AU709239B2 - Aspartyl protease inhibitors - Google Patents
Aspartyl protease inhibitors Download PDFInfo
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- AU709239B2 AU709239B2 AU17580/97A AU1758097A AU709239B2 AU 709239 B2 AU709239 B2 AU 709239B2 AU 17580/97 A AU17580/97 A AU 17580/97A AU 1758097 A AU1758097 A AU 1758097A AU 709239 B2 AU709239 B2 AU 709239B2
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- 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
- C07D207/18—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 one double bond between ring members or between a ring member and a non-ring member
- C07D207/22—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 one double bond between ring members or between a ring member and a non-ring member 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
- C07D207/24—Oxygen or sulfur atoms
- C07D207/26—2-Pyrrolidones
- C07D207/263—2-Pyrrolidones with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms
- C07D207/27—2-Pyrrolidones with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms with substituted hydrocarbon radicals directly attached to the ring nitrogen atom
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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- 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
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
- C07D233/04—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
- C07D233/28—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member 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
- C07D233/30—Oxygen or sulfur atoms
- C07D233/32—One oxygen atom
- C07D233/36—One oxygen atom with hydrocarbon radicals, substituted by nitrogen atoms, attached to ring nitrogen atoms
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- C—CHEMISTRY; METALLURGY
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- C07D265/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one oxygen atom as the only ring hetero atoms
- C07D265/28—1,4-Oxazines; Hydrogenated 1,4-oxazines
- C07D265/30—1,4-Oxazines; Hydrogenated 1,4-oxazines not condensed with other rings
- C07D265/32—1,4-Oxazines; Hydrogenated 1,4-oxazines not condensed with other rings with oxygen atoms directly attached to ring carbon atoms
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- C07D265/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one oxygen atom as the only ring hetero atoms
- C07D265/28—1,4-Oxazines; Hydrogenated 1,4-oxazines
- C07D265/34—1,4-Oxazines; Hydrogenated 1,4-oxazines condensed with carbocyclic rings
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D285/00—Heterocyclic compounds containing rings having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by groups C07D275/00 - C07D283/00
- C07D285/01—Five-membered rings
- C07D285/02—Thiadiazoles; Hydrogenated thiadiazoles
- C07D285/04—Thiadiazoles; Hydrogenated thiadiazoles not condensed with other rings
- C07D285/10—1,2,5-Thiadiazoles; Hydrogenated 1,2,5-thiadiazoles
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- 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
- C07D401/06—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 carbon chain containing only aliphatic carbon atoms
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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
- C07D401/14—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 three or more hetero rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
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- C—CHEMISTRY; METALLURGY
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- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/06—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero 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
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
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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
- C07D417/06—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 carbon chain containing only aliphatic carbon atoms
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- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
- C07D491/10—Spiro-condensed systems
Description
WO 97/27180 PCT/US97/01610 ASPARTYL PROTEASE INHIBITORS TECHNICAL FIELD OF THE INVENTION The present invention relates to a novel class of compounds which are aspartyl protease inhibitors. In one embodiment, this invention relates to a novel class of HIV aspartyl protease inhibitors characterized by specific structural and physicochemical features. This invention also relates to pharmaceutical compositions comprising these compounds. The compounds and pharmaceutical compositions of this invention are particularly well suited for inhibiting HIV-1 and HIV-2 protease activity and consequently, may be advantageously used as antiviral agents against the HIV-1 and HIV-2 viruses. This invention also relates to methods for inhibiting aspartyl protease activity, methods for treating viral infections using the compounds and compositions of this invention, and methods for making intermediates and compounds of this invention.
WO 97/27180 PCT/US97/01610 2 BACKGROUND OF THE INVENTION The human immunodeficiency virus is the causative agent for acquired immunodeficiency syndrome ("AIDS") a disease characterized by the destruction of the immune system, particularly of CD4 T-cells, with attendant susceptibility to opportunistic infections and its precursor AIDS-related complex a syndrome characterized by symptoms such as persistent generalized lymphadenopathy, fever and weight loss.
As in the case of several other retroviruses, HIV encodes the production of a protease which carries out post-translational cleavage of precursor polypeptides in a process necessary for the formation of infectious virions Crawford et al., "A Deletion Mutation in the 5' Part of the pol Gene of Moloney Murine Leukemia Virus Blocks Proteolytic Processing of the gag and pol Polyproteins", J. Virol., 53, p. 899 (1985)). These gene products include pol, which encodes the virion RNA-dependent DNA polymerase (reverse transcriptase), an endonuclease, HIV protease, and gag, which encodes the core-proteins of the virion Toh et al., "Close Structural Resemblance Between Putative Polymerase of a Drosophila Transposable Genetic Element 17.6 and pol gene product of Moloney Murine Leukemia Virus", EMBO 4, p. 1267 (1985); L.H. Pearl et al., "A Structural Model for the Retroviral Proteases", Nature, pp. 329-351 (1987); M.D.
Power et al., "Nucleotide Sequence of SRV-1, a Type D Simian Acquired Immune Deficiency Syndrome Retrovirus", Science, 231, p. 1567 (1986)).
A number of synthetic anti-viral agents have been designed to target various stages in the WO 97/27180 PCT/US97/01610 3 replication cycle of HIV. These agents include compounds which block viral.binding to CD4 Tlymphocytes (for example, soluble CD4), and compounds which interfere with viral replication by inhibiting viral reverse transcriptase (for example, didanosine and zidovudine (AZT)) and inhibit integration of viral DNA into cellular DNA Hirsh and R.T. D'Aqulia, "Therapy for Human Immunodeficiency Virus Infection", N.Ena.J.Med., 328, p. 1686 (1993)). However, such agents, which are directed primarily to early stages of viral replication, do not prevent the production of infectious virions in chronically infected cells.
Furthermore, administration of some of these agents in effective amounts has led to cell-toxicity and unwanted side effects, such as anemia and bone marrow suppression.
More recently, drug design efforts have been directed toward creating compounds which inhibit the formation of infectious virions by interfering with the processing of viral polyprotein precursors. Processing of these precursor proteins requires the action of virus-encoded proteases which are essential for replication (Kohl, N.E. et al. "Active HIV Protease is Required for Viral Infectivity" Proc. Natl. Acad. Sci.
USA, 85, p. 4686 (1988)). The anti-viral potential of HIV protease inhibition has been demonstrated using peptidal inhibitors. Such peptidal compounds, however, are typically large and complex molecules that tend to exhibit poor bioavailability and are not generally consistent with oral administration. Accordingly, the need still exists for compounds that can effectively inhibit the action of viral proteases, for use as agents for preventing and treating chronic and acute viral infections. Such agents would be expected to act PCT/US97/01610 VOSS1US PARTNER Vertex Pharmaceuticals Inc., et al FATENTAN'ANALTE.
Our Ref.: B 2555 PCT 3IEBERTSTR.4 81 6 75 MONCHEN 4 19. Feb. 1998 as effective therapeutic agents in their own right. In addition, since they act at a separate stage in the virus life cycle from previously described antiretroviral agents, the administration of a combination of agents would be expected to result in increased therapeutic efficacy.
International Publication WO 94/19329 discloses cyclic carbonyls and derivatives thereof as protease inhibitors. International Publication WO 95/24385 discloses sulfonamide protease inhibitors.
SUMMARY OF THE INVENTION The present invention provides a novel class of compounds, and pharmaceutically acceptable derivatives thereof, that are useful as inhibitors of aspartyl proteases, and in particular, HIV aspartyl protease. The compounds of this invention can be used alone or in combination with other therapeutic or prophylactic agents, such as anti-virals, antibiotics, immunomodulators or vaccines, for the treatment or prophylaxis of viral infection.
According to a preferred embodiment, the compounds 1 this invention are capable of inhibiting HIV viral replication in human CD 4 cells including Tcells, monocytic lines including macrophages and dendrocytes and other permissive cells. These compounds are useful as therapeutic and prophylactic agents to treat or prevent infection by HIV-1 and related viruses which may result in asymptomatic infection, AIDS-related complex acquired immunodeficiency syndrome or similar disease of the immune system.
It is a principal object of this invention to provide a novel class of compounds that are aspartyl AMENDED SHEET 5 protease inhibitors, and particularly, HIV aspartyl protease inhibitors. This novel class of compounds is represented by formula I: R\ R 1 R 7 5 YX R R
(I)
wherein each Z is R R7 (N-G R or or N r N ,Y
R
4 wherein any Z may be optionally fused with R6; each X and X' is independently selected from the group consisting of and -S(0)2; each Y and Y' is independently selected from the group consisting of -NR 2
-(C(R
2
>C=C(R
2 and -N(R 2
)-CH
2 each R 1 is independently selected from the group consisting of hydrogen; R6; C 1 -Cg alkyl; C 2
-C
6 alkenyl; C2-C6 alkynyl; C3-C6 cycloalkyl optionally fused with R C5-C6 cycloalkenyl optionally fused with R6; and where Rl's are attached to adjacent atoms, the R 's together with their attached adjacent atoms form a carbocyclic or heterocyclic ring system which may be optionally fused with R6; where any member of R 1 may be optionally substituted by one or more R2; AMENDED
SHEET
WO 97/27180 PCT/US97/01610 6 each R is independently selected from hydrogen; 3 R C1-C6 alkyl; C2-Cg alkenyl; C 2
-C
6 alkynyl; C 3
-C
6 cycloalkyl optionally fused with R6; C 5
-C
6 cycloalkenyl optionally fused with R 6 and where two R 2 's are attached to the same geminal atom, the R2's together with their attached geminal atom may form a spirocarbocyclic or spiroheterocyclic ring system; where any member of R 2 may be optionally substituted by one or more R; each R is independently selected from oxo, OR,.
N(R N(R ,N(R )-X-OR9, N(R 2
SR
9
X-
R9 O-X-N(R9)2, C(O)N(R halogen,
NO
2 CN, COOR 9 and R6; 4 each R is independently selected from from the group consisting of OR N(R X-R C(O)N(R )2 R6; C1-C6 alkyl; C2-C4 alkenyl; C3-C6 cycloalkyl optionally fused with R 6 C5-C 6 cycloalkenyl optionally fused with R where any member of R 4 may be optionally substituted by one or more groups independently selected from the group consisting of R 9 and R3; each R 5 is independently selected from the group consisting of H, OH, 0 and R 1 each R 6 is independently selected from the group consisting of aryl, carbocyclyl and heterocyclyl, wherein said aryl, carbocyclyl or heterocyclyl may be optionally substituted with one or more groups selected from the group consisting of oxo, -OR 9
-R
9
-N(R
9
(R
9 9 9 9 9 9 9 9 -N(R )-X-R 9
SR
9 -X-R -O-X-N(R 9 -R-OR
-CN,
-C0 2
R
9
-X-N(R
9
(R
9 halogen, -NO 2 and -CF 3 each R is independently selected from the group consisting of hydrogen, OH and 0; each R 8 is independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, carbocyclyl, and heterocyclyl; WO 97/27180 PCT/US97/01610 -7 each R 9 is independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, carbocyclyl, heterocyclyl, aralkyl, carbocyclylalkyl and heterocyclylalkyl wherein any aryl, carbocyclyl or heterocyclyl may be optionally fused with R 8 and wherein any member of R may be optionally substituted by one or more groups independently selected from the group consisting of -OR -N(R -CN, -NO 2
-X-R
8 N(R)2, -C(O)OR -N(R )-XNR 8 and halogen; each Q is independently selected from CH and N; each M is independently selected from the group consisting of NH, -NR 2 and each n is 1 or 2; each r is 0,1 or 2; each p is independently 1 or 2; each q is independently 1, 2 or 3; and each G is independently selected from the group consisting of
-NR
2 S(0) 2 and -C(R2)2-.
An alternate object of this invention is a novel class of compounds represented by formula IV: 7 R\
R
Y xN N R 4 (IV) R 1 wherein: X and X' are independently or Y is 2 -(C(R2) 2 )p or -N(R2)-
CH
2 and WO 97/27180 PCTIUS97/01610 8 each R, R R 7
R
4 p and M is independently as defined for formula I.
Another object of this invention is a novel class of compounds represented by formula V: 7 R 7 R R 11
(V)
wherein: X is or Y is 2 or -N(R2)-
CH
2
R
10 is 0 or H 2 each R11 is independently H, OH or 0, wherein both R 1 are not simultaneously hydrogen; Z is a structure of formula VI: [)qG
R
N J^ Q 8
R
R
4
(VI)
wherein any structure of formula VI is optionally fused with an aryl, carbocyclic or heterocyclic ring and is optionally substituted with 1-3 substituents independently selected from R2; and each R 1
R
2 R R 4 R p, q, G, M, Q and X' is independently as defined for formula I.
WO 97/27180 PCT/US97/01610 9 It is also an object of this invention to provide pharmaceutical compositions comprising the compounds of formulas I, IV and V and methods for their use as inhibitors of aspartyl protease, and particularly, HIV aspartyl protease.
It is a further object of this invention to provide methods for treating viral diseases, and in particular HIV-related diseases, using the compounds and compositions of this invention.
WO 97/27180 PCT/US97/Ej1610 DETAILED DESCRIPTIOaN OF THE INVENTION In order that the invention herein described may be more fully understood, the following detailed description is set forth. In the description, the following abbreviations are used: Designation Reagent or Fragment Ac acetyl.
Me methyl Et ethyl 0 Bn benzyl Trityl triphenylmethyl Asn D- or L-asparagine Ile D- or L-isoleucine Phe D- or L-phenylalanine Val D- or L-valine Boc tert-butoxycarbonyl Cbz benzyloxycarbonyl (carbobenzyloxy) RMOC 9 -fluorenylmethoxycarbonyl DCC dicyclohexylcarbodiimide DIC diisopropylcarbodiimide HO~t HOSu T FA DI EA
DBU
EtOAc t-Bu iBu
DMF
THP
THF
DMS 0 ethylcarbodiimide hydrochloride 1-hydroxybenzotriazole 1 -hydroxysuccinimide trifluoroacetic acid diisopropylethylamine 1, 8-diazabicyclo undec-7--ene ethyl acetate tert-butyl iso-butyl dime thyl f ormamide tertrahydropyran tetrahydrofuran dimethylsul foxide WO 97/27180 PCT/US97/01610 11 The following terms are employed herein: Unless expressly stated to the contrary, the terms "-SO 2 and 2 as used herein refer to a sulfone or sulfone derivative both appended groups linked to the and not a sulfinate ester.
The term "alkoxy" refers to an alkyl ether radical, wherein the term "alkyl" is as defined above.
Examples of suitable alkyl ether radicals include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tertbutoxy and the like.
The term "alkyl", alone or in combination with any other term, refers to a straight-chain or branch-chain saturated aliphatic hydrocarbon radical containing the specified number of carbon atoms, or where no number is specified, preferably from 1-10 and more preferably from 1-5 carbon atoms. Examples of alkyl radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, secbutyl, tert-butyl, pentyl, isoamyl, n-hexyl and the like.
The term "alkenyl", alone or in combination with any other term, refers to a straight-chain or branched-chain mono- or poly-unsaturated aliphatic hydrocarbon radical containing the specified number of carbon atoms, or where no number is specified, preferably from 2-10 carbon atoms and more preferably, from 2-6 carbon atoms. Examples of alkenyl radicals include, but are not limited to, ethenyl, E- and Z-propenyl, isopropenyl, E- and Z-butenyl, E- and Z-isobutenyl, E- and Z-pentenyl, E- and Z-hexenyl, Z,E- and Z,Z-hexadienyl and the like.
The term "anti-viral agent" or "antiretroviral agent" refers to a compound or drug which WO 97/27180 PCT/US97/01610 12 possesses viral inhibitory activity. Such agents include reverse transcriptase inhibitors (including nucleoside and non-nucleoside analogs) and protease inhibitors. Preferably the protease inhibitor is an HIV protease inhibitor. Examples of nucleoside analog reverse transcriptase inhibitors include, but are not limited to, zidovudine (AZT), dideoxycytidine (ddC), didanosine (ddl), stavudine (d4T), 3TC, 935U83, 1592U89 and 524W91. Examples of non-nucleoside analog reverse transcriptase inhibitor include, but are not limited to TIBO, delavirdine (U90) and nevirapine. Examples of HIV protease inhibitors include, but are not limited to VX-478 (Vertex, also known as 141W94 (Glaxo-Wellcome) and KVX-478 (Kissei)), saquinavir (Ro 31-8959, Roche), indinavir (L-735,524, Merck)), ritonavir (ABT 538, Abbott), nelfinavir (AG 1343, Agouron), palinavir (Bila 2011 BS), U-103017 (Upjohn), XM 412 (DuPont Merck), XM 450 (DuPont Merck), BMS 186318 (Bristol-Meyers Squibb), CPG 53,437 (Ciba Geigy), CPG 61,755 (Ciba Geigy), CPG 70,726 (Ciba Geigy), ABT 378 (Abbott), GS 3333 (Gilead Sciences), GS 3403 (Gilead Sciences), GS 4023 (Gilead Sciences), GS 4035 (Gilead Sciences), GS 4145 (Gilead Sciences), GS 4234 (Gilead Sciences), and GS 4263 (Gilead Sciences).
The term "aryl", alone or in combination with any other term, refers to a carbocyclic aromatic radical (such as phenyl or naphthyl) containing the specified number of carbon atoms, preferably from 6-14 carbon atoms, and more preferably from 6-10 carbon atoms. Examples of aryl radicals include, but are not limited to phenyl, naphthyl, indenyl, indanyl, azulenyl, fluorenyl, anthracenyl and the like.
The term "carbocycle" and "carbocyclyl" radical, refers to a non-aromatic stable 3- to 8- WO 97/27180 PCT/US97/01610 13 membered carbon ring which may be saturated, monounsaturated or poly-unsaturated. The carbocycle may be attached at any endocyclic carbon atom which results in a stable structure. Preferred carbocycles have 5-6 carbons.
The term "heterocycle" and "heterocyclyl" radical, unless otherwise defined herein, refers to a stable 3-7 membered monocyclic heterocyclic ring or 8- 11 membered bicyclic heterocyclic ring which is either saturated or unsaturated, and which may be optionally benzofused if monocyclic. Each heterocycle consists of one or more carbon atoms and from one to four heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. As used herein, the terms "nitrogen and sulfur heteroatoms" include any oxidized form of nitrogen and sulfur, and the quaternized form of any basic nitrogen. In addition, any ring nitrogen may be optionally substituted with a substituent
R
2 as defined herein for compounds of formula I. A heterocyclyl radical may be attached at any endocyclic carbon or heteroatom which results in the creation of a stable structure. Preferred heterocycles include 5-7 membered monocyclic heterocycles and 8-10 memebered bicyclic heterocycles. Preferred heterocycles defined above include, for example, benzimidazolyl, imidazolyl, imidazolinoyl, imidazolidinyl, quinolyl, isoquinolyl, indolyl, indazolyl, indazolinolyl, perhydropyridazyl, pyridazyl, pyridyl, pyrrolyl, pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrazinyl, quinoxolyl, piperidinyl, pyranyl, pyrazolinyl, piperazinyl, pyrimidinyl, pyridazinyl, morpholinyl, thiamorpholinyl, furyl, thienyl, triazolyl, thiazolyl, B-carbolinyl, tetrazolyl, thiazolidinyl, benzofuranoyl, thiamorpholinyl sulfone, oxazolyl, benzoxazolyl, oxopiperidinyl, oxopyrroldinyl, WO 97/27180 PCT/US97/01610 14 oxoazepinyl, azepinyl, isoxazolyl, isothiazolyl, furazanyl, tetrahydropyranyl, tetrahydrofuranyl, thiazolyl, thiadiazoyl, dioxolyl, dioxinyl, oxathiolyl, benzodioxolyl, dithiolyl, thiophenyl, tetrahydrothiophenyl and sulfolanyl, dioxanyl, dioxolanyl, tetrahydrofurodihydrofuranyl, tetrahydropyranodihydrofuranyl, dihydropyranyl, tetrahydrofurofuranyl and tetrahydropyranofuranyl.
The term "halogen" refers to a radical of fluorine, chlorine, bromine or iodine.
The terms "HIV protease" and "HIV aspartyl protease" are used interchangeably and refer to the aspartyl protease encoded by the human immunodeficiency virus type 1 or 2. In a preferred embodiment of this invention, these terms refer to the human immunodeficiency virus type 1 aspartyl protease.
The term "inert solvent" refers to a solvent reaction medium which allows the reagents to react together at a substantially increased rate relative to any reagent reacting with the designated solvent.
The term "leaving group" or "LG" refers to groups readily displaceable by a nucleophile, such as an amine, alcohol, phosphorous or thiol nucleophile or their respective anions. Such leaving groups are well known and include carboxylates, N-hydroxysuccinimide, N-hydroxybenzotriazole, halogen (halides), triflates, tosylates, mesylates, alkoxy, thioalkoxy, phosphinates, phosphonates and the like. Other potential nucleophiles include organometallic reagents known to those skilled in the art.
The term "protecting group" refers to a suitable chemical group which may be attached to a functional group and removed at a later stage to reveal the intact functional group. Examples of suitable protecting groups for various functional groups are WO 97/27180 PCT/US97/01610 15 described in T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis. 2d. Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); L. Paquette, ed. Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995).
The term "fused" whether preceded by the term "optionally" or not, refers to a structure wherein two distinct ring systems are joined together such that both rings share at least two common atoms. This can be envisioned as the replacement of a carbon-hydrogen or nitrogen-hydrogen bond on a ring atom with a carboncarbon (from a second ring) or nitrogen-carbon (from a second ring) bond. For example, a cyclohexyl ring fused to a second cyclohexyl ring results in a decahydronaphthalene, a cyclohexyl ring fused to a piperidine ring results in a decahydroquinoline or decahydroisoquinoline, or a phenyl ring fused to a thiazole ring results in a benzothiazole.
The term "substituted", whether preceded by the term "optionally" or not, and substitutions contained in formulas of this invention, refer to the replacement of one or more hydrogen radicals in a given structure with the radical of a specified substituent.
When more than one position in a given structure may be substituted with more than one substituent selected from a specified group, the substituents may be either the same or different at every position (for example, the moiety -N(R2) Typically, when a structure may be optionally substituted, 0-3 substitutions are preferred, and 0-1 substitutions is more preferred.
Most preferred substituents are those which enhance protease inhibitory activity or intracellular antiviral activity in permissive mammalian cells or immortalized mammalian cell lines, or which enhance deliverability WO97/27180 PCT/US97/01610 16 by enhancing solubility characteristics or enhancing pharmacokinetic or pharmacodynamic profiles as compared to the unsubstituted compound. Other more preferred substituents include those used in the compounds shown in Tables The term "pharmaceutically effective amount" refers to an amount effective in treating HIV infection in a patient either as monotherapy or in combination with other agents. The term "treating" as used herein refers to the alleviation of symptoms of a particular disorder in a patient or the improvement of an ascertainable measurement associated with a particular disorder. Specifically, with respect to HIV, effective treatment using the compounds and compositions of this invention would result in an improvement in an HIV associated ascertainable measurement. The term "prophylactically effective amount" refers to an amount effective in preventing HIV infection in a patient. As used herein, the term "patient" refers to a mammal, including a human.
The term "pharmaceutically acceptable carrier or adjuvant" refers to a carrier or adjuvant that may be administered to a patient, together with a compound of this invention, and which does not destroy the pharmacological activity thereof and is nontoxic when administered in doses sufficient to deliver a therapeutic amount of the antiretroviral agent.
As used herein, the compounds of this invention, including the compounds of formula I are defined to include pharmaceutically acceptable derivatives or prodrugs thereof. A "pharmaceutically acceptable derivative or prodrug" means any pharmaceutically acceptable salt, ester, salt of an ester, or other derivative of a compound of this WO 97/27180 PCT/US97/01610 17 invention which, upon administration to a recipient, is capable of providing (directly or indirectly) a compound of this invention or an inhibitorily active metabolite or residue thereof. Particularly favored derivatives and prodrugs are those that increase the bioavailability of the compounds of this invention when such compounds are administered to a mammal by allowing an orally administered compound to be more readily absorbed into the blood) or which enhance delivery of the parent compound to a biological compartment the brain or lymphatic system) relative to the parent species.
Pharmaceutically acceptable salts of the compounds of this invention include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acids include hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycollic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, ethanesulfonic, formic, benzoic, malonic, naphthalene- 2-sulfonic and benzenesulfonic acids. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts.
Salts derived from appropriate bases include alkali metal sodium), alkaline earth metal magnesium), ammonium and N-(C1- 4 alkyl) 4 salts.
The term "thiocarbamates" refers to compounds containing the functional group N-S0 2
-O.
The compounds of this invention contain one or more asymmetric carbon atoms and thus occur as WO 97/27180 PCT/US97/01610 18 racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers.
All such isomeric forms of these compounds are expressly included in the present invention. Each stereogenic carbon may be of the R or S configuration.
Although the specific compounds exemplified in this application may be depicted in a particular stereochemical configuration, compounds having either the opposite stereochemistry at any given chiral center or mixtures thereof are also envisioned.
Combinations of substituents and variables ,envisioned by this invention are only those that result in the formation of stable compounds. The term "stable", as used herein, refers to compounds which possess stability sufficient to allow manufacture and which maintains the integrity of the compound for a sufficient period of time to be useful for the purposes detailed herein therapeutic or prophylactic administration to a mammal or for use in affinity chromatography applications). Typically, such compounds are stable at a temperature of 400C or less, in the absence of moisture or other chemically reactive conditions, for at least a week.
The compounds of the present invention may be used in the form of salts derived from inorganic or organic acids. Included among such acid salts, for example, are the following: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2hydroxyethanesulfonate, lactate, maleate, WO 97/27180 PCT/US97/01610 19 methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate and undecanoate.
This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. The basic nitrogen can be quaternized with any agents known to those of ordinary skill in the art including, for example, lower alkyl halides, such as methyl, ethyl, propyl and butyl chloride, bromides and iodides; dialkyl sulfates including dimethyl, diethyl, dibutyl and diamyl sulfates; long chain halides such as decyl, lauryl, myristyl.and stearyl chlorides, bromides and iodides; and aralkyl halides including benzyl and phenethyl bromides. Water or oil-soluble or dispersible products may be obtained by such quaternization.
The compounds of this invention are those of formula I:
R
7
R
1 N R
(I)
wherein each Z is WO 97/27180 PCT/US97/01610 20
R
1
R
7
G
orR4 or or r N Y
/X'
R
wherein any Z may be optionally fused with R6; each X and X' is independently selected from the group consisting of and -S(0)2; each Y and Y' is independently selected from the group consisting of -(C(R2 -NR 2 >C=C(R2)2, and -N(R2)-CH2-; each R 1 is independently selected from the group consisting of hydrogen; R 6
C
1
-C
6 alkyl; C2-C 6 alkenyl; C2-C6 alkynyl; C 3
-C
6 cycloalkyl optionally fused with R C5-C6 cycloalkenyl optionally fused with R 6 and where R 's are attached to.adjacent atoms, the R 1 's together with their attached adjacent atoms form a carbocyclic or heterocyclic ring system which may be optionally fused with R 6 where any member of R 1 may be optionally substituted by one or more R2 each R 2 is independently selected from hydrogen; 3 R C1-C6 alkyl; C2-C 6 alkenyl; C2-C6 alkynyl;
C
3
-C
6 cycloalkyl optionally fused with R 6
C
5
-C
6 cycloalkenyl optionally fused with R and where two R 2 's are attached to the same geminal atom, the R2's together with their attached geminal atom may form a spirocarbocyclic or spiroheterocyclic ring system; where any member of R 2 may be optionally substituted by one or more R WO 97/27180 PCTIUS97/01610 21 39 each R is independently selected from oxo, OR, N(R9)2, N(Rg)-X-R 9 N(R )-XOR9 N(R9)-X-N(R 9 )2 SR X-
R
9 O-X-N(R )2 C(O)N(R 9 )2 halogen, NO 2 CN, COOR 9 and 6 R6; each R 4 is independently selected from from the group consisting of OR N(R9)2; XR9; C(O)N(R9)2;
R
6
C
1
-C
6 alkyl; C2-C4 alkenyl; C3-C6 cycloalkyl optionally fused with R C5-C 6 cycloalkenyl optionally fused with R where any member of R 4 may be optionally substituted by one or more groups independently selected from the group consisting of R 9 and R 3 each R 5 is independently selected from the group consisting of H, OH, 0 and R 1 each R 6 is independently selected from the group consisting of aryl, carbocyclyl and heterocyclyl, wherein said aryl, carbocyclyl or heterocyclyl may be optionally substituted with one or more groups selected from the group consisting of oxo, -OR 9
-R
9
-N(R
9
(R
9 -N(R SR9, -X-R -R -OR -CN,
-CO
2 R -X-N(R 9
(R
9 halogen, -NO 2 and -CF 3 each R 7 is independently selected from the group consisting of hydrogen, OH and 0; 8 each R is independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, carbocyclyl, and heterocyclyl; each R is independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, carbocyclyl, heterocyclyl, aralkyl, carbocyclylalkyl and heterocyclylalkyl wherein any aryl, carbocyclyl or heterocyclyl may be optionally fused with R 8 and wherein any member of R 8 may be optionally substituted by one or more groups independently selected from the grbup consisting of -OR -N(R8)2, -CN, -NO 2
-X-R
8
-X-
N(R -C(0)OR 8
-N(R)-XNR
8 and halogen; n WO 97/27180 PCT/US97/01610 22 each Q is independently selected from CH and N; each M is independently selected from the group consisting of NH, -NR 2 and each n is 1 or 2; each r is 0,1 or 2; each p is independently-1 or 2; each q is independently 1, 2 or 3; and each G is independently selected from the group consisting of
-NR
2 S(0)2, and -C(R 2 2 Except where expressly noted to the contrary, the term "[variable] as defined for formula I" refers to the definitions shown directly above. In addition, where no reference is made to a particular definition for a given variable, the definition is to be taken as that defined for formula I shown directly above.
