated in vacuo and chromotographed on silica gel (eluted with 90:8:2 methylene chloride/methanol/triethylamine). The appropriate fractions were concentrated in vacuo to afford syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.22 g, 58%) as a colorless oil.
syn-Hexahydro-~-hydroxy-3-~-phenylmethoxy)benzoylamino-1-phenylmethylazepine A solution of 4-benzyloxybenzoic acid (0.183 g, 0.8 mmol) in anhydrous tetrahydrofuran (2 ml) and N,N-dimethylformamide (0-5 ml) was treated with N~Nl-carbonyldiimidazole (0.15 g, 0.9 mmol) and stirred at room temperature for 1.5h. The solution was treated with syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.20 g, 0.9 mmol) in anhydrous tetrahydrofuran (1 ml), and the mixture was stirred for 18 h, then concentrated in vacuo. The residue was taken up in lN sodium carbonate (20 ml), and the aqueous mixture was extracted with toluene (2x25 ml) containing a little 2-propanol. The combined organic extracts were dried (Na2S0~) and the concentrated residue was flash chromatographed on silica gel (eluted with 3:1 ethyl acetate/hexane) to afford syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.17 g, 50%) as a viscous oil.
3,5-B~ 8 (phenylmethoxy)-~.-(2-phenylmethoxybenzoyl)benzoic acid ester with syn-}lexahy~ro-~-hy~roxy-3-(~-phenylmethoxy)benzoyl ~no-l-phenylmethylazepine A solution of 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)benzoic acid (0.245 g, 0.45 mmol) in anhydrous methylene chloride (1.5 ml) was treated with N,N-dimethylformamide (2 drops), then with 2.0 N oxalyl chloride/methylene chloride (Aldrich, 0.30 ml, 0.60 mmol), and stirred for one hour under nitrogen. The solution was concentrated in vacuo and placed under high vacuum for one hour. syn-Hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.170 g, 0.40 mmol) was dissolved in anhydrous methylene chloride (3 ml), treated with 4-dimethylaminopyridine (0.001 g) and triethylamine (0.12 ml, 1.2 mmol), and cooled on an ice bath under nitrogen. The acid chloride was removed from high vacuum and dissolved in anhydrous methylene chloride (2 ml), then added to the cooled solution, and the mixture was allowed to warm to room temperature, stirred for one hour, and partially concentrated in vacuo. The residual solution was deposited on a silica gel column and eluted (first with 2:1 hexane/methylene chloride, then with 1:1 hexane/methylene chloride) to afford (after concentration of the appropriate fractions) 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)benzoic acid ester with syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.29 g, 77%) as a white foam.
Qyn-(~.-(3,5-Dih~0~c",~ 4-(2-hy~.c,J,~e..zoyl)benzoyloxy))hexahydro -3-(~-hydroxybenzoyl~ino)~zepine (COMPO~ND 505) - A cloudy suspension of 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)benzoic acid ester with syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.29 g, 0.30 mmol) in reagent ethanol (25 ml) was treated with 2 La ~ ~12 -176 - ~-20% Pd(OH)2/C (Aldrich, Pearlman's catalyst, 0.20 g) in a Parr bottle, then subjected to hydrogenation for 24h at 50 - 52 psi.
The hydrogen was evacuated and the solution was carefully filtered through Celite under nitrogen, and the filter pad was washed with methanol (not to dryness). The filtrate was concentrated in vacuo to crude material, which was flash chromatographed on a short column of silica gel (eluted with 1:1 CHCl3/EtOH) to afford 0.13 g of product as a pale yellow foam. This was triturated from ether/acetonitrile to afford syn- ( 4-(3,5-dihydroxy-4-(2-hydroxybenzoyl) benzoyloxy)) hexahydro-3-(4-hydroxybenzoylamino)azepine (0.95 g, 62%) as a pale yellow powder (dihydrate); mp 177-179-C. R (1 1 CHCl3/EtOH on silica) 0.45; IR (KBr): 1623 cm 1; ~H NMR (d6-DMSO)
(+)-Trans-~-t4-(2-Hydroxyc~rbonyl-6-hydrOXybenZOYll-3,5-dih~k6~benzoyloxy]-3-(~-hydroxybenz~mido)-l-(phenylsulfonyl) pyrrolidine (COMPOUND 502) To a solution of Compound 601 (183 mg, 0.164 mmol) in THF (7.4 ml) and ethanol (7.4 ml) was added Pd(OH)2 (92 mg, of a 20% by weight powder). The flask was evacuated and filled with H2 twice, then stirred under H2 (1 atm) for 20 h. The suspension was filtered through Celite, washed through with methanol (50 ml), and evaporated to a yellow oil. Purification by HPLC (21 x 250 mm Cl8 column) provided Compound 502 (75 mg, 69%) as a fluffy yellow powder after lyophilization: mp 185-208-C; IR (KBr) 3402, 1709, 1636, 1608, 1232 cm-l; lH NMR
WOg4/2~62 PCT~S94/02283 (CD30D) ~ 7.53 (d, J = 8.2 Hz 2H), 7.49 (d, J = 8.7 Hz, 2H), 7.32 (d, J = 7.3 Hz, lH), 7.22-7.11 (m, 3H), 7.09 (dd, J = 8.1, 7.9 Hz, lH), 6.84 (d, J = 7.2 Hz, lH), 6.61 (d, J = 8.7 Hz, 2H), 6.35 (s, 2H), 4.99 (app t, J = 2.2 Hz, lH), 4.92 (dd, J =
5.6, 2.8 Hz, lH), 3.62 (dd, J = 13.0, 4.3 Hz, lH), 3.50 (dd, J
= 10.8, 6.0 Hz, lH), 3.39 (dd, J = 10.7, 2.4 Hz, lH), 3.32 (bd, J = 13.1 Hz, lH); HRMS (M + H) calcd 663.1285, found 663.1302; Anal. Calcd. for C32H26N2Ol2S-l H2O: C, 56.47: H, 4.15;
N, 4.12; S, 4.71; found: C, 56.56; H, 4.17; N, 4.09; S, 4.58.
~09412~62 2 1 5 7 4 1 ~ PCT~S94/02283 Trans-4-[~-(2-Ethoxycarbonyl-6-hydroxybenzoyl)-3,5-~ihydroxybenzoyloxy]-3-~-hy~roxybenz~mido)-~-~methylsulfonyl) pyrrolidine ~COMPOUND 503) (1)-Tr~s-4-[4-(2-Ethoxyc~ nyl-6-hy~roxybenzoyl)-3-etho~cy-S-hy~roxybenzoyloxy~-3-(4-hydroxybenz~mido)-1-~methylsulfonyl) pyrrolidine ~COMPOUND 50~) ~h ~
E102C~oo H N
H~ohN~3~ Etl, Na2C03 503 H02C~OHsO2CH3 acetone, DMF
501 X~ h ~
EtO2C~gH N
To a solution of Compound 501 (70 mg, 0.12 mmol) in acetone (5.6 ml) under N2 were added Na2CO3 (anhyd, 25 mg, 0.23 mmol, 2.0 eq) then iodoethane (47 ~1, 0.58 mmol, 5.0 eq).
After stirring at room temperature 3h, a solid began to form.
After Sh, more iodoethane (0.47 ml, S0 eq) was added. After 22h, more iodoethane (0.47 ml, 50 eq) was added, and after another 24 h DMF (1.0 ml) was added to force the precipitate to dissolve. The clear yellow solution was stirred 20 h more, evaporated to approx. 2 ml, then partitioned between H20 (30 ml) and CH2Cl2 (30 ml). The layers were separated, and the aqueous layer extracted with CH2Cl2 (5 x 20 ml). The organics were combined, dried (MgSO4), filtered and evaporated. The yellow residue was purified by reverse phase HPLC (21 x 250 mm Cle column) to provide Compound 503 (27 mg, 37%) as a yellow 2157~12 - 172 -power after lyophilization, as well as Compound 504 (11 mg, 14%) as a light yellow powder after lyophilization.
Data for Compound 503 are mp 146-162-C; IR (KBr) 3404, 3364, 2361, 1714, 1637, 1576, 1300, 1231 cm; H NMR
(CD30D) ~ 8.33 (d, J = 6.6 Hz, lH), 7.53 (d, J = 8.7 Hz, 2H), 7.27 (d, J = 7.7 Hz, lH), 7.08 (dd, J = 8.1, 7.8 Hz, lH), 6.83 (d, J = 8.2 Hz, lH), 6.74 (s, 2H), 6.62 (d, J = 8.7 Hz, 2H), 5.29 (dd, J = 5.9, 3.2 Hz, lH), 4.53-4.47 (m, lH), 3.93 (q, J
= 7.1 Hz, 2H), 3.73 (dd, J = 12.1, 5.6 Hz, lH), 3.68 (dd, J =
10.3, 6.5 Hz, lH), 3.35 (dd, J = 12.0, 2.9 Hz, lH), 3.28 (dd, J = 10.7, 4.5 Hz, lH), 2.76 (s, 3H), 0.92 (t, J = 7.1 Hz, 3H);
LRMS (M + H) 629 (100), 555 (10), 203 (41); HRMS (M + H) calcd 629.1441, found 629.1476; Anal. Calcd. for C29H28N2Ol2S-1.25 H2O: C, 53.49; H, 4.72; N, 4.30; S, 4.92; found:
C, 53.66; H, 4.61; N, 4.29; S, 4.87.
Data for Compound 504 are mp 129-134, 140-148 C (dec);
IR (KBr) 3404, 2362, 1715, 1633, 1573, 1300, 1229 cm 1; lH NMR
(CD30D) ô 7.53 (d, J = 8.7 Hz, 2H), 7.29 (d, J = 7.7 Hz, lH), 7.12 (dd, J = 8.1, 7.9 Hz, lH), 7.00 (d, J = 1.5 Hz, lH), 6.84 (d, J = 8.8 Hz, lH), 6.76 (d, J = 1.5 Hz, lH), 6.63 (d, J = 8.8 Hz, 2H), 5.30 (dd, J = 5.4, 3.1 Hz, lH), 4.57-4.50 (m, lH), 3.92 (q, J = 7.1 Hz, 2H), 3.76-3.68 (m, 2H), 3.56 (q, J = 6.9 Hz, 2H), 3.39 (dd, J = 11.9, 3.1 Hz, lH), 3.28 (dd, J = 10.7, 4.6 Hz, lH), 2.78 (s, 3H), 0.91 (t, J = 7.1 Hz, 3H), 0.50 (t, J = 6.9 Hz, 3H); HRMS (M + H) calcd 657.1754, found 657.1619;
Anal. Calcd. for C31H32N2Ol2S-H2O: C, 55.19; H, 5.08; N, 4.15; S, 4.75; found: C, 55.18; H, 4.92; N, 4.10; S, 4.66.
`W094/20062 21~ 7 ~1~ PCT~S94/0~83 ~yn~ 3,5-Dihydroxy-~-~2-hyd o~yLa~zoyl)benzoyloxy)hex~hydro-3-~-hydroxybenzoyl~mino)azepine ~COMPOUND 505) 3-Acetyl~minohexahydro-1-phenylmethylazepin-2,~-dione A solution of hexahydro-l-phenylazepin-2,3,4-trione-3-oxime (1.23 g, 5 mmol) in 4:1 acetic acid/acetic anhydride (20 ml) was treated with Raney nickel (Aldrich, one-half tsp) in a Parr bottle and subjected to hydrogenation over 18 h at 40-45 psi and room temperature. The mixture was carefully evacuated of hydrogen and filtered through Celite. The filter pad was then washed with methanol (with care taken not to let the filter pad become dry). The filtrate was concentrated in vacuo and the residue diluted with toluene and further concentrated to remove most of the acetic acid. The residue was chilled on an ice bath and treated with saturated sodium bicarbonate carefully to avoid excessive bubbling. The cloudy aqueous solution was extracted with methylene chloride (3x50 ml, and the combined organic solution dried (Na2S04) and concentrated in vacuo. The residue was flash chromatographed on silica gel (eluted with l9:1 methylene chloride/methanol) to afford 3-acetylaminoheYA~ydro-l-phenylmethylazepin-2,4-dione (1.11 g, 81%) as a white solid.
syn-3-Aminohexahydro-~-hydroxy-l-phenylmethylazepin-2-one A solution of 3-acetylaminohexahydro-1-phenylmethylazepin-2,4-dione (0.82 g, 3.0 mmol) in absolute ethanol (15 ml) was treated with sodium borohydride (0.23 g, 6 mmol) and stirred for 30 min, then treated with water (5 ml) and concentrated in vacuo. The aqueous residue was extracted with methylene chloride (3x25 ml) and the combined organic extracts were dried (Na2SO~), concentrated in vacuo, and taken up in 2:1 ethanol/water (7.5 ml). Concentrated hydrochloric acid (2.5 ml) was added, and the mixture was refluxed for 2h and partially concentrated, then diluted with water (25 ml).
The aqueous acidic mixture was extracted with ether (25 ml), and the aqueous solution basified with 30% sodium hydroxide and extracted with methylene chloride (3x40 ml). The combined W094/2~62 PCT~S94/02283 ~157412 - 174 -methylene chloride extracts were washed with water (25ml), dried (Na2S0~), and concentrated in vacuo to a yellow solid, which was recrystallized from ethyl acetate to afford syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.42 g, 60%) as a white solid.
syn-3-Aminohexahydro-4-hydroxy-1-phenylmethylazepine A cooled (5-C) solution of lithium aluminum hydride/tetrahydrofuran (Aldrich, 1.0 N, 5.1 ml) under nitrogen was treated with syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.40 g, 1.7 mmol) in portions so that the pot temperature did not exceed 15-C. The mixture was refluxed for 6.5h, cooled on an ice bath, and carefully treated with water (0.21 ml), 15% sodium hydroxide (0.21 ml), and water (0.63 ml). The suspension was allowed to stir for 5 days, during which time the product partially decomposed (optimal time is 2 - 5 hours). The suspension was filtered, and the filtrate was concentrated in vacuo and chromotographed on silica gel (eluted with 90:8:2 methylene chloride/methanol/triethylamine). The appropriate fractions were concentrated in vacuo to afford syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.22 g, 58%) as a colorless oil.
syn-Hexahydro-~-hydroxy-3-~-phenylmethoxy)benzoylamino-1-phenylmethylazepine A solution of 4-benzyloxybenzoic acid (0.183 g, 0.8 mmol) in anhydrous tetrahydrofuran (2 ml) and N,N-dimethylformamide (0-5 ml) was treated with N~Nl-carbonyldiimidazole (0.15 g, 0.9 mmol) and stirred at room temperature for 1.5h. The solution was treated with syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.20 g, 0.9 mmol) in anhydrous tetrahydrofuran (1 ml), and the mixture was stirred for 18 h, then concentrated in vacuo. The residue was taken up in lN sodium carbonate (20 ml), and the aqueous mixture was extracted with toluene (2x25 ml) containing a little 2-propanol. The combined organic extracts were dried (Na2S0~) and the concentrated residue was flash chromatographed on silica gel (eluted with 3:1 ethyl acetate/hexane) to afford syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.17 g, 50%) as a viscous oil.
3,5-B~ 8 (phenylmethoxy)-~.-(2-phenylmethoxybenzoyl)benzoic acid ester with syn-}lexahy~ro-~-hy~roxy-3-(~-phenylmethoxy)benzoyl ~no-l-phenylmethylazepine A solution of 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)benzoic acid (0.245 g, 0.45 mmol) in anhydrous methylene chloride (1.5 ml) was treated with N,N-dimethylformamide (2 drops), then with 2.0 N oxalyl chloride/methylene chloride (Aldrich, 0.30 ml, 0.60 mmol), and stirred for one hour under nitrogen. The solution was concentrated in vacuo and placed under high vacuum for one hour. syn-Hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.170 g, 0.40 mmol) was dissolved in anhydrous methylene chloride (3 ml), treated with 4-dimethylaminopyridine (0.001 g) and triethylamine (0.12 ml, 1.2 mmol), and cooled on an ice bath under nitrogen. The acid chloride was removed from high vacuum and dissolved in anhydrous methylene chloride (2 ml), then added to the cooled solution, and the mixture was allowed to warm to room temperature, stirred for one hour, and partially concentrated in vacuo. The residual solution was deposited on a silica gel column and eluted (first with 2:1 hexane/methylene chloride, then with 1:1 hexane/methylene chloride) to afford (after concentration of the appropriate fractions) 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)benzoic acid ester with syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.29 g, 77%) as a white foam.
Qyn-(~.-(3,5-Dih~0~c",~ 4-(2-hy~.c,J,~e..zoyl)benzoyloxy))hexahydro -3-(~-hydroxybenzoyl~ino)~zepine (COMPO~ND 505) - A cloudy suspension of 3,5-bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)benzoic acid ester with syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.29 g, 0.30 mmol) in reagent ethanol (25 ml) was treated with 2 La ~ ~12 -176 - ~-20% Pd(OH)2/C (Aldrich, Pearlman's catalyst, 0.20 g) in a Parr bottle, then subjected to hydrogenation for 24h at 50 - 52 psi.
The hydrogen was evacuated and the solution was carefully filtered through Celite under nitrogen, and the filter pad was washed with methanol (not to dryness). The filtrate was concentrated in vacuo to crude material, which was flash chromatographed on a short column of silica gel (eluted with 1:1 CHCl3/EtOH) to afford 0.13 g of product as a pale yellow foam. This was triturated from ether/acetonitrile to afford syn- ( 4-(3,5-dihydroxy-4-(2-hydroxybenzoyl) benzoyloxy)) hexahydro-3-(4-hydroxybenzoylamino)azepine (0.95 g, 62%) as a pale yellow powder (dihydrate); mp 177-179-C. R (1 1 CHCl3/EtOH on silica) 0.45; IR (KBr): 1623 cm 1; ~H NMR (d6-DMSO)
8.20 (d, lH, J = 8 Hz), 7.65 (d, 2H, J = 9 Hz), 7.57 (dt, lH, J = 8, 1.5 Hz), 7.29 (dd, lH, J = 8, 1.5 Hz), 7.10 (s, 2H), 7.02 (d, lH, J = 8 Hz), 6.91 (t, lH, J = 8 Hz), 6.78 (d, 2H, J
= 9 Hz), 5.39 (br d, lH, J = 7 Hz), 4.48 (m, lH), 3.00 - 3.20 (m, 4H), 2.05 - 2.20 (m, lH), 1.70 - 2.00 (m, 3H).Anal.
Calcd. for C27H26N20~-2H2O: C, 59.77; H, 5.57; N, 5.16. Found:
C, 59.83; H, 5.39; N, 5.46.
W094l20062 21 S 7 412 PCT~S94tO~83 Anti-4-t3,5-Dihy~roxy-~-(2-hy~roxyphenylc~rbonyl)]benzoyloxy-3-(~-hydroxybenz~mido)azepine ~COMPOUND 506) To a solution of anti-3-(4-benzyloxybenzamido)-4-[3,5-dibenzyloxy-4-(2-benzyloxyphenylcarbonyl)~benzoyloxy-N-benzylazepine (150 mg, 0.156 mmol) in EtOAc/EtOH (6 ml, 1:1) was added Pd(OH)2 (Pearlman's catalyst) (90 mg, 60% on weight basis), and then H2 at atmospheric pressure. After stirring vigorously for 24h at room temperature, the reaction mixture was filtered through a pad of celite. The filtrate was concentrated and purified on flash column (silica gel: 50 ml;
eluted with 20% ethanol in methylene chloride). CompoUnd 506 was obtained as yellow powder (30 mg, 38%): mp 174 - 176-C; lH
NMR (DMSO) ~ 7.65 (d, J = 8.64, 2H, ArH), 7.54 (td, lH, ArH), 7.26 (dd, J = 1.6, 7.9 Hz, lH, ArH), 6.70 (d, J = 6.48, lH, ArH), 6.98 (s, 2H, ArH), 6.87 (td, lH, ArH), 6.77 (d, J = 8.67 Hz, 2H, ArH), 5.18 (m, lH, 4CH), 4.19 (m, lH, 3 CH), 2.94 -2.88 (dd, lH, CH2N), 2.83 - 2.73 (m, 3H, CH2N), 1.91 (m, 2H, 6 CH2), 1.74 and 1.64 (m and m, 2H, 5 CH2); IR (KBr) cm1 3394, 1704, 1623, 1609, and 1504. Anal. calcd. for C2~H26N2O8-1 1/4 H2O: C, 61.30; H, 5.43; N, 5.29. Found: C, 61.33; H, 5.29;
N, 4.96.
W094l2~62 PCT~S94/0~83 21~7~12 - 178 -Tran~ -benzyloxybenz~mido)-2-t~-(2-benzyloxycarbonylbenzo yl)-3,5-dibenzyloxybenzoyloxyl)cyclohept~ne (COMPOUND 507) An oven-dried 25ml 3-neck round bottom flask, under N2~ was charged with trans-2-(4-benzyloxybenzamido) cycloheptanol (204 mg, 0.6 mmol), 4-(2-benzyloxycarbonyl benzoyl)-3,5-dibenzyloxybenzoic acid (376 mg, 0.66 mmol), 1,3-dicyclohexylcarbodiimide (136 mg, 0.66 mmol), 4-dimethyl-aminopyridine (74 mg, 0.66 mmol), and dry methylene chloride (4.5 ml). The resultant suspension was stirred at room temperature for 17 hours, diluted with methylene chloride (6ml), washed with water (5 ml x 3), and dried MgSO4. The solvent was evaporated and the residue was chromatographed (SiO2, 1:1:2/ diethyl ether:methylene chloride:hexane) to give a white powder (429 mg, 80%): mp 153-154-C; ~H NMR (CDCl3) 6.89 - 7.68 (m, 30H), 6.33 (d, J = 8.6 Hz, lH), 5.16 (s, 2H), 5.13 (tm, J = 9.4 Hz, lH), 5.04 (s, 2H), 4.95 (ABq, J = 14.2, 12.2 Hz, 4H), 4.45 (tm, J = 9.3 Hz, lH), 1.55 - 2.08 (m, 10H) IR KBr 3466, 3367, 1735, 1717, 1679 cm 1. Anal. calcd for CS7HS1O~ C, 76.58; H, 5.75; N, 1.57. Found: C, 76.61; H, 5.82; N, 1.35.
21~7~1~
WOg4/20062 - 179 - PCT~S94/02283 Tr~ns~ -(2-Carboxy-6-hy~roxybenzoyl)-3,5-dihydroxy benzoyloxy)-3-(~-hy~roxybenz~mido)~zepineTr~fluoroaceticAci~
8alt (COMPO~ND 508) Trifluoroacetic Aci~ 8alt of Balanol BnO2C~
o~
C02~n BnO~_~ OBn B~ 1~ (COCI)2 ~ OBn r¦ T ~ 2) NEt3, DMAP, r HO2C~OBnOBn ~' N~ OBn ~ H~ OBn HO~
O ~
H2 ~ o =( OH OH
Pd(OH)2 \ OH
HO ~ ~ CF3CO2H
o O HN o (~NH 508 Trans-N-Benzyl~ -(6-benzyloxy-2-(benzyloxyc~rh~nyl)benzoyl)-3,5-~ih6nzyloxybenzoyloxy)-3-(4-benzyloxybenzamido)azepinQ
(COMPOUND 602) A solution of 84 mg (0.12 mmol) of 4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoic acid in 2 ml of methylene chloride containing a trace (approximately 0.5 ~L) of dimethylformamide was cooled to 0-C. Oxalyl chloride (11.9 ~L, 0.136 mmol) was added, and the mixture was stirred under a nitrogen atmosphere for 1.5 h. An additional 11.9 ~L
of oxalyl chloride was added, and the mixture was stirred for an additional 1.5 h. The reaction mixture was evaporated, and the residue was evaporated twice from 15 ml of methylene chloride. The residue was dissolved in 2 ml of methylene chloride, and was added to a solution of 59.1 mg (0.137 mmol) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine, 19.0 ~1 (0.136 mmol) of triethylamine, and 3 mg of DMAP in 1.5 W094l2~62 PCT~S94/02283 2157~12 - 180 _ ml of methylene chloride at 0CC. The mixture was stirred at room temperature under a nitrogen atmosphere for 22 h, after which it was diluted with 30 ml of methylene chloride, washed with saturated sodium bicarbonate and brine, dried over magnesium sulfate, and evaporated to give 139 mg of the crude product. Chromatography on silica gel eluting with 6/4 hexane - ethyl acetate gave 89.4 mg (66%) of Compound 602 as a yellow oil, which was used directly in the next step.
Tr~ns-4-(4-(2-CarhoYy-6-hy~roxybenzoyl)-3,5-di~y ~x~oenzoyloxy) -3-(4-hydroxy~6n~Amido)azepine Trifluoroacetic Acid Salt (COMPO~ND 508) A solution of 65 mg (0.060 mmol) of Trans-N-benzyl-4-(4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoyloxy)-3-(4-benzyloxybenzamido)azepinein30ml of 1/2/2 methanol/ethanol/methylene chloride was treated with 17 ~1 of trifluoroacetic acid and evaporated. The residue was dissolved in 12 ml of 3/1 ethanol/methanol, 15.6 mg of moist 10% palladium hydroxide on carbon was added, and the mixture was ch~ken on a Parr apparatus under 50 psi of hydrogen for 5 h. The mixture was filtered, evaporated, and the residue was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile;
gradient: 0-100% B over 60 min, flow: 15 ml/min). The pure fraction was evaporated and then lyophilized from water to give 11.2 mg (33%) of the title compound as a yellow fluffy solid.
IR (KBr) : 1701, 1674, 1636, 1608, 1425, 1234, 1200 cm-l;
FABMS: m/z 573 (M + Na), 551 (M+H); HRMS: calcd for C28H27N2Olo 551.1665, found 551.1697.
WO 94/20062 - 181 21~ 7 ~12 PCT/US94/02283 ent-B~l~nol (coMpo~ D 509 ) E3n,N~'NH $~CF3 (N~ oCH33 ~%&F3 ~ Cl Bn NH + Bn NH
~OB CH2CI2 ~ d~
OBn OBn 1) SilicaChi~""atography 3% NEt3, 30% EtOAc `~ in hexanes 2) KOH, MeOH
~-OH ~OH
B,N ~ B~N ~
od~ o5~
OBn OBn HO2C~
~OH ~
Bn,N~NH ~HO~_ oOH
o ~ ~d 509 ~ 0~
OBn ~~~. N~ OH
HN
~8)-Mosher's a¢id chloride Note that the chirality label is changed going from acid to acid chloride. (R)- (+)--Methoxy--trifluoromethyl acetic acid (5.1 g, 21.8 mmol) was slurried in hexane (3 ml).
DMF (2 drops) was added followed by a 2M methylene chloride solution of oxalyl chloride (33 ml, 66 mmol). The solution was refluxed for 3 h, cooled to rt, concentrated and distilled by W094/20062 PCT~S94/0~83 21S7~12 - 182 -kugelrohr (T = 45C at 0.1 mm Hg) to give the product as a clear oil.
Trans-N-Benzyl-3-~-benzyloxybenz~mido)-~ R)-2-methoxy-2-trifluoromethylacetoxy)hex~methyleneimine Trans-N-Benzyl-3-(4-benzyloxybenzamido)-4-hydroxy-hexamethyleneimine (1.2 g, 2.79 mmol), DMAP (4 mg, 0.3 mmol) and triethylamine (2.25 g, 3.1 ml, 22.3 mmol) were dissolved in methylene chloride (10 ml) and treated with (S)-Mosher's acid chloride (1.8 g, 1.3 ml, 7 mmol). When TLC indicated complete reaction, the mixture was concentrated and flashed (7 x 15 cm, 3% triethylamine in 4/1 : ethyl acetate/hexanes). The products were separated into clean upper, mixed and clean lower fractions. All three fractions were each again chromatographed on a Dynamax-60 silica column (41.4 mm ID X 30 cm length) using a linear gradient from 20% to 60% B (A = hexanes, B = 10%
triethyl amine in ethyl acetate) over 60 m at 25 ml/min. The clean upper HPLC fractions from the upper and mixed runs were combined (490 mg) for hydrolysis lH-NMR (300 MHz, CDCl3) 1.6-1.8 (2H, m), 1.94 (2H, m), 2.48 (lH, m), 2.77 (lH, m), 2.9-3.0 (2H, m), 3.50 (lH, d, J = 13 Hz), 3.54 (3H, s), 3.72 (lH, d, J = 13 Hz), 4.1-4.2 (lH, m), 5.14 (2H, s), 5.28 (lH, m), 6.84-7.65 (14H, m).
The clean lower HPLC fractions from the lower run were combined (260 mg) for hydrolysis. H-NMR (300 MHz, CDCl3) 1.64-1.8 (2H, m), 1.8-1.94 (2H, m), 2.53 (lH, m), 2.77 (lH, m), 2.9-3.0 (2H, m), 3.50 (lH, d, J = 13 Hz), 3.52 (3H, s), 3.72 (lH, d, J = 13 Hz), 4.1 (lH, m), 5.13 (2H, s), 5.28 (lH, m), 6.84-7.54 (14H, m).
The upper ester fraction (490 mg, 0.76 mmol) was dissolved in methanol (5 ml) and treated with 85% potassium hydroxide (97 mg, 1.52 mmol) dissoved in methanol (5 ml) and stirred for 16 h. The mixture was treated with water (15 ml) and extracted with methylene chloride (2 x 25 ml). The organic layer was concentrated and chromatographed (2.5 x 10 cm, ethyl acetate) to give the chiral alcohol (266 mg) as an oil.
W094/2~62 2 1 5 7 4 1 2 PCT~S94tO~
The lower ester fraction (260 mg, 0.40 mmol) was dissolved in methanol (5 ml) and treated with 85% potassium hydroxide (53 mg, 0.8 mmol) dissoved in methanol (5 ml) and stirred for 48 h. The mixture was treated with water (15 ml) and extracted with methylene chloride (2 x 25 ml). The organic layer was concentrated and chromatographed (2.5 x 10 cm, ethyl acetate) to give the chiral alcohol (154 mg) as an oil.
Balanol benzophenone (255 mg, 375 ~mol) was dissolved in methylene chloride (3 ml) and treated with DMF (3 drops) followed by a 2M methylene chloride solution of oxalyl chloride (244 ~L, 62 mg, 488 ~mol). After stirring for 1 h, the mixture was concentrated and put under vacuum. The residue was dissolved in methylene chloride (5 ml) and added to chiral amidoalcohol (150 mg, 375 ~mol), DMAP (5 mg), triethylamine (157 ~L, 114 mg, 1.13 mmol) in methylene chloride (5 ml).
After stirring for 16 h, the mixture was chromatographed directly (silica gel, 2.5 x 10 cm, 2/3 : ethyl acetate /hexanes) to give the ester (150 mg)as a glass. The glass was dissolved in 1/1 ethanol/methanol (10 ml), treated with trifluoroacetic acid (100 ~L) and Pearlman's catalyst (palladium hydroxide, 15 mg) and stirred under a hydrogen atmosphere (balloon) for 16 h. The catalyst was filtered off and the mixture concentrated. The residue was chromatographed on a Dynamax-60 C18 column (21 X 250 mm) using a linear gradient from 100% A (0.1% TFA and 5~ acetonitrile in water) to 100% B (pure acetonitrile) over 60 m at 15 ml/min. The clean product, which eluted in 24 m., was concentrated to remove acetonitrile and freeze-dried to give a light yellow powder (45 mg, 22%), Compound 509, identical to Balanol by NMR, IR, CHN
and analytical HPLC. Different rotation [~]D25 = +97.8 (c =
0.319 in CH30H).
PCT~S94/02283 ~57~ 184 -~-)-Trans~ -(2-carboxy-6-hy~roxybenzoyl)-3,5-dihy~roxy benzoyloxy)-3-(~-hy~roxybenz~mi~o)azepineTrifluoroaceticAci~
8alt, (-)-Balanol (COMr~uNv 510) 8nO
) (COCI)2 ~ ~ OBn ll l 2) IPr2NEt, DMAP, I
HO2C ~OBnOBn CH ~ OBn ~ H~ OBn Ho2~3 HO~ OH
H2 ~ OH
pd(OH)2 o ~
~NS' HO~ OH
H
Trans-N-Benzyl-~(~-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl) -3,5-~ibenzyloxybenzoyloxy)-3-(~-benzyloxybenz~mido)azepine A solution of 356 mg (0.583 mmol) of 4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoic acid in 10 ml of methylene chloride contAjning a trace (approximately 1 ~L) of dimethylformamide was cooled to O-C. A 2.0 M solution of oxalyl chloride (0.35 ml, 0.70 mmol) was added, and the mixture was stirred under a nitrogen atmosphere for 2 h. An additional 0.35 ml of oxalyl chloride was added, and the mixture was stirred for an additional 1 h. The reaction mixture was evaporated, and the residue was evaporated twice from 20 ml of methylene chloride. The residue was dissolved in 5 ml of methylene chloride, and was added to a solution of 251 mg (0.583 mmol) of trans-N-benzyl-3-(4-benzyloxybenzamido) -4-hydroxyazepine, 122 ~L (0.700 mmol) of diisopropyl-~094/2~62 215 7 412 PCT~S94/02283 ethylamine, and 4.1 mg of DMAP in 9 ml of methylene chloride at 0-C. The mixture was stirred at room temperature under a nitrogen atmosphere for 16 h, after which it was diluted with 75 ml of methylene chloride, washed with saturated sodium bicarbonate and brine, dried over magnesium sulfate, and evaporated to give 690 mg of the crude product. Chromatography on silica gel eluting with 1/1 hexane - ethyl acetate gave 381 mg (60%) of the title compound as a yellow oil. IR (KBr) :
1719, 1655, 1605, 1581, 1456, 1321, 1248, 1111, 744, 697 cm-l.
Anal. Calcd for C~oH62N2Olo-l~5 H2O: C, 75.18; H, 5.86; N, 2.51.
Found: C, 75.16; H, 5.88; N, 2.74.
~-)-Trans~ 2-Carboxy-6-hydroxybenzoyl)-3,5-dihy~roxy benzoylo~y)-3-(~-hydroxybenz~mi~o)azepine tr~fluoroaceticacid ~lt (COHPOUND 510) A solution of 363 mg (0.333 mmol) of trans-N-benzyl-4-(4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-~ihe~yloxybenzoyloxy)-3-(4-benzyloxybenzamido)azepinein30ml of ethanol was treated with 31 ~L of trifluoroacetic acid, cooled to 0-C, and 53.3 mg of moist 10% palladium hydroxide on carbon was added. The mixture was stirred under an atmosphere of H2 for 22 h. The mixture was filtered, evaporated, and the residue was chromatographed on a 41 x 250 mm C18 column (solvent A; 95:5 water/acetonitrile +0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-50% B over 60 min, flow: 25 ml/min).
The pure fractions were pooled and evaporated and then lyophilized from water to give 75.7 mg (32%) of Compound 510 as a yellow fluffy solid, together with an additional 69 mg (29%) of material which was 96% pure by NMR. mp>200 C; [~]25D = -104- (c = 0.111, methanol); IR (KBr) : 1679, 1607, 1509, 1426, 1369, 1241, 1202, 763 cm . Anal. Calcd for C28H26N2O1o 3 H2O -TFA: C, 50.14; H, 4.63, N, 3.90. Found: C, 50.11; H, 4.40; N, 4.01.
W094/2~62 PCT~S94/02283 ~157~12 - 186 -t+) -anti-3- ~ 4-hydroxybenzamido) -~- [ 3, 5-dihy~roxy-~- ~ 2 -hydroxynaphthyl) carbonyl]benzoyloxyazepine trifluoroacetic acid salt ~COMPOIJND 511) To (+)anti-3-(4-benzyloxybenzamido)-4-[3,5-dibenzyloxy -4-(2-benzyloxynaphthyl)carbonyl] benzoyloxy-N-benzylazepine (338 mg, 0.366 mmol) dissolved in absolute ethanol (18 ml) under an atmosphere of nitrogen was added trifluoracetic acid (30 ~1, 0.386 mmol) followed by Pearlman's catalyst (135mg, 40 % by wt). An atmosphere of hydrogen was introduced and the mixture was allowed to stir for 48 h. The catalyst was removed by filtration and the volatiles removed under reduced pressure.
The product was chromatographed on a Dynamax-60 C18 column (41.4 mm ID X 30 cm length) using a linear gradient from 100%
A (0.1% TFA and 5% acetonitrile in water) to 50% B (pure acetonitrile) over 60 m at 25 ml/min. The product elutes in 50 minutes. Removal of the volatiles provided Compound 511 as a yellow solid (99 mg, 38%), mp 165-168-C. IR KBr (disc) cm 3397, 3274, 3121, 2874, 1796, 1776, 1680, 1633, 1606, 1544, 1510, 1461, 1426, 1369, 1344, 1202, 1142, 1109, 1054, 986, 910, 827, 802, 762, 723, 671. Anal. Calcd for C3lH28N20, 2CF3C02H:
C, 54.69; H, 3.93: N, 3.64. Found: C, 54.46; H, 4.06; N, 3.65.
W094l20062 215 7 412 PCT~S94/02283 ~rans-4-(~-~2-Ca Loh~ 6-hy~roxybenzoyl)-3,5-dihy~roxybenzoylo xy)-3-(~-hydroxybenzam~do)-1-(phenylaminocarbonyl)azepine (COMPOUND 512) HO~ HO~
(~. N~_ OH ~~, NH~_ OH
O NH
A solution of 25.2 mg (0.035 mmol) of (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)azepine trifluoroacetic acid salt in 0.30 ml of dry pyridine was treated with 6.5 ~L (7.1 mg, 0.060 mmol) of phenylisocyanate. The mixture was stirred for 3 h at room temperature, after which the reaction was quenched by the addition of 0.5 ml of methanol. The mixture was evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-100% B over 60 min, flow: 15 ml/min).
The pure fractions were pooled and evaporated and then lyophilized from water to give 23.8 mg (95%) of Compound 512 as W094/20062 - 188 - PCT~S94/022~
a yellow fluffy solid, mp >200 C. IR (KBr) : 1705, 1635, 1545, 1506, 1363, 1240, 760 cm . Anal. Calcd for C35H3lN3O11 -2.5 H2O: C, 58.82 H, 5.08; N, 5.88. Found: C, 59.04; H, 4.94;
N, 5.77.
WOg4/20062 21~ 7 412 PCT~S94tO2283 (+)-~nti-3-~-hy~roxybenzami~o)-~-t3,5-~ihy~roxy-~-(2,3,5,6-tetramethylphenyl)carbonyl]benzoyloxyazep~ne trifluoro~cetic ~cid salt (CONPOUND 513) To(+)-trans-3-(4-benzyloxybenzamido)-4-[3,5-~ihen7yl-oxy-4-(2,3,5,6-tetramethylphenyl)carbonyl]benzoyloxy-N-benzyl-azepine (330 mg, 0.364 mmol) dissolved in 1:1 absolute ethanol:ethyl acetate (100 ml) under an atmosphere of nitrogen was added trifluoracetic acid (42 ~1, 0.546 mmol) followed by Pearlman's catalyst (66 mg, 20 % by wt). An atmosphere of hydrogen was introduced and the mixture was allowed to stir for 24 h. The catalyst was removed by filtration and the volatiles were removed under reduced pressure. The product was chromatographed on a Dynamax~-60 C18 column (41.4 mm ID X 30 cm length) using a linear gradient from 100% A (0.1% TFA and 5%
acetonitrile in water) to 50% B (pure acetonitrile) over 60 m at 25 ml/min. The product elutes in 58 minutes. Removal of the volatiles under reduced pressure provided Compound 513 as a yellow solid (185 mg, 75%), mp 157-160-C. IR KBr (disc) cm 3084, 1717, 1682, 1637, 1609, 1559, 1542, 1509, 1474, 1457, 1425, 1375, 1339, 1270, 1234, 1199, 1140, 1108, 1076, 1007, 938, 848, 813, 767, 723, 669. Anal. Calcd for C3lH28N207 -CF3COzH H20: C, 58.40; H, 5.50; N, 4.13. Found: C, 58.26;
H, 5.28; N, 4.03.
wos4l2~K2 PCT~S94/02283 7 157 412 - lgo -tl)-Anti-4-t3~5-~imethoxy-4-~2~6-~ihydroxyphenylcarbonyl)]-3 (4-hydro~ybenz~mi~o)perhydroazepine (COMPOUND 514) H3CO~¢~OCH3 BuU & o~CH~`C02+ o~ OCH~ ~+ Ht~OOH
CO2+
&OBn O~CH CO~H ~ (~N~ o ~d OBn H2 ~ C~O
OH OCH3 ~- N~ OH
H
The above compounds were synthesized using reactions indicated. Compound 514 was isolated as a yellow powder (37 mg, 5.2% overall yield): mp 165-175 C; NMR (D6 DMSO); IR (KBr) cm : 3431 (OH); 1710 (ester); 1625 (ketone). Anal. calc. for C2~H3oN2~-l-5 H2O: C, 60.31; H, 5.76; N, 4.85. Found: C, 60.53;
H, 5.85; N, 4.91. Intermediate Compound 603 was also isolated.
_ W094/2~62 21~ 7 412 PCT~S9410~
syn-4- ( ~- ( 2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyl-amino)hexahy~ro-3-(4-hydroxybenzoylamino)azepine, trifluoro-acetic aci~ salt (COMPOUND 516) Hexahydro-3-(~-phenylmethoxy)benzoyl~mino-1-phenylmethyl-~zepin ~-one ~ A 25 ml 3-neck round bottom flask under nitrogen was charged with 2.0 N oxalyl chloride/methylene chloride (Aldrich, 1.125 ml, 2.25 mmol), diluted with anhydrous methylene chloride (2 ml), cooled (-65-C), and treated dropwise with anhydrous dimethylsulfoxide (0.35 g, 4.5 mmol) in anhydrous methylene chloride (1.2 ml) at a rate to keep the pot temperature below -60-C. The mixture was stirred at -65 + 5C for 30 min, then treated dropwise with a solution of syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.645 g, 1.5 mmol) in anhydrous methylene chloride (1.5 ml) at a rate to keep the pot temperature below -55-C. The mixture was stirred at 55+5-C for 2 h, then treated dropwise with triethylamine (1.5 ml), warmed to room temperature over one hour, and diluted with methylene chloride (10 ml). The organic solution was washed with water (10 ml), saturated aqueous sodium bicarbonate (10 ml), dried (Na2SO4), and concentrated in vacuo. The residue was chromatographed on silica gel (eluted with 5% acetone/
methylene chloride) to afford hexahydro-3-(4-phenyl-methoxy) benzoylamino-l-phenylmethylazepin-4-one (0.53 g, 82%) as a viscous colorless oil.
~exahydro-3-(~-phenylmethoxy)benzoylamino-1-phenylmethyl-azepin-~-one, oxime A solution of hexahydro-3-(4-phenylmethoxy)-benzoylamino-l-phenylmethyl-azepin-4-one (0.87 g, 2.03 mmol) in ethanol (12 ml) was treated with hydroxylamine hydrochloride (0.19 g, 2.73 mmol), followed by 25% methanolic sodium methoxide (Aldrich, 0.20 g, 0.93 mmol), and was heated to 50C
for one hour. The mixture was cooled to room temperature and treated with additional 25~ methanolic sodium methoxide (0.42 g, 1.94 mmol), then concentrated in vacuo to afford hexahydro-WOg4/2~62 PCT~S94/022~
21S7~12 - 192 -3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepin-4-one oxime (0.89 g, 99%) as a colorless foam.
syn-4-(3,5-Bis~phenylmethoxy)-4-~2-ca~hophenylmethoxy-6-phenyl-methoxybenzoyl)benzoylaminohexahydro-3-~4-phenylmethoxy)-benzoyl~mino-1-phenylmethylazepine ~COMPOUND 615) A solution of hexahydro-3-(4-phenylmethoxy)-benzoyl amino-l-phenylmethyl-azepin-4-one oxime (0.40 g, 0.90 mmol) in reagent methanol (25 ml) in a Parr bottle was treated with Raney Nickel (Aldrich, quarter tsp.), then subjected to hydrogenation at 49-50 psi for six hours. The solution was carefully evacuated of hydrogen, filtered through celite, and the filtrate was concentrated in vacuo to afford 4-aminohexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenyl methylazepine, 1:1 mixture of isomers, which was kept-under nitrogen. Meanwhile, 2'-carbobenzyloxy-2,6,6'-tribenzyloxy benzophenone-4-carboxylic acid (SPC-104034, b.37 g, 0.S5 mmol) was placed in a round-bottom flask and repeatedly covered with toluene and concentrated in vacuo to remove all water and other persistent solvents. Finally, the residue was dissolved in anhydrous methylene chloride (2 ml) under nitrogen, treated with dimethylformamide (3 drops), then with 2.0 N oxalyl chloride/methylene chloride (0.4 ml, 0.8 mmol), and stirred at room temperature for one hour. The solution was concentrated in vacuo, placed under high vacuum for one hour, then dissolved in methylene chloride (3 ml) and added to the 4-aminohexahydro-3-(4-phenylmethoxy)-benzoylamino-1-phenylmethylazepine prepared above. Sodium hydroxide (1.0 N, 1.5 ml) was added, and the mixture was stirred for one hour and separated. The aqueous layer was extracted with methylene chloride (2x10 ml), and the combined organic layer and extracts were washed with saturated sodium chloride (10 ml), dried (Na2SO~), and concentrated in vacuo. The residue was chromatographed (flash) on silica gel (eluted successively with 3% acetone/methylene chloride, 5%
acetone/methylene chloride, and 8% acetone/methylene chloride) to afford, initially, syn-4- ( 3,s-bis(phenylmethoxy)-4- ( 2-carbophenylmethoxy-6-phenylmethoxybenzoyl)-benzoylamino _ V094/2~ 2 1 ~ 7 4 1 2 PCT~S94/0~
hexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenylmethyl-azepine (0.26 g, 43%), then anti-4-(3,5-bis(phenylmethoxy)-4-(2-carbophenyl-methoxy-6-phenylmethoxybenzoyl)benzoylamino hexahydro-3-(4-phenyl-methoxy)benzoylamino-1-phenylmethyl azepine (0.21 g, 35%) as colorless foams. The combined yield was 0.47 g (78%).
syn~ - (2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyl-~m~no)hexahydro-3-(~-hy~roxybenzoyl~m~no)azepine, trifluoro-acetic acid salt (CO~PO~ND 516) A solution of anti-4-(3,5-bis(phenylmethoxy)-4-(2-carbophenylmethoxy-6-phenylmethoxybenzoyl)benzoylaminohexahydro -3-(4-phenylmethoxy)-benzoylamino-1-phenylmethylazepine (0.20 g, 0.183 mmQl) in reagent ethanol (9 ml) and ethyl acetate (1 ml) in a 2-neck 25-ml round bottom flask under nitrogen was treated with Pearlman's catalyst (20% Pd(OH)2/C, 50 mg) and trifluoroacetic acid (42 mg, 0.37 mmol). The flask was fitted with a balloon and a balloon valve, purged with hydrogen, and placed under positive hydrogen pressure for 18 h, then evacuated of hydrogen and purged for several minutes with nitrogen. The solution was carefully filtered through celite (wash filter pad with ethanol) and the filtrate was concentrated in vacuo to a yellow foam. This was dissolved in methanol (20 ml), diluted with deionized water (60 ml), and concentrated in vacuo to remove the methanol. Freeze drying (withoutchromatography)affordedsyn-4-(4-(2-carboxy-6-hydroxy benzoyl)-3,5-dihydroxybenzoylamino)hexahydro-3-(4-hydroxy benzoylamino)azepine (94 mg, 68%) as a voluminous yellow solid;
mp >300-C(dec). Rf (4% acetic acid/ethanol) 0.50; IR (KBr) 1683, 1636, 1604 cm1; 1H NMR (d6-DMS0) ~ 11.66 (s, 2H), 10.07 (s, lH), 9.86 (s, lH), 8.60 - 9.00 (br s, 2H), 8.33 (d, lH, J
= 8 Hz), 8.00 (d, lH, J = 7 Hz), 7.70 (d, 2H, J = 9 Hz), 7.37 (d, lH, J = 8 Hz), 7.27 (t, lH, J = 8 Hz), 7.06 (d, lH, J = 8 Hz), 6.82 (d, 2H, J = 9 Hz), 6.68 (s, 2H), 4.55 (m, lH), 4.40 (m, lH), 3.10 - 3.50 (m, 4H), 1.70 - 2.10 (m, 4H). Anal.
Calcd. for C2~H27N30~-l.5(C2H02F3)-2.0(H20): C, 49.21; H, 4.33; N, 5.55. Found: C, 48.82; H, 4.59; N, 5.79.
2 1 5 ~ 4 1 ~ - 194 - PCT/US94/02283 syn- ~ - t ~ - ( 2 -hydroxycarbonyl- 6-hy~roxybenzoyl ) - 3, 5-0 ihydroxy benzoyloxy] -3- (~-hy~roxybenzamido) perhydroazepine trifluoroacetic aci~ salt (CONPO~rND 517 ) BnO2C O OBn BnO2C O OBn OH OBn O~ COCI ~ H OBn ~,H~ Et3N, DMAP. CH2C12 (~Nb~
Bn Bn 1. Pd(OH)2-C H ~ ~OH 610 EtOH. EtOAc OH OH ~, O
COMPO~lND 610 To a chilled solution (0-5 C) of 4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoic acid (0.52 mmol, 350 mg) in 2 ml of methylene chloride under nitrogen atmosphere was added oxalyl chloride (0.75 mmol, 95 mg) in one portion. N,N-Dimethylformamide (1 drop) was added and the light brown solution was stirred at 0 C for 1.5 h. The solvent and excess oxalyl chloride were then removed in vacuo, the resulting brown oil was redissolved in methylene chloride (2 ml) and added to a cooled solution (0-5 C) of syn-N-benzyloxycarbonyl -3 - ( 4 -hydroxybenzamido ) -4 -hydroxyperhydroazepine (0.62 mmol, 226 mg), triethylamine (1.29 mmol, 130 mg) and 4-dimethylaminopyridine (approximately 10 mg) in 2 ml methylene chloride (nitrogen atmosphere). The reaction mixture was allowed to warm to room temperature and ~0 94l20062 2 1 !~ 7 ~ 1 2 PCT/US94/02283 stirred overnight (approximately 18 h) under a nitrogen atmosphere, after which it was diluted with 50 ml of methylene chloride, washed with saturated sodium bicarbonate solution (lo ml), water (10 ml), and brine (10 ml), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give 440 mg of the crude product. Chromatography on silica gel eluting with 4:1-hexane:ethyl acetate gave 399 mg (71%) of the coupled product as a light yellow solid, which was used directly in the next step.
To a solution of syn-N-benzyloxycarbonyl-4-[4-(2-benzyloxycarbonyl-6-benzyloxybenzoyl) -3, 5-dibenzyloxy benzoyloxy]-3-(4-benzyloxybenzamido)perhydroazepine (0.16 mmol, 180 mg) in ethyl acetate (5 ml) and absolute ethanol (10 ml) was added moist 20% palladium hydroxide on carbon (20% w/w, 36 mg). The reaction flask was fitted with a hydrogen balloon and the grey suspension was stirred at room temperature for 18 hours. The reaction flas~c was then purged with nitrogen gas and the solution diluted with chloroform (50 ml), filtered over celite, treated with 1 ml of trifluoroacetic acid, and concentrated in vacuo to give 87 mg of the crude product. The material was purified by HPLC chromatography with a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA;
solvent B: 100% acetonitrile; gradient 0-50% B over 60 minutes, flow: 15 ml/min). The purified fractions were concentrated and lyophilized from water to give 67 mg (63%) of the title compound as a yellow fluffy solid. IR (KBr): 1704, 1688, 1677, 1632, 1608, 1427, 1235, 1201 cm 1; EA (calculated for C28H26N2Ol0-1.2 C2HF3O2-2.0 H2O): C, 50.48; H, 4.35 N, 3.87.
Found: C, 50.51; H, 4.46; N, 3.88.
W094/2 62 - 196 - PCT~S94/0~
I+)-anti-3-t4-hydroxybenzamido)-4-t3,5-dimethoxy-~-(2,6-dimethoxy)benzoyl]benzoyloxylperhydroazepine trifluoroAcetic aci~ salt (COMPOUND 518) To (+)-anti-3-(4-benzyloxybenzamido)-4-t3,5-dimethoxy-4-(2,6-dimethoxy)benzoyl]benzoyl~oxy-N-benzylperhydroazepine (221 mg, 0.291 mmol) dissolved in ethyl acetate (50 ml) under an atmosphere of nitrogen was added trifluoracetic acid (35 ~L, 0.437 mmol) followed by Pearlman's catalyst (44 mg, 20 % by wt on carbon). An atmosphere of hydrogen was introduced and the mixture allowed to stir for 48 h. The catalyst was removed by filtration and the volatiles were removed under reduced pressure. The product was chromatographed on a Dynamax~-60 C18 column (41.4 mm ID X 30 cm length) using a linear gradient from 100% A (0.1% TFA and 5% acetonitrile in water) to 100% B (pure acetonitrile) over 60 m at 25 ml/min. The product elutes in 58 minutes. Removal of the volatiles under reduced pressure provided Compound 518 as a white solid (62 mg, 26%), mp 134-137-C. IR KBr (disc) cml 3430, 3105, 3016, 2948, 2843, 1772, 1677, 1592, 1544, 1508, 1474, 1436, 1411, 1331, 1235, 1203, 1182, 1127, 1111, 950, 917, 848, 798, 766, 720, 620, 602.
Anal. Calcd for C31H34N2Og 2CF3CO2H: C, 52.11; H, 4.50; N, 3.47. Found: C, 52.30; H, 4.50; N, 3.47.
(+)-~nti-3-(~-benzyloxybenzami~o)-~-t3,5-dimethoxy-~-(2,6-dimethoxy)benzoyl]benzoyloxy-N-benzylperhydroazepine ~COMr~uN~ 60~) To a solution of 3,5-dimethoxy-4-(2,6-dimethoxybenzoyl)benzoic acid (221 mg, 0.639 mmol) in anhydrous dichloromethane (10 ml) under an atmosphere of nitrogen at 0-C
was added oxalyl chloride (436 ~L, 2 M in dichloromethane, 0.872 mmol) dropwise over 5 minutes followed by anhydrous dimethylformamide (3 drops). The ice bath was removed and the suspension rapidly turned into a clear yellow solution. The reaction mixture was allowed to stir for 0.5 h at room temperature. The volatiles were removed under reduced pressure and the remaining solid was dried under vacuum for 2.5 h.
_ ~094/2~K2 2 ~ 5 7 412 pcT~ss4lo22~
To a solution of (+)-anti-3-(4-benzyloxybenzamido)-4-hydroxy-N-benzylperhydroazepine (250 mg, 0.581 mmol), triethylamine (267 ~L, 1.92 mmol), and dimethylaminopyridine (7.1 mg, 0.0581 mmol) in anhydrous dichloromethane (10 ml) under an atmosphere of nitrogen at 0C was added a solution of the above generated acid chloride in anhydrous dichloromethane (10 ml) dropwise over 0.5 h. After allowing to stir while warming to room temperature overnight the reaction mixture was diluted with dichloromethane (200 ml) and washed with water (75 ml). The dichloromethane layer was dried over magnesium sulfate, filtered, and the volatiles were removed under reduced pressure to give a crude white solid. The solid was purified using flash column chromatography (silica gel, 9 : 1 dichloromethane and ethyl acetate) to provide Compound 604 as a white solid (266 mg, 60%), mp 82-85-C. IR KBr (disc) cm 3481, 3029, 2939, 2837, 1711, 1676, 1644, 1591, 1530, 1498, 1474, 1407, 1327, 1248, 1177, 1111, 1024, 995, 913, 845, 795, 744, 700, 605. Anal. Calcd for C45H46N20~: C, 71.22; H, 6.11;
N, 3.69. Found: C, 71.08; H, 6.15; N, 3.64.
3,5-dimethoxy-4-(2,6-dimQthoxy)bQnzoylbenzoiG acid To t-butyl-3,5-dimethoxy-4-(2,6-dimethoxybenzoyl) benzoate (2.26 g, 5.62 mmol) under an atmosphere of nitrogen at O C was added formic acid (25 ml) dropwise. The ice bath was removed and the reaction mixture was allowed to stir at room temperature for 8 h. The volatiles were removed under reduced pressure. Recrystallization of the crude solid with 1:1 ethyl acetate:hexanes provided the title compound as a white solid (1.69 g , 87%), mp 209-212-C. H NMR (DMSO-d6) ~ 7.32 (t, 1 H, J = 8 Hz), 7.16 (s, 2 H), 6.64 (d, 2 H, J = 8 Hz), 3.67 (s, 6 H), 3.61 (s, 6 H); IR KBr (disc) cm1 3432, 3199, 3103, 3016, 2936, 2838, 1731, 1653, 1589, 1475, 1429, 1409, 1313, 1258, 1222, 1185, 1128, 1113, 1028, 944, 897, 869, 801, 773, 713, 690, 617. Anal. Calcd for C1~Hl~O7: C, 62.42; H, 5.24. Found:
C, 62.43; H, 5.30.
t-butyl-3,5-dimethoxy-4-(2,6-dimQthoxybenzoyl)benzoatQ
W094/2~62 PCT~S94/0~
2157 ~ 198 -To a solution of t-butyl-3,5-dimethoxy-4-[(2,6-dimethoxyphenyl)hydroxymethyl]benzoate (3.10 g, 7.70 mmol) in acetone (50 ml) at O C was added Jones reagent dropwise until the reaction mixture retained the orange color of the Jones reagent. The reaction mixture was diluted with dichloromethane (500 ml) and washed with water (150 ml). The dichloromethane layer was dried over anhydrous magnesium sulfate, filtered and the volatiles were removed under reduced pressure. The crude residue was purified using flash column chromatography (silica gel, 1 : 8 ethyl acetate / hexane) to provide the title compound as white solid (2.36 g, 88%), mp 54-57-C. lH NMR
(DMSO-d6) ~ 7.32 (t, 1 H, J = 8.5 Hz), 7.11 (s, 2 H), 6.63 (d, 2 H, J = 8.5 Hz), 3.67 (s, 6 H), 3.61 (s, 6 H) 1.55 (s, 9 H);
IR KBr (disc) cm1 2974, 2839, 1711, 1685, 1590, 1473, 1434, 1406, 1369, 1330, 1292, 1253, 1163, 1126, 1111, 1032, 957, 915, 849, 796, 765, 705, 669, 610. Anal. Calcd for C22H26O7: C, 65.66; H, 6.51. Found: C, 65.67; H, 6.49.
t-butyl-3,5-dimethoxy-~-~(2,6-dimethoxyphenyl)hydroxy-methyl]benzoate Toasolutionoft-butyl-3,5-dimethoxy-4-bromobenzoate (3.50 g, 11.0 mmol) in anhydrous tetrahydrofuran (60 ml) under an atmosphere of nitrogen with an internal temperature of -78-C
(ether/dry ice) was added n-butyllithium (7.58 ml, 1.6 M in hexanes, 12.1 mmol) dropwise at a rate which did not allow the internal temperature to rise above -65-C. To the reaction mixture was added a solution of 2,6-dimethoxybenzaldeyde (1.83 g, 12.1 mmol) in anhydrous tetrahydrofuran (20 ml) dropwise at a rate which did not allow the internal temperature to rise above -65-C. The reaction mixture was allowed to stir while warming to room temperature over 2 h. The reaction mixture was quenched with solid ammonium chloride and the volatiles were removed under reduced pressure. The crude residue was diluted with ethyl acetate (500 ml) and washed with water (200 ml).
The ethyl acetate layer was dried over anhydrous magnesium sulfate, filtered, and the volatiles were removed under reduced pressure. The crude residue was purified by recrystallization W094/2~62 215 ~ 412 PCT~S94/0~
from 1 : 1 ethyl acetate : hexane which provided a white solid of the title compound (3.2 g, 72%), mp 131-133-C. H NMR
(DMSO-d6) ~ 7.14 (t, 1 H, J = 8.5 Hz), 7.08 (s, 2 H), 6.58 (d, 2 H, J = 8.5 Hz), 6.43 (d, 1 H, J = 10.5 Hz), 5.37 (d, 1 H, J
= 10.5 Hz) 3.76 (s, 6 H), 3.70 (s, 6 H) 1.63 (s, 9 H);IR KBr (disc) cml 3541, 3445, 2978, 2843, 1701, 1651, 1588, 1542, 1477, 1455, 1417, 1366, 1328, 1241, 1161, 1119, 1030, 958, 847, 809, 770, 667. Anal. Calcd for Cz2H2~O7: C, 65.33; H, 6.98.
Found: C, 65.19; H, 6.99.
t-butyl-~-bromo-3,5-~imethoxybenzoate To a solution of 4-bromo-3,5-dimethoxybenzoic acid (9.20 g, 35.2 mmol) in anhydrous dimethylformamide (200 ml) under an atmosphere of nitrogen was added N,N-carbonyl diimidazole (6.29 mg, 38.8 mmol). After the reaction mixture was allowed to stir for 1 h at room temperature, DBU (5.80 ml, 38.8 mmol), and t-butanol (9.97 ml, 106 mmol) were added. The reaction mixture was heated at 80-C for 2 h. The reaction mixture was quenched by the slow addition of water (300 ml).