Preferred compounds of formula I are those wherein each Y and Y' is independently selected from the group consisting of -NR2-, -(C(R22)p-M-, and -N(R2)-CH 2 and 3 9 each R is independently selected from oxo, OR N(R)2 N(R9)-X-R 9 N(R )-X-OR 9
SR
9
X-R
9 O-X-N(R )2 9 9 6 C(O)N(R halogen,
NO
2 CN, COOR 9 and R Alternate preferred compounds of formula
I
are those having the structure of formula IA:
R
7 5 R
(IA)
wherein each R 12 is independently selected from the group consisting of R 6
C-C
6 alkyl optionally substituted WO 97/27180 WO97/27180 PCT/US97/01610 23 with R6; C2-C6 alkenyl; C2-C6 alkynyl; C3-C6 cycloalkyl optionally fused with R C5-C6 cycloalkenyl optionally fused with R6; where any member of R 12 may be optionally substituted by one or more R 2 Preferred compounds of formula I are those wherein n is equal to 1; those having the structure of formula II:
(II)
and those having the structure of formula III: 7 7
R
R
R X Z RY __NR OHR xx
(III)
Also preferred are compounds according to formula I wherein X is or and Y is -(C(R 2 those wherein X is or -S(0)2and Y is those wherein X is or -S(0) 2 and Y-is -N(R 2 or -N(R 2 CH 2 WO97/27180 PCT/US97/01610 24 An alternate object of this invention is a novel class of compounds represented by formula IV:
(IV)
wherein: X and X' are independently or Y is -(C(R2) 2 or -N(R2)-
CH
2 and each R 1
R
2 R R 4 p and M is independently as defined for formula I.
Another object of this invention is a novel class of compounds represented by formula V:
(V)
wherein: X is or Y is -(C(R 2 2
-(C(R
2
-N(R
2 or -N(R2-
CH
2 R1 0 is 0 or H 2 each R1 is independently H, OH or 0, wherein both 1-1 R are not simultaneously hydrogen; Z is a structure of formula VI: WO 97/27180 PCT/US97/01610 25 R8
N
X'
R4
(VI)
wherein any structure of formula VI is optionally fused with an aryl, carbocyclic or heterocyclic ring and is optionally substituted with 1-3 substituents independently selected from R 2 (where in formula V, if 10 R is H2, a methylene is implied); and each R R R R, R, p, q, G, M, Q and X' is independently as defined for formula I.
Also preferred are those compounds having the structure of formula V, wherein
R
10 and R 1 1 are 0; compounds having the structure of formula V, wherein and R11 are 0; q is 1; G is S; and X' is compounds having the structure of formula V, wherein R0 and R1 are 0; q is 1; G is S; X' is and
R
4 is t-butylamino; compounds having the structure of formula V, wherein R and R1 are 0; X is Y is -(C(R2 and
R
7 is H; compounds having the structure of formula V wherein WO 97/27180 PCT/US97/01610 26- X and X' is Y is -(C(R 2 2
R
7 is H;
R
0 is H 2 and one R11 is H and one R 11 is OH; Also preferred are those compounds of formula V .wherein X and X' is Y is -(C(R 2 2
R
7 is H;
R
1 0 is H 2 one R11 is H and one R 11 is OH; and
R
2 within the definition of Y is selected from hydrogen, R 3 or C 1
-C
6 alkyl optionally substituted with R3; those compounds of formula V wherein X and X' is Y is -(C(R 2
R
7 is H;
R
1 0 is H 2 one R 11 is H and one R11 is OH; and
R
2 within the definition of Y is selected from hydrogen, -N(R or heterocyclyl, which may be optionally benzofused, and wherein said heterocyclyl may be optionally substituted with one or more groups selected from the group consisting of oxo, -OR 9 -R
(R
9 -N(R SR -X-R -O-X-N(R -R-
OR
9 -CN, -C02R 9
-X-N(R
9
(R
9 halogen, -NO 2 and -CF 3 those compounds of formula V wherein X and X' is Y is
R
7 is H;
R
1 0 is H 2 WO 97/27180 WO 9727180PCTIUS97/01610 -27 one R 11is H and one R 11is OH; and R 2within the definition of Y is selected :-rolm the group consisting of: 7 7 7 0
H
CH
3
N
H
0-b
I-$
3 C Me
N~KN<
'N N
H
7
N
N7
H
7 N- C-b
(N
H
WO 97/27180 PCT/US97/01610 28 CHSe
JNN
CH-
3 -7 'N 'N N
-A
I I 'N 'N N
-A
'N
42
NH
2 s N-_
OH
OH
S N- H2- HN2 WO 97/27180 PCT/US97/01610 29
OH
I N F F F F
F
those compounds according to formula V wherein: X and X' is Y is
R
7 is H;
R
1 0 is H2; one R11 is H and one R 11 is OH; and at least one R within the definition of Y is aryl optionally substituted with one or more groups selected from the group consisting of oxo,.-OR 9
-R
9
-N(R
9 (R9), -N(R SR -X-R9, -O-X-N(R9)2 -R -OR -CN,
-CO
2 R -X-N(R 9
(R
9 halogen, -NO 2 and -CF 3 those compounds according to formula V wherein: X and X' is Y is -(C(R 2 2
R
7 is H;
R
1 0 is H 2 one R11 is H and one R 11 is OH; and at least one R 2 within the definition of Y is Cl-C 6 alkyl optionally substituted with R3; those compounds according to formula V wherein: X and X' is Y is -(C(R 2 WO 97/27180 PCTIUS97/01610 30 R is H;
R
1 0 is H 2 one R 11 is H and one R 11 is OH; at least one R 2 within the definition of Y is CI-C6 alkyl optionally substituted with R 3 and 3 at least one R .within the definition of Y is pyridyl, triazolyl, oxazolyl, isoxazolyl, pyrimidyl, pyrazolyl, pyridazinyl, thiazolyl, imidazolyl, thienyl thiadiazolyl, oxadiazolyl, triazinyl or pyrazinyl wherein said R 3 may be optionally substituted with 1-3 substituents selected from -OR 9
-R
9
-N(R
9
)R
9 -N(R SR9, -X-R -O-X-N(R -R -OR -CN, -C0 2 R -X-N(R 9 halogen, -NO 2 and -CF 3 those compounds according to formula V wherein: X and X' is Y is
R
7 is H;
R
1 0 is H 2 one R 1 1 is H and one R 11 is OH; at least one R within the definition of Y is CI-C6 alkyl optionally substituted with R 3 and
R
3 within the definition of Y is aryl optionally substituted with 1-3 substituents selected from -OR 9 -R -N(R9)-X-R SR9, -X-R 9
-O-X-N(R
9 2 -R -OR 9 -CN, -C0 2
R
9
-X-N(R
9
(R
9 halogen,
-NO
2 and
-CF
3 Also preferred are those compounds according to any of the aforementioned preferred compounds of formula V wherein:
R
1 is benzyl; and Z is WO 97/27180 PCT/US97/01610 31
N
H
those compounds according to any of the aforementioned preferred compounds of formula V wherein: R is benzyl optionally substituted with 1-3 substituents selected from -OR 9 SR -X-R, 9 9 -R -OR -CN, halogen,
-NO
2 and -CF 3 those compounds according to any of the aforementioned preferred compounds of formula V wherein:
R
1 is benzyl optionally substituted with 1-3 substituents selected from -OR -N(R 9
(R
9 SR -X-R, -R -OR -CN, halogen, -NO 2 and -CF 3 and Z is O N
H
those compounds according to any of the aforementioned preferred compounds of formula V wherein R 1 is benzyl optionally substituted with 1-3 substituents selected from the group consisting of OCH 3 OH and NH 2 WO 97/27180 PCT/US97/01610 32 those compounds according to any of the aforementioned preferred compounds of formula V wherein R 1 is benzyl optionally substituted with 1-3 substituents selected from the group consisting of OCH 3 OH and NH 2 and wherein Z is
N
O N<
H
An alternate embodiment of this invention is compounds according to formula V, wherein: 7 R R R R 11 R"
(V)
each R 6 is independently selected from the group consisting of aryl, carbocyclyl and heterocyclyl, wherein said aryl, carbocyclyl or heterocyclyl is optionally substituted with one or more groups selected from the group consisting of oxo, -OR 9
-R
9
-N(R
9
(R
9 -N(R SR -O-X-N(R9)2, -R -OR 9
-CN,
-C0 2
R
9
-X-N(R
9
(R
9 halogen, -NO 2
-CF
3
-O-(CH
2 )q-R 6
-O-(CH
2 )q-OR 2,3-methylenedioxy and 3,4methylenedioxy; and each X, Y, Z, R 1
R
2
R
3 R R R R R Q, M, n, r, p, q and G is independently as defined for formula I; and WO 97/27180 PCTIUS97/01610 33 those compounds according to formula V, wherein: each R is independently selected from the group consisting of aryl, carbocyclyl and heterocyclyl, wherein said aryl, carbocyclyl or heterocyclyl is optionally substituted with one or more groups selected from the group consisting of oxo, -OR 9 -R -N(R 9
(R
9 -N(R )-X-R 9 SR -X-R 9
-O-X-N(R
9 2 -R9-OR -CN, 9 9 9 6 -C0 2 R -X-N(R 9
(R
9 halogen, -NO 2
-CF
3 -O-(CH2)q-R 9
-O-(CH
2 )q-OR 2,3-methylenedioxy and 3,4methylenedioxy;
R
2 within the definition of Y is selected from hydrogen, R or C 1
-C
6 alkyl optionally substituted with R and each X, Y, Z, R 1
R
3
R
4
R
5 R R, R Q, M, n, r, p, q and G is independently as defined for formula I.
those compounds of formula V wherein X and X' is Y is -N(R2)
R
7 is H; is H 2 and one R11 is H and one R 11 is OH; and those compounds of formula V wherein X and X' is Y is -(C(R 2 M is O;
R
7 is H; R1 0 is H 2 and one R 11 is H and one R 11 is OH.
Also preferred is the compound of formula I having the structure of formula IX: WO 97/27180 PCT/US97/01610 34 7 R\ RR
NR
7 x- H R 1
.(IX)
wherein X is or -S(0) 2 and the compounds of formula IX wherein X is Y is -(C(R2) 2 and R7 is H; and those compounds of formula IX wherein X is Y is -N(R 2 and
R
7 is H; and those compounds of formula IX wherein X is Y is and R 7 is H.
Also preferred are those compounds of formula I having the structure of formula XII: R R R X H R 1
X
(XII)
wherein X and X' are independently or those compounds of formula I having the structure of formula XII, wherein X and X' are independently or -S(0) 2 and WO 97/27180 PCT/US97/01610 35 R is l-amino-2-hydroxyindanyl; and compounds of formula I having the structure of formula XII, wherein R 4 is l(S)-amino-2(R)-hydroxyindanyl.
Also preferred are the compounds according to formula I, having the structure of formula XIII:
R
7
R
1 R H R H X R9
XIII)
wherein X and X' are independently or compounds according formula I having the structure of formula XIII, wherein X is or X' is Y is -(C(R 2 2 or -N(R 2 and
R
7 is H; compounds of formula I having the structure of formula XIII, wherein X is X' is Y is -(C(R 2 and
R
7 is H; those compounds of formula XIII wherein X is X' is Y is 2
R
7 is H; and WO 97/27180 PCT/US97/01610 36
R
2 within the definition of Y is selected from hydrogen,
R
3 or C1-C6 alkyl optionally substituted with R3 those compounds according to formula XIII wherein: X is X' is Y is -(C(R 2 2 R is H; and
R
2 within the definition of Y is selected from hydrogen, -N(R or heterocyclyl, which may be optionally benzofused, and wherein said heterocyclyl ,may be optionally substituted with 1-3 groups selected from the group consisting of oxo, -OR 9
-R
9
-N(R
9 (R -N(R SR -X-R -O-X-N(R 2 -R -OR -CN, -C0 2
R
9
-X-N(R
9
(R
9 halogen, -NO 2 and -CF 3 those compounds according to formula XIII wherein: X is X' is Y is -(C(R 2 2
R
7 is H; and 2 at least one R 2 within the definition of Y is selected from the group consisting of: OH N
H
K
3 C Me
K)
:-l zIm 6\-,Z z z \z T 6 z M t\x 6 6 \tz-: d\-x WO 97/27180 PCTIUS97/01610 38 Nt4 S N-
OH
OH
s N.
N-_
04n
N-
OH
F &F F &F those compounds according to formula XIII wherein: X is X, is Y is 2 R is and WO 97/27180 PCT/US97/01610 39 at least one R 2 within the definition of Y is aryl optionally substituted withone or more groups selected from the group consisting of oxo, -OR 9
-R
9
-N(R
9 (R9), -N(R SR -X-R -O-X-N(R 9 -R -OR -CN, -C0 2 R -X-N(R 9
(R
9 halogen,
-NO
2 and -CF 3 those compounds according to formula XIII wherein: X is X' is Y is -(C(R 2 2
R
7 is H; and at least one R 2 within the definition of Y is C*-C6 alkyl optionally substituted with R3 those compounds according to formula XIII wherein: X is X' is Y is 2) R is H; and at least one R 3 within the definition of Y is pyridyl, triazolyl, oxazolyl, isoxazolyl, pyrimidyl, pyrazolyl, pyridazinyl, thiazolyl, imidazolyl, thienyl thiadiazolyl, oxadiazolyl, triazinyl or pyrazinyl wherein said R 3 may be optionally substituted with 1-3 substituents selected from -OR, -R 9 -N(R -N(R SR9, -X-R -O-X-N(R9)2, -R -OR, -CN,
-CO
2 R -X-N(R 9 halogen,
-NO
2 or -CF 3 those compounds according to formula XIII wherein: X is X' is Y is
R
7 is H; and R within the definition of Y is aryl optionally substituted with 1-3 substituents selected from -OR 9 -R
(R
9
-N(R
9
)-X-R
9
SR
9 -X-R -O-X-N(R 9 2 WO 97/27180 PCT/US97/01610 40 -R -OR 9 -CN, -C0 2
R
9 -X-N(R9) halogen,
-NO
2 or
-CF
3 those compounds according to any of the aforementioned preferred compounds of formula XIII wherein: each R 1 is benzyl; and 9 each R not within the definition of Y is 2hydroxyindanyl.
those compounds according to any of the aforementioned preferred compounds of formula XIII wherein: each R 1 is independently selected from benzyl optionally substituted with 1-3 substituents selected 9 9 9 9 9 9 9 from -OR 9
SR
9 -R -OR, -CN, halogen,
-NO
2 and -CF 3 those compounds according to any of the aforementioned preferred compounds of formula XIII wherein: each R 1 is independently selected from benzyl optionally substituted with 1-3 substituents selected 9 9 9 9 9 9- 9 from -OR (R SR 9
-X-R
9 -R OR, -CN, halogen, -NO 2 and -CF 3 and each R 9 not within the definition of Y is 2hydroxyindanyl; those compounds according to any of the aforementioned preferred compounds wherein: each R1 is independently selected from benzyl optionally substituted with 1-3 substituents selected from the group consisting of OCH 3 OH and NH 2 and those compounds according to any of the aforementioned preferred compounds wherein: each R1 is independently selected from benzyl optionally substituted with 1-3 substituents selected from the group consisting of OCH3, OH and NH 2 9 each R not within the definition of Y is 2hydroxyindanyl.
WO 97/27180 PCT/US97/01610 41 Another embodiment is compounds according to formula XIII, wherein: 7 RY R
R
7 X R 9 7LX( 9" P
(XIII)
each R is independently selected from the group consisting of aryl, carbocyclyl and heterocyclyl, wherein said aryl, carbocyclyl or heterocyclyl is optionally substituted with one or more groups selected from the group consisting of oxo, -OR 9
-R
9
-N(R
9
(R
9 -N(R SR 9 -X-R -O-X-N(R 9 -R -OR9, -CN, -CO2 R 9
-X-N(R
9
(R
9 halogen, -NO 2
-CF
3
-O-(CH
2 )q-R 6
-O-(CH
2 q
OR
9 2,3-methylenedioxy and 3,4methylenedioxy; and each X, Y, Z, R R 2
R
3 R R R R R, Q, M, n, r, p, q and G is independently as defined for formula XIII.
Another embodiment is compounds according to formula XIII, wherein: R R' R .4 x N R9
XIII)
wherein R 2 within the definition of Y is selected from hydrogen, R 3 or C 1
-C
6 alkyl optionally substituted with R3 WO 97/27180 PCT/US97/01610 42 each R 6 is independently selected from the group consisting of aryl, carbocyclyl and heterocyclyl, wherein said aryl, carbocyclyl or heterocyclyl is optionally substituted with one or more groups selected from the group consisting of oxo, -OR 9 -N(R )-X-R 9
SR
9 -X-R -O-X-N(R 9 2 -R -OR -CN, -C0 2 R -X-N(R 9
(R
9 halogen, -NO 2
-CF
3 -O-(CH2)q-R
-O-(CH
2 )q-OR 2 ,3-methylenedioxy and 3,4methylenedioxy; and each X, Y, Z, R 1 R3, R R5, R R R Q, M, n, r, p, q and G is independently as defined for formula XIII.
Another embodiment is compounds of formula I having the structure of formula XIII, wherein X is X' is Y is -N(R 2 and
R
7 is H; compounds of formula I having the structure of formula XIII, wherein X is -SO 2 X' is Y is -(C(R 2 2 and R is H; and compounds of formula I having the structure of formula XIII, wherein X is -SO 2 X' is Y is and
R
7 is H.
In an alternate embodiment, preferred compounds are those of formula V wherein is H 2 and one R 11 is H and one R 1 1 is OH; and WO 97/27180 PCT/US97/01610 43 Z is selected from the group consisting of:
R
2 NR and N O) NHtBu O J NHtBu and R 2 is as defined in formula I; and those of formula V wherein Z is selected from the group consisting of H H H H N and N O J NHtBu O "NHtBu is H 2 and one R11 is H and one R 11 is OH.
Also preferred are those compounds of formula V wherein X and X' is Y is -(C(R 2 2
R
7 is H; is H 2 and one R 11 is H and one R11 is OH; and those compounds of formula V wherein X and X' is Y is -N(R2) R is H;
R
10 is H 2 and WO 97/27180 PCT/US97/01610 44 one R11 is H and one R11 is OH, and those compounds of formula V, wherein X and X' is Y is -(C(R2) 2 M is O;
R
7 is H; R1 0 is H 2 and one R 11 is H and one R 1 1 is OH, and the aforementioned compounds of formula V wherein Z is selected from the group consisting of:
R
2 2 rN
N
Y and
"N
O ^NHtBu O NHtBu 2 and R is as defined in claim 1.
Also preferred are those compounds of formula V wherein X and X' is Y is -(C(R 2
R
7 is H; R1 0 is H 2 and one R 1 is H and one R11 is OH; and those compounds of formula V wherein X and X' is Y is
R
7 is H;
R
10 is H 2 and one R 11 is H and one R 1 1 is OH, and those compounds of formula V, wherein X and X' is Y is -(C(R 2 2 WO 97/27180 PCT/US97/01610 45 M is O;
R
7 is H; R is H 2 and one R 1 is H and one R 11 is OH, and the aforementioned compounds of formula V wherein Z is selected from the group consisting of: H H
"H
N and N 0 NHtBu 0- J NHtBu Also preferred are compounds of formula I wherein: (1) Z is selected from the group consisting of -X'R -N(R 4 and formula VI;
(VI)
WO 97/27180 PCT/US97/01610 46 wherein any structure of formula VI is optionally fused with an aryl, carbocyclic or heterocyclic ring and is optionally substituted with 1-3 members independently 2 selected from R and 1 2 3 4 5 6 7 8 9 each X, Y, Y' R 1
R
2
R
3 R R5, R 6 R, R R Q, M, n, r, p, q and G is independently as defined in for formula I.
Another embodiment of this invention relates to the process for preparing a compound of formula XIV: R1 R6NH
XIV
wherein R 1 and R 6 are.defined as in formula I, comprising the steps of: reacting a compound of formula XV: R1 NBoc
O
XV
wherein R 1 is defined as in formula I, in an inert solvent, preferably an ethereal solvent such as diethyl ether or THF, with a base, preferably an alkali metal amide such as lithiumdiisopropylamide at a temperature between about -78 °C to about 25 0C; reacting the product of step with an aldehyde R CHO followed by an optional treatment with a dehyrating agent, preferably Martin's sulfurane WO 97/27180 PCT/US97/01610 47 dehydrating agent, wherein R6 is defined as in formula I to give a compound of formula XVI: R1 R6 o NBoc O 0
XVI
wherein R 1 and R are defined as in formula I; reacting the product of step in an inert solvent, preferably methanol, with hydrogen gas in the presence of an hydrogenation catalyst, preferably palladium on carbon, followed by treatment with an anhydrous acid, preferably trifluoroacetic acid or 4N HC1 in dioxane to give a product of formula XIV.
Another embodiment of this invention relates to the process for preparing a compound of formula XVII: R1 R f N
XVII
1 2 wherein R 1 and R 2 are defined as in formula I, comprising the steps of: reacting a compound of formula XVIII: WO 97/27180 PCT/US97/01610 48 R1
NH
R2 0 O
XVIII
wherein R 1 and R 2 are as defined in formula I, in an inert solvent-, preferably DMF or THF, with a base preferably sodium hydride, then bromomethylacrylic acid at a temperature between about -78 °C to about 25 oC; reacting the product of step with an oxidizing agent, preferably ozone and if necessary a reductive work-up with a reducing agent such as dimethylsulfide; reacting the-product of step in an inert solvent, such as DMF, with thioproline t-butylamide and suitable amide-bond coupling reagents, preferably EDC, HOBT and N-methylmorpholine, to give a product of formula XVII.
Another embodiment of this invention relates to the process for preparing a compound of formula XIX:
OY'
R 1
SNH
r 0
XIX
wherein R and r are defined as in formula I, comprising the steps of: WO 97/27180 PCT/US97/01610 49 reacting a compound of formula XX 0 R 1
NPG
xx wherein R is defined as in formula I and PG is a Nprotecting group, such as those described in Greene and Wuts (infra), preferably p-methoxybenzyl, an inert 'solvent, preferably THF, with a base, preferably lithiumdiisopropylamide at between about -78 oC to about 25 then a bis-leaving group alkane of formula
XXI:
L
LG
XXI
wherein LG is selected from halo, preferably chloro or iodo, arylsulfonate esters, preferably tosyl, and alkylsulfonate esters, preferably mesyl, and r is defined as in formula I, to give a product of formula
XXII:
R 1
NPG
LG r 0
XXII
wherein R and PG are defined as in formula XX and LG and r are defined as in formula XXI; reacting the product of step in an inert solvent, preferably THF, with a base, preferably WO 97/27180 PCT/US97/01610 50 lithiumdiisopropylamide, at between about -78 °C to about 25 °C to give a product of formula XXIII: D1
XXIII
wherein R 1 is defined as in formula I and PG is a Nprotecting group; reacting the product of step in an inert solvent with a reagent suitable for removal of the Nprotecting group PG, such as those described in Greene and Wuts (infra), to give a compound of formula XIX.
In another embodiment, compounds of formula I with structures VII, VIII, IX, and X are preferred:
R
5
R
1 R R 1 Y XN N .NR 4 (VIII RHR7 7 Y ,XN .LNR 4
(IX)
7t R 7_ OH V hR7 Y'XN W-I--N .X" n
(X)
where all definitions of variables for formula I apply.
Preferred R groups for formula I include: C1-C6 alkyl and alkenyl optionally substituted with R6; 2 where two R taken together form a spriocyclic ring and
C
3 -Cg cycloalkyl or cycloalkenyl optionally fused with
R
6
R.
PCT/tJS97/01610 Vertex Pharmaceuticals Inc., et a! Our Ref.: B 2555 POT VOSSiUJS PARTNER PATENq 1A WAALTE 31 8-ry 4 3175 ML0)i6HE f Fe. 1,998 5;1 Preferred compounds of this invention of formula I include the specific compounds contained in Tables TABL 1
OH
/N 0 k4 AMENDED S-E'ET 52 AMENDED SHEET 53 is Ph 0 0 0 16 Ph 0 00 17 Ph H3C e 18q Phpri~ Et aN 0 00 19 Ph HO IC N 'sAY 0 o" 21 Ph .1 15,a0Me
H
2 N" N 0 00 22 PhCM 0 e
H
3 C fl JN
N
H 0 00b 23 ph H 00_ q AME1~4DEu SliEET 54 24 Ph 0 0 0 Ph AcO Ac0-mI 26 P h 0 27 Ph N ci 0 00" 2 7 P Ih m 0 29 Ph1C Pr 0 00 29P Ph 0 00 wEtAoo sa 55 HN> P~rOMe
H
3
CO
2 C A N- N 1 0 00 HN OmN
H
3 C2 N
N-K.
0 0 0 HN O0m e CO KNf...C% 38( Phi 0M H00 H HNh (or, 0Me N N 0 00 Ph N re
NN
001 AMENDED SHIEET 56 42 -Ph OMe o 0 43 Ph 0 &0 0 NN 0 0 4 Ph O2N N OM e 0 0 46 Ph 47 Ph o is 0 e 48 Ph N I 1 0' A~o 0 00 NHtBu 00
H
0 0 Ht~u AMENDED S.EET 57 51 h <NIN P h 0 NHt~u 0 52 P h H.
H
2 N
.N
0 0 NHt~u H2N 0 0 NHt~u 0> 0 NHt~u
H
H
H
0 NHtBu 56
H
57 00 NHtSu
L~
0 53 58
H,
H
HO 0 N~tSu 59 0 NHtBu
N
N0 NHtu 62 o NHtu 63
NH..
O, NHIBu AMENDED
SKET
59 64H,
"H
NHN
N--0 NHt~u 66 a NHt~u 67
"H
HN 0 NHtBu 68H,
"H
INN N 0 NHtBu 69 O 0 N HtMu PAMENO4DE
SHEET
60
OKN
Etoi- 0 NHtBu 71 N 0 NHtBu
H
72 0
H..
H
0 NHtBu AMENDED SHEET 61 76
H
o NHt~u 77 123
H,
'H
s _N_0 NHt~u 1243.
N
H
0 NHt~u
HH
125 0 NHtBu 125 ~:1 0 NHt~u rln O< AMENDED SHEET 62- 127
H
o NHtSu 129
H,
N "H o NHt~u
N
H
N-0 0 NHt~u AMENDED SHEET 63 133 N.
H,
N H 0 NH t~u 135 N.
H,
0 NHtBu
N
No 0 NHt~u 1386II
H
2
H
N0 NHtBu ivtN4OED SHEECT 64 139 1 lo I r-1 140
-H
o NHtBu 141
OH,
'H
0 NHtBu 1442~N~A% 0 NHt~u AENDED SHEET 9 9 145 146
.NNH
N 147 N SJi
N
NH
2 00b 148 Pri'i 0 N00 150
NN
""J"NDED SHEET i~N~ED SHEEr 66 155 I\ 156 0 hSS'EET I.33HS 030N3RV ~HN0N z091 NR T
"N.
BT
vw-s 091 0
N
L9 68 163 No 164
N'F
0 165 QyH
NN
166
NN
168 (3 N 2 AiVENID ED SHEET 99
N--
S-i
N
AMENDED ShEETf 70 179 cI\ 180 AMENDED SHEET 71. 182.i 0 182 Nz 183 184
N
N-
185
NZ
a AAviENDED SHEET 72 AMIENDED SHEET 73 1 9 3
NH
N
194
NH
2
N-N
195
NH
2
N
196 0-~NH 2 198, 197 NH2 198 NH2 lz o C. rE'F~iET 74 199
NH
NH
HO'
NN
2010~FH 202
NN
204 Hz 2 NH2
NN
9i 75 260
/N-N
N
299 ~~1 AMENDED
SHEET
76 300 11K 301.H, N
H
0 NHtBu 302H.
H
0 NHt~u Me 303 KAK 'H o NHt~u 304
M
iiiiI~i0 NHtBu 305
F
3 0 NHtBu AMENDED HT 77
H.
H
0 NHt~u
H,
H
0 NHtBu
H.,
H
0 NHt~u AMECNDED CI 2
ECT
78 1 1 312
H.
H
0 NHt~u 313 I
N
314 0 NHt~u Me 315 F H..
OMe
H
MeO N'0 NHtBu
N
317
HH
N0 NHt8u Me ANIEEIDED SHEET 0 79 80 324 BnO 'N H 0 NHt~u 325
H,
'N N o NHtBu 326MOH.
NN H N0 NHtBu 327 o NHtBu f/VAT 0 AMENDED SHEET t 81. TABLT~r
OHR
cmpd. A R Z NO 78 -Ph HOH
H
Bn N H -Ph H OH N ]Bn 83 I-Ph EnHOH 00 84 Ph Bn OH
QJH
MUOED Si.OEET 82 1. j 4! ~r dI/s NOODIED 19AVE, 83 91 Bn O 00 92 -Bn O
NN
93 B3nO H
OH,,
0 94 Bi H O H OH
N>(N
Oil 96 Bn OH Am~tgjoS1VLEEI 84 208 H9 BnH MeOT 209 H En 22.0 210
OH
212.zN.Bn
OH
-f
H
H
0 0 211 Bn HOH 2132-. Bn OH
HH
0 0 0 0 AMENDED T 85 215 Sri H OH
NN
215 Bni H 9H 0)7 217 BiH OH 00 219 Bri OH 00
SHEET
86 220 n H OH
N-
222 Bn
HOH
,N-
224 B~Il n
OH
OH
225 Bn H OH AMENDED SHEET 87 C. AMENDED SHEET S 88 232 Bri OH MeOY 233 Bn O 234 -5-.Bn OH 235 Bn HOH
F
3 C H c y 237 Bn HO MeH AMENDED SHE-ET *9 S.
89 AMENOED SH-EET 90 V AMENDED SHEET 91 250 En HOH
H
2
H
251. Bi H NyN,
HO
252 En Q 253 S H OH 254 En O H O
N-
I
AIMENDED
SHEET
S 0 92 AMENDED SV-EET S e 93 266 A Bn OH
NN
Me 267 A Bn OH
H
0 8 268 Bn OH
H
N M 0 269 Sn OH
H
F O 270 Bn OH H O 271. Bn H OH
N
Me AMENDED SHEET 94 272 7Bn
H
CF
3
N.
03
N.O
273 7 Bn HOH
NH
NN
274 7 n HOH
NH
700 277 7 n H OH
N
AMENDED SHl-EET 95 278 En
HOH
NFH
Me 279 n H OH OMe 0N yN, MeO.
NH2 H
O
NN
N0 H 0
N
Me 282 n H OH H N,'8
N-N
N
F
283 n H OH 00 A mENOE StA.EET -96 284 Bn H 0~0 285 Bn H 0N, 286 En HOH N ~N.
287 HO nHOH
NN
NY
289 BO> n HOH
NN
AMENDED S 4.EET 97 AMtNOC~sAE 98
TABLLRI
OH
A Z 0 Cmpd No. A Z 97 Fp-Ph 0 0 NI~t~u 0 00 NHtBu AMErNDED S)l!ET 99 99-P h 0 0 NHtBu 100 Ph Ph 0N Nt~ 101 K r- N"Z ""Ph r- NN oN 00 NHtBu 102 P h Ph '0 NHtBu 0 103 Ph s" 0 NHt~u 00 TAaLz&A 0 A -I z 0 cmpd No. A z 104 ,hr-s
N
~N N y 0 NHtBu 0 m p(I AMENDED SHiEET 100- 105 HN ""Ph 00 ONMt u 106 -IPh 0 0 NHtBu 107 Ph 0 NHtBu 00 NHtBu 109 hX P h 0 NHtBu 0 110 Ph 0 NHt~u 0 NHtBu 112 7- 0 NHtSu 10o1 258 AMENDED
SHEET
102 cmpd No. A
R
1 z hG -Ph
H
E3n N0 0 11.7 HN>."'Ph
H
N Bn ,N 0 0 0 118 Ph
H
Bn eN y0 0o 0 119 RPh 13n H HN')"h .NOy 120 ]Ph n H 121 -,Ph Bn H 122 Ph Bfl H I ON 0 00 0 WO 97/27180 PCT/US97/01610 103 The preferred compounds of this invention are compound numbers (as in Tables 1, 2, 3, 4, 7, 8, 9, 13, 14, 16, 17, 18, 20, 23, 24, 25, 26, 32, 35, 38, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 62, 63, 72, 75, 76, 78, 80, 82, 83, 91, 92, 94, 95, 96, 101, 102, 109, 121, 122, 123, 124, 126, 127, 128, 129, 131, 132, 133, 134, 135, 137, 138, 140, 141, 145, 146, 147, 149, 150, 155, 156, 160, 161, 162, 164, 165, 170, 171, 175, 176, 177, 179, 180, 185, 186, 190, 191, 192, 194, 195, 200, 201, 208, 219, 220, 228 and 264. More preferred are compound numbers: 2, 7, 8, 9, 14, 18, 20, 25, 26, 32, 38, 45, 47, 48, 49, 50, 51, 53, 54, 62, 63, 72, 82, 83, 91, 92, 94, 95, 96, 123, 126, 140, 141, 219, 220, 228 and 264. Even more preferred are compound numbers: 7, 8, 9, 20, 45, 50, 51, 53, 54, 82, 83, 92, 94, 96, 219, 220, 228 and 264.
In an alternate embodiment, this invention also relates to novel methods for preparing compounds and intermediates of the following structures.
One embodiment relates to a process The compounds of this invention may be synthesized using conventional techniques.
Advantageously, these compounds are conveniently synthesized from readily available starting materials.
Although the syntheses of the compounds of this invention are known to those of skill in the art, the following general schemes are set forth to illustrate these methods. These schemes should not be viewed as limiting the scope of this invention in any way.
Using standard techniques, compounds of the present invention having the general formula I may be obtained as described in the following schemes: WO 97/27180 PCT/US97/01610 104 SCHEME I
P
H 0 2 2 0 reductive amniation RH 0 H R
R
El 1. doprotect I2. base heat
CQ
2 Me Ph 3 P .N -K-%-C0 2 Me
H
EV
IMg MeCH
HN
R
0 Ell $i1. protect 0 El
-OH
1. 0-activation 2. nucleophile 1 Boc 2 O, DMAP 2. base, "E" 3. deprotect E 9I
H
0 E3 WO 97/27180 PTU9I11 PCTIUS97/01610 105 SCHEME I (cont'd) P -NJyOH H o) H o 1. ethyidethyi phosphoranyl methanesulfonate; n-BuUl 2. reduction PN Jl-S Z M H 0 0 deprotect 2. cyclize N H 0 0; $.0
EIV
i1. M*ONHMe EDCI 2. reduce 3. TMSCH 2 MgBr 4. BF 3 ettierate 5. AcSH liv 6. C12 ACOH HCI Ellb 1. deprotect I2. cychize 1 protect 2. base, "E" 3. deprotect RQ I WO 97/27180 PCTIUS97/01610 106 SCHEME 2 H o 1. R 2
NH
2 2. deprotection 3. reduction 1. R 2
NH
2 R 2. deprotection 3. reduction R NH NH P.N. ),YOH H 0 oxallyi chloride or SOlm 2
R'
R N NH 0 RN. NH
ES
0
NH
0 WO 97/27180 PCT/U.S97/01610 107 SCHEME 3 0
R
2 R 2
H
Route A Rl 0
EIX
j1. deprotect 2. base /Iheat R2TR R2
NH
0 E7 t1. deprotect 2. base heat J.N TBDMS
EX
0 0- activationR p.N
'IO
base
H
Route B Route C 1. TBDMSCI 0 2. deprotect Br 1 3. base, R 2 AOMe R R
H
WO 97/27180 PCTIUS97/01610 108 SCHEME IV 1. NaH 0 z2. deprotecP, (Z El E7)
OH
ES
R12
H
N~ N OR 1 0 Ph EXIb Ph
EXI
0 C02NHtBu EXIc WO 97/27180 PCT/US97/01610 109 SCHEME S 1. NaH 1. allyl bromide (Z =E1 E6) 0 z 2.epox e 2 c l
EXII
HN(R
2 2 OH
R
2 Z E R 1. (R 1
)NH
2 2. R 4 X-Act OH R z -XR' E9 (X C(0) or SO SCHEME 6
R
1 1. O -protect OH 2. base 'E N R 12 3. deprotect 0
EXIV
R
1 E0 E11 Methods for producing the compounds of this invention are well known in the art of organic synthesis. Several intermediates are commercially available, e.g. from Aldrich Chemical Company, Inc., Milwaukee, WI. The synthesis of heterocycles E1-E6 (Schemes 1 and 2) begins with any protected amino aldehyde, the preparations for which are well known in the art from suitably protected amino acids, esters or alcohols. In the case of the this intermediate, transient protection of the amino group may be accomplished by means known in the art (see, e.g. T.W.
Greene and P.G.M. Wuts "Protective Groups in Organic Synthesis", Second Edition, pp. 309-405 ©1991 John Wiley and Sons, Inc. New York, NY and E. Gross and J.
WO 97/27180 PCT/US97/01610 110 Meinhofer "The Peptides, Vol. 3: Protection of Functional Groups in Peptide Synthesis" pp. 3-88; ©1981 Academic Press, Inc. New York, NY). Carbamates such as Boc, Fmoc, Alloc and Cbz are particularly convenient protecting groups, the introduction and removal of are described in the above references.
The synthesis of El is illustrated in Scheme 1. The protected amino aldehyde is treated with an alpha substituted or alpha, alpha disubstituted amino ester under typical reductive amination conditions well known in the art, such as sodium cyanoborohydride in a .solvent mixture of DMF/Acetic acid. The resulting compound El is then deprotected and free based with either a tertiary amine base or potasium carbonate in methanol to effect cyclization to form EII The resulting secondary amine may the be protected with groups (detailed in the references above) such as benzyl or t-butyloxycarbonyl (Boc) utilizing conditions well known in the art to form analogs of El.
Preparation of E2 is achieved by reaction of a starting aldehyde with ethyl diethylphosphoranylmethanesulfonate and subsequent reduction of the double bond (see: Gennari et al., Angew. Chem. Int. Ed. Engl., 33, pp. 2067-69 (1994)) to yield compound EIIIa. Cyclization may then be achieved by deesterification and activation of the sulfonate moiety as described in Gennari, followed by deprotection of the nitrogen protection group to yield the cyclized product EIV. Alternatively, an amino acid may be converted to compound EIIIb using standard synthetic methods illustrated in Scheme 1. Compound EIIIb can be cyclized to afford compound EIV. Compound EIV may then be N-protected, for example, in the presence of Boc anhydride and DMAP (see: Flynn et al.,
I
WO 97/27180 PCT/US97/01610 111 rg. cnem. 48, pp. 2424-26 (1983)), and treated with a non-nucleophilic base such as LDA or hexamethyldisilazane to generate the anion at the center alpha to the SO 2 moiety. This anion may then be quenched with a variety of electrophiles and subsequently deprotected to form the desired analogs of E2. Alternatively, this anion may be quenched with an aldehyde to form (after subsequent dehydration, i.e., an aldol-type condensation) an exo-methylene compound which may then be reduced hydrogenation) to form the desired analogs of E2. Analogously, preparation of E3 results from a Wittig reaction using methyl(triphenylposphoranylidene) acetate followed by simultaneous reduction of the double bond and cyclization using magnesium metal in methanol (Wei et al., Tetrahedron Lett., 34(28), pp. 4439-42 (1993)). A similar N-protection, deprotonation, quench and Ndeprotection scheme, or condensation-reduction scheme, as described in the preparation of E2, results in desired analogs of E3. Alternatively, E3 may be prepared from commercially available EVI. The hydroxyl group may be activated using commonly available reagents such as methanesulfonyl chloride or paratoluenesulfonyl chloride in the presence of a tertiary amine base. The addition of a nucleophile to displace the mesylate or tosylate yields EVII (Ackermann et al., Helv. Chim. Acta, 73, pp. 122-32 (1990)) which may be treated as described above to obtain E3.
Methods for the preparation of compounds E4- E6 are also well known in the art and stem from readily available protected amino aldehydes. Treatment of these aldehydes with a variety of amines under reductive amination conditions well known in the art, such as sodium cyanoborohydride using DMF/Acetic acid
I
WO 97/27180 PCT/US97/01610 112 as a solvent mixture, followed by deprotection of the primary amine yields diamine EVIII. Intramolecular cyclization with a variety of activated carbonyl, dicarbonyl or sulfuryl equivalents in the presence of a tertiary amine base yields compounds E4-E6. Examples of activating reagents include but are not limited to carbonyldiimidazole, phosgene, sulfuryldichloride, sulfuryldiimidazole, sulfonyl diimide, and oxalyl chloride.
Methods leading to the production of analogs of compound E7 are also known in the art (McManus et J. Med. Chem., 8, pp. 766-76 (1965)). Scheme 3 exemplifies several potential routes to the synthesis of compound E7. Any protected amino alcohol may be deprotonated to form the alkoxide which may be reacted with a substituted alpha bromo ester to form ether EIX (route Alternatively (route EIX may be formed from activation of a protected amino alcohol with, for example, methanesulfonyl chloride or paratoluenesulfonyl chloride in the presence on a tertiary amine base and subsequent addition of a nucleophile such as an alkoxide from an alpha hydroxy acid to displace mesylate or tosylate to yield EIX. Compound EIX can then be deprotected, free based with a tertiary amine base or potassium carbonate in methanol, and heated to effect cyclization to form E7. Alternatively (route E7 may be prepared from a protected amino alcohol by protection of the hydroxyl group with, for example, t-butyldimethyl silyl chloride/imidazole to afford the silyl ether. Subsequent nitrogen deprotection and acylation with a alpha bromo acid in the presence of any number of available coupling agents (for example dicylcohexylcarbodiimide, other related carbodiimide reagents or isobutyl chloroformate) or WO 97/27180 PCT/US97/01610 113 acylation with an alpha bromo acid chloride provides compound EX. Desilylation using, for example, tetrabutylammonium formate in THF followed by formation of the alkoxide with base affords cyclization to E7.
Alternatively, E7 may be prepared from the corresponding a-methylene compound both R 2 are H in E7, the nitrogen may be protected if necessary) by a multiple deprotonation-alkylation sequence to give an E7 wherein each R is inserted in an independent alkylation step and each R 2 may be attached to form a spirocyclic product alkylation with a dihaloalkane).
Schemes 4-6 describe methods for converting the cyclic compounds E1-E7 into compounds of this invention. For example, compounds of the type Z, exemplified by compounds E1-E7, may be deprotonated and reacted with a functionalized epoxide to generate the desired compounds as described in Scheme 4.
Several of the described epoxides are readily synthesized via methods well known in the art (Maligres et al,, Tetrahedron Lett., 36, pp. 2195-98 (1995)).
Optionally, further modification of the compounds may be performed subsequent to epoxide opening using reactions and materials well known in the art. For example, subsequent to epoxide opening utilizing example EXIb deprotection of the carbamate allows further modification of the unmasked amine.
Alternatively, as shown in Scheme compounds EZ may be converted to the desired products in a more stepwise fashion. Compounds EZ may be deprotonated using, for example, sodium hydride in DMF and treated with a three carbon based epoxide to generate epoxide EXII. Examples of such reagents include, but are not limited to, epibromohydrin, WO 97/27180 PCT/US97/01610 114 epichlorohydrin and glycidyl tosylate. Several other potential methods for preparing compounds of the type EXII are well known in the art, for example, the anion of Z may be reacted with allyl bromide or allyl iodide to form an allyl intermediate, which may subsequently be oxidized to form the desired epoxide. Several epoxidation conditions for the generation of either racemic or chiral epoxides are well known in the art.
Epoxide EXII may then treated with an amine and susequently carbonylated or sulfonated using activated species well known in the art to generate final -compounds of the type E9. Alternatively EXII may be reacted with a functionalized secondary amine followed by optional manipulation of R 2 to produce compounds of the type E10. One example of such manipulation is reaction of EXII with the known Boc piperazine EXIII (Dorsey et al., J. Med. Chem.,37, pp. 3443-51 (1994)).
Subsequent to epoxide opening, the Boc group may be removed and the unmasked secondary amine may be further manipulated by reaction with various electrophiles to form the desired product.
Scheme 6 describes a method for introduction of electrophiles into comounds of the type EXIV. Said compounds may be protected with a variety of protecting groups, for example t-butyldimethylsilyl triflate, to mask the secondary hydroxyl group followed by treatment with a non-nucleophilic base such as lithium diisopropylamide or hexamethyldisilyzane to generate the anion alpha to the carbonyl. Various electrophiles may then be added to substitute the position alpha to the carbonyl, or alternatively an aldol-type condensation-reduction scheme may be employed.
Deprotection of the secondary hydroxyl then yields the desired product.
WO 97/27180 PCTUS97/01610 115 As can be appreciated by the skilled artisan, the above synthetic schemes.. are not intended to comprise a comprehensive list of all means by which the compounds described and claimed in this application may be synthesized. Further methods will be evident to those of ordinary skill in the art.
Moreover, the determination of the optimum overall scheme, as well as the choice of reagents and reactions used to carry out the various steps in a given scheme will be based upon factors that are readily apparent to those of skill in the art. These factors include the identity of the compound to be produced, the efficiency of the individual steps and schemes in producing that compound in terms of overall yield, time, and cost and availability of reagents. It will therefore be apparent that some routine experimentation may be required in determining the optimum scheme to produce certain compounds of this invention.
It should be understood that the compounds of this invention may be modified by. appending appropriate functionalities to enhance selective biological properties. Such modifications are known in the art and include those which increase biological penetration into a given biological compartment blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism and alter rate of excretion.
The compounds of this invention are characterized by a superior ability to inhibit protease activity and viral replication, particularly aspartyl protease activity. These compounds are especially well suited for inhibiting HIV aspartyl protease. We WO 97/27180 PCT/US97/01610 116 believe that this activity is due to specific steric and electronic interactions between the protease and compounds of this invention. This belief stems from our analysis of the structural basis for the activity of compounds of this invention, in view of the known crystal structures of.HIV protease and bound inhibitors, such as the structure reported in Miller et al. "Structure of Complex of Synthetic HIV-1 Protease with a Substrate-Based Inhibitor at 2.3 A Resolution", Science, vol. 246, pp. 1149-1152 (1989), which is incorporated herein by reference, as well as structures ,determined in our laboratories.
The novel compounds of the present invention are excellent ligands for aspartyl proteases, particularly HIV-1 and HIV-2 proteases. Accordingly, these compounds are capable of targeting and inhibiting late stage events in HIV replication, the processing of the viral polyproteins by HIV encoded proteases. Such compounds inhibit the proteolytic processing of viral polyprotein precursors by inhibiting aspartyl protease. Because aspartyl protease is essential for the production of mature virions, inhibition of that processing effectively blocks the spread of virus by inhibiting the production of infectious virions, particularly from chronically infected cells. Compounds according to this invention advantageously inhibit the ability of the HIV-1 virus to infect immortalized human T cells over a period of days, as determined by an assay of extracellular p24 antigen a specific marker of viral replication.
Other anti-viral assays have confirmed the potency of these compounds.
The compounds of this invention may be employed in a conventional manner for the treatment of WO 97/27180 PCT/US97/01610 117 viruses, such as HIV and HTLV, which depend on aspartyl proteases for obligatory events in their life cycle.
Such methods of treatment, their dosage levels and requirements may be selected by those of ordinary skill in the art from available methods and techniques. For example, a compound of this invention may be combined with a pharmaceutically acceptable adjuvant for administration to a virally-infected patient in a pharmaceutically acceptable manner and in an amount effective to lessen the severity of the viral infection or to alleviate pathological effects associated with ,HIV infection or immunosuppression such as opportunistic infections or various cancers, tumors, CMV retinitis, candida infections, maternal fetal transmission, and AIDS related dementia,.
Alternatively, the compounds of this invention may be used in prophylactics and methods for protecting individuals against viral infection during a specific event, such as childbirth, or over an extended period of time. The compounds may be employed in such prophylactics either alone or together with other antiretroviral agents to enhance the efficacy of each agent. As such, the novel protease inhibitors of this invention can be administered as agents for treating or preventing HIV infection in a mammal.
The compounds of formula I, especially those having a molecular weight of less than about 700 g/mole, may be readily absorbed into the bloodstream of mammals upon oral administration. Compounds of formula I having a molecular weight of less than about 600 g/mole and aqueous solubility of greater than or equal to 0.1 mg/mL are most likely to demonstrate high and consistent oral availability. This surprisingly impressive oral availability makes such compounds WO 97/27180 PCT/US97/01610 118 excellent agents for orally-administered treatment and prevention regimens against HIV infection.
The compounds of this invention may be administered to a healthy or HIV-infected patient either as a single agent or in combination with other anti-viral agents which interfere with the replication cycle of HIV. By administering the compounds of this invention with other anti-viral agents which target different events in the viral life cycle and which target different viral substrains with varying susceptability to specific agents, the therapeutic effect of these compounds is potentiated. For instance, the co-administered anti-viral agent can be one which targets early events in the life cycle of the virus, such as cell entry, reverse transcription and viral DNA integration into cellular DNA. Anti-HIV agents targeting such early life cycle events include, didanosine (ddl), dideoxycytidine (ddC), d4T, zidovudine (AZT), 3TC, 935U83, 1592U89, 524W91, polysulfated polysaccharides, sT4 (soluble CD4), ganiclovir, trisodium phosphonoformate, eflornithine, ribavirin, acyclovir, alpha interferon and trimethotrexate. Additionally, non-nucleoside inhibitors of reverse transcriptase, delavirdine (U90) or nevirapine, may be used to potentiate the effect of the compounds of this invention, as may viral uncoating inhibitors, inhibitors of trans-activating proteins such as tat or rev, or inhibitors of the viral integrase.
Combination therapies according to this invention exert an additive or synergistic effect in inhibiting HIV replication because each component agent of the combination acts on a different site of HIV replication or on different strains of virus present in WO 97/27180 PCT/US97/01610 119 an infectious population. The use of such combination therapies may also advantageously reduce the dosage of a given conventional anti-retroviral agent which would be required for a desired therapeutic or prophylactic effect, as compared to when that agent is administered as a monotherapy. Such combinations may reduce or eliminate the side effects of conventional single antiretroviral agent therapies, while not interfering with the anti-retroviral activity of those agents. These combinations reduce potential of resistance to single agent therapies, while minimizing any associated toxicity.
Advantages of combining HIV protease inhibitors may include viral population effects, whereby certain members of a virus population which show reduced sensitivity to one protease inhibitor may be fully sensitive to another inhibitor or may in fact have enhanced sensitivity to the second inhibitor.
Alternatively or in addition, administration of two or more different inhibitors may be used to reduce specific toxicities associated with a single agent.
This advantage of combination therapy also applies to co-administration of the protease inhibitor of this invention with other antiviral agents. Alternatively or in addition, co-administration of more than one protease inhibitor may lower the rate of metabolic inactivation of the compounds of this invention, for instance, by inhibiting enzymatic systems such as cytochrome P 450 or esterases or the like. In particular, co-administration of compounds of this invention with protease inhibitors such as ritonavir or other agents such as ketoconazole, grapefruit juice and antiulcer medications such as H 2 -blockers, which WO 97/27180 PCT/US97/01610 120 inhibits cytochrome P 450 3A 4 may advantageously enhance their biological half-life.
These combinations may also increase the efficacy of the conventional agent without increasing the associated toxicity. Compounds of this invention in combination with other anti-HIV agents may act in an additive or synergistical manner in preventing the replication of HIV in human T cells. Preferred combination therapies include the administration of a compound of this invention with AZT, ddl, ddC, d4T, 3TC, 935U83, 1592U89, 524W91 or a combination thereof.
Alternatively, the compounds of this invention may also be co-administered with other HIV protease inhibitors such as VX-478 (Vertex, also known as 141W94 (Glaxo-Wellcome) and KVX-478 (Kissei)), saquinavir (Ro 31-8959, Roche), indinavir (L-735,524, Merck)), ritonavir (ABT 538, Abbott), nelfinavir (AG 1343, Agouron), palinavir (Bila 2011 BS), U-103017 (Upjohn), XM 412 (DuPont Merck), XM 450 (DuPont Merck), BMS 186318 (Bristol-Meyers Squibb), CPG 53,437 (Ciba Geigy), CPG 61,755 (Ciba Geigy), CPG 70,726 (Ciba Geigy), ABT 378 (Abbott), GS 3333 (Gilead Sciences), GS 3403 (Gilead Sciences), GS 4023 (Gilead Sciences), GS 4035 (Gilead Sciences), GS 4145 (Gilead Sciences), GS 4234 (Gilead Sciences), and GS 4263 (Gilead Sciences) or prodrugs of these or related compounds to increase the effect of therapy or prophylaxis against various viral mutants or members of HIV quasi species.
We prefer administering the compounds of this invention as single agents or in combination with retroviral reverse transcriptase inhibitors, such as nucleoside derivatives, or other HIV aspartyl protease inhibitors, including multiple combinations comprising from 3-5 agents. We believe that the co-administration WO 97/27180 PCT/US97/01610 121 of the compounds of this invention with retroviral reverse transcriptase inhibitors or HIV aspartyl protease inhibitors may exert a substantial additive or synergistic effect, thereby preventing, substantially reducing, or completely eliminating viral replication or infection or both, and symptoms associated therewith. Particularly preferred is administration of a combination of a compound of formula I, 3TC and zidovudine (AZT). Also preferred are administrations of combinations of a compound of formula I and 1592U89, or of compounds of formula I with VX-478, optionally with one or more reverse transcriptase inhibitors, paarticularly, AZT, 3TC and 1592U89.
The compounds of this invention can also be administered in combination with immunomodulators and immunostimulators bropirimine, anti-human alpha interferon antibody, IL-2, GM-CSF, interferon alpha, diethyldithiocarbamate, tumor necrosis factor, naltrexone, tuscarasol, and rEPO); and antibiotics pentamidine isethiorate) to prevent or combat infection and disease associated with HIV infections, such as AIDS, ARC and HIV-associated cancers.
When the compounds of this invention are administered in combination therapies with other agents, they may be administered sequentially or concurrently to the patient. The additional agents may be administered separately, as part of a multiple dose regimen, from the compounds of this invention.