The solid which formed was collected by suction filtration and washed with water (3 X 30 ml) which provided the title compound as a white solid (5.8 g, 52%), mp 119-121-C. lH NMR (DMSO-d6) ~ 7.17 (s, 2 H), 3.89 (s, 6 H), 1.57 (s, 9 H); IR KBr (disc) cm-l 2979, 2936, 2836, 1708, 1590, 1456, 1408, 1366, 1337, 1258, 1231, 1174, 1124, 1033, 960, 8S7, 798, 761, 643. Anal.
Calcd for C13Hl7BrO4: C, 49.23; H, 5.40. Found: C, 49.05; H, S.36.
~-bromo-3,5-dimethoxybenzoic acid To methyl 4-bromo-3,5-dimethoxybenzoate (10 g, 36.4 mmol, Pharmatech International) was added sodium hydroxide (75 ml, 10 N) and methanol (50 ml). The reaction mixture was heated at 70-C for l.S h. The reaction mixture was cooled to Q-C and slowly acidified using HCl (6N). The solid was collected by suction filtration to provide a white solid of the title compound (9.31 g, 98%), mp 22S-228-C. 1H NMR (DMSO-d6) ~ 13.3 (br s, 1 H), 7.23 (s, 2 H), 3.90 (s, 6 H); IR KBr ~ 5 7 4 1~ PCT~S94/022~ _ (disc) cml 3400, 3064, 2969, 2838, 2745, 1689, 1586, 1461, 1407, 1329, 1276, 1230~, 1189, 1127, 1039, 935, 857, 764, 730, 667, 639. Anal. Calcd for CgHgBr04: C, 41.41; H, 3.47. Found:
C, 41.44; H, 3.40.
_ W094/20~2 2 1 ~ 7 412 PCT~S94/02283 ~3R,~R)-[~-(2,6-Dimethoxybenzoyl)-3,s-~ihy~roxybenzoyloxy~-3-(~-hy~roxybenz~mido)perhy~ro~zepine trifluoro~cetic ~cid ~lt (COMPOUND 519) Br BnO~OBn1)gULi,THF,-72C OMe O ~
~Me ~CO2t-Bu CO2t-Bu2) ~ OMe OBn OMe Jones Reagent OMeo OMe o ~COOH , HCOOH ~ CO2t-Bu OMe OBn OMe OBn 1) (cOc1)2 ,OH
~) (;;~Nb~OBn ~ Ph PH320~7A
Et3N, DMAP
0~ 0~
O BnO 1)TFA
~S--HN ,~OBn 2) H2 ~tNb~O H
~NJ O PdoH)2 / C ~NJ O
- Fh TFA
WO94t2~62 215 7 ~12 PCT~S94/0~
l,l-Dimethylethyl 3,5-dibenzyloxy-~-[~2,6-dimethoxyphenyl)-hydro~ymethyl) benzoate 2.5 ml (7.02 mmole) of 2.5 M n-butyllithium in hexane was added dropwise to a cold solution (-72-C) of t-butyl-4-bromo-3,5-dibenzyloxybenzoate (2.69 g, 5.47 mmole) in 40 ml of anhydrous tetrahydrofuran (THF) under nitrogen. The temperature of the solution was maintained below -70-C
throughout the addition and the solution was stirred in the cold for ten minutes. A solution of 1 g (6.02 mmole) of 2,6-dimethoxybenzaldehyde in 15 ml of THF was added dropwise maintaining the temperature below -65-C throughout the addition. The solution was allowed to warm to room temperature as it stirred for two hours. The solution was partitioned between 20 ml of 1 N hydrocholoric acid and 50 ml of ethyl acetate while stirring fifteen minutes. The organic layer was separated, washed with water, saturated brine and dried over magnesium sulfate. The solvent was removed in vacuo. The residue was recrystallized from ethyl acetate-hexane to give 0.83 g (26%) of the title compound as white crystals, mp 152-155-C. Anal. Calcd for C34H36O7: C, 73.36; H, 6.51. Found:
C, 73.04; H, 6.54.
l,1-Dimethylethyl 4-(2,6-dimethoxybenzoyl)-3,5-dibenzoyl oxybenzoate To a cold solution (O-C) of 0.7 g (1.25 mmole) of 1,1-dimethylethyl 3,5-dibenzyloxy-4-[(2,6-dimethoxyphenyl)-hydroxymethyl) benzoatein 15 ml of acetone was added 4 ml of Jones reagent. The solution was stirred in the cold for two hours. To the solution was added 10 ml of isopropyl alcohol to destroy the Jones reagent. The reaction mixture was filtered through celite and washed through with acetone. The filtrate was concentrated in vacuo. The residue was recrystallized from ethanol-water to yield 0.47 g (68%) of the title compound as tan crystals. Anal. Calcd for C34H3407: C, 73.63: H, 6.18.
Found: C, 73.15; H, 6.38.
WO94/2~K2 21~ 7 412 PCT~S94/022~
~-(Z,6-Dimethoxybenzoyl)-3,5-~ibenzyloxybenzoic acid A solution of 0.46 g (0.83 mmole) of l,l-dimethylethyl 4-(2,6-dimethoxybenzoyl)-3,5-dibenzyloxybenzoate in 10 ml of formic acid was stirred at room temperature for two hours.
After stirring for one hour a precipitate formed. The reaction mixture was poured over ice water, and the resultant precipitate was collected and dried to yield 0.36 g (87%) of a tan solid.
Trans-N-Benzyl-4-(~-(2,6-dimethoxybenzoyl)-3,5-~;~^n~yloxy)-3-(4-benzyloxybenz~mido)~zepine (COMPOUND 605) A solution of 0.36 g (0.72 mmole) of 4-(2,6-dimethoxybenzoyl)-3,5-dibenzyloxybenzoic acid in 10 ml of methylene chloride containing a trace (approximately 1 ~L) of dimethylformamide was cooled to 0-C. A 2.0 M solution of oxalyl chloride in methylene chloride (0.41 ml, 0.82 mmole) was added, and the mixture was stirred under a nitrogen atmosphere for ninty minutes. The reaction mixture was evaporated and the residue was evaporated twice from 15 ml of methylene chloride.
The residue was dissolved in 8 ml of methylene chloride and was added to a solution of 0.35 g (0.82 mmole) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine, 0.06 ml (0.82 mmole) triethylamine, and 4.0 mg of DMAP in 10 ml of methylene chloride. The solution was stirred at room temperature under nitrogen for sixteen hours. The solution was diluted with 30 ml of methylene chloride and washed with saturated sodium bicarbonate, saturated brine, dried over magnesium sulfate, and the solvent was removed in vacuo. The residue was chromatographed on silica gel eluting with hexane-ethyl acetate (70:30). Yield 0.33 g (43%) of a glassy oil which solidified on standing.
(3R,~R)-t~-(2,6-Dimethoxybenzoyl)-3,5-dihy~roxybenzoyloxy]-3-(~-hydroxybenzamido)perhydroazQpine trifluoroacetic acid salt (COMPO~ND 519) A solution of 0.33 g (0.36 mmole) of trans-N-benzyl-4-(4-(2,6-dimethoxybenzoyl)-3,5-dibenzoyloxy)-3-(4-benzyloxy W094/20062 - 204 - PCT~S94/02283 benzamido)azepine in 20 ml ethanol-methylene chloride (1:1) was treated with 0.055 ml (0.720 mmole) of trifluoroacetic acid.
The solution was stirred for five minutes. The solvent was evaporated. The ethanol-methylene chloride solvent was added twice more and evacuated in order to remove excess trifluoroacetic acid. The residue was taken up in 15 ml of ethanol, cooled to O-C, and 0.5 g of moist 10% palladium hydroxide on carbon was added. The mixture was then stirred under an atmosphere of hydrogen for six hours at room temperature. The mixture was filtered, evaporated, and the residue was chromatographed on a 41 X 250 mm C 18 column (solvent A: 95 : 5 water / acetonitrile + 0.1% TFA; solvent B:
100% acetonitrile; gradient : 0 - 50% B over 60 min., flow: 25 ml / min.). The pure fractions were pooled and evaporated to yield 0.158 g (60%) of Compound 519, a yellow powder, mp 189-193 C. IR (KBr); 1678, 1605, 1508, 1427, 1370, 1250, 1200, 765 cml. Anal. Calcd for C29H30N2O~ 2H2O B 1-.3 TFA: C, 51.65;
H, 4.84: N, 3.81. Found: C, 51.27; H, 4.68; N, 3.62.
Trans-4- ~4- (2-cis-carboxycyclohexylcarbonyl) -3, 5-~ihydroxybenzoyloxy) -3 - ( 4 -hy~roxybenzamido ) ~zepine trifluoroacetic ~cid salt (COMPO~rND 519) HOOC O OBn BnO~,OBn l)BULi. THF, -72Qc [~C02t Bu CO2t-Bu 2) ~O
BnBr DMF
BnOOC O OBn BnOOC O OBn [J~ , HCOOH ~¢,~
BnO CO2H BnOCO2t-Bu 1) (cOc1)2 ~O H
2) ( )~...N~OBn N O
~Ph ~OBn 1)TFA 0~
(~... N~OBnPd(OH)2 / C (~S~N~O H
- ~ Fh TFA
1, 1-Dimethylethyl 3, 5-dibenzyloxy-4- (2 -Ci9-carboxycyclohexylcarbonyl ) benzoate 2 . 8 ml (7.03 mmole) of 2.5 N n-butyllithium in hexane was added dropwise to a cold solution (-72 C) of 3.0 g (6.39 mmole) of 1,1-dimethylethyl 4-bromo-3,5-dibenzyloxybenzoate in W094/2~62 Z 15 ~ 412 PCT~S94/0~
40 ml of anhydrous tetrahydrofuran (THF) under nitrogen. The solution was stirred in the cold for ten minutes. A solution of 1.2 g (7.03 mmole) of cis-1,2-cyclohexanedicarboxylic anhydride in 10 ml of THF was added dropwise maintaining the temperature below -70 C. The solution was stirred in the cold for two hours. The solution was poured into 150 ml of saturated ammonium chloride and 350 ml of ether. The reaction mixture was stirred for thirty minutes. The organic layer was separated, washed with 0.1 N hydrochloric acid, saturated brine and dried over magnesium sulfate. The solvent was evaporated to yield 3.4 g (98 % ) of a clear oil.
1,1-Dimethylethyl ~ 2-cis-benzyloxycyclohexylcarbonyl))-3,5-dibenzyloxybenzoate To a solution of 3.40 g (6.20 mmole) of 1,1-d im e t h y l e t h y l 3, 5 - d i b e n z y l o xy -4 - (2 - c i s -carboxycyclohexylcarbonyl)benzoate in 20 ml of dry DMF was added 0.43 g (3.10 mmole) of potassium carbonate and 0.33 ml (3.10 mmole) of benzyl bromide. The solution was stirred at room temperature under nitrogen for eight hours. Starting material was still present in the reaction. Therefore, an additional 0.16 ml (1.5 mmole) of benzyl bromide was added and stirring was continued for sixteen hours. The solution was poured over 100 ml of ice water, extracted twice with 150 ml portions of ether. The ether solution was washed with water, saturated brine and dried over magnesium sulfate. The solvent was evaporated and the residue was chromatographed on silica gel eluting with 5 % - 10 % ethyl acetate - hexane to yield 1.0 g (25 %) of a clear oil.
4-(4-~2-cis-benzyloxycyclohQxylc~rbonyl)-3,5-~ih~nzyloxybenzoic ~ci~
A solution of 0.45 g (0.71 mmole) of 1,1-dimethylethyl 4-(4-(2-cis-benzyloxycyclohexylcarbonyl)-3,5-dibenzyloxybenzoate in 10 ml of formic acid was stirred at room temperature for four hours. The solution was poured over ice water, the resultant precipitate was collected and dried to yield 0.34 g (82.8%) of a white solid. Anal. Calcd for C36H34O7 : C, 74.72 ; H, 5.92. Found : C, 74.46 ; H,5.99.
wo g4t20062 2 1 ~ ~ 4 1 2 PCT~S94/0 Trans-N-benzyl-4-(4-~2-cis-bQnzyloxycyclobexylcarbonyl)-3-~4-benzyloxybenz~mido)azepine ~COMPOUND 606) A solution of 0.40 g (0.69 mmole) of 4-(4-(2-cis-benzyloxycyclohexylcarbonyl)-3,5-dibenzyloxybenzoic acid in 8 ml of methylene chloride containing a trace (amount approximately 1 ~L) of dimethylformamide was cooled to o oc.
A 2.0 M solution of oxalyl chloride in methylene chloride (0.59 ml, 1.18 mmole) was added and the solution was stirred under nitrogen for 2.5 hours. The reaction mixture was evaporated, and the residue was evaporated twice from 15 ml of methylene chloride. The residue was dissolved in 8 ml of methylene chloride and added to a solution of 0.34 g (0.78 mmole) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine,0.08 ml (0.78 mmole) of triethylamine and 4.0 mg of DMAP in 10 ml of methylene chloride. The solution was stirred at room temperature under nitrogen for sixteen hours. The solution was diluted with 30 ml of methylene chloride and washed with saturated sodium bicarbonate, saturated brine and dried over magnesium sulfate. The solvent was evaporated and the residue was chromatographed on silica gel eluting with ethyl acetate-hexane (1:4). Yield of 0.5 g (73%) of a clear oil.
Trans-4-~ 2-cis-carboxycyclohexylcarbonyl)-3,5-dihydroxybenzyloxy)-3-(~-hy~roxybenzamido) a zepine trifluoroacetic acid salt (CO~POUND 520) A solution of 0.38 g (0.38 mmole) of trans-N-benzyl-4-(4-(2-cis-benzyloxycyclohexylcarbonyl))-3-(4-benzyloxybenzamido)azepine in a 25 ml mixture of methanol, ethanol and methylene chloride (1 : 2 : 2 ) was treated with 0.06 ml (0.77 mmole) of trifluoroacetic acid for five minutes.
The solvent was evaporated, and the methanol, ethanol, methylene chloride solvent was added twice more and evaporated.
The residue was taken up in 15 ml of ethanol, cooled to 0 C
and 0.05 g of palladium hydroxide on carbon (20% by wt.) was added. The mixture was then stirred under an atmosphere of hydrogen for six hours at room temperature. The mixture was filtered, evaporated and the residue was chromatographed on a WO 94/20062 PCTtUS94tO2283 2157 ~ 208 -41 X 250 mm C18 column (solvent A: 95: 5 water / acetonitrile ~ 0.1% TFA; solvent B : 100% acetonitrile ; gradient O - 50% B
over 60 min., flow 25 ml / min.). The pure fractions were pooled to yield 57 mg (21%) of a yellow powder, mp 122 - 127 C. IR (KBr): 1676, 1607, 1508, 1427, 1365, 1276, 1203, 757 cm . Anal. Calcd for C2ôH32N209 2H20 1.2 TFA : C, 51.18 ; H, 5.25 ; N, 3.92. Found : C, 51.46 ; H, 5.37 ; 3.92.
_ N094/20062 2 15 7 412 PCT~S94/0~
~ Anti-~-t3~5-~ihy~roxy-~-(2~6-~ihy~.ohyLo~zoyl)]hexahy~ro-3 (~-hydroxybenzoyl~mine)~zepine (COMPOUND 521) OBn O OBn BnO ~ OBn OBn OBn OBn O OBn OBn O OBn HCOOH ~J~ (I)BnBr ~
OH OBn C02H (2)KOH OBn OBn CO2H
OH O OH
(I)Courlin~ ~
(2)H~ ~ ~
OH OH O CO ~ OH
,~
) 52 N~ H
tert-butyl 4-(2,6-dibenzyloxybenzoyl)-3,5-dibenzyloxybenzoate (360 mg, 0.51 mmol) was placed in formic acid(10 ml). The resulting suspension was stirred for 20 min and intermittantly heated with a heat gun. The reaction was poured over water (300 ml) and stirred. The solids that precipitated were then filtered. Next, the solids were dissolved in ethyl acetate, and dried over sodium sulfate. The sodium sulfate was filtered off, and the filtrate was concentrated in vacuo and recrystalized in hexane: ethyl acetate to yield a light yellow solid (Acid 175 mg). This ~ ~ 7 ~1 ~ PCT~S94/0~
material was dissolved in dimethylformamide (2 ml). Potassium carbonate (2.2 eq.) was then added. Next, benzyl bromide (5 eq) was added. The reaction was stirred at R.T. for 2 h. The reaction was taken up in ethyl acetate and lN HCl, and placed in a separatory funnel. The organic layer was isolated, dried over sodium sulfate, concentrated in vacuo, and flash chromatographed eluting with hexane:ethyl acetate / 90:10. The major product was isolated as a white foam. This material was then dissolved in a solution of methanol:isopropyl alcohol:water / 45:45:10 with 5% potassium hydroxide. The solution was stirred for 4 h at R.T. The organic solvents were then removed in vacuo, and to the remaining aqueous solution was acidified with lN HCl. Ethyl acetate (100 ml) was then added. The biphasic solution was then placed in a separatory funnel and the organic phase was isolated, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was recrystalized in hexane:ethyl acetate to yield a light yellow solid (92 mg). This benzophenone acid was then converted to Compound 521. Compound 521 was prepared through coupling and reduction in the same was as for Compound 547. A light yellow solid was obtained through reverse phase C-18 HPLC and subsequent lyophilization (7.5 mg). The structure was identified as Compound 521 through 1H NMR.
~094/2~K2 2 1 S 7 ~ 1 2 - 211 - PCT~S94/022~
~ anti-2-(~-~yd~hyL~hzamido)-3-t3,s-dihy~roxy-~-(2-hy~roxy-6-c~rboxyphenylcarbonyl)benzoyloxy]-N-benzylcaprolactam (COMPOUND 523) OBn 1. (COCI)2 cat. DMF, ~OBn B~OOC~L OBn HO N~ 2. Et3N, DMAP, BnO~ OBn =~ OB~ C~'F CH2CI2 ~O N
BnO~ N~_ S1% ~XF
o~OH ~-anti ~ ~-anti ~~ h HOOC ¢~L OH
O I~OH OH H2, Pd(OH)2 HO ~ H _~ 95% THF-TFA
CCF
~-anU N~
To a solution of 4-(2-benzyloxy-6-benzyloxycarbonyl)-3,5-dibenzyloxybenzoic acid (304.7 mg, 0.449 mmol) in CH2Cl2 (3ml) was added 2 drops of DMF and oxalyl chloride (2.0 M
solution in CH2C12, 561 ~L, 1.123 mmol) at room temperature.
The mixture was kept stirring at room temperature for 1 hr.
Solvents were removed and the acid chloride residue was taken into CH2C12 (5ml) after drying under vaccuum for 1 hr.
A solution of the lactam alcohol (200 mg, 0.449 mmol), Et3N (227.19 mg, 312.9 ~L, 2.245 mmol) and DMAP (10.97 mg, 0.089 mmol) in CH2C12 (5ml) was treated with the acid chloride-CH2Cl2 solution made above at 5-C. The reaction mixture was allowed to stir at room temperature for 3 hr and then chromatographed on silica gel eluting with 5:3 to 1:1 hexane:EtOAc. The product was obtained as white solid (250 mg, 51%).
W24f~67 4 - 212 - PCT~S94/02283 COMPO~ND 523 Compound 612 (225 mg, 0.204 mmol) was dissolved in THF
(20 ml) and treated with a few drops of TFA and Pd(OH)2 (70 mg, 30% by weight on carbon). The mixture was subject to hydrogenolysis with a H2 balloon for 5 hr. THF was removed in vacuo and the residue taken into MeOH. The MeOH solution was concentrated after filtering through a pad of celite to give Compound 523 as a yellow solid (126 mg, 95%). m.p. 174-176 (dec)-C; lH NMR (CD30D) ~ 7.90 (d, J = 8.4Hz, lH, NH), 7.42 (d, J = 8.7Hz, 2H, ArH), 7.26 (d, J = 7.7Hz, lH, ArH), 7.14-7.12 (s, br, 5H, ArH), 7.04 (t, J = 8.1Hz and J = 7.7Hz, lH, ArH), 6.78 (d, J = 8.1Hz, lH, ArH), 6.65 (s, 2H, ArH), 6.57 (d, J = 8.6Hz, 2H, ArH), 5.22 (m, lH, H-3). 4.92 (m, lH, H-2), 4.52 (d, J = 14.5Hz, lH, NCHPh), 4.42 (d, J = 14.5Hz, NCHPh), 3.59 (t, br, lH, H-6), 3.22 (dd, br, lH, H-6), 1.87 (m, 2H, H-5), 1.62 (m, lH, H-4), and 1.32 (m, lH, H-4); IR (KBr) cm1 3398, 1717, 1708, 1635, 1606, and 1543. Anal. Calcd. for C2SH30NZOll-l-25H2O C, 61.67; H, 4.88; N, 4.12. Found: C, 61.78;
H, 5.02; N, 3.92. LRFAB (M + 1) : 655.
_ W 0 94/20062 2 132 1 5 7 ~ 1 2 ~- R~-4- t~2,6-Dichlorohen~oyl)-3,5- ~ihy ~. GxyL e~zoyloxy]-3- R~- t~-hy~roxybenzamido)perhydro~zepine trifluoro~cetic ~cid tCOM PO~nND 52 ~) Br BnO~_OBn1)BuLi,THF,-72c Cl O HO~
- ~J l~ C H O ~CO2t-Bu - CO2t-Bu 2) r~ ~ Cl OBn ~CI
Jones Reagent Cl O OBn Cl O OBn ~COOH , HCOOH ~CO2t-Bu Cl OBn Cl OBn 1)(C O C1)2 ,OH
2) (~HN ~OBn ~ Ph PH320~7A
Et3N, DMAP
0~ 0~
O BnO 1)TFA
~HN ~ OBn ' ~H ~OH
- ~NJ O Pd(OH)2 t C ~ J o Ph TFA
W094/2~62 21~ 7 4 ~ ~ PCT~S94/02283 1,1-Dimethylethyl-3,5-~; hen ~yloxy-~-(2,6-~ichloromethylphenyl-hydroxy)benzoate 2.80 ml (7.03 mmole) of 2.5 M n-butyllithium was added dropwise to a cold solution (-72- C) of 3.00 g (6.39 mmole) of 1,1-dimethylethyl-4-bromo-3,5-dibenzyloxybenzoate in 40 ml of anhydrous tetrahydrofuran (THF) under nitrogen. The temperature of the solution was maintained below -70 C
throughout the addition and the solution was stirred in the cold for ten minutes. A solution of 1.20 g (7.03 mmole) of 2,6-dichlorobenzaldehyde in 10 ml of THF was added dropwise to the solution maintaining the temperature below -65 c throughout the addition. The solution was allowed to warm to room temperature as it stirred for two hours. The solution was partitioned between 20 ml of lN hydrochloric acid and 50 ml of ethyl acetate while stirring for five minutes. The organic layer was separated, washed with water, saturated brine and dried over magnesium sulfate. The solvent was evaporated and the residue was recrystallized from ethanol to yield 1.90 g (55%) of white crystals. Anal. Calcd for C30H30Cl205: C, 67.97;
H, 5.35. Found: C, 67.99: H, 5.20 1,1-Dimethylethyl-~-(2,6-dichlorobenzoyl)-3,5-dibenzyloxybenzoate To a cold solution (0- C) of 1,1-dimethylethyl-3,5-dibenzyloxy-4-(2,6-dichlorophenylmethylhydroxy)benzoate (1.00 g, 1.77 mmole) in 20 ml of acetone was added 5 ml of Jones reagent. As the Jones reagent was being added the solution became very thick. Therefore, an additional 20 ml of acetone was added. The reaction mixture was allowed to stir at room temperature for five hours. Isopropyl alcohol was added to the solution to destroy the Jones reagent. The reaction mixt~re was filtered through celite and washed with acetone. The solvent was removed in vacuo. The residue was taken up in 200 ml of ether. The ether solution was washed with saturated sodium bicarbonate, saturated brine and dried over magnesium sulfate. ~he solvent was evaporated to yield 0.79 g (79%) of a white solid.
~094/20062 PCT~S94/02283 4-(2,6-DichlorobQnzoyl)-3,5-~ihQnPyloxybenzoic acid A solution of 0.95 g of the prime compound (1.70 mmole) in 20 ml of formic acid was stirred at room temperature for three hours. The solution was poured over ice water, collected and dried. Recrystallized from toluene to yield 0.75 g (87%) of yellow crystals.
Trans-N-Benzyl-4-t~-~2,6-~ichlorobenzoyl)-3,5-dibenzyloxy-benzoyloYy]-3-(~-benzyloxybenz~mido)perhydroazepine (COMPOUND 608) A solution of 0.40 g (0.79 mmole) of 4-(2,6-dichlorobenzoyl)-3,5-dibenzyloxybenzoic acid in 20 ml of methylene chloride containing a trace (approximately 1 ~L) of dimethylformamide was cooled to 0-C. A 2.0 M solution of oxalyl chloride (0.45 ml, 0.89 mmole) was added and the solution was stirred under a nitrogen atmosphere at room temperature for two hours. The solvent was removed in vacuo.
The residue was taken up twice in 20 ml portions of methylene chloride and the solvent was removed in vacuo. The residue was dissolved in 8 ml of methylene chloride and added dropwise to a solution of 0.38 g (0.89 mmole) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyperhydroazepine, 0.06 ml (0.89 mmole) of triethylamine and 4.00 mg of DMAP in 10 ml of methylene chloride. The solution was stirred at room temperature under nitrogen for twenty-four hours. The solution was diluted with 30 ml of methylene chloride and washed with saturated sodium bicarbonate, saturated brine and dried over magnesium sulfate. The solvent was removed in vacuo The residue was chromatographed on silica gel eluting with hexane - ethyl acetate (70 : 30). Yield 0.30 g (41%) of a clear oil.
4-R*-4-[(2,6-dichlorohenPoyl)-3,5-~ihydl GAYb~ZOY1OAY] - 3 - R* - ( ~ -hy~roxybenz~mido)perhy~roazepine trifluoroacetic aci~
(COMPOUND 524) A solution of 3.00 g (0.33 mmole) of trans-N-benzyl-4-[4-(2,6-dichlorobenzoyl)-3,5-dibenzyloxybenzoyloxy)-3-(4-benzyloxybenzamido)perhydroazepine in 15 ml of ethanol was W094/20062 ~15 7 41~ PCT~S94/02283 treated with 0.05 ml (0.65 mmole) of trifluoroacetic acid. The solution was stirred at room temperature for five minutes. The solvent was removed in vacuo. The residue was treated twice with 15 ml portions of ethanol and evaporated to remove the excess trifluoroacetic acid. The residue was taken up in 15 ml of ethanol, cooled to 0- C, and 0.05 g of moist 10% palladium hydroxide on carbon was added. The mixture was then stirred under an atmosphere of hydrogen for sixteen hours at room temperature. The mixture was filtered, evaporated and the residue was chromatographed on a 41 X 250 mm C 18 column (solvent A: 9S : 5 water/acetonitrile + 0.1% TFA; solvent B:
100% acetonitrile;gradient 0-50% B over 60 min., flow 25 ml /
min.). The pure fractions were pooled to yield 22.7 mg (10%) of a yellow powder, of Compound 524, mp 179 - 183-C. IR (KBr):
3425, 2875, 1677, 1607, 1508, 1429, 1370, 1239, 1200, 777 cm~l;
Anal Calcd for C27H24C12N2O7-H20-1.4 C2HF302: C, 48.56; H, 3.74; N, 3.80. Found: C, 48.57; H, 4.02; N, 3.46.
W094/20062 ~5 7 4 12 PCT~S94102283 Tr~ns-~-R~-t~-(2-tr~ns-c~rboxycyclohexylc~rbonyl~-3,5-~ihydroxybQnzyloxy]-3-R~ -hy~roxybenz~mi~o)perhydro~zepine trifluoro~cetic ~cid s~lt (COMPOUND 525) - HOOC O OBn BnO~3,0Bn 1)BuLi. THF, -72Qc (~CO2t-Bu CO2t-Bu 2) C~
BnBr DMF
BnOOC O OBn BnOOC O OBn [~ , HCOOH ~
BnO CO2H BnO CO2t-Bu 1) (COCI)2 OH
N~OBn o ~Ph ~OEn 1)TFA , O~
~$ HN~OBnPd(OH)2 / C'~S H~_o H
L HTFA
Ph 609 2157~12 W094/2~62 PCT~S94/Ot283 -~ Dimethylethyl-3,5-~iben~yloxy-4-~2-tr~ns-c~ y~lohe carbonyl)benzoate 2.8 ml (7.03 mmole) of 2.5 M n-butyllithium was added dropwise to a cold solution (-72 C) of 3.0 g (6.39 mmole) of 1,1-dimethylethyl-4-bromo-3,5-dibenzyloxybenzoate in 40 ml of anhydrous tetrahydrofuran (THF) under nitrogen. The solution was stirred in the cold for ten minutes. A solution of 1.2 g (7.03 mmole) of trans-1,2-cyclohexanedicarboxylic anhydride in 10 ml of THF was added dropwise maintaining the temperature below -70 C. The solution was stirred in the cold for two hours. The solution was poured into lSO ml of saturated ammonium chloride and 350 ml of ether. The reaction mixture was stirred for thirty minutes. The organic layer was separated, washed with 0.1 N hydrochloric acid, saturated brine and dried over magnesium sulfate. The solvent was evaporated to yield 3.4 g (98 % ) of a clear oil.
1,1-Dimethylethyl-~ -(2-trans-benzyloxycarbonyl-cyclohexylcarbonyl))-3,5-dibenzyloxybenzoate To a solution of 3.40 g (6.20 mmole) of 1,1-dimethylethyl-3,5-dibenzyloxy-4-(2-trans-carboxycyclohexyl-carbonyl)benzoate in 20 ml of dry DMF was added 0.90 g (6.20 mmole) of potassium carbonate and 0.70 ml (6.20 mmole) of benzyl bromide. The solution was stirred at room temperature under nitrogen for two hours. The solution was poured over 100 ml of ice water, extracted twice with 150 ml portions of ether.
The ether solution was washed with water, saturated brine and dried over magnesium sulfate. The solvent was evaporated and the residue was chromatographed on silica gel eluting with 5 %
- 10 % ethyl acetate - hexane to yield 1.0 g (25 % ) of a clear oil.
4-~ 2-trans-benzyloxycarbonylcyclohexylcarbonyl)-3,5-dibenzyloxybenzoic acid A solution of 1.10 g (1.17 mmole) of 1,1-dimethylethyl-4-(4-(2-trans-benzyloxycarbonylcyclohexyl-carbonyl)-3,5-dibenzyloxybenzoate in 10 ml of formic acid was 21~74~2 W094/2~62 PCT~S94/0~283 stirred at room temperature for four hours. The solution was poured over ice water, collected and dried to yield 0.78 g (82.8%) of a white solid. Anal. Calcd for C36H34O7 : C, 73.63 ;
H, 6.18. Found : C, 73.75 ; H,6.08.
Tr~ns-N-benzyl-~ -(2-tr~ns-benzyloxyc~rbonylcyclohexyl-c~rbonyl)-3-~-benzyloxybenz~mi~o)perhydroazepine (CONPO~ND 609) A solution of 0.40 g (0.69 mmole) of 4-(4-(2-trans-benzyloxycarbonylcyclohexylcarbonyl)-3,5-dibenzyloxybenzoic acid in 8 ml of methylene chloride containing a trace (amount approximately 1 ~L) of dimethylformamide was cooled to 0-C. A
2.0 M solution of oxalyl chloride (0.59 ml, 1.18 mmole) was added and the solution was stirred under nitrogen for 2.5 hours. The reaction mixture was evaporated, and the residue was evaporated twice from 15 ml of methylene chloride. The residue was dissolved in 8 ml of methylene chloride and added to a solution of 0.34 g (0.78 mmole) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine, 0.08 ml (0.78 mmole) of triethylamine and 4.0 mg of DMAP in 10 ml of methylene chloride. The solution was stirred at room temperature under nitrogen for sixteen hours. The solution was diluted with 30 ml of methylene chloride and washed with saturated sodium bicarbonate, saturated brine and dried over magnesium sulfate.
The solvent was evaporated and the residue was chromatographed on silica gel eluting with ethyl acetate - hexane (1 : 4 ).
Yield of 0.39 g (57%) of a clear oil.
Tr~ns-~-R~ 2-trans-¢arboxycyclohexylc~rbonyl)-3,5-~ihydroxybenzyloxy)-3-R~-(4-hy~roxybenz~mido)perhydroazepine trifluoroacetic aci~ salt ~COMPOUND 525) A solution of 0.39 g (0.39 mmole) of trans-N-benzyl-4-(4-(2-trans-benzyloxycarbonylcyclohexylcarbonyl))-3-(4-benzyloxy-benzamido)perhydroazepine in a 20 ml solution of methanol, ethanol and methylene chloride (1:2:2) was treated with 0.06 ml (0.77 mmole) of trifluoroacetic acid for five minutes. The solvent was evaporated, and the methanol, W094/20062 ~lS 7 ~ ~ ~ PCT~S94/02283 ethanol, methylene chloride solvent was added twice more and evaporated. The residue was taken up in 15 ml of ethanol, cooled to 0C and 0.05 g of palladium hydroxide on carbon was added. The mixture was then stirred under an atmosphere on hydrogen for six hours at room temperature. The mixture was filtered, evaporated and the residue was chromatographed on a 41 X 250 mm C18 column (solvent A: 95 : 5 water / acetonitrile + 0.1% TFA; solvent B : 100% acetonitrile ; gradient 0 - 50% B
over 60 min., flow 25 ml / min.). The pure fractions were pooled to yield 57 mg (21%) of Compound 525 as a yellow powder, mp 199 - 204 C. IR (KBr) : 1676, 1607, 1508, 1427, 1365, 1276, 1203, 757 cml. Anal. Calcd for C28H3zN20~ H20 1.2 TFA
: C, 52.50 ; H, 5.10 ; N, 4.02. Found : C, 52.83 ; H, 5.45 :
3.97.
NO94/2~62 215 7 41 2 PCT~S94/02283 Trans-~-t4-(2-carboxy-6-hydroxybenzoyl)-3-benzoyloxy-5-hydroxybenzoyloxy]-3-(~-hy~roxybenzamido)perhydroazepine Trifluoroacetic Aci~ Ralt (COMPOUND 528) HO~
(~. N~ p~ in~ (~ N~ OH
H H
A solution of 9.1 mg (0.013 mmol) of (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in 0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016 mmol) of benzoyl chloride. The mixture was stirred for 24 h at room temperature, after which an additional 6.4 ~L of benzoyl chloride was added. After an additional 24 h, 4 ml of benzoyl chloride was added, and the mixture was allowed to stand for a final period of 24 h, after which the reaction was quenched by the addition of 2 ml of methanol. The mixture was evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min). The pure fractions were pooled and evaporated and then lyophilized from water to give 4.8 mg of Compound 528 as a white fluffy solid. FABMS: m/z 655 (M + H).
W094/2~62 21 S ~ ~1 2 PCT~S94/02283 N-Benzoyl-tr~ns-~-[~-(2-carboxy-6-hy~roxybenzoyl)-3,5-dihydroxybenzoyloxy]-3-(~-hydroxybenzamido)perhydroazepine ~COMr~uNv 529) ~ H~ py~ n~ (~. N~
o~3 A solution of 9.1 mg (0.013 mmol) of (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in 0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016 mmol) of benzoyl chloride. The mixture was stirred for 24 h at room temperature, after which an additional 6.4 ~L of benzoyl chloride was added. After an additional 24 h, 4 ~L of benzoyl chloride was added, and the mixture was allowed to stand for a final period of 24 h, after which the reaction was quenched by the addition of 2 ml of methanol. The mixture was evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min). The pure fractions were pooled and evaporated and then lyophilized from water to give 4.6 mg of Compound 529 as a white fluffy solid. FABMS: m/z 677 (M + Na), 655 (M + H).
W094/20062 PCT~S94/02283 - N-Benzoyl-trans-~ 2-~ArhQxy-6-hydlG~yLehzoyl)-3-benzoyloxy-5-hydroxybenzoyloxy]-3-(4-hydroxybenzAmido)perhydroazepine ~COMPOUND 530) (~, N~ OH I~cOC~
o~0 A solution of 9.1 mg (0.013 mmol) of- (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in 0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016 mmol) of benzoyl chloride. The mixture was stirred for 24 h at room temperature, after which an additional 6.4 ~L of benzoyl chloride was added. After an additional 24 h, 4 ~L of benzoyl chloride was added, and the mixture was allowed to stand for a final period of 24 h, after which the reaction was quenched by the addition of 2 ml of methanol. The mixture was evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min). The pure fractions were pooled and evaporated and then lyophilized from water to give 5.3 mg of Compound 530 as a white fluffy solid. FABMS: m/z 781 (M + Na), 759 (M + H).
21~7~1~
W094/2~62 - 224 - PCT~S94/02283 N-Benzoyl-tr~ns-~-t~-(2-~ 6-hy~ L~zoyl)-3-benzoyloxy-5-hydroxybenzoyloxy]-3-(~-benzoyloxybenz~mi~o)perhy~roazepine ~COMPOUND 531) HO~
H~_~ OH PhCOCI ~ H~ O
H O~
A solution of 9.1 mg (0.013 mmol) of (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in 0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016 mmol) of benzoyl chloride. The mixture was stirred for 24 h at room temperature, after which an additional 6.4 ~L of benzoyl chloride was added. After an additional 24 h, 4 ~L of benzoyl chloride was added, and the mixture was allowed to stand for a final period of 24 h, after which the reaction was quenched by the addition of 2 ml of methanol. The mixture was evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min). The pure fractions were pooled and evaporated and then lyophilized from water to give 2.9 mg of Compound 531 as a white fluffy solid. FABMS: m/z 885 (M + Na), 863 (M + H).
W094/2~62 21~ 7 ~12 PCT~S94/02283 Tr~ns-N-benzyl-~-t~-(3-hydroxyphthalido)-3~5-dihydroxybenzoyl-oXy]-3-(~-hyd.Gh~Le~z~;do)perhydroazepine Trifluoroacetic Acid 8alt Hy~rate (COMPOUND 535) ~ OH PhCHO ~
(~, N~ NaCNBH3 ~. N~ OH
H M~OH N
A solution of 15.6 mg (0.022 mmol) of (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in 0.30 ml of methanol was treated with 4.4 ~L (4.6 mg, 0.043 mmol) of benzaldehyde and 21.7 ~1 (0.022 mmol) of a 1 M
solution af sodium cyanoborohydride in tetrahydrofuran. The mixture was stirred for 24 h at room temperature, after which the reaction was quenched by the addition of 0.30 ml of TFA.
The mixture was stirred for 3 h and then evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile ~ 0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min).
The pure fractions were pooled and evaporated and then W094/20062 2 ~ 5 7 ~12 PCT~S94/02283 lyophilized from water to give 10.1 mg of the title compound as a white fluffy solid. FABMS: m/z 625 (M + H). Anal. Calcd for C35H32N2O~ 3 H2O TFA: C, 56.06; H, 4.96; N, 3.53. Found:
C, 55.76; H, 4.87; N, 3.82.
21~74 12 ~+) -anti-3- (~.-c~rboxybenz~mi~o) -~- t3, s-~ihy~roxy-~- ~2, 6-~ihy~roxy) phenylc~rbonyl] benzoyloxyhex~hy~ro~zepine trifluoro~cetic ~ci~ s~lt (COMPO~D 536) OBn ~
, Br~BnO~L OBn ~ DIBAL-H ~qBnOJ~OBu t I OBn BnO~fSL BenzeneBnO~ OBn n-BuU HO~
CN 43%CHO ~ BnO~OBu-48% O
based on 26 % of aldehyde recovery quan. Jones 1. (COC1)2 cat. DMF, ~, CH2C12 BnOJ~ OBn 1~1 2. Et3N, DMAP, ~ BnO~ ~ OBn CH2C12 O ~ OBn KOH-EtOH OBn ~COOBn BnO~ 60% ~1 HO N ~1 ~ o~OH based on 28 % of BnO ~ OBu t 61% ( ~ ester recovery ~ NBn ~-anti BnO ~L OBn ~ OH
~ COOBn H2, Pd(OH)2 s~ N ~ COOH
BnO ~ H ~ lF-TFA HO ~ X~ ~~
~X j2% ~-anti ~,NH CF~OH
~-anV ~_NBn 617 536 The 2,6-dibenzyloxybenzonitrile (4.0 g, 12.68 mmol) was dissolved in benzene (30 ml) and cooled to 5-C. The DIBAL-H (1.0 M in Hexane, 15.2 ml, 15.2 mmol) was then added and the reaction was allowed to warm up to room temperature and stirred for 4 days. H20 (2 ml) was slowly added- to the reaction, followed by 2N HCl until pH was 3Ø Solids precipitated were filtered off and washed with H20 and EtOAc.
WOg4/2~62 215 7 41~ PCT~S94/02283 The organic layer was washed with brine, dried over Na2SO4, and concentrated. The residue was flash chromatographed on silica gel eluting with 6:1 Hexane:EtOAc to yield yellow oil (1.69 g, 43~), which was taken to the next, coupling reaction.
To a solution of t-butyl ester bromide (1.9 g, 4.08 mmol) in THF (40 ml), precooled to -75-C, was added n-BuLi (2.5M, 1.79 ml, 4.49 mmol). The resulting purple solution was allowed to stir at -75C for 30 min, and a solution of aldehyde (1.3 g, 4.08 mmol) in THF (20 ml) was added in a period of 10 min. The mixture was allowed to warm up to -65-C in 20 min.
The color of reaction changed from purple to yellow. The reaction mixture was poured into 0.5N HCl (50 ml) after warming up to 0-C, extracted with EtOAc (100 ml), and washed with brine. The crude material after concentration was purified on a silica gel column eluting with 8:1 Hexane:EtOAc to recover starting material aldehyde (0.5 g, 26~) and yield pure coupling product (1.0 g, 48%, based on 26% of recovered aldehyde).
The coupling product (1.0 g, 1.41 mmol) was dissolved in acetone (40 ml) and treated with Jones's reagent (ca. 2ml) at 5-C until the color of the reaction remained essentially the same color as the Jone's reagent. The reaction was then stirred at room temperature for lhr. Acetone was removed in vacuo and residue was taken into EtOAc, washed with brine, dried over Na2SO4, and concentrated. The pure product was obtained as bright yellow foam from a short silica gel column eluting with 5:1/Hexane:EtOAc (997 mg, 100%).
The t-butyl ester of benzophenone (565mg, 0.779 mmol) was suspended in EtOH-H20 (9:1, 10 ml) and treated with KOH
(2N, 8 ml). The resulting cloudy mixture was warmed to 70-C
for 4 hr. Ethanol was removed in vacuo. The aqueous residue was diluted with EtOAc and water, and acidified by lN HCl. The organic layer was washed with brine, dried over Na2SO4, and concentrated. Flash chromatography of the crude with 5:1 to 1:1 /Hexane:EtOAc afforded recovered starting material ester (160 mg, 28%) and yielded the corresponding acid as a yellow solid (220 mg, 60%, based on 28~ of recovered ester).
W094/2~62 215 7 ~12 PCT~S94/02283 To a solution of benzophenone acid (200 mg, 0.307 mmol) in CH2Cl2 (3ml) was added cat. DMF and oxalyl chloride (2.0 M solution in CH2Cl2, 384 ~L, 0.768 mmol) at room temperature The mixture was stirred at room temperature for 1 hr. Solvents were removed and the acid chloride residue was taken into CH2Cl2 (5ml) after drying over vacuum for lhr.
A solution of azepine alcohol (SPC 104101, 140.8 mg, 0.307 mmol), Et3N (31.1 mg, 43 ~L, 0.307 mmol) and DMAP (7.5 mg, 0.06 mmol) in CH2C12 (5ml) was treated with the freshly made acid chloride-CH2Cl2 solution at 5-C. The reaction mixture was allowed to stir at r. t. for 3 hr and then chromatographed on silic gel eluting with 3:2 / hexane:EtOAc. The product was obtained as pale yellow foam-like solid (205 mg, 61%).
The prior product (200 mg, 0.183 mmol) was dissolved in THF (20ml) and treated with few drops of TFA and 10% Pd(OH)2 (120mg, 62 mol %). The mixture was subjected to hydrogenolysis at 50 psi for 30 hr. THF was removed in vacuo and the residue taken into MeOH. The MeOH solution was concentrated after filtering through a pad of celite and chromatographed on 41 x 300 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1%
TFA: solvent B: 100% acetonitrile; gradient: 0-100% B over 60 min, flow: 25 ml/min). The pure fractions were evaporated to give two yellow solids. The minor product (15.0 mg) remained identified. The major product was found to be Compound 536 (63.5 mg, 52%). m.p. 176-178 (dec)-C; lH nmr (CD30D) ~ 8.06 (d, J = 8.4Hz, 2H, ArH), 7.80 (d, J = 8.5Hz, 2H, ArH) 7.18 (t, J = 8.3 and 8.3 Hz, lH, ArH), 6.94 (s, 2H, ArH), 6.26 (d, J = 8.3Hz, 2H, ArH), 5.42 (m, lH, C~-H), 4.54 (m, lH, C3-H), 3.50 (d, br, 2H, C~-H or C2-H), 2.30 and 2.09 (m and m, lH and 3H, C5-H and C6-H); IR (KBr) cml 3392, 1707, 1676, 1626, and 1593. Anal. Calc. for C28H26N2Ol0-3.0H20-1.0TFA: C, 50.14; H, 4.63; N, 3.90. Found: C, 50.26; H, 4.33; N, 4.21. LRFAB (M +
1) : 551.
W094t20062 215 ~ 412 PCT~S94/02283 anti-~- t ~ - ~ 2 -carboxy- 6 -hydroxybenzoyl ) - 3, 5-dihy~roxybenzoyloxy]-bexahydro-3- ~indole-5-c~rh~y~mido) azepine, trifluoroacetic acid salt ~COMPO~ND 538) anti-hexahy~ro-~.-hydroxy-3- ~in~ole-5-carboxamido)-1-phenylmethylazepine A cooled (5-C) solution of lithium aluminum hydride/tetrahydrofuran (Aldrich, l.ON, 6 ml, 6 mmol) under nitrogen was treated with anti-3-aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.47g, 2.0 mmol) at a rate to keep the pot temperature below lO-C. The mixture was stirred for 16h at room temperature, for 2h at reflux, then cooled on an ice bath. The reaction mixture was treated sequentially dropwise with water (0.23 ml), 15% sodium hydroxide (0.23 ml), and water (0.70 ml), and filtered. The filter cake was rinsed with tetrahydrofuran and the filtrate was concentrated in vacuo to afford the crude perhydroazepine. Meanwhile, a solution of indole-5-carboxylic acid (0.45 g, 2.8 mmol) in anhydrous tetrahydrofuran (8 ml) under nitrogen was treated with 1,1'-carbonyldiimidazole (0.46 g, 2.8 mmol); some bubbling ensued, which subsided after a few minutes. The solution was stirred for 1.5 h, then combined with the crude perhydroazepine. The mixture was stirred at room temperature for 40 h and concentrated in vacuo. The residue was dissolved in methanol (8 ml) and treated with potassium hydroxide (1.0 g) in water (2.0 ml). The solution was stirred at room temperature for 2h, partially concentrated in vacuo, and diluted with water (10 ml). The aqueous suspension was extracted with methylene chloride (3 x 25 ml) containing some 2-propanol, and the combined organic extracts were dried (Na2SO4), concentrated in vacuo, and chromatographed on silica gel (eluted with ethyl acetate) to afford anti-hexahydro-4-hydroxy-3-(indole-5-carboxamido)-l-phenylmethylazepine (0.47 g, 65%) as a white solid.
~0 94/20062 215 7 41~ PCT/US94/02283 anti-~. - t 4 - ~ 2 -carbophenylmethoxy- 6 -phenylmethoxybenzoyl ) - 3,5 -bis- (phenylmethoxy) benzoyloxyl hex~hydro-3- ( indole-5-carbo~c~ido) -1-phenylmethylazepine (COMPO~ID 619) A solution of 4- (2-carbophenylmethoxy-6-phenylmethoxybenzoyl ) -3,5-bis- (phenylmethoxy) benzoic acid (0.24g, 0.35 mmol) in anhydrous methylene chloride (1.2 ml) was treated with N,N-dimethylformamide (2 drops), then with 2.0 N
oxalyl chloride/methylene chloride (0.25 ml , 0.50 mmol), and stirred for one hour under nitrogen. The solution was concentrated in vacuo, placed under high vacuum for one hour, and dissolved in anhydrous methylene chloride (1.5 ml ) . Anti-hexahydro-4-hydroxy-3- ( indole-5-carboxamido) -1-phenylmethylazepine (0.145g, 0.40 mmol) was suspended in anhydrous methylene chloride (1.0 ml), then treated with 4-dimethylaminopyridine ( lOmg), triethylamine (0.10 ml , 0.72 mmol ), and the acid chloride solution prepared above . The mixture was stirred under nitrogen for 17h and concentrated in vacuo . S il ica gel chromatography ( eluted with 3 %
acetone/methylene chloride, then 5% acetone/methylene chloride) af forded anti -4 - t 4 - ( 2 -carbophenylmethoxy-6-phenylmethoxybenzoyl ) -3,5-bis (phenylmethoxy ) benz oyloxy ] hexa-hydro-3-(indole-5-carboxamido)-1-phenylmethylazepine (0.18g, 50%) as a colorless foam.
anti-~- t ~- ~ 2-carboxy- 6-hydroxybenzoyl ) -3, 5-dih~ ~,o..~ Lehzoyloxy] -hexaLy~.o 3- ~ indole-5-c~ rhQY ~mido ) azepine, trifluoroacetic acid salt ~COMPO~JND 538) A solution/suspension of anti-4 - [4 - (2-carbophenylmethoxy-6-phenylmethoxybenzoyl) -3, 5-bis (phenylmethoxy) benzoyloxy ] hexahydro-3 - ( indole-5-carboxamido) -1-phenylmethylazepine (0.13g, 0.127 mmol) in reagent ethanol (5ml) in a 25 ml 2-neck flask under nitrogen was treated with trifluoroacetic acid (30 mg, 0.26 mmol) and with ethyl acetate (0.20 ml, for solubility), then with 20%
Pd(OH)JC (Pearlman's catalyst, 50mg). The flask was fitted with a baloon valve connected to a baloon containing hydrogen, W O 94/20062 215 7 ~1 2 PCTrJS94/02283 then purged with hydrogen and placed under positive hydrogen pressure for 20h. The flask was carefully purged with nitrogen and the solution filtered through celite (wash filter cake with ethanol ), then the f iltrate was concentrated in vacuo . The residue was taken up in methanol ( 15 ml ) and trifluoroacetic acid (0. 5 ml), gravity filtered, and the filtrate was diluted with de-ionized water (75 ml). The mixture was partially concentrated ~ n vacuo, and the aqueous solution was freeze-dried for 18h. Collection of the yellow solid Compound 538, anti-4 - [ 4- ( 2-carboxy-6-hydroxybenzoyl ) -3, 5-dihydroxybenzoyloxy ~ hexahydro-3 - ( indole-5-carboxamido) -azepine, trifluoroacetic acid salt (71mg, 68%); mp 170-180-C. IR (KBr) 1680 , 1635 , 1607 cm; mass spectrum (FAB): m/z 574 ; H N~
(d6-DMS0) ~ 11.67 (s, 2H), 11.37 (s, lH), 9.87 (s, lH), 9.00 -
= 9 Hz), 5.39 (br d, lH, J = 7 Hz), 4.48 (m, lH), 3.00 - 3.20 (m, 4H), 2.05 - 2.20 (m, lH), 1.70 - 2.00 (m, 3H).Anal.
Calcd. for C27H26N20~-2H2O: C, 59.77; H, 5.57; N, 5.16. Found:
C, 59.83; H, 5.39; N, 5.46.
W094l20062 21 S 7 412 PCT~S94tO~83 Anti-4-t3,5-Dihy~roxy-~-(2-hy~roxyphenylc~rbonyl)]benzoyloxy-3-(~-hydroxybenz~mido)azepine ~COMPOUND 506) To a solution of anti-3-(4-benzyloxybenzamido)-4-[3,5-dibenzyloxy-4-(2-benzyloxyphenylcarbonyl)~benzoyloxy-N-benzylazepine (150 mg, 0.156 mmol) in EtOAc/EtOH (6 ml, 1:1) was added Pd(OH)2 (Pearlman's catalyst) (90 mg, 60% on weight basis), and then H2 at atmospheric pressure. After stirring vigorously for 24h at room temperature, the reaction mixture was filtered through a pad of celite. The filtrate was concentrated and purified on flash column (silica gel: 50 ml;
eluted with 20% ethanol in methylene chloride). CompoUnd 506 was obtained as yellow powder (30 mg, 38%): mp 174 - 176-C; lH
NMR (DMSO) ~ 7.65 (d, J = 8.64, 2H, ArH), 7.54 (td, lH, ArH), 7.26 (dd, J = 1.6, 7.9 Hz, lH, ArH), 6.70 (d, J = 6.48, lH, ArH), 6.98 (s, 2H, ArH), 6.87 (td, lH, ArH), 6.77 (d, J = 8.67 Hz, 2H, ArH), 5.18 (m, lH, 4CH), 4.19 (m, lH, 3 CH), 2.94 -2.88 (dd, lH, CH2N), 2.83 - 2.73 (m, 3H, CH2N), 1.91 (m, 2H, 6 CH2), 1.74 and 1.64 (m and m, 2H, 5 CH2); IR (KBr) cm1 3394, 1704, 1623, 1609, and 1504. Anal. calcd. for C2~H26N2O8-1 1/4 H2O: C, 61.30; H, 5.43; N, 5.29. Found: C, 61.33; H, 5.29;
N, 4.96.
W094l2~62 PCT~S94/0~83 21~7~12 - 178 -Tran~ -benzyloxybenz~mido)-2-t~-(2-benzyloxycarbonylbenzo yl)-3,5-dibenzyloxybenzoyloxyl)cyclohept~ne (COMPOUND 507) An oven-dried 25ml 3-neck round bottom flask, under N2~ was charged with trans-2-(4-benzyloxybenzamido) cycloheptanol (204 mg, 0.6 mmol), 4-(2-benzyloxycarbonyl benzoyl)-3,5-dibenzyloxybenzoic acid (376 mg, 0.66 mmol), 1,3-dicyclohexylcarbodiimide (136 mg, 0.66 mmol), 4-dimethyl-aminopyridine (74 mg, 0.66 mmol), and dry methylene chloride (4.5 ml). The resultant suspension was stirred at room temperature for 17 hours, diluted with methylene chloride (6ml), washed with water (5 ml x 3), and dried MgSO4. The solvent was evaporated and the residue was chromatographed (SiO2, 1:1:2/ diethyl ether:methylene chloride:hexane) to give a white powder (429 mg, 80%): mp 153-154-C; ~H NMR (CDCl3) 6.89 - 7.68 (m, 30H), 6.33 (d, J = 8.6 Hz, lH), 5.16 (s, 2H), 5.13 (tm, J = 9.4 Hz, lH), 5.04 (s, 2H), 4.95 (ABq, J = 14.2, 12.2 Hz, 4H), 4.45 (tm, J = 9.3 Hz, lH), 1.55 - 2.08 (m, 10H) IR KBr 3466, 3367, 1735, 1717, 1679 cm 1. Anal. calcd for CS7HS1O~ C, 76.58; H, 5.75; N, 1.57. Found: C, 76.61; H, 5.82; N, 1.35.
21~7~1~
WOg4/20062 - 179 - PCT~S94/02283 Tr~ns~ -(2-Carboxy-6-hy~roxybenzoyl)-3,5-dihydroxy benzoyloxy)-3-(~-hy~roxybenz~mido)~zepineTr~fluoroaceticAci~
8alt (COMPO~ND 508) Trifluoroacetic Aci~ 8alt of Balanol BnO2C~
o~
C02~n BnO~_~ OBn B~ 1~ (COCI)2 ~ OBn r¦ T ~ 2) NEt3, DMAP, r HO2C~OBnOBn ~' N~ OBn ~ H~ OBn HO~
O ~
H2 ~ o =( OH OH
Pd(OH)2 \ OH
HO ~ ~ CF3CO2H
o O HN o (~NH 508 Trans-N-Benzyl~ -(6-benzyloxy-2-(benzyloxyc~rh~nyl)benzoyl)-3,5-~ih6nzyloxybenzoyloxy)-3-(4-benzyloxybenzamido)azepinQ
(COMPOUND 602) A solution of 84 mg (0.12 mmol) of 4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoic acid in 2 ml of methylene chloride containing a trace (approximately 0.5 ~L) of dimethylformamide was cooled to 0-C. Oxalyl chloride (11.9 ~L, 0.136 mmol) was added, and the mixture was stirred under a nitrogen atmosphere for 1.5 h. An additional 11.9 ~L
of oxalyl chloride was added, and the mixture was stirred for an additional 1.5 h. The reaction mixture was evaporated, and the residue was evaporated twice from 15 ml of methylene chloride. The residue was dissolved in 2 ml of methylene chloride, and was added to a solution of 59.1 mg (0.137 mmol) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine, 19.0 ~1 (0.136 mmol) of triethylamine, and 3 mg of DMAP in 1.5 W094l2~62 PCT~S94/02283 2157~12 - 180 _ ml of methylene chloride at 0CC. The mixture was stirred at room temperature under a nitrogen atmosphere for 22 h, after which it was diluted with 30 ml of methylene chloride, washed with saturated sodium bicarbonate and brine, dried over magnesium sulfate, and evaporated to give 139 mg of the crude product. Chromatography on silica gel eluting with 6/4 hexane - ethyl acetate gave 89.4 mg (66%) of Compound 602 as a yellow oil, which was used directly in the next step.
Tr~ns-4-(4-(2-CarhoYy-6-hy~roxybenzoyl)-3,5-di~y ~x~oenzoyloxy) -3-(4-hydroxy~6n~Amido)azepine Trifluoroacetic Acid Salt (COMPO~ND 508) A solution of 65 mg (0.060 mmol) of Trans-N-benzyl-4-(4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoyloxy)-3-(4-benzyloxybenzamido)azepinein30ml of 1/2/2 methanol/ethanol/methylene chloride was treated with 17 ~1 of trifluoroacetic acid and evaporated. The residue was dissolved in 12 ml of 3/1 ethanol/methanol, 15.6 mg of moist 10% palladium hydroxide on carbon was added, and the mixture was ch~ken on a Parr apparatus under 50 psi of hydrogen for 5 h. The mixture was filtered, evaporated, and the residue was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile;
gradient: 0-100% B over 60 min, flow: 15 ml/min). The pure fraction was evaporated and then lyophilized from water to give 11.2 mg (33%) of the title compound as a yellow fluffy solid.
IR (KBr) : 1701, 1674, 1636, 1608, 1425, 1234, 1200 cm-l;
FABMS: m/z 573 (M + Na), 551 (M+H); HRMS: calcd for C28H27N2Olo 551.1665, found 551.1697.
WO 94/20062 - 181 21~ 7 ~12 PCT/US94/02283 ent-B~l~nol (coMpo~ D 509 ) E3n,N~'NH $~CF3 (N~ oCH33 ~%&F3 ~ Cl Bn NH + Bn NH
~OB CH2CI2 ~ d~
OBn OBn 1) SilicaChi~""atography 3% NEt3, 30% EtOAc `~ in hexanes 2) KOH, MeOH
~-OH ~OH
B,N ~ B~N ~
od~ o5~
OBn OBn HO2C~
~OH ~
Bn,N~NH ~HO~_ oOH
o ~ ~d 509 ~ 0~
OBn ~~~. N~ OH
HN
~8)-Mosher's a¢id chloride Note that the chirality label is changed going from acid to acid chloride. (R)- (+)--Methoxy--trifluoromethyl acetic acid (5.1 g, 21.8 mmol) was slurried in hexane (3 ml).
DMF (2 drops) was added followed by a 2M methylene chloride solution of oxalyl chloride (33 ml, 66 mmol). The solution was refluxed for 3 h, cooled to rt, concentrated and distilled by W094/20062 PCT~S94/0~83 21S7~12 - 182 -kugelrohr (T = 45C at 0.1 mm Hg) to give the product as a clear oil.
Trans-N-Benzyl-3-~-benzyloxybenz~mido)-~ R)-2-methoxy-2-trifluoromethylacetoxy)hex~methyleneimine Trans-N-Benzyl-3-(4-benzyloxybenzamido)-4-hydroxy-hexamethyleneimine (1.2 g, 2.79 mmol), DMAP (4 mg, 0.3 mmol) and triethylamine (2.25 g, 3.1 ml, 22.3 mmol) were dissolved in methylene chloride (10 ml) and treated with (S)-Mosher's acid chloride (1.8 g, 1.3 ml, 7 mmol). When TLC indicated complete reaction, the mixture was concentrated and flashed (7 x 15 cm, 3% triethylamine in 4/1 : ethyl acetate/hexanes). The products were separated into clean upper, mixed and clean lower fractions. All three fractions were each again chromatographed on a Dynamax-60 silica column (41.4 mm ID X 30 cm length) using a linear gradient from 20% to 60% B (A = hexanes, B = 10%
triethyl amine in ethyl acetate) over 60 m at 25 ml/min. The clean upper HPLC fractions from the upper and mixed runs were combined (490 mg) for hydrolysis lH-NMR (300 MHz, CDCl3) 1.6-1.8 (2H, m), 1.94 (2H, m), 2.48 (lH, m), 2.77 (lH, m), 2.9-3.0 (2H, m), 3.50 (lH, d, J = 13 Hz), 3.54 (3H, s), 3.72 (lH, d, J = 13 Hz), 4.1-4.2 (lH, m), 5.14 (2H, s), 5.28 (lH, m), 6.84-7.65 (14H, m).