Alternatively, those agents may be part of a single dosage form, mixed together with the compounds of this invention in a single composition. The pharmaceutical compositions according to this invention may comprise a combination of an aspartyl protease inhibitor of this WO 97/27180 PCT/US97/01610 122 invention and one or more therapeutic or prophylactic agents.
Although this invention focuses on the use of the compounds disclosed herein for preventing and treating HIV infection, the compounds of this invention can also be used as inhibitory agents for other viruses which depend on similar aspartyl proteases for obligatory events in their life cycle. These viruses include other AIDS-like diseases caused by retroviruses, such as simian immunodeficiency viruses, HTLV-I and HTLV-II. In-addition, the compounds of this invention may also be used to inhibit other aspartyl proteases, such as renin, pepsin, cymosin, RSV protease, AMV protease, SIV protease and FIV protease, and in particular, other human aspartyl proteases, including renin, and aspartyl proteases that process endothelin precursors.
Pharmaceutical compositions of this invention comprise any of the compounds of the present invention, and pharmaceutically acceptable salts thereof, with any pharmaceutically acceptable carrier, adjuvant or vehicle. Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) such as d-a-tocopherol polyethyleneglycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, polyethyleneglycol polymers such as PEG-400, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty WO 97/27180 PCT/US97/01610 123 acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat. Cyclodextrins such as and y-cyclodextrin, or chemically modified derivatives such as hydroxyalkylcyclodextrins, including 2- and 3-hydroxypropyl--cyclodextrins, or other solublized derivatives may also be advantageously used to enhance delivery of compounds of formula I.
The pharmaceutical compositions of this invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. We prefer oral administration or administration by injection. The pharmaceutical compositions of this invention may contain any conventional non-toxic pharmaceutically-acceptable carriers, adjuvants or vehicles. In some cases, the pH of the formulation may be adjusted with pharmaceutically acceptable acids, bases or buffers to enhance the stability of the formulated compound or its delivery form. The term parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or infusion techniques.
The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated WO 97/27180 PCT/US97/01610 124 according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterallyacceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are mannitol, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides.
Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant such as carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms such as emulsions and or suspensions. Other commonly used surfactants such as Tweens and Spans and/or other similar emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.
The pharmaceutical compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, hard or soft gelatin capsules, tablets, emulsions and aqueous suspensions, dispersions and solutions. In the WO 97/27180 PCT/US97/01610 125 case of tablets for oral use, carriers which are commonly used include lactose and corn starch.
Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions and/or emulsions are administered orally, the active ingredient may be suspended or dissolved in an oily phase combined with emulsifying and/or suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added.
The pharmaceutical compositions of this invention may also be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components. Such materials include, but are not limited to, cocoa butter, beeswax and polyethylene glycols.
Topical administration of the pharmaceutical compositions of this invention is especially useful when the desired treatment involves areas or organs readily accessible by topical application. For application topically to the skin, the pharmaceutical composition should be formulated with a suitable ointment containing the active components suspended or dissolved in a carrier with suitable emulsifying agents. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and water.
WO 97/27180 PCT/US97/01610 126 Alternatively, the pharmaceutical composition can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. The pharmaceutical compositions of this invention may also be topically applied to the lower intestinal tract by rectal suppository formulation or in a suitable enema formulation. Topically-transdermal patches are also included in this invention.
The pharmaceutical compositions of this invention may be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.
Dosage levels of between about 0.01 and about 100 mg/kg body weight per day, preferably between about and about 75 mg/kg body weight per day of the active ingredient compound are useful in the prevention and treatment of viral infection, including HIV infection. Typically, the pharmaceutical compositions of this invention will be administered from about 1 to about 5 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. A WO 97/27180 PCTUS97/01610 127 typical preparation will contain from about 5% to about active compound Preferably, such preparations contain from about 20% to about 80% active compound.
Upon improvement of a patient's condition, a maintenance dose of a compound, composition or combination of this invention may be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained when the symptoms have been -alleviated to the desired level, treatment should cease. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of disease symptoms.
As the skilled artisan will appreciate, lower or higher doses than those recited above may be required. Specific dosage and treatment regimens for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the infection, the patient's disposition to the infection and the judgment of the treating physician.
The compounds of this invention are also useful as commercial reagents which effectively bind to aspartyl proteases, particularly HIV aspartyl protease.
As commercial reagents, the compounds of this invention, and their derivatives, may be used to block proteolysis of a target peptide or may be derivatized to bind to a stable resin as a tethered substrate for WO 97/27180 PCT/US97/01610 128 affinity chromatography applications. For example, a compound of formula I may be tethered to an affinity column to purify recombinantly produced HIV protease.
Derivatization of the compounds of this invention to produce affinity chromatography resins and the methods used to purify proteases using such resins are well known and within the skill of the art. These and other uses which characterize commercial aspartyl protease inhibitors will be evident to those of ordinary skill in the art. (See: Rittenhouse, J. et al. Biochem.
Biophvs. Res. Commun. 171, p. 60 (1990) and Heimbach, J.C. et al. Ibid 164, p. 955 (1989)).
In order that this invention be more fully understood, the following examples are set forth.
These examples are for the purpose of illustration only and are not to be construed as limiting the scope of the invention in any. way.
General Materials and Methods All temperatures are recorded in degrees Celsius. Thin layer chromatography (TLC) was carried out using 0.25 mm thick E. Merck silica gel 60 F 254 plates and elution with the indicated solvent system.
Detection of the compounds was carried out by treating the plate with an appropriate visualizing agent, such as 10% solution of phosphomolybdic acid in ethanol or a 0.1% solution of ninhydrin in ethanol, followed by heating, and/or by exposure to UV light or iodine vapors when appropriate. Thick layer silica gel chromatography was also carried out using E. Merck
F
254 plates ("prep plates") of 0.5, 1.0, or 2.0 mm thickness. Following development of the plate, the WO 97/27180 PCT/US97/01610 129 band of silica containing the desired compound was isolated and eluted with an appropriate solvent.
Analytical HPLC was carried out using a Water's Delta Pak, 5 uM silica, C18 reversed-phase column, 3.9 mm ID x 15 cm L with a flow rate of 1.5 mL/min using the following table: Mobile phase: A 0.1% CF 3
CO
2 H in H 2 0 B 0.1% CF 3
CO
2 H in CH 3
CN
Gradient: T 0 min., A B T 20 min., A B (100%) T 22.5 min., A B (100%) Preparative HPLC was also carried out using C18 reversed-phase media. HPLC retention times were recorded in minutes. NMR spectral data was recorded using a Bruker AMX500, equipped with either a reverse or QNP probe, at 500 MHz, and was taken in the indicated solvent.
We have measured the inhibition constants of each compound against HIV-1 protease using the method described essentially by M.W. Pennington et al., Peptides 1990, Giralt, E. and D. Andreu, Eds., Escom, Leiden, Netherlands (1991); and the method described essentially by Partaledis et al., J. Virol., 69, pp.
5228-35 (1995).
Compounds of invention were tested for their antiviral potency in several virological assays. In the first assay, the compounds were added as a solution in dimethylsulfoxide (DMSO) to a test cell culture of CCRM-CEM cells, a strain of CD4+ human T-cell lymphoma cells, previously acutely infected with HIVIIIb using WO 97/27180 PCT/US97/01610 130 standard protocols (see Meek, T. D. et al., "Inhibition of HIV-1 protease in infected T-lymphocytes by synthetic peptide analogues", Nature, 343, p. (1990).
The effect of the compounds on inhibiting the replication of the virus was measured by determining the HIV extracellular p 24 antigen concentration using a commercial enzyme immunoassay (obtained from Coulter Corporation, Hialeah, FL).
Antiviral activity may also be measured in .a separate assay in MT4 cells. Antiviral HIV activity and compound-induced cytotoxicity.were measured in parallel by means of a propidium iodide based procedure in the human T-cell lymphotropicvirus transformed cell line MT4. Aliquots of the test compounds were serially diluted in medium (RPMI 1640, 10% fetal calf serum (FCS), and gentamycin) in 96-well plates (Costar 3598) using a Cetus Pro/Pette. Exponentially growing MT4 cells were harvested and centrifuged at 1000 rpm for minutes in a Jouan centrifuge (model CR 4 12). Cell pellets were resuspended in fresh medium (RPMI 1640, FCS, 20% IL-2, and gentamycin) to a density of 5 x 105 cells/ml. Cell aliquots were infected by the addition of HIV-1 (strain IIIB) diluted to give a viral multiplicity of infection of 100 x TCID50. A similar cell aliquot was diluted with medium to provide a mockinfected control. Cell infection was allowed to proceed for 1 hour at 37 °C in a tissue culture incubator with humidified 5% CO 2 atmosphere. After the 1 hour incubation the virus/cell suspensions were diluted 6-fold with fresh medium, and 125 ul of the cell suspension was added to each well of the plate WO 97/27180 PCT/US97/01610 131 containing prediluted compound. Plates were then placed in a tissue culture incubator with humidified CO2 for 5 days. At the end of the incubation period, 27 ul of 5% Nonidet-40 was added to each well of the incubation plate. After thorough mixing with a Costar multitip pipetter, 60 ul of the mixture was transferred to filter-bottomed 96-wellplates. The plates were analyzed in an automated assay instrument (Pandex Screen Machine, Baxter Biotechnology Systems). The assay makes use of a propidium iodide dye to estimate the DNA content of each well. The antiviral effect of a test compound is reported as an IC50, i.e. the inhibitory concentration that would produce a decrease in the HIV induced cytopathic effect. This effect is measured by the amount of test compound required to restore 50% of the cell growth of HIVinfected MT-4 cells compared to uninfected MT-4 cell controls.
References: 1. Averett, D.R. 1989. Anti-HIV compound assessment by two novel high capacity assays. J. Virol. Methods 23: 263-276.
2. Schwartz, et al. 1988. A rapid and simple colorimetric test for the study of anti-HIV agents.
AIDS Res. and Human Retroviruses, 4 441-447.
3. Daluge, et al. 1994. 5-chloro-2',3'dedeoxy-3'fluorouridine (935U83), a selective antihuman immunodeficiency virus agent with an improved WO 97/27180 PCT/US97/01610 132 metabolic and toxicological profile. Antimicro.
Aaents and Chemother., 38(7):1590-1603.
4. Dornsife, et al. 1991. Anti-human immunodeficiency virus synergism by zidovudine azidothymidine) and didanosine (dideoxyinosine) contrasts with their additive inhibition of normal human marrow progenitor cells. Antimicro. Agents and Chemother., 35(2): 322-328.
Depending on the cell type and the desired readout, syncytia formation, reverse-transcriptase (RT) activity, or cytopathic effect as assayed by a dye uptake method may also be used as readouts of antiviral activity. See H. Mitsuya and S. Broder, "Inhibition of the in vitro infectivity and cytopathic effect of human T-lymphotropic virus type III/lymphoadenopathyassociated virus (HTLV-III/LAV) by dideoxynucleosides", Proc. Natl. Acad. Sci. USA, vol. 83, pp. 1911-1915 (1986).
Insofar as the compounds of this invention are able to inhibit the replication of the HIV virus in
CD
4 cells of human lineage, they are of evident clinical utility for the treatment of HIV infection.
These tests are predictive of the compounds ability to inhibit HIV protease in vivo.
WO 97/27180 PCT/US97/01610 133 L~ I~~~YI I--Y Example 1 0 oxalyl chloride \N
DMSO
O NHBoc -78 0 C NHBoc OH~ -78 0 C
H
OH
H
N-(t-butoxycarbonyl)-L-phenylalaninol; 251.3 g/Mol 10.0g 39.8 mmol DMSO 78 g/Mol 3.80mL 49.0 mmol oxalyl chloride 126.9 g/Mol 3.82mL 43.8mmol triethylamine 101 g/Mol 23.0mL 160mmol methylene chloride 200 mL The oxalyl chloride was added dropwise to a solution of DMSO in methylene chloride at -78 OC.
After stirring for 10 minutes, the alcohol was added as a solution in methylene chloride. The reaction was then stirred at -78 °C for 45 minutes. At this time the triethylamine was added and a white precipitate formed. The reaction was then stirred 45 minutes at -78 °C and 45 minutes at 0 OC. The reaction was then quenched by the addition of a solution of 90g of citric acid in 300 mL of water. The organic portion of the reaction was then washed by (2 x 80 mL) of both saturated sodium bicarbonate and brine. The combined WO 97/27180 PCT/US97/01610 134 organic layers were then dried over sodium sulfate, filtered and concentrated in vacuo to leave a white solid. The aldehyde was then used without further purification in the reductive amination.
B.
0 'k-NHBoc
H
1 4 NH NaCNBH 4 HOAc/DMF H v-N ,NHBoc allyl amine aldehyde sodium cyanoborohydride
DMF
acetic acid (glacial) 57 g/Mol 6.0 62.8g/Mol 4.C mL 160 mmol est. 39.8 mmol )g 6.4 mmol 180 mL 1.8 mL The aldehyde of Example 1A was dissolved in 180 mL of DMF at 25 oC. This was followed by addition of the aldehyde and 1.8 mL of acetic acid respectively. After 2 hours sodium cyanoborohydride was added, as a solid.
The reaction was then stirred at 25 °C for 12 hours.
The reaction was then quenched by the addition of 50 mL of saturated sodium bicarbonate, and after 10 min.
diluted by 100 mL of diethyl ether. The organic portion was then washed by (2 x 50 mL) of both saturated sodium bicarbonate and brine. The combined organic layers were then dried over magnesium sulfate, filtered and concentrated in vacuo. The crude oil was WO 97/27180 PCT/US97/01610 135 purified by silica gel chromatography eluting with 30 ethyl acetate: hexane to provide 8.8 g of product (29.8 mmol,
C.
NH HBoc 1 1) HCI 2) TEA, CDI Boc amine HCl/dioxane deprotected diamine-2HCl carbonyl diimidazole triethylamine methylene chloride 291 g/Mol 4 N HC1 162.15g/Mol 6.8g 15 mL 3.83g 2.77g 12.7mL 550mL 23.4 mmol 14.7 mmol 17.1 mmol 179 mmol 0.03 M The Boc amine of Example 1B was stirred in 15 mL of 4N HC1 at 25 °C for 1.5 hours. The reaction mixture was then concentrated in vacuo to provide a white foaming solid. 3.83 mg of the deprotected diamine was dissolved in 500 mL of methylene chloride. To this, triethyl amine was added. After stirring for minutes, CDI was added (solid). The reaction was then stirred for 24 hours. This was followed by concentration in vacuo. The crude material was purified by silica gel chromatography, eluting with ethyl acetate, to provide 2.15 g (67 of the desired allyl urea.
WO 97/27180 WO 9727180PCTfUS97/01610 136 0hc N Ph 0 toluene:CH 2 0 P HH H 0 aldehyde methyl (triphenyiphosphoranyli dene) acetate toluene methylene chloride 1. 0 equiv., 1. 05 eq.
8 OmL 12 OmL Combine 7.9g of (S)-N-Boc-amino-3-phenyl-l-propanal, 4OmL of anhydrous toluene and 6OmL of anhydrous methylene chloride. Add 9.8g of the ylide followed by 2OmL of toluene and 6OmL of methylene chloride. Stir overnight at room temperature. After approximatly 18 hours the solvent was removed in vacuc and the residue was purified by flash chromatography (EtOAc/Hexane) to give 7.lg(77%) of the desired ester.
B.
0 Ph Ph0<N I- OMe
H
Mg (turnings) HCI MeOH Ph Y H WO 97/27180 PCT/US97/01610 137 1 ester 4.5g, 1.0 equiv.
2 magnesium turnings (Aldrich) 3.2g 10.0 eq..
3 2N HCl 10 eq.
To a solution of ester 1 in anhydrous methanol at 0 °C was added Mg turnings with stirring under N 2 Bubbling became evident within 1 hour. The reaction was then stirred at 0 °C for -2.5 hours then allowed to warm to RT overnight (TLC (95:5, CH 2 C12:MeOH) showed reaction complete. st. mat. Rf .84, prod. Rf The reaction was cooled to 0 OC, neutralized with 2 N HC1, diluted with water, and the volume reduced in vacuo.
The remaining aqueous layer was extracted with 3 portions of methylene chloride and the combined organic layers were washed with brine, dried (MgSO 4 filtered, and concentrated in vacuo.. The residue was then purified by silica get flash chromatography (CH 2 Cl 2 MeOH/CH 2 C12) to yield desired lactam product (1.74g,75% yield).
Literature reference: Tetrahedron. Lett., 1993, 34 pp. 4439-4442.
C.
Boc anhydride TEA, DMAP INH CH 2
CI
2
/CH
3 CN INBoc 0 0 WO 97/27180 PCT/US97/01610 138 1 lactam from 2B 1.0 equiv., 1.7g 2 BOC anhydride 2.5 equiv., 5.2g 3 triethylamine 2.0 equiv, 2.7mL 4 DMAP 1.2 equiv, 1.4g Lactam 1 was dissolved in methylene chloride (20mL) and to this solution was added a solution of Boc anhydride 2 in CH 2 C12 (10 ml)followed by triethylamine (2 eq) and DMAP (1.2 eq). After stirring for 4 hours at room temperature the reaction was refluxed for 4 hours and after this time, an additional 1.0g of Boc anhydride in acetonitrile (20mL) and 700uL of triethylamine were added. The reaction was stirred for 15 hours at room temperature. (TLC (95:5, CH 2 Cl 2 MeOH) Rf (st mat.) .31. Rf(prod) The solvent was then removed in vacuo and the residue was partitioned between methylene chloride and water. The organic layer was washed with water and brine, dried (MgSO 4 and filtered. The dried organic layer was then concentrated in vacuo and the residue was purified by silica gel chromatagraphy (CH 2 C12) to yield desired boc lactam 2 (2.3g, 86%).
D.
LDA
NBoc Allyl bromide STHF YNBoc
THF
0 0 WO 97/27180 PCT/US97/01610 139 1 BOC-lactam from 2C 1.0-equiv.,85 mg 2 Allyl Bromide, (Aldrich) 1.8 equiv. 51uL 3 LDA, 1.29M (Aldrich) 2.0 equiv 420 uL Boc-lactam 1 was dissolved in dry THF and cooled to -78 °C and to this solution was added LDA via syringe.
After stirring for 40 min. at -78 allyl bromide was added via syringe and the reaction was stirred for 3 hours after which time an additional amount of allyl bromide (17 ul) was added. The reaction was then stirred at -78 °C for 4 hours (TLC (5:95, MeOH:CH 2 Cl 2 Rf (st mat.) .34. Rf(2 diast.) .55 and.61). The reaction was then quenched with ImL saturated NaCl solution, and partitioned between saturated sodium bicarbonate and ethyl acetate. The organic layer was then washed with water and brine, dried (MgS04), filtered and concentrated in vacuo. The residue was purified by silica get chromatography to yield allylated product 2 (47mg, 48% yield).
E.
Ph Ph S 1) LDA, benzyl bromide 2)TFA O
O
1 2 A mixture of diisopropylamine (4.6 mL, 3 eq) and THF mL) was cooled to -78 OC, and to this solution was added n-butyl lithium (1.4 eq) via syringe. This WO 97/27180 PCT/US97/01610 140 mixture was warmed to -10 °C and stirred for 40 min, after which time the mixture was cooled back to -78 OC.
A solution of Boc lactam 1 (3.0 g, 1 eq) in THF (15 mL total) was added. The reaction mixture was then stirred at -78 oC for 40 min followed by the addition of benzyl bromide (1..45 mL, 1.1 eq) via syringe After stirring for 2.5 hours at -78 oC, the reaction was warmed to -45 °C and stirred an additional 1 hour. The reaction was then quenched at -78 oC, with 0.5 mL saturated NaCl solution. The reaction was warmed to room temperature, diluted with ethyl acetate and the organic layer was washed with water and satuated NaC1, dried (MgSO 4 and concentrated in vacuo. The residue was then dissolved in methylene chloride (50 mL) and to this solution was added triflouroacetic acid (8 mL, excess). After 4 hours the reaction was concentrated in vacuo, and partitioned between a saturated solution of sodium bicarbonate and ethyl acetate. The organic layer was washed with water and brine and then dried (MgSO 4 and concentrated in vacuo. The resulting residue was purified by flash silica get chromatography to give 726 mg of the desired benzyl lactam product 2 as a mixture of diastereomers.
Example 3
A.
2-bromo-propionyl bromide NaH O DIEA o O HO N HO N Br NH S THF 1 H o WO 97/27180 PCT/US97/01610 141 Synthesis of 2-oxo-3-methvl-6-phenvethlmethlmorpholin Dissolve S-(-)-2-Amino-3-phenyl-l-propanol (1.51 g, mmol) in THF (10 ml). To 0 °C solution add (rac)-2bromopropionyl bromide (1.04 ml, 10 mmol), followed by a dropwise addition of diisopropylethylamine (1.73 ml, mmol). Warm up to rt and continue stirring for min. Remove solvents in vacuo and remove salts by ethyl acetate/water extraction Following magnesium sulfate drying, the ethyl acetate layer is evaporated and residue redissolved in anhydrous THF.
To 0 OC solution of intermediate 2 add 13 mM of NaH (from 60% mineral oil dispersion, removed by washing, with hexane). Solution was warmed up to rt and reaction terminated (MeOH) after 1 hr. Residue left after solvents removal was again partitioned between ethyl acetate/water organic phases combined, dried with magnesium sulfate, filtered and evaporated, resulting in 1.20 g crude product. Silica gel chromatography (ethyl acetate) yielded 0.70 g of pure product, 34% yield. H NMR (CDC13): 7.25 6.75 (broad s, 1H), 4.19 1H, J=7.0 Hz), 3.76 (2H, d, J=7.5 Hz), 3.57 (1H, 2.90 (2H, 1.49+1.46 (both s, total integration 3H). CHN: 70.0 (calc: 70.2), 7.3 6.8 Mass Spec. (API-)=204 (M- Silica gel plates: Rf=0.19 (1/1 ethyl acetate/hexane). HPLC at 220 nm (YMC 0.46 cm x 25 cm C18 reverse phase) t=11.47 min (single peak), gradient: 0-100%B/30 min, 1.5 ml/min, A=O.I% TFA in water, B=0.1% TFA in acetonitrile.
WO 97/27180 PCT/US97/01610 142
B.
H 2-bromo-isobutyryl bromide NaH DIEA Na H o HO NW THF HO N Br
NH
HI
O
1 2 3 Synthesis of 2-oxo-3,3-dimethyl6phenvlmethvlmorpholine.
Dissolve 3.02g (20--mM) of S-(-)-2-Amino-3-phenyl-1-.
propanol in 10 ml THF. To 0 OC solution add 2- Bromoisobutyryl bromide (2.47 ml, 20 mmol), followed by dropwise addition of diisopropylethylamine (3.47 ml, mmol). Warm up to rt and continue stirring for min. Remove solvents in vacuo and remove salts by ethyl acetate/water extraction Following magnesium sulfate drying, the ethyl acetate layer is evaporated and residue redissolved in anhydrous THF.
Following silica gel chromatography (1/1 ethyl acetate/hexane), 1.20 g of intermediate 2 is isolated from mixture containing overacylation product.
To 0 oC solution of 2 in 4 ml of anhydrous DMF add 4 mM of NaH (from 60% mineral oil dispersion, removed by washing with hexane).
After 14 hrs at rt, the solvent was removed and solid residue partitioned between ethyl acetate/water (2X), organic phases combined, filtered, evaporated and (silica gel) chromatographed with ethyl acetate, WO 97/27180 PCT/US97/01610 143 resulting in 0.20 g of product homogenous by TLC, but heterogeneous by HPLC.
C.
Ph Ph Ph PPh h 0 1 0 NH" KM 7_ N, I NH 0 0 X=CI, I 1 2 3,4 5 6 Synthesis of 2-oxo-3.3-sDirocvclohexvl6phenvlmethylmorpholine via multiple deprotonationalkylation route.
A solution of 1 (5.73 g) was dissolved in 5 ml of anhydrous DMF, cooled down to 00 C and 0.72 g of NaH was added portionwise. After stirring for 15 min at room temperature, the solution was cooled to 0 0 C and 4.70 g of p-methoxy-benzyl chloride was added. The reaction was then stirred at room temperature for two hours, followed by silica gel purification, yielding 4.72 g of 2.
M =312.1 1H NMR (CDC13)=7.26-6.87 (9H,m), 5.42 3.85 4.34 4.20 (d,1H), 3.79 3.68 3.42 3.26 (1H,m), 2.95 (2H, m).
4.70 g of 2 was dissolved in 10 ml of anhydrous THF, cooled to -78 OC and 9.8 ml of 2M LDA in heptane/THF/ethylbenzene was added. After 15 min, 4.56g of 1-chloro-5-iodopentane was added dropwise and the WO 97/27180 PCT/US97/01610 144 reaction carried out at -78 OC for 1 hr and then quenched. The solvents were removed and the material was purified by silica gel (2.6g, The resulting compound was ca 1:1 mixture of two diastereomers.
MS (API+)=416.2 1H NMR (CDC13)= 7.4-6.9 (9H, m), 5.40 4.23 3.83 3.80 3.75 3.55 3.36 3.12 2.96 1.88 1.58 (m,4H).
2.6 g of 3 was dissolved in 5 ml of acetone. 1.87 g of sodium iodide was added and refluxed overnight. Acetone was then removed in vacuo and the crude material purified by ethyl acetate/aqueous extraction, resulting in 2.8g of 4 MS (API+)=508.1 530.1 (M+Na).
2.8g of 4 was dissolved in 40 ml of anhydrous THF, cooled down to -78 oC, and 3.6 ml of 2M LDA was added.
The reaction was allowed to progress for 2 hrs, with gradual temperature increase to room temperature. The residue was quenched with water, THF was evaporated and the crude material desalted between ethyl acetate/water, resulting in 1.90 g of 1H NMR (CDC13)=7.35-6.83 5.35 3.79 3.76 3.55 3.23 2.0-1.05 1.90g of 5 was deprotected by 9.61 g of CAN in 3/1 acetonitrile/water overnight at room temperature.
The product 6 (0.50g) was purified on silica using EtOAc/hexane/methanol gradient.
WO 97/27180 PCTIUS97/01610 145 M =259 1H NMR (CDC13)=7.22 5H), 6.96 3.82 3.67 3.60 2.83 2.0-1.20 Example 4
A.
1) Trimethylsilyl methylmagnesium bromide, THF 2) borontrifluoride etherate O NHCbz NHCbz
H
1 2 7.Og of the aldehyde 1 was dissolved in 40 mL of THF and added dropwise to a cooled (-780) solution of 128 mL (128mMol) of 1M trimethylsilyl methylmagnesium bromide in ether. The resulting mixture was allowed to warm to rt and poured into water. After diluting with ethyl acetate and 1N HC1, the layers were separated and the organic layer was washed with 10% aqueous sodium bicarbonate. Drying over magnesium sulfate and removal of the solvent in vacuo gave a viscous oil, which was re-dissolved in 150 mL of dichloromethane and treated dropwise with 15.6 mL of borontrifluoride etherate.
The resulting mixture was stirred for 5 days at rt and then quenched with 10% NaOH. The organic layer was dried and evaporated and the residue was chromatographed on silica gel (20% ethyl acetate/hexanes) to give 5.2g of a yellow solid.
Recrystallization from hexane yielded 4.6g of the desired alkene as a white solid in three crops.
WO 97/27180 PCTIUS97/01610 146 '-NHCbz 1 Thioacetic acid AIBN 0 2 NHC 2 (7.lmMol) of the alkene from the previous step were mixed with 10 mL of carbon tetrachloride and 1.4 mL (20mMol) of thioacetic acid. A spatula tip of AIBN was added and the mixture was irradiated in a quartz vessel at 254nm for 2h. The resulting mixture was diluted with dichloromethane and extracted with satd.
aqueous sodium bicarbonate. Drying and removal of the solvent, followed by chromatography on silica gel ethyl acetate/hexane) gave the desired thioacetate as a pale yellow liquid which solidified on standing.
C.
0 NHCbz C12 acetic acid HCI aq.
C102S NHCbz HBr/HOAc ,N
ONH
A solution of 0.85g of the thioacetate from the previous step in 30 mL of acetic acid and 15 mL of 1N HC1 was cooled on ice and exposed to a stream of chlorine gas for 2h. Ethyl acetate was added and the organic layer was separated, dried and co-evaporated WO 97/27180 PCT/US97/01610 147 with toluene to give the desired sulfonyl chloride as a white solid (1.05g).
0.7g of the sulfonyl chloride 2 obtained in the previous step were dissolved in 30 mL of 30% HBr in acetic acid. After 2h, the volatiles were removed in vacuo, the gummy residue was redissolved in 100mL of chloroform and the solution was treated with ImL of triethylamine. The mixture was stirred for lh and then extracted with lN HC1 and 10% aqueous sodium bicarbonate. Drying over magnesium sulfate and removal of the solvent gave a brown oil which was chromatographed on silica gel MeOH/dichloromethane) to give the desired sulfonamide as an off-white solid (0.305g). 1H-NMR (CDC13): 2.20 2.48 (1H,m), 2.89 3.10 3.23 1H,m), 3.84 (1H, m), 4.18 (1H, bs), 7.30 13C-NMR (CDC13): 28.8, 42.0, 47.8, 56.2, 127.8, 129.1, 129.3, 136.6.
Example Synthesis of Sulfamate
A.
1) methylamine, EDCI, DMF 2) H 2 O NHCbz O NH2 OH NHCH3 1 2 A solution of 30g of Cbz-(L)-phenylalanine, 6.8g of methylamine hydrochloride, 14.8g of hydroxybenzotriazole and 22 mL of N-methylmorpholine in WO 97/27180 PCT/US97/01610 148 300 mL of dimethylformamide was cooled on an ice-bath and treated with 19.2g of EDCI. The mixture was allowed to reach rt overnight and then poured into 2000 mL of water. The product was collected by filtration, dried and redissolved in 500mL of methanol and 300 mL of THF. Ig of 5% palladium on carbon was added and the mixture was stirred under hydrogen for 36h. Filtration and removal of the solvent, followed by short plug filtration through silica gel MeOH(2M NH3)/ dichloromethane) gave the desire amine as a pale yellow solid (17g).
B.
S1) LiBH 4 TMS-CI, THF 2) sulfonyldiimide, pyridine
NHCH
3 1 2 A solution of 1.22g (56 mMol) of lithiumborohydride in 28 mL of THF was treated with 14.2 mL (112mMol) of chlorotrimethyl silane. The resulting mixture was treated scoopwise with 5g (28mMol) of the amide from the previous step. After stirring at rt for 24h, 40 mL of methanol were added carefully, followed by 10 mL of acetic acid. Repeated evaporation from methanol gave a colorless glass, which was dissolved in 100mL of NaOH. Extraction with 4x50mL of chloroform, followed by drying and removal of the solvent gave a yellow oil which was chromatographed on silica gel methanol(2M ammonia)/dichloromethane to give 1.5g of WO 97/27180 PCT/US97/01610 149 the desired diamine as a colorless oil, and 2.0g of recovered starting material.
0.15g of the diamine from the previous step were dissolved in 0.5 mL of pyridine and added dropwise to a refluxing solution of 0.lg of sulfonyldiimide in of pyridine. Reflux was continued for 24h and the volatiles were removed in vacuo. The resulting brown oil was chromatographed on silica gel (20% methanol(2M ammonia)/dichloromethane) to give the desired sulfonylurea as a yellow oil (0.04g). 1 H-NMR (CD 3 0D): 2.60 2.86 (1H,dd), 2.96 (1H,dd), 3.15 (lH,dd), 3.47 (1H,dd), 4.18 (lH, 7.22 7.38 1C-NMR (CD 3 0D): 31.8, 39.9, 50.0, 57.8, 126.5, 128.2, 129.0, 136.6 Example 6 Ph LDA Ph Boc EtOTC CI. EtO N 0 1THF 2 Boc lactam 1 (1.27 g, leq) was dissolved in THF (27 mL) and cooled to -78 OC. To this solution was added LDA (Aldrich, 1.5 M in hexane, 3.7 mL, 1.2 eq) via syringe over 3 minutes. After stirring for 85 minutes at 78 OC, a solution of ethyl iodoacetate (600 uL, 1.1 eq) in THF (13 mL) was added via syringe over 6 minutes. The reaction was then stirred at -78 OC for hours, then at 1.5 hours at -40 OC. The reaction was then cooled back to -78 OC and quenched with 2.5 mL WO 97/27180 PCT/US97/01610 150 saturated NaC1 solution, and partitioned between saturated sodium bicarbonate and ethyl acetate. The organic layer was then washed with brine, dried (MgSO 4 filtered and concentrated in vacuo. The residue was purified.by flash silica get chromatography eluting with 5% EtOAc/ CH2C1 2 to give 1.67g of substituted lactam product 2 contaminated with a minor amount of lactam starting material 1. HPLC showed 52% product and 28% starting material. This mixture was then dissolved in methylene chloride (45 mL) and cooled to 0 OC. To this solution was added trifluoroacetic acid (2 mL) and the reaction was stirred at room temperature for 1.5 hr. TLC showed no BOC material and the reaction was concentrate in vacuo and partitioned between saturated bicarbonate solution and ethyl acetate. The organic was washed with water, brine and dried (MgSO 4 The organic layer was evaporated in vacuo, and the residue was purified by flash chromatography eluting with 3:1 EtOAc/ hexane to give 770mg of pure lactam product 2.
Example 7 O\ 1) TMSI
TMEDA
-150 C, O 2) -150 Cto o C, 3 0minI H 0 O 2 A solution of 5-benzyl-pyrrolidinone 1 (1.5 gr, 8.86 mmol) was dissolved at ambient temperature under WO 97/27180 PCT/US97/01610 151 nitrogen in anhydrous dichloromethane (40 mL). TMEDA mL, 42.8 mmol) was added via pipette and the solution was cooled and maintained at -20 OC. TMSI (2.33 mL, 17.12 mmol) was added via pipette and the mixture was stirred for 15 min. Solid iodine (4.345 g, 17.12 mmol) was added and the mixture was stirred vigorously for 15 minutes and then quenched by rapid addition of the reaction mixture into aqueous sodium sulfite solution (100 mL). The mixture was transferred to a separatory funnel and the layers were separated. The organic layer was washed with IN NaHSO 4 water, and then dried over MgSO 4 The solution was then diluted in half with methanol and stirred overnight under a nitrogen atmosphere. The solvent was removed in vacuo and the residue was purified by flash chromatography, eluting with ethyl acetate hexane Pure iodo lactam product 2 was recovered as a solid (2.11 g).
Example 8
A.
Bn I Bn Bn'N OH 1) Ms-Cl, TEA, CJCl 2 Bn-'-OMs 1G 2 To a solution of dibenzylphenylalinol 1 (100 mmol) in methylene chloride (100mL), was added triethylamine (150 mmol). The mixture was cooled to 0 °C and methanesulfonyl chloride (110 mmol) was slowly added.
The mixture was stirred at 0 °C for one hour and then WO 97/27180 PCT/US97/01610 152 poured into a beaker containing diethyl ether (400mL).
The mixture was filtered and washed with more diethyl ether and the filtrate was washed with water, saturated NaHCO 3 and saturated brine. The organic layer was then dried (MgSO 4 filtered and concentrated to yield 41 g of crude mesylate product 2 as a light yellow-brown thick oil, which was used as is in subsequent steps.
B.
Bn Bn Bn- OMs K2Diethymalonate N CO2Et
K
2 C0 3 0 CCN
CO
2 Et 0 C K.
1 2 Diethyl malonate (300 mmol) was dissolved in acetonitrile (250 mL) and to this solution was added potassium carbonate (300 mmol); the suspension was stirred overnight at room temperature. Mesylate 1 (100 mmol) in acetonitrile (60mL) was then added to the reaction mixture which was then heated to 80 OC and stirred overnight. The reaction mixture was then filtered and concentrated in vacuo. Addition of hexane to the residue formed a precipitate, which was filtered as pure malonate product 2 (19.5 Material was used as is.
WO 97/27180 PCT/US97/01610 153
C.
Bn 1) NaH, THF, O'C- RT n yCOEt -OTT 2 Bn
CO
2 Et S CO 2 Et T 01 73% CO2Et 3 Malonate 1 (10.6 mmol) was dissolved in dry THF (40 mL) and cooled to 0 OC. To this solution, sodium hydride (17 mmol) was added in portions and the suspension was stirred for 1.5 hr at 0 OC. The triflate 2 (12 mmol) in dry THF (10mL) was then slowly added to the reaction mixture and after complete addition the reaction was allowed to warm to room temperature and was stirred overnight. The reaction was then diluted with water (100mL) and extracted with diethyl ether (3x50 mL).
The combined organic layers were then washed with saturated brine, dried over MgSO 4 filtered and concentrated in vacuo. The crude product was purified by mplc (eluted with a gradient of 9:1 hexane:ethyl acetate up to 4:1 hexane:.ethyl acetate to yield product 3 (4.2 g, 73
D.
0 Bn B CO 2 Et 1) HCI/H 2 Pd/C Bn 2) CH 2
CI
2 sat. NaHCO3
O
2 Et 0 2 E t 90% yield
NH
1 2 The subsituted malonate 1 (1.62 mmol) was suspended in ethanol and to this was added conc. HC1 (0.24 mL, 2.4 WO 97/27180 PCT/US97/01610 154 mmol) and 10% palladium on Carbon (0.162 mmol). This mixture was then stirred under a balloon of hydrogen gas at room temperature overnight. The reaction was then filtered through Celite and to the filtrate was added triethylamine (10 mL, excess) followed by solid sodium bicarbonate (excess). The mixture was stirred for 0.5 hr, filtered and concentrated to yield a yellow solid. This residue was then dissolved in ethyl acetate and washed with water, 0.5N HC1, saturated sodium bicarbonate,-and brine. The organic layer was dried (MgSO 4 filtered, and dried to yield crude .lactam product 2, which was used as is.
E.
1) KOH, ethanol 2) TsOH, DMSO, 80*C
NH
Et 2 C NH
O
O
1 2 Lactam 1 (1.18 mmol) was dissolved in ethanol (5mL) and to this solution was added KOH (10 mmol). The mixture was stirred for 3 hr at room temperature and then concentrated to dryness. The residue was dissolved in water and washed with diethyl ether. The aqueous layer was then acidified with HC1 and extracted with ethyl acetate. The organic layer was dried (MgSO4), filtered and concentrated in vacuo to yield 341 mg of a light yellow solid. The residue was dissolved in DMSO (3mL) and to this solution was added p-toluenesulfonic acid mono- hydrate, and the mixture was heated to 80 °C WO 97/27180 PCT/US97/01610 155 overnight. The mixture was diluted with water (15 mL) and extracted with ethyl acetate. The organic layer was washed with saturated sodium carbonate and brine followed by drying with MgSO 4 The organic layer was then filtered and concentrated in vacuo to yield the THF substituted lactam product (245 mg, 77% from ester) which was used as is in the next step without further purification.
Example 9
-A.
-Ph NaH, Ph OCp-methoxybenzyl chloride NH DMF O 0 C ->RT O 0 1 2 Sodium hydride (60% dispersion in mineral oil, 4.0 g, 1.17 eq) was washed with 4 x 25 mL portions of hexanes to remove the mineral oil, then suspended in 25 mL of DMF and cooled to 0 A solution of lactam 1 (15g, 1 eq) in dry DMF (25 mL) was then added dropwise via canula into the cold NaH suspension over 40 min. An additional 65 mL of DMF was then added to aid stirring.
After stirring the anion for 1 hour, p-methoxybenzyl chloride (14.5 mL, 1.26 eq) was added over 5 min at 0 oC. The reaction was then allowed to warm to room temp. An additional amount of p-methoxybenzyl chloride was added to drive the reaction to completion. TLC (EtOAC) Rf lactam 1 0.21. Rf product 2 0.43.
WO 97/27180 PCT/US97/01610 156 After 3.5 hours, the reaction was poured into cold water and extracted twice with ethyl acetate. The combined organic layers were washed with water brine, dried (MgSO 4 and filtered. Concentration in vacuo afforded a crude solid which was purified by crystallization( 7:1.hexane:EtOAc) to yield the protected lactam product 2 (19g,
B.
TMSI
TMEDA
OMe 12 H, OMe
CH
2
CI
2
I
O'C, 1 2 To protected lactam 1 (328 mg, 1.11 mmol) and N,N,N ,N -tetramethylethylenediamine (Aldrich, equiv., 5.55 mmol, 645 mg, 838 ml) in 15 ml dichloromethane at -15 was added iodotrimethylsilane (Aldrich, 1.0 equiv., 1.11 mmol, 222 mg, 158 ml). After 15 min, iodine (Aldrich, 1.2 equiv., 1.33 mmol, 338 mg) was added in one portion and the reaction warmed to 0 oC. After 30 min the reaction was quenched with 5 ml each of 10% aqueous sodium sulfite and saturated aqueous sodium chloride. The orgnic layer was separated, dried over magnesium sulfate, filtered and concentrated in vacuo.
Purification by flash column chromatography (silica gel, 2.5 x 10 cm, 2.5% diethylether in dichloromethane) WO 97/27180 PCT/US97/01610 157 yielded 322 mg of diastereomeric iodolactam 2 as a white solid.
C.
o o Bu 3 SnH, SAIBN OM e S 1 I PhMe 11 OC, 16h 0 0 1 2 To iodolactam 1 (1.18g, 2.91 mmol) and methyl vinyl sulfone (Aldrich, 6.0 equiv., 17 mmol, 1.82 g, 1.5 ml) in 25 ml. refluxing toluene was added tributyltin hydride (Aldrich, 1.3 equiv., 3.79 mmol, 1.10 g, ml) and AIBN (Pfaltz Bauer, 0.12 equiv., 0.35 mmol, 57 mg) as a solution in 5 ml toluene over 1.2 h. After 16 h the solvent was removed in vacuo, and the residue taken up in 200 ml diethyl ether and stirred with 20 ml aqueous potassium fluoride (wt/v) at ambient temperature. After 3 h the orgnic layer was separated, dried over magnesium sulfate, filtered and concentrated in vacuo Purification by flash column chromatography (silica gel, 5 x 20 cm, 2:1 ethyl acetate/hexanes) yielded 0.31g of diastereomeric sulfone 2 as a white solid.
WO 97/27180 PCT/US97/01610 158 Example
A.
H
o NHCbz "NaCNBH 3 HCI p q 1% acetic acid/DMF HN NHCz 2 -To a solution of solution of Cbz-L-phenylalinal (13 g, 45.9 mmol) in 1%AcOH/DMF (200 mL) mL was added aminoisobutyic acid methyl ester hydrochloride 1 g, 55.1 mmol) with stirring at room temperature. Once homogeneous, solid sodium cyanoborohydride (8.6 g, 137.6 mmol) was added in one portion. Some bubbling was evident and the reaction was stirred overnight at room temperature. The reaction was quenched with water mL) and concentrated in vacuo to about 100 mL. The concentrate was diluted with ethyl acetate and washed with water and brine followed by drying (MgSO 4 The organic layer was evaporated in vacuo to yield a yellow residue which was purified by MPLC (elutant 1:2 ethyl acetate hexane) to afford amine product 2 (11.6 g, 66%).
WO 97/27180 PCT/US97/01610 159
B.
1) 30% HBr/HOAc HN HN NHC t z 2) DIEA/methanol
CO
2
CH
3 H 0 1 2 To a solution of amine 1 (1.41 gr, 3.7 mmol) in methylene chloride (25 mL) was added 30% HBr in acetic "acid (6 mL) via pipet. Vigorous gas evolution occurred and the reaction was allowed to stir overnight at room temperature. The mixture was then evaporated in vacuo and dried under high vacuum. The residue was then dissolved in methanol (25 mL) and to this solution was added diisopropylethylamine (5eq) and the reaction was stirred at room temperature overnight. The solvent was removed in vacuo and the residue was taken up in ethyl acetate and washed with water, saturated NaHCO 3 and brine. The organic layer was dried (MgSO 4 filtered and concentrated in vacuo to yield crude product Flash silica gel chromatography methanol methylene chloride) afforded pure piperazinone product 2 (556 mg, 70 WO 97/27180 PCT/US97/01610 160
C.
Benzyl bromide K2C03 NH Acetonitrile NH 0 O 1 2 To a solution of piperazinone 1 (556 mg, 2.55 mmol) and potassium carbonate (1.06 g, 7.6 mmol) in acetonitrile was added benzyl bromide (364 uL, 3 mmol) and the reaction was stirred at room temperature overnight.
The reaction was then filtered and concentrated in vacuo. The residue was dissolved in ethyl acetate, washed with water and brine and dried (MgSO 4 The organic layer was then removed in vacuo and the residue was flash chromatographed methanol in methylene chloride) to yield pure benzyl protected piperazinone product 2 (589 mg, Example 11
A.
1) Benzylamine O Ph EDCI, HOBT, Ph
NMM,DMF
2) 30% HBr/HOAc N-Cbz 3) Borane-THF
N
A solution of Cbz-(1)-Phenylalanine (15 gr, 50 mmol), HOBT (7.4g, 50mmol), N-methyl morpholine (5.5 mL, mmol) and benzylamine (6 mL, 55 mmol) in 250 mL of DMF WO 97/27180 PCT/US97/01610 161 was cooled to 0 OC and treated with EDCI (9.6 g, mmol). The resulting mixture was stirred at 25 OC for 12h and the volatiles were removed in vacuo.
Partitioning between ethyl acetate and IN hydrochloric acid, followed by extraction with 10% sodium bicarbonate, drying over magnesium sulfate and evaporation of the solvent afforded the desired amide as a white solid (19.5g).
19g of the above material were dissolved in 280mL of 30% hydrogen bromide in acetic acid and stirred at °C for 3h. The volatiles were removed and the 'residue was partitioned between water and ether. The aqueous layer was treated with excess 6N sodium hydroxide and extracted twice with ethyl acetate.
Drying over magnesium sulfate and evaporation of the solvent afforded the desired amine as a pale yellow oil (14.0g), which was redissloved in 200 mL of tetrahydrofuran and treated with 200 mL of 1M borane- THF in tetrahydrofuran. The mixture was stirred at 25 °C for 72h and then heated to reflux for 4h. The solution was cooled and treated with 100 mL of methanol under vigorous gas evolution. The volatiles were removed and the resulting residue was dissolved in 150 mL of concentrated hydrochloric acid. After refluxing for Ih, the volatiles were removed and the residue was dissolved in 300 mL of 3N sodium hydroxide. Extraction with 3 times 250 mL of dichloromethane, drying over magnesium sulfate and chromatography on 2 inches of silica gel methanol-dichloromethane) gave the desired diamine as a pale yellow honey (9.2g).
WO 97/27180 WO 9727180PCTJUS97/01610 -162
B.
Ph sulfonyldilmide
DMAP
N pyridine H N. KsS /NH 2 A solution of sulfonyldiimide (3.6 g, 36 xnmol) in 100 niL of pyridine was heated to refJ.ux and treated dropwise with a solution of the diamine 1 (7.2 g, rrmol) from the previous step in 20 niL of pyridine.
After 2h of reflux, 15 mL of triethylamine and 0.4g of 4-dimethylaminopyridine were added and heating was continued for 12h. The volatiles were evaporated and the- residue was partitioned between 1N hydrochloric acid and ethyl acetate. Extraction of the organic layer with saturated sodium bicarbonate, drying over magnesium sulfate and chromatography on silica gel (1:1 ethylacetate hexanes) afforded the desired cyclic sulfamate 2 as a white solid 1H-FH4R (CDC13): 2.80(1H,dd), 2.96(1H,dd), 2.98(1H,dd), 3.32(lH,dd), 3.95(lH,m), 4.04(1H,d), 4.24(lH-,d), 4.40(1H,d) 7.18(2H,d) 7.2-7.4 (8H) 13C-NM'R(CDC13): 41.5, 50.0, 52.7, 53.8, 127.5, 128.0, 128.2, 128.3, 28.4, 128.5, 135.5, 136.0 WO 97/27180 PCT/US97/01610 163 Example 12
A.
Ph Ph II BnNH 2 OMs NH-Cbz SNal NH NH-Cbz 2 The Cbz-phenylalaninol mesylate 1 (280 mg, 0.77 mmol) was stirred in acetonitrile (5 mL) containing benzyl amine (413 mg, 3.85 mmol) and sodium iodide (115 mg, 0.77 mmol). The reaction was then refluxed for 24 hours. The reaction was then cooled to 25 °C and concentrated in vacuo. The crude oil was then purified by silica gel chromatography, eluting with CH 2 C12 with a gradient up to 1:1 CH 2
CI
2 :EtOAc to provide 120 mg of the desired diamine 2.
B.
Ph Ph 1 HBr NHCz HOAc NH2 N Cbz 1 2 The Cbz protected diamine 1 (120 mg, 0.32 mmol) was stirred in 2.0 mL of 30 HBr in acetic acid for one hour. This was followed by concentration in vacuo.
The crude oil was then dissolved into toluene and concentrated in vacuo two times followed by evacuation at approx. 1 mm Hg. The crude diamine was then purified by silica gel chromatography, eluting with WO 97/27180 PCT/US97/01610 164 95:5:1, CH 2 C1 2 :MeOH:NH 4 0H to provide 71 mg 90 of the desired diamine 2.
C.
Ph j Ph ^s.^NH I -I 1 2 0 The diamine 1 (56 mg, 0.23 mmol) was dissolved in 3.0 mL of CH 2 C1 2 This was followed by the addition of TEA (66 uL, 0.25 mmol) and then CDI (32 mg, 0.25 mmol).
A new spot was observed by tlc after 2-3 hours (Rf 0.29 in EtOAc on SiO 2 The reaction mixture was then concentrated and the residue was purified by silica gel chromatography, eluting with EtOAc, to provide 32 mg of the desired benzyl urea 2.
WO 97/27180 PCT/US97/01610 165 Example 13 Synthesis of Compound 1 DMF NN N NH OMO N NaH 0 0 OC ->RT 1 2 3 OMe allyl urea 216 g/Mol 100mg 0.46 mmol NaH, (60% in oil) 24 g/Mol 140.0 mg 9.7 mmol epoxide 325.4 g/Mol 150.0 mg 0.46 mmol DMF 2.0 mL The urea of Example 1C was dissolved in 1.0 mL of anhydrous DMF and cooled to 0 This was followed by the addition of 140 mg NaH. The reaction turned darker over the next hour at 0 oC. This was followed by the dropwise addition of the epoxide as a solution in DMF (0.6 mL), washing with 300 uL of DMF. The reaction was then stirred one hour at 0 followed by warming to Tlc indicated nearly complete conversion to two new products (Rf 0.4 and 0.45 on SiO 2 with 2:1 hexane: ethyl acetate, between that of the epoxide and the urea). The reaction was then cooled to 25 °C and quenched by the addition of 3 mL of saturated sodium r bicarbonate. The reaction mixture was then diluted by 15 mL of methylene chloride and washed by both saturated sodium bicarbonate and brine, (2 x 15 mL each). The organic portions were then dried over WO 97/27180 PCT/US97/01610 166 sodium sulfate, filtered and concentrated in vacuo.
The crude product was then purified by silica gel chromatography, eluting with 80% ethyl acetate: hexane to provide 35.0 mg of the desired alcohol.
Example 14
A.
DMF
OHa NH O NN->RT N ^NSO 2 2 3 OMe 1 lactam 1.0 equiv., 295mgg 2 sulfonamide epoxide 1.1 equiv., 520mg 3 NaH, 60% in oil (Aldrich) 1.5 equiv, 102mg 4 DMF 8 mL Lactam 1 was dissolved in 3mL of DMF and cooled to 00 C. To this solution was then added sodium hydride as a solid and the reaction was stirred for 40 min. at 0 The anion solution was canulated into a solution of epoxide 2 in 3 mL of DMF. The reaction was stirred at 0 °C for 5 minutes, then warm to room temperature and stirred overnight (TLC (95:5, CH 2 C12 MeOH) Rf (st mat.) .26. Rf(prod) After 22 hours, the reaction was cooled to 0 and quenched with
H
2 0/EtOAc. The organic layer was washed with and brine, dried (MgSO 4 filtered, and concentrated in WO 97/27180 PCT/US97/01610 167 vacuo. The residue was then purified by silica gel chromatography (40% ether/ CH 2 C12) to yield product 3 (310mg,37%).
Example
A.
OH P 0 2 0 TMS-triflate imidazole
DMF
-Sio 2 OMe 1 lactam t-butyldimethylsilyl trifluoromethanesulfonate imidazole 1.15g, 1.0 equiv.
1.5 equiv. .5 eq., (1.06mL) 2.5 equiv .5 eq, (470mg) Lactam 1 was dissolved in 5mL of DMF and cooled to 0 oC. To this solution was then added imidazole followed by TBDMS-triflate. The reaction was then allowed to warm to room temperature. After approximatly 2 hours, an additional .5 eq.(80mg) of TBDMS-triflate and .5 eq.(265uL) of imidazole was added and the reaction was stirred overnight. The reaction was quenched with saturated NaHCO 3 solution and partitioned between H 2 0/EtOAc. The organic layer was washed with water(5X) and brine, dried (MgSO 4 WO 97/27180 PCT/US97/01610 168 filtered, and concentrated in vacuo to yield product 2 gr,37%) which was used as is.
Example 16 Synthesis of Compound 7 -Si- 1) LDA ~Allyl bromide 6 P TBAF OMe 1 silyl-lactam 1.
Allyl Bromide, (Aldrich) 2.
LDA, 1.29M (Aldrich) 1.
TBAF, 1.OM, (Aldrich) 2.
OH
0 equiv.,23mg 1 equiv. 7 uL 25 equiv 36uL 5 equiv., Silyl protected lactam 1 was dissolved in THF and cooled to -78 To this solution, was added LDA (1.25 eq) via syringe. After stirring for 30 minutes at -78 allyl bromide was added via syringe. After 2 hours an additional 2ul of allyl bromide was added and the reaction was stirred at -78 °C for 2.5 hours, then warmed to room temp for 17 hours (TLC (2:8, ether:CH 2 C1 2 Rf (st mat.) .56. Rf(silyl-prod) After this time, TBAF (1M in THF) was added and the reaction was stirred at room temperature for 7 hours (TLC ether:CH 2 Cl 2 Rf(prod) The reaction mixture was then partitioned between H 2 0/EtOAc WO 97/27180 PCT/US97/01610 169 and the organic layer was washed with water and brine, dried (MgS04) and filtered concentrated in vacuo. The residue was then purified by silica gel chromatography ether/methylene chloride) to yield product 2 (6mg,30% yield).
Example 17 Synthesis of Compound silyl-lactam 1) LDA1.0 equiv., 122mg Sb nzBenzyl bromide 1. 2) TBAF2. OHe, O o 1 O 2 OMe OMe 1 silyl-lactam 1.0 equiv., 122mg benzyl bromide, (Aldrich) 1.5 equiv. 42uL LDA, 1.29M (Aldrich) 1.4 equiv 275u TBAF, 1.OM, (Aldrich) 2.5 equiv., 625uL Silyl lactam 1 was dissolved in dry THF (6mL) and cooled to -78 To this solution was then added LDA and the reaction was stirred for 30 minutes at -78 °C after which time benzyl bromide was added via syringe.
The reaction was stirred at -780C until reaction was complete (1.5 hours, TLC ether:CH 2 Cl 2 Rf (st mat.) .29. Rf(silyl-prod) .62. Rf(BzBr) .79).
The reaction was then quenched at -78 °C with 6uL water and then TBAF (1M in THF was added and the reaction was warmed to room temperature and stirred for 3 hours (TLC WO 97/27180 PCT/US97/01610 170 ether:CH 2 Cl 2 Rf(prod) The reaction was partition betweem H 2 0/EtOAc and the organic layer was washed with with water and brine, dried (MgS0 4 and filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (10% ether/
CH
2 C12) to yield benzyl product 2 (71mg, 48% Example 18 Synthesis of Compound 16 Methyl iodide F" O 2) TBAF OH 0 S0 02 O 1 2 O M e e 1 silyl-lactam 1.0 equiv.,66mg Methyl iodide, (Aldrich) 1.6 equiv., 16uL LDA, 1.29M (Aldrich) 1.3 equiv 1lOuL TBAF, 1.OM, (Aldrich) 3.0 equiv., 325uL: The reaction for the above methylated compound was carried out as per the procedure described for compound (Example 17) substituting methyl iodide for benzyl bromide on the scale described in the above table. The final compound was purified by silica gel chromatography using 10% ether/ CH 2 C1 2 to yield methylated product 2 (33mg, 60% yield).