The clean lower HPLC fractions from the lower run were combined (260 mg) for hydrolysis. H-NMR (300 MHz, CDCl3) 1.64-1.8 (2H, m), 1.8-1.94 (2H, m), 2.53 (lH, m), 2.77 (lH, m), 2.9-3.0 (2H, m), 3.50 (lH, d, J = 13 Hz), 3.52 (3H, s), 3.72 (lH, d, J = 13 Hz), 4.1 (lH, m), 5.13 (2H, s), 5.28 (lH, m), 6.84-7.54 (14H, m).
The upper ester fraction (490 mg, 0.76 mmol) was dissolved in methanol (5 ml) and treated with 85% potassium hydroxide (97 mg, 1.52 mmol) dissoved in methanol (5 ml) and stirred for 16 h. The mixture was treated with water (15 ml) and extracted with methylene chloride (2 x 25 ml). The organic layer was concentrated and chromatographed (2.5 x 10 cm, ethyl acetate) to give the chiral alcohol (266 mg) as an oil.
W094/2~62 2 1 5 7 4 1 2 PCT~S94tO~
The lower ester fraction (260 mg, 0.40 mmol) was dissolved in methanol (5 ml) and treated with 85% potassium hydroxide (53 mg, 0.8 mmol) dissoved in methanol (5 ml) and stirred for 48 h. The mixture was treated with water (15 ml) and extracted with methylene chloride (2 x 25 ml). The organic layer was concentrated and chromatographed (2.5 x 10 cm, ethyl acetate) to give the chiral alcohol (154 mg) as an oil.
Balanol benzophenone (255 mg, 375 ~mol) was dissolved in methylene chloride (3 ml) and treated with DMF (3 drops) followed by a 2M methylene chloride solution of oxalyl chloride (244 ~L, 62 mg, 488 ~mol). After stirring for 1 h, the mixture was concentrated and put under vacuum. The residue was dissolved in methylene chloride (5 ml) and added to chiral amidoalcohol (150 mg, 375 ~mol), DMAP (5 mg), triethylamine (157 ~L, 114 mg, 1.13 mmol) in methylene chloride (5 ml).
After stirring for 16 h, the mixture was chromatographed directly (silica gel, 2.5 x 10 cm, 2/3 : ethyl acetate /hexanes) to give the ester (150 mg)as a glass. The glass was dissolved in 1/1 ethanol/methanol (10 ml), treated with trifluoroacetic acid (100 ~L) and Pearlman's catalyst (palladium hydroxide, 15 mg) and stirred under a hydrogen atmosphere (balloon) for 16 h. The catalyst was filtered off and the mixture concentrated. The residue was chromatographed on a Dynamax-60 C18 column (21 X 250 mm) using a linear gradient from 100% A (0.1% TFA and 5~ acetonitrile in water) to 100% B (pure acetonitrile) over 60 m at 15 ml/min. The clean product, which eluted in 24 m., was concentrated to remove acetonitrile and freeze-dried to give a light yellow powder (45 mg, 22%), Compound 509, identical to Balanol by NMR, IR, CHN
and analytical HPLC. Different rotation [~]D25 = +97.8 (c =
0.319 in CH30H).
PCT~S94/02283 ~57~ 184 -~-)-Trans~ -(2-carboxy-6-hy~roxybenzoyl)-3,5-dihy~roxy benzoyloxy)-3-(~-hy~roxybenz~mi~o)azepineTrifluoroaceticAci~
8alt, (-)-Balanol (COMr~uNv 510) 8nO
) (COCI)2 ~ ~ OBn ll l 2) IPr2NEt, DMAP, I
HO2C ~OBnOBn CH ~ OBn ~ H~ OBn Ho2~3 HO~ OH
H2 ~ OH
pd(OH)2 o ~
~NS' HO~ OH
H
Trans-N-Benzyl-~(~-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl) -3,5-~ibenzyloxybenzoyloxy)-3-(~-benzyloxybenz~mido)azepine A solution of 356 mg (0.583 mmol) of 4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoic acid in 10 ml of methylene chloride contAjning a trace (approximately 1 ~L) of dimethylformamide was cooled to O-C. A 2.0 M solution of oxalyl chloride (0.35 ml, 0.70 mmol) was added, and the mixture was stirred under a nitrogen atmosphere for 2 h. An additional 0.35 ml of oxalyl chloride was added, and the mixture was stirred for an additional 1 h. The reaction mixture was evaporated, and the residue was evaporated twice from 20 ml of methylene chloride. The residue was dissolved in 5 ml of methylene chloride, and was added to a solution of 251 mg (0.583 mmol) of trans-N-benzyl-3-(4-benzyloxybenzamido) -4-hydroxyazepine, 122 ~L (0.700 mmol) of diisopropyl-~094/2~62 215 7 412 PCT~S94/02283 ethylamine, and 4.1 mg of DMAP in 9 ml of methylene chloride at 0-C. The mixture was stirred at room temperature under a nitrogen atmosphere for 16 h, after which it was diluted with 75 ml of methylene chloride, washed with saturated sodium bicarbonate and brine, dried over magnesium sulfate, and evaporated to give 690 mg of the crude product. Chromatography on silica gel eluting with 1/1 hexane - ethyl acetate gave 381 mg (60%) of the title compound as a yellow oil. IR (KBr) :
1719, 1655, 1605, 1581, 1456, 1321, 1248, 1111, 744, 697 cm-l.
Anal. Calcd for C~oH62N2Olo-l~5 H2O: C, 75.18; H, 5.86; N, 2.51.
Found: C, 75.16; H, 5.88; N, 2.74.
~-)-Trans~ 2-Carboxy-6-hydroxybenzoyl)-3,5-dihy~roxy benzoylo~y)-3-(~-hydroxybenz~mi~o)azepine tr~fluoroaceticacid ~lt (COHPOUND 510) A solution of 363 mg (0.333 mmol) of trans-N-benzyl-4-(4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-~ihe~yloxybenzoyloxy)-3-(4-benzyloxybenzamido)azepinein30ml of ethanol was treated with 31 ~L of trifluoroacetic acid, cooled to 0-C, and 53.3 mg of moist 10% palladium hydroxide on carbon was added. The mixture was stirred under an atmosphere of H2 for 22 h. The mixture was filtered, evaporated, and the residue was chromatographed on a 41 x 250 mm C18 column (solvent A; 95:5 water/acetonitrile +0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-50% B over 60 min, flow: 25 ml/min).
The pure fractions were pooled and evaporated and then lyophilized from water to give 75.7 mg (32%) of Compound 510 as a yellow fluffy solid, together with an additional 69 mg (29%) of material which was 96% pure by NMR. mp>200 C; [~]25D = -104- (c = 0.111, methanol); IR (KBr) : 1679, 1607, 1509, 1426, 1369, 1241, 1202, 763 cm . Anal. Calcd for C28H26N2O1o 3 H2O -TFA: C, 50.14; H, 4.63, N, 3.90. Found: C, 50.11; H, 4.40; N, 4.01.
W094/2~62 PCT~S94/02283 ~157~12 - 186 -t+) -anti-3- ~ 4-hydroxybenzamido) -~- [ 3, 5-dihy~roxy-~- ~ 2 -hydroxynaphthyl) carbonyl]benzoyloxyazepine trifluoroacetic acid salt ~COMPOIJND 511) To (+)anti-3-(4-benzyloxybenzamido)-4-[3,5-dibenzyloxy -4-(2-benzyloxynaphthyl)carbonyl] benzoyloxy-N-benzylazepine (338 mg, 0.366 mmol) dissolved in absolute ethanol (18 ml) under an atmosphere of nitrogen was added trifluoracetic acid (30 ~1, 0.386 mmol) followed by Pearlman's catalyst (135mg, 40 % by wt). An atmosphere of hydrogen was introduced and the mixture was allowed to stir for 48 h. The catalyst was removed by filtration and the volatiles removed under reduced pressure.
The product was chromatographed on a Dynamax-60 C18 column (41.4 mm ID X 30 cm length) using a linear gradient from 100%
A (0.1% TFA and 5% acetonitrile in water) to 50% B (pure acetonitrile) over 60 m at 25 ml/min. The product elutes in 50 minutes. Removal of the volatiles provided Compound 511 as a yellow solid (99 mg, 38%), mp 165-168-C. IR KBr (disc) cm 3397, 3274, 3121, 2874, 1796, 1776, 1680, 1633, 1606, 1544, 1510, 1461, 1426, 1369, 1344, 1202, 1142, 1109, 1054, 986, 910, 827, 802, 762, 723, 671. Anal. Calcd for C3lH28N20, 2CF3C02H:
C, 54.69; H, 3.93: N, 3.64. Found: C, 54.46; H, 4.06; N, 3.65.
W094l20062 215 7 412 PCT~S94/02283 ~rans-4-(~-~2-Ca Loh~ 6-hy~roxybenzoyl)-3,5-dihy~roxybenzoylo xy)-3-(~-hydroxybenzam~do)-1-(phenylaminocarbonyl)azepine (COMPOUND 512) HO~ HO~
(~. N~_ OH ~~, NH~_ OH
O NH
A solution of 25.2 mg (0.035 mmol) of (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)azepine trifluoroacetic acid salt in 0.30 ml of dry pyridine was treated with 6.5 ~L (7.1 mg, 0.060 mmol) of phenylisocyanate. The mixture was stirred for 3 h at room temperature, after which the reaction was quenched by the addition of 0.5 ml of methanol. The mixture was evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-100% B over 60 min, flow: 15 ml/min).
The pure fractions were pooled and evaporated and then lyophilized from water to give 23.8 mg (95%) of Compound 512 as W094/20062 - 188 - PCT~S94/022~
a yellow fluffy solid, mp >200 C. IR (KBr) : 1705, 1635, 1545, 1506, 1363, 1240, 760 cm . Anal. Calcd for C35H3lN3O11 -2.5 H2O: C, 58.82 H, 5.08; N, 5.88. Found: C, 59.04; H, 4.94;
N, 5.77.
WOg4/20062 21~ 7 412 PCT~S94tO2283 (+)-~nti-3-~-hy~roxybenzami~o)-~-t3,5-~ihy~roxy-~-(2,3,5,6-tetramethylphenyl)carbonyl]benzoyloxyazep~ne trifluoro~cetic ~cid salt (CONPOUND 513) To(+)-trans-3-(4-benzyloxybenzamido)-4-[3,5-~ihen7yl-oxy-4-(2,3,5,6-tetramethylphenyl)carbonyl]benzoyloxy-N-benzyl-azepine (330 mg, 0.364 mmol) dissolved in 1:1 absolute ethanol:ethyl acetate (100 ml) under an atmosphere of nitrogen was added trifluoracetic acid (42 ~1, 0.546 mmol) followed by Pearlman's catalyst (66 mg, 20 % by wt). An atmosphere of hydrogen was introduced and the mixture was allowed to stir for 24 h. The catalyst was removed by filtration and the volatiles were removed under reduced pressure. The product was chromatographed on a Dynamax~-60 C18 column (41.4 mm ID X 30 cm length) using a linear gradient from 100% A (0.1% TFA and 5%
acetonitrile in water) to 50% B (pure acetonitrile) over 60 m at 25 ml/min. The product elutes in 58 minutes. Removal of the volatiles under reduced pressure provided Compound 513 as a yellow solid (185 mg, 75%), mp 157-160-C. IR KBr (disc) cm 3084, 1717, 1682, 1637, 1609, 1559, 1542, 1509, 1474, 1457, 1425, 1375, 1339, 1270, 1234, 1199, 1140, 1108, 1076, 1007, 938, 848, 813, 767, 723, 669. Anal. Calcd for C3lH28N207 -CF3COzH H20: C, 58.40; H, 5.50; N, 4.13. Found: C, 58.26;
H, 5.28; N, 4.03.
wos4l2~K2 PCT~S94/02283 7 157 412 - lgo -tl)-Anti-4-t3~5-~imethoxy-4-~2~6-~ihydroxyphenylcarbonyl)]-3 (4-hydro~ybenz~mi~o)perhydroazepine (COMPOUND 514) H3CO~¢~OCH3 BuU & o~CH~`C02+ o~ OCH~ ~+ Ht~OOH
CO2+
&OBn O~CH CO~H ~ (~N~ o ~d OBn H2 ~ C~O
OH OCH3 ~- N~ OH
H
The above compounds were synthesized using reactions indicated. Compound 514 was isolated as a yellow powder (37 mg, 5.2% overall yield): mp 165-175 C; NMR (D6 DMSO); IR (KBr) cm : 3431 (OH); 1710 (ester); 1625 (ketone). Anal. calc. for C2~H3oN2~-l-5 H2O: C, 60.31; H, 5.76; N, 4.85. Found: C, 60.53;
H, 5.85; N, 4.91. Intermediate Compound 603 was also isolated.
_ W094/2~62 21~ 7 412 PCT~S9410~
syn-4- ( ~- ( 2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyl-amino)hexahy~ro-3-(4-hydroxybenzoylamino)azepine, trifluoro-acetic aci~ salt (COMPOUND 516) Hexahydro-3-(~-phenylmethoxy)benzoyl~mino-1-phenylmethyl-~zepin ~-one ~ A 25 ml 3-neck round bottom flask under nitrogen was charged with 2.0 N oxalyl chloride/methylene chloride (Aldrich, 1.125 ml, 2.25 mmol), diluted with anhydrous methylene chloride (2 ml), cooled (-65-C), and treated dropwise with anhydrous dimethylsulfoxide (0.35 g, 4.5 mmol) in anhydrous methylene chloride (1.2 ml) at a rate to keep the pot temperature below -60-C. The mixture was stirred at -65 + 5C for 30 min, then treated dropwise with a solution of syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.645 g, 1.5 mmol) in anhydrous methylene chloride (1.5 ml) at a rate to keep the pot temperature below -55-C. The mixture was stirred at 55+5-C for 2 h, then treated dropwise with triethylamine (1.5 ml), warmed to room temperature over one hour, and diluted with methylene chloride (10 ml). The organic solution was washed with water (10 ml), saturated aqueous sodium bicarbonate (10 ml), dried (Na2SO4), and concentrated in vacuo. The residue was chromatographed on silica gel (eluted with 5% acetone/
methylene chloride) to afford hexahydro-3-(4-phenyl-methoxy) benzoylamino-l-phenylmethylazepin-4-one (0.53 g, 82%) as a viscous colorless oil.
~exahydro-3-(~-phenylmethoxy)benzoylamino-1-phenylmethyl-azepin-~-one, oxime A solution of hexahydro-3-(4-phenylmethoxy)-benzoylamino-l-phenylmethyl-azepin-4-one (0.87 g, 2.03 mmol) in ethanol (12 ml) was treated with hydroxylamine hydrochloride (0.19 g, 2.73 mmol), followed by 25% methanolic sodium methoxide (Aldrich, 0.20 g, 0.93 mmol), and was heated to 50C
for one hour. The mixture was cooled to room temperature and treated with additional 25~ methanolic sodium methoxide (0.42 g, 1.94 mmol), then concentrated in vacuo to afford hexahydro-WOg4/2~62 PCT~S94/022~
21S7~12 - 192 -3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepin-4-one oxime (0.89 g, 99%) as a colorless foam.
syn-4-(3,5-Bis~phenylmethoxy)-4-~2-ca~hophenylmethoxy-6-phenyl-methoxybenzoyl)benzoylaminohexahydro-3-~4-phenylmethoxy)-benzoyl~mino-1-phenylmethylazepine ~COMPOUND 615) A solution of hexahydro-3-(4-phenylmethoxy)-benzoyl amino-l-phenylmethyl-azepin-4-one oxime (0.40 g, 0.90 mmol) in reagent methanol (25 ml) in a Parr bottle was treated with Raney Nickel (Aldrich, quarter tsp.), then subjected to hydrogenation at 49-50 psi for six hours. The solution was carefully evacuated of hydrogen, filtered through celite, and the filtrate was concentrated in vacuo to afford 4-aminohexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenyl methylazepine, 1:1 mixture of isomers, which was kept-under nitrogen. Meanwhile, 2'-carbobenzyloxy-2,6,6'-tribenzyloxy benzophenone-4-carboxylic acid (SPC-104034, b.37 g, 0.S5 mmol) was placed in a round-bottom flask and repeatedly covered with toluene and concentrated in vacuo to remove all water and other persistent solvents. Finally, the residue was dissolved in anhydrous methylene chloride (2 ml) under nitrogen, treated with dimethylformamide (3 drops), then with 2.0 N oxalyl chloride/methylene chloride (0.4 ml, 0.8 mmol), and stirred at room temperature for one hour. The solution was concentrated in vacuo, placed under high vacuum for one hour, then dissolved in methylene chloride (3 ml) and added to the 4-aminohexahydro-3-(4-phenylmethoxy)-benzoylamino-1-phenylmethylazepine prepared above. Sodium hydroxide (1.0 N, 1.5 ml) was added, and the mixture was stirred for one hour and separated. The aqueous layer was extracted with methylene chloride (2x10 ml), and the combined organic layer and extracts were washed with saturated sodium chloride (10 ml), dried (Na2SO~), and concentrated in vacuo. The residue was chromatographed (flash) on silica gel (eluted successively with 3% acetone/methylene chloride, 5%
acetone/methylene chloride, and 8% acetone/methylene chloride) to afford, initially, syn-4- ( 3,s-bis(phenylmethoxy)-4- ( 2-carbophenylmethoxy-6-phenylmethoxybenzoyl)-benzoylamino _ V094/2~ 2 1 ~ 7 4 1 2 PCT~S94/0~
hexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenylmethyl-azepine (0.26 g, 43%), then anti-4-(3,5-bis(phenylmethoxy)-4-(2-carbophenyl-methoxy-6-phenylmethoxybenzoyl)benzoylamino hexahydro-3-(4-phenyl-methoxy)benzoylamino-1-phenylmethyl azepine (0.21 g, 35%) as colorless foams. The combined yield was 0.47 g (78%).
syn~ - (2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyl-~m~no)hexahydro-3-(~-hy~roxybenzoyl~m~no)azepine, trifluoro-acetic acid salt (CO~PO~ND 516) A solution of anti-4-(3,5-bis(phenylmethoxy)-4-(2-carbophenylmethoxy-6-phenylmethoxybenzoyl)benzoylaminohexahydro -3-(4-phenylmethoxy)-benzoylamino-1-phenylmethylazepine (0.20 g, 0.183 mmQl) in reagent ethanol (9 ml) and ethyl acetate (1 ml) in a 2-neck 25-ml round bottom flask under nitrogen was treated with Pearlman's catalyst (20% Pd(OH)2/C, 50 mg) and trifluoroacetic acid (42 mg, 0.37 mmol). The flask was fitted with a balloon and a balloon valve, purged with hydrogen, and placed under positive hydrogen pressure for 18 h, then evacuated of hydrogen and purged for several minutes with nitrogen. The solution was carefully filtered through celite (wash filter pad with ethanol) and the filtrate was concentrated in vacuo to a yellow foam. This was dissolved in methanol (20 ml), diluted with deionized water (60 ml), and concentrated in vacuo to remove the methanol. Freeze drying (withoutchromatography)affordedsyn-4-(4-(2-carboxy-6-hydroxy benzoyl)-3,5-dihydroxybenzoylamino)hexahydro-3-(4-hydroxy benzoylamino)azepine (94 mg, 68%) as a voluminous yellow solid;
mp >300-C(dec). Rf (4% acetic acid/ethanol) 0.50; IR (KBr) 1683, 1636, 1604 cm1; 1H NMR (d6-DMS0) ~ 11.66 (s, 2H), 10.07 (s, lH), 9.86 (s, lH), 8.60 - 9.00 (br s, 2H), 8.33 (d, lH, J
= 8 Hz), 8.00 (d, lH, J = 7 Hz), 7.70 (d, 2H, J = 9 Hz), 7.37 (d, lH, J = 8 Hz), 7.27 (t, lH, J = 8 Hz), 7.06 (d, lH, J = 8 Hz), 6.82 (d, 2H, J = 9 Hz), 6.68 (s, 2H), 4.55 (m, lH), 4.40 (m, lH), 3.10 - 3.50 (m, 4H), 1.70 - 2.10 (m, 4H). Anal.
Calcd. for C2~H27N30~-l.5(C2H02F3)-2.0(H20): C, 49.21; H, 4.33; N, 5.55. Found: C, 48.82; H, 4.59; N, 5.79.
2 1 5 ~ 4 1 ~ - 194 - PCT/US94/02283 syn- ~ - t ~ - ( 2 -hydroxycarbonyl- 6-hy~roxybenzoyl ) - 3, 5-0 ihydroxy benzoyloxy] -3- (~-hy~roxybenzamido) perhydroazepine trifluoroacetic aci~ salt (CONPO~rND 517 ) BnO2C O OBn BnO2C O OBn OH OBn O~ COCI ~ H OBn ~,H~ Et3N, DMAP. CH2C12 (~Nb~
Bn Bn 1. Pd(OH)2-C H ~ ~OH 610 EtOH. EtOAc OH OH ~, O
COMPO~lND 610 To a chilled solution (0-5 C) of 4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoic acid (0.52 mmol, 350 mg) in 2 ml of methylene chloride under nitrogen atmosphere was added oxalyl chloride (0.75 mmol, 95 mg) in one portion. N,N-Dimethylformamide (1 drop) was added and the light brown solution was stirred at 0 C for 1.5 h. The solvent and excess oxalyl chloride were then removed in vacuo, the resulting brown oil was redissolved in methylene chloride (2 ml) and added to a cooled solution (0-5 C) of syn-N-benzyloxycarbonyl -3 - ( 4 -hydroxybenzamido ) -4 -hydroxyperhydroazepine (0.62 mmol, 226 mg), triethylamine (1.29 mmol, 130 mg) and 4-dimethylaminopyridine (approximately 10 mg) in 2 ml methylene chloride (nitrogen atmosphere). The reaction mixture was allowed to warm to room temperature and ~0 94l20062 2 1 !~ 7 ~ 1 2 PCT/US94/02283 stirred overnight (approximately 18 h) under a nitrogen atmosphere, after which it was diluted with 50 ml of methylene chloride, washed with saturated sodium bicarbonate solution (lo ml), water (10 ml), and brine (10 ml), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give 440 mg of the crude product. Chromatography on silica gel eluting with 4:1-hexane:ethyl acetate gave 399 mg (71%) of the coupled product as a light yellow solid, which was used directly in the next step.
To a solution of syn-N-benzyloxycarbonyl-4-[4-(2-benzyloxycarbonyl-6-benzyloxybenzoyl) -3, 5-dibenzyloxy benzoyloxy]-3-(4-benzyloxybenzamido)perhydroazepine (0.16 mmol, 180 mg) in ethyl acetate (5 ml) and absolute ethanol (10 ml) was added moist 20% palladium hydroxide on carbon (20% w/w, 36 mg). The reaction flask was fitted with a hydrogen balloon and the grey suspension was stirred at room temperature for 18 hours. The reaction flas~c was then purged with nitrogen gas and the solution diluted with chloroform (50 ml), filtered over celite, treated with 1 ml of trifluoroacetic acid, and concentrated in vacuo to give 87 mg of the crude product. The material was purified by HPLC chromatography with a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA;
solvent B: 100% acetonitrile; gradient 0-50% B over 60 minutes, flow: 15 ml/min). The purified fractions were concentrated and lyophilized from water to give 67 mg (63%) of the title compound as a yellow fluffy solid. IR (KBr): 1704, 1688, 1677, 1632, 1608, 1427, 1235, 1201 cm 1; EA (calculated for C28H26N2Ol0-1.2 C2HF3O2-2.0 H2O): C, 50.48; H, 4.35 N, 3.87.
Found: C, 50.51; H, 4.46; N, 3.88.
W094/2 62 - 196 - PCT~S94/0~
I+)-anti-3-t4-hydroxybenzamido)-4-t3,5-dimethoxy-~-(2,6-dimethoxy)benzoyl]benzoyloxylperhydroazepine trifluoroAcetic aci~ salt (COMPOUND 518) To (+)-anti-3-(4-benzyloxybenzamido)-4-t3,5-dimethoxy-4-(2,6-dimethoxy)benzoyl]benzoyl~oxy-N-benzylperhydroazepine (221 mg, 0.291 mmol) dissolved in ethyl acetate (50 ml) under an atmosphere of nitrogen was added trifluoracetic acid (35 ~L, 0.437 mmol) followed by Pearlman's catalyst (44 mg, 20 % by wt on carbon). An atmosphere of hydrogen was introduced and the mixture allowed to stir for 48 h. The catalyst was removed by filtration and the volatiles were removed under reduced pressure. The product was chromatographed on a Dynamax~-60 C18 column (41.4 mm ID X 30 cm length) using a linear gradient from 100% A (0.1% TFA and 5% acetonitrile in water) to 100% B (pure acetonitrile) over 60 m at 25 ml/min. The product elutes in 58 minutes. Removal of the volatiles under reduced pressure provided Compound 518 as a white solid (62 mg, 26%), mp 134-137-C. IR KBr (disc) cml 3430, 3105, 3016, 2948, 2843, 1772, 1677, 1592, 1544, 1508, 1474, 1436, 1411, 1331, 1235, 1203, 1182, 1127, 1111, 950, 917, 848, 798, 766, 720, 620, 602.
Anal. Calcd for C31H34N2Og 2CF3CO2H: C, 52.11; H, 4.50; N, 3.47. Found: C, 52.30; H, 4.50; N, 3.47.
(+)-~nti-3-(~-benzyloxybenzami~o)-~-t3,5-dimethoxy-~-(2,6-dimethoxy)benzoyl]benzoyloxy-N-benzylperhydroazepine ~COMr~uN~ 60~) To a solution of 3,5-dimethoxy-4-(2,6-dimethoxybenzoyl)benzoic acid (221 mg, 0.639 mmol) in anhydrous dichloromethane (10 ml) under an atmosphere of nitrogen at 0-C
was added oxalyl chloride (436 ~L, 2 M in dichloromethane, 0.872 mmol) dropwise over 5 minutes followed by anhydrous dimethylformamide (3 drops). The ice bath was removed and the suspension rapidly turned into a clear yellow solution. The reaction mixture was allowed to stir for 0.5 h at room temperature. The volatiles were removed under reduced pressure and the remaining solid was dried under vacuum for 2.5 h.
_ ~094/2~K2 2 ~ 5 7 412 pcT~ss4lo22~
To a solution of (+)-anti-3-(4-benzyloxybenzamido)-4-hydroxy-N-benzylperhydroazepine (250 mg, 0.581 mmol), triethylamine (267 ~L, 1.92 mmol), and dimethylaminopyridine (7.1 mg, 0.0581 mmol) in anhydrous dichloromethane (10 ml) under an atmosphere of nitrogen at 0C was added a solution of the above generated acid chloride in anhydrous dichloromethane (10 ml) dropwise over 0.5 h. After allowing to stir while warming to room temperature overnight the reaction mixture was diluted with dichloromethane (200 ml) and washed with water (75 ml). The dichloromethane layer was dried over magnesium sulfate, filtered, and the volatiles were removed under reduced pressure to give a crude white solid. The solid was purified using flash column chromatography (silica gel, 9 : 1 dichloromethane and ethyl acetate) to provide Compound 604 as a white solid (266 mg, 60%), mp 82-85-C. IR KBr (disc) cm 3481, 3029, 2939, 2837, 1711, 1676, 1644, 1591, 1530, 1498, 1474, 1407, 1327, 1248, 1177, 1111, 1024, 995, 913, 845, 795, 744, 700, 605. Anal. Calcd for C45H46N20~: C, 71.22; H, 6.11;
N, 3.69. Found: C, 71.08; H, 6.15; N, 3.64.
3,5-dimethoxy-4-(2,6-dimQthoxy)bQnzoylbenzoiG acid To t-butyl-3,5-dimethoxy-4-(2,6-dimethoxybenzoyl) benzoate (2.26 g, 5.62 mmol) under an atmosphere of nitrogen at O C was added formic acid (25 ml) dropwise. The ice bath was removed and the reaction mixture was allowed to stir at room temperature for 8 h. The volatiles were removed under reduced pressure. Recrystallization of the crude solid with 1:1 ethyl acetate:hexanes provided the title compound as a white solid (1.69 g , 87%), mp 209-212-C. H NMR (DMSO-d6) ~ 7.32 (t, 1 H, J = 8 Hz), 7.16 (s, 2 H), 6.64 (d, 2 H, J = 8 Hz), 3.67 (s, 6 H), 3.61 (s, 6 H); IR KBr (disc) cm1 3432, 3199, 3103, 3016, 2936, 2838, 1731, 1653, 1589, 1475, 1429, 1409, 1313, 1258, 1222, 1185, 1128, 1113, 1028, 944, 897, 869, 801, 773, 713, 690, 617. Anal. Calcd for C1~Hl~O7: C, 62.42; H, 5.24. Found:
C, 62.43; H, 5.30.
t-butyl-3,5-dimethoxy-4-(2,6-dimQthoxybenzoyl)benzoatQ
W094/2~62 PCT~S94/0~
2157 ~ 198 -To a solution of t-butyl-3,5-dimethoxy-4-[(2,6-dimethoxyphenyl)hydroxymethyl]benzoate (3.10 g, 7.70 mmol) in acetone (50 ml) at O C was added Jones reagent dropwise until the reaction mixture retained the orange color of the Jones reagent. The reaction mixture was diluted with dichloromethane (500 ml) and washed with water (150 ml). The dichloromethane layer was dried over anhydrous magnesium sulfate, filtered and the volatiles were removed under reduced pressure. The crude residue was purified using flash column chromatography (silica gel, 1 : 8 ethyl acetate / hexane) to provide the title compound as white solid (2.36 g, 88%), mp 54-57-C. lH NMR
(DMSO-d6) ~ 7.32 (t, 1 H, J = 8.5 Hz), 7.11 (s, 2 H), 6.63 (d, 2 H, J = 8.5 Hz), 3.67 (s, 6 H), 3.61 (s, 6 H) 1.55 (s, 9 H);
IR KBr (disc) cm1 2974, 2839, 1711, 1685, 1590, 1473, 1434, 1406, 1369, 1330, 1292, 1253, 1163, 1126, 1111, 1032, 957, 915, 849, 796, 765, 705, 669, 610. Anal. Calcd for C22H26O7: C, 65.66; H, 6.51. Found: C, 65.67; H, 6.49.
t-butyl-3,5-dimethoxy-~-~(2,6-dimethoxyphenyl)hydroxy-methyl]benzoate Toasolutionoft-butyl-3,5-dimethoxy-4-bromobenzoate (3.50 g, 11.0 mmol) in anhydrous tetrahydrofuran (60 ml) under an atmosphere of nitrogen with an internal temperature of -78-C
(ether/dry ice) was added n-butyllithium (7.58 ml, 1.6 M in hexanes, 12.1 mmol) dropwise at a rate which did not allow the internal temperature to rise above -65-C. To the reaction mixture was added a solution of 2,6-dimethoxybenzaldeyde (1.83 g, 12.1 mmol) in anhydrous tetrahydrofuran (20 ml) dropwise at a rate which did not allow the internal temperature to rise above -65-C. The reaction mixture was allowed to stir while warming to room temperature over 2 h. The reaction mixture was quenched with solid ammonium chloride and the volatiles were removed under reduced pressure. The crude residue was diluted with ethyl acetate (500 ml) and washed with water (200 ml).
The ethyl acetate layer was dried over anhydrous magnesium sulfate, filtered, and the volatiles were removed under reduced pressure. The crude residue was purified by recrystallization W094/2~62 215 ~ 412 PCT~S94/0~
from 1 : 1 ethyl acetate : hexane which provided a white solid of the title compound (3.2 g, 72%), mp 131-133-C. H NMR
(DMSO-d6) ~ 7.14 (t, 1 H, J = 8.5 Hz), 7.08 (s, 2 H), 6.58 (d, 2 H, J = 8.5 Hz), 6.43 (d, 1 H, J = 10.5 Hz), 5.37 (d, 1 H, J
= 10.5 Hz) 3.76 (s, 6 H), 3.70 (s, 6 H) 1.63 (s, 9 H);IR KBr (disc) cml 3541, 3445, 2978, 2843, 1701, 1651, 1588, 1542, 1477, 1455, 1417, 1366, 1328, 1241, 1161, 1119, 1030, 958, 847, 809, 770, 667. Anal. Calcd for Cz2H2~O7: C, 65.33; H, 6.98.
Found: C, 65.19; H, 6.99.
t-butyl-~-bromo-3,5-~imethoxybenzoate To a solution of 4-bromo-3,5-dimethoxybenzoic acid (9.20 g, 35.2 mmol) in anhydrous dimethylformamide (200 ml) under an atmosphere of nitrogen was added N,N-carbonyl diimidazole (6.29 mg, 38.8 mmol). After the reaction mixture was allowed to stir for 1 h at room temperature, DBU (5.80 ml, 38.8 mmol), and t-butanol (9.97 ml, 106 mmol) were added. The reaction mixture was heated at 80-C for 2 h. The reaction mixture was quenched by the slow addition of water (300 ml).
The solid which formed was collected by suction filtration and washed with water (3 X 30 ml) which provided the title compound as a white solid (5.8 g, 52%), mp 119-121-C. lH NMR (DMSO-d6) ~ 7.17 (s, 2 H), 3.89 (s, 6 H), 1.57 (s, 9 H); IR KBr (disc) cm-l 2979, 2936, 2836, 1708, 1590, 1456, 1408, 1366, 1337, 1258, 1231, 1174, 1124, 1033, 960, 8S7, 798, 761, 643. Anal.
Calcd for C13Hl7BrO4: C, 49.23; H, 5.40. Found: C, 49.05; H, S.36.
~-bromo-3,5-dimethoxybenzoic acid To methyl 4-bromo-3,5-dimethoxybenzoate (10 g, 36.4 mmol, Pharmatech International) was added sodium hydroxide (75 ml, 10 N) and methanol (50 ml). The reaction mixture was heated at 70-C for l.S h. The reaction mixture was cooled to Q-C and slowly acidified using HCl (6N). The solid was collected by suction filtration to provide a white solid of the title compound (9.31 g, 98%), mp 22S-228-C. 1H NMR (DMSO-d6) ~ 13.3 (br s, 1 H), 7.23 (s, 2 H), 3.90 (s, 6 H); IR KBr ~ 5 7 4 1~ PCT~S94/022~ _ (disc) cml 3400, 3064, 2969, 2838, 2745, 1689, 1586, 1461, 1407, 1329, 1276, 1230~, 1189, 1127, 1039, 935, 857, 764, 730, 667, 639. Anal. Calcd for CgHgBr04: C, 41.41; H, 3.47. Found:
C, 41.44; H, 3.40.
_ W094/20~2 2 1 ~ 7 412 PCT~S94/02283 ~3R,~R)-[~-(2,6-Dimethoxybenzoyl)-3,s-~ihy~roxybenzoyloxy~-3-(~-hy~roxybenz~mido)perhy~ro~zepine trifluoro~cetic ~cid ~lt (COMPOUND 519) Br BnO~OBn1)gULi,THF,-72C OMe O ~
~Me ~CO2t-Bu CO2t-Bu2) ~ OMe OBn OMe Jones Reagent OMeo OMe o ~COOH , HCOOH ~ CO2t-Bu OMe OBn OMe OBn 1) (cOc1)2 ,OH
~) (;;~Nb~OBn ~ Ph PH320~7A
Et3N, DMAP
0~ 0~
O BnO 1)TFA
~S--HN ,~OBn 2) H2 ~tNb~O H
~NJ O PdoH)2 / C ~NJ O
- Fh TFA
WO94t2~62 215 7 ~12 PCT~S94/0~
l,l-Dimethylethyl 3,5-dibenzyloxy-~-[~2,6-dimethoxyphenyl)-hydro~ymethyl) benzoate 2.5 ml (7.02 mmole) of 2.5 M n-butyllithium in hexane was added dropwise to a cold solution (-72-C) of t-butyl-4-bromo-3,5-dibenzyloxybenzoate (2.69 g, 5.47 mmole) in 40 ml of anhydrous tetrahydrofuran (THF) under nitrogen. The temperature of the solution was maintained below -70-C
throughout the addition and the solution was stirred in the cold for ten minutes. A solution of 1 g (6.02 mmole) of 2,6-dimethoxybenzaldehyde in 15 ml of THF was added dropwise maintaining the temperature below -65-C throughout the addition. The solution was allowed to warm to room temperature as it stirred for two hours. The solution was partitioned between 20 ml of 1 N hydrocholoric acid and 50 ml of ethyl acetate while stirring fifteen minutes. The organic layer was separated, washed with water, saturated brine and dried over magnesium sulfate. The solvent was removed in vacuo. The residue was recrystallized from ethyl acetate-hexane to give 0.83 g (26%) of the title compound as white crystals, mp 152-155-C. Anal. Calcd for C34H36O7: C, 73.36; H, 6.51. Found:
C, 73.04; H, 6.54.
l,1-Dimethylethyl 4-(2,6-dimethoxybenzoyl)-3,5-dibenzoyl oxybenzoate To a cold solution (O-C) of 0.7 g (1.25 mmole) of 1,1-dimethylethyl 3,5-dibenzyloxy-4-[(2,6-dimethoxyphenyl)-hydroxymethyl) benzoatein 15 ml of acetone was added 4 ml of Jones reagent. The solution was stirred in the cold for two hours. To the solution was added 10 ml of isopropyl alcohol to destroy the Jones reagent. The reaction mixture was filtered through celite and washed through with acetone. The filtrate was concentrated in vacuo. The residue was recrystallized from ethanol-water to yield 0.47 g (68%) of the title compound as tan crystals. Anal. Calcd for C34H3407: C, 73.63: H, 6.18.
Found: C, 73.15; H, 6.38.
WO94/2~K2 21~ 7 412 PCT~S94/022~
~-(Z,6-Dimethoxybenzoyl)-3,5-~ibenzyloxybenzoic acid A solution of 0.46 g (0.83 mmole) of l,l-dimethylethyl 4-(2,6-dimethoxybenzoyl)-3,5-dibenzyloxybenzoate in 10 ml of formic acid was stirred at room temperature for two hours.
After stirring for one hour a precipitate formed. The reaction mixture was poured over ice water, and the resultant precipitate was collected and dried to yield 0.36 g (87%) of a tan solid.
Trans-N-Benzyl-4-(~-(2,6-dimethoxybenzoyl)-3,5-~;~^n~yloxy)-3-(4-benzyloxybenz~mido)~zepine (COMPOUND 605) A solution of 0.36 g (0.72 mmole) of 4-(2,6-dimethoxybenzoyl)-3,5-dibenzyloxybenzoic acid in 10 ml of methylene chloride containing a trace (approximately 1 ~L) of dimethylformamide was cooled to 0-C. A 2.0 M solution of oxalyl chloride in methylene chloride (0.41 ml, 0.82 mmole) was added, and the mixture was stirred under a nitrogen atmosphere for ninty minutes. The reaction mixture was evaporated and the residue was evaporated twice from 15 ml of methylene chloride.
The residue was dissolved in 8 ml of methylene chloride and was added to a solution of 0.35 g (0.82 mmole) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine, 0.06 ml (0.82 mmole) triethylamine, and 4.0 mg of DMAP in 10 ml of methylene chloride. The solution was stirred at room temperature under nitrogen for sixteen hours. The solution was diluted with 30 ml of methylene chloride and washed with saturated sodium bicarbonate, saturated brine, dried over magnesium sulfate, and the solvent was removed in vacuo. The residue was chromatographed on silica gel eluting with hexane-ethyl acetate (70:30). Yield 0.33 g (43%) of a glassy oil which solidified on standing.
(3R,~R)-t~-(2,6-Dimethoxybenzoyl)-3,5-dihy~roxybenzoyloxy]-3-(~-hydroxybenzamido)perhydroazQpine trifluoroacetic acid salt (COMPO~ND 519) A solution of 0.33 g (0.36 mmole) of trans-N-benzyl-4-(4-(2,6-dimethoxybenzoyl)-3,5-dibenzoyloxy)-3-(4-benzyloxy W094/20062 - 204 - PCT~S94/02283 benzamido)azepine in 20 ml ethanol-methylene chloride (1:1) was treated with 0.055 ml (0.720 mmole) of trifluoroacetic acid.
The solution was stirred for five minutes. The solvent was evaporated. The ethanol-methylene chloride solvent was added twice more and evacuated in order to remove excess trifluoroacetic acid. The residue was taken up in 15 ml of ethanol, cooled to O-C, and 0.5 g of moist 10% palladium hydroxide on carbon was added. The mixture was then stirred under an atmosphere of hydrogen for six hours at room temperature. The mixture was filtered, evaporated, and the residue was chromatographed on a 41 X 250 mm C 18 column (solvent A: 95 : 5 water / acetonitrile + 0.1% TFA; solvent B:
100% acetonitrile; gradient : 0 - 50% B over 60 min., flow: 25 ml / min.). The pure fractions were pooled and evaporated to yield 0.158 g (60%) of Compound 519, a yellow powder, mp 189-193 C. IR (KBr); 1678, 1605, 1508, 1427, 1370, 1250, 1200, 765 cml. Anal. Calcd for C29H30N2O~ 2H2O B 1-.3 TFA: C, 51.65;
H, 4.84: N, 3.81. Found: C, 51.27; H, 4.68; N, 3.62.
Trans-4- ~4- (2-cis-carboxycyclohexylcarbonyl) -3, 5-~ihydroxybenzoyloxy) -3 - ( 4 -hy~roxybenzamido ) ~zepine trifluoroacetic ~cid salt (COMPO~rND 519) HOOC O OBn BnO~,OBn l)BULi. THF, -72Qc [~C02t Bu CO2t-Bu 2) ~O
BnBr DMF
BnOOC O OBn BnOOC O OBn [J~ , HCOOH ~¢,~
BnO CO2H BnOCO2t-Bu 1) (cOc1)2 ~O H
2) ( )~...N~OBn N O
~Ph ~OBn 1)TFA 0~
(~... N~OBnPd(OH)2 / C (~S~N~O H
- ~ Fh TFA
1, 1-Dimethylethyl 3, 5-dibenzyloxy-4- (2 -Ci9-carboxycyclohexylcarbonyl ) benzoate 2 . 8 ml (7.03 mmole) of 2.5 N n-butyllithium in hexane was added dropwise to a cold solution (-72 C) of 3.0 g (6.39 mmole) of 1,1-dimethylethyl 4-bromo-3,5-dibenzyloxybenzoate in W094/2~62 Z 15 ~ 412 PCT~S94/0~
40 ml of anhydrous tetrahydrofuran (THF) under nitrogen. The solution was stirred in the cold for ten minutes. A solution of 1.2 g (7.03 mmole) of cis-1,2-cyclohexanedicarboxylic anhydride in 10 ml of THF was added dropwise maintaining the temperature below -70 C. The solution was stirred in the cold for two hours. The solution was poured into 150 ml of saturated ammonium chloride and 350 ml of ether. The reaction mixture was stirred for thirty minutes. The organic layer was separated, washed with 0.1 N hydrochloric acid, saturated brine and dried over magnesium sulfate. The solvent was evaporated to yield 3.4 g (98 % ) of a clear oil.
1,1-Dimethylethyl ~ 2-cis-benzyloxycyclohexylcarbonyl))-3,5-dibenzyloxybenzoate To a solution of 3.40 g (6.20 mmole) of 1,1-d im e t h y l e t h y l 3, 5 - d i b e n z y l o xy -4 - (2 - c i s -carboxycyclohexylcarbonyl)benzoate in 20 ml of dry DMF was added 0.43 g (3.10 mmole) of potassium carbonate and 0.33 ml (3.10 mmole) of benzyl bromide. The solution was stirred at room temperature under nitrogen for eight hours. Starting material was still present in the reaction. Therefore, an additional 0.16 ml (1.5 mmole) of benzyl bromide was added and stirring was continued for sixteen hours. The solution was poured over 100 ml of ice water, extracted twice with 150 ml portions of ether. The ether solution was washed with water, saturated brine and dried over magnesium sulfate. The solvent was evaporated and the residue was chromatographed on silica gel eluting with 5 % - 10 % ethyl acetate - hexane to yield 1.0 g (25 %) of a clear oil.
4-(4-~2-cis-benzyloxycyclohQxylc~rbonyl)-3,5-~ih~nzyloxybenzoic ~ci~
A solution of 0.45 g (0.71 mmole) of 1,1-dimethylethyl 4-(4-(2-cis-benzyloxycyclohexylcarbonyl)-3,5-dibenzyloxybenzoate in 10 ml of formic acid was stirred at room temperature for four hours. The solution was poured over ice water, the resultant precipitate was collected and dried to yield 0.34 g (82.8%) of a white solid. Anal. Calcd for C36H34O7 : C, 74.72 ; H, 5.92. Found : C, 74.46 ; H,5.99.
wo g4t20062 2 1 ~ ~ 4 1 2 PCT~S94/0 Trans-N-benzyl-4-(4-~2-cis-bQnzyloxycyclobexylcarbonyl)-3-~4-benzyloxybenz~mido)azepine ~COMPOUND 606) A solution of 0.40 g (0.69 mmole) of 4-(4-(2-cis-benzyloxycyclohexylcarbonyl)-3,5-dibenzyloxybenzoic acid in 8 ml of methylene chloride containing a trace (amount approximately 1 ~L) of dimethylformamide was cooled to o oc.
A 2.0 M solution of oxalyl chloride in methylene chloride (0.59 ml, 1.18 mmole) was added and the solution was stirred under nitrogen for 2.5 hours. The reaction mixture was evaporated, and the residue was evaporated twice from 15 ml of methylene chloride. The residue was dissolved in 8 ml of methylene chloride and added to a solution of 0.34 g (0.78 mmole) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine,0.08 ml (0.78 mmole) of triethylamine and 4.0 mg of DMAP in 10 ml of methylene chloride. The solution was stirred at room temperature under nitrogen for sixteen hours. The solution was diluted with 30 ml of methylene chloride and washed with saturated sodium bicarbonate, saturated brine and dried over magnesium sulfate. The solvent was evaporated and the residue was chromatographed on silica gel eluting with ethyl acetate-hexane (1:4). Yield of 0.5 g (73%) of a clear oil.
Trans-4-~ 2-cis-carboxycyclohexylcarbonyl)-3,5-dihydroxybenzyloxy)-3-(~-hy~roxybenzamido) a zepine trifluoroacetic acid salt (CO~POUND 520) A solution of 0.38 g (0.38 mmole) of trans-N-benzyl-4-(4-(2-cis-benzyloxycyclohexylcarbonyl))-3-(4-benzyloxybenzamido)azepine in a 25 ml mixture of methanol, ethanol and methylene chloride (1 : 2 : 2 ) was treated with 0.06 ml (0.77 mmole) of trifluoroacetic acid for five minutes.
The solvent was evaporated, and the methanol, ethanol, methylene chloride solvent was added twice more and evaporated.
The residue was taken up in 15 ml of ethanol, cooled to 0 C
and 0.05 g of palladium hydroxide on carbon (20% by wt.) was added. The mixture was then stirred under an atmosphere of hydrogen for six hours at room temperature. The mixture was filtered, evaporated and the residue was chromatographed on a WO 94/20062 PCTtUS94tO2283 2157 ~ 208 -41 X 250 mm C18 column (solvent A: 95: 5 water / acetonitrile ~ 0.1% TFA; solvent B : 100% acetonitrile ; gradient O - 50% B
over 60 min., flow 25 ml / min.). The pure fractions were pooled to yield 57 mg (21%) of a yellow powder, mp 122 - 127 C. IR (KBr): 1676, 1607, 1508, 1427, 1365, 1276, 1203, 757 cm . Anal. Calcd for C2ôH32N209 2H20 1.2 TFA : C, 51.18 ; H, 5.25 ; N, 3.92. Found : C, 51.46 ; H, 5.37 ; 3.92.
_ N094/20062 2 15 7 412 PCT~S94/0~
~ Anti-~-t3~5-~ihy~roxy-~-(2~6-~ihy~.ohyLo~zoyl)]hexahy~ro-3 (~-hydroxybenzoyl~mine)~zepine (COMPOUND 521) OBn O OBn BnO ~ OBn OBn OBn OBn O OBn OBn O OBn HCOOH ~J~ (I)BnBr ~
OH OBn C02H (2)KOH OBn OBn CO2H
OH O OH
(I)Courlin~ ~
(2)H~ ~ ~
OH OH O CO ~ OH
,~
) 52 N~ H
tert-butyl 4-(2,6-dibenzyloxybenzoyl)-3,5-dibenzyloxybenzoate (360 mg, 0.51 mmol) was placed in formic acid(10 ml). The resulting suspension was stirred for 20 min and intermittantly heated with a heat gun. The reaction was poured over water (300 ml) and stirred. The solids that precipitated were then filtered. Next, the solids were dissolved in ethyl acetate, and dried over sodium sulfate. The sodium sulfate was filtered off, and the filtrate was concentrated in vacuo and recrystalized in hexane: ethyl acetate to yield a light yellow solid (Acid 175 mg). This ~ ~ 7 ~1 ~ PCT~S94/0~
material was dissolved in dimethylformamide (2 ml). Potassium carbonate (2.2 eq.) was then added. Next, benzyl bromide (5 eq) was added. The reaction was stirred at R.T. for 2 h. The reaction was taken up in ethyl acetate and lN HCl, and placed in a separatory funnel. The organic layer was isolated, dried over sodium sulfate, concentrated in vacuo, and flash chromatographed eluting with hexane:ethyl acetate / 90:10. The major product was isolated as a white foam. This material was then dissolved in a solution of methanol:isopropyl alcohol:water / 45:45:10 with 5% potassium hydroxide. The solution was stirred for 4 h at R.T. The organic solvents were then removed in vacuo, and to the remaining aqueous solution was acidified with lN HCl. Ethyl acetate (100 ml) was then added. The biphasic solution was then placed in a separatory funnel and the organic phase was isolated, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was recrystalized in hexane:ethyl acetate to yield a light yellow solid (92 mg). This benzophenone acid was then converted to Compound 521. Compound 521 was prepared through coupling and reduction in the same was as for Compound 547. A light yellow solid was obtained through reverse phase C-18 HPLC and subsequent lyophilization (7.5 mg). The structure was identified as Compound 521 through 1H NMR.
~094/2~K2 2 1 S 7 ~ 1 2 - 211 - PCT~S94/022~
~ anti-2-(~-~yd~hyL~hzamido)-3-t3,s-dihy~roxy-~-(2-hy~roxy-6-c~rboxyphenylcarbonyl)benzoyloxy]-N-benzylcaprolactam (COMPOUND 523) OBn 1. (COCI)2 cat. DMF, ~OBn B~OOC~L OBn HO N~ 2. Et3N, DMAP, BnO~ OBn =~ OB~ C~'F CH2CI2 ~O N
BnO~ N~_ S1% ~XF
o~OH ~-anti ~ ~-anti ~~ h HOOC ¢~L OH
O I~OH OH H2, Pd(OH)2 HO ~ H _~ 95% THF-TFA
CCF
~-anU N~
To a solution of 4-(2-benzyloxy-6-benzyloxycarbonyl)-3,5-dibenzyloxybenzoic acid (304.7 mg, 0.449 mmol) in CH2Cl2 (3ml) was added 2 drops of DMF and oxalyl chloride (2.0 M
solution in CH2C12, 561 ~L, 1.123 mmol) at room temperature.
The mixture was kept stirring at room temperature for 1 hr.
Solvents were removed and the acid chloride residue was taken into CH2C12 (5ml) after drying under vaccuum for 1 hr.
A solution of the lactam alcohol (200 mg, 0.449 mmol), Et3N (227.19 mg, 312.9 ~L, 2.245 mmol) and DMAP (10.97 mg, 0.089 mmol) in CH2C12 (5ml) was treated with the acid chloride-CH2Cl2 solution made above at 5-C. The reaction mixture was allowed to stir at room temperature for 3 hr and then chromatographed on silica gel eluting with 5:3 to 1:1 hexane:EtOAc. The product was obtained as white solid (250 mg, 51%).
W24f~67 4 - 212 - PCT~S94/02283 COMPO~ND 523 Compound 612 (225 mg, 0.204 mmol) was dissolved in THF
(20 ml) and treated with a few drops of TFA and Pd(OH)2 (70 mg, 30% by weight on carbon). The mixture was subject to hydrogenolysis with a H2 balloon for 5 hr. THF was removed in vacuo and the residue taken into MeOH. The MeOH solution was concentrated after filtering through a pad of celite to give Compound 523 as a yellow solid (126 mg, 95%). m.p. 174-176 (dec)-C; lH NMR (CD30D) ~ 7.90 (d, J = 8.4Hz, lH, NH), 7.42 (d, J = 8.7Hz, 2H, ArH), 7.26 (d, J = 7.7Hz, lH, ArH), 7.14-7.12 (s, br, 5H, ArH), 7.04 (t, J = 8.1Hz and J = 7.7Hz, lH, ArH), 6.78 (d, J = 8.1Hz, lH, ArH), 6.65 (s, 2H, ArH), 6.57 (d, J = 8.6Hz, 2H, ArH), 5.22 (m, lH, H-3). 4.92 (m, lH, H-2), 4.52 (d, J = 14.5Hz, lH, NCHPh), 4.42 (d, J = 14.5Hz, NCHPh), 3.59 (t, br, lH, H-6), 3.22 (dd, br, lH, H-6), 1.87 (m, 2H, H-5), 1.62 (m, lH, H-4), and 1.32 (m, lH, H-4); IR (KBr) cm1 3398, 1717, 1708, 1635, 1606, and 1543. Anal. Calcd. for C2SH30NZOll-l-25H2O C, 61.67; H, 4.88; N, 4.12. Found: C, 61.78;
H, 5.02; N, 3.92. LRFAB (M + 1) : 655.
_ W 0 94/20062 2 132 1 5 7 ~ 1 2 ~- R~-4- t~2,6-Dichlorohen~oyl)-3,5- ~ihy ~. GxyL e~zoyloxy]-3- R~- t~-hy~roxybenzamido)perhydro~zepine trifluoro~cetic ~cid tCOM PO~nND 52 ~) Br BnO~_OBn1)BuLi,THF,-72c Cl O HO~
- ~J l~ C H O ~CO2t-Bu - CO2t-Bu 2) r~ ~ Cl OBn ~CI
Jones Reagent Cl O OBn Cl O OBn ~COOH , HCOOH ~CO2t-Bu Cl OBn Cl OBn 1)(C O C1)2 ,OH
2) (~HN ~OBn ~ Ph PH320~7A
Et3N, DMAP
0~ 0~
O BnO 1)TFA
~HN ~ OBn ' ~H ~OH
- ~NJ O Pd(OH)2 t C ~ J o Ph TFA
W094/2~62 21~ 7 4 ~ ~ PCT~S94/02283 1,1-Dimethylethyl-3,5-~; hen ~yloxy-~-(2,6-~ichloromethylphenyl-hydroxy)benzoate 2.80 ml (7.03 mmole) of 2.5 M n-butyllithium was added dropwise to a cold solution (-72- C) of 3.00 g (6.39 mmole) of 1,1-dimethylethyl-4-bromo-3,5-dibenzyloxybenzoate in 40 ml of anhydrous tetrahydrofuran (THF) under nitrogen. The temperature of the solution was maintained below -70 C
throughout the addition and the solution was stirred in the cold for ten minutes. A solution of 1.20 g (7.03 mmole) of 2,6-dichlorobenzaldehyde in 10 ml of THF was added dropwise to the solution maintaining the temperature below -65 c throughout the addition. The solution was allowed to warm to room temperature as it stirred for two hours. The solution was partitioned between 20 ml of lN hydrochloric acid and 50 ml of ethyl acetate while stirring for five minutes. The organic layer was separated, washed with water, saturated brine and dried over magnesium sulfate. The solvent was evaporated and the residue was recrystallized from ethanol to yield 1.90 g (55%) of white crystals. Anal. Calcd for C30H30Cl205: C, 67.97;
H, 5.35. Found: C, 67.99: H, 5.20 1,1-Dimethylethyl-~-(2,6-dichlorobenzoyl)-3,5-dibenzyloxybenzoate To a cold solution (0- C) of 1,1-dimethylethyl-3,5-dibenzyloxy-4-(2,6-dichlorophenylmethylhydroxy)benzoate (1.00 g, 1.77 mmole) in 20 ml of acetone was added 5 ml of Jones reagent. As the Jones reagent was being added the solution became very thick. Therefore, an additional 20 ml of acetone was added. The reaction mixture was allowed to stir at room temperature for five hours. Isopropyl alcohol was added to the solution to destroy the Jones reagent. The reaction mixt~re was filtered through celite and washed with acetone. The solvent was removed in vacuo. The residue was taken up in 200 ml of ether. The ether solution was washed with saturated sodium bicarbonate, saturated brine and dried over magnesium sulfate. ~he solvent was evaporated to yield 0.79 g (79%) of a white solid.
~094/20062 PCT~S94/02283 4-(2,6-DichlorobQnzoyl)-3,5-~ihQnPyloxybenzoic acid A solution of 0.95 g of the prime compound (1.70 mmole) in 20 ml of formic acid was stirred at room temperature for three hours. The solution was poured over ice water, collected and dried. Recrystallized from toluene to yield 0.75 g (87%) of yellow crystals.
Trans-N-Benzyl-4-t~-~2,6-~ichlorobenzoyl)-3,5-dibenzyloxy-benzoyloYy]-3-(~-benzyloxybenz~mido)perhydroazepine (COMPOUND 608) A solution of 0.40 g (0.79 mmole) of 4-(2,6-dichlorobenzoyl)-3,5-dibenzyloxybenzoic acid in 20 ml of methylene chloride containing a trace (approximately 1 ~L) of dimethylformamide was cooled to 0-C. A 2.0 M solution of oxalyl chloride (0.45 ml, 0.89 mmole) was added and the solution was stirred under a nitrogen atmosphere at room temperature for two hours. The solvent was removed in vacuo.
The residue was taken up twice in 20 ml portions of methylene chloride and the solvent was removed in vacuo. The residue was dissolved in 8 ml of methylene chloride and added dropwise to a solution of 0.38 g (0.89 mmole) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyperhydroazepine, 0.06 ml (0.89 mmole) of triethylamine and 4.00 mg of DMAP in 10 ml of methylene chloride. The solution was stirred at room temperature under nitrogen for twenty-four hours. The solution was diluted with 30 ml of methylene chloride and washed with saturated sodium bicarbonate, saturated brine and dried over magnesium sulfate. The solvent was removed in vacuo The residue was chromatographed on silica gel eluting with hexane - ethyl acetate (70 : 30). Yield 0.30 g (41%) of a clear oil.
4-R*-4-[(2,6-dichlorohenPoyl)-3,5-~ihydl GAYb~ZOY1OAY] - 3 - R* - ( ~ -hy~roxybenz~mido)perhy~roazepine trifluoroacetic aci~
(COMPOUND 524) A solution of 3.00 g (0.33 mmole) of trans-N-benzyl-4-[4-(2,6-dichlorobenzoyl)-3,5-dibenzyloxybenzoyloxy)-3-(4-benzyloxybenzamido)perhydroazepine in 15 ml of ethanol was W094/20062 ~15 7 41~ PCT~S94/02283 treated with 0.05 ml (0.65 mmole) of trifluoroacetic acid. The solution was stirred at room temperature for five minutes. The solvent was removed in vacuo. The residue was treated twice with 15 ml portions of ethanol and evaporated to remove the excess trifluoroacetic acid. The residue was taken up in 15 ml of ethanol, cooled to 0- C, and 0.05 g of moist 10% palladium hydroxide on carbon was added. The mixture was then stirred under an atmosphere of hydrogen for sixteen hours at room temperature. The mixture was filtered, evaporated and the residue was chromatographed on a 41 X 250 mm C 18 column (solvent A: 9S : 5 water/acetonitrile + 0.1% TFA; solvent B:
100% acetonitrile;gradient 0-50% B over 60 min., flow 25 ml /
min.). The pure fractions were pooled to yield 22.7 mg (10%) of a yellow powder, of Compound 524, mp 179 - 183-C. IR (KBr):
3425, 2875, 1677, 1607, 1508, 1429, 1370, 1239, 1200, 777 cm~l;
Anal Calcd for C27H24C12N2O7-H20-1.4 C2HF302: C, 48.56; H, 3.74; N, 3.80. Found: C, 48.57; H, 4.02; N, 3.46.
W094/20062 ~5 7 4 12 PCT~S94102283 Tr~ns-~-R~-t~-(2-tr~ns-c~rboxycyclohexylc~rbonyl~-3,5-~ihydroxybQnzyloxy]-3-R~ -hy~roxybenz~mi~o)perhydro~zepine trifluoro~cetic ~cid s~lt (COMPOUND 525) - HOOC O OBn BnO~3,0Bn 1)BuLi. THF, -72Qc (~CO2t-Bu CO2t-Bu 2) C~
BnBr DMF
BnOOC O OBn BnOOC O OBn [~ , HCOOH ~
BnO CO2H BnO CO2t-Bu 1) (COCI)2 OH
N~OBn o ~Ph ~OEn 1)TFA , O~
~$ HN~OBnPd(OH)2 / C'~S H~_o H
L HTFA
Ph 609 2157~12 W094/2~62 PCT~S94/Ot283 -~ Dimethylethyl-3,5-~iben~yloxy-4-~2-tr~ns-c~ y~lohe carbonyl)benzoate 2.8 ml (7.03 mmole) of 2.5 M n-butyllithium was added dropwise to a cold solution (-72 C) of 3.0 g (6.39 mmole) of 1,1-dimethylethyl-4-bromo-3,5-dibenzyloxybenzoate in 40 ml of anhydrous tetrahydrofuran (THF) under nitrogen. The solution was stirred in the cold for ten minutes. A solution of 1.2 g (7.03 mmole) of trans-1,2-cyclohexanedicarboxylic anhydride in 10 ml of THF was added dropwise maintaining the temperature below -70 C. The solution was stirred in the cold for two hours. The solution was poured into lSO ml of saturated ammonium chloride and 350 ml of ether. The reaction mixture was stirred for thirty minutes. The organic layer was separated, washed with 0.1 N hydrochloric acid, saturated brine and dried over magnesium sulfate. The solvent was evaporated to yield 3.4 g (98 % ) of a clear oil.
1,1-Dimethylethyl-~ -(2-trans-benzyloxycarbonyl-cyclohexylcarbonyl))-3,5-dibenzyloxybenzoate To a solution of 3.40 g (6.20 mmole) of 1,1-dimethylethyl-3,5-dibenzyloxy-4-(2-trans-carboxycyclohexyl-carbonyl)benzoate in 20 ml of dry DMF was added 0.90 g (6.20 mmole) of potassium carbonate and 0.70 ml (6.20 mmole) of benzyl bromide. The solution was stirred at room temperature under nitrogen for two hours. The solution was poured over 100 ml of ice water, extracted twice with 150 ml portions of ether.
The ether solution was washed with water, saturated brine and dried over magnesium sulfate. The solvent was evaporated and the residue was chromatographed on silica gel eluting with 5 %
- 10 % ethyl acetate - hexane to yield 1.0 g (25 % ) of a clear oil.
4-~ 2-trans-benzyloxycarbonylcyclohexylcarbonyl)-3,5-dibenzyloxybenzoic acid A solution of 1.10 g (1.17 mmole) of 1,1-dimethylethyl-4-(4-(2-trans-benzyloxycarbonylcyclohexyl-carbonyl)-3,5-dibenzyloxybenzoate in 10 ml of formic acid was 21~74~2 W094/2~62 PCT~S94/0~283 stirred at room temperature for four hours. The solution was poured over ice water, collected and dried to yield 0.78 g (82.8%) of a white solid. Anal. Calcd for C36H34O7 : C, 73.63 ;
H, 6.18. Found : C, 73.75 ; H,6.08.
Tr~ns-N-benzyl-~ -(2-tr~ns-benzyloxyc~rbonylcyclohexyl-c~rbonyl)-3-~-benzyloxybenz~mi~o)perhydroazepine (CONPO~ND 609) A solution of 0.40 g (0.69 mmole) of 4-(4-(2-trans-benzyloxycarbonylcyclohexylcarbonyl)-3,5-dibenzyloxybenzoic acid in 8 ml of methylene chloride containing a trace (amount approximately 1 ~L) of dimethylformamide was cooled to 0-C. A
2.0 M solution of oxalyl chloride (0.59 ml, 1.18 mmole) was added and the solution was stirred under nitrogen for 2.5 hours. The reaction mixture was evaporated, and the residue was evaporated twice from 15 ml of methylene chloride. The residue was dissolved in 8 ml of methylene chloride and added to a solution of 0.34 g (0.78 mmole) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine, 0.08 ml (0.78 mmole) of triethylamine and 4.0 mg of DMAP in 10 ml of methylene chloride. The solution was stirred at room temperature under nitrogen for sixteen hours. The solution was diluted with 30 ml of methylene chloride and washed with saturated sodium bicarbonate, saturated brine and dried over magnesium sulfate.
The solvent was evaporated and the residue was chromatographed on silica gel eluting with ethyl acetate - hexane (1 : 4 ).
Yield of 0.39 g (57%) of a clear oil.
Tr~ns-~-R~ 2-trans-¢arboxycyclohexylc~rbonyl)-3,5-~ihydroxybenzyloxy)-3-R~-(4-hy~roxybenz~mido)perhydroazepine trifluoroacetic aci~ salt ~COMPOUND 525) A solution of 0.39 g (0.39 mmole) of trans-N-benzyl-4-(4-(2-trans-benzyloxycarbonylcyclohexylcarbonyl))-3-(4-benzyloxy-benzamido)perhydroazepine in a 20 ml solution of methanol, ethanol and methylene chloride (1:2:2) was treated with 0.06 ml (0.77 mmole) of trifluoroacetic acid for five minutes. The solvent was evaporated, and the methanol, W094/20062 ~lS 7 ~ ~ ~ PCT~S94/02283 ethanol, methylene chloride solvent was added twice more and evaporated. The residue was taken up in 15 ml of ethanol, cooled to 0C and 0.05 g of palladium hydroxide on carbon was added. The mixture was then stirred under an atmosphere on hydrogen for six hours at room temperature. The mixture was filtered, evaporated and the residue was chromatographed on a 41 X 250 mm C18 column (solvent A: 95 : 5 water / acetonitrile + 0.1% TFA; solvent B : 100% acetonitrile ; gradient 0 - 50% B
over 60 min., flow 25 ml / min.). The pure fractions were pooled to yield 57 mg (21%) of Compound 525 as a yellow powder, mp 199 - 204 C. IR (KBr) : 1676, 1607, 1508, 1427, 1365, 1276, 1203, 757 cml. Anal. Calcd for C28H3zN20~ H20 1.2 TFA
: C, 52.50 ; H, 5.10 ; N, 4.02. Found : C, 52.83 ; H, 5.45 :
3.97.
NO94/2~62 215 7 41 2 PCT~S94/02283 Trans-~-t4-(2-carboxy-6-hydroxybenzoyl)-3-benzoyloxy-5-hydroxybenzoyloxy]-3-(~-hy~roxybenzamido)perhydroazepine Trifluoroacetic Aci~ Ralt (COMPOUND 528) HO~
(~. N~ p~ in~ (~ N~ OH
H H
A solution of 9.1 mg (0.013 mmol) of (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in 0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016 mmol) of benzoyl chloride. The mixture was stirred for 24 h at room temperature, after which an additional 6.4 ~L of benzoyl chloride was added. After an additional 24 h, 4 ml of benzoyl chloride was added, and the mixture was allowed to stand for a final period of 24 h, after which the reaction was quenched by the addition of 2 ml of methanol. The mixture was evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min). The pure fractions were pooled and evaporated and then lyophilized from water to give 4.8 mg of Compound 528 as a white fluffy solid. FABMS: m/z 655 (M + H).
W094/2~62 21 S ~ ~1 2 PCT~S94/02283 N-Benzoyl-tr~ns-~-[~-(2-carboxy-6-hy~roxybenzoyl)-3,5-dihydroxybenzoyloxy]-3-(~-hydroxybenzamido)perhydroazepine ~COMr~uNv 529) ~ H~ py~ n~ (~. N~
o~3 A solution of 9.1 mg (0.013 mmol) of (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in 0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016 mmol) of benzoyl chloride. The mixture was stirred for 24 h at room temperature, after which an additional 6.4 ~L of benzoyl chloride was added. After an additional 24 h, 4 ~L of benzoyl chloride was added, and the mixture was allowed to stand for a final period of 24 h, after which the reaction was quenched by the addition of 2 ml of methanol. The mixture was evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min). The pure fractions were pooled and evaporated and then lyophilized from water to give 4.6 mg of Compound 529 as a white fluffy solid. FABMS: m/z 677 (M + Na), 655 (M + H).
W094/20062 PCT~S94/02283 - N-Benzoyl-trans-~ 2-~ArhQxy-6-hydlG~yLehzoyl)-3-benzoyloxy-5-hydroxybenzoyloxy]-3-(4-hydroxybenzAmido)perhydroazepine ~COMPOUND 530) (~, N~ OH I~cOC~
o~0 A solution of 9.1 mg (0.013 mmol) of- (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in 0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016 mmol) of benzoyl chloride. The mixture was stirred for 24 h at room temperature, after which an additional 6.4 ~L of benzoyl chloride was added. After an additional 24 h, 4 ~L of benzoyl chloride was added, and the mixture was allowed to stand for a final period of 24 h, after which the reaction was quenched by the addition of 2 ml of methanol. The mixture was evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min). The pure fractions were pooled and evaporated and then lyophilized from water to give 5.3 mg of Compound 530 as a white fluffy solid. FABMS: m/z 781 (M + Na), 759 (M + H).
21~7~1~
W094/2~62 - 224 - PCT~S94/02283 N-Benzoyl-tr~ns-~-t~-(2-~ 6-hy~ L~zoyl)-3-benzoyloxy-5-hydroxybenzoyloxy]-3-(~-benzoyloxybenz~mi~o)perhy~roazepine ~COMPOUND 531) HO~
H~_~ OH PhCOCI ~ H~ O
H O~
A solution of 9.1 mg (0.013 mmol) of (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in 0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016 mmol) of benzoyl chloride. The mixture was stirred for 24 h at room temperature, after which an additional 6.4 ~L of benzoyl chloride was added. After an additional 24 h, 4 ~L of benzoyl chloride was added, and the mixture was allowed to stand for a final period of 24 h, after which the reaction was quenched by the addition of 2 ml of methanol. The mixture was evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min). The pure fractions were pooled and evaporated and then lyophilized from water to give 2.9 mg of Compound 531 as a white fluffy solid. FABMS: m/z 885 (M + Na), 863 (M + H).
W094/2~62 21~ 7 ~12 PCT~S94/02283 Tr~ns-N-benzyl-~-t~-(3-hydroxyphthalido)-3~5-dihydroxybenzoyl-oXy]-3-(~-hyd.Gh~Le~z~;do)perhydroazepine Trifluoroacetic Acid 8alt Hy~rate (COMPOUND 535) ~ OH PhCHO ~
(~, N~ NaCNBH3 ~. N~ OH
H M~OH N
A solution of 15.6 mg (0.022 mmol) of (-)-trans-4-(4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in 0.30 ml of methanol was treated with 4.4 ~L (4.6 mg, 0.043 mmol) of benzaldehyde and 21.7 ~1 (0.022 mmol) of a 1 M
solution af sodium cyanoborohydride in tetrahydrofuran. The mixture was stirred for 24 h at room temperature, after which the reaction was quenched by the addition of 0.30 ml of TFA.
The mixture was stirred for 3 h and then evaporated to a residue which was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile ~ 0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min).
The pure fractions were pooled and evaporated and then W094/20062 2 ~ 5 7 ~12 PCT~S94/02283 lyophilized from water to give 10.1 mg of the title compound as a white fluffy solid. FABMS: m/z 625 (M + H). Anal. Calcd for C35H32N2O~ 3 H2O TFA: C, 56.06; H, 4.96; N, 3.53. Found:
C, 55.76; H, 4.87; N, 3.82.
21~74 12 ~+) -anti-3- (~.-c~rboxybenz~mi~o) -~- t3, s-~ihy~roxy-~- ~2, 6-~ihy~roxy) phenylc~rbonyl] benzoyloxyhex~hy~ro~zepine trifluoro~cetic ~ci~ s~lt (COMPO~D 536) OBn ~
, Br~BnO~L OBn ~ DIBAL-H ~qBnOJ~OBu t I OBn BnO~fSL BenzeneBnO~ OBn n-BuU HO~
CN 43%CHO ~ BnO~OBu-48% O
based on 26 % of aldehyde recovery quan. Jones 1. (COC1)2 cat. DMF, ~, CH2C12 BnOJ~ OBn 1~1 2. Et3N, DMAP, ~ BnO~ ~ OBn CH2C12 O ~ OBn KOH-EtOH OBn ~COOBn BnO~ 60% ~1 HO N ~1 ~ o~OH based on 28 % of BnO ~ OBu t 61% ( ~ ester recovery ~ NBn ~-anti BnO ~L OBn ~ OH
~ COOBn H2, Pd(OH)2 s~ N ~ COOH
BnO ~ H ~ lF-TFA HO ~ X~ ~~
~X j2% ~-anti ~,NH CF~OH
~-anV ~_NBn 617 536 The 2,6-dibenzyloxybenzonitrile (4.0 g, 12.68 mmol) was dissolved in benzene (30 ml) and cooled to 5-C. The DIBAL-H (1.0 M in Hexane, 15.2 ml, 15.2 mmol) was then added and the reaction was allowed to warm up to room temperature and stirred for 4 days. H20 (2 ml) was slowly added- to the reaction, followed by 2N HCl until pH was 3Ø Solids precipitated were filtered off and washed with H20 and EtOAc.
WOg4/2~62 215 7 41~ PCT~S94/02283 The organic layer was washed with brine, dried over Na2SO4, and concentrated. The residue was flash chromatographed on silica gel eluting with 6:1 Hexane:EtOAc to yield yellow oil (1.69 g, 43~), which was taken to the next, coupling reaction.
To a solution of t-butyl ester bromide (1.9 g, 4.08 mmol) in THF (40 ml), precooled to -75-C, was added n-BuLi (2.5M, 1.79 ml, 4.49 mmol). The resulting purple solution was allowed to stir at -75C for 30 min, and a solution of aldehyde (1.3 g, 4.08 mmol) in THF (20 ml) was added in a period of 10 min. The mixture was allowed to warm up to -65-C in 20 min.
The color of reaction changed from purple to yellow. The reaction mixture was poured into 0.5N HCl (50 ml) after warming up to 0-C, extracted with EtOAc (100 ml), and washed with brine. The crude material after concentration was purified on a silica gel column eluting with 8:1 Hexane:EtOAc to recover starting material aldehyde (0.5 g, 26~) and yield pure coupling product (1.0 g, 48%, based on 26% of recovered aldehyde).
The coupling product (1.0 g, 1.41 mmol) was dissolved in acetone (40 ml) and treated with Jones's reagent (ca. 2ml) at 5-C until the color of the reaction remained essentially the same color as the Jone's reagent. The reaction was then stirred at room temperature for lhr. Acetone was removed in vacuo and residue was taken into EtOAc, washed with brine, dried over Na2SO4, and concentrated. The pure product was obtained as bright yellow foam from a short silica gel column eluting with 5:1/Hexane:EtOAc (997 mg, 100%).
The t-butyl ester of benzophenone (565mg, 0.779 mmol) was suspended in EtOH-H20 (9:1, 10 ml) and treated with KOH
(2N, 8 ml). The resulting cloudy mixture was warmed to 70-C
for 4 hr. Ethanol was removed in vacuo. The aqueous residue was diluted with EtOAc and water, and acidified by lN HCl. The organic layer was washed with brine, dried over Na2SO4, and concentrated. Flash chromatography of the crude with 5:1 to 1:1 /Hexane:EtOAc afforded recovered starting material ester (160 mg, 28%) and yielded the corresponding acid as a yellow solid (220 mg, 60%, based on 28~ of recovered ester).
W094/2~62 215 7 ~12 PCT~S94/02283 To a solution of benzophenone acid (200 mg, 0.307 mmol) in CH2Cl2 (3ml) was added cat. DMF and oxalyl chloride (2.0 M solution in CH2Cl2, 384 ~L, 0.768 mmol) at room temperature The mixture was stirred at room temperature for 1 hr. Solvents were removed and the acid chloride residue was taken into CH2Cl2 (5ml) after drying over vacuum for lhr.
A solution of azepine alcohol (SPC 104101, 140.8 mg, 0.307 mmol), Et3N (31.1 mg, 43 ~L, 0.307 mmol) and DMAP (7.5 mg, 0.06 mmol) in CH2C12 (5ml) was treated with the freshly made acid chloride-CH2Cl2 solution at 5-C. The reaction mixture was allowed to stir at r. t. for 3 hr and then chromatographed on silic gel eluting with 3:2 / hexane:EtOAc. The product was obtained as pale yellow foam-like solid (205 mg, 61%).
The prior product (200 mg, 0.183 mmol) was dissolved in THF (20ml) and treated with few drops of TFA and 10% Pd(OH)2 (120mg, 62 mol %). The mixture was subjected to hydrogenolysis at 50 psi for 30 hr. THF was removed in vacuo and the residue taken into MeOH. The MeOH solution was concentrated after filtering through a pad of celite and chromatographed on 41 x 300 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1%
TFA: solvent B: 100% acetonitrile; gradient: 0-100% B over 60 min, flow: 25 ml/min). The pure fractions were evaporated to give two yellow solids. The minor product (15.0 mg) remained identified. The major product was found to be Compound 536 (63.5 mg, 52%). m.p. 176-178 (dec)-C; lH nmr (CD30D) ~ 8.06 (d, J = 8.4Hz, 2H, ArH), 7.80 (d, J = 8.5Hz, 2H, ArH) 7.18 (t, J = 8.3 and 8.3 Hz, lH, ArH), 6.94 (s, 2H, ArH), 6.26 (d, J = 8.3Hz, 2H, ArH), 5.42 (m, lH, C~-H), 4.54 (m, lH, C3-H), 3.50 (d, br, 2H, C~-H or C2-H), 2.30 and 2.09 (m and m, lH and 3H, C5-H and C6-H); IR (KBr) cml 3392, 1707, 1676, 1626, and 1593. Anal. Calc. for C28H26N2Ol0-3.0H20-1.0TFA: C, 50.14; H, 4.63; N, 3.90. Found: C, 50.26; H, 4.33; N, 4.21. LRFAB (M +
1) : 551.
W094t20062 215 ~ 412 PCT~S94/02283 anti-~- t ~ - ~ 2 -carboxy- 6 -hydroxybenzoyl ) - 3, 5-dihy~roxybenzoyloxy]-bexahydro-3- ~indole-5-c~rh~y~mido) azepine, trifluoroacetic acid salt ~COMPO~ND 538) anti-hexahy~ro-~.-hydroxy-3- ~in~ole-5-carboxamido)-1-phenylmethylazepine A cooled (5-C) solution of lithium aluminum hydride/tetrahydrofuran (Aldrich, l.ON, 6 ml, 6 mmol) under nitrogen was treated with anti-3-aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.47g, 2.0 mmol) at a rate to keep the pot temperature below lO-C. The mixture was stirred for 16h at room temperature, for 2h at reflux, then cooled on an ice bath. The reaction mixture was treated sequentially dropwise with water (0.23 ml), 15% sodium hydroxide (0.23 ml), and water (0.70 ml), and filtered. The filter cake was rinsed with tetrahydrofuran and the filtrate was concentrated in vacuo to afford the crude perhydroazepine. Meanwhile, a solution of indole-5-carboxylic acid (0.45 g, 2.8 mmol) in anhydrous tetrahydrofuran (8 ml) under nitrogen was treated with 1,1'-carbonyldiimidazole (0.46 g, 2.8 mmol); some bubbling ensued, which subsided after a few minutes. The solution was stirred for 1.5 h, then combined with the crude perhydroazepine. The mixture was stirred at room temperature for 40 h and concentrated in vacuo. The residue was dissolved in methanol (8 ml) and treated with potassium hydroxide (1.0 g) in water (2.0 ml). The solution was stirred at room temperature for 2h, partially concentrated in vacuo, and diluted with water (10 ml). The aqueous suspension was extracted with methylene chloride (3 x 25 ml) containing some 2-propanol, and the combined organic extracts were dried (Na2SO4), concentrated in vacuo, and chromatographed on silica gel (eluted with ethyl acetate) to afford anti-hexahydro-4-hydroxy-3-(indole-5-carboxamido)-l-phenylmethylazepine (0.47 g, 65%) as a white solid.
~0 94/20062 215 7 41~ PCT/US94/02283 anti-~. - t 4 - ~ 2 -carbophenylmethoxy- 6 -phenylmethoxybenzoyl ) - 3,5 -bis- (phenylmethoxy) benzoyloxyl hex~hydro-3- ( indole-5-carbo~c~ido) -1-phenylmethylazepine (COMPO~ID 619) A solution of 4- (2-carbophenylmethoxy-6-phenylmethoxybenzoyl ) -3,5-bis- (phenylmethoxy) benzoic acid (0.24g, 0.35 mmol) in anhydrous methylene chloride (1.2 ml) was treated with N,N-dimethylformamide (2 drops), then with 2.0 N
oxalyl chloride/methylene chloride (0.25 ml , 0.50 mmol), and stirred for one hour under nitrogen. The solution was concentrated in vacuo, placed under high vacuum for one hour, and dissolved in anhydrous methylene chloride (1.5 ml ) . Anti-hexahydro-4-hydroxy-3- ( indole-5-carboxamido) -1-phenylmethylazepine (0.145g, 0.40 mmol) was suspended in anhydrous methylene chloride (1.0 ml), then treated with 4-dimethylaminopyridine ( lOmg), triethylamine (0.10 ml , 0.72 mmol ), and the acid chloride solution prepared above . The mixture was stirred under nitrogen for 17h and concentrated in vacuo . S il ica gel chromatography ( eluted with 3 %
acetone/methylene chloride, then 5% acetone/methylene chloride) af forded anti -4 - t 4 - ( 2 -carbophenylmethoxy-6-phenylmethoxybenzoyl ) -3,5-bis (phenylmethoxy ) benz oyloxy ] hexa-hydro-3-(indole-5-carboxamido)-1-phenylmethylazepine (0.18g, 50%) as a colorless foam.
anti-~- t ~- ~ 2-carboxy- 6-hydroxybenzoyl ) -3, 5-dih~ ~,o..~ Lehzoyloxy] -hexaLy~.o 3- ~ indole-5-c~ rhQY ~mido ) azepine, trifluoroacetic acid salt ~COMPO~JND 538) A solution/suspension of anti-4 - [4 - (2-carbophenylmethoxy-6-phenylmethoxybenzoyl) -3, 5-bis (phenylmethoxy) benzoyloxy ] hexahydro-3 - ( indole-5-carboxamido) -1-phenylmethylazepine (0.13g, 0.127 mmol) in reagent ethanol (5ml) in a 25 ml 2-neck flask under nitrogen was treated with trifluoroacetic acid (30 mg, 0.26 mmol) and with ethyl acetate (0.20 ml, for solubility), then with 20%
Pd(OH)JC (Pearlman's catalyst, 50mg). The flask was fitted with a baloon valve connected to a baloon containing hydrogen, W O 94/20062 215 7 ~1 2 PCTrJS94/02283 then purged with hydrogen and placed under positive hydrogen pressure for 20h. The flask was carefully purged with nitrogen and the solution filtered through celite (wash filter cake with ethanol ), then the f iltrate was concentrated in vacuo . The residue was taken up in methanol ( 15 ml ) and trifluoroacetic acid (0. 5 ml), gravity filtered, and the filtrate was diluted with de-ionized water (75 ml). The mixture was partially concentrated ~ n vacuo, and the aqueous solution was freeze-dried for 18h. Collection of the yellow solid Compound 538, anti-4 - [ 4- ( 2-carboxy-6-hydroxybenzoyl ) -3, 5-dihydroxybenzoyloxy ~ hexahydro-3 - ( indole-5-carboxamido) -azepine, trifluoroacetic acid salt (71mg, 68%); mp 170-180-C. IR (KBr) 1680 , 1635 , 1607 cm; mass spectrum (FAB): m/z 574 ; H N~
(d6-DMS0) ~ 11.67 (s, 2H), 11.37 (s, lH), 9.87 (s, lH), 9.00 -
9.20 (m, 2H), 8.64 (d, lH, J = 8 Hz), 8.06 (s, lH), 7.56 (d, lH, J = 8 Hz), 7.43 (s, lH), 7.30 - 7.45 (m, 2H), 7.27 (t, lH, J = 8 Hz), 7.05 (d, lH, J = 8 Hz), 6.80 (s, 2H), 6.52 (br s, lH), 5 . 30 (m, lH), 4 . 55 (m, lH), 3 . 25 - 3 . 50 (m, 2H), 3 . 10 -3 . 25 (m, 2H), 2 . 05 - 2 . 20 (m, lH), 1 . 80 - 2 . 05 (m, 3H) . Anal .
Calcd. for C30Hz7N3Og- 1. 9 (C2HO2F3) 2 . 0 (H2O): C, 49 .14; H, 4 . 01; N, 5.09. Found: C, 49.10; H, 4.38; N, 4.88.
NO94/2~62 215 7 412 PCT~S94/02283 anti-~- t ~- ~ 2-~ ~ hY~h ~ 6-hydroxybenzoyl)-3,5-~ihydroxybenzoyl-N-methylamino]hexahy~ro-3-(4-hy~roxybenzoyl~mino)azepine~
trifluoroacetic acid salt ~CONPOUND 539) Ant$-hexahy~ro-3-(4-phenylmethoxy)benzoyl~mino-1-phenylmethyl-~-trlfluoroacetyl~minoazepine ~~ A solution of hexahydro-3-(4-phenylmethoxy)-benzoylamino-l-phenylmethyl-azepin-4-one (0.87 g, 2.03 mmol) in ethanol (12 ml) was treated with hydroxylamine hydrochloride (0.19 g, 2.73 mmol), followed by 25% methanolic sodium methoxide (Aldrich, 0.20 g, 0.93 mmol), and was heated to 50-C
for one hour. The mixture was cooled to room temperature and treated with additional 25% methanolic sodium methoxide (0.42 g, 1.94 mmol), then concentrated in vacuo to afford hexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepin-4-one oxime (0.89 g, 99%) as a colorless foam. A solution of this oxime (1.065 g, 2.4 mmol) in reagent ethanol (45 ml) in a Parr bottle was treated with Raney Nickel (Aldrich, one tsp.) and subjected to hydrogenation in a Parr apparatus at 47 - 51 psi for 5h. The bottle was carefully evacuated of hydrogen and the contents filtered through celite (washed with ethanol under nitrogen). The filtrate was gently concentrated in vacuo at 45-C, diluted with toluene, and further concentrated at -45-C
to remove the remaining ethanol. Meanwhile, a solution of trifluoroacetic acid (0.33 g, 2.9 mmol) in anhydrous tetrahydrofuran (6 ml) under nitrogen was treated with l,l'-carbonyl diimidazole (0.50 g, 3.1 mmol). Some bubbling ensued, and the mixture was stirred for two hours, cooled on an ice bath, and combined under nitrogen with the residual amine prepared above (an additional 2 ml of tetrahydrofuran was used ~ to rinse the CDI adduct into the reaction vessel). The mixture was stirred at room temperature for 18h and concentrated in vacuo, then the residue was chromatographed on silica gel (eluted with 3% acetone/methylene chloride, then with 8%
acetone/methylene chloride) to afford, initially, syn-hexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenylmethyl-4-trifluoroacetylaminoazepine (0.27 g) followed by anti-WO 94/20062 21 S 7 ~ :12 PCT/US94/02283 hexahydro-3-(4-pllenylmethoxy)benzoylamino-1-phenylmethyl-4-trifluoroacetylaminoazepine (0.40 g). The total yield of trifluoroacetamides was 0.67 g (53%); the anti-isomer could be recrystallized from acetonitrile.
~ti-hexahydro-4-~methylamino)-3-(4-phenylmethoxy)benzoyl~ino-l-phenylmethylazep~ne An ice-cooled (5-C) solution of anti-hexahydro-3-(4-phenylmethoxy) benzoyl-amino-1-phenylmethyl-4-trifluoroacetylaminoazepine (0.20 g, 0.38 mmol) in anhydrous N,N-dimethylformamide (2 ml) under nitrogen was treated dropwise with 1.0 N potassium t-butoxide/tetrahydrofuran (Aldrich, 0.40 ml, 0.40 mmol), then stirred for 20 min at room temperature and recooled (5-C). Dimethyl sulfate (38 ~L, 0.40 mmol) was added via syringe, and stirring was continued at 5 C
for 3h. The solution was added to a rapidly stirred mixture of methylene chloride (10 ml) and saturated sodium bicarbonate (5 ml) and the organic layer was separated. The aqueous layer was extracted with methylene chloride (10 ml) and the combined organic solution was dried (Na2S04) and concentrated in vacuo.
The residue was chromatographed on silica gel (eluted successively with 2%, 3%, and 4% acetone/methylene chloride) to afford the methylated intermediate (0.15 g) as a viscous colorless oil. This was dissolved in reagent methanol (1.5 ml), treated with a solution of potassium hydroxide (0.25 g, 4.5 mmol) in water (0.25 ml), and stirred at room temperature~
for 2.5h. The solution was partially concentrated to remove most of the methanol, diluted with water (5 ml), and extracted with methylene chloride (2x15 ml). The combined organic extracts were dried (Na2S04) and concentrated in vacuo to afford anti-hexahydro-4-(methylamino)-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepine (0.105 g, 62%) as a colorless oil.
anti-~- t3,5-Bi~ (phenylmethoxy)-~-~2-carbophQnylmethoxy-6-phenylmethoxybenzoyl)benzoyl-N-methyl~mino]hexahydro-3-(~-phenylmethoxy)benzoyl~mino-l-phenylmethyl~zepine lC-I.r~uNv 620) A solution of 3,5-bis (phenylmethoxy) -4- (2-carbophenylmethoxy-6-phenylmethoxybenzoyl)benzoic acid (0.204 g, 0.30 mmol ) in anhydrous methylene chloride (1.0 ml ) was treated with N,N-dimethylformamide (2 drops), then with 2.0 N
oxalyl chloride/methylene chloride (Aldrich, 0.22 ml, 0.44 mmol ) and stirred for one hour under nitrogen . The solution was concentrated in vacuo, placed under high vacuum for 45 min, then dissolved in methylene chloride (2 ml ) and combined with anti-hexahydro-4- (methylamino) -3- (4-phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.10 g, 0.225 mmol). The mixture was treated with 1.0 N sodium hydroxide (1.0 ml) and stirred for two hours, then diluted with methylene chloride (15 ml ) and water (5 ml). The organic layer was separated and the aqueous solution was extracted with methylene chloride (2x15 ml). The combined organic solution was dried (Na2S0~) and concentrated in vacuo. The residue was chromatographed on silica gel (eluted successively with 5%, then 10% acetone/methylene chloride, then with 3% methanol/methylene chloride) to afford anti-4- [3,5-bis (phenylmethoxy) -4- (2 -carbophenylmethoxy-6-phenylmethoxybenz oyl ) benz oy 1 -N -methy 1 amino ] hexahydro- 3 - (4 -phenylmethoxy)benzoylamino-l-phenylmethyl-azepine (0.22g, 88%) as a viscous colorless oil.
anti-4-t4-~2-c~ 6-hy~.oa~L3nzoyl)-3,5-dihy~roxybenzoyl-N-methyl~mino]hexahydro-3-(~-hydroxybenzoyl~mino)~zepine, trifluoroacetic ~ci~ s~lt (CONPO~ND 539) A solution/suspension of anti-4- [3, 5-bis (phenylmethoxy ) -4 - ( 2 -carbophenylmethoxy-6-phenylmethoxybenzoyl ) benzoyl-N-methylamino ] hexahydro-3 - (4 -phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.22 g, 0.20 mmol ) in reagent ethanol (9 ml ) in a 25 ml 2 -neck round bottom flask was treated with trifluoroacetic acid (50 mg, 0.43 mmol), then with ethyl acetate (1 ml, for solubility) . Pearlman ' s WO 94l20062 21~ 7 412 PCT/US94/02283 catalyst (20% Pd(OH)2/C, so mg) was added, then the flask was quickly purged with nitrogen and fitted with a baloon valve and baloon containing hydrogen. The mixture was purged with hydrogen and kept under positive hydrogen pressure for 20h.
The flask was carefully evacuated of hydrogen and purged for several minutes with nitrogen. The solution was filtered through celite (wash filter cake with ethanol) and the filtrate was concentrated in vacuo and dissolved in N,N-dimethylformamide (1 ml). The solution was loaded onto a 41x250 mm C18 HPLC column and eluted as follows: A-0.1%
TFA/95:5 water:acetonitrile, B-acetonitrile, 100% A to 50:50 A:B over 60 min collected at 25 ml/min. The appropriate fractions were combined and partially concentrated in vacuo, then freeze-dried overnight to afford anti-4-[4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyl-N-methylamino]-hexahydro-3-(4-hydroxybenzoylamino)azepine, trifluoroacetic acid salt (101 mg, 68%) as a voluminous pale yellow solid; mp 285-295-C
(dec). Rf(6:1:1 n-BuOH/AcOH/H20) 0.45; IR (KBr): 1682, 1633, 1620 cm; H NMR (d6-DMSO) ~ 11.68 (br s, 2H), 10.07 (br s, lH), 9.92 + 9.851(s, lH), 8.95 + 8.751 (br s, 2H), 8.28 +
8.181(d, lH, J = 9 Hz), 7.67 + 7.581(d, 2H, J = 9 Hz), 7.20 -7.40 (m, 2H), 7.00 - 7.10 (m, lH), 6.78 - 6.85 (m, 2H), 6.00 (s, 2H), 4.40 - 4.80 (m, 2H), 3.00 - 3.40 (m, 4H), 2.85 +
2.771(s, 3H), 1.80 - 2.10 (m, 4H); mass spectrum(FAB): m/z 564.
Anal. Calcd. for C29H29N3Og-1~1(C2HO2F3)-3~0(H2O) C, 50.43; H, 4.90; N, 5.66. Found: C, 50.32; H, 4.74; N, 5.74.
21~7~12 ~094/2~62 - 237 - PCT~S94/02283 ~-R~-~- [ ( ( (3-hy~roxyc~rbonyl) -2-pyri~inyl) c~rbonyl) -3, 5-Oihy~roxybenzoyloxy] -3-R~ -hy~.o,.yL3nzami~0) perhy~roazepine trifluoroacetic ~ci~ s~lt (COIIrOu~v S~l) BnOOC O OBn BnO
1) (COC1)2 ,~\ ~
BnO CO2HCH2CI2 / DMF . o~oBnN
2) (~---N~OBn ~ NHb~OBn Ph 622 1 ) TFA
2) H2 Pd(OH)2 / C
EtOH
0~
(~,...N~O H
H
TFA
Trans-N-benzyl-~- t ~ ( (3-benzyloxyc~rbonyl) -2-pyridinyl ) c~rbonyl ) -3, 5-dibenzyloxybenzoyloxy] -3- ( ~-benzyloxybenzamido) perhydro~zepine (coMpo~rv 622 ) Carbonyldiimidizole (0.11 g, 0.65 mmole) was added to a solution of 4-[((3-benzyloxycarbonyl)-2-pyridinyl)carbonyl]-3,5-dibenzyloxybenzoic acid ( 0.25 g, 0.44 mmole) in 5 ml of 215 7 '1~ ~ PCT/US94102283 methylene chloride and the solution was stirred at room temperature for sixty minutes under nitrogen. The solution was added to a solution of 0.19 g (0.44 mmole) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyperhydroazepine, 0.12 ml triethylamine, and 5 mg DMAP in 8 ml of methylene chloride.
The solution was stirred at room temperature for twenty hours.
The solution was diluted with 30 ml of methylene chloride, washed with water, saturated brine and dried over magnesium sulfate. The solvent was removed in vacuo. The residue was chromatographed on silica gel eluting with a gradient of 5% -
Calcd. for C30Hz7N3Og- 1. 9 (C2HO2F3) 2 . 0 (H2O): C, 49 .14; H, 4 . 01; N, 5.09. Found: C, 49.10; H, 4.38; N, 4.88.
NO94/2~62 215 7 412 PCT~S94/02283 anti-~- t ~- ~ 2-~ ~ hY~h ~ 6-hydroxybenzoyl)-3,5-~ihydroxybenzoyl-N-methylamino]hexahy~ro-3-(4-hy~roxybenzoyl~mino)azepine~
trifluoroacetic acid salt ~CONPOUND 539) Ant$-hexahy~ro-3-(4-phenylmethoxy)benzoyl~mino-1-phenylmethyl-~-trlfluoroacetyl~minoazepine ~~ A solution of hexahydro-3-(4-phenylmethoxy)-benzoylamino-l-phenylmethyl-azepin-4-one (0.87 g, 2.03 mmol) in ethanol (12 ml) was treated with hydroxylamine hydrochloride (0.19 g, 2.73 mmol), followed by 25% methanolic sodium methoxide (Aldrich, 0.20 g, 0.93 mmol), and was heated to 50-C
for one hour. The mixture was cooled to room temperature and treated with additional 25% methanolic sodium methoxide (0.42 g, 1.94 mmol), then concentrated in vacuo to afford hexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepin-4-one oxime (0.89 g, 99%) as a colorless foam. A solution of this oxime (1.065 g, 2.4 mmol) in reagent ethanol (45 ml) in a Parr bottle was treated with Raney Nickel (Aldrich, one tsp.) and subjected to hydrogenation in a Parr apparatus at 47 - 51 psi for 5h. The bottle was carefully evacuated of hydrogen and the contents filtered through celite (washed with ethanol under nitrogen). The filtrate was gently concentrated in vacuo at 45-C, diluted with toluene, and further concentrated at -45-C
to remove the remaining ethanol. Meanwhile, a solution of trifluoroacetic acid (0.33 g, 2.9 mmol) in anhydrous tetrahydrofuran (6 ml) under nitrogen was treated with l,l'-carbonyl diimidazole (0.50 g, 3.1 mmol). Some bubbling ensued, and the mixture was stirred for two hours, cooled on an ice bath, and combined under nitrogen with the residual amine prepared above (an additional 2 ml of tetrahydrofuran was used ~ to rinse the CDI adduct into the reaction vessel). The mixture was stirred at room temperature for 18h and concentrated in vacuo, then the residue was chromatographed on silica gel (eluted with 3% acetone/methylene chloride, then with 8%
acetone/methylene chloride) to afford, initially, syn-hexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenylmethyl-4-trifluoroacetylaminoazepine (0.27 g) followed by anti-WO 94/20062 21 S 7 ~ :12 PCT/US94/02283 hexahydro-3-(4-pllenylmethoxy)benzoylamino-1-phenylmethyl-4-trifluoroacetylaminoazepine (0.40 g). The total yield of trifluoroacetamides was 0.67 g (53%); the anti-isomer could be recrystallized from acetonitrile.
~ti-hexahydro-4-~methylamino)-3-(4-phenylmethoxy)benzoyl~ino-l-phenylmethylazep~ne An ice-cooled (5-C) solution of anti-hexahydro-3-(4-phenylmethoxy) benzoyl-amino-1-phenylmethyl-4-trifluoroacetylaminoazepine (0.20 g, 0.38 mmol) in anhydrous N,N-dimethylformamide (2 ml) under nitrogen was treated dropwise with 1.0 N potassium t-butoxide/tetrahydrofuran (Aldrich, 0.40 ml, 0.40 mmol), then stirred for 20 min at room temperature and recooled (5-C). Dimethyl sulfate (38 ~L, 0.40 mmol) was added via syringe, and stirring was continued at 5 C
for 3h. The solution was added to a rapidly stirred mixture of methylene chloride (10 ml) and saturated sodium bicarbonate (5 ml) and the organic layer was separated. The aqueous layer was extracted with methylene chloride (10 ml) and the combined organic solution was dried (Na2S04) and concentrated in vacuo.
The residue was chromatographed on silica gel (eluted successively with 2%, 3%, and 4% acetone/methylene chloride) to afford the methylated intermediate (0.15 g) as a viscous colorless oil. This was dissolved in reagent methanol (1.5 ml), treated with a solution of potassium hydroxide (0.25 g, 4.5 mmol) in water (0.25 ml), and stirred at room temperature~
for 2.5h. The solution was partially concentrated to remove most of the methanol, diluted with water (5 ml), and extracted with methylene chloride (2x15 ml). The combined organic extracts were dried (Na2S04) and concentrated in vacuo to afford anti-hexahydro-4-(methylamino)-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepine (0.105 g, 62%) as a colorless oil.
anti-~- t3,5-Bi~ (phenylmethoxy)-~-~2-carbophQnylmethoxy-6-phenylmethoxybenzoyl)benzoyl-N-methyl~mino]hexahydro-3-(~-phenylmethoxy)benzoyl~mino-l-phenylmethyl~zepine lC-I.r~uNv 620) A solution of 3,5-bis (phenylmethoxy) -4- (2-carbophenylmethoxy-6-phenylmethoxybenzoyl)benzoic acid (0.204 g, 0.30 mmol ) in anhydrous methylene chloride (1.0 ml ) was treated with N,N-dimethylformamide (2 drops), then with 2.0 N
oxalyl chloride/methylene chloride (Aldrich, 0.22 ml, 0.44 mmol ) and stirred for one hour under nitrogen . The solution was concentrated in vacuo, placed under high vacuum for 45 min, then dissolved in methylene chloride (2 ml ) and combined with anti-hexahydro-4- (methylamino) -3- (4-phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.10 g, 0.225 mmol). The mixture was treated with 1.0 N sodium hydroxide (1.0 ml) and stirred for two hours, then diluted with methylene chloride (15 ml ) and water (5 ml). The organic layer was separated and the aqueous solution was extracted with methylene chloride (2x15 ml). The combined organic solution was dried (Na2S0~) and concentrated in vacuo. The residue was chromatographed on silica gel (eluted successively with 5%, then 10% acetone/methylene chloride, then with 3% methanol/methylene chloride) to afford anti-4- [3,5-bis (phenylmethoxy) -4- (2 -carbophenylmethoxy-6-phenylmethoxybenz oyl ) benz oy 1 -N -methy 1 amino ] hexahydro- 3 - (4 -phenylmethoxy)benzoylamino-l-phenylmethyl-azepine (0.22g, 88%) as a viscous colorless oil.
anti-4-t4-~2-c~ 6-hy~.oa~L3nzoyl)-3,5-dihy~roxybenzoyl-N-methyl~mino]hexahydro-3-(~-hydroxybenzoyl~mino)~zepine, trifluoroacetic ~ci~ s~lt (CONPO~ND 539) A solution/suspension of anti-4- [3, 5-bis (phenylmethoxy ) -4 - ( 2 -carbophenylmethoxy-6-phenylmethoxybenzoyl ) benzoyl-N-methylamino ] hexahydro-3 - (4 -phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.22 g, 0.20 mmol ) in reagent ethanol (9 ml ) in a 25 ml 2 -neck round bottom flask was treated with trifluoroacetic acid (50 mg, 0.43 mmol), then with ethyl acetate (1 ml, for solubility) . Pearlman ' s WO 94l20062 21~ 7 412 PCT/US94/02283 catalyst (20% Pd(OH)2/C, so mg) was added, then the flask was quickly purged with nitrogen and fitted with a baloon valve and baloon containing hydrogen. The mixture was purged with hydrogen and kept under positive hydrogen pressure for 20h.
The flask was carefully evacuated of hydrogen and purged for several minutes with nitrogen. The solution was filtered through celite (wash filter cake with ethanol) and the filtrate was concentrated in vacuo and dissolved in N,N-dimethylformamide (1 ml). The solution was loaded onto a 41x250 mm C18 HPLC column and eluted as follows: A-0.1%
TFA/95:5 water:acetonitrile, B-acetonitrile, 100% A to 50:50 A:B over 60 min collected at 25 ml/min. The appropriate fractions were combined and partially concentrated in vacuo, then freeze-dried overnight to afford anti-4-[4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyl-N-methylamino]-hexahydro-3-(4-hydroxybenzoylamino)azepine, trifluoroacetic acid salt (101 mg, 68%) as a voluminous pale yellow solid; mp 285-295-C
(dec). Rf(6:1:1 n-BuOH/AcOH/H20) 0.45; IR (KBr): 1682, 1633, 1620 cm; H NMR (d6-DMSO) ~ 11.68 (br s, 2H), 10.07 (br s, lH), 9.92 + 9.851(s, lH), 8.95 + 8.751 (br s, 2H), 8.28 +
8.181(d, lH, J = 9 Hz), 7.67 + 7.581(d, 2H, J = 9 Hz), 7.20 -7.40 (m, 2H), 7.00 - 7.10 (m, lH), 6.78 - 6.85 (m, 2H), 6.00 (s, 2H), 4.40 - 4.80 (m, 2H), 3.00 - 3.40 (m, 4H), 2.85 +
2.771(s, 3H), 1.80 - 2.10 (m, 4H); mass spectrum(FAB): m/z 564.
Anal. Calcd. for C29H29N3Og-1~1(C2HO2F3)-3~0(H2O) C, 50.43; H, 4.90; N, 5.66. Found: C, 50.32; H, 4.74; N, 5.74.
21~7~12 ~094/2~62 - 237 - PCT~S94/02283 ~-R~-~- [ ( ( (3-hy~roxyc~rbonyl) -2-pyri~inyl) c~rbonyl) -3, 5-Oihy~roxybenzoyloxy] -3-R~ -hy~.o,.yL3nzami~0) perhy~roazepine trifluoroacetic ~ci~ s~lt (COIIrOu~v S~l) BnOOC O OBn BnO
1) (COC1)2 ,~\ ~
BnO CO2HCH2CI2 / DMF . o~oBnN
2) (~---N~OBn ~ NHb~OBn Ph 622 1 ) TFA
2) H2 Pd(OH)2 / C
EtOH
0~
(~,...N~O H
H
TFA
Trans-N-benzyl-~- t ~ ( (3-benzyloxyc~rbonyl) -2-pyridinyl ) c~rbonyl ) -3, 5-dibenzyloxybenzoyloxy] -3- ( ~-benzyloxybenzamido) perhydro~zepine (coMpo~rv 622 ) Carbonyldiimidizole (0.11 g, 0.65 mmole) was added to a solution of 4-[((3-benzyloxycarbonyl)-2-pyridinyl)carbonyl]-3,5-dibenzyloxybenzoic acid ( 0.25 g, 0.44 mmole) in 5 ml of 215 7 '1~ ~ PCT/US94102283 methylene chloride and the solution was stirred at room temperature for sixty minutes under nitrogen. The solution was added to a solution of 0.19 g (0.44 mmole) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyperhydroazepine, 0.12 ml triethylamine, and 5 mg DMAP in 8 ml of methylene chloride.
The solution was stirred at room temperature for twenty hours.
The solution was diluted with 30 ml of methylene chloride, washed with water, saturated brine and dried over magnesium sulfate. The solvent was removed in vacuo. The residue was chromatographed on silica gel eluting with a gradient of 5% -
10% - 20% ethyl acetate - hexane to yield 70 mg of a clear oil.
~.-R~-~- [~3-hy~roxycarbonyl)-2-pyridinyl)c~rbonyl)-3,5-dihydro~cybenzoylo~cy]-3-R~-~4-hy~roxybenzamido)perhydro~zepine trifluoro~cetic acid A solution of 0.070 g (0.071 mmole) of trans-N-benzyl-4-~(((3-benzyloxycarbonyl) -2-pyridinyl) carbonyl) -3, 5-dibenzyloxybenzoyloxy]-3-(4-benzyloxybenzamido)perhydroazepine in 8 ml of ethanol/methylene chloride (1:1) was treated with 10 L (0.142 mmole) of trifluoroacetic acid. The solution was stirred at room temperature for fifteen minutes. The solvent was evaporated and the ethanol/methylene chloride solvent was added twice more and evaporated in order to remove the excess trifluoroacetic acid. The residue was taken up in 10 ml of absolute ethanol and cooled to 0 C under nitrogen, and 0.030 g (0.025 mmole) of palladium hydroxide on carbon was added.
The reaction mixture was stirred under an atmosphere of hydrogen for twenty-four hours. The reaction mixture was filtered, evaporated and the residue was chromatographed on a 21 X 250 mm C18 column (solvent A: 95: 5 water/acetonitrile +
0.1% TFA; solvent B: 100% acetonitrile; gradient: 0 - 50% B
over 60 min., flow 15 ml/min). The pure fractions were pooled and evaporated to yield 0.010 g of a yellow powder, mp 198-205 C. Anal. Calcd for C27H25N30~-2H20-1.8TFA: C, 47.32; H, 4.00; N, 5.41. Found: C, 47.27; H, 3.86, N, 5.47.
W094/2~62 215 7 4 1 ~ PCT~S94/02283 ~+)-Trans-3-~-Hydroxy)benzamido-~ 3,5-~ihydroxy)-~-phenylcarbonyl]benzoyloxyperhydroazepine trifluoroacetic acid salt ~COMPOUND 544) OBn H~ O OOn O OH
Bn (~)-trans ~X~ ~F OH
1. (COC1)2 ,_~ N~f H2/Pd(OH)2 2. Et3N,DMAP ~ ~ (--~-` O
O OBn. Bn ~-~rans H CF3COOH
COMPOIJN`D 627 To a mixture of N-benzylated azepine intermediate (0.3 g, 0.697 mmol), Et3N (352.6 mg, 487 ~1, 3.485 mmol), and DMAP
(17.0 mg, 0.139 mmol) in CH2Cl2 (5 ml) was added a freshly made solution of benzophenone acid chloride (corresponding benzophenone acid: 336 mg, 0.766 mmol; oxalyl chloride: 2.0 M
solution in CH2C12, 0.697 ml, 1.394 mmol and cat. DMF) in CH2Cl2 (5 ml) at room temperature. The reaction mixture was stirred at room temperature overnight. Flash chromatography of the reaction mixture on silica gel using 3:2/Hexane:EtOAc as an eluent gave white solid product (469 mg, 71%).
CO~POUND 5~
The preceeding compound (300 mg, 0.315 mmol) in EtOAc:MeOH (1:1, 25 ml) was treated with CF3COOH (37.76 mg, W O 94t20062 2 ~ 5 7 ~ 12 PCTrJS94/02283 25.5 ~Ll, 0.33 mmol) and 20% Pd(OH)2 on activated carbon (150 mg, 50% on weight basis). The mixture was subjected to hydrogenolysis at 45 psi for 15 hr. The crude product after filtration and concentration was taken into DMF (0.5 ml) and purified on Cl8-HPLC column eluting with 596-50% acetonitrile in H2O containing 0.1% CF3COOH. The title compound was obtained as white powder (118 mg, 62%). m.p. 204-206 (dec)-C; 1HNMR
(DMSO-d6) ~ 8.42 (d, lH, NHCO), 7.58 (d, J = 7.5 Hz, 2H, ArH), 7.28-7.15 (m, 4H, ArH), 6.82 and 6.80 (s and s, 2H, ArH), 6.72 (d, J = 8.1 Hz, 2H, ArH), 5.18 (s, br, lH, CH--4), 4.45 (s, br, lH, CH-3), 3.30 and 3.10 (s and s, br, 4H, CH2N-2,7), 2.10-1.70 (m, 4H, CH2-5,6); IR (KBr) cm 1 3429, 1717, 1703, 1637, 1608, and 1509. Anal. Calcd. for C27H26N2O7 - 1.5 H2O - 1.0 TFA: C, 55.15; H, 4.79; N, 4.44. Found: C, 54.97; H, 5.08; N, 4.06.
_ WO94/2~K2 21~ 7 412 PCT~S94/02283 Trans-2- ~-Benzoyl-3, 5-dihy~roxybenzoyloxy) -1-hydroxybenzamido) cyclohexane ~COMPO~IND 545) Trans-2-(4-benzoyl-3,5-dibenzyloxybenzoyloxy)-1-(4-benzyloxybenzamido)-cyclohexane (172 mg, 231 ~mol) was dissolved in ethyl acetate (5 ml) and added to a stirring mixture of Pearlman's catalyst (Pd(OH) 2, 17 mg) in ethyl acetate (3 ml). The flask was evacuated under house vacuum and filled with H2(g) via balloon. After 3 h, additional Pearlman's catalyst (17 mg in 2 ml of EtOH) was added and left to stir overnight. TLC showed complete reaction (Starting material Rf=0.90, product Rf=0.41 in 50% EtOAc/hexanes.) The reaction mixture was filtered through Celite and concentrated to give the product as a yellow glass. The glass was triturated with water to give a pale yellow powder (122 mg, 86%): lH-NMR (DMSO, 300 MHz) ~ 1.27-1.57 (4H, m), 1.63-1.77 (2H, s), 1.84-1.89 (lH, m), 2.05-2.17 (lH, m), 4.05-4.18 (lH, m), 4.91-5.01 (lH, m), 6.75 (2H, d, J = 8 Hz), 6.97 (2H, s), 7.24-7.36 (2H, m), 7.48 (3H, t), 7.69 (2H, d, J = 8 Hz), 8.10 (lh, d, J = 9 Hz), 9.85 (lH, s), 9.95 (2H, s): 13C NMR (DMSO, 300 MHz) ~ 24.14, 24.52, 24.57, 31.20, 31.79, 51.73, 75.70, 107.68, 108.19, 114.98, 125.96, 126.29, 126.35, 128.35, 129.11, 129.15, 129.45, 132.08, 133.80, 137.24, 155.75, 160.29, 165.47, 166.13, 194.87. IR (KBr) cml 3360, 3271, 2943, 2858, 2360, 1708, 1659, 1610, 1541, 1507, 1450, 1424, 1369, 1347, 1278, 1240, 1177, 1106, 1047, 1011, 847, 771, 595. Anal. Calcd. for C27H2sNO7 - H20: C, 65.71; H, 5.51; N, 2.84. Found: C, 66.16: H, 5.49; N, 2.76.
W O 94/20062 215 7 ~12 PCTrUS94/02283 Anti-3-(~-benzyloxybenzami~o)-~-[3,5-dibenzyloxy-~-~3,~-dibenzyloxyphenylcarbonyl)benzoyloxy]-N-benzylperhy~roazepine ~CONPOUND 546) 3,5-Dibenzyloxy-4-[(3,4-dibenzyloxy)benzoyl]benzoic acid (500 mg, 0.768 mmol) was dissolved in anhydrous dichloromethane (6 ml). Anhydrous dimethylformamide (0.10 ml) was then added to the solution followed by oxalyl chloride (2 N in dichloromethane, 0.50 ml, 0.99 mmol). This solution was stirred for 1 h and then concentrated in vacuo. The resulting yellow oil was placed under high vacuum for a period of 1 h to make sure all of the excess oxalyl chloride was removed. The residue was dissolved in anhydrous dichloromethane (S ml), and a solution of dimethylaminopyridine (67 mg, 0.5S0 mmol), triethylamine, and anti-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine (SPC-103853, 215 mg, 0.500 mmol) in anhydrous dichloromethane (5 ml) was added under nitrogen. The reaction was stirred at room temperature for 1.5 h. At this point dichloromethane (100 ml) and sodium hydroxide (0.5 N in water, 30 ml) were added to the reaction. The aqueous and organic layers were separated and the organic phase was washed with brine (100 ml). The organic phase was isolated and dried with magnesium sulfate. The magnesium sulfate was filtered off, and the solution was concentrated in vacuo to yield a yellow solid.
The yellow solid was flash chromatographed on silica gel eluting with hexanes:ethyl acetate/9:1, 4:1, and 1:1 to yield a white solid of the title compound (400 mg, 75%): mp 65-C: lH
NMR (CDCl3) ~ 1.80 (m, 2H, CH2), 2.04 (m, 2H, CH2), 2.73 (m, 2H, NCH2), 3.01 (m, 2H, NCH2), 3.59 (d, J = 12.5 Hz, lH, NCH2Ph), 3.80 (d, J = 12.9 Hz, lH, NCH2Ph), 4.32 (m, lH, CH), 5.03 (s, 4H, CH2Ph), 5.12 (s, 2H, CH2Ph), 5.13 (s, 2H, CH2Ph), 5.22 (m, lH, CH), 5.24 (s, 2H, CH2Ph), 6.74 (d, J = 7.8 Hz, lH, NH), 6.86-7.56 (m, 39H, ArH); IR (KBr) cm 1580 (COO-), 1654 (CO).
Analysis calculated for C69H62N2O~: C, 77.95; H, 5.88; N, 2.63.
Found: C, 77.70; H, 5.99: N, 2.60.
W094/20062 21~ 7 412 PCT~S94/02283 Anti-~-[3~s-dihydroxy-~-~3~-dihydroxyphenylc~rbonyl)]
benzoyloxy-3-l~-hy~roxybenz~mido)perhydroazepine (COMPOUND 547) Anti-3-(4-benzyloxybenzamido)-4-[3,5-benzyloxy-4-(3,4-dibenzyloxyphenylcarbonyl)benzoyloxy]-N-benzylazepine (240mg, 0.23 mmol) and acetic acid were dissolved in methanol:ethyl acetate/2:1 in a 500 ml Parr bottle. Next, 5~ palladium on activated carbon (45 mg) was added under nitrogen. The reaction mixture was placed on a Parr hydrogenator for 3 h.
The mixture was then filtered over celite, and the filtrate was concentrated in vacuo to yield a yellow solid. The solid was flash chromatographed on silica gel eluting with dichloromethane:methanol/8:2 to yield a yellow solid of the title compound (57 mg, 48%): mp 176-C; H NMR (D6 DMSO) ~ 1.65 (m, lH, CH2), 1.78 (m, lH, CH2), 1.90 (2, 2H, CH2), 2.85 (m, 4H, N(CH2)2), 4.20 (m, lH, CH), 5.18 (m, lH, CH), 6.70-7.70 (m, 9H, ArH), 8.16 (d, J = 8.4 Hz, lH, NH); IR (KBr) cm1 1607 (CO), 1704 (COO-), 3431 (OH). Anal. calcd. for C27H26N2O8 H20: C, 59.50; H, 5.27; N, 5.14. Found: C, 59.54; H, 5.33; N, 4.93.
~1 L ~'7 W094/2~62 ~lJ J ~ PCT~S94102283 Trans-3-t3,5-Dihy~roxy-~-(2-hydroxy)phenylc~hQnyl]benzami~o-4-~-hydroxy)benzoyloxyperhy~ro~zepine ICOMr~u~v 548) To a solution of trans-N-benzyl-3-[3,5-dibenzyloxy-4-(2-benzyloxy)phenylcarbonyl]benzamido-4-(4-benzyloxy)-benzoyloxyazepine (65 mg, 0.068 mmol) in EtOAc/MeOH (5/15 ml) was introduced Pd(OH)z on carbon (35 mg of 20% Pearlman's catalyst). The reaction mixture was subjected to hydrogenolysis at 45 psi for 24 h at room temperature. The catalyst was filtered off through a pad of celite and washed with MeOH. The combined filtrate was concentrated to dryness and purified by flash column chromatography (SiO2: 60 ml, eluted with 10% to 20% MeOH in CH2C12). The title compound was obtained as a yellow solid (17 mg, 50%): mp 172-175-C, 1H NMR
(CD30D) ~: 7.65 (d, J = 8.7 Hz, 2H, ArH), 7.26 (td, lH, ArH), 7.10 (dd, lH, ArH), 6.74 (dd, lH, ArH), 6.60 (td, lH, ArH), 6.59 (d, J = 8.7 Hz, 2H, ArH), 6.50 (s, 2H, ArH), 5.03 (m, lH, CH-4), 1.19 (m, lH, CH-3), 2.95-2.70 (m, 4H, CH2N-2,7), 1.91-1.60 (m, 4H, CH2-5,6); IR (KBr) cm1 3434, 1700, 1623, 1610, 1542; low resolution FAB: (M + 1) 507.
_ W094/2~62 215 7 412 PCT~S94/02283 Tr~ns-3-t3,5-~ihy~roxy-4-~2-hy~roxyphenylra-honyl)]benz~mido-~-(4-hydroxy)benzoyloxy-N-isopropylperhydroazepine ~COMPOUND 549) To a solution of trans-N-benzyl-3-[3,5-dibenzyloxy-4-(2-benzyloxy)phenylcarbonyl]benzamido-4-(4-benzyloxy)-benzoyloxyperhydroazepine (190 mg, 0.199 mmol) in EtOAc-MeOH
(1:1, 30 ml) was introduced Pearlman's catalyst 20% Pd(OH)2 (70 mg). The mixture was subjected to hydrogenolysis at 50 psi for 17 hours. The catalyst was carefully filtered off through a pad of celite and washed with MeOH. The filtrate was concentrated and chromatographed on silica gel, eluting with 5%
MeOH in CH2C12. The title compound was obtained (500 mg, 50%);
the Parr bottle was contaminated with acetone and acid. All spectra ( H, C, APT, DEPT, MS) and CHN analysis fully support the structure. lH NMR (DMSO-d6) ~ 11.95 (s, brs, lH, OH), 10.25 (s, lH, OH), 10.00 (s, 2H, 20H), 8.20 (d, lH, NH), 7.78 (d, J
= 8.67 Hz, 2H, ArH), 753 (td, lH, ArH), 7.26 (dd, lH, ArH), 6.99 (d, lH, ArH), 6.87 (t, lH, ArH), 6.82 (d, J = 8.73 Hz, ArH), 6.74 (s, 2H, ArH), 5.01 (s, br, lH, CH-4), 4.18 (s, br, lH, CH-3), 2.90 (s, br, lH, CH(CH3)3), 2.68 (s, br, 4H, CH2N-2,7), 1.90 (s, br, 3H, CH2-6, CH-5), 1.63 (s, br, lH, CH-5), 1.00 (dd, 6H, 2CH3); 13C NMR (DMSO-d6 + D2O) ~ 55.02 (CH-(CH3)3), 18.66 and 18.40 (CH3)2CH); high resolution FAB M +
1: 549.2224; calculated for C30H32N2Oe: 549.2228. Anal. calcd.
for C30H32N2O~ 1.25 H2O: C, 63.09; H, 6.09; N, 4.90. Found: C, 63.06; H, 5.95; N, 4.66.