WO 97/27180 PCT/US97/01610 171 Example 19
A.
I epibromohydrin,\
DMF
c e0 OC ->RT N 0 0 0 1 2 1 lactam 1.0 equiv., 400mg epibromohydrin 1.5 equiv., 280uL sodium hydride, 80% oil disp. 2.0 equiv, 126mg DMF Lactam 1 was dissolved in dry DMF (15 mL) and cooled to 0 °C under a nitrogen atmosphere. To this solution was added sodium hydride (2 eq) in one portion and the reaction was stirred at 0 °C for 1 hour after which, epibromohydrin was added via syringe. After stirring for 5 min. at 0 °C the reaction was warmed to room temperature (TLC (EtOAc) Rf (st mat.) .16.
Rf(prod) After 1.5 hours at room temperature the reaction was quenched with saturated NH 4 Cl and extracted with CH 2 C1 2 The organic layer was then washed with water(4X) and brine, dried (MgSO4) and filtered, and concentrate in vacuo. The residue was then purified by silica gel chromatagraphy (3:1 EtOAC:hexane) to yield 315mg(60%) of epoxide product 2 which was used as is in the next step.
WO 97/27180 WO 9727180PCTIJS97/01610 172 0 cyclopentylmethylamine, EtOH OH (OD 0 1 lactam.
cyclopentylmethylamine anhy. EtOH- 1.0 equiv., 315mg 5.75 equiv., 775mg 3mL Epoxide 1 was dissolved in 3 mL of EtOH and to this solution was added cylcopentylmethylamine. The reaction was heated to 80 0 C for 2.5 hours (TLC (9:1,CH 2 Cl 2 :MeOH) Rf (st mat.) .56. Rf(prod) The solvent was removed in vacuo and the residue was purified by silica gel chromatagraphy (3%MeOH/ CH 2 Cl 2 to l0%MeOH/ CH 2 Cl 2 to yield 224mg(50%) of amine product 2.
C. Synthesis of Comoound 1 )TM s-cl, TEAq lq 2) p-methoxybenzenesulfonyl chloride
O
4 hl ,oi-l P 3) TBAF NH ~S02 0 101 2 Ove WO 97/27180 PCT/US97/01610 173 1 lactam 1.0 equiv., 315mg chlorotrimethylsilane 2.2 equiv., 112uL triethylamine 5.0 equiv., 280uL 4-methoxybenzenesulfonylchloride 1.5 equiv., 124 mg TBAF, 1.OM 4.4 equiv., 1.78mL Amine 1 from Example 19B was dissolved in methylene chloride and cooled to 0 OC. To this solution was added triethylamine (2.5 eq) followed by chlorotrimethylsilane. The reaction was then warmed to room temperature and stirred under nitrogen for -hours. An additional amount of triethylamine was added eq) and 4-methoxybenzenesulfonyl chloride was added. The reaction was stirred at room temperature for 3 hours. After this time, TBAF (1M in THF) was added and the reaction stirred at room temperature for 1 hour. The solvent was removed in vacuo. and the residue partitioned between ethyl acetate and aqueous saturated bicarbonate solution. The organic layer was washed with water, brine, dried MgSO 4 filtered and the solvent removed in vacuo. (TLC CH 2 C12: ether), Rf(upper diast.) .21 Rf(lower diast.) .12).
The residue was purified by silica gel chromatagraphy ether/CH 2 C1 2 to yield 52 mg(26%) of (upper diastereomer). The lower diastereomer was further purified by preperative TLC ether:CH 2 Cl 2 to give 23mg(12%) of the lower diastereomer.
WO 97/27180 WO 9727180PCTIUS97/01610 174 Examrle Synthesis of Comroungi 47 P" DMF 0~ CH 2 m0 adtecolddwtoC befre adig020 soolnn wa remosoved in au and purfiendosc gel yielding 111 mg of final product 2 (compound 47) M =585 607.1 (M+Na) 1H NM~R (CDC13)= 7.52 (di, 2H), 7.30 (in, 5H), 6.95 (di, 2H), 4.05 (mn, 1H), 3.87 (3H, 3.60 (mn, 2H), 3.16 (mn, 4H), 3.0 (in, 4H), 2.13 (1H, mn), 1.97 (mn, 2H), 1.60 14H), 1.23 (mn, 4H).
WO 97/27180 PCT/US97/01610 175 Example 21 Synthesis of Compound 109 q NaH, DMF, 80*C 7 Sbromomethyl acrylic acid O N2) Ozone yOH 1 3) Dimethyl sulfide 2 0 thioproline-t-butyl amide, S' O EDC, HOBT. NMM, DMF I 2 H 0 N To a cooled solution (-78 OC) of benzyl lactam 1 (0.150g, 0.57 mmol) and bromomethyl acrylic acid (0.094g, 0.57mmol) in anhydrous THF (4.0mL) was added NaH 0.046g, 1.14 mmol) with stirring. The solution was allowed to gradually warm to room temperature and stir for 1.5h. The reaction mixture was then diluted with ethyl acetate (60mL) and washed with 1.ON HC1 (2 x 10mL) and brine (2 xlOmL). The organic layer was dried (magnesium sulfate), filtered, and evaporated to give an off white solid. This solid was dissolved in methylene chloride/methanol (80/20, and through the cooled solution(-78 oC) was bubbled ozone for 10min. The solution was flushed with oxygen, warmed to 0 oC, and methyl sulfide (2.0mL) was added at 0 oC. The mixture was allowed to warm to room temperature and stand for 1.0h. Evaporation of the solvent afforded crude product 2 as a yellow oil. To a solution of the acid 2 in anhydrous DMF (3.0mL) was added thioproline-t-butylamide (0.llg, 0.57mmol), hydroxybenzotriazole (0.77g, 0.57 mmol), N- methyl- WO 97/27180 PCT/US97/01610 176 morpholine (0.62mL, 0.57mmol) and EDCI (O.llg, 0.57 mmol) respectively with stirring at room temperature.
After 24h. at room temperature, the reaction mixture was evaporated and the residue was dissolved in ethyl acetate (100mL). The solution was washed with 1.0N HC1 (2 x 20mL), 10% sodium carbonate (2 x 20mL), water (1 x brine( 1 x 10mL), filtered and evaporated to give 0.210g of a yellow oil. The oil was purified by column chromatography; hexane/ethyl acetate (60/40) to give compound (0.050g, 18%) MS: M+l= 522; H NMR (chloroform-d) 1.35(d, 9H); 1.85(m,2H); 2.6(m, 3H); 2.85(m,1H); 3.15(m,2H); 3.40(m,1H); 3.8(m,1H); 4.1(m, 2H); 4.4(m,1H); 4.70(m,1H); 4.95(m, 1H); 6.1(d, 1H); 7.1(m,4H); 7.25(m,6H).
-Example 22 Synthesis of Compound S 1)NaH,DMF, 80°C H H Oconc. HCI O H H OH NH N 0 0 0 2 0.80g of allyl lactam 1 was dissolved in 1 ml of DMF, cooled to 0 °C and 89.5 mg of sodium hydride was then added. The solution was then brought up to ambient temperature for 30 min, again cooled down to 0 OC and 1.4 g of epoxide 2 was added. The reaction was warmed to 50 °C under N 2 blanket for 3 hrs. The resulting crude mixture was then chromatographed on silica gel WO 97/27180 PCT/US97/01610 177 yielding 1.4g of 3 This amount was treated with 12 ml of 4N HCl in dioxane and 2 ml water for min. The product was then chromatographed on C18rphplc, yielding 0.36g of two diastereomers, subjected to chiral separation, which resulted in 138 mg of pure diastereomer 3. MS (ES- 551.3 ES+, 553.3 (M+1) and 575.3 1H NMR (CDC13)= 7.20 14H), 6.26 1H), 5.62 1H), 5.24 1H), 4.97 2H), 4.23 1H), 3.83 2H), 3.61 1H), 2.95 2.40 1H), 2.24 1H), 2.04 1H), 1.95 2H), 1.70 2H).
Example 23 Synthesis of Compound 91 K 1) NaH, DMF, 80C S H conc. HCI OH(
OH
-,NNH N S3 2 a A solution of cyclic sulfamate 1 (0.1g, 0.33mmol) in 2 mL of dimethyl formamide was cooled to 0 °C and treated with of 60% sodium hydride (0.005g, 0.13 mmol) in oil.
The mixture was stirred at 25 °C for 1.5h and treated with of epoxide 2 (0.125g, 0.33mmol) The resulting mixture was stirred at 60 °C for 3h, more sodium hydride (0.005g) was added and heating was continued over night. The volatiles were removed in vacuo and the residue was dissolved in 2 mL of 4M hydrogen WO 97/27180 PCT/US97/01610 178 chloride in 1,4-dioxane. Water (0.5 mL) was added and the mixture was stirred for 6h at 25 The reaction mixture was diluted with ethyl acetate and extracted with 10% soduim bicarbonate. Drying over magnesium sulfate and removal of the solvents gave a yellow gum, which was subjected to C-18 preparative HPLC (acetonitrile-water gradient). The desired material 3 -was isolated as a minor fraction (9 mg) as a white solid 1H-NMR(CDC13): 2.10(2H), 2.70(2H), 2.8-3.2(8H), 3.4(1H), 3.58(1H), 4.02(1H), 4.15(1H), 4.22(2H), 5.30(1H), 5.86(1H), 7.06(2H), 7.1-7.4(16H).
Example 24 Synthesis of Compound 83 iq 1) NaH,DMF,80'C k 2 N -conc. HCI OH H OH To a cooled solution (0 OC) of compound 1 (0.190g, 0.72mmol) in anhydrous DMF (10mL) was added 0.028g, 0.72mmol) with stirring. The solution was allowed to warm to room temperature and stir for Compound 2 (0.275g, 0.73mmol) was added at room temperature and the mixture was heated at 60 °C for The solution was evaporated and the reside was partioned between ethyl acetate (150mL) and water WO 97/27180 PCT/US97/01610 179 The organic layer was washed with water (2 x brine (25mL), dried (MgSO 4 filtered, and evaporated to give a grey oil. The oil was purified by column chromatography: hexane/ethyl acetate (60/40) to give 0.23g of the acetonide protected product.
The acetonide (0.185g, 0.29mmol) was dissolved in isopropanol (10mL) and treated with conc. HC1 at room temperature. After 1.5h., the solution was adjusted to pH 11 with 3.0N NaOH and then concentrated.
The aqueous solution was extracted with ethyl acetate (3 x75mL). The ethyl acetate was dried (MgS04) and evaporatated to give a clear film. The crude product was purified by column chromatography: hexane/ethyl acetate (45/55) to give the product as a white solid (0.090g, Preparative HPLC on chiral phase (isopropanol-hexane gradient) yielded the desired diastereomer 3 (10mg) along with a 1:1 mixture of the desired diastereomer and an additional epimer MS: M+1= 603 H NMR (chloroform-d) 1.80(m, 6H); 2.50(m,1H); 2.60(m, 2H); 3.0(m,8H); 3.60(m,lH); 3.70(m,lH); 3.95(m,lH); 4.25(m,lH); 5.30(m,lH); 6.00(m,lH); 7.05(m,4H); 7.25(m,15H).
WO 97/27180 PCT/US97/01610 180 Synthesis of Compound 8 Ph 1 NaH, (s)-epichlorohydnn
DMF
Ph 0 2 Allyl lactam 1 (443 mg, 2.06 mmol) was dissolved in DMF (2 mL) and to this solution was added sodium hydride (2.2 mmol). The reaction mixture was stirred at room temperature for 1 hr after which (s)-epichlorohydrin (172 ul, 2.2 mmol) was added neat. The reaction was stirred at room temperature for 4 hr, diluted with water (20 mL) and extracted with ethyl acetate. The organic layer was then washed with water, brine and dried (MgSO 4 and filtered. Concentration in vacuo afforded crude epoxide product 2 which was used without further purification.
B.
Ph 0 1
HN
O NH 2 x Ph .I OH i-PrOH 3 Lactam epoxide 1 (180 mg, 0.66 mmol) and decahydroisoquinoline 2 (160 mg, 0.66 mmol) were heated WO 97/27180 PCT/US97/01610 181 to 80 °C in isopropanol. After three hours the reaction was cooled to 25 oC and stirred for 48 hours at room temperature. The reaction was then concentrated in vacuo. Purified by silica gel chromatography, eluting with 25 EtOAc Hexanes, providing 90 mg (90% pure by HPLC) of desired product 3.
Example 26 Synthesis of Compound 9
A.
Ph
N
Boc N O OH NBoc H O 2 \N HN i-PrOH, 75"C 0 t Bu 3 tu The Boc protected piperazine 1 (21.4 mg, 0.081 mmol), was dissloved in 1.5 mL of i-PrOH. This was followed by the addition of the lactam epoxide 2 (18.3 mg, 0.068 mmol). The reaction vessel was then fitted with a reflux condenser and heated to 75 °C for 16 hours. TIC indicated complete consumption of both starting materials and formation of a new material. The reaction was then cooled to 25 °C and concentrated in vacuo. The complete consumption of epoxide was confirmed by both tlc and H NMR. The crude addition product was then used without further purification.
WO 97/27180 PCT/US97/01610 182
B.
9 9 OH W 1) HCVdioxaneN 2) Picolyl Chloride-HC SNH TEA, DMF 24hr 0 NH 1 tBu 2 t Bu The Boc protected piperazine addition product 1 from the previous step was stirred for 2 hours in 1.0 mL of 4N HCl/dioxane. This was followed by concentration in vacuo. The crude solid was then dissolved in 10 mL of
CH
2 C1 2 and washed by 2 x 10 mL of each saturated aqueous sodium bicarbonate and saturated aqueous brine. The combined organic portions were then dried over MgSO 4 filtred and concentrated in vacuo to provide the freebase of the desired intermediate. The crude amine was then dissolved in 1.0 mL of DMF at 25 This was followed by the addition of the hydrochloride salt of 3-picolyl chloride (0.081 mmol). After stirring minutes triethylamine (300 uL, mmol) was added. The reaction was then stirred for 36 hours the reaction was quenched by the addition of 1.0 mL of saturated aqueous sodium bicarbonate. The reaction mixture was then diluted by the addition of 10 mL of diethyl ether and washed by 2 x 10 mL of each saturated aqueous sodium bicarbonate and saturated aqueous brine. The combined organic portions were then dried over MgSO 4 filtered and concentrated in vacuo to provide the crude product.
Purification of the crude solid was carried out by silica gel chromatography (1000 uM SiO 2 prep. plate) eluting with 20 MeOH/CH 2 C1 2 This provided 3.1 mg of the desired product 2, with 96 purity by HPLC. The WO 97/27180 PCT/US97/01610 183 overall yield for addition, deprotection of N-Boc and coupling with 3-picolyl chloride was 9 Example 27 Synthesis of Compound 3
A.
I) NaH,DMF Ph o2) 1 2 Allyl urea 1 (195.2 mg, 0.09 mmol) was dissolved in mL of DMF and cooled to 0 This was followed by the addition of NaH (54 mg, 1.0 mmol). The glycidyl tosylate (410 mg, mmol) was then added as a solid. The reaction was stirred for 4 hours at 25 °C and then quenched by the addition of 4 mL of saturated aqueous sodium bicarbonate. The reaction was then extracted by mL of Et 2 0. The organic layer was then washed by mL of saturated aqueous sodium bicarbonate and 2 x mL of saturated brine. The combined organic portions were then dried over MgSO4, filtered and concentrated in vacuo to provide the desired epoxide 2 (180 mg, 73 yield). The epoxide was then used without further purification.
WO 97/27180 PCT/US97/01610 184 Ph H 2 O i-PrOH, 75*C 18 hr O
NH
I
t Bu 1 Boc OH OH
Q/NH
3 t Bu Piperazine 1 (25.7 mg mmol) and epoxide 2 (22.6 mg, mmol) were heated to 75 °C in 1.5 mL of i-PrOH for 18 hours. After cooling to 25 °C the crude reaction mixture was concentrated in vacuo. Complete consumption of the epoxide was apparent by both tlc and 1H NMR.
C.
1 N-Boc OH fN' o 0 oNH 1 tBu 1)HCI/dioxane OH r N N 2) Picolyl Chloride-HCI 0 TEA, DMF 24 hr O NH t Bu 2 The Boc protected piperazine 1 from the previous step was stirred for 1.5 hours in 1.0 mL of 4 N HC1 in dioxane. This was followed by concentration in vacuo.
The crude hydrochloride salt was then dissolved in mL of CH 2 C1 2 and washed by 10 mL of both saturated sodium bicarbonate and saturated brine. The organic portion was then dried over MgSO 4 filtered and concentrated in vacuo. The free amine was then taken up in 1 mL of DMF. This was followed by the addition of 3-picolyl chloride HCI salt (50 mg, mmol) and WO 97/27180 PCT/US97/01610 185 triethyl amine (300 uL), respectively. The reaction was then stirred at 25 oC for 30 hours. The reaction was then quenched by the addition of 2 mL of saturated sodium bicarbonate and diluted by 10 mL of Et 2 0. The organic portion was then washed by 10 mL of saturated sodium bicarbonate and 2 X 10 mL of saturated brine.
The combined organic portions were then dried over MgSO 4 filtered and and concentrated in vacuo. The crude material was purified by silica gel chromatography (1000 uM prep. plate) eluting with 3:1,
CH
2 Cl 2 :MeOH to provide 8.8 mg of the desired product 2.
The overall yield for addition, deprotection of the N- Boc and reaction with 3-picolyl chloride was 19.3%.
Example 28 Synthesis of Compound 62 1) NaH, epichlorohydrin K, DMF
OH
H
H NH N N 2) Decahydroisoquinoline o isopropanol, heat 1 2 H The THF lactam 1 (0.4 mmol) was dissolved in dry DMF at 0 °C and to this solution was added sodium hydride (0.47 mmol). After 30 min of stirring, epichlorohydrin (0.47 mmol) was added and the reaction was allowed to warm to room temperature and stir overnight. The reaction was then diluted with water and extracted with ethyl acetate. The organic layer was washed sequentially with 0.5N HC1, saturated NaHCO 3 and brine, followed by drying (MgSO 4 filtration and WO 97/27180 PCT/US97/01610 186 concentration in vacuo to yield product (118 mg, crude) which was used as is. The lactam-epoxide (0.4 mmol, crude) was dissolved in isopropanol (2 mL), and to this solution was added decahydroisoquinoline t-butylamide (0.7 mmol). The mixture was then heated to 80 °C and stirred overnight. The reaction mixture was cooled and concentrated to dryness in vacuo, the residue of which was applied to a preperative TLC plate and eluted with 100% ethyl acetate to yield pure product (88 mg, 42%) as a mixture of diastereomers.
Example 29 Synthesis of Compound 92
A.
1. LDA/THF/-78iC BC 2. H
H
-78 0
C---RT
A stirred, cooled (-78 *C solution of 1.4 g mmol) of pyrrolidinone in 35 mL of anhydrous tetrahydrofuran was treated in a dropwise fashion with 3.6 mL (7.2 mmoL) of lithium diisopropylamide. The resultant solution was stirred for 70 min, and subsequently treated with 0.57 mL (6.0 mmoL) of 3pyridine carboxaldehyde. The homogenous solution was allowed to ambiently warm to room temperature and stirring was continued overnight. The reaction mixture was diluted with 400 mL of dichloromethane, washed once WO 97/27180 PCT/US97/01610 187 with 150 mL of water, dried (magnesium sulfate), filtered, concentrated, and purified on silica gel using 3:1 ethyl acetate/hexanes as the eluent, affording 0.6 g of the desired compound as a golden oil which solidified upon standing.
H NMR (d6-DMSO, 400MHz) 8.65 1H); 8.47 2H); 7.83 J 8.0 Hz, 1H); 7.41 1H); 7.23 7.03 J 2.7 Hz, 1H); 3.96 1H); 3.07 1H); 2.89 2.65 series of m, 3H M+H (265.2).
B.
H
2
N
H
CH
3 OH RT h H A vigorously stirred suspension of 330 mg 1.25 mmoL of eneamide and 80 mg of 10% palladium on carbon (Degussa) in 12mL of anhydrous methanol was hydrogenated (Hydrogen balloon) for 1 h. The mixture was diluted with 100 mL of methanol, carefully filtered, concentrated, and purified on silica gel using ethyl acetate as the eluent, affording 295 mg of an isomeric mixture of the desired compounds as a golden oil which solidified upon standing.
H NMR (d6-DMSO, 400MHz) 8.36 2H); 7.88 1H); 7.56 J 7.9 Hz, 1H); 7.27 7.12 7H); 3.66 1H); 2.96 2.37 (series of m, 7H). M+H (267.2); M+Na (289.2) WO 97/27180 WO 972718PCTIJS97/0 1610 -188-
C.
KI~1) NaH, DMF, 80*C cocHItasisomer1 NH cis-isomer 2 The lactam obtained above was coupled to the corresponding epoxide according to the protocol used for Example 24. The final purification was performed on silica gel 2M ammonia-methanol in dichioromethane) to give the cis- and the trans-lactam diastereomers each as white solids.
trans-isomer 1: Rf: 0.20 1HNMR (CDCl3,. 400MHz) l.62(2H,m), 1.86(4H,m), 2.19(1H,m), 2.63(2H-,m), 2.78- 3.10(8H,m), 3.65(lH,m), 3.75(lH,bt), 3.95(1H,t), 4.27(lH,t), 5.24(1H,m), 6.32(lH,d), 7 7 4 (14H-,m), 8.22(lH,s), 8.34(J.H,s). M+H (604) cis-isomer 2: Rf: 0.18. 1 H NMR (CDCl3,. 400MHz) 1.60(2H,m), 1.95(2H,m), 2.19(lH,dd), WO 97/27180 WO 9727180PCT[US97/01610 -189 2.48(lH,dd) 2.60(1H,m), 2.8-3.05(5H,m) 3.10(1I-,dd), 3.26(lH,dl), 3.60(1H,M), 3.78(lH,m), 3.99(l-,m), 4.15(lH,bs), 4.24(1H,t), 5.24(1H,m), 6.18(lI-,d), 7.02(2H,d), 7-7.3(10H,m), 7.41(1H,d), 8.25(1H,s), 8.40(lH,d). M+H (604) Example NH+ 0 NH CH 3 CN C N N N2 DIEA, 70 0
CH
12 3 The iodolactam 1 (0.43 mmol) was dissolved in dry acetonitrile in a high pressure tube and to this solution was added diisopropylethylamine (Pierce, 0.65 nunol) followed by aniline 2 (Aldrich, 0.47 mmol) The tube was sealed and the reaction heated to 70 0 C with stirring overnight. The reaction was cooled to ambient temperature, solvent removed in vacuo, and the residue taken up in ethyl acetate/water. The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was purified by flash silica gel chromatography eluting with 1:1 ethyl acetate/hexanes to give 61 mg of product 3; TLC Rf= 0.29 (1:1 ethyl acetate/hexanes); HPLC Rt 12.6 min MALDI-TOF MS m/z 267 WO 97/27180 PCT/US97/01610 190 Example 31
A.
Ph h Ph O 0CH13 LDA NC .Br NC 0 THF 0
O
1 2 PMB lactam 1 (1.5 g, 5.07 mmol) was dissolved in THF (12 mL), cooled to -78 and to this solution was added LDA (6.6 mmol 1.3 over 7 minutes to give a greenish-brown anion. The reaction mixture was stirred at -78 °C for 55 minutes after which a solution of-bromoacetonitrile (400 ul, 0.75 mmol, 1.1 eq.) was added over 2 minutes while keeping the internal reaction temperature at <-65 oC. The reaction was stirred at -78 °C for 2 hours, then warmed to room temperature and stirred for an additional 16 hours.
The reaction was cooled to -50 °C and quenched with saturated ammonium chloride solution. The reaction was partitioned between ethyl acetate and a saturated bicarbonate solution. The aqueous layer was extracted with ethyl acetate. The combined organic layers were then washed with water, brine and dried (MgSO4) and filtered. Concentration in vacuo afforded 1.6g of crude material, which was purified by silica gel chromatography to give 640 mg of the desired material 2.
H NMR (CDCl 3 d 7.31 3H), 7.18 2 7.09 (d, 2H), 6.90 2H), 5.08 1H), 3.92 1H), 3.81 (s, WO 97/27180 PCT/US97/01610 191 3H), 3.70 1H), 2.92 (dd, 1H), 2.72 2.55 (dd,1H), 2.42 2.19 1.81 1H).
B.
Ph Ph O C xi (NH 4 2 Ce(NO )6 NC NC NH 1 CH 3 CN:water 2 0 9:1 0 1 2 PMB lactam 1 (640mg, 1.9 mmol) was dissolved in CH 3
CN
(9 mL). 1 mL of water was added followed by 3.1 g of cerium ammonium nitrate. The reaction went from dark amber to light orange within 5 minutes and was stirred at room temperature for 18 hours. The reaction was concentrated in vacuo and the residue was partitioned between ethyl acetate and a saturated bicarbonate solution. The.aqueous layer was extracted with ethyl acetate. The combined organic layers were then washed with saturated bicarbonate solution, water, brine, dried (MgSO4) and filtered. Concentration in vacuo afforded 590 mg of crude material, which was purified by silica gel chromatography (9:1 CH 2 Cl 2 EtOAc) to give 285 mg of the desired material 2. HPLC suggests 2 diastereomers, retention time 9.95 min.(major) and 10.17 min. (minor).
H NMR (CDC1 3 d 7.37 2H), 7.28 1 7.20 (m, 2H), 5.74 (br s, 1H), 3.95 1H), 2.85 (dd, 1H), 2.79-2.65 3H), 2.55 (dd, 1H), 2.27 (m,2H).
WO 97/27180 PCT/US97/01610 192 Example 32
A.
SMe 1)LDA,THF OMe N 2 1 2 3 The PMB lactam 1 (0.46 mmol) was dissolved in dry THF at -78 OC and to this solution was added lithium diisopropylamide (Aldrich, 1.5 M in cyclohexane, 0.65 mmol). The solution was stirred for 15 minutes at -78 oC and 4-(Chloromethyl)-3,5-dimethylisoxazole 2 (Acros Organics, 0.56 mmol) was added. The cooling bath was removed and the solution warmed to room temperature and stirred overnight. The reaction was diluted with water and extracted with ethyl acetate.
The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was purified by flash silica gel chromatography eluting with 10% diethyl ether/dichloromethane to give 53 mg of product 3 as a mixture of diastereomers.
WO 97/27180 PCT/US97/01610 193
B.
OMe SN 7:3 CH 3
CN/H
2 0 O
O
1 2 Lactam 1 (0.13 mmol) was dissolved in 7:3 acetonitrile/water. Ceric ammonium nitrate (Aldrich, 0.26 mmol) was added and the mixture was stirred at ambient temperature until the starting material was no longer evident by TLC. Acetonitrile was removed in vacuo, and the residue taken up in ethyl acetate/water.
The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was purified by flash silica gel chromatography eluting with 8% MeOH in dichloromethane to give 21 mg of product 2; TLC Rf 0.47 (8% MeOH/CH 2 Cl 2 WO 97/27180 PCT/US97/01610 194 Example 33
A.