WOg4/2~62 21~ 7 4 ~ 2 PCT~S94/02283 --anti-4- ~3,5-Dihy~roxy-~- ~2-hy~roxybenzoyl)-benzoyl~mino)hexahy~ro-3-(4-hy~roxybenzoylamino)azepine, complex with water:acetonitrile ~1:1.7:0.3) (COMPOUND 550) ~8YNTg~8I8 OF COMPOUND 550) 8yn-3-Aminohexahy~ro-~-hydroxy-l-phenylmethylazepi~-2-one A solution of 3-acetylaminohexahydro-1-phenylmethyl-azepin-2,4-dione (0.82 g, 3.0 mmol) in absolute ethanol (15 ml) was treated with sodium borohydride (0.23 g, 6 mmol) and stirred for 30 min, then treated with water (5 ml) and concentrated in vacuo. The aqueous residue was extracted with methylene chloride (3 x 25 ml) and the combined organic extracts were dried (Na2SO~), concentrated in vacuo, and taken up in 2:1 ethanol/water (7.5 ml). Concentrated hydrochloric acid (2.5 ml) was added. The mixture was refluxed for 2 h and partially concentrated, then diluted with water (25 ml). The aqueous acidic mixture was extracted with ether (25 ml). The aqueous solution was basified with 30% sodium hydroxide and extracted with methylene chloride (3 x 40 ml). The combined methylene chloride extracts were washed with water (25 ml), dried (Na2SO~), and concentrated in vacuo to a yellow solid, which was recrystallized from ethyl acetate to afford syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.42 g, 60%) as a white solid.
8yn-3-Aminohexahydro-~-hydroxy-1-phenylmethylazepine A cooled (5-C) solution of lithium aluminum hydride/tetrahydrofuran (Aldrich, 1.0 N, 5.1 ml) under nitrogen was treated with syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.40 g, 1.7 mmol) in portions so that the pot temperature did not exceed 15-C. The mixture was refluxed for 6.5 h, cooled on an ice bath, and carefully treated with water (0.21 ml), 15~ sodium hydroxide (0.21 ml), and water (0.63 ml). The suspension was allowed to stir for 5 days (optimal time is 2-5 hours). The suspension was filtered, and the filtrate was concentrated in vacuo and chromatographed _ W094/2~62 215 7 412 PCT~S94/02283 on silica gel (eluted with 90:8:2 methylene chloride/methanol/
triethylamine). The appropriate fractions were concentrated in vacuo to afford syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.22 g, 58%) as a colorless oil.
8yn-~exahydro-~-hy~roxy-3-(4-phenylmethoxy)benzoyl~mino-1-phenylmethyl~zepine A solution of 4-benzyloxybenzoic acid (0.183 g, 0.8 mmol) in anhydrous tetrahydrofuran (2 ml) and N,N-dimethylformamide (0.5 ml) was treated with N,N'-carbonyldiimidazole (O.lS g, 0.9 mmol) and stirred at room temperature for 1.5h. The solution was treated with syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.20 g, 0.9 mmol) in anhydrous tetrahydrofuran (1 ml). The mixture was stirred for 18 h, then concentrated in vacuo. The residue was taken up in lN sodium carbonate (20 ml), and the aqueous mixture was extracted with toluene (2x25 ml) cont~ining a little 2-propanol. The combined organic extracts were dried (Na2S0~) and the concentrated residue was flash chromatographed on silica gel (eluted with 3:1 ethyl acetate/hexane) to afford syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.17 g, 50%) as a viscous oil.
syn-~-Aminohex~hydro-3-(~-phenylmethoxy)benzoyl~mino-1-phenylmethylazepine A solution of syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)-benzoylamino-l-phenylmethylazepine(0.645g,1.5 mmol) in anhydrous tetrahydrofuran (3 ml) under nitrogen was cooled (ice bath, 5-C) and treated with triphenylphosphine (0.495 g, 1.8 mmol) and diethylazodicarboxylate (0.313 g, 1.8 mmol). The mixture was then treated dropwise over 15 min with a solution of diphenylphosphoryl azide (0.495 g, 1.8 mmol) in anhydrous tetrahydrofuran (3 ml). The reaction was stirred at room temperature for 18 h, then concentrated in vacuo, dissolved in a little methylene chloride, and passed through a short column of silica gel (eluted with 10% acetone/methylene chloride). The early fractions containing chromophoric W094/2~62 215 7 ~1~ PCT~S94/02283 material were concentrated in vacuo and dissolved in ethanol/acetic acid/water (6:1:1, 12 ml), then treated with zinc powder (0.50 g, 7.5 mmol). After 30 min, the mixture was filtered and the filtrate was concentrated in vacuo. The residue was taken up in 1.0 N sodium hydroxide (40 ml), and the aqueous mixture was extracted with methylene chloride (3 x 35 ml). The combined extracts were dried (Na2SO4) and concentrated in vacuo to afford crude syn-4-aminohexahydro-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepine (0.76 g).
syn-4-~3,5-Bis~phenylmethoxy)-~-~2-phenylmethoxybenzoyl))-benzoyl~minohexahydro-3-~-phenylmethoxy)benzoyl~mino-1-phenylmethylazepine ~COMPOUND 628) A solution of 2,2',6-tribenzyloxybenzophenone-4-carboxylic acid (0.545 g, 1.0 mmol) in anhydrous methylene chloride (3 ml) and N,N-dimethylformamide (0.1 ml) under nitrogen was treated with 2.0 N oxalyl chloride/methylene chloride (0.7 ml, 1.4 mmol) and stirred for one hour. The solution was concentrated in vacuo to a yellow solid, which was kept under high vaccuum for one hour. A solution of crude syn-4-aminohexahydro-3-(4-phenylmethoxy)-benzoylamino-1-phenylmethylazepine (0.76 g, from above) in methylene chloride (5 ml) was added to the acid chloride above, followed by 1.0 N
sodium hydroxide (4 ml). The mixture was stirred for 1.5 h, diluted with methylene chloride (15 ml) and separated. The aqueous layer was extracted with methylene chloride (15 ml) and the combined organic solution was washed with saturated aqueous sodium chloride, dried (Na2SO~), and concentrated in vacuo. The residue was chromatographed on silica gel (eluted initially with 2.5% acetone/methylene chloride, then with 7.5%
acetone/methylene chloride) to afford syn-4-(3,5-bis(phenylmethoxy)-4-(2-phenylmethoxy-benzoyl))-benzoylaminohexahydro-3-(4-phenylmethoxy)-benzoylamino-1-phenylmethylazepine (0.52 g, 54%) as an opaque gum.
_ WO 94/20062 21 S 412 PCT/US94/02283 syn~ 3, 5-Dihydroxy-~- ~ 2-hydroxybenzoyl ) -benzoylamino)hexahy~ro-3-14-hydroxybenzoylamino)azepine, complox with water:acetonitril~ ~1:1.7:0.3) (COMPO~ID 550) A solution of syn-4- (3,5-bis (phenylmethoxy) -4- (2-phenylmethoxybenzoyl ) ) benzoylaminohexahydro-3 - (4 -phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.39 g, 0.41 mmol) in ethanol/ethyl acetate (1: 1, 30 ml) was placed in a Parr bottle and treated (under nitrogen) with Pearlman ' s catalyst (Aldrich, 150 mg), then subjected to hydrogenation in a Parr apparatus for 18 h at 48-50 psi. The reaction mixture was carefully purged of hydrogen and the solution was filtered through celite (care taken not to let filter cake dry). The filtrate was concentrated in vacuo to afford crude material, which was chromatographed on silica gel (eluted with 2 :1 methylene chloride/isopropanol) to give syn-4- (3,5-dihydroxy-4-( 2 -hydroxybenzoyl ) benzoylamino ) hexahydro-3 - ( 4 -hydroxybenzoylamino)azepine, complex with water:acetonitrile (1:1.7:0.3) (0.055 g, 27%) as a yellow solid, which was triturated with acetonitrile to afford a yellow powder: mp 210 - 215-C; Rf (2: 1 methylene chloride/isopropanol on silica) 0.50; IR (KBr) 1624 cm 1; lH NMR (d6-DMSO) ~ 8.33 (d, lH, J = 7 Hz), 7.97 (d, lH, J = 7 Hz), 7.73 (d, 2H, J = 9 Hz), 7.55 (m, lH), 7.30 (m, lH), 7.00 (m, lH), 6.89 (m, lH), 6.84 (s, 2H), 6.82 (d, 2H, J = 9 Hz), 4.25 - 4.40 (m, 2H), 3.00 - 3.10 (m, 2H), 2.80 - 3.00 (m, 2H), 1.85 - 1.95 (m, lH), 1.65 - 1.80 (m, 3H). Anal. calcd. for C2~H27N307 1.7H20 0.3(CH3CN): C, 60.44;
H, 5.75; N, 8.43. Found: C, 60.34; H, 5.56; N, 8.42.
W094/20062 215 7 41~ PCT~S94/02283 Hexahydro-3-(~-phenylmethoxy)benzoylamino-1-phenylmethyl-azepin-~-one (COMPOUND 551) A 25 ml 3-neck round bottom flask under nitrogen was charged with 2.0 N oxalyl chloride/methylene chloride (Aldrich, 1.125 ml, 2.25 mmol), diluted with anhydrous methylene chloride (2 ml), cooled (-65-C), and treated dropwise with anhydrous dimethylsulfoxide (0.35 g, 4.5 mmol) in anhydrous methylene chloride (1.2 ml) at a rate to keep the pot temperature below -60-C. The mixture was stirred at -65 + 5C for 30 min, then treated dropwise with a solution of syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.645 g, 1.5 mmol) in anhydrous methylene chloride (1.5 ml) at a rate to keep the pot temperature below -55-C. The mixture was stirred at 55 + 5-C for 2 h, then treated dropwise with triethylamine (1.5 ml), warmed to room temperature over 1 h, and diluted with methylene chloride (10 ml). The organic solution was washed with water (10 ml), saturated aqueous sodium bicarbonate (10 ml), dried (Na2S0~), and concentrated in vacuo. The residue was chromatographed on silica gel (eluted with 5% acetone/methylene chloride) to afford hexahydro-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepin-4-one (0.53 g, 82%) as a viscous colorless oil.
~-Aminohexahydro-3-(~-phenylmethoxy)benzoylamino-1-phenyl-methylazepine, 1:1 mixture of syn and anti i~omers A solution of hexahydro-3-(4-phenylmethoxy)-benzoylamino-l-phenylmethylazepin-4-one (0.37 g, 0.86 mmol) in ethanol (5 ml) was treated with hydroxylamine hydrochloride (80 mg, 1.15 mmol) and 25% methanolic sodium methoxide (Aldrich, 70 mg, 0.32 mmol) and heated to 50-C for 1 h. The mixture was cooled to room temperature, treated with additional 25% sodium methoxide (0.18 g, 0.83 mmol), then stirred for 10 min and partially concentrated in vacuo. The residue was taken up in 0.5 N sodium hydroxide (5 ml) and extracted with methylene chloride (3 x 15 ml). The combined organic extracts were washed with saturated aqueous sodium chloride, dried (Na2SO4), and concentrated in vacuo. This residue was dissolved in 95%
W094/20062 2 1 ~ 7 4 ~ ~ PCT/US94/022~
ethanol (20 ml), treated with Raney Nickel (Aldrich, 1/4 teaspoon) in a Parr bottle, and subjected to hydrogenation at 30 psi for 2.25 h (more time needed). The solution was carefully evacuated of hydrogen, filtered through celite (do not allow to dry), and the filtrate was concentrated in vacuo and chromatographed on silica gel (eluted first with 15%
methanol/methylene chloride, then with 25%, and finally with 33%) to afford 4-aminohexahydro-3-(4-phenylmethoxy)benzoyl-amino-l-phenylmethylazepine, 1:1 mixture of syn and anti isomers (0.15 g, 40%) as a viscous colorless oil which was stored under nitrogen.
~nt~-4-~3,5-B$~phenylmethoxy)~ 2-phenylmethoxybenzoyl))-benzoyl~minohexahydro-3-(~-phenylmQthoxy)benzoyl~mino-l-phenylmethylazepine ~COMPO~ND 629) A solution of 2,6,2'-tribenzyloxybenzophenone-4-carboxylic acid (0.245 g, 0.45 mmol) in anhydrous methylene chloride (1.5 ml) under nitrogen was treated with N,N-dimethylformamide (3 drops), then with 2.0 N oxalyl chloride/methylene chloride (0.30 ml, 0.60 mmol). The vigorous bubbling soon subsided and the mixture was stirred for 1 h, concentrated in vacuo, and kept under high vacuum for 1 h to ensure removal of excess oxalyl chloride. The acid chloride was dissolved in methylene chloride (2.5 ml) and treated with 4-aminohexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine, 1:1 mixture of syn and anti isomers (0.14 g, 0.325 mmol) followed by 1.0 N sodium hydroxide (1.5 ml).
The biphasic mixture was rapidly stirred for 2 h and separated.
The aqueous layer was extracted with methylene chloride (2 x 7 ml), and the combined organic extracts and organic layer were washed with saturated aqueous sodium chloride (10 ml), dried (Na2SO~), and concentrated in vacuo. Flash chromatography on silica gel (eluted first with 3% acetone/methylene chloride, then with 7%, and finally with 10%) afforded anti-4-(3,5-bis(phenylmethoxy)-4-(2-phenylmethoxy-benzoyl))-benzoylaminohexahydro-3-(4-phenylmethoxy)benzoylamino-1-WO 94/20062 215 7 il ~ 2 PCT/US94/02283 phenylmethylazepine (0.13 g, 42%) as a colorless foam, followedby the syn isomer (0.13 g, 42%) .
~nti-4-~3,5-Dihydroxy-~-~2-hydroxybenzoyl)benzoyl~mino)hexa-hydro-3-~-hydroxybenzoylamino)azepine, complex with water:2-prop~nol ~1:1.5:0.~) ~COMPOUND 551) A solution of syn-4- (3,5-bis (phenylmethoxy) -4- (2-phenylmethoxybenzoyl) ) benzoylaminohexahydro-3- (4-phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.13 g, 0.136 mmol) in ethanol/ethyl acetate (1: 1, 25 ml) was placed in a Parr bottle and treated (under nitrogen) with Pearlman ' s catalyst (Aldrich, 120 mg), then subjected to hydrogenation in a Parr apparatus for 18 h at 50 psi. The reaction mixture was carefully purged of hydrogen and the solution was filtered through celite (care taken not to let filter cake dry). The filtrate was concentrated in vacuo to afford crude material, which was chromatographed on silica gel (eluted with 1: 1 methanol/methylene chloride). This concentrated product was dissolved in 2-propanol, filtered to remove inorganic material, re-concentrated in vacuo, and triturated from ether to afford anti-4- ( 3, 5-dihy~roxy-4- ( 2 -hydroxybenzoyl ) -benzoylamino) hexahydro-3- (4-hydroxybenzoylamino) azepine complex with water:2-propanol (1:1.5:0.4) (56 mg, 75%) as a yellow powder: mp 207 - 210 C; Rf (2: 1 methylene chloride/2-propanol on silica) 0.20: IR (KBr) cm 1 1639, 1607; H NMR (d6-DMSO) 8.32 (d, lH, J = 7 Hz), 7.96 (d, lH, J = 7 Hz), 7.66 (d, 2H, J
= 9 Hz), 7.52 (m, lH), 7.27 (m, lH), 6.97 (m, lH), 6.86 (m, lH), 6.77 (d, 2H, J = 9 Hz), 6.72 (s, 2H), 4.00 - 4.15 (m, 2H), 2.60 - 3.00 (m, 4H), 1.50 - 1.90 (m, 4H) . Anal . calcd. for C2~Hz~N307 1-5 H20 0.4 (C3H80): C, 60.85; H, 6.01; N, 7.55.
Found: C, 60.68; H, 5.68; N, 7.22.
~15~ 2 - PCT~S94/0~R~
~ Anti-3-~ minobenzam~do)-~-[3,5-dihy~roYy-~-(2-hy~roxyphenylcarbonyl)]benzyloxyperhy~roazepine (COMPOUND552) OE~ 8110 OH HO
OH H ` O~ ~ o (';;)-- ,C, ~ N2 ~ C~ NOz -- (;~-- ~ C~ NH2 3,5-dibenzyloxy-4-~(2-benzyloxybenzoyl]benzoic acid (218 mg, 0.40 mmol) was dissolved in anhydrous dichloromethane (2.0 ml), and dimethylformamide (0.05 ml). The reaction was placed under nitrogen, and oxalyl chloride (0.3 ml, 2M in dichloromethane) was slowly added via a syringe. The reaction was stirred at room temperature for 1 h and then concentrated in vacuo and placed under high vacuum for 1 h. The residue was redissolved in anhydrous dichloromethane (2 ml) and stirred under nitrogen. Next, a solution of (+)-anti-3-(4-nitrobenzamido)-4-hydroxybenzylperhydroazepine (120 mg, 0.32 mmol) and dimethylaminopyridine (44 mg,0.36 mmol) in triethylamine (0.9 ml) and anhydrous dichloromethane (3 ml) were added. The reaction was then stirred for 1 h at room temperature. At this point, dichloromethane (50 ml) and 0.5 N
sodium hydroxide (20 ml) were added, and the reaction was transferred to a separatory funnel. The aqueous layer was removed, and brine was added. The organic layer was then isolated, dried over sodium sulfate, filtered, and concentrated in vacuo to yield a yellow oil. The oil was flash chromatographed eluting with hexane:ethyl acetate/4:1. Pure fractions of the major product were concentrated in vacuo to yield 180 mg of white solid (COMPOUND 636). The solid was then dissolved in methanol:ethyl acetate/2:1 (30 ml) and acetic acid ~094/20062 2 1 ~ 7 4 1 2 PCT~S94/02283 (0.2 ml) in a 500 ml Parr bottle along with 5% palladium on activated carbon (70 mg). The Parr bottle was then placed on the Parr hydrogenator for 24 h. The reaction mixture was filtered to remove the catalyst, and then concentrated in vacuo to yield a yellow solid. The solid was flash chromatographed eluting with dichoromethane:methanol/9:1 to yield a yellow solid of the title compound (60 mg, 61%): mp 175 - 185-C; NMR
(CD30D) consistant with structure; IR (RBr) cm1 3389 (OH); 2361 (alkyl); 1704 (ester); 1625 (ketone). Anal. calcd. for C27H27N3O7 1.0 H2O 1.0 acetic acid: C, 59.69; H, 5.70; N, 7.20. Found: C, 59.94; H, 5.70; N, 7.31.
~09~/20062 21~7412 _ PCT~S94/0 ~ + ) -Anti-3- ( ~- f luoro~^n~mi~o)-4-t3,5-dihydroxy-~-(2-hy~roxy-phenylc~rbonyl)]benzyloxyperhydroazepine (COMPOUND 53) OBn i3nO OH HO
0,.~~ o ,~_,0,~
o~ F _ (;;)-- C~ F (;;~_ ~H OH F
3,5-Dibenzyloxy-4-t(2-benzyloxybenzoyl]benzoic acid (545 mg, 1.0 mmol) was dissolved in anhydrous dichloromethane (5.0 ml), and dimethylformamide (0.05 ml). The reaction was placed under nitrogen, and oxalyl chloride (1.0 ml, 2 M in dichloromethane) was slowly added via a syringe. The reaction was stirred at room temperature for 2 h and then concentrated in vacuo and placed under high vacuum for 1.5 h. The residue was redissolved in anhydrous dichloromethane (3 ml) and stirred under nitrogen. Next, a solution of (+)-anti-3-(4-fluorobenzamido)-4-hydroxy-N-benzylperhydroazepine (300 mg, 0.93 mmol) and dimethylaminopyridine (125 mg, 1.02 mmol) in triethylamine (0.9 ml) and anhydrous dichloromethane (2 ml) were added. The reaction was then stirred for 20 min at room temperature. At this point, dichloromethane (50 ml) and 0.5 N
sodium hydroxide (30 ml) were added, and the reaction was transfered to a separatory funnel. The aqueous layer was removed, and brine was added. The organic layer was then isolated, dried over sodium sulfate, filtered, and concentrated in vacuo to yield a light brown foam. The foam was flash chromatographed eluting with hexane:ethyl acetate/2:1. Pure fractions of the major product were concentrated in vacuo to yield 440 mg of white solid (COMPOUND 631). 260 mg of the W094/20062 2 1 5 7 4 ~ ~
PCT~S94/02283 solid was dissolved in methanol:ethyl acetate/5:1 (30 ml) and acetic acid (0.2 ml) in a 500 ml Parr bottle along with 5%
palladium on activated carbon (100 mg). The Parr bottle was then placed on the Parr hydrogenator for 4.5 h. The reaction mixture was filtered to remove the catalyst, and then concentrated in vacuo to yield a yellow solid. The solid was flash chromatographed eluting with dichoromethane:methanol/8:2 to yield a yellow solid of the title compound (110 mg, 72%):
mp 189-C; IR (KBr) cm1 3349 (OH); 2362 (alkyl); 1704 (ester);
1626 (ketone). Anal. calcd. for C27H25N2O7F 1.0 H2O 1.0 acetic acid: C, 59.38; H, 5.33; N, 4.78. Found: C, 59.09; H, 5.00; N, 4.71.
W094/20062 2 1 5 7 ~ 1 2 - PCT~S94/022 Tr~ns-3-~4-hy~,ox~z~m~do)-~-t~-(2-hy~oh~ 6-hy~ c~rhonyl-benzoyl)-3,5-~ihy~roxybenzoyloxyl]perhy~rothiepine (COMPOUND
55~) N~NHa 13~OH m~ DMF O~OTBDMS )O~H~ ~OH
HO~OH Ms Cl. EI~N ~AsO~OM~ L~MS EI~N TBDIA5~N, H NH~ 1) ~~ CO~ OBn ~
I.JAIH~THF ~ CDI, THF H ~ COCI
H~ ~N~
\_~S 2)lNN OH H~O ~S El~N DM~P,CH~
OBn OE~n H~ Pd(OHb~ ~(OH OH
l~nO~_ H ~ ~ OH THF HO ~_ ~N~ O ~ O
S
A mixture of 1,3-cyclohexadienemonoepoxide (9.61 g, 100 mmol, for preparation see: J. K. Crandall et àl., J. Org.
Chem., 1968, 33, 423), NaN3 (26 g, 400 mmol), NH4Cl (10.7 g, 200 W094/20062 215 ~ PCT~S94/02283 mmol), MeOH (300 ml), and H2O (50 ml) was stirred at 65 C for 16h. The resultant slurry was evaporated to remove all the volatile materials and the residue was treated with 1 N NaOH
(100 ml), and extracted with CH2C12 (3 x 60 ml). The combined CH2C12 extracts were washed with H20 (2 x 100 ml), dried (MgSO4), evaporated, and chromatographed on a SiO2 column (Et2O:hexane = 1:4) to give a colorless oil (7.308 g, 53%).
A mixture of the product of the previous reaction (7.23 g, 52 mmol), TBDMS-Cl (8.227 g, 55 mmol), and imidazole (5.308 g, 78 mmol) in N,N-dimethylformamide (50 ml) was stirred at room temperature for 16h. Et2O (250 ml) was added and the resultant mixture was washed with H2O (5 x 100 ml), dried (MgSO~), and evaporated to give a colorless oil (12.76 g, 97%) which was used in the next step without further purification.
A solution of oil (7.63 g, 30 mmol) in MeOH (150 ml) and CH2C12 (30 ml) was cooled to -78-C, stirred, and treated with a mixture of air and 03 until a blue color persisted (ca.
1.5h). Excess O3 was removed by bubbling N2 through the solution, and then NaBH~ (4.54 g, 120 mmol) was added. After being stirred at -78-C for 30 min. followed by 30 min. at room temperature the resultant mixture was poured into 1 N HCl (100 ml) and then evaporated to remove the volatile components. The aqueous residue was extracted with Et2O (4 x 50 ml), and the combined ether extracts were washed with HzO (100 ml), brine (100 ml), dried (MgSO~), evaporated, and crystallized from Et2O:hexane to give a white solid (6.772 g, 78%).
Methanesulfonyl chloride (2.67 ml, 34 mmol) was added dropwise to a stirred solution of the borohydride reduction product (4.01 g, 14 mmol) and Et3N (S.76 ml, 41 mmol) in CH2Clz (40 ml) at 5-C. After 30 min. the cooling bath was removed and stirring was continued for 16h. The resultant mixture was diluted with CH2C12 (20 ml), washed with 1 N HCl (3 x 30 ml), H2O (2 x 30 ml), brine (2 x 30 ml), dried (MgSO4), and evaporated. The residue was chromatographed (SiO2, Et2O:hexane = 1:1 followed by Et2O:hexane:CH2Cl2 = 1:1:0.25) to give a pale yellow oil (5.885 g, 96%).
UO9~/20062 2 1 5 7 ~ 1 2 PCT~S94/02283 A mixture of the product of the previous reaction (2.24 g, 5 mmol), lithium sulfide (253 mg, 5.5 mmol), and Et3N
(2.09 ml, 15 mmol) in EtOH (100 ml) was stirred under reflux for 16h. The resultant mixture was evaporated and the residue was chromatographed (SiO2, Et2O:hexane = 1:30) to give a colorless oil (515 mg, 36%).
A solution of LiAlH4 in Et2O (1 M, 3.58 ml) was added to the colorless oil from the reaction above (515 mg, 1.79 mmol) in Et2O (9 ml) and the mixture was stirred at room temperature for 2h. 5N aq NaOH (5 ml) was added cautiously and stirring was continued at room temperature for 16h. The phases were separated and the aqueous phase was saturated with NaCl and extracted with CH2C12 (3 x 10 ml). The combined organic layers were dried (MgSO4), and evaporated to give a colorless oil which solidified on standing (250 mg, 95%). This material was used in the next step without further purification.
A mixture of 4-benzyloxybenzoic acid (427 mg, 1.87 mmol) and l,l'-carbonyldiimidazole (302 mg, 1.87 mmol) in THF
(5 ml) was stirred at room temperature for 2h and then the product of the previous reaction (250 mg, 1.70 mmol) in THF (5 ml) was added. The mixture was stirred at room temperature for 36h, evaporated, dissolved in CH2C12 (30 ml), washed with H2O, and dried (MgSO~). The resultant solution was diluted with hexane (10 ml) and rotaevaporated at 0-C to ca. 10 ml. The white precipitate was collected, dried under vacuum, and dissolved in a mixture of MeOH (10 ml) and THF (8 ml). 1 N aq.
NaOH (1 ml) was added and the mixture was stirred at room temperature for 4h. The volatile components were evaporated and the residue was dissolved in CH2C12 (25 ml), washed with H2O
(3 x 5 ml), dried (MgSO4), and filtered. The filtrate was diluted with hexane (lO ml) and rotaevaporated at 0 C to ca. 5 ml. The precipitate was collected, washed with hexane, and dried under vacuum to give a white powder (382 mg, 67%).
To a solution of 4-(2-benzyloxy-6-benzyloxycarbonyl-benzoyl)-3,5-dibenzyloxybenzoic acid (305 mg, 0.45 mmol) and a drop of N,N-dimethylformamide in CH2C12 (1.3 ml) at 5C was w094~20062215 7 ~12 PCT/US94/02283 added a solution of oxalyl chloride in CH2Cl2 (2 M, 0.25 ml).
The resultant mixture was stirred at room temperature for 2h and then evaporated. The residue was dried under vacuum for 2h, dissolved in CH2C12 (0.8 ml), and added to a mixture of the above intermediate (143 mg, 0.4 mmol), Et3N (61 mg, 0.6 mmol), and 4-N,N- dimethylaminopyridine (5 mg, 0.04 mmol) in CH2Cl2 (1.2 ml). The resultant solution was stirred at room temperature for 17h, diluted with CH2C12 (10 ml), washed with H2O (3 x 5 ml), dried (MgSO~), and evaporated. The residue was chromatographed (SiO2, Et2O:hexane:CH2Cl2 = 1:1:0.5) to give a colorless oil (347 mg, 76%).
This intermediate (105 mg, 0.1 mmol), Pd(OH)2 on carbon (20 wt%, contains > 50% moist, 281 mg, 0.2 mmol), THF (1 ml), and MeOH (1 ml) was stirred and treated with 1 atm H2 at room temperature for 40h. The resultant mixture was filtered through a Florisil pad and the Florisil pad was washed with MeOH (15 ml). The combined filtrate and wash were evaporated to give a yellow solid (25 mg, 35%) (COMPOUND 554). IR (KBr, cm ): 1706, 1689, 1633.
1, 1-Dioxo-trans-3- ~ ~-hydroxybenz~mi~o ) -~- t ~.- ( 2-hy~roxy-6-hydroxycarbonylbenzoyl ) -3, 5-~lihy~roxybenzoyloxyl ] -perhydrothiepine ~COMPO~JND 555) ~o ~o CH,CO,H ~_ ~ ~CH,CO,H. ct'.,a, 11l1 ~ OH OH
H,. Pd~OH), C
C~so o Figure AA
Peroxyacetic acid (32 wt% in acetic acid, 37 mg, 0.155 mmol) was added to a solution of Compound 632 (75 mg, 0. 074 mmol, see Compound 554 for preparation) in CH2Cl2 (0.7 ml). The resultant mixture was stirred at room temperature for lh, diluted with CH2C12 (10 ml), and washed with sat. aq. K2CO3 ( 3 x 5 ml). The organic layer was dried (MgSO4) and evaporated to give a white solid which was recrystallized from hot EtOAc (10 ml, contains ca. 2 ml of THF and 3 ml of hexane) to give a white powder (61 mg, 79~) (COMPOUND 633).
21~7412 ~094/20062 PCT~S94/02283 A mixture of Compound 633 (61 mg, 0.058 mmol), Pd(OH)2 on carbon (20 wt%, contains > 50% moist, 8 mg, 0.0058 mmol), THF (1 ml), and MeOH (1 ml) was stirred and treated with 1 atm H2 at room temperature for 3 hr. The resultant mixture was filtered through Florisil and the filtrate was evaporated to give the title compound as a yellow solid (33 mg, 95~). IR
(KBr, cm ): 1718, 1686, 1635.
_ WO9~/20062 21~7~
Tr~ns-4-(4-(2-c~rboxy-6-hydroxybenzoyl)benzoyloxy)-3-(4-hy~roxybenz~mi~o)perhydro~zepine Trifluoro~cetic Aci~ ~lt Hy~r~te ~COMPOUND 556) OBn ( CO2H
~ t~ BuU ~ PDC, DMF
MeO~OMeO~SJ oan MeO~I OBn CO2Bn BnB- ~ KOH, DMF
KDCMO~3 MeO ~ OBn o CO2Bn BnO~
)~\ 1) (coa)2 ~, OBn "~/ 2) IPr2NEI, DMAP, ~/
HO2C OBn OH O ~~
(~ . N ~ OBn ~ H~ OBn Bn Bn 6~
HoO
H2 ~ OH
Pd(OHh . N~ OH
H
W094/20062 215 7 ~12 PCT~S94/02283 Methyl-4-(6-Benzyloxy-2-hy~roxymethylbenzoyl)benzoate To a solution of 1.07 g (5.00 mmol) of 3-benzyloxybenzyl alcohol in 15 ml of toluene at -5CC under an atmosphere of nitrogen was added 5.8 ml (12.2 mmol) of a 2.1 M solution of butyllithium in hexanes over 15 min. The solution was stirred at -5C for 6 h, after which it was cooled to -78-C, and a solution of 1.00 g (5.03 mmol) of 4-(methoxycarbonyl)benzoyl chloride in 5 ml of tetrahydrofuran was added. The mixture was stirred for 1 h, after which it was poured onto 200 ml of ether and 100 ml of saturated aqueous ammonium chloride, and this mixture was stirred for 10 min.
The layers were separated, and the organic phase was washed with saturated aqueous sodium bicarbonate and brine, dried over magnesium sulfate, and evaporated to give the crude product.
Flash chromatography on silica gel eluting with 3/1 ethyl acetate -hexane afforded 0.68 g (36~) of the title compound as a white solid, which was carried on to the next step.
Methyl-~-~6-Benzyloxy-2-ca,~6~benzoyl)benzoate To a solution of 0.63 g (1.7 mmol) of methyl 4-(6-benzyloxy-2-hydroxymethylbenzoyl)benzoate in 20 ml of dimethylformamide was added 4.41 g (11.7 mmol) of pyridinium dichromate. The solution was stirred at room temperature under a nitrogen atmosphere for 4 days, after which it was poured onto 300 ml of ether and washed with 200 ml of water, 150 ml of 2 M HCl, and 150 ml of brine, and dried over magnesium sulfate.
Evaporation of the solvent afforded 0.47 g (72%) of the crude product. This material was sufficiently pure for further use and was carried directly to the next step.
~ethyl-4-~6-Benzyloxy-2-~benzyloxyc~rbonyl)benzoyl)benzoate To a c ~ution of 0.47 g (1.2 mmol) of methyl 4-(6-benzyloxy-2-carboxybenzoyl)benzoate in 20 ml of dry dimethylformamide was added 501 mg (3.62 mmol) of potassium carbonate and 0.158 ml (227 mg, 1.32 mmol) of benzyl bromide.
The solution was stirred at room temperature under a nitrogen atmosphere for 18 h. The mixture was then poured onto 300 ml WO9-1/20062 215 7 ~12 PCT~S94/02~83 of ether and washed with two 200 ml portions of water and then with 150 ml of brine, and dried over magnesium sulfate.
Evaporation of the solvent afforded 0.57 g of the crude product, which was chromatographed on silica gel, eluting with 3/1 hexane-ethyl acetate to give 0.32 g (54%) of the title compound as a colorless oil. This material was used directly in the next step.
~-(6-Benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoic Acid A solution of 0.301 g (0.614 mmol) of methyl 4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoate in 7 ml of DMF
was treated with 0.337 ml of a 2 M aqueous solution of potassium hydroxide under an atmosphere of nitrogen for 20 h.
The mixture was then poured onto 100 ml of ethyl acetate and washed with 60 ml each of 0.2N HCl, water, and brine. The organic extracts were dried over magnesium sulfate and evaporated to give 0.32 g of the crude product, which was chromatographed on a 41 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA solvent B: 100% acetonitrile:
gradient: 0-100% B over 60 min, flow: 25 ml/min). The pure fractions were pooled and evaporated and then lyophilized from water to give 70 mg of the title compound as a white solid, which was carried on as is to the next step.
Tr~ns-N-Benzyl~ -(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl) benzoyloxy)-3-(~-benzyloxybenz~mido)perhydroazepine A solution of 68 mg (0.143 mmol) of 4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoic acid in 5 ml of methylene chloride containing a trace (approximately 1 ~L) of dimethylformamide was cooled to O-C. A 2.0 M solution of oxalyl chloride (0.11 ml, 0.22 mmol) was added, and the mixture was stirred under a nitrogen atmosphere for 2 h. An additional 0.17 ml of oxalyl chloride was added, and the mixture was stirred for an additional 2 h. The reaction mixture was evaporated, and the residue was evaporated twice from 10 ml of methylene chloride. The residue was dissolved in 3 ml of methylene chloride, and was added to a solution of 69.6 mg 21~1 2 (0.162 mmol) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine, 29.8 ,uL (0.17 mmol) of diisopropylethylamine, and 5.4 mg of DMAP in 5 ml of methylene chloride at O-C. The mixture was stirred at room temperature under a nitrogen atmosphere for 17 h, after which it was diluted with 30 ml of methylene chloride, washed with saturated sodium bicarbonate and brine, dried over magnesium sulfate, and evaporated to give 146 mg of the crude product. Chromatography on silica gel eluting with 1/1 hexane-ethyl acetate gave 46 mg (37%) of the title compound as a yellow oil, which was taken directly to the next step.
Trans~ (2-Carboxy-6-hy~roxybenzoyl)benzoyloxy)-3~
hydro~cybenzami~o)perhydroazepine Trifluoroacetic Acid Salt Hydrate A solution of 46 mg (0.052 mmol) of trans-N-benzyl-4-(4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoyloxy)-3-(4-benzyloxybenzamido)azepine in 10 ml of ethanol was treated with 8.1 ~L of trifluoroacetic acid, cooled to O-C, and 22 mg of moist 10% palladium hydroxide on carbon was added. The mixture was then stirred under an atmosphere of hydrogen for 19 h at room temperature. The mixture was filtered, evaporated, and the residue was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min).
The pure fractions were pooled and evaporated and then lyophilized from water to give 9.8 mg (28%) of the title compound as a white fluffy solid. FABMS: m/z 519 (M + H).
Anal. Calc. for C28H26N208 2.5 H20 TFA; C, 53.18 H, 4.76; N, 4.13. Found: C, 53.43: H, 4.64; N, 4.30 Wo 94/20062 215 7 412 PCTtUS94tO2283 Trans 4 - t 4 - ~ 2 -Hy~roxycarbonyl - 6-hy~roxybenzoyl ) - 3, 5-dih~ oa~enzoyloxy~-3- ~4-hydlohyLe"z~ o) -1- ~phenyl~ulfonyl) -pyrrolidine ~COMPO~ND S62 ) CHzCIz H20 ~, OBn H-~FA S 02Ph C02Bn Pr~El, I OBn DMAP,~.
OBn Hæ~O~N~ EIOH. ~F ~ h OBn b~gH S02Ph BnO2Cb_OBn N
Figure AC
+ ) -Trans-3- ~ ~-benzyloxybenzamido) -~-hydroxy-l-~phenylsul f onyl ) pyrrolidine To a slurry of (+)-Trans-3-(4-benzyloxybenzamido)-4-hydroxypyrrolidine TFA (150 mg, 0.352 mmol) in H20 (8.8 ml) and CH2Cl2 (8.8 ml) were added anhydrous Na2C03 (112 mg, 3.0 eq, 1.06 mmol) then benzene sulfonyl chloride (58 ~1, 0.458 mmol, 1.3 eq), and the mixture stirred at room temperature 15 h. The solution was then diluted with CH2Cl2 (20 ml) and poured into H20 (20 ml) and methanol (4 ml). The layers were separated and the aqueous layer extracted with CH2C12 (3 x 30 ml). The organics were combined, dried (MgS0~), filtered and evaporated to a white powder (159 mg, quant yield): 1H NMR (CD30D) ~ 7.62 - (d, J = 7.7 Hz, 2H), 7.43 (d, J = 8.9 Hz, 2H), 7.35-7.30 (m, 2~57412 W094/20062 PCT~S94102283 3H), 7.25-7.10 (m, 5H), 6.80 (d, J = 8.8 Hz, 2H), 4.95 (s, 2H), 4.06-4.00 (m, lH), 3.95-3.90 (m, lH), 3.50-3.35 (m, 2H), 3.15 (dd, J = 10.6, 3.9 Hz, lH), 2.99 (dd, J = 10.8, 3.2 Hz, lH).
Trans-4-[4-~6-benzyloxy-2-~benzyloAyc~rbonyl)benzoyl]-3,5-dibenzyloAybenzoyloAy)-3- ~-benzyloxybenzamid,o)-l-(phenylsulfonyl)pyrrolidine (CONPO~nlD 639) To a solution of the previous product (159 mg, 0.352 mmol) in CH2Cl2 (6.0 ml) were added 4-dimethylaminopyridine (43 mg, 0.352 mmol, l.o eq), diisopropylethylamine (74 ~1, 0.42 mmol, 1.2 eq) then a solution of acid chloride (0.383 mmol, 1.1 eq) in CH2Cl2 (3.0 ml). The mixture was stirred at room temperature under N2 14 h. The reaction mixture was then diluted with CH2C12 (30 ml), and washed with 10% NaHCO3 (50 ml) then brine (50 ml). The aqueous layers were combined and extracted with CH2C12 (2 x 50 ml). The organics were combined, dried (MgSO~), filtered and evaporated. Flash column chromatoghraphy of the residue (2:1 hexane:ethyl acetate) on silica gel provided the title compound (183 mg, 47%): lH NMR
(CDCl3) ~ 7.77 (d, J = 6.7 Hz, 2H), 7.70 (d, J = 8.8 Hz, 2H), 7.47-7.16 (m, 14H), 7.15-7.05 (m, 6H), 7.04-6.94 (m, 3H), 6.90-6.83 (m, 3H), 6.42 (d, J = 6.7 Hz, lH), 5.30 (dt, J = 5.1, 2.6 Hz, lH), 5.18 (s, 2H), 5.13 (s, 2H), 4.78 (m, 2H), 4.76 (s, 2H), 4.72 (s, 2H), 4.64-4.60 (m, lH), 3.88 (dd, J = 12.6, 5.4 Hz, lH), 3.74-3.65 (m, lH), 3.60-3.38 (m, 2H).
Trans 4- t 4 - ~ 2 -Hydroaycarbonyl- 6-hydroAybenzoyl ) -3, 5-diL~oa~anzoyloAy]-3- (~-h2~roA~enz~i~o) -1- ~phenylsulfonyl) -pyrrolidine ~COMP0~1ND 562 ) To a solution of the previous product (183 mg, 0.164 mmol) in THF (7.4 ml) and ethanol (7.4 ml) was added Pd(OH)2 (92 mg. of a 20% by weight powder). The flask was evacuated and filled with H2 twice, then stirred under H2 (1 atm) for 20 h. The suspension was filtered through Celite, washed through with methanol (50 ml), and evaporated to a yellow oil.
Purification by HPLC (21 x 250 mm Cl8 column) provided the title compound (75 mg, 69%) as a fluffy yellow powder after ~09412W62 21~ 7 412 PCT~S94/022~
lyophilization: mp 185-208C; IR (KBr) 3402, 1709, 1636, 1608, 1232 cm ; H NMR (CD30D) ~ 7.53 (d, J = 8.2 Hz, 2H), 7.49 (d, J = 8.7 Hz, 2H), 7.32 (d, J = 7.3 Hz, lH), 7.22-7.11 (m, 3H), 7.09 (dd, J = 8.1, 7.9 Hz, lH), 6.84 (d, J = 7.2 Hz, lH), 6.61 (d, J = 8.7 Hz, 2H), 6.35 (s, 2H), 4.99 (app t, J = 2.2 Hz, lH), 4.92 (dd, J = 5.6, 2.8 Hz, lH), 3.62 (dd, J = 13.0, 4.3 Hz, lH), 3.50 (dd, J = 10.8, 6.0 Hz, lH), 3.39 (dd, J = 10.7, 2.4 Hz, lH), 3.32 (bd, J = 13.1 Hz, lH); HRMS (M + H) calcd 663.6315, found 663.1302; Anal. Calcd- for C32H26N20l2S 1 H2O:
C, 56.47; H, 4.15; N, 4.12; S, 4.71; found: C, 56.56; H, 4.17;
N, 4.09; S, 4.58.
-094/20062 2 1 5 7 1 1 2 PCT~S94/0~
Tr~ns-1-(4-hy~-o~yLehz~mi~o)-2-l4-~2-hy~roxy-6-meth benzoyl)-3,5-dihydroxybenzoyloxyl]cycloheptane ~COMPOUND 566) HO OM
O O
H~ ~ H~ , K,CO,, HMPA ~ H~
b o ~ o Figure AD
A mixture of Compound 588 (55 mg, 0.1 mmol), iodomethane (0.05 ml. 0.8 mmol), and K2CO3 (28 mg, 0.2 mmol) in HMPA (0.2 ml) was stirred at 40-C for 1.5h, and the reaction was judged incomplete by TLC. Additional iodomethane (0.025 ml, 0.4 mmol) was added and stirring was continued for 2h. at 40 C. EtOAc (15 ml) was added and the resultant mixture was washed with H2O (3 x 10 ml) and brine (10 ml), dried (MgSO4), and evaporated. The residue was purified by preparative TLC
(SiO2, multi-elution with CH2C12:5% MeOH in EtOAc = 4:1) to give a yellow solid (34 mg, 61%). IR (KBr, cm1): 1712, 1634, 1607.
FBMS: M/Z = 564 (M + 1).
W094/20062 21~ 7 ~ 12 PCT~S94/02283 Trans-~-t~-~2-Hydroxycarbonyl-6-hydroxybenzoyl)-3,5-~ihydko~L~zoyloxy]-3-(4-hyd-~yL~z~mido)-l-(5-dimethyl~min l-naphthalene sulfonyl)perhydroazepine (COMPOUND 569) O HO~
~OH OH,CH3 ~OH OH
H~o~ H ~ Cl ~I TEA ~ HHo~ Q~
,~ O N~ +~ S~ MeOH ~ ~O~N~ CH3 CF,CO2H NH CF,C02H \~ N~ SJ3~ ~ C~
569 Oo Racemic balanol (preparation described in Compound 508; 100 mg, 147 ~mol) was dissolved in methanol (1 ml) and treated with triethylamine (204 ~1, 1.47 ~mol) and dansyl chloride (39.5 mg, 146.5 ~mol) in methylene chloride (1 ml).
After stirring at room temperature for 3 h, the mixture was concentrated under vacuum to a yellow film. The residue was dissolved in DMF (2 ml) and chromatographed on a Dynamax-60 C18 column (41 mm ID x 25 cm length) using a linear gradient from 100% A (0.1% TFA and 5% acetonitrile in water) to 100% B (pure acetonitrile) over 60 m at 25 ml/min. The clean product, which eluted in 40 m, was freeze-dried to give a yellow powder (33 mg, 29%): m.p. 185-187-C dec; 1H-NMR (DMSO, 300 MHz) 1.73-2.12 (4H, m), 2.85 (6H, s), 4.30-4.41 (lH, m), 5.20 (lH, pseudo t), 6.75-6.87 (4H, m), 7.07 (lH, d, J = 8 Hz), 7.30 (2H, t), 7.39 (lH, d, J = 8 Hz), 7.62-7.70 (4H, m), 8.06 (lH, d, J
= 8 Hz), 8.24 (lH, d, J = 9 Hz), 8.50 (lH, d, J = 9 Hz), 9.91 (lH, 8), 11.68 (lH, s); IR (KBr): cm1 3399, 2361, 2340, 1702, 1677, 1635, 1607, 1542, 1507, 1462, 1425, 1371, 1319, 1283, 1237, 1200, 1139, 1103, 1064, 991, 920, 848, 793, 767, 723, 669, 581, 542, 532. Anal. Calcd. for C~oH37N3O12S 2H2O .92TFA
W094/20062 215 7 ~12 PCT~S94/022~
.15CH3CN: C, 54.37; H, 4.58; N, 4.73; S, 3.44. Found: C, 57.35; H, 4.51; N, 4.63; S, 3.17. LRMS (FAB) m/z 784.0 (783.2 calcd for C40H37N3O12S).
21~741~
wos~/20062 PCT~S94/02283 Tr~ns- ~ - t ~- ~ 2 -Carboxy-6-hydroxy~enzoyl ) -3, 5-dihydroxybenzoyloxy] -3- ~ 4 -hydroxy~enz~mido ) -1- ~ 2 -nitrobenzenesulfonyl)perhydroazepine, trifluoracetic acid salt COMPOIIND 57 3 ) o HO~=
~OH OH ~OH OH
H~o~ H~ Cl~ ~ TEA , H
~ o s ~ MeOH ~ O H~
CF,CO2H NH
CF,CO2H S~
Racemic balanol (preparation described in Compound 508; 100 mg, 147 ~mol) was dissolved in methanol (1 ml) and treated with triethylamine (204 ~1, 1.47 ~mol) and 2-nitrobenzene sulfonyl chloride (48.7 mg, 219.7 ~mol) in methylene chloride (1 ml). After stirring at room temperature for 3 h, the mixture was concentrated under vacuum to a yellow film. The residue was dissolved in DMF (2 ml) and chromatographed on a Dynamax-60 C18 column (41 mm ID x 25 cm length) using a linear gradient from 100% A (0.1% TFA and 5%
acetonitrile in water) to 100% B (pure acetonitrile) over 60 m at 25 ml/min. The clean product, which eluted in 33 m, was freeze-dried to give a yellow powder (28 mg, 26%): m.p.
158-160-C dec; lH-NMR (DMSO, 300 MHz) ~ 1.81-1.97 (3H, m), 2.06-2.17 (lH, m), 4.43-4.57 (lH, m), 5.18-5.28 (lH, m), 6.75 - (2H, s), 7.04 (lH, d, J = 8 Hz), 7.22-7.31 (3H, m), 7.36 (lH, d, J = 8 Hz), 8.05 (lH, t), 8.20 (lH, d, J = 8 Hz), 8.20 (lH, d, J = 8 Hz), 8.85 (lH, d, J = 8 Hz), 9.87 (lH, s), 11.67 (lH, s); IR (KBr): cm 3430, 3412, 1701, 1676, 1636, 1604, 1545, W094/20062 PCT~S94/02283 21~7~1~
1496, 1425, 1370, 1290, 1237, 1201, 1148, 1104, 1063, 1016, 993, 921, 874, 856, 799, 761, 738, 724, 586. Anal. Calcd. for C34H2~N30l4S 1-2 H20 1.1 TFA: C, 49.25; H, 3.71; N, 4.76; S, 3.63. Found: C, 49.22; H, 3.70; N, 4.72; S, 3.39. LRMS (FAB) m/z 735.9 (735.68 calcd for C34H29N3014S).
w094/20062 213 ~ ~ ~ 2 PCT/US94/0~
Tran8-4- ~- (2-Carboxy-6-hy~roxybenzoyl)-3,5-dihydroxybenzoyloxy]-3- (~-hydroxybenzamido) -1- ~
nitrobenzenesulfonyl)perhy~roazepine, trifluoracetic acid salt COMPO~ND 57 ~ ) o HO~
~OH OH ~=~OH OH
~~ H~ Cl` ~ TEA ~ HHo O .S ~ MeOH~ H~
~ ~ O O O ,~ j~ ,~,, NO2 CF~Co2H \~, NH CF~Co2H \ N` S
574 Oo Figure AG
Racemic balanol (preparation described in Compound 508; 100 mg, 147 ~mol) was dissolved in methanol (1 ml) and treated with triethyiamine (204 ~1, 1.47 ~mol) and 4-nitrobenzene sulfonyl chloride (48.7 mg, 219.7 ~mol) in methylene chloride (1 ml). After stirring at room temperature for 3 h, the mixture was concentrated under vacuum to a yellow film. The residue was dissolved in DMF (2 ml) and chromatographed on a Dynamax-60 C18 column (41 mm ID x 25 cm length) using a linear gradient from 100% A (0.1% TFA and 5%
acetonitrile in water) to 100% B (pure acetonitrile) over 60 m at 25 ml/min. The clean product, which eluted in 34 m, was freeze-dried to give a yellow powder (12 mg, 11%): m.p.
186-188-C dec; lH-NMR (DMSO, 300 MHz) ~ 1.80-1.97 (3H, m), 2.07-2.18 (lH, m), 3.13 (2H, pseudo t), 4.42-4.53 (lH, m), 5.17-5.28 (lH, m), 6.75 (2H, d, J = 6 Hz), 7.03 (lH, d, J = 8 Hz), 7.19 (lH, d, J = 9 Hz), 7.26 (lH, t), 7.34 (lH, d, J = 8 Hz), 7.63 (lH, d, J = 8 Hz), 7.76 (lH, d, J = 9 Hz); 7.88 (lH, d, J = 9 Hz), 8.15 (2H, d, J = 9 Hz), 8.43 (2H, d, J = 9 Hz), W094/20062 ~15 7 41 ~ PCT~S94/022~
~ - 276 -9.84 (lH, pseudo s, 11.71 (lH, pseudo s); IR (KBr): cm 3422, 3273, 3250, 3108, 3081, 2873, 2361, 2339, 1676, 1636, 1606, 1536, 1497, 1426, 1369, 1288, 1232, 1200, 1148, 1093, 1072, 1013, 960, 920, 874, 856, 761, 724, 681, 606, 568. Anal.
Calcd. for C34H29N3Ol4S 1 H2O 1.2 TFA .17 CH3CN: C, 49.17;
H, 3.67: N, 4.94; S, 3.57. Found: C, 49.16; H, 3.57; N, 4.64;
S, 3.36. LRMS (FAB) m/z 736.1 (735.68 calculated for C34HZ9N3ol4s ) -W094/20062 2 1 ~ 7 4 1 2 PCT~S94/02283 Tr~ns-3-(4-hy~roxybenz~mido)-~-t~-~2-hy~roxy-6-(2-methyl-propyloxy)c~rbonylhen~oyl)-3,5-~ihy~roxybenzoyloxy]pyrroli~ine trifluoro~cetic ~cid s~lt (COMPOUND 577) BnO~n1, (COCI)2, DMF, CH2C12 BnO~O o o~n OBn OFB~ HO", NJb~ ~, HJ~3 BOC BOC
DMAP, EtJN
H2, Pd(OH)~ / C
TFA
HO E~OH / EtOAc ~N)~ $N~ QH
BOC
TFA
~0 ~ O OH
H~
TFA
21S741`~
~ ~ - 278 -Trans-N-t-butoxycarbonyl-3-(~-benzyloxybenzam~ ~o)-~-t~-(2-benzyloxy-6-benzyloxycarbonylbenzoyl)-3,5-dibenzyloxy]-pyrrolidine 4-[4-(2-benzyloxy-6-benzyloxycarbonylbenzoyl)]-3,5-dibenzyloxybenzoic acid (1.47 mmol, 996 mg) and 15 ml anhydrous CH2Cl2 in a dry round-bottom flask were cooled in an ice/water bath under N2. To this was added oxalyl chloride (2.87 mmol, 0.25 ml) and 5 drops of DMF. This was allowed to stir for 2 hours while the bath melted. TLC (2:1 hexanes:EtOAc) indicated complete formation of the acid chloride. The solvent was removed in vacuo.
In a 200 ml dry round-bottom flask was added trans-N-t-butoxycarbonyl-3-(4-benzyloxybenzamido)-4-hydroxypyrrolidine (1.26 mmol, 500 mg) in 12 ml anhydrous CH2C12 under N2. To this was added triethylamine (3.6 mmol, 0.5 ml) and DMAP (150 mg).
A solution of the acid chloride generated above in 10 ml anhydrous CH2Cl2 was added via cannula. This was allowed to stir under N2 at room temperature overnight. The reaction mixture was then diluted with CH2C12, washed with sat. NaHCO3, brine, then dried over MgSO" and concentrated in vacuo. The crude product was purified via flash column chromatography using 5% acetone/CH2Cl2 as the eluent. Trans-N-t-butoxycarbonyl-3-(4-benzyloxybenzamido)-4-[4-(2-benzyloxy-6-benzyloxycarbonylbenzoyl) -3,5-dibenzyloxy benzoyloxy~
pyrrolidine (1.08 mmol, 1.15 g) was obtained in 8696 yield.
Trans-N-t-butoxyoarbonyl-3-(~,-hy~roxybenzamido)-~-[~-(2-hy~roxy-6-¢~,bG.~benzoyl)-3,5-dihydroxybenzoyloxy]pyrrol~dine To a 500 ml 3-neck round-bottom flask was added trans-N-t-butoxycarbonyl-3-(4-benzyloxybenzamido)-4-[4-(2-benzyloxy-6-benzyloxycarbonylbenzoyl) -3, 5-dibenzyloxy benzoyloxy]pyrrolidine (1.02 mmol, 1.08 g) in 17 ml EtOAc and 70 ml ethanol under N2. To this was added trifluoroacetic acid (2.55 mmol, 0.20 ml) and Pd(OH)2/C (730 mg) followed immediately by introduction of H2 at 1 atmosphere. After a reaction time of 3.5 hours, the reaction was flushed with N2 -~094/20062 21~ 7 ~ ~ 2 PCT/US94/02283 and filtered through Celite, rinsing with ethanol. Following concentration in vacuo, crude product (644 mg) was obtained in quantitative yield. A small portion was purified via HPLC (21 x 250 mm C18 column, gradient ~B = 0 to 50 over 60 min. where A = 0.1~ TFA, 5% CH3CN in water and B = CH3CN, 15 ml/min. W =
254 nm) for characterization and the remainder was used crude in subsequent reactions. m.p. 196C(dec). IR (KBr) 3375(br), 2978, 1704, 1660, 1637, 1607, 1506, 1426, 1368, 1231 cm . H
NMR CD30D, ~ 8.52 (d, lH), 7.72 (d, 2H), 7.49 (d, lH), 7.26 (t, lH), 7.01 (d, lH), 6.91 (s, 2H), 6.80 (d, 2H), 5.40 (m, lH), 4.63 (m, lH), 3.87 (m, 2H), 3.50 (m, 2H), 1.47 (s, 9H). LRMS
(M + 1) cacld for C31H31N2O12 623.2, found 623.2. Anal. Calcd for C3lH3lN2O12 1-5 H2O: C, 57.317; H, 5.120; N, 4.312. Found: C, 57.26; H, 5.18; N, 4.47.
Trsns-N-t-butoxyc~rbonyl-3-(~-hydroxybenz~mi~o)-~-t4-~2-hydroxy-6-~2-methylpropyloxy)¢~rbonylbenzoyl)-3,5-dihydroxybenzoyloxy]pyrrolid~ne (COMPOUND 6~6) To a round-bottom flask was added trans-N-t-butoxycarbonyl-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-carboxylbenzoyl)-3,5-dihydroxybenzoyloxy]pyrrolidine (0.lS
mmol, 95 mg) in 5 ml DMF. To this was added NaHCO3 (0.23 mmol, 19 mg) and 1-iodo-2-methylpropane (0.75 mmol, 0.09 ml). This was allowed to stir under N2 for 7 days with additional 1-iodo-2-methylpropane being added each day. A total of 48 additional equivalents (7.2 mmol, 0.83 ml) were added over the reaction period. The reaction mixture was diluted with EtOAc and washed with water 3 times. The aqueous layer was back extracted with EtOAc and the organic layers combined and dried over MgSO4 then concentrated in vacuo. The crude product was purified via HPLC
(21 x 250 mm C18 column, gradient %B = 25 to 100 over 60 min.
where A = 0.1% TFA, and 5% CH3CN in water, B = CH3CN, 15 ml/min.
W = 254 nm) to isolate trans-N-t-butoxycarbonyl-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-(2-methylpropyloxy)-carbonylbenzoyl)-3,5-dihydroxybenzoyloxy]pyrrolidine (46.5mg, 44% yield).
WO 9~/20062 215 7 4 ~ 2 PCT/US94/02283 Trans-3- ~4-hydroxybenzami~o) -4- t4- ~2-hydroxy-6- ~2-methylpropyloxy)carbonylbenzoyl)-3,5-dihy~roxybenzoyloxy]-pyrrolidine trifluoroacetic acid ~alt ~COMPOlrND 577) Trans-N-t-butoxycarbonyl-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6- (methylpropyloxy) carbonylbenzoyl) -3, 5-dihydroxybenzoyloxy]pyrrolidine (46.5 mg, 0.068 mmol) was dissolved in 0.75 ml neat trifluoroacetic acid and allowed to stir at room temperature under N2 for 45 minutes at which time TLC (75% CH2C12, 24% MeOH, 1% (10% aq.) NH40H) indicated the reaction was complete. This was diluted with toluene and concentrated in vacuo to yield 42.7 mg (91% yield) of crude product. Purification via HPLC (21 x 250 mm C18 column, gradient %B = 0 to 100 over 60 min. where A = 0.1% TFA, 5%
CH3CN in water, B = CH3CN, 15 ml/min. W = 254 nm) yielded trans-3- (4-hydroxybenzamido) -4- [4- (2-hydroxy-6- (2-methylpropyloxy ) -carbony1benz oy1 ) - 3, 5 -dihydroxybenzoyloxy~pyrrolidine trifluoroacetic acid salt (Compound 577, 32 mg, 6896 yield) as a yellow solid. m.p.
150-156-C (dec.). IR (KBr) 3191 (br), 2968, 1677, 1607, 1508, 1426, 1373, 1293, 1201 cm-l. lH NMR, DMSO-d6, ~, 11.74 (s, 2H), 10.13 (s, lH), 10.03 (s, lH), 9.20 (br, NH), 8.51 (d, lH), 7.74 (d, 2H), 7.44 (d, lH), 7.35 (t, lH), 7.12 (d, lH), 6.95 (s, 2H), 6.84 (d, 2H), 5.50 (m, lH), 4.60 (m, lH), 3.88 (d, 2H), 3.72 (m, 2H), 3.52 (m, 2H), 1.80 (q, lH), 0.83 (d, 6H).