N PMB 1)LDA
N
PMB 2) p-formaldehyde
PMB
O 3) NBS, PPh3 0 1 2 Lactam 1 (1.43 mg, 4.86 mmol) was dissolved in anhydrous THF (25 mL) and cooled to -78 oC. This was followed by the addition of 3.9 mL of LDA (5.83 mmol, 1.2 The anion solution was stirred at -78 °C for minutes and then cannulated into a -78 °C solution of p-formaldehyde (437 mg) in 25 mL of THF, washing with 1 mL of THF. The reaction was warmed to room temperature over 4 hr and stirred overnight. The reaction was quenched by the addition of 10 mL of a saturated sodium bicarbonate, and concentrated in vacuo to remove the THF. The crude reaction mixture was partitioned between ethyl acetate and saturated sodium bicarbonate. The aqueous layer was extracted with ethyl acetate. The combined organic layers was then washed with water, brine and purified by silica gel chromatography (gradient of 50 to 75 ethyl acetate: hexanes), to provide 584 mg of the desired alcohol, as well as 265 mg of recovered starting material.
WO 97/27180 PCT/US97/01610 195 The alcohol (316mg, 0.979 mmol) was then dissolved in 3 mis of CH 2 C12 and added to a 0 OC solution of triphenyl phosphine (734 mg, 2.8 EQ.) and NBS (534 mg, 3 EQ.) in 3 mls of CH 2 Cl 2 After 1 hour the reaction was quenched by the addition of 10 mL of Et 2 0. The organic layer was then filtered and the filtrate washed with saturated sodium bicarbonate, brine, dried (MgSO 4 and filtered. Concentration in vacuo afforded the crude product which was purified by silica gel chromatography
(CH
2 C1 2 to provide 151 mg (40% of the bromide.
The bromide (87.2 mg, 0.28 mmol) was dissolved in 2 mL of benzene and treated with imidazole (46mg, 3 EQ.).
After heating to 125 °C for 20 hours the reaction was cooled to 25 °C and concentrated in vacuo. The crude product which was purified by silica gel chromatography MeOH/CH 2 C12), to provide the addition product and the elimination product in a 50 yield.
B.
1)CAN N PMB 2) imidazole NN _N NH 0 0 1 2 The lactam 1 (621 mg, 2.02 mmol) was dissolved in 7 mL acetonitrile, followed by the addition of H 2 0 (3 mL).
This was followed by the addition of CAN, 3.32 g (6.06 mmol, 3 The reaction was stirred at 25 oC for 1 WO 97/27180 PCTIUS97/01610 196 hour. After concentrating the reaction in vacuo, the crude material was resuspended in ethyl acetate and washed with saturated sodium bicarbonate, brine, dried (MgSO 4 and filtered. Concentration in vacuo afforded the crude product which was purified by silica gel chromatography methanol:CH 2 Cl 2 to procide the desired unprotected lactam (122 mg, 32 The cxa,-unsaturated lactam (55 mg, 0.29 mmol) was then heated to 130 °C in 2 mL of benzene containing imidazole (30 mg, 0.44 mmol) for 24 hours. After cooling to 25 the reaction mixture was concentrate in vacuo. The crude material was purified by silica gel chromatography, eluting with 5% methanol:CH 2 Cl 2 to provide 46.7 mg of the desired addition product (63 as well as 15.7 mg of recovered starting olefin (29 Example 34 I NH CH 3
CN
N H N NH O DIEA, 700C 1 2 3 The iodolactam 1 (0.45 mmol) was dissolved in dry acetonitrile in a high pressure tube and to this solution was added diisopropylethylamine (Pierce, 1.35 mmol) followed by indoline 2 (Aldrich, 0.54 mmol). The WO 97/27180 PCT/US97/01610 197 tube was sealed and the reaction heated to 70 oC with stirring overnight. The reaction was cooled to ambient temperature, solvent removed in vacuo, and the residue taken up in ethyl acetate/water. The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was purified by flash silica gel chromatography eluting with ethyl acetate to give 113 mg of product 3; TLC Rf 0.39 (ethyl acetate); HPLC Rt 13.1 min MALDI-TOF MS m/z 293 Example
A.
Ph OMe Ph EtNO 2 Oe PhNCO 0 Et 3
N
1 2 In an oven-dried 100 mL round-bottomed flask, the vinyl sulfone PMB lactam 1 (1.2126 g, 2.55 mmol) was dissolved in 50 mL of C 6 Hg. Phenyl isocyanate (2.0 mL, 18.4 mmol) was added via syringe followed by the dropwise addition of nitroethane (0.4 mL, 5.56 mmol).
Triethylamine (2.0 mL, 14.3 mmol) was added dropwise.
The solution was refluxed for 15 minutes and cooled. A white solid precipitated during the heating period.
The mixture was cooled, poured into water and extracted WO 97/27180 PCTIUS97/01610 198 with CH 2 C12. The organic extract was dried (MgSO 4 and evaporated in vacuo to afford a brown oil that was chromatographed to afford the isoxazole PMB lactam 2 (901 mg, 90%) as a light yellow oil.
B.
PhOMe Ph -o CAN CtH3CN-H20
H
0 1 2 In a 25 mL round-bottomed flask, isoxazole PMB lactam 1 (900 mg, 2.30 mmol) was dissolved in 14 mL of
CH
3
CN-H
2 0. Ceric ammonium nitrate (3.607 g, 6.58 mmol) was added forming a dark orange solution. The mixture was stirred until the starting material was no longer evident by TLC.(10% EtOAc/CH 2 Cl 2 The light yellow solution was diluted with CH 2 C12 and washed with water.
The organic layer was separated, dried (MgS04), and evaporated in vacuo to afford a brownish-red oil that was chromatographed (10% EtOAc/CH 2 Cl 2 to produce the lactam 2 (300.3 mg, 48%) as a colorless oil.
WO 97/27180 PCT/US97/01610 199 Example 36 NH a NH DCH 3 CN NH Y Nr DIEA, 700C o I 0 1 2 3 The iodolactam 1 (0.78 mmol) was dissolved in dry acetonitrile in a high pressure tube and to this solution was added diisopropylethylamine (Pierce, 2.35 mmol) followed by N-methylaniline 2 (Aldrich, 0.94 mmol). The tube was sealed and the reaction heated to oC with stirring overnight. The reaction was cooled to ambient temperature, solvent removed in vacuo, and the residue taken up in ethyl acetate/water. The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was purified by flash silica gel chromatography eluting with 2:1 ethyl acetate/hexanes to give 134 mg of product 3; TLC Rf 0.24 (2:1 ethyl acetate/hexanes); HPLC Rt 12.7 min MALDI- TOF MS m/z 282 WO 97/27180 ]PCTIUS97/01610 200 Examile 37
A.
H
I" 0 0 dNaH, Cul dioxane 100 0
C
1 2 NaH (0.96 g, 40 mmol) was suspended into 20 mL of dioxane. This was followed by the addition of diethyl malonate (4.6 mL, 40 mmol), then phenyl iodide (2.2 mL, mmol) and finally copper iodide (7.6g, 40 mmol).
The reaction was then heated to 100 oC for 14 hours.
The reaction was then quenched with water and diluted with ethyl acetate, the organic layer was washed with water and saturated NaC1, dried (MgSO 4 and concentrated in vacuo. The crude product was further purified by MPLC (SiO 2 eluting with 4:1, toluene: ethyl acetate to provide 1.21 g of product (29 isolated yield).
B.
CsC03 MeCN WO 97/27180 PCT/US97/01610 201 The alkylated malonic ester 227 mg, 1.09 mmol) was stirred for 14 hours in acetonitrile (2.5 mL) containing; cesium carbonate (710 mg, 2.18 mmol) and the bromide (516 mg, 1.31 mmol). The reaction was then concentrated to dryness in vacuo. After re-suspension of the reaction mixture in ethyl acetate, the reaction mixture was washed with water, saturated NaHCO 3 and saturated NaCl, dried (MgSO 4 and concentrated in vacuo. The crude product was further purified by MPLC (SiO 2 to provide 200 mg of the desired product 35.2 yield).
C.
EtOOC
H
2 (approx. 1 ATM) EtO? N '-NH Pd/C, HCI, EtOH Et S 1 2 To the malonate 200 mg) in ethanol (3 mL) was added concentrated HC1 (100uL) and an excess of 5% Pd C (approx. 50 mg). The reaction was then fitted with a balloon of H 2 and hydrogenated for 14 hours. After purging the reaction mixture of H2, triethylamine (1 mL, 7 mmol, excess) and an excess of solid NaHCO3 was added. After stirring for 30 minutes the reaction was filtered and concentrated in vacuo. The yellow oil was WO 97/27180 PCT/US97/01610 202 then re-dissolved in ethyl acetate and the reaction mixture was washed with water, saturated NaHCO 3 and saturated NaCl, dried (MgS04) and concentrated in vacuo to provide the desired product. The H NMR was consistent with the desired material.
Example 38
A.
Ph, Ph OMe OMe 1)LDA,THF 2) propargyl bromide 0 0 1 2 In an oven-dried 25 mL round-bottomed flask, the PMBlactam 1 (563.7 mg, 2.75 mmol) was dissolved in 10 mL of THF. The solution was cooled to -78 °C and 1.5M LDA mL, 3.00 mmol) was added dropwise via syringe producing the yellow color of the enolate. The solution was stirred for 15 minutes at -78 oC and propargyl bromide (310 uL, 3.48 mmol) was added dissipating the yellow color. The cooling bath was removed and the solution was warmed to room temperature and stirred overnight. The solution was poured into IN HC1 and extracted with CH 2 C1 2 The organic extracts were combined and washed with saturated aqueous NaHCO 3 The organic layer was separated, dried (MgSO 4 and evaporated in vacuo to afford a brown oil that was chromatographed (90% CH 2 Cl 2 /hexane) to produce the propargyl lactam 2 (577 mg, 86%) as a colorless oil.
WO 97/27180 PCT/US97/01610 203
CAN
CH3CN20 Ph
NH
In a 25 mL round-bottomed flask, propargyl PMB lactam 1 (358.2 mg, 1.08 mmol) was dissolved in 6 mL of
CH
3
CN-H
2 0. Ceric ammonium nitrate (1.321 g, 2.41 mmol) was added forming a dark orange solution. The mixture was stirred until the starting material was no longer evident by TLC (10% EtOAc/CH 2 Cl 2 The light yellow solution.was diluted with EtOAc and washed with water.
The organic layer was separated, dried (MgSO 4 and evaporated in vacuo to afford a yellow oil that was chromatographed (10% EtOAc/CH 2 Cl 2 to produce the propargyl lactam 2 (145 mg, 63%) as a colorless oil.
Example 39.
NH 0 1
CH
3
CN
DIEA, 70 0
C
The iodolactam 1 (1.38 mmol) was dissolved in dry acetonitrile in a high pressure tube and to this WO 97/27180 PCT/US97/01610 204 solution was added diisopropylethylamine (Pierce, 4.15 mmol) followed by tetrahydroquinoline 2 (Aldrich, 1.66 mmol). The tube was sealed and the reaction heated to OC with stirring overnight. The reaction was cooled to ambient temperature, solvent removed in vacuo, and the residue taken up in ethyl acetate/water. The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was purified by flash silica gel chromatography eluting with 1:1 ethyl a-cetate/hexanes to give 233 mg of product 3; TLC Rf 0.21 (1:1 ethyl acetate/hexanes); HPLC Rt 14.0 min MALDI- TOF MS m/z 307 Examle
A.
Ph OMe Ph OMe 1)PhSCI -N 2) MCPBA PhO2S
N
S3)DBU
O
1 2 In an oven-dried 250 mL round-bottomed flask, Nchlorosuccinimide (2.5177 g, 18.9 mmol) was dissolved in 75 mL of CH 2 Cl 2 The solution was cooled to 0 °C and thiophenol (1.90 mL, 18.5 mmol) was added dropwise via syringe causing an immediate formation of a yellow color and an exotherm. The orange solution of PhSC1 was stirred for 30 minutes at room temperature and a WO 97/27180 PCT/US97/01610 205 solution of the allyl lactam 1 (6.156 g, 18.4 mmol) was added dropwise dissipating the orange color. The light yellow solution was stirred for two hours and the solvent was removed in vacuo. CC14 was added to the yellow oil that remained and the undissolved succinimide was removed by filtration. The filtrate was evaporated in vacuo to afford the diastereomeric chlorosulfides as a yellow oil that was chromatographed
(CH
2 C12) rapidly to remove low Rf impurities. The two highest Rf spots were the chlorosulfide diasteromers.
The purified mixture of chlorosulfides was dissolved in
CH
2 C12 and m-chloroperbenzoic acid (2.0 g, 11.6 mmol) was added with cooling from an ice-bath. The mixture was stirred for 10 minutes and filtered. The filtrate was evaporated in vacuo to afford a yellow oil (8.125 g, 86%) that produced two low Rf spots (CH 2 C1 2 using thin-layer chromatography for the two chlorosulfone diastereomers. The oil was redissolved in CH2C12 and DBU (2.7 mL, 18.1 mmol) was added dropwise at room temperature. The solution was heated for 15 minutes causing the solution to turn dark yellow. The solution was cooled and the solvent was evaporated in vacuo.
The residue was chromatographed (CH 2 C1 2 to afford the pure vinyl sulfone 2 (4.805 g, 55%) as a colorless oil.
B.
Ph OMe Ph OMe TMSCHN S PhO2S BuLi
THF
WO 97/27180 PCT/US97/01610 206 In an oven-dried 25 mL round-bottomed flask, trimethylsilyl diazomethane (140 uL, 0.280 mmol) was dissolved in 5 mL of THF. The bright yellow solution was cooled to -78 °C and n-BuLi (320 uL, 480 mmol) was added. In a separate oven-dried 25 mL round-bottomed flask, the vinyl sulfone PMB lactam 1 (108 mg, 0.227 mmol) was dissolved in 5 mL of THF and added dropwise via syringe at -78 OC to the lithiate solution. The resulting solution was stirred for 1 hour at -78 °C and then two hours at 0 The mixture was acidified with IN HC1 and extracted with CH 2 Cl 2 The organic extract was dried (MgSO 4 and evaporated in vacuo to afford a cloudy, colorless oil that was chromatographed EtOAc/CH 2 Cl 2 to produce the TMS pyrazole PMB lactam 2 (88.4 mg, 87%) as a clear, colorless oil.
C.
MS P OMe TBAF POMe CH3CN
H
O
1 2 In an oven-dried 25 mL round-bottomed flask, the TMS pyrazole PMB lactam 1 (1.1345 g, 2.53 mmol) was dissolved in 110 mL of 91% CH 3
CN/H
2 0.
Tetrabutylammonium fluoride (2.7 mL of a 1.OM solution in THF, 2.70 mmol) was added dropwise via syringe. The reaction was refluxed for 48 hours and cooled. The solvent was evaporated in vacuo and the residue was WO 97/27180 PCT/US97/01610 207 dissolved in CH 2 C1 2 The organic solution was washed with 1N HC1 solution, dried (MgSO 4 and evaporated in vacuo to afford a yellow oil that was chromatographed EtOAc/CH 2 C12) to afford the pyrazole (688 mg, 72%) as a light yellow oil.
D.
S/ 1) NaH OPh\ 2) Me02CCI Me-O 0 0 1 2 In an oven-dried 100 mL round-bottomed flask, the pyrazole PMB lactam 1 (588 mg, 1.57 mmol) was dissolved in 25 mL of THF. NaH (50 mg of a 60% dispersion in mineral oil, 2.08 mmol) was added. Gas evolution was observed. Methyl chloroformate (140 uL, 1.81 mmol) was added and the reaction was stirred at room temperature overnight. The mixture was acidified with IN HC1 and extracted with CH2C1 2 The organic extract was dried (MgSO 4 and evaporated in vacuo to afford the pyrazole carbamate PMB lactam 2 (588 mg, 87%) as a light yellow oil.
E.
Ph\ OMe Ph O CAN O Me- 70% acetonitrile/water Me-O 'H WO 97/27180 PCT/US97/01610 208 In an oven-dried 100 mL round-bottomed flask, the pyrazole carbamate PMB lactam 1 (577 mg, 1.33 mmol) was dissolved in 30 mL of 70% CH 3
CN-H
2 0. Ceric ammonium nitrate (2.5123 g, 4.58 mmol) was added. The orange solution was stirred at room temperature until the starting material was no longer evident by TLC (1 hr).
The light yellow solution was poured into water and extracted with EtOAc. The organic extract was dried (MgSO 4 and evaporated in vacuo to afford the pyrazole carbamate lactam 2 (228 mg, 55%) as a clear, colorless oil.
Example 41
A.
Ph OMe H Ph OMe Bu 3 SnN 3 N xylene
N
0 205°C 1 2 In a heavy-walled screw-top test tube, the propargyl lactam 1 (1.111 g, 3.33 mmol) was dissolved in 7 mL of xylene. Tributyltin azide (1.965 g, 5.92 mmol) was added, the tube was sealed and heated to 205 °C overnight. The dark brown solution was cooled and directly chromatographed using a gradient from CH 2 C12 to 50% EtOAc/CH 2
CI
2 to afford the triazole PMB lactam 2 (827 mg, 66%) as a light yellow oil.
WO 97/27180 PCT/US97/01610 209
B.
H Ph OMe Ph OMe M L. OMe N N 1) Na H ^N 2) PhCH 2 Br 0 1 2 In an oven-dried 100 mL round-bottomed flask, the triazole PMB lactam 1 (827 mg, 2.20-mmol) was dissolved in 40 mL of THF. NaH (124 mg of a 60% dispersion in mineral oil, 5.17 mmol) was added. Gas evolution was observed. Benzyl bromide (400 uL, 3.36 mmol) was added. The reaction was stirred at reflux until the starting material was not longer evident by thin-layer chromatography (50% EtOAc/CH 2 Cl 2 The mixture was acidified with 1N HC1 and extracted with CH 2 C1 2 The organic extract was dried (MgSO 4 evaporated in vacuo to afford a dark yellow residue that was chromatographed (20% EtOAc/CH 2 Cl 2 to produce the benzyl triazole PMB lactam 2 (740 mg, 72%) as a light yellow oil.
C.
Ph Ph Ph OMe Ph i <I70% CH 3
CN-H
2 0O NH 0 0 1 2 In an oven-dried 50 mL round-bottomed flask, the benzyl triazole PMB lactam 1 (740 mg, 1.59 mmol) was dissolved WO 97/27180 PCT/US97/01610 210 in 22 mL of 70% CH 3
CN-H
2 0. Ceric ammonium nitrate (2.1 g, 3.83 mmol) was added. The orange solution was stirred at room temperature until the starting material was no longer evident by TLC (1 hr). The mixture was poured into water and extracted with EtOAc. The organic extract was .dried (MgSO 4 and evaporated in vacuo to afford the benzyl triazole lactam 2 (336 mg, 61%) as a clear, colorless oil.
Example 42 1) LDA/THF 2) acetone N-BOC 3) Martin's rgt
H
0 CH2CI2 0 4) H 2 /10% Pd-C 1 MeOH 2 TFAICH2CI 2 BOC-lactam 1 (1.8 g, 6.6 mmol) was dissolved in THF mL) and cooled to -78 To this solution was added LDA (Aldrich, 1.5 M in cyclohexane, 5.3 mL, 7.9 mmol) via syringe over 10 minutes. After stirring for 60 min at -78 acetone (4.9 mL, 66 mmol) was added via syringe over 1 minute. The reaction was stirred for an additional 40 minutes before being quenched with 1N HC1 mL). Ethyl acetate (100 mL) was added and the layers were partitioned. The organic layer was washed with brine, dried over magnesium sulfate, filtered, and concentrated in vacuo to a yellow oil that slowly WO 97/27180 PCT/US97/01610 211 crystallized. The crude alcohol was dissolved in dichloromethane (50 mL) and Martin's sulfurane (Aldrich, 7.5 g, 11 mmol) was added in one portion.
The reaction was stirred for 36 h at room temperature before being concentrated in vacuo. Flash chromatography over silica gel (3:1 hexane:ethyl acetate) provided the alkene as a mixture of isomers.
The alkene, 10% Pd-C (1.0 and methanol (40 mL) were combined in a Parr bottle and pressurized to 50 psi of hydrogen gas. After 4 h of agitation, the reaction vessel was evacuated and filtered through a plug of Celite. The cake was washed with ethyl acetate (20 mL) and the combined filtrate was concentrated in vacuo to give the isopropyl BOC-lactam as a pale yellow oil.
The lactam was dissolved in dichloromethane (20 mL) and trifluoroacetic acid (10 mL) was added slowly. The reaction was stirred at room temperature for 24 h before being diluted with ethyl acetate (100 mL) and carefully neutralized with 10% sodium carbonate to pH 7. The layers were partitioned and the organic layer was dried over magnesium sulfate, filtered, and concentrated in vacuo. Flash chromatography over silica gel (3:1 ethyl acetate:hexane) gave the isopropyl lactam as a white powder. MS 240 (M+Na) WO 97/27180 PCT/FJS97/01610 212 Example 43 1. LDA/ THF/-78 0
C
_Boc 2 N O o 0 o
H
-78 0
RT
A stirred, cooled (-78 solution of 1.4 g (5.0 mmol) of pyrrolidinone in 35 mL of anhydrous tetrahydrofuran was treated in a dropwise fashion with 3.6 mL (7.2 mmoL) of lithium diisopropylamide. The resultant solution was stirred for 70 min, and subsequently treated with 0.57 mL (6.0 mmoL) of 3-pyridine carboxaldehyde. The homogenous solution was allowed to ambiently warm to RT, and stirring was continued overnight. The reaction mixture was diluted with 400 mL of dichloromethane, washed IX with 150 mL of water, dried (magnesium sulfate), filtered, concentrated, and purified on silica gel using 3:1 ethyl acetate/hexanes as the eluent affording 0.6 g of the desired compound as a golden oil which solidified upon standing. H NMR (d6-DMSO, 400MHz) 8.65 1H); 8.47 2H); 7.83 J 8.0 Hz, 1H); 7.41 1H); 7.23 5H); 7.03 J 2.7 Hz, 1H); 3.96 1H); 3.07 1H); 2.89 2.65 (series of m, 3H). M+H (265.2).
WO 97/27180 PCTIUS97/01610 213 Example 44 1) LDA pyridine-3-carboxaldehyde N Boc -780 C S2) H1/Pd-C NH
O
O
1 2 The first step of the sequence was performed as for Example 43. The olefin was carried forward as follows: step 2 A vigorously stirred suspension of 330 mg (1.25 mmoL) of eneamide and 80 mg of 10% palladium on carbon (Degussa) in 12mL of anhydrous methanol was hydrogenated (Hydrogen balloon) for 1 h. The mixture was diluted with 100 mL of methanol, carefully filtered, concentrated, and purified on silica gel using ethyl acetate as the eluent affording 295 mg of an isomeric mixture of the desired compounds as a golden oil which solidified upon standing. 1H NMR (d6-DMSO, 400MHz) d 8.36 2H); 7.88 1H); 7.56 J 7.9 Hz, 1H); 7.27 7.12 7H); 3.66 (m, 1H); 2.96 2.37 (series of m, 7H). M+H (267.2); M+Na (289.2) WO 97/27180 WO 9727180PCT/US97/01610 214 ExaMple 1) LDA pyridine-2-carboxaldchyde
N
-78 0
CN
1?00C 2) H 2 Pd-CH 00 1 2 The synthesis of the 2-pyridyl methylpyrrolidone was carried out as shown in-Example 44.
2NBOC 0 1) LDA pyndine-4-carboxaldehyde -7 8 0 C 2) H 2 /Pd-C
NH
0 The 4-pyridylmethylpyrrolidone was prepared following procedures outline for Example 44.
WO 97/27180 PCT/US97/01610 215 Example 47 1. (MeO) 2
POCH
2 COOtbu/ NaH 2. H2 Pd-C 3. LHS Boc-Phenylalaninal 0 X 4. TFA, neutralize X=O, N-Bn X N-Bn
A.
A solution of 5.06g (20 mmol, 1 equiv) of tert-Butyl- P,P-dimethylphosphonoacetate in 15 mL THF cooled to 0 oC was treated with 0.528g of NaH at 0 OC and then warmed up to room temperature for 30 min. Next, solution of 5.0g (20 mM, 1 equiv) of Boc-Phenylalaninal in 5 mL THF was added dropwise at 0°C and the reaction continued for 2 h. The crude product was diluted with ethyl acetate and partitioned with aqueous citric acid sodium bicarbonate organics collected and dried over magnesium sulfate. The product was then dissolved in 100 mL methanol, added 0.6g 10% Pd/C and hydrogenated at 25 psi overnight, and the desired compound purified on a silica column using 1/4 ethyl acetate/hexane. Yield 3.8g H NMR (CDCL 3 300 MHz) 5 (broad signals and conformational averaging) 7.20 5H), 4.46 3.79 0.5H), 3.72 0.5H), 2.80 0.5H), 2.46 0.5H), 2.27 1H), 1.78 1H), 1.50 1H), {1.44, 1.42, 1.41, 1.38} (all s, total 18H). Low resolution MS m/e 372.2 WO 97/27180 PCT/US97/01610 216
B.
A solution of 4.63 g (13.25 mmol, 1 equiv) of the above ester in 200 mL of THF was treated with 40 mL (39.75 mmol, 3 equiv) of IM lithium bis(trimethylsilyl) amide in THF at -78 oC. After 90 min at -78 OC, the solution was added 5.5g (13.25.mmol, 1 equiv) of N-benzyl-Nbis(iodoethane) in 10 mL of THF and the reaction continued for 6 hours during which it reached the room temperature. The reaction was quenched with 10% aqueous solution of citric acid and extracted to ethyl acetate, and the product treated with 1:1 DCM/TFA (40 mL) for 40 min, after which solvents were removed and the crude purified to homogeneity by RP HPLC with total yield of 14.2%. The resulting TFA salt was then neutralized with triethylamine, extracted between ethyl acetate/water, organics collected and dried, thus yielding a free base form of the spiropyrrolidone product which is used in subsequent coupling to the epoxide. 1H NMR (TFA salt, CDCL 3 300 MHz) 5 7.30 (m, 10H), 5.85 1H), 4.16 2H), 3.86 1H), 3.68 1H), 3.36 3H), 2.88 (dd, 1H), 2.62 (dd, 1H), 1.7-2.2 6H). Low resolution MS m/e 335.2 (M+H Example 48 Spirocycle X=O was synthesized according to bisalkylation protocol of Example 47 above except that bis-O-(iodoethyl) ether was used in reaction step B (1.26g, 3,87 mmol, 1 equiv). 1H NMR (d 6 -DMSO, 300 MHz) 7.79 1H), 7.22 5H), 3.73 3H), 3.24 (m, 2H), 2.88 (dd, 1H, J=4.8, 13.4), 2.57 (dd, 1H, J=8.4, 13.4), 2.03 1H), 1.76 1H), 1.55 2H), 1.22 WO 97/27180 PCT/US97/01610 217 1H), 1.01 1H). Low resolution MS m/e 246.2
(M+H
Example 49 1. (MeO) 2
POCH
2 COOtbu NaH 2. H2 Pd-C 3. LHS Boc-Phenylalaninal x=cii X=Cft Spirocycle X=CH 2
A.
A solution of 1.36g (3.88 mmol, 1 equiv) of the ester from Example 47 step A in 5 mL of THF was cooled to 78 °C and treated with 9.32 mL (9.32 mmol, 2.4 equiv) of 1M lithium bis(trimethylsilyl) amide in THF. After 1 h at -78 0.992g (4.27 mmol, 1.1 equiv) of iodochloropentane was added and the reaction allowed to progress at -15 °C for 1 h, quenched with 10% aqueous citric acid, and extracted to ethyl acetate, resulting in 1.60g of product. Low resolution MS m/e 476.2 (M+Na
B.
A solution of 1.6g (3.53 mmol, 1 equiv) of the above chloride in 30 mL acetone was treated with 5.29 g (35.3 mmol, 10 equiv) of NaI and refluxed overnight. Solvents were then removed and the residue partitioned between ethyl acetate/water. Organics were dried with magnesium WO 97/27180 PCT/US97/01610 218 sulfate and purified on silica gel using 1/3 ethyl acetate/hexane resulting in 1.2 g of the desired iodide (62.4% yield after chromatography).
1H NMR (CDCL 3 300 MHz) 5 7.20 5H), 4.38 1H), 3.79 1H), 3.13 2H, 2.73 2H), 2.25 1H), 1.76 2H), 1.43 9H), 1.38 9H), 1.2- 1.7 7H). Low resolution MS m/e 568 (M+Na m/e 362.2 (M+H
C.