LRMS (M + 1) calcd for C30H3lN2Olo 579.20, found 579.1. Anal.
calcd for C30H30N2Olo C2HF3O2 1.3 H2O: C, 53-68; H~ 4-73; N~
3.91. Found: C, 53.78; H, 4.54; N, 3.99.
~o9~l2oo62 ~1574~2 Tr~n8~ -hy~roxybenz~mi~o) -2- t~- (2-hy~roxy-6-hy~roxyc~rbonylbenzoyl)-3,5-~ihy~roxybenzoyloxyl]cyclohept~ne (COMPO~ND 588) C~ N-~CO~ CH~ (9 I~OH ~O (~ N~ TBDI~tS Cl OBn OTB0MS H?~Pd-C OTBDMS . ' H ~) N~ M OH (~ BnO~ CO~H, THf TBDMS~N~
CO~Bn OBn OlloO Olln g~n O n Bu,NF THF H~j o EI,N,DMAP,CH2CI, BnO O N~
~J ~ O
a~,H
H?~Pd(OH)~ C ~H ~ H
M~OH THF o ~
21~7~12 ~ ; - 282 -NaOAc (426 mg, 5.2 mmol) was dissolved in a solution of peroxyacetic acid in acetic acid (32 wt. %, 21.9 ml, 104 mmol) and the resultant solution was added dropwise over 30 min. to a mixture of cycloheptene (10 g, 104 mmol) and Na2CO3 (44.1 g, 416 mmol) in CH2C12 (100 ml) at 5 C. The mixture was allowed to stir at room temperature for 3h with occasional cooling using a water bath. All solid material was removed by filtration, and the filtrate was distilled at atmospheric pressure using a vigreux column to remove most of the CH2Cl2, giving a colorless liquid which was shown by Hl NMR to be a mixture of the epoxide and CH2C12. The purity was estimated to be ca. 73% in the epoxide (total 15.93 g, 100% crude yield).
This material was used without further purification in the next step.
A mixture of the epoxide from the previous step (10 g, 65 mmol), NaN3 (27.5 g, 423 mmol), NH4Cl (10.44 g, 195 mmol), MeOH (162 ml), and H2O (20 ml) was stirred at reflux for 24h.
The solid was removed by filtration and the filtrate was evaporated. The residue was treated with 0.5 N aq. NaOH (50 ml) and extracted with CH2C12 (3 x 30 ml). The combined CH2Cl2 extracts were washed with H2O (50 ml), brine (50 ml), dried (MgSO4), and evaporated to give the azide (8.39 g, 83%).
The azide (7 g, 45 mmol) in N,N-dimethylformamide (10 ml) was added to a solution of t-butyldimethylsilylchloride (6.8 g, 45 mmol) and imidazole (3.07 g, 45 mmol) in N,N-dimethylformamide (35 ml). The resultant mixture was stirred at room temperature for 16h, diluted with Et2O (70 ml), washed with H2O (4 x 30 ml) and brine (50 ml), dried (MgSO4), and evaporated to give a pale yellow oil (11.33 g, 93%).
A mixture of the product of the previous reaction (8.1 g, 29.95 mmol) and 5% Pd on carbon (1.59 g, 2.5 mol. %) in MeOH
was stirred vigorously under 1 atm H2 at room temperature for 20h. The catalyst was removed by filtration over Celite and the filtrate was evaporated. The residue was purified by flash chromatography (SiO2, Et2O: Hexane = 2:1) to give the amine as a colorless oil (5.27 g, 72~).
~094/20062 213 7 ~1~ PCT/US94/02283 A mixture of 4-benzyloxybenzoic acid (3.287 gj 14.40 mmol) and 1,1'-carbonyldiimidazole (2.335 g, 14.40 mmol) in THF
was stirred at room temperature for 2h. To the resultant slurry was added a solution of the amine product of the previous reaction (3.07 g, 12.56 mmol) in THF and stirring was continued at room temperature for 24h. The mixture was diluted with CH2Cl2 (40 ml), washed with H20 (3 x 15 ml) and brine (2 x 15 ml), dried (MgS04), and evaporated. The residue was purified by flash chromatography (SiO2, Et20 : hexane = 1:5) to give the amide as a white powder (4.69 g, 82%).
Tetrabutylammonium fluoride in THF (lM, 10 ml, 10 mmol) was added to a solution of the amide product from the previous reaction (3.8 g, 8.36 mmol) in THF and the resultant yellow solution was stirred at room temperature for 2h. The mixture was poured into CH2C12 (150 ml), washed with H20 (3 x 30 ml) and brine (2 x 30 ml), dried (MgS04), and concentrated to about 30 ml. The precipitate was collected by filtration and washed with CH2Cl2 (3 x 3 ml). The combined filtrate and washes were concentrated to about 10 ml, and the resultant precipitate was collected, washed with CH2C12 (2 x 2 ml), combined with the first crop and dried under vacuum. This gave a white solid (2.24 g, 79%). The mother liquors were combined and chromatographed (SiO2, Et20 : hexane = 1:5 followed by CH2Cl2 to give an additional 256 mg of the product, total yield: 89%.
To a stirred solution of 4-(2-benzyloxycarbonyl-6-benzyloxy)benzoyl-3,5-dibenzyloxybenzoylchloride (350 mg, 0.52 mmol) in CH2Cl2 (1.5 ml) was added oxalyl chloride (0.067 ml, 0.77 mmol) and one drop of N,N-dimethylformamide. The resultant solution was stirred at room temperature for 2h and then evaporated to dryness. The residue was dissolved in CH2Cl2 (1 ml) and added to a mixture of the product of the preceding reaction (177 mg, 0.52 mmol) and Et3N (105 mg, 1.04 mmol) in CH2Cl2 (2 ml). The mixture was stirred at room temperature for 17h, diluted with CH2Cl2 (15 ml), washed with H20 (3 x 10 ml), dried (MgS04), and evaporated. The residue was chromatographed wos4/20062 215 7 ~12 PCT~S94/02283 (sio2~ Et2O: hexane = 1:1, followed by Et2O: hexane: CH2Cl2 =
1:1:1) to give a white solid (166 mg, 32%).
Pd(OH)2 on carbon (20 wt. % contains > 50% moisture;
22 mg, 0.016 mmol) and MeOH (1.6 ml) were added to a solution of the product of the prec~ing reaction (160 mg, 0.16 mmol) in THF (1.6 ml) and the resultant mixture was stirred under 1 atm H2 at room temperature for 17h. The catalyst was removed by filtration over Celite and the Celite pad was washed with MeOH
(15 ml). The filtrate and washes were combined and evaporated to give a yellow solid (83 mg, 95%) (COMPOUND 588). m.p. 186-C
(dec). Anal. calcd for C29H2,NOlo-lH2O: C, 61.37; H, 5.15; N, 2.47. Found: C, 61.41; H, 5.29; N, 2.36.
2 1 ~ 741~
Trans-3- (~-hy~lohyLt:nz~lmido) -4- t4- ~2-hy~roxy-6-hy~ro2ycarbonyl benzoyl) -3,5-~ihy~roxybenzoyloxyl]pyrroli0inium trifluoroacetate (COMPO~ND 589) ~nd 1-}~exa~ecanoyl-trzms-3-~hy~GhyL ~nzami~o) -~.- t 40 ~2-hy~roxy-6-hy~roxycarbonylbenzoyl ) -3, 5-~ihy0roxybenzoyloxyl ] pyrrol i~ine ~ COMPOIJND S 9 0 ) co2~
e~~i C~ ~n ~H~en EloN~ DMAP, CH,C~I
o~L OEln O 06n CO~H
~ ~ c~(c~"coa MeOH, THF O ~ O
CF,CO2H
C~2H
~,~OH D~ H
HO~ ~H~J
,~0,~ 0 0~ (CH~"C~
To a stirred solution of 4(2-benzyloxycarbonyl-6-benzyloxy)benzyl-3,5-dibenzyloxybenzoylchloride (350 mg, 0.52 mmol) in CH2Cl2 (1.5 ml) was added oxalyl chloride (0.067 ml, ~og~/20062 215 7 412 PCT/US94/02283 0.77 mmol) and one drop of N,N-dimethylformamide. The resultant solution was stirred at room temperature for 2h and then evaporated to dryness. The residue was dissolved in CH2Cl2 tl.5 ml) and added to a-mixture of trans-3-(4-hydroxy-1-benzyloxycarbonyl pyrrolidine (232 mg, 0.53 mmol; for preparation see Compound 585), Et3N (0.14 ml, 1.04 mmol), and 4-(N,N-dimethylamino)pyridine (6 mg, 0.052 mmol) in CH2Cl2 (1.5 ml). The mixture was stirred at room temperature for 17h, diluted with CH2Cl2 (15 ml), washed with H2O (3 x 10 ml), dried (MgSO4), and evaporated. The residue was chromatographed (SiO2, Et2O:hexane = 1:1, then EtzO:hexane:CH2Cl2 = 1:1:0.5) to give the ester (230 mg, 40%) (COMPOUND 652).
To a solution of Compound 652 (200 mg, 0.18 mmol) in THF (1.8 ml) were added MeOH (1.8 ml), Pd(OH)2 on carbon (20 wt.~, contains 2 50% moist; 25 mg, 0.018 mmol), and CF3CO2H (41 mg, 0.36 mmol). The mixture was stirred at room temperature under 1 atm H2 for 17h. The volatile components were removed by evaporation, and the residue was taken up in MeOH, filtered through Celite, and evaporated to give a yellow solid (88 mg, 72~) (COMPOUND 589).
Palmitoyl chloride (10 mg, 0.038 mmol) was added to a stirred solution of Compound 589 (24 mg, 0.038 mmol) in pyridine (0.4 ml). The mixture was stirred at room temperature for 16h and TLC showed that the reaction was incomplete. More palmitoyl chloride (5 mg) was added and stirring was continued for 16h. The reaction mixture was evaporated to remove pyridine leaving a yellow syrup with some solid material which was shown to contain starting material by lH NMR. This material was dissolved in pyridine (0.4 ml) and treated with palmitoyl chloride (15 mg). The mixture was stirred at room temperature for 24h, evaporated, and chromatographed (SiO2, EtOAc followed by S% HOAc in acetone) to give a yellow solid (19 mg, 66%) which was shown by lH NMR to be the desired amide (Compound 590) with some contamination. m.p. 175-178C (dec).
Anal. calcd for C42H52N2Oll-3H2O: C, 61.90; H, 7.17; N, 3.44.
Found: C, 61.85; H, 7.07; N, 3.62.
_~094/20062 - 287 - PCT~S94/02283 Tran~-3-~-hydL~L~z~mido)-4-[4-(2-hydroxy-n~phth-1-oyl)-3,5-dihydroxybenzyoyloxyl]pyrrolidine trifluoroacetic ~ci~ s~lt (COMPOUND 591) H~;;5 E~,N. O~P, C~
0~ OE~n OJ` Oan ~H 01 1 U O~, THF
N CF~
To a mixture of 4-(2-benzyloxy-naphth-1-oyl)-3,5-dibenzyloxylbenzoic acid (297 mg, 0.5 mmol) and 2 drops of N,N-dimethylformamide in CH2C12 (1.5 ml) was added oxalyl chloride (0.065 ml, 0.75 mmol) dropwise over 2 min. The resultant mixture was stirred at room temperature for 2h and the volatile components were evaporated. The remaining yellow solid was dried under vacuum for lh, dissolved in CH2Cl2 (1.5 ml), and added to a solution of trans-3-(4-hydroxybenzamido-4-hydroxy-1-benzyloxycarbonyl pyrrolidine (168 mg, 0.38 mmol; for preparation see Compound 585), Et3N (0.1 ml, 0.76 mmol), and 4-(N,N-dimethylamino)pyridine (5 mg, 0.038 mmol) in CH2C12 at 5C.
wos4noo62 215 ~ 4 ~- ~ PCT/US94/022~
The mixture was stirred at room temperature for 16h, diluted with CH2Cl2 (10 ml), washed with H2O (3 x 10 ml), dried (MgSO4), and evaporated. The residue was chromatographed (SiO2, Et2O:hexane = 1:1, followed by Et2O:hexane:CH2Cl2 = 1:1:0.5) to give a white solid (336 mg, 66%) (COMPOUND 653).
Pd(OH)2 on carbon (20 wt.%, contains > 50% moist; 21 mg, 0.015 mmol), followed by CF3CO2H (34 mg, 0.3 mmol) and MeOH
(1.5 ml), was added to a solution of Compound 653 (155 mg, 0.15 mmol) in THF (1.5 ml). The resultant mixture was stirred at room temperature under 1 atm H2 for 2Oh. The catalyst was removed by filtration over Celite and the Celite pad was washed with MeOH (20 ml). The combined filtrate and washes were evaporated to give a yellow solid (85 mg, 88%) (COMPOUND 591).
~ ~O9~/20062 2157~12 l-Isopropyl-tr~n~-3-(~-hydroxybenz~mi~o~ y~2~by ~ ~y~
hydroxyc~rbonylhe~7cyl)-3~5-dihy~roxybenzoyloxyl]pyrrolidinium trifluoro~cet~te (COMPOUND 592) OBn O~n ,~ ~,O~;
~S ~, ~ o ~=/ NBH~. ~H)--\ O E~,~. DMAP, CHICI~
H~, H CF,CO,H
~eO ~e o ~H~ H, Pd(OH~,-C ~o c~ TF~ 0~. T\~F O ~ o ~ Cf~
6~ 592 W09~i20062 215 7 ~ PCT/US94/02283 To a solution of trans-3-(4-benzyloxybenzamido)-3-hydroxy pyrrolidine (213 mg, 0.5 mmol) in acetic acid (2.5 ml) was added NaBH4 (95 mg, 2.5 mmol) in small portions. After the H2 evolution had ceased (ca. 10 min.) acetone (0.18 ml, 2.5 mmol) was added and the mixture was stirred at room temperature for 16h. The reaction mixture was basified with 2N KOH and the resultant cloudy mixture was extracted with CH2C12 (3 x 15 ml).
The combined CHzCl2 extracts were dried (MgSO4), and evaporated to give a colorless oil (163 mg, 92%).
Oxalyl chloride in CH2C12 (2 M, 0.39 ml, 0.78 mmol) was added dropwise to a solution of 4-(2-benzyloxy-6-benzyloxycarbonylbenzoyl)-3,5-dibenzyloxybenzoic acid (353 mg, 0.52 mmol) and a drop of DMF in CH2C12 (2 ml) at 5 C. The mixture was stirred at room temperature for 2h, then evaporated to remove the solvent and excess oxalyl chloride. The residue was dried in vacuo for lh, dissolved in CH2C12 (1 ml), and added to a mixture of the product of the preceding reaction (142 mg, 0.4 mmol), Et3N (81 mg, 0.8 mmol), and DMAP (6 mg, 0.054 mmol) in CH2Cl2 (2 ml) at 5-C. The mixture was stirred at room temperature for 17h, diluted with CH2C12 (15 ml), washed with H2O (3 x 10 ml), dried (MgSO4), and evaporated. The residue was chromatographed (SiO2, Et2O:CH2Cl2:hexane = 1:1:1 followed by Et2O:CH2Cl2:hexane = 2:2:1) to give a pale yellow oil (82 mg, 20%) (COMPOUND 654).
Pd(OH)2 on carbon(20 wt%, contains <50% moist, 11 mg, 0.008 mmol), trifluoroacetic acid (18 mg, 0.16 mmol), and MeOH
(1 ml) was added to a solution of Compound 654 (84 mg, 0.08 mmol) in EtOAc (1 ml) and the mixture was stirred under 1 atm H2 contained in a balloon at room temperature for 16h. The mixture was filtered through Celite and the Celite pad was washed with MeOH (5 ml). The combined filtrates were evaporated to give a yellow solid (32 mg, 59%) (COMPOUND 592).
IR (KBr, cm ):1705, 1676, 1636, 1607. FBMS: M/Z = 565 (M + 1).
2157~12 ~O g~/20062 - 291 - PCT/US94/02283 Trans-~- t~- (2-carboxy-6-hydroxybenzoyl) -3,5-~ihy~roxybenzoyloxy]-3- (4-hy~roxybenzami~o) -1-[ (phenylamino) carbonyl] -pyrroli~ine (COMPO~JND 593) NE~ OBn H-TFA O~N
H
C 02Bn iPr2NE~, ¦ OBn DMAP, ~3 c OC 1 OBn 08n H~OhN~ Pd(OHk ~ O OBn H02C~ooH N 2 ~-08n I Fh (~) -Trans-~-Hydroxy-3- (~-benzyloxybenz~mido)-l-tphenylamino)carbonyl]-pyrrolidine To a solution of (+) trans-4-hydroxy-3-(4-benzyloxybenzamido pyrrolidine TFA (100 mg, 0.235 mmol) in methanol (4.7 ml) were added triethyl amine (66 ~L, 0.470 mmol, 2.0 eq) then phenyl isocyanate (26 ~L, 0.235 mmol, 1.0 eq).
The cloudy mixture was stirred at room temperature under N2 45 min, then poured into H2O (30 ml) and extracted with CH2Cl2 (3 x 25 ml). The organic layers were combined, dried (MgSO4), filtered and evaporated to a white solid (82 mg, 81%): 1H NMR
(300 MNz, CD30D ~ 7.60 (d, J = 8.8 Hz, 2H), 7.00-7.25 (m, llH), 6.85 (d, J = 8.9 Hz, 2H), 6.79 (d, J = 7.1 Hz, lH), 4.93 (s, 2H), 4.20-4.25 (m, lH), 4.16 (dd, J = 8.2, 3.1 Hz, lH), 3.73 W094/20062 21 S 7 ~12 ~ ~ PCT~S94/02283 (dd, J c 11, 6.4 Hz, lH), 3.57 (dd, J = 11.4, 5.1 Hz, lH), 3.12 (dd, J = 11.0, 3.5 ~z, lH), 3.26 (dd, J = 11.1, 2.8 Hz, lH).
( + ) -Tran~-~- t ~ - ~ 6-benzyloxy-2- ( benzyloxycarbonyl ) benzoyl ] - 3, 5-~ibenzyloxybenzoyloxy) -3- (~.-benzyloxybenzamido) -1-(phenylamino) c~rbonyl]-pyrrolidine (CON~uNv 655) To a solution of the product of the preceding reaction (80 mg, 0.19 mmol), 4-dimethylaminopyridine (26 mg, 0.21 mmol, 1.1 eq) and diisopropylethylamine (37 ~L, 0.21 mmol, 1.1 eq) in CH2Cl2 (3.2 ml) was added a solution of acid chloride (0.21 mmol) in CH2C12 (1.6 ml). The mixture was stirred at room temperature under N2 18 h, then poured into 5% HCl (30 ml) and extracted with CH2Cl2 (3 x 25 ml). The organic layers were combined, dried (MgSO4) filtered and evaporated. Flash column chromatography of the golden residue (1.1 hexanes:ethyl acetate) on silica gel provided the title product (110 mg, 53%) as an off white foam: 1H NMR (300 MHz, CDCl3) ~ 7.89 (d, J = 8.8 Hz, 2H), 6.9-7.6 (m, 26H), 6.82 (d, J = 7.5 Hz, 2H), 5.4-5.5 (m, lH), 5.12 (s, 2H), 5.02 (s, 2H), 4.80 !m, lH), 4.76 (s, 4H), 4.68 (s, 2H), 4.0-4.05 (m, lH), 3.95-4.0 (m, lH), 3.65-3.7 (m, lH), 3.6-3.65 (m, lH).
Tr~ns- ~ - t 4 - ~ 2 -C~rboxy- 6-hydroxybenzoyl ) -3, 5-dihydroxybenzoyloxy] -3- (4-hy~roxybenzami~o) -~-~phenylamino) carbonyl]-pyrrolidine ~COMr~u~v 593) To a round bottom flask containing Compound 655 (110 mg, 0.101 mmol) were added Pd(OH)2 (22 mg of a 20% powder) then THF (2.2 ml) and ethanol (2.2 ml). The flask was evacuated and filled with H2 twice, then allowed to stir under H2 (1 atm) for 23 h. The suspension was filtered, washed through with methanol (40 ml) and evaporated to a yellow solid.
Purification by reverse phase HPLC (618 column) provided the title product as a yellow powder after lyophilization (24.9 mg, 39%): m.p. 183-208 (dec); IR (KBr) 3458, 3336, 1719, 1679, 1622, 1227, 760 cm ; lH NMR (300 MHz, CD30D) ~ 7.53 (d, J = 8.6 Hz, 2H), 7.28 (d, J = 7.7 Hz, lH), 7.20 (d, J = 7,7 Hz, 2H), 7.0-7.1 (m, 3H), 6.82 (d, J = 8.3 Hz, 2H), 6.73 (s, 2H), 6.62 215741~ ~
(8.7, J = d Hz, 2H), 5.25-5.35 (m, lH), 4.5-4.6 (m, lH), 3.75-3.9 (m, 2H), 3.4-3.5 (m, 2H): MS m/e calc'd for C33H2~N3Oll:
642.1732, found 642.1865; Analysis calc'd for C33H27N3O1l-0.5 TFA-1.25 H2O: C,56.63; H, 4.19; N, 5.83; found: C, 56.85; H, ~4.26; N, 5.96.
W094l20062 ~lS 7 4 1 2 - 294 - PCT~S94/02283 ~)Trans-4t4-(2-Carboxy-6-hy~roxybenzoyl)-3,5-dihydroxybenzoyloxy]-3-(4-hydroxybenzamido)-1-t(methylamino)c~rbonyl]-pyrroli~ine (COMPOUND S9~) HO~p~ , NE~3 ~ ~3`08n H-lFA H
C02Bn Pr2NEt, ¦ OB n DMAP, ~'COCI
OBn OBn H 0~0 ~ EdlOHHkHF _~
~o h OH Oq~ ~j OBn H02C~b~ o~N,CH3 BnO2C~_gBn N
(~)-Trans-4-Hydroxy-3-(~-benzyloxybenzamido)-1-t(methylsmino)c~rbonyl]-pyrrolidine To a solution of (+)-trans-4-hydroxy-3-(4-benzyloxybenzamido)pyrrolidine TFA (100 mg, 0.235 mmol) in methanol (4.7 ml) were added triethyl amine (66 ~1, 0.470 mmol, 2.0 eq) then methyl isocyanate (14 ~L, 0.235 mmol, 1.0 eq). The cloudy mixture was stirred at room temperature under N2 1.5 h, then more methyl isocyanate (14 ~L, 1.0 eq) was added. The mixture was allowed to stir 10 min more, then poured into H20 (30 ml) and extracted with CH2C12 (3 x 25 ml). The organic layers were combined, dried (MgS04) filtered and evaporated to ~ W094/20062 21~ 7 412 PCT/US94/02283 a white solid (83 mg, 95%): H NMR (300 MHz, CD30D) ~ 7.60 (d, J = 8.8 Hz, 2H), 7.1-7.25 (m, 5M), 6.85 (d, J = 8.8 Hz, 2H), 4.66 (s, 2H), 4.15 (dd, J = 6.5, 3.4 Hz, lH), 4.10 (dd, J =
5.0, 3.4 Hz, lH), 3.57 (dd, J = 10.8, 6.5 Hz, lH), 3.43 (dd, J
= 11.1, 5.2 Hz, lH), 3.1-3.2 (m, 2H), 2.53 (s, 3H).
(+)-Trans-~-t~-~6-benzyloxy-2-~benzyloxycarbonyl)benzoyl]-3,5-dibenzyloxybenzoyloxy) -3- ~-benzyloxybenzamido) -1-[ ~methylamino) c~rhonyl~ -pyrrolidine ~COMPOUND CS6) To a solution of the product of the prececeding reaction (85 mg, 0.23 mmol), 4-dimethylaminopyridine (28 mg, 0.23 mmol, 1.1 eq) and diisopropylethylamine (40 ~L, 0.23 mmol, 1.1 eq) in CH2C12 (3.9 ml) was added a solution of acid chloride (0.25 mmol) in CH2C12 (1.9 ml). The mixture was stirred at room temperature under N2 16 h, then poured into 5% HCl (30 ml) and extracted with CH2C12 (3 x 25 ml). The organic layers were combined, dried (MgS04) filtered and evaporated. Flash column chromatography of the golden residue (98:2 CH2Cl2:methanol) on silica gel provided the title product (159 mg, 67%) as an off white solid: lH NMR (300 MHz, CDCl3) ~ 7.84 (d, J = 8.7 Hz, 2H), 7.3-7.4 (m, 5H), 7.1-7.3 (m, 16H), 7.06-7.08 (m, 6H), 6.9-7.0 (m, 3H), 6.84 (d, J = 7.6 Hz, 2H), 5.49 (m, lH), 5.14 (s, 2H), 5.07 (s, 2H), 4.78 (s, 4H), 4.70 (m, lH), 4.70 (s, 2H), 4.40 (m, lH), 3.95-4.0 (m, 2H), 3.53-3.60 (m, 2H), 2.80 (d, J = 4.5 Hz, 3H).
~ + ) -Trans- ~ - t ~ - ~ 2 -Carboxy- 6 -hydroxybenzoyl ) - 3, 5 -dihy~ro~cyenzoyloxy] -3- ~.-hy~roxybenzamido) -1-~methyl~ino) carbonyl]-pyrrolidine ~COMPOIJND 594) To a round bottom flask containing Compound 656 (159 mg, 0.154 mmol) were added Pd(OH)2 (40 mg of a 20% powder) then THF (3.0 ml) and ethanol (3.0 ml). The flask was evacuated and filled with H2 twice, then allowed to stir under H2 (1 atm) for 21 h. The suspension was filtered, washed through with methanol (50 ml) and evaporated to a yellow solid.
Purification by reverse phase HPLC (C18 column) provided the title product as a yellow powder after lyophilization (68.7 mg, ~VO 94/20062 21~ 7 412 PCT/US94/02283 77%): m.p. 178-198 (dec); IR (KBr) 3385, 1714, 1605, 1236, 763 cm; H NMR (300 MHz, CD30D) ~ 7.53 (d, J = 8.7 Hz, 2H), 7.30 (d, J = 7,7 Hz, lH), 7.06 (dd, J = 8.1, 7.9 Hz, lH), 6.80 (d, J = 8.3 Hz, lH), 6.70 (s, 2H), 6.62 (d, J = 8.7 Hz, 2H), 5.2-5.3 (m, lH), 4.4-4.5 (m, lH), 3.6-3.8(6 line mult, 2H), 3.2-3.4 (m, 2H), 2.53 (s, 3H); MS m/e calc'd for C28H25N3011: 580-1567~
found 580.1481; Analysis calc'd for C2~H25N3O11-0.3TFA 0.8H20:
C, 54.32; H, 3.96; N, 6.72; found: C, 54.61; H, 4.32; N, 6.68.
~O9~/20062 21 r 7 ~
- 297 - ~ PCT~S94/02283 ~ tr~ns-3-~3,~-~ihy~roxybenz~mido)-~-t~-(2-c~rboxy-6-hy~roxybenzoyl)-3,5-dihy~roxy]benzoyloxypyrroli~ine trifluoro~cetic ~cid ~lt (COMPOUND 675) BnO
-- ~ O Jonesox.
BnO ~ ~
~-J H 93%
OBn H NH2 ~ OBn - BnO H _ + Bn ~ O NaOH,CH2C12 HO\ NL_ CB OH 99% N
Z B
z BnO
O 1.(COCI)2cat.DMF,~ OBn OBn OBnCH2CI2 ~ ~
2. Et3N, DMAP,BnLOnO ~ H ~ OBn B~sOno~~OH CH2CI2.70%
O N
/ B
HO / 2. Pd(H)2 ,~--o / EtOAc-EtOH, TFA
OH OH
H~ ~ H ~ OH
O ~ N ~
WO9~/20062 21~ 7 ~1 2 PCT/US94102t83 Jones reagent (6ml) was added to a solution of 3,4-dibenzyloxybenzaldehyde (Aldrich, 1.0 g, 3.14 mmol) in acetone (20 ml) until the reaction remained the color of Jones reagent. The excess of Jones reagent was destroyed by adding i-PrOH and acetone was removed in vacuo. The slurry residue was taken into EtOAc, washed with brine, dried with Na2SO4, and concentrated to afford off-white solids (0.98 g, 93~).
To a suspension of 3,4-dibenzyloxybenzoic acid (0.467 mg, 1.40 mmol) in anhydrous CH2C12 (5 ml) was added cat. DMF and oxalyl chloride (2.0 M solution in CH2C12, 1.92 ml, 3.85 mmol) at room temperature The mixture was kept for stirring at room temperature for 2 hr. Solvents were removed and the acid chloride residue was taken into CH2C12 (5 ml) after drying over the vacuum for lhr. To a biphasic reaction mixture of N-CBZ-3-amino-4-hydroxypyrrolidine (0.74 M solution in CH2Cl2, 300 mg, 1.7 ml, 1.27 mmol) in CH2C12 (10 ml) and 1 N NaOH (13.0 ml) was added a solution of 3-benzyloxybenzoic acid chloride in anhydrous CH2C12 (5 ml). The resulting mixture was vigorously stirred at room temperature for 3h. The reaction mixture was diluted with EtOAc, washed with brine, and dried over Na2SO4.
The crude product after concentration was triturated in Et2O
and EtOAc to afford white solids (quantitative yield).
To a solution of benzophenone acid (265 mg, 0.39 mmol) in CH2Cl2 (5 ml) was added cat. DMF and oxalyl chloride (2.0 M
solution in CH2C12, 0.488 ml, 0.976 mmol) at room temperature The mixture was kept for stirring at room temperature for 2 hr.
Solvents were removed and the acid chloride residue was taken into CH2Cl2 (5 ml) after drying over the vacuum for lhr.
A solution of amidoalcohol (215.5 mg, 0.39 mmol), Et3N
(197.3 mg, 272 ~L, 1.95 mmol) and DMAP (47.6 mg, 0.39 mmol) in CH2Cl2 (5 ml) was treated with the freshly made acid chloride-CH2Cl2 solution (5 ml) at 5C. The reaction mixture was allowed to stir at room temperature for 3h and then chromatographed on silica gel eluting with 2:3 to l:l/EtOAc:Hexane. The product was obtained as fluffy white solids (332 mg, 70%).
~O 94/20062 215 ~ PCT/US94/02283 The product of the previous reaction (320 mg, 0.264 mmol) was dissolved in EtOAc-HOEt (1:1, 25 ml) and treated with TFA (cat.) followed by 10% Pd(OH)2 (170 mg, 60 mol%). The mixture was subject to hydrogenolysis at 50 psi for 20hr.
Solvents were concentrated after filtering through a pad of celite, and the residue was dissolved in DMF (1.0 ml) and loaded onto HPLC; conditions: A-0.1% TFA 5%CH3CN/H2O, B-100%
CH3CN, 0-50% B over 60 min, 25 ml/min, 41 x 300 mm C18 column.
Fractions (one/min) 37-40 were combined, partially concentrated, and lyophilized to afford fluffy yellow solids (113, 65%). (COMPOUND 675) m.p. 210-213 (dec)-C. IR (KBr) cm 1 3391, 3246, 1717, 1676, 1636, and 1603. Anal. Calcd. for C26H22N2O1l-2-0H2O-1.0C2HF3O2: C, 48.84; H, 3.95; N, 4.07. Found:
C, 48.75; H, 3.63; N, 4.07. LRFAB (M + 1): 579.
wosl/20062 21~ 7 ~ 12 PCT/US94/02283 (+)-Trans-2-t4-(6-hydroxy-2-(carboxyl)benzoyl)-3,5-dihydroxybenzoyloxy]-1-(2-hydroxybenzamido)cyclopentane ~COMPOUND 681) 2 H2,PdonC HO
3.2NKOH,THF h CIOC~3 ~ BnO
BnO
~0 OBn COCI
CH2CI2, DIPEA, DMAP
BnO
Ho2c~ N)~[3 H2 ~0b~1HlB~3 OH OBn FIGURE AP
~2-Benzyloxybenzoyl)chloride To a solution of 2-benzyloxybenzoic acid (684 mg, 3.00 mmol) in CH2Cl2 (15 ml) were added dimethylformamide (1 drop) then oxalyl chloride (3.0 ml of a 2.0 M solution in CH2C12, 6.00 mmol, 2.0 eq). The solution was stirred at room temperature 1 h, then evaporated and the light yellow semi-solid was placed on the vacuum pump before use.
DEMANDES OU BREVETS VOLUMINEUX
LA PRÉSENTE PARTIE DE Ctl I E DENIANDE OU CE BREVET
COMPREND PLUS D'UN TOME. - -CECI EST LE TOME I DE
NOTE: Pour les tomes additionels, veuillez contacter le Bureau canadien desbrevets JU~ABO APPLICATIONS/PATENTS
THIS SECTION OF THE APPI ICATION/PATENT CONTAINS MORE
THAN ONE VOLUME
THIS IS VOLUME L_ OF ~
NOTE: For additional vclumes please c~ntact the Canadian Patent Office
~.-R~-~- [~3-hy~roxycarbonyl)-2-pyridinyl)c~rbonyl)-3,5-dihydro~cybenzoylo~cy]-3-R~-~4-hy~roxybenzamido)perhydro~zepine trifluoro~cetic acid A solution of 0.070 g (0.071 mmole) of trans-N-benzyl-4-~(((3-benzyloxycarbonyl) -2-pyridinyl) carbonyl) -3, 5-dibenzyloxybenzoyloxy]-3-(4-benzyloxybenzamido)perhydroazepine in 8 ml of ethanol/methylene chloride (1:1) was treated with 10 L (0.142 mmole) of trifluoroacetic acid. The solution was stirred at room temperature for fifteen minutes. The solvent was evaporated and the ethanol/methylene chloride solvent was added twice more and evaporated in order to remove the excess trifluoroacetic acid. The residue was taken up in 10 ml of absolute ethanol and cooled to 0 C under nitrogen, and 0.030 g (0.025 mmole) of palladium hydroxide on carbon was added.
The reaction mixture was stirred under an atmosphere of hydrogen for twenty-four hours. The reaction mixture was filtered, evaporated and the residue was chromatographed on a 21 X 250 mm C18 column (solvent A: 95: 5 water/acetonitrile +
0.1% TFA; solvent B: 100% acetonitrile; gradient: 0 - 50% B
over 60 min., flow 15 ml/min). The pure fractions were pooled and evaporated to yield 0.010 g of a yellow powder, mp 198-205 C. Anal. Calcd for C27H25N30~-2H20-1.8TFA: C, 47.32; H, 4.00; N, 5.41. Found: C, 47.27; H, 3.86, N, 5.47.
W094/2~62 215 7 4 1 ~ PCT~S94/02283 ~+)-Trans-3-~-Hydroxy)benzamido-~ 3,5-~ihydroxy)-~-phenylcarbonyl]benzoyloxyperhydroazepine trifluoroacetic acid salt ~COMPOUND 544) OBn H~ O OOn O OH
Bn (~)-trans ~X~ ~F OH
1. (COC1)2 ,_~ N~f H2/Pd(OH)2 2. Et3N,DMAP ~ ~ (--~-` O
O OBn. Bn ~-~rans H CF3COOH
COMPOIJN`D 627 To a mixture of N-benzylated azepine intermediate (0.3 g, 0.697 mmol), Et3N (352.6 mg, 487 ~1, 3.485 mmol), and DMAP
(17.0 mg, 0.139 mmol) in CH2Cl2 (5 ml) was added a freshly made solution of benzophenone acid chloride (corresponding benzophenone acid: 336 mg, 0.766 mmol; oxalyl chloride: 2.0 M
solution in CH2C12, 0.697 ml, 1.394 mmol and cat. DMF) in CH2Cl2 (5 ml) at room temperature. The reaction mixture was stirred at room temperature overnight. Flash chromatography of the reaction mixture on silica gel using 3:2/Hexane:EtOAc as an eluent gave white solid product (469 mg, 71%).
CO~POUND 5~
The preceeding compound (300 mg, 0.315 mmol) in EtOAc:MeOH (1:1, 25 ml) was treated with CF3COOH (37.76 mg, W O 94t20062 2 ~ 5 7 ~ 12 PCTrJS94/02283 25.5 ~Ll, 0.33 mmol) and 20% Pd(OH)2 on activated carbon (150 mg, 50% on weight basis). The mixture was subjected to hydrogenolysis at 45 psi for 15 hr. The crude product after filtration and concentration was taken into DMF (0.5 ml) and purified on Cl8-HPLC column eluting with 596-50% acetonitrile in H2O containing 0.1% CF3COOH. The title compound was obtained as white powder (118 mg, 62%). m.p. 204-206 (dec)-C; 1HNMR
(DMSO-d6) ~ 8.42 (d, lH, NHCO), 7.58 (d, J = 7.5 Hz, 2H, ArH), 7.28-7.15 (m, 4H, ArH), 6.82 and 6.80 (s and s, 2H, ArH), 6.72 (d, J = 8.1 Hz, 2H, ArH), 5.18 (s, br, lH, CH--4), 4.45 (s, br, lH, CH-3), 3.30 and 3.10 (s and s, br, 4H, CH2N-2,7), 2.10-1.70 (m, 4H, CH2-5,6); IR (KBr) cm 1 3429, 1717, 1703, 1637, 1608, and 1509. Anal. Calcd. for C27H26N2O7 - 1.5 H2O - 1.0 TFA: C, 55.15; H, 4.79; N, 4.44. Found: C, 54.97; H, 5.08; N, 4.06.
_ WO94/2~K2 21~ 7 412 PCT~S94/02283 Trans-2- ~-Benzoyl-3, 5-dihy~roxybenzoyloxy) -1-hydroxybenzamido) cyclohexane ~COMPO~IND 545) Trans-2-(4-benzoyl-3,5-dibenzyloxybenzoyloxy)-1-(4-benzyloxybenzamido)-cyclohexane (172 mg, 231 ~mol) was dissolved in ethyl acetate (5 ml) and added to a stirring mixture of Pearlman's catalyst (Pd(OH) 2, 17 mg) in ethyl acetate (3 ml). The flask was evacuated under house vacuum and filled with H2(g) via balloon. After 3 h, additional Pearlman's catalyst (17 mg in 2 ml of EtOH) was added and left to stir overnight. TLC showed complete reaction (Starting material Rf=0.90, product Rf=0.41 in 50% EtOAc/hexanes.) The reaction mixture was filtered through Celite and concentrated to give the product as a yellow glass. The glass was triturated with water to give a pale yellow powder (122 mg, 86%): lH-NMR (DMSO, 300 MHz) ~ 1.27-1.57 (4H, m), 1.63-1.77 (2H, s), 1.84-1.89 (lH, m), 2.05-2.17 (lH, m), 4.05-4.18 (lH, m), 4.91-5.01 (lH, m), 6.75 (2H, d, J = 8 Hz), 6.97 (2H, s), 7.24-7.36 (2H, m), 7.48 (3H, t), 7.69 (2H, d, J = 8 Hz), 8.10 (lh, d, J = 9 Hz), 9.85 (lH, s), 9.95 (2H, s): 13C NMR (DMSO, 300 MHz) ~ 24.14, 24.52, 24.57, 31.20, 31.79, 51.73, 75.70, 107.68, 108.19, 114.98, 125.96, 126.29, 126.35, 128.35, 129.11, 129.15, 129.45, 132.08, 133.80, 137.24, 155.75, 160.29, 165.47, 166.13, 194.87. IR (KBr) cml 3360, 3271, 2943, 2858, 2360, 1708, 1659, 1610, 1541, 1507, 1450, 1424, 1369, 1347, 1278, 1240, 1177, 1106, 1047, 1011, 847, 771, 595. Anal. Calcd. for C27H2sNO7 - H20: C, 65.71; H, 5.51; N, 2.84. Found: C, 66.16: H, 5.49; N, 2.76.
W O 94/20062 215 7 ~12 PCTrUS94/02283 Anti-3-(~-benzyloxybenzami~o)-~-[3,5-dibenzyloxy-~-~3,~-dibenzyloxyphenylcarbonyl)benzoyloxy]-N-benzylperhy~roazepine ~CONPOUND 546) 3,5-Dibenzyloxy-4-[(3,4-dibenzyloxy)benzoyl]benzoic acid (500 mg, 0.768 mmol) was dissolved in anhydrous dichloromethane (6 ml). Anhydrous dimethylformamide (0.10 ml) was then added to the solution followed by oxalyl chloride (2 N in dichloromethane, 0.50 ml, 0.99 mmol). This solution was stirred for 1 h and then concentrated in vacuo. The resulting yellow oil was placed under high vacuum for a period of 1 h to make sure all of the excess oxalyl chloride was removed. The residue was dissolved in anhydrous dichloromethane (S ml), and a solution of dimethylaminopyridine (67 mg, 0.5S0 mmol), triethylamine, and anti-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine (SPC-103853, 215 mg, 0.500 mmol) in anhydrous dichloromethane (5 ml) was added under nitrogen. The reaction was stirred at room temperature for 1.5 h. At this point dichloromethane (100 ml) and sodium hydroxide (0.5 N in water, 30 ml) were added to the reaction. The aqueous and organic layers were separated and the organic phase was washed with brine (100 ml). The organic phase was isolated and dried with magnesium sulfate. The magnesium sulfate was filtered off, and the solution was concentrated in vacuo to yield a yellow solid.
The yellow solid was flash chromatographed on silica gel eluting with hexanes:ethyl acetate/9:1, 4:1, and 1:1 to yield a white solid of the title compound (400 mg, 75%): mp 65-C: lH
NMR (CDCl3) ~ 1.80 (m, 2H, CH2), 2.04 (m, 2H, CH2), 2.73 (m, 2H, NCH2), 3.01 (m, 2H, NCH2), 3.59 (d, J = 12.5 Hz, lH, NCH2Ph), 3.80 (d, J = 12.9 Hz, lH, NCH2Ph), 4.32 (m, lH, CH), 5.03 (s, 4H, CH2Ph), 5.12 (s, 2H, CH2Ph), 5.13 (s, 2H, CH2Ph), 5.22 (m, lH, CH), 5.24 (s, 2H, CH2Ph), 6.74 (d, J = 7.8 Hz, lH, NH), 6.86-7.56 (m, 39H, ArH); IR (KBr) cm 1580 (COO-), 1654 (CO).
Analysis calculated for C69H62N2O~: C, 77.95; H, 5.88; N, 2.63.
Found: C, 77.70; H, 5.99: N, 2.60.
W094/20062 21~ 7 412 PCT~S94/02283 Anti-~-[3~s-dihydroxy-~-~3~-dihydroxyphenylc~rbonyl)]
benzoyloxy-3-l~-hy~roxybenz~mido)perhydroazepine (COMPOUND 547) Anti-3-(4-benzyloxybenzamido)-4-[3,5-benzyloxy-4-(3,4-dibenzyloxyphenylcarbonyl)benzoyloxy]-N-benzylazepine (240mg, 0.23 mmol) and acetic acid were dissolved in methanol:ethyl acetate/2:1 in a 500 ml Parr bottle. Next, 5~ palladium on activated carbon (45 mg) was added under nitrogen. The reaction mixture was placed on a Parr hydrogenator for 3 h.
The mixture was then filtered over celite, and the filtrate was concentrated in vacuo to yield a yellow solid. The solid was flash chromatographed on silica gel eluting with dichloromethane:methanol/8:2 to yield a yellow solid of the title compound (57 mg, 48%): mp 176-C; H NMR (D6 DMSO) ~ 1.65 (m, lH, CH2), 1.78 (m, lH, CH2), 1.90 (2, 2H, CH2), 2.85 (m, 4H, N(CH2)2), 4.20 (m, lH, CH), 5.18 (m, lH, CH), 6.70-7.70 (m, 9H, ArH), 8.16 (d, J = 8.4 Hz, lH, NH); IR (KBr) cm1 1607 (CO), 1704 (COO-), 3431 (OH). Anal. calcd. for C27H26N2O8 H20: C, 59.50; H, 5.27; N, 5.14. Found: C, 59.54; H, 5.33; N, 4.93.
~1 L ~'7 W094/2~62 ~lJ J ~ PCT~S94102283 Trans-3-t3,5-Dihy~roxy-~-(2-hydroxy)phenylc~hQnyl]benzami~o-4-~-hydroxy)benzoyloxyperhy~ro~zepine ICOMr~u~v 548) To a solution of trans-N-benzyl-3-[3,5-dibenzyloxy-4-(2-benzyloxy)phenylcarbonyl]benzamido-4-(4-benzyloxy)-benzoyloxyazepine (65 mg, 0.068 mmol) in EtOAc/MeOH (5/15 ml) was introduced Pd(OH)z on carbon (35 mg of 20% Pearlman's catalyst). The reaction mixture was subjected to hydrogenolysis at 45 psi for 24 h at room temperature. The catalyst was filtered off through a pad of celite and washed with MeOH. The combined filtrate was concentrated to dryness and purified by flash column chromatography (SiO2: 60 ml, eluted with 10% to 20% MeOH in CH2C12). The title compound was obtained as a yellow solid (17 mg, 50%): mp 172-175-C, 1H NMR
(CD30D) ~: 7.65 (d, J = 8.7 Hz, 2H, ArH), 7.26 (td, lH, ArH), 7.10 (dd, lH, ArH), 6.74 (dd, lH, ArH), 6.60 (td, lH, ArH), 6.59 (d, J = 8.7 Hz, 2H, ArH), 6.50 (s, 2H, ArH), 5.03 (m, lH, CH-4), 1.19 (m, lH, CH-3), 2.95-2.70 (m, 4H, CH2N-2,7), 1.91-1.60 (m, 4H, CH2-5,6); IR (KBr) cm1 3434, 1700, 1623, 1610, 1542; low resolution FAB: (M + 1) 507.
_ W094/2~62 215 7 412 PCT~S94/02283 Tr~ns-3-t3,5-~ihy~roxy-4-~2-hy~roxyphenylra-honyl)]benz~mido-~-(4-hydroxy)benzoyloxy-N-isopropylperhydroazepine ~COMPOUND 549) To a solution of trans-N-benzyl-3-[3,5-dibenzyloxy-4-(2-benzyloxy)phenylcarbonyl]benzamido-4-(4-benzyloxy)-benzoyloxyperhydroazepine (190 mg, 0.199 mmol) in EtOAc-MeOH
(1:1, 30 ml) was introduced Pearlman's catalyst 20% Pd(OH)2 (70 mg). The mixture was subjected to hydrogenolysis at 50 psi for 17 hours. The catalyst was carefully filtered off through a pad of celite and washed with MeOH. The filtrate was concentrated and chromatographed on silica gel, eluting with 5%
MeOH in CH2C12. The title compound was obtained (500 mg, 50%);
the Parr bottle was contaminated with acetone and acid. All spectra ( H, C, APT, DEPT, MS) and CHN analysis fully support the structure. lH NMR (DMSO-d6) ~ 11.95 (s, brs, lH, OH), 10.25 (s, lH, OH), 10.00 (s, 2H, 20H), 8.20 (d, lH, NH), 7.78 (d, J
= 8.67 Hz, 2H, ArH), 753 (td, lH, ArH), 7.26 (dd, lH, ArH), 6.99 (d, lH, ArH), 6.87 (t, lH, ArH), 6.82 (d, J = 8.73 Hz, ArH), 6.74 (s, 2H, ArH), 5.01 (s, br, lH, CH-4), 4.18 (s, br, lH, CH-3), 2.90 (s, br, lH, CH(CH3)3), 2.68 (s, br, 4H, CH2N-2,7), 1.90 (s, br, 3H, CH2-6, CH-5), 1.63 (s, br, lH, CH-5), 1.00 (dd, 6H, 2CH3); 13C NMR (DMSO-d6 + D2O) ~ 55.02 (CH-(CH3)3), 18.66 and 18.40 (CH3)2CH); high resolution FAB M +
1: 549.2224; calculated for C30H32N2Oe: 549.2228. Anal. calcd.
for C30H32N2O~ 1.25 H2O: C, 63.09; H, 6.09; N, 4.90. Found: C, 63.06; H, 5.95; N, 4.66.
WOg4/2~62 21~ 7 4 ~ 2 PCT~S94/02283 --anti-4- ~3,5-Dihy~roxy-~- ~2-hy~roxybenzoyl)-benzoyl~mino)hexahy~ro-3-(4-hy~roxybenzoylamino)azepine, complex with water:acetonitrile ~1:1.7:0.3) (COMPOUND 550) ~8YNTg~8I8 OF COMPOUND 550) 8yn-3-Aminohexahy~ro-~-hydroxy-l-phenylmethylazepi~-2-one A solution of 3-acetylaminohexahydro-1-phenylmethyl-azepin-2,4-dione (0.82 g, 3.0 mmol) in absolute ethanol (15 ml) was treated with sodium borohydride (0.23 g, 6 mmol) and stirred for 30 min, then treated with water (5 ml) and concentrated in vacuo. The aqueous residue was extracted with methylene chloride (3 x 25 ml) and the combined organic extracts were dried (Na2SO~), concentrated in vacuo, and taken up in 2:1 ethanol/water (7.5 ml). Concentrated hydrochloric acid (2.5 ml) was added. The mixture was refluxed for 2 h and partially concentrated, then diluted with water (25 ml). The aqueous acidic mixture was extracted with ether (25 ml). The aqueous solution was basified with 30% sodium hydroxide and extracted with methylene chloride (3 x 40 ml). The combined methylene chloride extracts were washed with water (25 ml), dried (Na2SO~), and concentrated in vacuo to a yellow solid, which was recrystallized from ethyl acetate to afford syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.42 g, 60%) as a white solid.
8yn-3-Aminohexahydro-~-hydroxy-1-phenylmethylazepine A cooled (5-C) solution of lithium aluminum hydride/tetrahydrofuran (Aldrich, 1.0 N, 5.1 ml) under nitrogen was treated with syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.40 g, 1.7 mmol) in portions so that the pot temperature did not exceed 15-C. The mixture was refluxed for 6.5 h, cooled on an ice bath, and carefully treated with water (0.21 ml), 15~ sodium hydroxide (0.21 ml), and water (0.63 ml). The suspension was allowed to stir for 5 days (optimal time is 2-5 hours). The suspension was filtered, and the filtrate was concentrated in vacuo and chromatographed _ W094/2~62 215 7 412 PCT~S94/02283 on silica gel (eluted with 90:8:2 methylene chloride/methanol/
triethylamine). The appropriate fractions were concentrated in vacuo to afford syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.22 g, 58%) as a colorless oil.
8yn-~exahydro-~-hy~roxy-3-(4-phenylmethoxy)benzoyl~mino-1-phenylmethyl~zepine A solution of 4-benzyloxybenzoic acid (0.183 g, 0.8 mmol) in anhydrous tetrahydrofuran (2 ml) and N,N-dimethylformamide (0.5 ml) was treated with N,N'-carbonyldiimidazole (O.lS g, 0.9 mmol) and stirred at room temperature for 1.5h. The solution was treated with syn-3-aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.20 g, 0.9 mmol) in anhydrous tetrahydrofuran (1 ml). The mixture was stirred for 18 h, then concentrated in vacuo. The residue was taken up in lN sodium carbonate (20 ml), and the aqueous mixture was extracted with toluene (2x25 ml) cont~ining a little 2-propanol. The combined organic extracts were dried (Na2S0~) and the concentrated residue was flash chromatographed on silica gel (eluted with 3:1 ethyl acetate/hexane) to afford syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.17 g, 50%) as a viscous oil.
syn-~-Aminohex~hydro-3-(~-phenylmethoxy)benzoyl~mino-1-phenylmethylazepine A solution of syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)-benzoylamino-l-phenylmethylazepine(0.645g,1.5 mmol) in anhydrous tetrahydrofuran (3 ml) under nitrogen was cooled (ice bath, 5-C) and treated with triphenylphosphine (0.495 g, 1.8 mmol) and diethylazodicarboxylate (0.313 g, 1.8 mmol). The mixture was then treated dropwise over 15 min with a solution of diphenylphosphoryl azide (0.495 g, 1.8 mmol) in anhydrous tetrahydrofuran (3 ml). The reaction was stirred at room temperature for 18 h, then concentrated in vacuo, dissolved in a little methylene chloride, and passed through a short column of silica gel (eluted with 10% acetone/methylene chloride). The early fractions containing chromophoric W094/2~62 215 7 ~1~ PCT~S94/02283 material were concentrated in vacuo and dissolved in ethanol/acetic acid/water (6:1:1, 12 ml), then treated with zinc powder (0.50 g, 7.5 mmol). After 30 min, the mixture was filtered and the filtrate was concentrated in vacuo. The residue was taken up in 1.0 N sodium hydroxide (40 ml), and the aqueous mixture was extracted with methylene chloride (3 x 35 ml). The combined extracts were dried (Na2SO4) and concentrated in vacuo to afford crude syn-4-aminohexahydro-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepine (0.76 g).
syn-4-~3,5-Bis~phenylmethoxy)-~-~2-phenylmethoxybenzoyl))-benzoyl~minohexahydro-3-~-phenylmethoxy)benzoyl~mino-1-phenylmethylazepine ~COMPOUND 628) A solution of 2,2',6-tribenzyloxybenzophenone-4-carboxylic acid (0.545 g, 1.0 mmol) in anhydrous methylene chloride (3 ml) and N,N-dimethylformamide (0.1 ml) under nitrogen was treated with 2.0 N oxalyl chloride/methylene chloride (0.7 ml, 1.4 mmol) and stirred for one hour. The solution was concentrated in vacuo to a yellow solid, which was kept under high vaccuum for one hour. A solution of crude syn-4-aminohexahydro-3-(4-phenylmethoxy)-benzoylamino-1-phenylmethylazepine (0.76 g, from above) in methylene chloride (5 ml) was added to the acid chloride above, followed by 1.0 N
sodium hydroxide (4 ml). The mixture was stirred for 1.5 h, diluted with methylene chloride (15 ml) and separated. The aqueous layer was extracted with methylene chloride (15 ml) and the combined organic solution was washed with saturated aqueous sodium chloride, dried (Na2SO~), and concentrated in vacuo. The residue was chromatographed on silica gel (eluted initially with 2.5% acetone/methylene chloride, then with 7.5%
acetone/methylene chloride) to afford syn-4-(3,5-bis(phenylmethoxy)-4-(2-phenylmethoxy-benzoyl))-benzoylaminohexahydro-3-(4-phenylmethoxy)-benzoylamino-1-phenylmethylazepine (0.52 g, 54%) as an opaque gum.
_ WO 94/20062 21 S 412 PCT/US94/02283 syn~ 3, 5-Dihydroxy-~- ~ 2-hydroxybenzoyl ) -benzoylamino)hexahy~ro-3-14-hydroxybenzoylamino)azepine, complox with water:acetonitril~ ~1:1.7:0.3) (COMPO~ID 550) A solution of syn-4- (3,5-bis (phenylmethoxy) -4- (2-phenylmethoxybenzoyl ) ) benzoylaminohexahydro-3 - (4 -phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.39 g, 0.41 mmol) in ethanol/ethyl acetate (1: 1, 30 ml) was placed in a Parr bottle and treated (under nitrogen) with Pearlman ' s catalyst (Aldrich, 150 mg), then subjected to hydrogenation in a Parr apparatus for 18 h at 48-50 psi. The reaction mixture was carefully purged of hydrogen and the solution was filtered through celite (care taken not to let filter cake dry). The filtrate was concentrated in vacuo to afford crude material, which was chromatographed on silica gel (eluted with 2 :1 methylene chloride/isopropanol) to give syn-4- (3,5-dihydroxy-4-( 2 -hydroxybenzoyl ) benzoylamino ) hexahydro-3 - ( 4 -hydroxybenzoylamino)azepine, complex with water:acetonitrile (1:1.7:0.3) (0.055 g, 27%) as a yellow solid, which was triturated with acetonitrile to afford a yellow powder: mp 210 - 215-C; Rf (2: 1 methylene chloride/isopropanol on silica) 0.50; IR (KBr) 1624 cm 1; lH NMR (d6-DMSO) ~ 8.33 (d, lH, J = 7 Hz), 7.97 (d, lH, J = 7 Hz), 7.73 (d, 2H, J = 9 Hz), 7.55 (m, lH), 7.30 (m, lH), 7.00 (m, lH), 6.89 (m, lH), 6.84 (s, 2H), 6.82 (d, 2H, J = 9 Hz), 4.25 - 4.40 (m, 2H), 3.00 - 3.10 (m, 2H), 2.80 - 3.00 (m, 2H), 1.85 - 1.95 (m, lH), 1.65 - 1.80 (m, 3H). Anal. calcd. for C2~H27N307 1.7H20 0.3(CH3CN): C, 60.44;
H, 5.75; N, 8.43. Found: C, 60.34; H, 5.56; N, 8.42.
W094/20062 215 7 41~ PCT~S94/02283 Hexahydro-3-(~-phenylmethoxy)benzoylamino-1-phenylmethyl-azepin-~-one (COMPOUND 551) A 25 ml 3-neck round bottom flask under nitrogen was charged with 2.0 N oxalyl chloride/methylene chloride (Aldrich, 1.125 ml, 2.25 mmol), diluted with anhydrous methylene chloride (2 ml), cooled (-65-C), and treated dropwise with anhydrous dimethylsulfoxide (0.35 g, 4.5 mmol) in anhydrous methylene chloride (1.2 ml) at a rate to keep the pot temperature below -60-C. The mixture was stirred at -65 + 5C for 30 min, then treated dropwise with a solution of syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.645 g, 1.5 mmol) in anhydrous methylene chloride (1.5 ml) at a rate to keep the pot temperature below -55-C. The mixture was stirred at 55 + 5-C for 2 h, then treated dropwise with triethylamine (1.5 ml), warmed to room temperature over 1 h, and diluted with methylene chloride (10 ml). The organic solution was washed with water (10 ml), saturated aqueous sodium bicarbonate (10 ml), dried (Na2S0~), and concentrated in vacuo. The residue was chromatographed on silica gel (eluted with 5% acetone/methylene chloride) to afford hexahydro-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepin-4-one (0.53 g, 82%) as a viscous colorless oil.
~-Aminohexahydro-3-(~-phenylmethoxy)benzoylamino-1-phenyl-methylazepine, 1:1 mixture of syn and anti i~omers A solution of hexahydro-3-(4-phenylmethoxy)-benzoylamino-l-phenylmethylazepin-4-one (0.37 g, 0.86 mmol) in ethanol (5 ml) was treated with hydroxylamine hydrochloride (80 mg, 1.15 mmol) and 25% methanolic sodium methoxide (Aldrich, 70 mg, 0.32 mmol) and heated to 50-C for 1 h. The mixture was cooled to room temperature, treated with additional 25% sodium methoxide (0.18 g, 0.83 mmol), then stirred for 10 min and partially concentrated in vacuo. The residue was taken up in 0.5 N sodium hydroxide (5 ml) and extracted with methylene chloride (3 x 15 ml). The combined organic extracts were washed with saturated aqueous sodium chloride, dried (Na2SO4), and concentrated in vacuo. This residue was dissolved in 95%
W094/20062 2 1 ~ 7 4 ~ ~ PCT/US94/022~
ethanol (20 ml), treated with Raney Nickel (Aldrich, 1/4 teaspoon) in a Parr bottle, and subjected to hydrogenation at 30 psi for 2.25 h (more time needed). The solution was carefully evacuated of hydrogen, filtered through celite (do not allow to dry), and the filtrate was concentrated in vacuo and chromatographed on silica gel (eluted first with 15%
methanol/methylene chloride, then with 25%, and finally with 33%) to afford 4-aminohexahydro-3-(4-phenylmethoxy)benzoyl-amino-l-phenylmethylazepine, 1:1 mixture of syn and anti isomers (0.15 g, 40%) as a viscous colorless oil which was stored under nitrogen.
~nt~-4-~3,5-B$~phenylmethoxy)~ 2-phenylmethoxybenzoyl))-benzoyl~minohexahydro-3-(~-phenylmQthoxy)benzoyl~mino-l-phenylmethylazepine ~COMPO~ND 629) A solution of 2,6,2'-tribenzyloxybenzophenone-4-carboxylic acid (0.245 g, 0.45 mmol) in anhydrous methylene chloride (1.5 ml) under nitrogen was treated with N,N-dimethylformamide (3 drops), then with 2.0 N oxalyl chloride/methylene chloride (0.30 ml, 0.60 mmol). The vigorous bubbling soon subsided and the mixture was stirred for 1 h, concentrated in vacuo, and kept under high vacuum for 1 h to ensure removal of excess oxalyl chloride. The acid chloride was dissolved in methylene chloride (2.5 ml) and treated with 4-aminohexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine, 1:1 mixture of syn and anti isomers (0.14 g, 0.325 mmol) followed by 1.0 N sodium hydroxide (1.5 ml).
The biphasic mixture was rapidly stirred for 2 h and separated.
The aqueous layer was extracted with methylene chloride (2 x 7 ml), and the combined organic extracts and organic layer were washed with saturated aqueous sodium chloride (10 ml), dried (Na2SO~), and concentrated in vacuo. Flash chromatography on silica gel (eluted first with 3% acetone/methylene chloride, then with 7%, and finally with 10%) afforded anti-4-(3,5-bis(phenylmethoxy)-4-(2-phenylmethoxy-benzoyl))-benzoylaminohexahydro-3-(4-phenylmethoxy)benzoylamino-1-WO 94/20062 215 7 il ~ 2 PCT/US94/02283 phenylmethylazepine (0.13 g, 42%) as a colorless foam, followedby the syn isomer (0.13 g, 42%) .
~nti-4-~3,5-Dihydroxy-~-~2-hydroxybenzoyl)benzoyl~mino)hexa-hydro-3-~-hydroxybenzoylamino)azepine, complex with water:2-prop~nol ~1:1.5:0.~) ~COMPOUND 551) A solution of syn-4- (3,5-bis (phenylmethoxy) -4- (2-phenylmethoxybenzoyl) ) benzoylaminohexahydro-3- (4-phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.13 g, 0.136 mmol) in ethanol/ethyl acetate (1: 1, 25 ml) was placed in a Parr bottle and treated (under nitrogen) with Pearlman ' s catalyst (Aldrich, 120 mg), then subjected to hydrogenation in a Parr apparatus for 18 h at 50 psi. The reaction mixture was carefully purged of hydrogen and the solution was filtered through celite (care taken not to let filter cake dry). The filtrate was concentrated in vacuo to afford crude material, which was chromatographed on silica gel (eluted with 1: 1 methanol/methylene chloride). This concentrated product was dissolved in 2-propanol, filtered to remove inorganic material, re-concentrated in vacuo, and triturated from ether to afford anti-4- ( 3, 5-dihy~roxy-4- ( 2 -hydroxybenzoyl ) -benzoylamino) hexahydro-3- (4-hydroxybenzoylamino) azepine complex with water:2-propanol (1:1.5:0.4) (56 mg, 75%) as a yellow powder: mp 207 - 210 C; Rf (2: 1 methylene chloride/2-propanol on silica) 0.20: IR (KBr) cm 1 1639, 1607; H NMR (d6-DMSO) 8.32 (d, lH, J = 7 Hz), 7.96 (d, lH, J = 7 Hz), 7.66 (d, 2H, J
= 9 Hz), 7.52 (m, lH), 7.27 (m, lH), 6.97 (m, lH), 6.86 (m, lH), 6.77 (d, 2H, J = 9 Hz), 6.72 (s, 2H), 4.00 - 4.15 (m, 2H), 2.60 - 3.00 (m, 4H), 1.50 - 1.90 (m, 4H) . Anal . calcd. for C2~Hz~N307 1-5 H20 0.4 (C3H80): C, 60.85; H, 6.01; N, 7.55.
Found: C, 60.68; H, 5.68; N, 7.22.
~15~ 2 - PCT~S94/0~R~
~ Anti-3-~ minobenzam~do)-~-[3,5-dihy~roYy-~-(2-hy~roxyphenylcarbonyl)]benzyloxyperhy~roazepine (COMPOUND552) OE~ 8110 OH HO
OH H ` O~ ~ o (';;)-- ,C, ~ N2 ~ C~ NOz -- (;~-- ~ C~ NH2 3,5-dibenzyloxy-4-~(2-benzyloxybenzoyl]benzoic acid (218 mg, 0.40 mmol) was dissolved in anhydrous dichloromethane (2.0 ml), and dimethylformamide (0.05 ml). The reaction was placed under nitrogen, and oxalyl chloride (0.3 ml, 2M in dichloromethane) was slowly added via a syringe. The reaction was stirred at room temperature for 1 h and then concentrated in vacuo and placed under high vacuum for 1 h. The residue was redissolved in anhydrous dichloromethane (2 ml) and stirred under nitrogen. Next, a solution of (+)-anti-3-(4-nitrobenzamido)-4-hydroxybenzylperhydroazepine (120 mg, 0.32 mmol) and dimethylaminopyridine (44 mg,0.36 mmol) in triethylamine (0.9 ml) and anhydrous dichloromethane (3 ml) were added. The reaction was then stirred for 1 h at room temperature. At this point, dichloromethane (50 ml) and 0.5 N
sodium hydroxide (20 ml) were added, and the reaction was transferred to a separatory funnel. The aqueous layer was removed, and brine was added. The organic layer was then isolated, dried over sodium sulfate, filtered, and concentrated in vacuo to yield a yellow oil. The oil was flash chromatographed eluting with hexane:ethyl acetate/4:1. Pure fractions of the major product were concentrated in vacuo to yield 180 mg of white solid (COMPOUND 636). The solid was then dissolved in methanol:ethyl acetate/2:1 (30 ml) and acetic acid ~094/20062 2 1 ~ 7 4 1 2 PCT~S94/02283 (0.2 ml) in a 500 ml Parr bottle along with 5% palladium on activated carbon (70 mg). The Parr bottle was then placed on the Parr hydrogenator for 24 h. The reaction mixture was filtered to remove the catalyst, and then concentrated in vacuo to yield a yellow solid. The solid was flash chromatographed eluting with dichoromethane:methanol/9:1 to yield a yellow solid of the title compound (60 mg, 61%): mp 175 - 185-C; NMR
(CD30D) consistant with structure; IR (RBr) cm1 3389 (OH); 2361 (alkyl); 1704 (ester); 1625 (ketone). Anal. calcd. for C27H27N3O7 1.0 H2O 1.0 acetic acid: C, 59.69; H, 5.70; N, 7.20. Found: C, 59.94; H, 5.70; N, 7.31.
~09~/20062 21~7412 _ PCT~S94/0 ~ + ) -Anti-3- ( ~- f luoro~^n~mi~o)-4-t3,5-dihydroxy-~-(2-hy~roxy-phenylc~rbonyl)]benzyloxyperhydroazepine (COMPOUND 53) OBn i3nO OH HO
0,.~~ o ,~_,0,~
o~ F _ (;;)-- C~ F (;;~_ ~H OH F
3,5-Dibenzyloxy-4-t(2-benzyloxybenzoyl]benzoic acid (545 mg, 1.0 mmol) was dissolved in anhydrous dichloromethane (5.0 ml), and dimethylformamide (0.05 ml). The reaction was placed under nitrogen, and oxalyl chloride (1.0 ml, 2 M in dichloromethane) was slowly added via a syringe. The reaction was stirred at room temperature for 2 h and then concentrated in vacuo and placed under high vacuum for 1.5 h. The residue was redissolved in anhydrous dichloromethane (3 ml) and stirred under nitrogen. Next, a solution of (+)-anti-3-(4-fluorobenzamido)-4-hydroxy-N-benzylperhydroazepine (300 mg, 0.93 mmol) and dimethylaminopyridine (125 mg, 1.02 mmol) in triethylamine (0.9 ml) and anhydrous dichloromethane (2 ml) were added. The reaction was then stirred for 20 min at room temperature. At this point, dichloromethane (50 ml) and 0.5 N
sodium hydroxide (30 ml) were added, and the reaction was transfered to a separatory funnel. The aqueous layer was removed, and brine was added. The organic layer was then isolated, dried over sodium sulfate, filtered, and concentrated in vacuo to yield a light brown foam. The foam was flash chromatographed eluting with hexane:ethyl acetate/2:1. Pure fractions of the major product were concentrated in vacuo to yield 440 mg of white solid (COMPOUND 631). 260 mg of the W094/20062 2 1 5 7 4 ~ ~
PCT~S94/02283 solid was dissolved in methanol:ethyl acetate/5:1 (30 ml) and acetic acid (0.2 ml) in a 500 ml Parr bottle along with 5%
palladium on activated carbon (100 mg). The Parr bottle was then placed on the Parr hydrogenator for 4.5 h. The reaction mixture was filtered to remove the catalyst, and then concentrated in vacuo to yield a yellow solid. The solid was flash chromatographed eluting with dichoromethane:methanol/8:2 to yield a yellow solid of the title compound (110 mg, 72%):
mp 189-C; IR (KBr) cm1 3349 (OH); 2362 (alkyl); 1704 (ester);
1626 (ketone). Anal. calcd. for C27H25N2O7F 1.0 H2O 1.0 acetic acid: C, 59.38; H, 5.33; N, 4.78. Found: C, 59.09; H, 5.00; N, 4.71.
W094/20062 2 1 5 7 ~ 1 2 - PCT~S94/022 Tr~ns-3-~4-hy~,ox~z~m~do)-~-t~-(2-hy~oh~ 6-hy~ c~rhonyl-benzoyl)-3,5-~ihy~roxybenzoyloxyl]perhy~rothiepine (COMPOUND
55~) N~NHa 13~OH m~ DMF O~OTBDMS )O~H~ ~OH
HO~OH Ms Cl. EI~N ~AsO~OM~ L~MS EI~N TBDIA5~N, H NH~ 1) ~~ CO~ OBn ~
I.JAIH~THF ~ CDI, THF H ~ COCI
H~ ~N~
\_~S 2)lNN OH H~O ~S El~N DM~P,CH~
OBn OE~n H~ Pd(OHb~ ~(OH OH
l~nO~_ H ~ ~ OH THF HO ~_ ~N~ O ~ O
S
A mixture of 1,3-cyclohexadienemonoepoxide (9.61 g, 100 mmol, for preparation see: J. K. Crandall et àl., J. Org.
Chem., 1968, 33, 423), NaN3 (26 g, 400 mmol), NH4Cl (10.7 g, 200 W094/20062 215 ~ PCT~S94/02283 mmol), MeOH (300 ml), and H2O (50 ml) was stirred at 65 C for 16h. The resultant slurry was evaporated to remove all the volatile materials and the residue was treated with 1 N NaOH
(100 ml), and extracted with CH2C12 (3 x 60 ml). The combined CH2C12 extracts were washed with H20 (2 x 100 ml), dried (MgSO4), evaporated, and chromatographed on a SiO2 column (Et2O:hexane = 1:4) to give a colorless oil (7.308 g, 53%).
A mixture of the product of the previous reaction (7.23 g, 52 mmol), TBDMS-Cl (8.227 g, 55 mmol), and imidazole (5.308 g, 78 mmol) in N,N-dimethylformamide (50 ml) was stirred at room temperature for 16h. Et2O (250 ml) was added and the resultant mixture was washed with H2O (5 x 100 ml), dried (MgSO~), and evaporated to give a colorless oil (12.76 g, 97%) which was used in the next step without further purification.
A solution of oil (7.63 g, 30 mmol) in MeOH (150 ml) and CH2C12 (30 ml) was cooled to -78-C, stirred, and treated with a mixture of air and 03 until a blue color persisted (ca.
1.5h). Excess O3 was removed by bubbling N2 through the solution, and then NaBH~ (4.54 g, 120 mmol) was added. After being stirred at -78-C for 30 min. followed by 30 min. at room temperature the resultant mixture was poured into 1 N HCl (100 ml) and then evaporated to remove the volatile components. The aqueous residue was extracted with Et2O (4 x 50 ml), and the combined ether extracts were washed with HzO (100 ml), brine (100 ml), dried (MgSO~), evaporated, and crystallized from Et2O:hexane to give a white solid (6.772 g, 78%).
Methanesulfonyl chloride (2.67 ml, 34 mmol) was added dropwise to a stirred solution of the borohydride reduction product (4.01 g, 14 mmol) and Et3N (S.76 ml, 41 mmol) in CH2Clz (40 ml) at 5-C. After 30 min. the cooling bath was removed and stirring was continued for 16h. The resultant mixture was diluted with CH2C12 (20 ml), washed with 1 N HCl (3 x 30 ml), H2O (2 x 30 ml), brine (2 x 30 ml), dried (MgSO4), and evaporated. The residue was chromatographed (SiO2, Et2O:hexane = 1:1 followed by Et2O:hexane:CH2Cl2 = 1:1:0.25) to give a pale yellow oil (5.885 g, 96%).
UO9~/20062 2 1 5 7 ~ 1 2 PCT~S94/02283 A mixture of the product of the previous reaction (2.24 g, 5 mmol), lithium sulfide (253 mg, 5.5 mmol), and Et3N
(2.09 ml, 15 mmol) in EtOH (100 ml) was stirred under reflux for 16h. The resultant mixture was evaporated and the residue was chromatographed (SiO2, Et2O:hexane = 1:30) to give a colorless oil (515 mg, 36%).
A solution of LiAlH4 in Et2O (1 M, 3.58 ml) was added to the colorless oil from the reaction above (515 mg, 1.79 mmol) in Et2O (9 ml) and the mixture was stirred at room temperature for 2h. 5N aq NaOH (5 ml) was added cautiously and stirring was continued at room temperature for 16h. The phases were separated and the aqueous phase was saturated with NaCl and extracted with CH2C12 (3 x 10 ml). The combined organic layers were dried (MgSO4), and evaporated to give a colorless oil which solidified on standing (250 mg, 95%). This material was used in the next step without further purification.
A mixture of 4-benzyloxybenzoic acid (427 mg, 1.87 mmol) and l,l'-carbonyldiimidazole (302 mg, 1.87 mmol) in THF
(5 ml) was stirred at room temperature for 2h and then the product of the previous reaction (250 mg, 1.70 mmol) in THF (5 ml) was added. The mixture was stirred at room temperature for 36h, evaporated, dissolved in CH2C12 (30 ml), washed with H2O, and dried (MgSO~). The resultant solution was diluted with hexane (10 ml) and rotaevaporated at 0-C to ca. 10 ml. The white precipitate was collected, dried under vacuum, and dissolved in a mixture of MeOH (10 ml) and THF (8 ml). 1 N aq.
NaOH (1 ml) was added and the mixture was stirred at room temperature for 4h. The volatile components were evaporated and the residue was dissolved in CH2C12 (25 ml), washed with H2O
(3 x 5 ml), dried (MgSO4), and filtered. The filtrate was diluted with hexane (lO ml) and rotaevaporated at 0 C to ca. 5 ml. The precipitate was collected, washed with hexane, and dried under vacuum to give a white powder (382 mg, 67%).
To a solution of 4-(2-benzyloxy-6-benzyloxycarbonyl-benzoyl)-3,5-dibenzyloxybenzoic acid (305 mg, 0.45 mmol) and a drop of N,N-dimethylformamide in CH2C12 (1.3 ml) at 5C was w094~20062215 7 ~12 PCT/US94/02283 added a solution of oxalyl chloride in CH2Cl2 (2 M, 0.25 ml).
The resultant mixture was stirred at room temperature for 2h and then evaporated. The residue was dried under vacuum for 2h, dissolved in CH2C12 (0.8 ml), and added to a mixture of the above intermediate (143 mg, 0.4 mmol), Et3N (61 mg, 0.6 mmol), and 4-N,N- dimethylaminopyridine (5 mg, 0.04 mmol) in CH2Cl2 (1.2 ml). The resultant solution was stirred at room temperature for 17h, diluted with CH2C12 (10 ml), washed with H2O (3 x 5 ml), dried (MgSO~), and evaporated. The residue was chromatographed (SiO2, Et2O:hexane:CH2Cl2 = 1:1:0.5) to give a colorless oil (347 mg, 76%).
This intermediate (105 mg, 0.1 mmol), Pd(OH)2 on carbon (20 wt%, contains > 50% moist, 281 mg, 0.2 mmol), THF (1 ml), and MeOH (1 ml) was stirred and treated with 1 atm H2 at room temperature for 40h. The resultant mixture was filtered through a Florisil pad and the Florisil pad was washed with MeOH (15 ml). The combined filtrate and wash were evaporated to give a yellow solid (25 mg, 35%) (COMPOUND 554). IR (KBr, cm ): 1706, 1689, 1633.
1, 1-Dioxo-trans-3- ~ ~-hydroxybenz~mi~o ) -~- t ~.- ( 2-hy~roxy-6-hydroxycarbonylbenzoyl ) -3, 5-~lihy~roxybenzoyloxyl ] -perhydrothiepine ~COMPO~JND 555) ~o ~o CH,CO,H ~_ ~ ~CH,CO,H. ct'.,a, 11l1 ~ OH OH
H,. Pd~OH), C
C~so o Figure AA
Peroxyacetic acid (32 wt% in acetic acid, 37 mg, 0.155 mmol) was added to a solution of Compound 632 (75 mg, 0. 074 mmol, see Compound 554 for preparation) in CH2Cl2 (0.7 ml). The resultant mixture was stirred at room temperature for lh, diluted with CH2C12 (10 ml), and washed with sat. aq. K2CO3 ( 3 x 5 ml). The organic layer was dried (MgSO4) and evaporated to give a white solid which was recrystallized from hot EtOAc (10 ml, contains ca. 2 ml of THF and 3 ml of hexane) to give a white powder (61 mg, 79~) (COMPOUND 633).
21~7412 ~094/20062 PCT~S94/02283 A mixture of Compound 633 (61 mg, 0.058 mmol), Pd(OH)2 on carbon (20 wt%, contains > 50% moist, 8 mg, 0.0058 mmol), THF (1 ml), and MeOH (1 ml) was stirred and treated with 1 atm H2 at room temperature for 3 hr. The resultant mixture was filtered through Florisil and the filtrate was evaporated to give the title compound as a yellow solid (33 mg, 95~). IR
(KBr, cm ): 1718, 1686, 1635.
_ WO9~/20062 21~7~
Tr~ns-4-(4-(2-c~rboxy-6-hydroxybenzoyl)benzoyloxy)-3-(4-hy~roxybenz~mi~o)perhydro~zepine Trifluoro~cetic Aci~ ~lt Hy~r~te ~COMPOUND 556) OBn ( CO2H
~ t~ BuU ~ PDC, DMF
MeO~OMeO~SJ oan MeO~I OBn CO2Bn BnB- ~ KOH, DMF
KDCMO~3 MeO ~ OBn o CO2Bn BnO~
)~\ 1) (coa)2 ~, OBn "~/ 2) IPr2NEI, DMAP, ~/
HO2C OBn OH O ~~
(~ . N ~ OBn ~ H~ OBn Bn Bn 6~
HoO
H2 ~ OH
Pd(OHh . N~ OH
H
W094/20062 215 7 ~12 PCT~S94/02283 Methyl-4-(6-Benzyloxy-2-hy~roxymethylbenzoyl)benzoate To a solution of 1.07 g (5.00 mmol) of 3-benzyloxybenzyl alcohol in 15 ml of toluene at -5CC under an atmosphere of nitrogen was added 5.8 ml (12.2 mmol) of a 2.1 M solution of butyllithium in hexanes over 15 min. The solution was stirred at -5C for 6 h, after which it was cooled to -78-C, and a solution of 1.00 g (5.03 mmol) of 4-(methoxycarbonyl)benzoyl chloride in 5 ml of tetrahydrofuran was added. The mixture was stirred for 1 h, after which it was poured onto 200 ml of ether and 100 ml of saturated aqueous ammonium chloride, and this mixture was stirred for 10 min.
The layers were separated, and the organic phase was washed with saturated aqueous sodium bicarbonate and brine, dried over magnesium sulfate, and evaporated to give the crude product.
Flash chromatography on silica gel eluting with 3/1 ethyl acetate -hexane afforded 0.68 g (36~) of the title compound as a white solid, which was carried on to the next step.
Methyl-~-~6-Benzyloxy-2-ca,~6~benzoyl)benzoate To a solution of 0.63 g (1.7 mmol) of methyl 4-(6-benzyloxy-2-hydroxymethylbenzoyl)benzoate in 20 ml of dimethylformamide was added 4.41 g (11.7 mmol) of pyridinium dichromate. The solution was stirred at room temperature under a nitrogen atmosphere for 4 days, after which it was poured onto 300 ml of ether and washed with 200 ml of water, 150 ml of 2 M HCl, and 150 ml of brine, and dried over magnesium sulfate.
Evaporation of the solvent afforded 0.47 g (72%) of the crude product. This material was sufficiently pure for further use and was carried directly to the next step.
~ethyl-4-~6-Benzyloxy-2-~benzyloxyc~rbonyl)benzoyl)benzoate To a c ~ution of 0.47 g (1.2 mmol) of methyl 4-(6-benzyloxy-2-carboxybenzoyl)benzoate in 20 ml of dry dimethylformamide was added 501 mg (3.62 mmol) of potassium carbonate and 0.158 ml (227 mg, 1.32 mmol) of benzyl bromide.
The solution was stirred at room temperature under a nitrogen atmosphere for 18 h. The mixture was then poured onto 300 ml WO9-1/20062 215 7 ~12 PCT~S94/02~83 of ether and washed with two 200 ml portions of water and then with 150 ml of brine, and dried over magnesium sulfate.
Evaporation of the solvent afforded 0.57 g of the crude product, which was chromatographed on silica gel, eluting with 3/1 hexane-ethyl acetate to give 0.32 g (54%) of the title compound as a colorless oil. This material was used directly in the next step.
~-(6-Benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoic Acid A solution of 0.301 g (0.614 mmol) of methyl 4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoate in 7 ml of DMF
was treated with 0.337 ml of a 2 M aqueous solution of potassium hydroxide under an atmosphere of nitrogen for 20 h.
The mixture was then poured onto 100 ml of ethyl acetate and washed with 60 ml each of 0.2N HCl, water, and brine. The organic extracts were dried over magnesium sulfate and evaporated to give 0.32 g of the crude product, which was chromatographed on a 41 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA solvent B: 100% acetonitrile:
gradient: 0-100% B over 60 min, flow: 25 ml/min). The pure fractions were pooled and evaporated and then lyophilized from water to give 70 mg of the title compound as a white solid, which was carried on as is to the next step.
Tr~ns-N-Benzyl~ -(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl) benzoyloxy)-3-(~-benzyloxybenz~mido)perhydroazepine A solution of 68 mg (0.143 mmol) of 4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoic acid in 5 ml of methylene chloride containing a trace (approximately 1 ~L) of dimethylformamide was cooled to O-C. A 2.0 M solution of oxalyl chloride (0.11 ml, 0.22 mmol) was added, and the mixture was stirred under a nitrogen atmosphere for 2 h. An additional 0.17 ml of oxalyl chloride was added, and the mixture was stirred for an additional 2 h. The reaction mixture was evaporated, and the residue was evaporated twice from 10 ml of methylene chloride. The residue was dissolved in 3 ml of methylene chloride, and was added to a solution of 69.6 mg 21~1 2 (0.162 mmol) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine, 29.8 ,uL (0.17 mmol) of diisopropylethylamine, and 5.4 mg of DMAP in 5 ml of methylene chloride at O-C. The mixture was stirred at room temperature under a nitrogen atmosphere for 17 h, after which it was diluted with 30 ml of methylene chloride, washed with saturated sodium bicarbonate and brine, dried over magnesium sulfate, and evaporated to give 146 mg of the crude product. Chromatography on silica gel eluting with 1/1 hexane-ethyl acetate gave 46 mg (37%) of the title compound as a yellow oil, which was taken directly to the next step.
Trans~ (2-Carboxy-6-hy~roxybenzoyl)benzoyloxy)-3~
hydro~cybenzami~o)perhydroazepine Trifluoroacetic Acid Salt Hydrate A solution of 46 mg (0.052 mmol) of trans-N-benzyl-4-(4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoyloxy)-3-(4-benzyloxybenzamido)azepine in 10 ml of ethanol was treated with 8.1 ~L of trifluoroacetic acid, cooled to O-C, and 22 mg of moist 10% palladium hydroxide on carbon was added. The mixture was then stirred under an atmosphere of hydrogen for 19 h at room temperature. The mixture was filtered, evaporated, and the residue was chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min).
The pure fractions were pooled and evaporated and then lyophilized from water to give 9.8 mg (28%) of the title compound as a white fluffy solid. FABMS: m/z 519 (M + H).
Anal. Calc. for C28H26N208 2.5 H20 TFA; C, 53.18 H, 4.76; N, 4.13. Found: C, 53.43: H, 4.64; N, 4.30 Wo 94/20062 215 7 412 PCTtUS94tO2283 Trans 4 - t 4 - ~ 2 -Hy~roxycarbonyl - 6-hy~roxybenzoyl ) - 3, 5-dih~ oa~enzoyloxy~-3- ~4-hydlohyLe"z~ o) -1- ~phenyl~ulfonyl) -pyrrolidine ~COMPO~ND S62 ) CHzCIz H20 ~, OBn H-~FA S 02Ph C02Bn Pr~El, I OBn DMAP,~.
OBn Hæ~O~N~ EIOH. ~F ~ h OBn b~gH S02Ph BnO2Cb_OBn N
Figure AC
+ ) -Trans-3- ~ ~-benzyloxybenzamido) -~-hydroxy-l-~phenylsul f onyl ) pyrrolidine To a slurry of (+)-Trans-3-(4-benzyloxybenzamido)-4-hydroxypyrrolidine TFA (150 mg, 0.352 mmol) in H20 (8.8 ml) and CH2Cl2 (8.8 ml) were added anhydrous Na2C03 (112 mg, 3.0 eq, 1.06 mmol) then benzene sulfonyl chloride (58 ~1, 0.458 mmol, 1.3 eq), and the mixture stirred at room temperature 15 h. The solution was then diluted with CH2Cl2 (20 ml) and poured into H20 (20 ml) and methanol (4 ml). The layers were separated and the aqueous layer extracted with CH2C12 (3 x 30 ml). The organics were combined, dried (MgS0~), filtered and evaporated to a white powder (159 mg, quant yield): 1H NMR (CD30D) ~ 7.62 - (d, J = 7.7 Hz, 2H), 7.43 (d, J = 8.9 Hz, 2H), 7.35-7.30 (m, 2~57412 W094/20062 PCT~S94102283 3H), 7.25-7.10 (m, 5H), 6.80 (d, J = 8.8 Hz, 2H), 4.95 (s, 2H), 4.06-4.00 (m, lH), 3.95-3.90 (m, lH), 3.50-3.35 (m, 2H), 3.15 (dd, J = 10.6, 3.9 Hz, lH), 2.99 (dd, J = 10.8, 3.2 Hz, lH).
Trans-4-[4-~6-benzyloxy-2-~benzyloAyc~rbonyl)benzoyl]-3,5-dibenzyloAybenzoyloAy)-3- ~-benzyloxybenzamid,o)-l-(phenylsulfonyl)pyrrolidine (CONPO~nlD 639) To a solution of the previous product (159 mg, 0.352 mmol) in CH2Cl2 (6.0 ml) were added 4-dimethylaminopyridine (43 mg, 0.352 mmol, l.o eq), diisopropylethylamine (74 ~1, 0.42 mmol, 1.2 eq) then a solution of acid chloride (0.383 mmol, 1.1 eq) in CH2Cl2 (3.0 ml). The mixture was stirred at room temperature under N2 14 h. The reaction mixture was then diluted with CH2C12 (30 ml), and washed with 10% NaHCO3 (50 ml) then brine (50 ml). The aqueous layers were combined and extracted with CH2C12 (2 x 50 ml). The organics were combined, dried (MgSO~), filtered and evaporated. Flash column chromatoghraphy of the residue (2:1 hexane:ethyl acetate) on silica gel provided the title compound (183 mg, 47%): lH NMR
(CDCl3) ~ 7.77 (d, J = 6.7 Hz, 2H), 7.70 (d, J = 8.8 Hz, 2H), 7.47-7.16 (m, 14H), 7.15-7.05 (m, 6H), 7.04-6.94 (m, 3H), 6.90-6.83 (m, 3H), 6.42 (d, J = 6.7 Hz, lH), 5.30 (dt, J = 5.1, 2.6 Hz, lH), 5.18 (s, 2H), 5.13 (s, 2H), 4.78 (m, 2H), 4.76 (s, 2H), 4.72 (s, 2H), 4.64-4.60 (m, lH), 3.88 (dd, J = 12.6, 5.4 Hz, lH), 3.74-3.65 (m, lH), 3.60-3.38 (m, 2H).
Trans 4- t 4 - ~ 2 -Hydroaycarbonyl- 6-hydroAybenzoyl ) -3, 5-diL~oa~anzoyloAy]-3- (~-h2~roA~enz~i~o) -1- ~phenylsulfonyl) -pyrrolidine ~COMP0~1ND 562 ) To a solution of the previous product (183 mg, 0.164 mmol) in THF (7.4 ml) and ethanol (7.4 ml) was added Pd(OH)2 (92 mg. of a 20% by weight powder). The flask was evacuated and filled with H2 twice, then stirred under H2 (1 atm) for 20 h. The suspension was filtered through Celite, washed through with methanol (50 ml), and evaporated to a yellow oil.
Purification by HPLC (21 x 250 mm Cl8 column) provided the title compound (75 mg, 69%) as a fluffy yellow powder after ~09412W62 21~ 7 412 PCT~S94/022~
lyophilization: mp 185-208C; IR (KBr) 3402, 1709, 1636, 1608, 1232 cm ; H NMR (CD30D) ~ 7.53 (d, J = 8.2 Hz, 2H), 7.49 (d, J = 8.7 Hz, 2H), 7.32 (d, J = 7.3 Hz, lH), 7.22-7.11 (m, 3H), 7.09 (dd, J = 8.1, 7.9 Hz, lH), 6.84 (d, J = 7.2 Hz, lH), 6.61 (d, J = 8.7 Hz, 2H), 6.35 (s, 2H), 4.99 (app t, J = 2.2 Hz, lH), 4.92 (dd, J = 5.6, 2.8 Hz, lH), 3.62 (dd, J = 13.0, 4.3 Hz, lH), 3.50 (dd, J = 10.8, 6.0 Hz, lH), 3.39 (dd, J = 10.7, 2.4 Hz, lH), 3.32 (bd, J = 13.1 Hz, lH); HRMS (M + H) calcd 663.6315, found 663.1302; Anal. Calcd- for C32H26N20l2S 1 H2O:
C, 56.47; H, 4.15; N, 4.12; S, 4.71; found: C, 56.56; H, 4.17;
N, 4.09; S, 4.58.
-094/20062 2 1 5 7 1 1 2 PCT~S94/0~
Tr~ns-1-(4-hy~-o~yLehz~mi~o)-2-l4-~2-hy~roxy-6-meth benzoyl)-3,5-dihydroxybenzoyloxyl]cycloheptane ~COMPOUND 566) HO OM
O O
H~ ~ H~ , K,CO,, HMPA ~ H~
b o ~ o Figure AD
A mixture of Compound 588 (55 mg, 0.1 mmol), iodomethane (0.05 ml. 0.8 mmol), and K2CO3 (28 mg, 0.2 mmol) in HMPA (0.2 ml) was stirred at 40-C for 1.5h, and the reaction was judged incomplete by TLC. Additional iodomethane (0.025 ml, 0.4 mmol) was added and stirring was continued for 2h. at 40 C. EtOAc (15 ml) was added and the resultant mixture was washed with H2O (3 x 10 ml) and brine (10 ml), dried (MgSO4), and evaporated. The residue was purified by preparative TLC
(SiO2, multi-elution with CH2C12:5% MeOH in EtOAc = 4:1) to give a yellow solid (34 mg, 61%). IR (KBr, cm1): 1712, 1634, 1607.
FBMS: M/Z = 564 (M + 1).
W094/20062 21~ 7 ~ 12 PCT~S94/02283 Trans-~-t~-~2-Hydroxycarbonyl-6-hydroxybenzoyl)-3,5-~ihydko~L~zoyloxy]-3-(4-hyd-~yL~z~mido)-l-(5-dimethyl~min l-naphthalene sulfonyl)perhydroazepine (COMPOUND 569) O HO~
~OH OH,CH3 ~OH OH
H~o~ H ~ Cl ~I TEA ~ HHo~ Q~
,~ O N~ +~ S~ MeOH ~ ~O~N~ CH3 CF,CO2H NH CF,C02H \~ N~ SJ3~ ~ C~
569 Oo Racemic balanol (preparation described in Compound 508; 100 mg, 147 ~mol) was dissolved in methanol (1 ml) and treated with triethylamine (204 ~1, 1.47 ~mol) and dansyl chloride (39.5 mg, 146.5 ~mol) in methylene chloride (1 ml).
After stirring at room temperature for 3 h, the mixture was concentrated under vacuum to a yellow film. The residue was dissolved in DMF (2 ml) and chromatographed on a Dynamax-60 C18 column (41 mm ID x 25 cm length) using a linear gradient from 100% A (0.1% TFA and 5% acetonitrile in water) to 100% B (pure acetonitrile) over 60 m at 25 ml/min. The clean product, which eluted in 40 m, was freeze-dried to give a yellow powder (33 mg, 29%): m.p. 185-187-C dec; 1H-NMR (DMSO, 300 MHz) 1.73-2.12 (4H, m), 2.85 (6H, s), 4.30-4.41 (lH, m), 5.20 (lH, pseudo t), 6.75-6.87 (4H, m), 7.07 (lH, d, J = 8 Hz), 7.30 (2H, t), 7.39 (lH, d, J = 8 Hz), 7.62-7.70 (4H, m), 8.06 (lH, d, J
= 8 Hz), 8.24 (lH, d, J = 9 Hz), 8.50 (lH, d, J = 9 Hz), 9.91 (lH, 8), 11.68 (lH, s); IR (KBr): cm1 3399, 2361, 2340, 1702, 1677, 1635, 1607, 1542, 1507, 1462, 1425, 1371, 1319, 1283, 1237, 1200, 1139, 1103, 1064, 991, 920, 848, 793, 767, 723, 669, 581, 542, 532. Anal. Calcd. for C~oH37N3O12S 2H2O .92TFA
W094/20062 215 7 ~12 PCT~S94/022~
.15CH3CN: C, 54.37; H, 4.58; N, 4.73; S, 3.44. Found: C, 57.35; H, 4.51; N, 4.63; S, 3.17. LRMS (FAB) m/z 784.0 (783.2 calcd for C40H37N3O12S).
21~741~
wos~/20062 PCT~S94/02283 Tr~ns- ~ - t ~- ~ 2 -Carboxy-6-hydroxy~enzoyl ) -3, 5-dihydroxybenzoyloxy] -3- ~ 4 -hydroxy~enz~mido ) -1- ~ 2 -nitrobenzenesulfonyl)perhydroazepine, trifluoracetic acid salt COMPOIIND 57 3 ) o HO~=
~OH OH ~OH OH
H~o~ H~ Cl~ ~ TEA , H
~ o s ~ MeOH ~ O H~
CF,CO2H NH
CF,CO2H S~
Racemic balanol (preparation described in Compound 508; 100 mg, 147 ~mol) was dissolved in methanol (1 ml) and treated with triethylamine (204 ~1, 1.47 ~mol) and 2-nitrobenzene sulfonyl chloride (48.7 mg, 219.7 ~mol) in methylene chloride (1 ml). After stirring at room temperature for 3 h, the mixture was concentrated under vacuum to a yellow film. The residue was dissolved in DMF (2 ml) and chromatographed on a Dynamax-60 C18 column (41 mm ID x 25 cm length) using a linear gradient from 100% A (0.1% TFA and 5%
acetonitrile in water) to 100% B (pure acetonitrile) over 60 m at 25 ml/min. The clean product, which eluted in 33 m, was freeze-dried to give a yellow powder (28 mg, 26%): m.p.
158-160-C dec; lH-NMR (DMSO, 300 MHz) ~ 1.81-1.97 (3H, m), 2.06-2.17 (lH, m), 4.43-4.57 (lH, m), 5.18-5.28 (lH, m), 6.75 - (2H, s), 7.04 (lH, d, J = 8 Hz), 7.22-7.31 (3H, m), 7.36 (lH, d, J = 8 Hz), 8.05 (lH, t), 8.20 (lH, d, J = 8 Hz), 8.20 (lH, d, J = 8 Hz), 8.85 (lH, d, J = 8 Hz), 9.87 (lH, s), 11.67 (lH, s); IR (KBr): cm 3430, 3412, 1701, 1676, 1636, 1604, 1545, W094/20062 PCT~S94/02283 21~7~1~
1496, 1425, 1370, 1290, 1237, 1201, 1148, 1104, 1063, 1016, 993, 921, 874, 856, 799, 761, 738, 724, 586. Anal. Calcd. for C34H2~N30l4S 1-2 H20 1.1 TFA: C, 49.25; H, 3.71; N, 4.76; S, 3.63. Found: C, 49.22; H, 3.70; N, 4.72; S, 3.39. LRMS (FAB) m/z 735.9 (735.68 calcd for C34H29N3014S).
w094/20062 213 ~ ~ ~ 2 PCT/US94/0~
Tran8-4- ~- (2-Carboxy-6-hy~roxybenzoyl)-3,5-dihydroxybenzoyloxy]-3- (~-hydroxybenzamido) -1- ~
nitrobenzenesulfonyl)perhy~roazepine, trifluoracetic acid salt COMPO~ND 57 ~ ) o HO~
~OH OH ~=~OH OH
~~ H~ Cl` ~ TEA ~ HHo O .S ~ MeOH~ H~
~ ~ O O O ,~ j~ ,~,, NO2 CF~Co2H \~, NH CF~Co2H \ N` S
574 Oo Figure AG
Racemic balanol (preparation described in Compound 508; 100 mg, 147 ~mol) was dissolved in methanol (1 ml) and treated with triethyiamine (204 ~1, 1.47 ~mol) and 4-nitrobenzene sulfonyl chloride (48.7 mg, 219.7 ~mol) in methylene chloride (1 ml). After stirring at room temperature for 3 h, the mixture was concentrated under vacuum to a yellow film. The residue was dissolved in DMF (2 ml) and chromatographed on a Dynamax-60 C18 column (41 mm ID x 25 cm length) using a linear gradient from 100% A (0.1% TFA and 5%
acetonitrile in water) to 100% B (pure acetonitrile) over 60 m at 25 ml/min. The clean product, which eluted in 34 m, was freeze-dried to give a yellow powder (12 mg, 11%): m.p.
186-188-C dec; lH-NMR (DMSO, 300 MHz) ~ 1.80-1.97 (3H, m), 2.07-2.18 (lH, m), 3.13 (2H, pseudo t), 4.42-4.53 (lH, m), 5.17-5.28 (lH, m), 6.75 (2H, d, J = 6 Hz), 7.03 (lH, d, J = 8 Hz), 7.19 (lH, d, J = 9 Hz), 7.26 (lH, t), 7.34 (lH, d, J = 8 Hz), 7.63 (lH, d, J = 8 Hz), 7.76 (lH, d, J = 9 Hz); 7.88 (lH, d, J = 9 Hz), 8.15 (2H, d, J = 9 Hz), 8.43 (2H, d, J = 9 Hz), W094/20062 ~15 7 41 ~ PCT~S94/022~
~ - 276 -9.84 (lH, pseudo s, 11.71 (lH, pseudo s); IR (KBr): cm 3422, 3273, 3250, 3108, 3081, 2873, 2361, 2339, 1676, 1636, 1606, 1536, 1497, 1426, 1369, 1288, 1232, 1200, 1148, 1093, 1072, 1013, 960, 920, 874, 856, 761, 724, 681, 606, 568. Anal.
Calcd. for C34H29N3Ol4S 1 H2O 1.2 TFA .17 CH3CN: C, 49.17;
H, 3.67: N, 4.94; S, 3.57. Found: C, 49.16; H, 3.57; N, 4.64;
S, 3.36. LRMS (FAB) m/z 736.1 (735.68 calculated for C34HZ9N3ol4s ) -W094/20062 2 1 ~ 7 4 1 2 PCT~S94/02283 Tr~ns-3-(4-hy~roxybenz~mido)-~-t~-~2-hy~roxy-6-(2-methyl-propyloxy)c~rbonylhen~oyl)-3,5-~ihy~roxybenzoyloxy]pyrroli~ine trifluoro~cetic ~cid s~lt (COMPOUND 577) BnO~n1, (COCI)2, DMF, CH2C12 BnO~O o o~n OBn OFB~ HO", NJb~ ~, HJ~3 BOC BOC
DMAP, EtJN
H2, Pd(OH)~ / C
TFA
HO E~OH / EtOAc ~N)~ $N~ QH
BOC
TFA
~0 ~ O OH
H~
TFA
21S741`~
~ ~ - 278 -Trans-N-t-butoxycarbonyl-3-(~-benzyloxybenzam~ ~o)-~-t~-(2-benzyloxy-6-benzyloxycarbonylbenzoyl)-3,5-dibenzyloxy]-pyrrolidine 4-[4-(2-benzyloxy-6-benzyloxycarbonylbenzoyl)]-3,5-dibenzyloxybenzoic acid (1.47 mmol, 996 mg) and 15 ml anhydrous CH2Cl2 in a dry round-bottom flask were cooled in an ice/water bath under N2. To this was added oxalyl chloride (2.87 mmol, 0.25 ml) and 5 drops of DMF. This was allowed to stir for 2 hours while the bath melted. TLC (2:1 hexanes:EtOAc) indicated complete formation of the acid chloride. The solvent was removed in vacuo.
In a 200 ml dry round-bottom flask was added trans-N-t-butoxycarbonyl-3-(4-benzyloxybenzamido)-4-hydroxypyrrolidine (1.26 mmol, 500 mg) in 12 ml anhydrous CH2C12 under N2. To this was added triethylamine (3.6 mmol, 0.5 ml) and DMAP (150 mg).
A solution of the acid chloride generated above in 10 ml anhydrous CH2Cl2 was added via cannula. This was allowed to stir under N2 at room temperature overnight. The reaction mixture was then diluted with CH2C12, washed with sat. NaHCO3, brine, then dried over MgSO" and concentrated in vacuo. The crude product was purified via flash column chromatography using 5% acetone/CH2Cl2 as the eluent. Trans-N-t-butoxycarbonyl-3-(4-benzyloxybenzamido)-4-[4-(2-benzyloxy-6-benzyloxycarbonylbenzoyl) -3,5-dibenzyloxy benzoyloxy~
pyrrolidine (1.08 mmol, 1.15 g) was obtained in 8696 yield.
Trans-N-t-butoxyoarbonyl-3-(~,-hy~roxybenzamido)-~-[~-(2-hy~roxy-6-¢~,bG.~benzoyl)-3,5-dihydroxybenzoyloxy]pyrrol~dine To a 500 ml 3-neck round-bottom flask was added trans-N-t-butoxycarbonyl-3-(4-benzyloxybenzamido)-4-[4-(2-benzyloxy-6-benzyloxycarbonylbenzoyl) -3, 5-dibenzyloxy benzoyloxy]pyrrolidine (1.02 mmol, 1.08 g) in 17 ml EtOAc and 70 ml ethanol under N2. To this was added trifluoroacetic acid (2.55 mmol, 0.20 ml) and Pd(OH)2/C (730 mg) followed immediately by introduction of H2 at 1 atmosphere. After a reaction time of 3.5 hours, the reaction was flushed with N2 -~094/20062 21~ 7 ~ ~ 2 PCT/US94/02283 and filtered through Celite, rinsing with ethanol. Following concentration in vacuo, crude product (644 mg) was obtained in quantitative yield. A small portion was purified via HPLC (21 x 250 mm C18 column, gradient ~B = 0 to 50 over 60 min. where A = 0.1~ TFA, 5% CH3CN in water and B = CH3CN, 15 ml/min. W =
254 nm) for characterization and the remainder was used crude in subsequent reactions. m.p. 196C(dec). IR (KBr) 3375(br), 2978, 1704, 1660, 1637, 1607, 1506, 1426, 1368, 1231 cm . H
NMR CD30D, ~ 8.52 (d, lH), 7.72 (d, 2H), 7.49 (d, lH), 7.26 (t, lH), 7.01 (d, lH), 6.91 (s, 2H), 6.80 (d, 2H), 5.40 (m, lH), 4.63 (m, lH), 3.87 (m, 2H), 3.50 (m, 2H), 1.47 (s, 9H). LRMS
(M + 1) cacld for C31H31N2O12 623.2, found 623.2. Anal. Calcd for C3lH3lN2O12 1-5 H2O: C, 57.317; H, 5.120; N, 4.312. Found: C, 57.26; H, 5.18; N, 4.47.
Trsns-N-t-butoxyc~rbonyl-3-(~-hydroxybenz~mi~o)-~-t4-~2-hydroxy-6-~2-methylpropyloxy)¢~rbonylbenzoyl)-3,5-dihydroxybenzoyloxy]pyrrolid~ne (COMPOUND 6~6) To a round-bottom flask was added trans-N-t-butoxycarbonyl-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-carboxylbenzoyl)-3,5-dihydroxybenzoyloxy]pyrrolidine (0.lS
mmol, 95 mg) in 5 ml DMF. To this was added NaHCO3 (0.23 mmol, 19 mg) and 1-iodo-2-methylpropane (0.75 mmol, 0.09 ml). This was allowed to stir under N2 for 7 days with additional 1-iodo-2-methylpropane being added each day. A total of 48 additional equivalents (7.2 mmol, 0.83 ml) were added over the reaction period. The reaction mixture was diluted with EtOAc and washed with water 3 times. The aqueous layer was back extracted with EtOAc and the organic layers combined and dried over MgSO4 then concentrated in vacuo. The crude product was purified via HPLC
(21 x 250 mm C18 column, gradient %B = 25 to 100 over 60 min.
where A = 0.1% TFA, and 5% CH3CN in water, B = CH3CN, 15 ml/min.
W = 254 nm) to isolate trans-N-t-butoxycarbonyl-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-(2-methylpropyloxy)-carbonylbenzoyl)-3,5-dihydroxybenzoyloxy]pyrrolidine (46.5mg, 44% yield).
WO 9~/20062 215 7 4 ~ 2 PCT/US94/02283 Trans-3- ~4-hydroxybenzami~o) -4- t4- ~2-hydroxy-6- ~2-methylpropyloxy)carbonylbenzoyl)-3,5-dihy~roxybenzoyloxy]-pyrrolidine trifluoroacetic acid ~alt ~COMPOlrND 577) Trans-N-t-butoxycarbonyl-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6- (methylpropyloxy) carbonylbenzoyl) -3, 5-dihydroxybenzoyloxy]pyrrolidine (46.5 mg, 0.068 mmol) was dissolved in 0.75 ml neat trifluoroacetic acid and allowed to stir at room temperature under N2 for 45 minutes at which time TLC (75% CH2C12, 24% MeOH, 1% (10% aq.) NH40H) indicated the reaction was complete. This was diluted with toluene and concentrated in vacuo to yield 42.7 mg (91% yield) of crude product. Purification via HPLC (21 x 250 mm C18 column, gradient %B = 0 to 100 over 60 min. where A = 0.1% TFA, 5%
CH3CN in water, B = CH3CN, 15 ml/min. W = 254 nm) yielded trans-3- (4-hydroxybenzamido) -4- [4- (2-hydroxy-6- (2-methylpropyloxy ) -carbony1benz oy1 ) - 3, 5 -dihydroxybenzoyloxy~pyrrolidine trifluoroacetic acid salt (Compound 577, 32 mg, 6896 yield) as a yellow solid. m.p.
150-156-C (dec.). IR (KBr) 3191 (br), 2968, 1677, 1607, 1508, 1426, 1373, 1293, 1201 cm-l. lH NMR, DMSO-d6, ~, 11.74 (s, 2H), 10.13 (s, lH), 10.03 (s, lH), 9.20 (br, NH), 8.51 (d, lH), 7.74 (d, 2H), 7.44 (d, lH), 7.35 (t, lH), 7.12 (d, lH), 6.95 (s, 2H), 6.84 (d, 2H), 5.50 (m, lH), 4.60 (m, lH), 3.88 (d, 2H), 3.72 (m, 2H), 3.52 (m, 2H), 1.80 (q, lH), 0.83 (d, 6H).
LRMS (M + 1) calcd for C30H3lN2Olo 579.20, found 579.1. Anal.
calcd for C30H30N2Olo C2HF3O2 1.3 H2O: C, 53-68; H~ 4-73; N~
3.91. Found: C, 53.78; H, 4.54; N, 3.99.
~o9~l2oo62 ~1574~2 Tr~n8~ -hy~roxybenz~mi~o) -2- t~- (2-hy~roxy-6-hy~roxyc~rbonylbenzoyl)-3,5-~ihy~roxybenzoyloxyl]cyclohept~ne (COMPO~ND 588) C~ N-~CO~ CH~ (9 I~OH ~O (~ N~ TBDI~tS Cl OBn OTB0MS H?~Pd-C OTBDMS . ' H ~) N~ M OH (~ BnO~ CO~H, THf TBDMS~N~
CO~Bn OBn OlloO Olln g~n O n Bu,NF THF H~j o EI,N,DMAP,CH2CI, BnO O N~
~J ~ O
a~,H
H?~Pd(OH)~ C ~H ~ H
M~OH THF o ~
21~7~12 ~ ; - 282 -NaOAc (426 mg, 5.2 mmol) was dissolved in a solution of peroxyacetic acid in acetic acid (32 wt. %, 21.9 ml, 104 mmol) and the resultant solution was added dropwise over 30 min. to a mixture of cycloheptene (10 g, 104 mmol) and Na2CO3 (44.1 g, 416 mmol) in CH2C12 (100 ml) at 5 C. The mixture was allowed to stir at room temperature for 3h with occasional cooling using a water bath. All solid material was removed by filtration, and the filtrate was distilled at atmospheric pressure using a vigreux column to remove most of the CH2Cl2, giving a colorless liquid which was shown by Hl NMR to be a mixture of the epoxide and CH2C12. The purity was estimated to be ca. 73% in the epoxide (total 15.93 g, 100% crude yield).
This material was used without further purification in the next step.
A mixture of the epoxide from the previous step (10 g, 65 mmol), NaN3 (27.5 g, 423 mmol), NH4Cl (10.44 g, 195 mmol), MeOH (162 ml), and H2O (20 ml) was stirred at reflux for 24h.
The solid was removed by filtration and the filtrate was evaporated. The residue was treated with 0.5 N aq. NaOH (50 ml) and extracted with CH2C12 (3 x 30 ml). The combined CH2Cl2 extracts were washed with H2O (50 ml), brine (50 ml), dried (MgSO4), and evaporated to give the azide (8.39 g, 83%).
The azide (7 g, 45 mmol) in N,N-dimethylformamide (10 ml) was added to a solution of t-butyldimethylsilylchloride (6.8 g, 45 mmol) and imidazole (3.07 g, 45 mmol) in N,N-dimethylformamide (35 ml). The resultant mixture was stirred at room temperature for 16h, diluted with Et2O (70 ml), washed with H2O (4 x 30 ml) and brine (50 ml), dried (MgSO4), and evaporated to give a pale yellow oil (11.33 g, 93%).
A mixture of the product of the previous reaction (8.1 g, 29.95 mmol) and 5% Pd on carbon (1.59 g, 2.5 mol. %) in MeOH
was stirred vigorously under 1 atm H2 at room temperature for 20h. The catalyst was removed by filtration over Celite and the filtrate was evaporated. The residue was purified by flash chromatography (SiO2, Et2O: Hexane = 2:1) to give the amine as a colorless oil (5.27 g, 72~).
~094/20062 213 7 ~1~ PCT/US94/02283 A mixture of 4-benzyloxybenzoic acid (3.287 gj 14.40 mmol) and 1,1'-carbonyldiimidazole (2.335 g, 14.40 mmol) in THF
was stirred at room temperature for 2h. To the resultant slurry was added a solution of the amine product of the previous reaction (3.07 g, 12.56 mmol) in THF and stirring was continued at room temperature for 24h. The mixture was diluted with CH2Cl2 (40 ml), washed with H20 (3 x 15 ml) and brine (2 x 15 ml), dried (MgS04), and evaporated. The residue was purified by flash chromatography (SiO2, Et20 : hexane = 1:5) to give the amide as a white powder (4.69 g, 82%).
Tetrabutylammonium fluoride in THF (lM, 10 ml, 10 mmol) was added to a solution of the amide product from the previous reaction (3.8 g, 8.36 mmol) in THF and the resultant yellow solution was stirred at room temperature for 2h. The mixture was poured into CH2C12 (150 ml), washed with H20 (3 x 30 ml) and brine (2 x 30 ml), dried (MgS04), and concentrated to about 30 ml. The precipitate was collected by filtration and washed with CH2Cl2 (3 x 3 ml). The combined filtrate and washes were concentrated to about 10 ml, and the resultant precipitate was collected, washed with CH2C12 (2 x 2 ml), combined with the first crop and dried under vacuum. This gave a white solid (2.24 g, 79%). The mother liquors were combined and chromatographed (SiO2, Et20 : hexane = 1:5 followed by CH2Cl2 to give an additional 256 mg of the product, total yield: 89%.
To a stirred solution of 4-(2-benzyloxycarbonyl-6-benzyloxy)benzoyl-3,5-dibenzyloxybenzoylchloride (350 mg, 0.52 mmol) in CH2Cl2 (1.5 ml) was added oxalyl chloride (0.067 ml, 0.77 mmol) and one drop of N,N-dimethylformamide. The resultant solution was stirred at room temperature for 2h and then evaporated to dryness. The residue was dissolved in CH2Cl2 (1 ml) and added to a mixture of the product of the preceding reaction (177 mg, 0.52 mmol) and Et3N (105 mg, 1.04 mmol) in CH2Cl2 (2 ml). The mixture was stirred at room temperature for 17h, diluted with CH2Cl2 (15 ml), washed with H20 (3 x 10 ml), dried (MgS04), and evaporated. The residue was chromatographed wos4/20062 215 7 ~12 PCT~S94/02283 (sio2~ Et2O: hexane = 1:1, followed by Et2O: hexane: CH2Cl2 =
1:1:1) to give a white solid (166 mg, 32%).
Pd(OH)2 on carbon (20 wt. % contains > 50% moisture;
22 mg, 0.016 mmol) and MeOH (1.6 ml) were added to a solution of the product of the prec~ing reaction (160 mg, 0.16 mmol) in THF (1.6 ml) and the resultant mixture was stirred under 1 atm H2 at room temperature for 17h. The catalyst was removed by filtration over Celite and the Celite pad was washed with MeOH
(15 ml). The filtrate and washes were combined and evaporated to give a yellow solid (83 mg, 95%) (COMPOUND 588). m.p. 186-C
(dec). Anal. calcd for C29H2,NOlo-lH2O: C, 61.37; H, 5.15; N, 2.47. Found: C, 61.41; H, 5.29; N, 2.36.
2 1 ~ 741~
Trans-3- (~-hy~lohyLt:nz~lmido) -4- t4- ~2-hy~roxy-6-hy~ro2ycarbonyl benzoyl) -3,5-~ihy~roxybenzoyloxyl]pyrroli0inium trifluoroacetate (COMPO~ND 589) ~nd 1-}~exa~ecanoyl-trzms-3-~hy~GhyL ~nzami~o) -~.- t 40 ~2-hy~roxy-6-hy~roxycarbonylbenzoyl ) -3, 5-~ihy0roxybenzoyloxyl ] pyrrol i~ine ~ COMPOIJND S 9 0 ) co2~
e~~i C~ ~n ~H~en EloN~ DMAP, CH,C~I
o~L OEln O 06n CO~H
~ ~ c~(c~"coa MeOH, THF O ~ O
CF,CO2H
C~2H
~,~OH D~ H
HO~ ~H~J
,~0,~ 0 0~ (CH~"C~
To a stirred solution of 4(2-benzyloxycarbonyl-6-benzyloxy)benzyl-3,5-dibenzyloxybenzoylchloride (350 mg, 0.52 mmol) in CH2Cl2 (1.5 ml) was added oxalyl chloride (0.067 ml, ~og~/20062 215 7 412 PCT/US94/02283 0.77 mmol) and one drop of N,N-dimethylformamide. The resultant solution was stirred at room temperature for 2h and then evaporated to dryness. The residue was dissolved in CH2Cl2 tl.5 ml) and added to a-mixture of trans-3-(4-hydroxy-1-benzyloxycarbonyl pyrrolidine (232 mg, 0.53 mmol; for preparation see Compound 585), Et3N (0.14 ml, 1.04 mmol), and 4-(N,N-dimethylamino)pyridine (6 mg, 0.052 mmol) in CH2Cl2 (1.5 ml). The mixture was stirred at room temperature for 17h, diluted with CH2Cl2 (15 ml), washed with H2O (3 x 10 ml), dried (MgSO4), and evaporated. The residue was chromatographed (SiO2, Et2O:hexane = 1:1, then EtzO:hexane:CH2Cl2 = 1:1:0.5) to give the ester (230 mg, 40%) (COMPOUND 652).
To a solution of Compound 652 (200 mg, 0.18 mmol) in THF (1.8 ml) were added MeOH (1.8 ml), Pd(OH)2 on carbon (20 wt.~, contains 2 50% moist; 25 mg, 0.018 mmol), and CF3CO2H (41 mg, 0.36 mmol). The mixture was stirred at room temperature under 1 atm H2 for 17h. The volatile components were removed by evaporation, and the residue was taken up in MeOH, filtered through Celite, and evaporated to give a yellow solid (88 mg, 72~) (COMPOUND 589).
Palmitoyl chloride (10 mg, 0.038 mmol) was added to a stirred solution of Compound 589 (24 mg, 0.038 mmol) in pyridine (0.4 ml). The mixture was stirred at room temperature for 16h and TLC showed that the reaction was incomplete. More palmitoyl chloride (5 mg) was added and stirring was continued for 16h. The reaction mixture was evaporated to remove pyridine leaving a yellow syrup with some solid material which was shown to contain starting material by lH NMR. This material was dissolved in pyridine (0.4 ml) and treated with palmitoyl chloride (15 mg). The mixture was stirred at room temperature for 24h, evaporated, and chromatographed (SiO2, EtOAc followed by S% HOAc in acetone) to give a yellow solid (19 mg, 66%) which was shown by lH NMR to be the desired amide (Compound 590) with some contamination. m.p. 175-178C (dec).
Anal. calcd for C42H52N2Oll-3H2O: C, 61.90; H, 7.17; N, 3.44.
Found: C, 61.85; H, 7.07; N, 3.62.
_~094/20062 - 287 - PCT~S94/02283 Tran~-3-~-hydL~L~z~mido)-4-[4-(2-hydroxy-n~phth-1-oyl)-3,5-dihydroxybenzyoyloxyl]pyrrolidine trifluoroacetic ~ci~ s~lt (COMPOUND 591) H~;;5 E~,N. O~P, C~
0~ OE~n OJ` Oan ~H 01 1 U O~, THF
N CF~
To a mixture of 4-(2-benzyloxy-naphth-1-oyl)-3,5-dibenzyloxylbenzoic acid (297 mg, 0.5 mmol) and 2 drops of N,N-dimethylformamide in CH2C12 (1.5 ml) was added oxalyl chloride (0.065 ml, 0.75 mmol) dropwise over 2 min. The resultant mixture was stirred at room temperature for 2h and the volatile components were evaporated. The remaining yellow solid was dried under vacuum for lh, dissolved in CH2Cl2 (1.5 ml), and added to a solution of trans-3-(4-hydroxybenzamido-4-hydroxy-1-benzyloxycarbonyl pyrrolidine (168 mg, 0.38 mmol; for preparation see Compound 585), Et3N (0.1 ml, 0.76 mmol), and 4-(N,N-dimethylamino)pyridine (5 mg, 0.038 mmol) in CH2C12 at 5C.
wos4noo62 215 ~ 4 ~- ~ PCT/US94/022~
The mixture was stirred at room temperature for 16h, diluted with CH2Cl2 (10 ml), washed with H2O (3 x 10 ml), dried (MgSO4), and evaporated. The residue was chromatographed (SiO2, Et2O:hexane = 1:1, followed by Et2O:hexane:CH2Cl2 = 1:1:0.5) to give a white solid (336 mg, 66%) (COMPOUND 653).
Pd(OH)2 on carbon (20 wt.%, contains > 50% moist; 21 mg, 0.015 mmol), followed by CF3CO2H (34 mg, 0.3 mmol) and MeOH
(1.5 ml), was added to a solution of Compound 653 (155 mg, 0.15 mmol) in THF (1.5 ml). The resultant mixture was stirred at room temperature under 1 atm H2 for 2Oh. The catalyst was removed by filtration over Celite and the Celite pad was washed with MeOH (20 ml). The combined filtrate and washes were evaporated to give a yellow solid (85 mg, 88%) (COMPOUND 591).
~ ~O9~/20062 2157~12 l-Isopropyl-tr~n~-3-(~-hydroxybenz~mi~o~ y~2~by ~ ~y~
hydroxyc~rbonylhe~7cyl)-3~5-dihy~roxybenzoyloxyl]pyrrolidinium trifluoro~cet~te (COMPOUND 592) OBn O~n ,~ ~,O~;
~S ~, ~ o ~=/ NBH~. ~H)--\ O E~,~. DMAP, CHICI~
H~, H CF,CO,H
~eO ~e o ~H~ H, Pd(OH~,-C ~o c~ TF~ 0~. T\~F O ~ o ~ Cf~
6~ 592 W09~i20062 215 7 ~ PCT/US94/02283 To a solution of trans-3-(4-benzyloxybenzamido)-3-hydroxy pyrrolidine (213 mg, 0.5 mmol) in acetic acid (2.5 ml) was added NaBH4 (95 mg, 2.5 mmol) in small portions. After the H2 evolution had ceased (ca. 10 min.) acetone (0.18 ml, 2.5 mmol) was added and the mixture was stirred at room temperature for 16h. The reaction mixture was basified with 2N KOH and the resultant cloudy mixture was extracted with CH2C12 (3 x 15 ml).
The combined CHzCl2 extracts were dried (MgSO4), and evaporated to give a colorless oil (163 mg, 92%).
Oxalyl chloride in CH2C12 (2 M, 0.39 ml, 0.78 mmol) was added dropwise to a solution of 4-(2-benzyloxy-6-benzyloxycarbonylbenzoyl)-3,5-dibenzyloxybenzoic acid (353 mg, 0.52 mmol) and a drop of DMF in CH2C12 (2 ml) at 5 C. The mixture was stirred at room temperature for 2h, then evaporated to remove the solvent and excess oxalyl chloride. The residue was dried in vacuo for lh, dissolved in CH2C12 (1 ml), and added to a mixture of the product of the preceding reaction (142 mg, 0.4 mmol), Et3N (81 mg, 0.8 mmol), and DMAP (6 mg, 0.054 mmol) in CH2Cl2 (2 ml) at 5-C. The mixture was stirred at room temperature for 17h, diluted with CH2C12 (15 ml), washed with H2O (3 x 10 ml), dried (MgSO4), and evaporated. The residue was chromatographed (SiO2, Et2O:CH2Cl2:hexane = 1:1:1 followed by Et2O:CH2Cl2:hexane = 2:2:1) to give a pale yellow oil (82 mg, 20%) (COMPOUND 654).
Pd(OH)2 on carbon(20 wt%, contains <50% moist, 11 mg, 0.008 mmol), trifluoroacetic acid (18 mg, 0.16 mmol), and MeOH
(1 ml) was added to a solution of Compound 654 (84 mg, 0.08 mmol) in EtOAc (1 ml) and the mixture was stirred under 1 atm H2 contained in a balloon at room temperature for 16h. The mixture was filtered through Celite and the Celite pad was washed with MeOH (5 ml). The combined filtrates were evaporated to give a yellow solid (32 mg, 59%) (COMPOUND 592).
IR (KBr, cm ):1705, 1676, 1636, 1607. FBMS: M/Z = 565 (M + 1).
2157~12 ~O g~/20062 - 291 - PCT/US94/02283 Trans-~- t~- (2-carboxy-6-hydroxybenzoyl) -3,5-~ihy~roxybenzoyloxy]-3- (4-hy~roxybenzami~o) -1-[ (phenylamino) carbonyl] -pyrroli~ine (COMPO~JND 593) NE~ OBn H-TFA O~N
H
C 02Bn iPr2NE~, ¦ OBn DMAP, ~3 c OC 1 OBn 08n H~OhN~ Pd(OHk ~ O OBn H02C~ooH N 2 ~-08n I Fh (~) -Trans-~-Hydroxy-3- (~-benzyloxybenz~mido)-l-tphenylamino)carbonyl]-pyrrolidine To a solution of (+) trans-4-hydroxy-3-(4-benzyloxybenzamido pyrrolidine TFA (100 mg, 0.235 mmol) in methanol (4.7 ml) were added triethyl amine (66 ~L, 0.470 mmol, 2.0 eq) then phenyl isocyanate (26 ~L, 0.235 mmol, 1.0 eq).
The cloudy mixture was stirred at room temperature under N2 45 min, then poured into H2O (30 ml) and extracted with CH2Cl2 (3 x 25 ml). The organic layers were combined, dried (MgSO4), filtered and evaporated to a white solid (82 mg, 81%): 1H NMR
(300 MNz, CD30D ~ 7.60 (d, J = 8.8 Hz, 2H), 7.00-7.25 (m, llH), 6.85 (d, J = 8.9 Hz, 2H), 6.79 (d, J = 7.1 Hz, lH), 4.93 (s, 2H), 4.20-4.25 (m, lH), 4.16 (dd, J = 8.2, 3.1 Hz, lH), 3.73 W094/20062 21 S 7 ~12 ~ ~ PCT~S94/02283 (dd, J c 11, 6.4 Hz, lH), 3.57 (dd, J = 11.4, 5.1 Hz, lH), 3.12 (dd, J = 11.0, 3.5 ~z, lH), 3.26 (dd, J = 11.1, 2.8 Hz, lH).
( + ) -Tran~-~- t ~ - ~ 6-benzyloxy-2- ( benzyloxycarbonyl ) benzoyl ] - 3, 5-~ibenzyloxybenzoyloxy) -3- (~.-benzyloxybenzamido) -1-(phenylamino) c~rbonyl]-pyrrolidine (CON~uNv 655) To a solution of the product of the preceding reaction (80 mg, 0.19 mmol), 4-dimethylaminopyridine (26 mg, 0.21 mmol, 1.1 eq) and diisopropylethylamine (37 ~L, 0.21 mmol, 1.1 eq) in CH2Cl2 (3.2 ml) was added a solution of acid chloride (0.21 mmol) in CH2C12 (1.6 ml). The mixture was stirred at room temperature under N2 18 h, then poured into 5% HCl (30 ml) and extracted with CH2Cl2 (3 x 25 ml). The organic layers were combined, dried (MgSO4) filtered and evaporated. Flash column chromatography of the golden residue (1.1 hexanes:ethyl acetate) on silica gel provided the title product (110 mg, 53%) as an off white foam: 1H NMR (300 MHz, CDCl3) ~ 7.89 (d, J = 8.8 Hz, 2H), 6.9-7.6 (m, 26H), 6.82 (d, J = 7.5 Hz, 2H), 5.4-5.5 (m, lH), 5.12 (s, 2H), 5.02 (s, 2H), 4.80 !m, lH), 4.76 (s, 4H), 4.68 (s, 2H), 4.0-4.05 (m, lH), 3.95-4.0 (m, lH), 3.65-3.7 (m, lH), 3.6-3.65 (m, lH).
Tr~ns- ~ - t 4 - ~ 2 -C~rboxy- 6-hydroxybenzoyl ) -3, 5-dihydroxybenzoyloxy] -3- (4-hy~roxybenzami~o) -~-~phenylamino) carbonyl]-pyrrolidine ~COMr~u~v 593) To a round bottom flask containing Compound 655 (110 mg, 0.101 mmol) were added Pd(OH)2 (22 mg of a 20% powder) then THF (2.2 ml) and ethanol (2.2 ml). The flask was evacuated and filled with H2 twice, then allowed to stir under H2 (1 atm) for 23 h. The suspension was filtered, washed through with methanol (40 ml) and evaporated to a yellow solid.
Purification by reverse phase HPLC (618 column) provided the title product as a yellow powder after lyophilization (24.9 mg, 39%): m.p. 183-208 (dec); IR (KBr) 3458, 3336, 1719, 1679, 1622, 1227, 760 cm ; lH NMR (300 MHz, CD30D) ~ 7.53 (d, J = 8.6 Hz, 2H), 7.28 (d, J = 7.7 Hz, lH), 7.20 (d, J = 7,7 Hz, 2H), 7.0-7.1 (m, 3H), 6.82 (d, J = 8.3 Hz, 2H), 6.73 (s, 2H), 6.62 215741~ ~
(8.7, J = d Hz, 2H), 5.25-5.35 (m, lH), 4.5-4.6 (m, lH), 3.75-3.9 (m, 2H), 3.4-3.5 (m, 2H): MS m/e calc'd for C33H2~N3Oll:
642.1732, found 642.1865; Analysis calc'd for C33H27N3O1l-0.5 TFA-1.25 H2O: C,56.63; H, 4.19; N, 5.83; found: C, 56.85; H, ~4.26; N, 5.96.
W094l20062 ~lS 7 4 1 2 - 294 - PCT~S94/02283 ~)Trans-4t4-(2-Carboxy-6-hy~roxybenzoyl)-3,5-dihydroxybenzoyloxy]-3-(4-hydroxybenzamido)-1-t(methylamino)c~rbonyl]-pyrroli~ine (COMPOUND S9~) HO~p~ , NE~3 ~ ~3`08n H-lFA H
C02Bn Pr2NEt, ¦ OB n DMAP, ~'COCI
OBn OBn H 0~0 ~ EdlOHHkHF _~
~o h OH Oq~ ~j OBn H02C~b~ o~N,CH3 BnO2C~_gBn N
(~)-Trans-4-Hydroxy-3-(~-benzyloxybenzamido)-1-t(methylsmino)c~rbonyl]-pyrrolidine To a solution of (+)-trans-4-hydroxy-3-(4-benzyloxybenzamido)pyrrolidine TFA (100 mg, 0.235 mmol) in methanol (4.7 ml) were added triethyl amine (66 ~1, 0.470 mmol, 2.0 eq) then methyl isocyanate (14 ~L, 0.235 mmol, 1.0 eq). The cloudy mixture was stirred at room temperature under N2 1.5 h, then more methyl isocyanate (14 ~L, 1.0 eq) was added. The mixture was allowed to stir 10 min more, then poured into H20 (30 ml) and extracted with CH2C12 (3 x 25 ml). The organic layers were combined, dried (MgS04) filtered and evaporated to ~ W094/20062 21~ 7 412 PCT/US94/02283 a white solid (83 mg, 95%): H NMR (300 MHz, CD30D) ~ 7.60 (d, J = 8.8 Hz, 2H), 7.1-7.25 (m, 5M), 6.85 (d, J = 8.8 Hz, 2H), 4.66 (s, 2H), 4.15 (dd, J = 6.5, 3.4 Hz, lH), 4.10 (dd, J =
5.0, 3.4 Hz, lH), 3.57 (dd, J = 10.8, 6.5 Hz, lH), 3.43 (dd, J
= 11.1, 5.2 Hz, lH), 3.1-3.2 (m, 2H), 2.53 (s, 3H).
(+)-Trans-~-t~-~6-benzyloxy-2-~benzyloxycarbonyl)benzoyl]-3,5-dibenzyloxybenzoyloxy) -3- ~-benzyloxybenzamido) -1-[ ~methylamino) c~rhonyl~ -pyrrolidine ~COMPOUND CS6) To a solution of the product of the prececeding reaction (85 mg, 0.23 mmol), 4-dimethylaminopyridine (28 mg, 0.23 mmol, 1.1 eq) and diisopropylethylamine (40 ~L, 0.23 mmol, 1.1 eq) in CH2C12 (3.9 ml) was added a solution of acid chloride (0.25 mmol) in CH2C12 (1.9 ml). The mixture was stirred at room temperature under N2 16 h, then poured into 5% HCl (30 ml) and extracted with CH2C12 (3 x 25 ml). The organic layers were combined, dried (MgS04) filtered and evaporated. Flash column chromatography of the golden residue (98:2 CH2Cl2:methanol) on silica gel provided the title product (159 mg, 67%) as an off white solid: lH NMR (300 MHz, CDCl3) ~ 7.84 (d, J = 8.7 Hz, 2H), 7.3-7.4 (m, 5H), 7.1-7.3 (m, 16H), 7.06-7.08 (m, 6H), 6.9-7.0 (m, 3H), 6.84 (d, J = 7.6 Hz, 2H), 5.49 (m, lH), 5.14 (s, 2H), 5.07 (s, 2H), 4.78 (s, 4H), 4.70 (m, lH), 4.70 (s, 2H), 4.40 (m, lH), 3.95-4.0 (m, 2H), 3.53-3.60 (m, 2H), 2.80 (d, J = 4.5 Hz, 3H).
~ + ) -Trans- ~ - t ~ - ~ 2 -Carboxy- 6 -hydroxybenzoyl ) - 3, 5 -dihy~ro~cyenzoyloxy] -3- ~.-hy~roxybenzamido) -1-~methyl~ino) carbonyl]-pyrrolidine ~COMPOIJND 594) To a round bottom flask containing Compound 656 (159 mg, 0.154 mmol) were added Pd(OH)2 (40 mg of a 20% powder) then THF (3.0 ml) and ethanol (3.0 ml). The flask was evacuated and filled with H2 twice, then allowed to stir under H2 (1 atm) for 21 h. The suspension was filtered, washed through with methanol (50 ml) and evaporated to a yellow solid.
Purification by reverse phase HPLC (C18 column) provided the title product as a yellow powder after lyophilization (68.7 mg, ~VO 94/20062 21~ 7 412 PCT/US94/02283 77%): m.p. 178-198 (dec); IR (KBr) 3385, 1714, 1605, 1236, 763 cm; H NMR (300 MHz, CD30D) ~ 7.53 (d, J = 8.7 Hz, 2H), 7.30 (d, J = 7,7 Hz, lH), 7.06 (dd, J = 8.1, 7.9 Hz, lH), 6.80 (d, J = 8.3 Hz, lH), 6.70 (s, 2H), 6.62 (d, J = 8.7 Hz, 2H), 5.2-5.3 (m, lH), 4.4-4.5 (m, lH), 3.6-3.8(6 line mult, 2H), 3.2-3.4 (m, 2H), 2.53 (s, 3H); MS m/e calc'd for C28H25N3011: 580-1567~
found 580.1481; Analysis calc'd for C2~H25N3O11-0.3TFA 0.8H20:
C, 54.32; H, 3.96; N, 6.72; found: C, 54.61; H, 4.32; N, 6.68.
~O9~/20062 21 r 7 ~
- 297 - ~ PCT~S94/02283 ~ tr~ns-3-~3,~-~ihy~roxybenz~mido)-~-t~-(2-c~rboxy-6-hy~roxybenzoyl)-3,5-dihy~roxy]benzoyloxypyrroli~ine trifluoro~cetic ~cid ~lt (COMPOUND 675) BnO
-- ~ O Jonesox.
BnO ~ ~
~-J H 93%
OBn H NH2 ~ OBn - BnO H _ + Bn ~ O NaOH,CH2C12 HO\ NL_ CB OH 99% N
Z B
z BnO
O 1.(COCI)2cat.DMF,~ OBn OBn OBnCH2CI2 ~ ~
2. Et3N, DMAP,BnLOnO ~ H ~ OBn B~sOno~~OH CH2CI2.70%
O N
/ B
HO / 2. Pd(H)2 ,~--o / EtOAc-EtOH, TFA
OH OH
H~ ~ H ~ OH
O ~ N ~
WO9~/20062 21~ 7 ~1 2 PCT/US94102t83 Jones reagent (6ml) was added to a solution of 3,4-dibenzyloxybenzaldehyde (Aldrich, 1.0 g, 3.14 mmol) in acetone (20 ml) until the reaction remained the color of Jones reagent. The excess of Jones reagent was destroyed by adding i-PrOH and acetone was removed in vacuo. The slurry residue was taken into EtOAc, washed with brine, dried with Na2SO4, and concentrated to afford off-white solids (0.98 g, 93~).
To a suspension of 3,4-dibenzyloxybenzoic acid (0.467 mg, 1.40 mmol) in anhydrous CH2C12 (5 ml) was added cat. DMF and oxalyl chloride (2.0 M solution in CH2C12, 1.92 ml, 3.85 mmol) at room temperature The mixture was kept for stirring at room temperature for 2 hr. Solvents were removed and the acid chloride residue was taken into CH2C12 (5 ml) after drying over the vacuum for lhr. To a biphasic reaction mixture of N-CBZ-3-amino-4-hydroxypyrrolidine (0.74 M solution in CH2Cl2, 300 mg, 1.7 ml, 1.27 mmol) in CH2C12 (10 ml) and 1 N NaOH (13.0 ml) was added a solution of 3-benzyloxybenzoic acid chloride in anhydrous CH2C12 (5 ml). The resulting mixture was vigorously stirred at room temperature for 3h. The reaction mixture was diluted with EtOAc, washed with brine, and dried over Na2SO4.
The crude product after concentration was triturated in Et2O
and EtOAc to afford white solids (quantitative yield).
To a solution of benzophenone acid (265 mg, 0.39 mmol) in CH2Cl2 (5 ml) was added cat. DMF and oxalyl chloride (2.0 M
solution in CH2C12, 0.488 ml, 0.976 mmol) at room temperature The mixture was kept for stirring at room temperature for 2 hr.
Solvents were removed and the acid chloride residue was taken into CH2Cl2 (5 ml) after drying over the vacuum for lhr.
A solution of amidoalcohol (215.5 mg, 0.39 mmol), Et3N
(197.3 mg, 272 ~L, 1.95 mmol) and DMAP (47.6 mg, 0.39 mmol) in CH2Cl2 (5 ml) was treated with the freshly made acid chloride-CH2Cl2 solution (5 ml) at 5C. The reaction mixture was allowed to stir at room temperature for 3h and then chromatographed on silica gel eluting with 2:3 to l:l/EtOAc:Hexane. The product was obtained as fluffy white solids (332 mg, 70%).
~O 94/20062 215 ~ PCT/US94/02283 The product of the previous reaction (320 mg, 0.264 mmol) was dissolved in EtOAc-HOEt (1:1, 25 ml) and treated with TFA (cat.) followed by 10% Pd(OH)2 (170 mg, 60 mol%). The mixture was subject to hydrogenolysis at 50 psi for 20hr.
Solvents were concentrated after filtering through a pad of celite, and the residue was dissolved in DMF (1.0 ml) and loaded onto HPLC; conditions: A-0.1% TFA 5%CH3CN/H2O, B-100%
CH3CN, 0-50% B over 60 min, 25 ml/min, 41 x 300 mm C18 column.
Fractions (one/min) 37-40 were combined, partially concentrated, and lyophilized to afford fluffy yellow solids (113, 65%). (COMPOUND 675) m.p. 210-213 (dec)-C. IR (KBr) cm 1 3391, 3246, 1717, 1676, 1636, and 1603. Anal. Calcd. for C26H22N2O1l-2-0H2O-1.0C2HF3O2: C, 48.84; H, 3.95; N, 4.07. Found:
C, 48.75; H, 3.63; N, 4.07. LRFAB (M + 1): 579.
wosl/20062 21~ 7 ~ 12 PCT/US94/02283 (+)-Trans-2-t4-(6-hydroxy-2-(carboxyl)benzoyl)-3,5-dihydroxybenzoyloxy]-1-(2-hydroxybenzamido)cyclopentane ~COMPOUND 681) 2 H2,PdonC HO
3.2NKOH,THF h CIOC~3 ~ BnO
BnO
~0 OBn COCI
CH2CI2, DIPEA, DMAP
BnO
Ho2c~ N)~[3 H2 ~0b~1HlB~3 OH OBn FIGURE AP
~2-Benzyloxybenzoyl)chloride To a solution of 2-benzyloxybenzoic acid (684 mg, 3.00 mmol) in CH2Cl2 (15 ml) were added dimethylformamide (1 drop) then oxalyl chloride (3.0 ml of a 2.0 M solution in CH2C12, 6.00 mmol, 2.0 eq). The solution was stirred at room temperature 1 h, then evaporated and the light yellow semi-solid was placed on the vacuum pump before use.
DEMANDES OU BREVETS VOLUMINEUX
LA PRÉSENTE PARTIE DE Ctl I E DENIANDE OU CE BREVET
COMPREND PLUS D'UN TOME. - -CECI EST LE TOME I DE
NOTE: Pour les tomes additionels, veuillez contacter le Bureau canadien desbrevets JU~ABO APPLICATIONS/PATENTS
THIS SECTION OF THE APPI ICATION/PATENT CONTAINS MORE
THAN ONE VOLUME
THIS IS VOLUME L_ OF ~
NOTE: For additional vclumes please c~ntact the Canadian Patent Office
Claims (34)
1. A compound having the formula:
wherein:
A is CH2, NR1, O, S, or SO2.
B1 is NR , CH2 or O;
B2 is CO, CS or SO2;
Z is phenyl, p-hydroxy phenyl, p-benzyloxy phenyl, p-benzoate phenyl, p-carboxy phenyl,4-(2-hydroxyphenylcarbonyl)-3,5-dihydroxy phenyl, p-amino phenyl, 4-fluoro phenyl, 4-benzyloxy phenyl, p-methyl phenyl, p-benzyloxycarbonyl phenyl, p-nitro phenyl, 5-benzyloxy-2-indole, 5-hydroxy-2-indole, 3,4-dihydroxy phenyl, 2-benzyloxy phenyl, 2-hydroxy phenyl, phenyl, p-NHSO2CH3 phenyl, p-methoxymethyleneoxy phenyl, p-acetoxy phenyl;
D is NR3, O or CH2;
E is phenyl, 2-hydroxy benzene, 3-hydroxy benzene, 3-butyloxy benzene, 3-butyloxy-5-hydroxy benzene, 3-hexanoyloxy-5-hydroxy benzene, 3,5-dioctyloxy benzene, 3-octyloxy-5-hydroxy benzene, 3-methoxy-5-hydroxy benzene, 3,5-bis(acetoxy)benzene, 3-(methoxycarbonyl)oxy-5-hydroxybenzene,3,5-dihydroxy phenyl, 3-ethoxy-5-hydroxy phenyl, 3,5-dibenzyloxy phenyl, 3,5-dimethoxy phenyl, 3-hydroxy-5-benzoate phenyl, phenyl, 3,5-dimethoxymethyleneoxy phenyl, 3-methoxycarbonyloxy phenyl, 3-acetoxy-5-hydroxy phenyl;
F is CO or CH2;
G is phenyl, 2-carboxy-6-hydroxy phenyl, 2-ethoxycarbonyl-6-hydroxy phenyl, 2-hydroxy phenyl, 2-benzyloxycarbonyl phenyl, 2-hydroxy naphthyl, 2,3,5,6,-tetramethyl phenyl, 2,6-dihydroxy phenyl, 2,6-dimethoxy phenyl, 2-carboxy cyclohexane, 2-hydroxy cyclohexane, 2-hydroxy-1-naphthyl, 2,6-dichloro phenyl, 2-methoxy-6-hydroxy phenyl, 2-carboxy-3-pyridine, 3-carboxy-2-pyridine, phenyl, 3,4-dibenzyloxyphenylcarbonyl phenyl, 3,4-dihydroxy phenyl, 2-methoxycarbonyl-6-hydroxy phenyl, 2-butoxycarbonyl-6-hydroxy phenyl, 2-(2-methylpropyloxycarbonyl)-6-hydroxy phenyl, 2-nitrilo-6-hydroxy phenyl, 2-carboxy phenyl, 2-(4-acetoxy-benzyloxycarbonyl)-6-hydroxy phenyl, 2-benzyloxycarbonyl -6-benzyloxy phenyl, 2,6-dibenzyloxy phenyl, 2-benzyloxycarbonyl cyclohexane, 1-benzyloxy-2-naphthyl, 2-methoxy-6-benzyloxy phenyl, 2-benzyloxycarbonyl-3-pyridinyl, 3-benzyloxycarbonyl-2-pyridinyl, 2-benzyloxyphenyl, 2-nitrilo-6-benzyloxy phenyl, 3,4-dibenzyloxy phenyl, 2-benzyloxy-1-naphthyl, 6-benzyloxy-2-tetrazolyl phenyl, 6-hydroxy-2-tetrazolyl phenyl, 2-( 2-methyltetrazolyl)-6-hydroxyphenyl, 2-( 3-methyltetrazolyl)-6-hydroxyphenyl, 2-hydroxy-1-(5,6,7,8-tetrahydro) naphthyl, 3-benzyloxycarbonyl-4-benzyloxy phenyl, 3-carboxy-4-hydroxy phenyl, 2-methoxymethyleneoxy phenyl, 2-ethoxycarbonyl-6-benzyloxy phenyl, 2-benzyloxy carbonyl-1-naphthyl, 2-carboxy-1-naphthyl, 2-benzyloxy-6-methyl phenyl, 2-methyl-6-hydroxy phenyl, 2-acetoxy-6-ethoxycarbonyl phenyl, 2-(cyclohexylmethoxycarbonyl)-6-hydroxy phenyl, 2-carboxy-6-benzyloxy phenyl, 2-methoxycarbonyl-6-benzyloxy phenyl, 2-hexanoyloxy-6-carboxy phenyl;
X is CO;
K is H or lower alkyl;
R1, R2, or R3 are, independently hydrogen, lower alkyl or aryl;
m is 1; and n is 3; or, a pharmaceutically acceptable salt thereof.
wherein:
A is CH2, NR1, O, S, or SO2.
B1 is NR , CH2 or O;
B2 is CO, CS or SO2;
Z is phenyl, p-hydroxy phenyl, p-benzyloxy phenyl, p-benzoate phenyl, p-carboxy phenyl,4-(2-hydroxyphenylcarbonyl)-3,5-dihydroxy phenyl, p-amino phenyl, 4-fluoro phenyl, 4-benzyloxy phenyl, p-methyl phenyl, p-benzyloxycarbonyl phenyl, p-nitro phenyl, 5-benzyloxy-2-indole, 5-hydroxy-2-indole, 3,4-dihydroxy phenyl, 2-benzyloxy phenyl, 2-hydroxy phenyl, phenyl, p-NHSO2CH3 phenyl, p-methoxymethyleneoxy phenyl, p-acetoxy phenyl;
D is NR3, O or CH2;
E is phenyl, 2-hydroxy benzene, 3-hydroxy benzene, 3-butyloxy benzene, 3-butyloxy-5-hydroxy benzene, 3-hexanoyloxy-5-hydroxy benzene, 3,5-dioctyloxy benzene, 3-octyloxy-5-hydroxy benzene, 3-methoxy-5-hydroxy benzene, 3,5-bis(acetoxy)benzene, 3-(methoxycarbonyl)oxy-5-hydroxybenzene,3,5-dihydroxy phenyl, 3-ethoxy-5-hydroxy phenyl, 3,5-dibenzyloxy phenyl, 3,5-dimethoxy phenyl, 3-hydroxy-5-benzoate phenyl, phenyl, 3,5-dimethoxymethyleneoxy phenyl, 3-methoxycarbonyloxy phenyl, 3-acetoxy-5-hydroxy phenyl;
F is CO or CH2;
G is phenyl, 2-carboxy-6-hydroxy phenyl, 2-ethoxycarbonyl-6-hydroxy phenyl, 2-hydroxy phenyl, 2-benzyloxycarbonyl phenyl, 2-hydroxy naphthyl, 2,3,5,6,-tetramethyl phenyl, 2,6-dihydroxy phenyl, 2,6-dimethoxy phenyl, 2-carboxy cyclohexane, 2-hydroxy cyclohexane, 2-hydroxy-1-naphthyl, 2,6-dichloro phenyl, 2-methoxy-6-hydroxy phenyl, 2-carboxy-3-pyridine, 3-carboxy-2-pyridine, phenyl, 3,4-dibenzyloxyphenylcarbonyl phenyl, 3,4-dihydroxy phenyl, 2-methoxycarbonyl-6-hydroxy phenyl, 2-butoxycarbonyl-6-hydroxy phenyl, 2-(2-methylpropyloxycarbonyl)-6-hydroxy phenyl, 2-nitrilo-6-hydroxy phenyl, 2-carboxy phenyl, 2-(4-acetoxy-benzyloxycarbonyl)-6-hydroxy phenyl, 2-benzyloxycarbonyl -6-benzyloxy phenyl, 2,6-dibenzyloxy phenyl, 2-benzyloxycarbonyl cyclohexane, 1-benzyloxy-2-naphthyl, 2-methoxy-6-benzyloxy phenyl, 2-benzyloxycarbonyl-3-pyridinyl, 3-benzyloxycarbonyl-2-pyridinyl, 2-benzyloxyphenyl, 2-nitrilo-6-benzyloxy phenyl, 3,4-dibenzyloxy phenyl, 2-benzyloxy-1-naphthyl, 6-benzyloxy-2-tetrazolyl phenyl, 6-hydroxy-2-tetrazolyl phenyl, 2-( 2-methyltetrazolyl)-6-hydroxyphenyl, 2-( 3-methyltetrazolyl)-6-hydroxyphenyl, 2-hydroxy-1-(5,6,7,8-tetrahydro) naphthyl, 3-benzyloxycarbonyl-4-benzyloxy phenyl, 3-carboxy-4-hydroxy phenyl, 2-methoxymethyleneoxy phenyl, 2-ethoxycarbonyl-6-benzyloxy phenyl, 2-benzyloxy carbonyl-1-naphthyl, 2-carboxy-1-naphthyl, 2-benzyloxy-6-methyl phenyl, 2-methyl-6-hydroxy phenyl, 2-acetoxy-6-ethoxycarbonyl phenyl, 2-(cyclohexylmethoxycarbonyl)-6-hydroxy phenyl, 2-carboxy-6-benzyloxy phenyl, 2-methoxycarbonyl-6-benzyloxy phenyl, 2-hexanoyloxy-6-carboxy phenyl;
X is CO;
K is H or lower alkyl;
R1, R2, or R3 are, independently hydrogen, lower alkyl or aryl;
m is 1; and n is 3; or, a pharmaceutically acceptable salt thereof.
2. A compound according to Claim 1 wherein:
A is NH or CH2;
B1 is NH;
B2 is CO;
Z is p-hydroxyphenyl;
D is O;
E is 3,5-dihydroxy benzene;
F is CO;
X is CO;
m is 1; and n is 3.
A is NH or CH2;
B1 is NH;
B2 is CO;
Z is p-hydroxyphenyl;
D is O;
E is 3,5-dihydroxy benzene;
F is CO;
X is CO;
m is 1; and n is 3.
3. A compound having the formula:
wherein:
A is: CH2, NR1, S, SO2, or O;
B1 is : NR, O or CH2;
B2 is: CO, CS, or SO2;
Z is : R4, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
D is: NR3, O or CH2;
E is: R5, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
F is: CO, CS, CH(OR6), CH2, O, S or NR6;
G is: R7, aryl, heteroaryl, substituted aryl, substituted cycloalkyl, or substituted heteroaryl;
K is H or lower alkyl;
X is: CO, CS, CH2, CNR8 or CCR9R10;
R1, R2, R3, R4, R6, R7, R8, R9 and R10 are, independently, hydrogen, lower alkyl, aryl or JR11;
R5 is: lower alkyl or aryl;
J is: CO, C=NH, C=N-lower alkyl, or SO2;
R11 is: lower alkyl, aryl, alkamino, arylamino, aryloxy, or alkoxy;
m is: 1-4;
n is: 1-4; and m plus n is up to 5;
providing that if m is 3, A is NH, B1 is O, B2 is CO, Z is p-hydroxyphenyl, D is NH, X is CO, and E, F, and G, taken together, are then n is not 1, or a pharmaceutically acceptable salt thereof.
wherein:
A is: CH2, NR1, S, SO2, or O;
B1 is : NR, O or CH2;
B2 is: CO, CS, or SO2;
Z is : R4, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
D is: NR3, O or CH2;
E is: R5, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
F is: CO, CS, CH(OR6), CH2, O, S or NR6;
G is: R7, aryl, heteroaryl, substituted aryl, substituted cycloalkyl, or substituted heteroaryl;
K is H or lower alkyl;
X is: CO, CS, CH2, CNR8 or CCR9R10;
R1, R2, R3, R4, R6, R7, R8, R9 and R10 are, independently, hydrogen, lower alkyl, aryl or JR11;
R5 is: lower alkyl or aryl;
J is: CO, C=NH, C=N-lower alkyl, or SO2;
R11 is: lower alkyl, aryl, alkamino, arylamino, aryloxy, or alkoxy;
m is: 1-4;
n is: 1-4; and m plus n is up to 5;
providing that if m is 3, A is NH, B1 is O, B2 is CO, Z is p-hydroxyphenyl, D is NH, X is CO, and E, F, and G, taken together, are then n is not 1, or a pharmaceutically acceptable salt thereof.
4. A compound according to Claim 3 wherein B1 is NH.
5. A compound according to Claim 3 wherein B2 is CO.
6. A compound according to Claim 3 wherein Z is: hydroxy substituted aryl, ether substituted aryl, halo substituted aryl, hydroxy substituted heteroaryl, halo substituted hereroaryl, or ether substituted heteroaryl.
7. A compound according to Claim 3 wherein Z is p-hydroxyphenyl, p-fluorophenyl, or 5-hydroxy indole.
8. A compound according to Claim 3 wherein D is O.
9. A compound according to Claim 3 wherein E is: hydroxy substituted aryl, ether substituted aryl, acyloxy substituted aryl, or hydroxy substituted heteroaryl.
10. A compound according to Claim 3 wherein E is 3,5-dihydroxyphenyl, 3,5-di-lower alkoxyphenyl, 3-hydroxyphenyl, 3,5-diacyloxy phenyl, or 3-acyloxy-5-hydroxyphenyl.
11. A compound according to Claim 3 wherein F is CO.
12. A compound according to Claim 3 wherein G is: hydroxy substituted aryl, ether substituted aryl, carboxyl substituted aryl, tetrazole substituted aryl, cyano substituted aryl, alkoxycarbonyl substituted aryl, acyloxy substituted aryl, hydroxy substituted heteroaryl, ether substituted heteroaryl, carboxyl substituted heteroaryl or combinations of such substitutions.
13. A compound according to Claim 3 wherein G is phenyl, 2-hydroxy phenyl, 2,6-dihydroxy phenyl, 2-lower alkoxy phenyl, 2,6-di-lower alkoxy phenyl, benzene 2-carboxylic acid, 6-hydroxybenzene- 2-carboxylic acid, 2-acyloxy benzene-2-carboxylic acid, 2-hydroxy-6-(2-tetrazoyl)-benzene, 2-hydroxy-6-lower alkoxycarbonyl benzene, 2-acyloxy-6-lower alkoxy carbonyl benzene, 2-hydroxy-6-(trifluoromethylsulfonylamino)-benzene, 2-cyano-6-hydroxybenzene, and 2-hydroxy-5,6,7,8-tetrahydronaphth-1-yl.
14. A compound according to Claim 3 wherein X is CO.
15. A compound according to Claim 3 where m is 1 or 2.
16. A compound according to Claim 3 where n is 1, 2 or 3.
17. A compound according to Claim 3 where n is 1 and m is 1.
18. A compound according to Claim 3 where n is 1 and m is 3.
19. A method of inhibiting protein kinase C comprising contacting protein kinase C with an effective amount of a compound in accordance with claim 1 or 3.
20. A method of inhibiting protein kinase C comprising contacting protein kinase C with an effective amount of a compound having the formula:
wherein:
A is CH2, NR1, S, SO2, or O;
B1 is NR2, O or CH2;
B2 is CO, CS, or SO2;
Z is R4, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
D is NR3, O or CH2;
E is R5, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
F is CO, CS, CH(OR6), CH2, O, S or NR6;
G is R7, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
K is H or lower alkyl;
X is CO, CS, CH2, CNR8 or CCR9R10;
R1 is hydrogen, lower alkyl, aryl or JR11;
J is CO, C=NH or SO2;
R11 is lower alkyl, aryl, alkylamino or alkoxy;
R2, R3, R4, R6, R7, R8, R9 and R10 are, independently hydrogen, lower alkyl or aryl;
R5 is lower alkyl or aryl;
R11 is lower alkyl, aryl, alkamino, arylamino, aryloxy, or alkoxy;
m is 1-4; and n is 1-4;
wherein m plus n is less than or equal to 5; or, a pharmaceutically acceptable salt thereof.
wherein:
A is CH2, NR1, S, SO2, or O;
B1 is NR2, O or CH2;
B2 is CO, CS, or SO2;
Z is R4, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
D is NR3, O or CH2;
E is R5, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
F is CO, CS, CH(OR6), CH2, O, S or NR6;
G is R7, aryl, heteroaryl, substituted aryl or substituted heteroaryl;
K is H or lower alkyl;
X is CO, CS, CH2, CNR8 or CCR9R10;
R1 is hydrogen, lower alkyl, aryl or JR11;
J is CO, C=NH or SO2;
R11 is lower alkyl, aryl, alkylamino or alkoxy;
R2, R3, R4, R6, R7, R8, R9 and R10 are, independently hydrogen, lower alkyl or aryl;
R5 is lower alkyl or aryl;
R11 is lower alkyl, aryl, alkamino, arylamino, aryloxy, or alkoxy;
m is 1-4; and n is 1-4;
wherein m plus n is less than or equal to 5; or, a pharmaceutically acceptable salt thereof.
21. The method according to Claim 20 wherein:
A is NH, CH2 or NR1.
B1 is NR2 or O;
B2 is CO or CS;
Z is phenyl or hydroxy benzene;
D is NR3, O or CH2;
E is 3,5-hydroxy benzene or 3,5-methoxy benzene;
F is CO or CH2;
G is phenyl, 2-hydroxy phenyl, 2,6-dihydroxy phenyl, 2-lower alkoxy phenyl, 2,6-di-lower alkoxy phenyl, benzene 2-carboxylic acid, 6-hydroxybenzene- 2-carboxylic acid, 2-acyloxy benzene-2-carboxylic acid, 2-hydroxy-6-(2-tetrazoyl)-benzene, 2-hydroxy-6-lower alkoxycarbonyl benzene, 2-acyloxy-6-lower alkoxy carbonyl benzene, 2-hydroxy-6-(trifluoromethylsulfonyl amino)-benzene, 2-cyano-6-hydroxybenzene, and 2-hydroxy-5,6,7,8-tetrahydronaphth-1-yl.
X is CO;
R1, R2, or R3 are, independently hydrogen, lower alkyl or aryl;
m is 1; and n is 3;
or, a pharmaceutically acceptable salt thereof.
A is NH, CH2 or NR1.
B1 is NR2 or O;
B2 is CO or CS;
Z is phenyl or hydroxy benzene;
D is NR3, O or CH2;
E is 3,5-hydroxy benzene or 3,5-methoxy benzene;
F is CO or CH2;
G is phenyl, 2-hydroxy phenyl, 2,6-dihydroxy phenyl, 2-lower alkoxy phenyl, 2,6-di-lower alkoxy phenyl, benzene 2-carboxylic acid, 6-hydroxybenzene- 2-carboxylic acid, 2-acyloxy benzene-2-carboxylic acid, 2-hydroxy-6-(2-tetrazoyl)-benzene, 2-hydroxy-6-lower alkoxycarbonyl benzene, 2-acyloxy-6-lower alkoxy carbonyl benzene, 2-hydroxy-6-(trifluoromethylsulfonyl amino)-benzene, 2-cyano-6-hydroxybenzene, and 2-hydroxy-5,6,7,8-tetrahydronaphth-1-yl.
X is CO;
R1, R2, or R3 are, independently hydrogen, lower alkyl or aryl;
m is 1; and n is 3;
or, a pharmaceutically acceptable salt thereof.
22. The method according to Claim 20 wherein:
A is NH or CH2;
B1 is NH;
B2 is CO;
Z is p-hydroxyphenyl;
D is O;
E is 3,5-hydroxy benzene;
F is CO;
G is phenyl, 2-hydroxy phenyl, 2,6-dihydroxy phenyl, 2-lower alkoxy phenyl, 2,6-di-lower alkoxy phenyl, benzene 2-carboxylic acid, 6-hydroxybenzene- 2-carboxylic acid, 2-acyloxy benzene-2-carboxylic acid, 2-hydroxy-6-(2-tetrazoyl)-benzene, 2-hydroxy-6-lower alkoxycarbonyl benzene, 2-acyloxy-6-lower alkoxy carbonyl benzene, 2-hydroxy-6-(trifluoromethylsulfonyl amino)-benzene, 2-cyano-6-hydroxybenzene, and 2-hydroxy-5,6,7,8-tetrahydronaphth-1-yl.
X is CO;
m is 1; and n is 3.
A is NH or CH2;
B1 is NH;
B2 is CO;
Z is p-hydroxyphenyl;
D is O;
E is 3,5-hydroxy benzene;
F is CO;
G is phenyl, 2-hydroxy phenyl, 2,6-dihydroxy phenyl, 2-lower alkoxy phenyl, 2,6-di-lower alkoxy phenyl, benzene 2-carboxylic acid, 6-hydroxybenzene- 2-carboxylic acid, 2-acyloxy benzene-2-carboxylic acid, 2-hydroxy-6-(2-tetrazoyl)-benzene, 2-hydroxy-6-lower alkoxycarbonyl benzene, 2-acyloxy-6-lower alkoxy carbonyl benzene, 2-hydroxy-6-(trifluoromethylsulfonyl amino)-benzene, 2-cyano-6-hydroxybenzene, and 2-hydroxy-5,6,7,8-tetrahydronaphth-1-yl.
X is CO;
m is 1; and n is 3.
23. A compound having the formula:
Wherein R12 is 4-hydroxyphenyl, 4-lower alkoxyphenyl, 4-fluorophenyl, 5-hydroxy-2-indolyl, and 4-acetoxyphenyl; R13 is 2-carboxy-6-hydroxyphenyl, 2,6-dihydroxyphenyl, 2,6-di-lower alkoxyphenyl, 2-carboxy-6-lower alkoxyphenyl, 2-hydroxy-5,6,7,8-tetrahydronaphthyl, 2-methoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-hydroxy phenyl, 2-lower alkoxycarbonyl-6-hydroxyphenyl, 2-butoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-acetoxyphenyl, 2-hydroxy-1-naphthyl, COCH2OCOC(CH3)3, 2-tetrazolyl-6-hydroxyphenyl; and R14 is H, methylsulfonyl, lower alkylsulfonyl, beta naphthylsulfonyl, 4-toluene sulfonyl, tert-butoxy carbonyl, lower alkoxy carbonyl, (CH3CH2O)2PO-, methyl, ethyl, propyl, isopropyl, lower alkyl, tert-butylimino; K is H or lower alkyl; and salts thereof.
Wherein R12 is 4-hydroxyphenyl, 4-lower alkoxyphenyl, 4-fluorophenyl, 5-hydroxy-2-indolyl, and 4-acetoxyphenyl; R13 is 2-carboxy-6-hydroxyphenyl, 2,6-dihydroxyphenyl, 2,6-di-lower alkoxyphenyl, 2-carboxy-6-lower alkoxyphenyl, 2-hydroxy-5,6,7,8-tetrahydronaphthyl, 2-methoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-hydroxy phenyl, 2-lower alkoxycarbonyl-6-hydroxyphenyl, 2-butoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-acetoxyphenyl, 2-hydroxy-1-naphthyl, COCH2OCOC(CH3)3, 2-tetrazolyl-6-hydroxyphenyl; and R14 is H, methylsulfonyl, lower alkylsulfonyl, beta naphthylsulfonyl, 4-toluene sulfonyl, tert-butoxy carbonyl, lower alkoxy carbonyl, (CH3CH2O)2PO-, methyl, ethyl, propyl, isopropyl, lower alkyl, tert-butylimino; K is H or lower alkyl; and salts thereof.
24. A compound having the formula:
wherein R15 is 4-hydroxyphenyl, 4-lower alkoxyphenyl, 4-fluorophenyl, 5-hydroxy-2-indolyl, and 4-acetoxyphenyl; R16 is 2-carboxy-6-hydroxyphenyl, 2,6-dihydroxyphenyl, 2,6-di-lower alkoxyphenyl, 2-carboxy-6-lower alkoxyphenyl, 2-hydroxy-5,6,7,8-tetrahydronaphthyl,2-methoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-hydroxy phenyl, 2-lower alkoxycarbonyl-6-hydroxyphenyl, 2-butoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-acetoxyphenyl, 2-hydroxy-1-naphthyl, COCH2OCOC(CH3)3, 2-tetrazolyl-6-hydroxyphenyl; and R17 is H, methylsulfonyl, lower alkylsulfonyl, beta naphthylsulfonyl, 4-toluene sulfonyl, tert-butoxy carbonyl, lower alkoxy carbonyl, (CH3CH2O)2PO-, methyl, ethyl, propyl, isopropyl, lower alkyl, tert-butylimino, K is H or lower alkyl; and salts thereof.
wherein R15 is 4-hydroxyphenyl, 4-lower alkoxyphenyl, 4-fluorophenyl, 5-hydroxy-2-indolyl, and 4-acetoxyphenyl; R16 is 2-carboxy-6-hydroxyphenyl, 2,6-dihydroxyphenyl, 2,6-di-lower alkoxyphenyl, 2-carboxy-6-lower alkoxyphenyl, 2-hydroxy-5,6,7,8-tetrahydronaphthyl,2-methoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-hydroxy phenyl, 2-lower alkoxycarbonyl-6-hydroxyphenyl, 2-butoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-acetoxyphenyl, 2-hydroxy-1-naphthyl, COCH2OCOC(CH3)3, 2-tetrazolyl-6-hydroxyphenyl; and R17 is H, methylsulfonyl, lower alkylsulfonyl, beta naphthylsulfonyl, 4-toluene sulfonyl, tert-butoxy carbonyl, lower alkoxy carbonyl, (CH3CH2O)2PO-, methyl, ethyl, propyl, isopropyl, lower alkyl, tert-butylimino, K is H or lower alkyl; and salts thereof.
25. A compound having the formula:
wherein R18 is 4-hydroxyphenyl, 4-lower alkoxyphenyl, 4-fluorophenyl, 5-hydroxy-2-indolyl, and 4-acetoxyphenyl; and R19 is 2-carboxy-6-hydroxyphenyl, 2,6-dihydroxyphenyl, 2,6-dicarboxyphenyl, 2,6-di-lower alkoxyphenyl, 2-carboxy-6-lower alkoxyphenyl, 2-hydroxy-5,6,7,8-tetrahydronaphthyl, 2-methoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-hydroxyphenyl, 2-lower alkoxycarbonyl-6-hydroxyphenyl, 2,6-dilower alkoxycarbonylphenyl, 2-butoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-acetoxyphenyl, 2-hydroxy-1-naphthyl, COCH2CH2OCOC(CH3)3, 2-tetrazolyl-6-hydroxyphenyl; K is H or lower alkyl; and salts thereof.
wherein R18 is 4-hydroxyphenyl, 4-lower alkoxyphenyl, 4-fluorophenyl, 5-hydroxy-2-indolyl, and 4-acetoxyphenyl; and R19 is 2-carboxy-6-hydroxyphenyl, 2,6-dihydroxyphenyl, 2,6-dicarboxyphenyl, 2,6-di-lower alkoxyphenyl, 2-carboxy-6-lower alkoxyphenyl, 2-hydroxy-5,6,7,8-tetrahydronaphthyl, 2-methoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-hydroxyphenyl, 2-lower alkoxycarbonyl-6-hydroxyphenyl, 2,6-dilower alkoxycarbonylphenyl, 2-butoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-acetoxyphenyl, 2-hydroxy-1-naphthyl, COCH2CH2OCOC(CH3)3, 2-tetrazolyl-6-hydroxyphenyl; K is H or lower alkyl; and salts thereof.
26. A compound having the formula:
Wherein R12 is 4-hydroxyphenyl, 4-fluorophenyl, 5-hydroxy-2-indolyl: R13 is 2-carboxy-6-hydroxyphenyl, 2,6-dihydroxyphenyl;
and R14 is H, methylsulfonyl, beta naphthylsulfonyl, 4-toluene sulfonyl, tert-butoxy carbonyl; and salts thereof.
Wherein R12 is 4-hydroxyphenyl, 4-fluorophenyl, 5-hydroxy-2-indolyl: R13 is 2-carboxy-6-hydroxyphenyl, 2,6-dihydroxyphenyl;
and R14 is H, methylsulfonyl, beta naphthylsulfonyl, 4-toluene sulfonyl, tert-butoxy carbonyl; and salts thereof.
27. A compound having the formula:
wherein R15 is 4-hydroxyphenyl, 4-acetoxyphenyl; R16 is 2-carboxy-6-hydroxyphenyl,2-hydroxy-5,6,7,8-tetrahydronaphthyl, 2-methoxycarbonyl-6-hydroxyphenyl, 2-butoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-acetoxyphenyl, 2-hydroxy-1-naphthyl; and R17 is H, (CH3CH2O)2PO-, isopropyl, tert-butylidinyl, and salts thereof.
wherein R15 is 4-hydroxyphenyl, 4-acetoxyphenyl; R16 is 2-carboxy-6-hydroxyphenyl,2-hydroxy-5,6,7,8-tetrahydronaphthyl, 2-methoxycarbonyl-6-hydroxyphenyl, 2-butoxycarbonyl-6-hydroxyphenyl, 2-ethoxycarbonyl-6-acetoxyphenyl, 2-hydroxy-1-naphthyl; and R17 is H, (CH3CH2O)2PO-, isopropyl, tert-butylidinyl, and salts thereof.
28. A compound having the formula:
wherein R18 is 4-hydroxyphenyl; and R19 is COCH2OCOC(CH3)3, 2-tetrazolyl-6-hydroxyphenyl, 2-carboxy-6-hydroxyphenyl, 2-methoxycarbonyl-6-hydroxyphenyl; and salts thereof.
wherein R18 is 4-hydroxyphenyl; and R19 is COCH2OCOC(CH3)3, 2-tetrazolyl-6-hydroxyphenyl, 2-carboxy-6-hydroxyphenyl, 2-methoxycarbonyl-6-hydroxyphenyl; and salts thereof.
29. A compound in accordance with claim 24, 25, 26, 27, 28, or 29 in a pharmaceutically acceptable carrier or diluent.
30. A method of inhibiting protein kinase C comprising contacting protein kinase C with an effective amount of a compound in accordance with claim 24, 25, 26, 27, 28, or 29.
31. A method of treating an inflammatory disease comprising administering to a mammal suspected of having an inflammatory disease an effective amount of a compound in accordance with claim 24, 25, 26, 27, 28, or 29.
32. (-)-Trans-4-(4-(2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxy benzoyloxy)-3-(4-hydroxybenzamido)azepine Trifluoroacetic Acid Salt, (-)-Balanol; syn-4-[4-(2-hydroxycarbonyl-6-hydroxy benzoyl)-3,5-dihydroxy benzoyloxy]-3-(4-hydroxybenzamido) perhydroazepine trifluoroacetic acid salt;
trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-hydroxycarbonyl benzoyl)-3,5-dihydroxybenzoyloxyl]pyrrolidinium trifluoro acetate; trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-naphth-1-oyl)-3,5-dihydroxybenzyoyloxyl]pyrrolidine trifluoroacetic acid salt;(+)-Anti-4-[3,5-dihydroxy-4-(2,6-dihydroxy benzoyl)]
hexahydro-3-(4-hydroxybenzoylamine)azepine; 1-Isopropyl-trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-hydroxycarbonyl benzoyl)-3,5-dihydroxybenzoyloxyl]pyrrolidinium trifluoro acetate; anti-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-1-[5,6,7,8-tetrahydro-naphthoyl])-3,5-dihydroxy benzyoyl oxy]pyrrolidine trifluoroacetic acid salt; anti-1-[4-(2-Hydroxycarbonyl-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy]-2-(4-hydroxybenzamido) cyclopentane; ()-trans-3-(4-hydroxy benzamido)-4-[4-(2-hydroxy-6-methoxycarbonyl benzoyl)-3,5-dihydroxybenzoyloxy]pyrrolidinium hydrochloride; racemic tosyl-balanol;trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-hydroxycarbonylbenzoyl)-3,5-dihydroxybenzamido] pyrrolidine trifluoroacetic acid salt; ()-trans-3-(4-acetoxy benzamido)-4[4-(2-acetoxy-6-ethoxycarbonylbenzoyl)-3-acetoxy-5-hydroxy benzoyloxy] pyrrolidinium trifluoroacetate; ()anti-1-[4-(2-Methoxycarbonyl-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy]-2-(4-hydroxybenzamido)-cyclopentane; ()-trans-4-[4-(2-hydroxy carbonyl-6-hydroxybenzoyl)-3,5-dihydroxy benzoyloxy]-3-(4-hydroxybenzamido)-1-(2-naphthalene sulfonyl) azepine; ()-1-1-methylethyl-trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-methoxycarbonylbenzoyl)-3,5-dihydroxybenzoyl oxy]pyrrolidine trifluoroacetic acid salt; trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-butoxycarbonylbenzoyl)-3,5-dihyrdroxybenzoyloxy]
pyrrolidine trifluoroacetic acid salt; (+)-trans-3-(4-Hydroxy benzamido)-4-[3,5-dihydroxy-4-(2- carboxy-6-hydroxybenzoyl benzoyloxy]-N-diethylphosphonato pyrrolidine; ()-trans-2-[4-(6-hydroxy-2-tetrazolylbenzoyl)-3,5-dihydroxybenzoyloxy]-1-(4-hydroxybenzamido)cyclopentane; ()-trans-3-(4-hydroxybenzamido) -4-[4-(2-hydroxy-6-methoxycarbonylbenzoyl)-3,5-dihydroxy benzamido] pyrrolidine trifluoroacetic acid salt; mesyl-balanol; 1-[4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxy benzyloxy]-2-(4-hydroxybenzamido) cyclopentane; (+) -trans-3-(4-Fluorobenzamido)-4-[4-(2-carboxy-6-hydroxy) benzoyl-3,5-dihydroxy]benzoyloxyhexahydroazepine trifluoroacetic acid salt; ()-anti-3-[4-(2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxy benzoyl-oxyl-1-[N(1,1-dimethyl ethyl)iminomethyl]-4-(4-hydroxybenzamido)-pyrrolidine, trifluoroacetic acid salt; BOC-Balanol; (+)-trans-4-[4-(2-carboxy-6-hydroxy)benzoyl-3,5-dihydroxy]benzoyloxy-3-[2-(5-hydroxyindolyl)formamido]-hexahydroazepine trifluoroacetic acid salt; or ()-trans-2-[3,5-Dihydroxy-4-(2-hydroxy-6-(trifluoro methane-sulfonylamino) benzoyl)benzoyloxy]-1-(4-hydroxy benzamido)-cyclopentane Hemihydrate; or a salt or pharmaceutically acceptable solution, suspension, or dispersion thereof.
trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-hydroxycarbonyl benzoyl)-3,5-dihydroxybenzoyloxyl]pyrrolidinium trifluoro acetate; trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-naphth-1-oyl)-3,5-dihydroxybenzyoyloxyl]pyrrolidine trifluoroacetic acid salt;(+)-Anti-4-[3,5-dihydroxy-4-(2,6-dihydroxy benzoyl)]
hexahydro-3-(4-hydroxybenzoylamine)azepine; 1-Isopropyl-trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-hydroxycarbonyl benzoyl)-3,5-dihydroxybenzoyloxyl]pyrrolidinium trifluoro acetate; anti-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-1-[5,6,7,8-tetrahydro-naphthoyl])-3,5-dihydroxy benzyoyl oxy]pyrrolidine trifluoroacetic acid salt; anti-1-[4-(2-Hydroxycarbonyl-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy]-2-(4-hydroxybenzamido) cyclopentane; ()-trans-3-(4-hydroxy benzamido)-4-[4-(2-hydroxy-6-methoxycarbonyl benzoyl)-3,5-dihydroxybenzoyloxy]pyrrolidinium hydrochloride; racemic tosyl-balanol;trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-hydroxycarbonylbenzoyl)-3,5-dihydroxybenzamido] pyrrolidine trifluoroacetic acid salt; ()-trans-3-(4-acetoxy benzamido)-4[4-(2-acetoxy-6-ethoxycarbonylbenzoyl)-3-acetoxy-5-hydroxy benzoyloxy] pyrrolidinium trifluoroacetate; ()anti-1-[4-(2-Methoxycarbonyl-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy]-2-(4-hydroxybenzamido)-cyclopentane; ()-trans-4-[4-(2-hydroxy carbonyl-6-hydroxybenzoyl)-3,5-dihydroxy benzoyloxy]-3-(4-hydroxybenzamido)-1-(2-naphthalene sulfonyl) azepine; ()-1-1-methylethyl-trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-methoxycarbonylbenzoyl)-3,5-dihydroxybenzoyl oxy]pyrrolidine trifluoroacetic acid salt; trans-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-butoxycarbonylbenzoyl)-3,5-dihyrdroxybenzoyloxy]
pyrrolidine trifluoroacetic acid salt; (+)-trans-3-(4-Hydroxy benzamido)-4-[3,5-dihydroxy-4-(2- carboxy-6-hydroxybenzoyl benzoyloxy]-N-diethylphosphonato pyrrolidine; ()-trans-2-[4-(6-hydroxy-2-tetrazolylbenzoyl)-3,5-dihydroxybenzoyloxy]-1-(4-hydroxybenzamido)cyclopentane; ()-trans-3-(4-hydroxybenzamido) -4-[4-(2-hydroxy-6-methoxycarbonylbenzoyl)-3,5-dihydroxy benzamido] pyrrolidine trifluoroacetic acid salt; mesyl-balanol; 1-[4-(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxy benzyloxy]-2-(4-hydroxybenzamido) cyclopentane; (+) -trans-3-(4-Fluorobenzamido)-4-[4-(2-carboxy-6-hydroxy) benzoyl-3,5-dihydroxy]benzoyloxyhexahydroazepine trifluoroacetic acid salt; ()-anti-3-[4-(2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxy benzoyl-oxyl-1-[N(1,1-dimethyl ethyl)iminomethyl]-4-(4-hydroxybenzamido)-pyrrolidine, trifluoroacetic acid salt; BOC-Balanol; (+)-trans-4-[4-(2-carboxy-6-hydroxy)benzoyl-3,5-dihydroxy]benzoyloxy-3-[2-(5-hydroxyindolyl)formamido]-hexahydroazepine trifluoroacetic acid salt; or ()-trans-2-[3,5-Dihydroxy-4-(2-hydroxy-6-(trifluoro methane-sulfonylamino) benzoyl)benzoyloxy]-1-(4-hydroxy benzamido)-cyclopentane Hemihydrate; or a salt or pharmaceutically acceptable solution, suspension, or dispersion thereof.
33. 3-Benzyloxy-2-[2,6-dibenzyloxy-4-(1,1-dimethylethoxy carbonyl)benzoyl]benzoic acid or a lower alkyl ester thereof.
34. 3,5-dimethoxymethyleneoxy-4-[2-methoxymethyleneoxy-6-(1,6-dioxanyl)]benzoylbenzoic acid or a lower alkyl ester thereof.
Applications Claiming Priority (2)
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| US2584693A | 1993-03-03 | 1993-03-03 | |
| US08/025,846 | 1993-03-03 |
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| AU (1) | AU6252794A (en) |
| CA (1) | CA2157412A1 (en) |
| IL (1) | IL108849A0 (en) |
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| TW339325B (en) * | 1995-07-05 | 1998-09-01 | Hoffmann La Roche | Novel azepane derivatives, process for the preparation thereof and pharmaceutical composition containing the same |
| US6914140B1 (en) | 1996-04-17 | 2005-07-05 | Hoffmann-La Roche Inc. | Asymmetric synthesis process |
| EP0802190A1 (en) * | 1996-04-17 | 1997-10-22 | F. Hoffmann-La Roche Ag | Process and intermediates for preparing azepines |
| US5902882A (en) * | 1996-04-17 | 1999-05-11 | Hoffmann-La Roche Inc. | Assymetric synthesis of azepines |
| CN1261098C (en) | 1998-08-28 | 2006-06-28 | 西奥斯股份有限公司 | Use of piperidines and/or piperazines as inhibitors of p38-alpha kinase |
| DZ3083A1 (en) * | 1999-02-19 | 2004-06-02 | Smithkline Beecham Corp | New protease inhibitor compounds, process for their preparation and pharmaceutical compositions containing them |
| US6646005B1 (en) | 1999-06-01 | 2003-11-11 | The University Of Texas Southwestern Medical Center | Method of treating hair loss using sulfonyl thyromimetic compounds |
| US6680344B1 (en) | 1999-06-01 | 2004-01-20 | The University Of Texas Southwestern Medical Center | Method of treating hair loss using diphenylmethane derivatives |
| MXPA01012493A (en) | 1999-06-01 | 2003-09-10 | Univ Texas Southwestern Med Ct | Method of treating hair loss using diphenylether derivatives. |
| US6541477B2 (en) | 1999-08-27 | 2003-04-01 | Scios, Inc. | Inhibitors of p38-a kinase |
| US6608227B1 (en) | 1999-10-15 | 2003-08-19 | Bristol-Myers Squibb Pharma | Benzylcycloalkyl amines as modulators of chemokine receptor activity |
| TWI288745B (en) | 2000-04-05 | 2007-10-21 | Daiichi Seiyaku Co | Ethylenediamine derivatives |
| HUP0303652A2 (en) | 2000-12-20 | 2004-03-01 | Bristol-Myers Squibb Company | Cyclic derivatives as modulators of chemokine receptor activity and pharmaceutical compositions containing them |
| CZ20032258A3 (en) | 2001-02-23 | 2004-01-14 | Merck & Co., Inc. | N-substituted non-aryl heterocyclic compounds |
| JP2005511478A (en) | 2001-04-03 | 2005-04-28 | メルク エンド カムパニー インコーポレーテッド | N-substituted non-aryl heterocyclic amidyl NMDA / NR2B antagonist |
| WO2003000657A1 (en) | 2001-06-20 | 2003-01-03 | Daiichi Pharmaceutical Co., Ltd. | Diamine derivatives |
| CN1826333B (en) * | 2001-06-20 | 2012-12-26 | 第一三共株式会社 | Diamine derivatives |
| JP4331595B2 (en) * | 2001-08-09 | 2009-09-16 | 第一三共株式会社 | Diamine derivatives |
| KR100652451B1 (en) | 2001-09-13 | 2006-12-01 | 에프. 호프만-라 로슈 아게 | Ccr-3 receptor antagonists v |
| AU2003218028A1 (en) | 2002-03-08 | 2003-09-22 | Bristol-Myers Squibb Company | Cyclic derivatives as modulators of chemokine receptor activity |
| US6887864B2 (en) | 2002-03-12 | 2005-05-03 | Hoffmann-La Roche Inc. | Azepane derivatives |
| JP2017518312A (en) * | 2014-06-05 | 2017-07-06 | ユニバーシティ・オブ・カンザス | Malmelin analogs and methods of use in cancer treatment |
| WO2019089729A1 (en) | 2017-10-31 | 2019-05-09 | University Of Miami | Kinase inhibitors for the treatment of central and peripheral nervous system disorders |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2504192A (en) * | 1991-08-22 | 1993-03-16 | Sphinx Pharmaceuticals Corporation | Protein kinase c inhibition and novel compound balanol |
-
1994
- 1994-03-02 WO PCT/US1994/002283 patent/WO1994020062A2/en not_active Application Discontinuation
- 1994-03-02 AU AU62527/94A patent/AU6252794A/en not_active Abandoned
- 1994-03-02 EP EP94909847A patent/EP0687249A1/en not_active Withdrawn
- 1994-03-02 JP JP6520148A patent/JPH09503994A/en active Pending
- 1994-03-02 CA CA002157412A patent/CA2157412A1/en not_active Abandoned
- 1994-03-03 IL IL10884994A patent/IL108849A0/en unknown
- 1994-03-03 ZA ZA941478A patent/ZA941478B/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| WO1994020062A2 (en) | 1994-09-15 |
| WO1994020062A3 (en) | 1996-08-15 |
| JPH09503994A (en) | 1997-04-22 |
| AU6252794A (en) | 1994-09-26 |
| IL108849A0 (en) | 1994-06-24 |
| EP0687249A1 (en) | 1995-12-20 |
| ZA941478B (en) | 1995-09-05 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Discontinued |