A solution 1.15g (2.1 mM, 1 equiv) of the above product in 20 mL of anhydrous THF was cooled to -78 OC and treated with 3.2 mL (3 mmol, 1.5 equiv) of 1M lithium bis(trimethylsilyl) amide in THF. The reaction was then allowed to warm up to room temperature, solvents removed and the crude product purified on preparative HPLC. H NMR (CDCL 3 300 MHz) 5 7.32 4H), 7.19 (d, 12H), 3.88 1H), 2.82 2H), 2.24 (dd, 1H), 1.2- 1.8 11H). Low resolution MS m/e 384.2 (M+Na m/e 362.2 (M+H Example Ni-B( 1) LDA/THF
N-
2) 2) .D CHO i N 3. H 2 Pd-C WO 97/27180 PCT/US97/01610 219
A.
The Boc-pyrrolidone (4.4 g, 16 mmol) was dissolved in THF (40 mL) and cooled to -78 To this solution was added LDA (Aldrich, 1.5 M in cyclohexane, 12.8 mL, 19 mmol) via syringe over 10 minutes. After stirring for min at -78 OC, 3-formyl-5,6-dihydro-2H-pyran
(US
Patent 4,532,337) (1.8 g, 16 mmol) in THF (5 mL) was added via syringe over 1 minute. The reaction was then allowed to reach room temperature and stir for 20 h before being quenched with saturated ammonium chloride mL). Ethyl acetate (50 mL) was added and the layers were partitioned. The organic layer was washed with brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography over silica gel (95:5 chloroform:methanol) to give dihydropyran lactam as a beige powder. MS 270 292 (M+Na)
B.
The dihydropyran obtained above (1.2 g, 4.4 mmol), Pd-C (0.2 and methanol (35 mL) were combined in a Parr bottle and pressurized to 50 psi of hydrogen gas.
After 3 h of agitation, the reaction vessel was evacuated and filtered through a plug of Celite. The cake was washed with methanol (20 mL) and the combined filtrate was concentrated in vacuo. Flash chromatography over silica gel (95:5 chloroform:methanol) gave the tetrahydropyran lactam 2 as a white powder. MS 274 296 (M+Na) WO 97/27180 PCT/US97/01610 220 Example 51 NBo
NH
A.
A solution of 2.6g (8.24 mmol, 1 equiv) of the allylpyrrolidone in 80 mL tetrahydrofuran and 25 mL water was cooled to 0 °C and treated with 5.29 g (24.7 mmol, 3 equiv) NaIO 4 followed by the addition of 838 mg of solution of osmium tetroxide in 2-methyl-2propanol. The reaction was continued for 2 h at room temperature, solvents removed and the residue partitioned between ethyl acetate and water. Ethyl acetate was then dried over MgSO 4 resulting in 3.0 g of the crude aldehyde.
H NMR (CDCL 3 300 MHz) 5 9.75 1H), 7.22 4.32 1H), 3.05 2H), 2.82 3H), 2.53 1H), 2.22 1H), 1.58 9H). Low resolution MS m/e 356.1 m/e 689.3 (2M+Na+).
B.
A solution of 2.88g of the above aldehyde in 10 mL methanol was cooled to 0 °C and sodium borohydride was added over 2 h, until all the starting material (Rf=0.55, Merck Kiselgel 60, 0.25 mm, 1:1 ethyl acetate/hexane) was consumed. The title compound had Rf=0.30 (same conditions). Solvents were then removed, and the residue was extracted between ethyl acetate and WO 97/27180 PCT/US97/01610 221 aqueous citric acid. Organic fractions were washed with water and dried over magnesium sulfate.
Purification on a silica column (1:1 ethyl acetate/hexane) afforded 1.5 g (57% yield) of the alcohol. Low resolution MS m/e 342.2 m/e 661.4 (2M+Na+)
C.
A solution of 0.46g (1.44 mmol, 1 equiv) of the above alcohol in 4 mL tetrahydrofuran was treated with 0.215 g (1.875 mmol, 1.3 equiv) of mesyl chloride and 0.242g (1.875 mmol, 1.3 equiv) of diisopropylethylamine. The reaction was allowed to proceed for 30 min at room temperature, solvents removed and the residue partitioned between ethyl acetate and water. Organics were dried with magnesium sulfate and purified on a silica column (1/1 ethyl acetate/hexane), yielding 0.50 g of the desired mesylate. Rf=0.5 7 (Merck Kiselgel 60, 0.25 mm, 1:1 ethyl acetate/hexane). H NMR (CDCL 3 300 MHz) 6 7.22 5H), 4.39 3H), 3.09 (dd, 1H, J=6.4, 13.2), 2.98 3H), 2.76 (dd, 1H, J=8.9, 13.2), 1.64 2H), 1.57 9H).
D.
A solution of 0.33g (0.831 mmol, 1 equiv) of the above mesylate in 3 mL DMF was cooled to 0 OC and treated with 26 mg (1.080 mmol, 1.3 equiv) of sodium hydride.
After 3h at room temperature the reaction was quenched with aqueous citric acid and purified on silica gel using 1:3 ethyl acetate/hexane The resulting product (0.18g, 72.0% yield) was then treated with 1:1 dichloromethane/ trifluoroacetic acid (5 ml) for 1/2h, WO 97/27180 PCT/US97/01610 222 resulting in 0.12g based on mesylate) of the dc ed product. IH NMR (CDCL 3 300 MHz) 5 7.23 (m, 7.04 (broad s, 1H), 3.99 1H), 2.85 2H), 2.26 (dd, 1H, J=8.1, 12.9), 1.92 (dd, 1H, J=5.0, 12.9), 1.10 2H), 0.72 2H). Low resolution MS m/e 342.2 (M+Na Example 52 1. a.LDA THF -78 °C b. acetone 2. Martin's sulfurane H
S
Bo c 3. Et 2
AICN
o o 4. TFA A solution of 1.5g (5.4 mMol) of the pyrrolidinone in mL of tetrahydrofuran was cooled to -78 °C and treated with 4.3 mL mMol) of lithiumdiisopropyl amide (2M in THF). After stirring for 0.25h, acetone (2.8g (50 mMol) was added, the reaction mixture was kept at -78 °C for 2 h and then quenched with IN hydrochloric acid. Extraction with ethyl acetate, drying over magnesium sulfate and removal of the solvent in vacuo afforded the crude product which was redissolved in 25 mL of dichloromethane and treated with 8g of Martin's sulfurane. After stirring for 12h at the mixture was participated between ethyl acetate and 1N hydrochloric acid. Drying over magnesium sulfate and removal of the solvent gave the desired alkene. 0.755 g of the crude alkene were dissolved in 15 mL of toluene and treated with 3 mL (3 mMol) of diethyl aluminumcyanide (1m in toluene) and the resulting mixture was stirred at 25 °C for 5 h.
WO 97/27180 PCT/US97/01610 223 The solvent was removed and the residue was chromatographed on silica gel (20% ethylacetatehexanes) to give the desired nitrile (0.4g) as a colorless oil. Deprotection with trifluoroacetic aciddichloromethane for 3h at 25 °C followed by chromatography on silica gel gave the desired lactam (0.22g) as a white solid. M+H: 243 Example 53 1) sodium azide 2)Pd/C I H 3)4-methoxy N NH O trityl chloride H 0
A.
A solution of 3-iodo-5-benzyl-pyrrolidinone (2.67 g, 8.87 mmol) and sodium azide (0.69 g, 10.61 mmol) in dimethylformamide (20 mL) was stirred at ambient temperature under a nitrogen atmosphere for 18h. The solvent was evaporated using a stream of nitrogen, and the residue was dissolved in ethyl acetate, washed with water and brine, and concentrated in vacuo to give a yellow solid. Chromatography on silica gel, eluting with hexane:ethyl acetate gave 1.82 g of the product as a 1:1 mixture of diastereomers which was used without separation in the next reaction. MS: ES+, 239 The chromatography also gave 0.12 g of the trans isomer as a colorless oil and 0.43 g of the cis isomer as a colorless oil which crystallized WO 97/27180 PCT/US97/01610 224 upon standing. TLC (hexane ethyl acetate Rf trans isomer =0.6 and Rf cis isomer
B.
A mixture of the above azide (0.575 g, 2.66 mmol) and 5% palladium on carbon (0.030 g) in methanol (20 mL) was stirred under 40 psi of hydrogen for 18h at ambient temperature. The mixture was filtered through a pad of Celite to remove the catalyst, followed by filtration through 5 g of silica gel, washing with chloroform:methanol The filtrate was concentrated in vacuo to give 0.46 g of the product as a mixture of diastereomers. MS: ES+, 191 and 213 (M+Na).
C.
A solution of the above amine (0.44 g, 2.3 mmol), 4anisylchlorodiphenylmethane (0.71 g, 2.3 mmol) and triethylamine (0.5 mL, 3.5 mmol) in dichloromethane mL) was stirred under a nitrogen atmosphere at ambient temperature for 18h. The solution was washed with water (2x50 mL) and brine, dried (MgSO 4 and concentrated in vacuo. The residue was purified by chromatography on silica gel, eluting with hexane:ethyl acetate then with hexane:ethyl acetate to give 0.41 g of the cis isomer as a yellow solid and 0.19 g of the trans isomer as a white solid. TLC (hexane ethyl acetate Rf cis isomer =0.5 and Rf trans isomer 0.4.
WO 97/27180 PCT/US97/01610 225 Example 54 J 2 NH
N
0 .DMFI7O 0
C
Na 2
CO
3 Iodolactam 1 (prepared as described previously in Example 7) (0.55 g, 1.8 mmol) was dissolved in DMF mL) and treated with 2-fluoroaniline (Aldrich, 0.20 g, 1.8 mmol) and solid sodium carbonate (0.39 g, 3.7 mmol). The reaction was then heated to 70 oC for 24 h before the solvent was removed in vacuo. Ethyl acetate mL) and water (20 mL) were added and the layers were partitioned. The organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo.
Flash chromatography over silica gel (1:1 hexane:ethyl acetate) gave the anilinolactam 2 as a pale yellow foam. MS 285 307 (M+Na) Example NH 0 0C Nq 2
CO
3 WO 97/27180 PCT/US97/01610 226 Using the procedure described in Example 54, the anilinolactam was prepared, purified, and isolated as a beige foam. MS 285 307 (M+Na) Example 56 NH 0 1 0
C
Na 2
CO
3 N 7NH NC H
NC
Using the procedure described in Example 54, the anilinolactam was prepared, purified, and isolated as a beige foam. MS 292 314 (M+Na) Example 57 S NH 0
TNH
O-H 0 NH 2 EtOH/A 1J~N~JNa 2
CO
3 Iodolactam 1 (prepared as described previously in Example 7) (0.77 g, 2.6 mmol) was dissolved in absolute ethanol (10 mL) and treated with 3-aminopyridine (0.26 g, 2.8 mmol) and solid sodium carbonate (0.40 g, 3.8 mmol). The reaction was then heated at reflux for 24 h before the solvent was removed in vacuo. Chloroform WO 97/27180 PCT/US97/01610 227 mL) and water (20 mL) were added and the layers were partitioned. The organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo.
Preparatory silica gel TLC (95:5 chloroform:methanol) gave the pyridylaminolactam 2 as a red oil. MS 268 290 (M+Na) Example 58 A. KCN, DMF B. H 2 Pd/C, HCI C. Ph 3 CCI, DIEA H P Ph N N Ph 1. 2.
A.
To a solution of iodolactam 1 (13.43 g, 44.6 mmol, 1 eq) in dimethylformamide (60 mL) under nitrogen was added potassium cyanide (3.49 g, 1.2 eq). After stirring at ambient temperature for 24 h, the reaction mixture was evaporated in vacuo and the residue was partitioned between ethyl acetate, saturated aqueous brine and water. The layers were separated and the aqueous layer was back-extracted twice with ethyl acetate. The combined organic layers were washed with saturated aqueous brine, dried over anhydrous magnesium sulfate, filtered and evaporated in vacuo. The residue was purified by flash silica gel chromatography eluting with hexane acetone Fractions containing the product were combined, evaporated in vacuo to provide WO 97/27180 PCT/US97/01610 228 5.89 g of cyanolactam as a mixture of diastereomers. MS (APCI): M+Na 223.
B.
A solution of cyanolactam (5.78 g, 28.9 mmol) from step A in absolute ethanol (233 mL) under Nitrogen was combined with 10 wt.% Palladium on charcoal (2.33 g) and concentrated hydrochloric acid (9.31 mL, 4 eq.).
The mixture was reduced under hydrogen gas at 50 psi for 16 h. The reaction was purged with nitrogen, filtered and evaporated in vacuo. The residue was combined with toluene 100 mL) and concentrated in vacuo to a residue to remove residual water. The azeotropic removal with toluene was repeated four times leaving a residue which was dried under high vacuum to provide the crude amine as a gum (7.18 g, 103%). MS (ESI): M+1 205.
C.
The crude amine (7.16 g, 29.8 mmol, 1 eq) from step B was combined under argon in dichloromethane (100 mL) with diisopropylethylamine (13 mL, 74.4 mmol, 2.5 eq) and triphenylmethylchoride (9.13 g, 32.7 mmol, 1.1 eq).
After stirring at ambient temperature for 16 h, the reaction mixture was treated with 5% w/v aqueous potassium carbonate and transferred to a separatory funnel. After separating the layers, the aqueous layer was back-extracted with dichloromethane and the combined organic layers were dried over anhydrous sodium sulfate and evaporated in vacuo to proved a crude mixture of diastereomers. The mixture was WO 97/27180 PCT/US97/01610 229 purified by flash silica gel chromatography eluting with ethyl acetate hexane Fractions containing the less polar diastereomer were combined and evaporated in vacuo to provide 3.52 g (26 of trityl protected amine as a crystalline solid. MS (APCI): M+Na 469.
Example 59 An alternate procedure for the synthesis of the benzyllactam:
A.
,Ph ,Ph Meo2C Ph p OHC-'NHBOC
'NHBOC
Ph Meq 2
C
A mixture of methyl 2-(triphenylphosphoranylidene)hydrocinnamate (13.20 g, 31.1 mmol, 1.15 eq) and Ntertbutoxycarobonyl-L-phenylalanal (6.76 g, 27.1 mmol, 1 eq) were combined in 200 mL chloroform and allowed to stir at ambient temperature over 64 h. The reaction was concentrated in vacuo and the residue was purified by flash silica gel chromatography eluting with 85:15 hexane ethyl acetate. Fractions containing the product were combined and evaporated in vacuo to provide the olefin as a crystalline solid (9.38 g, MS (ESI): M Na 418.
WO 97/27180 PCT/US97/01610 230
B.
Ph -Ph 1. Pd/C, H 2 h 2. TFA Ph- Ph 3. DIEA, K 2 C0 3 MeO 2
C
A solution of the olefin (9.30 g, 23.5 mmol, leq) from step A in absolute ethanol (250 mL) was combined under nitrogen with Palladium on carbon (10 wt%, 1.90 g) and reduced under a balloon of Hydrogen gas over 16 h. The reaction mixture was purged with nitrogen, diluted with dichloromethane, filtered, and evaporated in vacuo to low volume. The solution was diluted with dichloromethane and filtered through a pad of diatomaceous earth washing with dichloromethane. The filtrate was evaporated in vacuo and dried under vacuum to provide a 5:1 mixture of diastereomers of the BOCamino ester as an oil (9.68 g, 104 MS (ESI): M Na 420.
The oil was dissolved in dichloromethane (25 mL) and treated with trifluoroacetic acid (25 mL) under Argon.
After stirring for 0.5 h at ambient temperature, the reaction mixture was evaporated in vacuo. The residue was dissolved in methanol (50 mL) and treated with diisopropylethyl amine (17 mL) followed by anhydrous potassium carbonate (13.49 g, 98 mmol, 4 eq) and stirred for 16 h at ambient temperature under an Argon atmosphere. The mixture was evaporated in vacuo and the residue was partitioned between dichloromethane and water. The layers were separated and the aqueous layer WO 97/27180 PCT/US97/01610 231 was back-extracted three times with dichloromethane.
The combined organic layers-were washed with aqueous hydrochloric acid (IN) and the layers were separated.
The aqueous layer was back-extracted with dichloromethane and the combined organic layers were dried over anhydrous magnesium sulfate and evaporated in vacuo to a residue. The crude product was purified by flash silica gel chromatography eluting with a gradient of 45-60 ethyl acetate in hexane. Fractions containing the less polar diastereomer were combined and concentrated in vacuo to a solid and dried under high vacuum to provide the enantiopure lactam as a white crystalline solid (4.48 g, MS (ESI): M Na 288. H NMR (CDC13): 1.90 1H); 2.01 1H); 2.67 4H); 3.16 1H); 3.65 1H); 5.70 1H); 7.18 Example Synthesis of Compound 123
A.
N02 S O HN
K
2 C 0 3 00ON
CH
3 CN ON H
H
1 2 3 To a suspension of (2S)-(+)-glycidyl 3nitrobenzenesulfonate 1 (Aldrich, 19.47 mmol) and WO 97/27180 PCT/US97/01610 232 potassium carbonate (Baker, 38.93 mmol) in dry acetonitrile was added (S)-t-butyl decahydro-3isoquinoline carboxamide 2 (NSC Technologies, 21.41 mmol) and the reaction stirred at ambient temperature overnight. The solvent was removed in vacuo, and the residue taken up in ethyl acetate/water, the organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was purified by flash silica gel chromatography eluting with 10% diethyl' ether/dichloromethane to give 3.62 g of product 3; HPLC Rt 9.2 min TLC 1 Rf 0.26 (10% diethyl ether/dichloromethane); H NMR (CDC1 3 d 6.59 (br s, 1 3.00 1 2.97 1 2.89 (dd, 1H), 2.73 1H), 2.65 1 2.57 1 2.22 (dd, 1H), 2.08 (dd, 1H), 1.81-1.70 4 1.65-1.19 8 1.38 9 H).
B.
0 N THF, -780C to r.t.
2-pyridylmethyl lactam 1 (35 mg, 0.13 mmol) was
OH
H HbN i Phosphazene Base P4-t-Bu 3 THF, -780C to r.t.
2-pyridylmethyl lactam 1 (35 mg, 0.13 mmol) was dissolved in anhydrous THF (1 mL) and cooled to 78 OC. Phosphazene Base P 4 -t-Bu (Fluka, 1.OM in hexane, 130 uL, 0.13 mmol.) was added to give an WO 97/27180 PCT/US97/01610 233 orangish brown anion. The anion solution was stirred at -78 °C for 35 minutes and was then cannulated under nitrogen over 30 seconds into a -78 °C solution of 2 (39 mg, 0.13 mmol) in ImL of THF and was washed in with 0.5 mL of THF. The reaction was gradually warmed to room temperature over 4 hr, then stirred at room temperature for 3 days. The reaction was cooled to 78 OC, quenched with 0.5 mL of a saturated ammonium chloride solution, and concentrated in vacuo to remove the THF. The residue was partitioned between ethyl acetate and saturated bicarbonate solution and the aqueous layer was extracted with ethyl acetate. The combined organic layers were then washed with water, brine and dried (MgSO4) and filtered. Concentration in vacuo afforded 75 mg crude material which was purified via silica gel to give 18 mg(25%) of 3. Maldi MS: M H 561.5 (MW 560.79). TLC (EtOAc) Rf 0.19 (major diast.) 0.29 (minor diast.). TLC MeOH/EtOAc) Rf 0.28 (major diast.) 0.36 (minor diast.). HPLC retention times were 11.24 min. (major) 11.32 min.
(minor).
1 H NMR (CDC1 3 d 8.52 1H), 7.61 1 7.34-7.10 7H), 6.10-5.95 1H), 4.11 1H), 3.96-3.73 (m, 3H), 3.46-2.74 6H), 2.65-2.47 2H), 2.23 (m, 2H), 2.10-1.15(m, 15H), 1.37 (s,9H).
WO 97/27180 PCT/US97/01610 234 Example 61 Synthesis of Compound 72
A.
Ph Ph NaH, S (s)-epichlorohydrin I |O N, NH
N-
DMF
1 2 3-pyridylmethyl lactam 1 (85 mg, 0.32 mmol) was dissolved in DMF (1.5 mL), cooled to 0 oC, and to this solution was added sodium hydride (0.48mmol) to give a yellow anion. The reaction mixture was stirred at 0 °C for 70 minutes after which (s)-epichlorohydrin (35 ul, 0.45 mmol) was added neat. The reaction was stirred at 0 °C for 5 minutes, then warmed to room temperature and stirred for 24 hours. The reaction was cooled to 0 °C and quenched with 0.5 mL of a saturated ammonium chloride solution. The reaction was partitioned between ethyl acetate and a saturated bicarbonate solution. The aqueous layer was extracted with ethyl acetate. The combined organic layers were then washed with water, brine and dried (MgSO4) and filtered.
Concentration in vacuo afforded 49 mg of crude epoxide which was used without further purification.
WO 97/27180 WO 9727180PCTIUS97/01610 -235
B.
Ph Ph .1 OH H
N
1 2 i-PrOH0 Crude lactam epoxide 1 (49 mg) and decahydroisoquilolile 2 (91 mg, 0.38 mmol) were heated to 65-70 'C in isopropanol. After 90 hours the reaction was cooled to 25 OC and stirred for 1 hour at room temperature. The reaction was then concentrated in vacuo, and purified by silica gel chromatography, eluting with 5 MeOH :EtOAc, providing 30 mg (87% pure by HPLC) of desired product 3 as a mixture of 4 diastereomers. HPLC shows 2 split peaks 11.30 min. 11.04 min.. TLC MeOH/CH 2 Cl 2 Rf 0.27. TLC MeOH/CH 2 Cl 2 Rf 0.45.
1 H NMR (CDCl 3 d 8.45-8.35 (in, 2H), 7.48 (in, 1 H), 7.35-7.09 (mn, 6H), 6.63-5.94 (in, 1H), 3.98-3.63 (in, 3H), 3.42-2.73 (in, 5H), 2.70-2.11 (mn, 5H), 2.07-1.20(m, 1 6H) 1. 36 9H) WO 97/27180 PCT/US97/01610 236 Example 62 Synthesis of Compound 54 H2 NJ 0 2 0 ONj< 1 Phosphazene Base P4-t-Bu 3 THF, -78EC to r.t.
4-pyridylmethyl lactam 1 (33 mg, 0.12 mmol) was dissolved in anhydrous THF (1 mL) and.cooled to -78 oC. Phosphazene Base P 4 -t-Bu (Fluka, 1.0M in hexane,125 uL, 0.125 mmol) was added to give a brown anion. The anion solution was stirred at -78 °C for minutes and was then cannulated under nitrogen over seconds into a -78 oC solution of 2 (39 mg, 0.13 mmol) in ImL of THF and was washed in with 0.5 mL of THF.
The reaction was gradually warmed to room temperature over 4 hr, then stirred at room temperature for 3 days.
The reaction was cooled to -78 OC, quenched with 0.5 mL of a saturated ammonium chloride solution, and concentrated in vacuo to remove the THF. The residue was then partitioned between ethyl acetate and saturated bicarbonate solution. The aqueous layer was extracted with ethyl acetate and the combined organic layers were then washed with water, brine and dried (MgSO4) and filtered. Concentration in vacuo afforded 83 mg crude material which was purified via silica gel WO 97/27180 PCT/US97/01610 237 to give 11 mg(16%) of 3. Maldi MS: M H 560.4. (MW 560.79). TLC (EtOAc) Rf 0.08 (major diast.) 0.16 (minor diast.). TLC MeOH/EtOAc) Rf 0.18 (major diast.) 0.26 (minor diast.). HPLC retention time was 11.05 min.
H NMR (CDC1 3 d 8.50 2H), 7.35-7.02 7H), 5.89 1H), 4.05-3.78 3H), 3.37-2.69 5H), 2.62- 2.45 4H), 2.26 2H), 2.08-1.16(m, 15H), 1.38 (s,9H).
Example 63 Synthesis of Compound 130 Ph Ph NH 2) OH H O O HN 0 N-H 1 2 NH
H
In an oven-dried 25 mL round-bottomed flask, alkyne lactam 1 (54.6 mg, 0.682 mmol) was dissolved in 5 mL of DMF. Sodium hydride (34.4 mg of a 60% dispersion in mineral oil, 0.860 mmol) was added with cooling using an ice-bath. Gas evolution was observed. Epichlorohydrin (60 uL, 0.765 mmol) was added. The mixture was stirred overnight at room temperature, then the decahydroisoquinoline amide (182 mg, 0.770 mmol) was added. The mixture was heated to 80 °C overnight.
The mixture was cooled, poored into water, and WO 97/27180 PCT/US97/01610 238 extracted with CH 2 Cl 2 The organic extract was washed several times with water, dried (MgSO 4 and evaporated in vacuo to afford a yellow residue that was purified by preparative HPLC to afford the a diastereomeric mixture of alkyne DHIQ lactam 2 (120 mg, 34%) as a light yellow oil. HPLC: retention times of 13.57, 13.67, 13.87 minutes in a 5:1:1 ratio respectively. H *NMR d 1.3-1.4 three singlets in a 2:1:1 ratio; 1.4- 2.7 (several overlapping multiplets, 2.8-2.95 (multiplet), 3.0-3.7 (multiplet), 3.8-4.1 (multiplet), 5.95-6.05 (multiplet), 6.18, 6.32, 6.4 (broad singlets in a 1:1:1:1 ratio), 6.2-6.3 (doublet of doublets), 7.15-7.35 (multiplet). MALDI-MS: peak at 506.3 (M Example 64 Synthesis of Compound 124
SOH
NjNH 02 NJ N 1 Phosphazene Base P 4 -t-Bu 3 THF, -78°C to r.t.
The lactam 1 (0.13 mmol) was dissolved in dry THF at -78 °C and to this solution was added Phosphazene Base
P
4 -t-Bu (Fluka, 1.0M in hexane, 0.14 mmol). After stirring 15 minutes the anion solution was added via cannula to a solution of epoxide 2 (0.13 mmol) WO 97/27180 PCT/US97/01610 239 dissolved in dry THF at -78 oC and the reaction was allowed to warm to room temperature and stir overnight.
The reaction was then diluted with water and extracted with ethyl acetate. The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was taken up in dichloromethane and filtered through a plug of silica gel eluting with 8% MeOH in dichloromethane.
Product containing fractions were concentrated in vacuo and the resultant residue further purified by preparative HPLC (column: Delta-Pak C18 15mm 100A 19x300 mm. Gradient: 20% to 100% acetonitrile in water with 0.1% TFA. Flow rate: 20 ml/min.
Detection: 214 nm) to yield 3 mg of product 3 as a mixture of diastereomers; TLC Rf 0.44 (8% MeOH/CH 2 C12); HPLC Rt 14.8, 14.9 min MALDI- TOF MS m/z 561 1 H NMR (CDC13) d 7.35-7.10 7 6.73 1 6.58 2 5.82 (br s, 1 H), 4.12-3.85 4 3.51 1H), 3.30 1H), 2.92 1 3.63-2.20 4 2.05-1.12 18 1.38 9 H).
WO 97/27180 PCT/US97/01610 240 Example Synthesis of Comoound 127
OH
NC" INH HN NC
SH
0 2 ON 1 Phosphazene Base P 4 -t-Bu 3 THF, -780C to r.t.
Cyanomethyl lactam 1 (82 mg, 0.38 mmol) was dissolved in anhydrous THF (2 mL) and cooled to -78 OC.
Phosphazene Base P 4 -t-Bu (Fluka, 1.0M in hexane, 380 uL, 0.38 mmol) was added to give a yellow anion. The anion solution was stirred at -78 °C for 35 minutes and was then cannulated under nitrogen over 30 seconds into a -78 OC solution of 2 (112 mg, 0.38 mmol) in 2mL of THF and was washed in with 0.5 mL of THF. The reaction was gradually warmed to room temperature over 4 hr, then stirred at room temperature for 3 days. The reaction was cooled to -78 OC, quenched with 0.5 mL of a saturated ammonium chloride solution, and concentrated in vacuo to remove the THF. The residue was then partitioned between ethyl acetate and saturated bicarbonate solution and the aqueous layer was extracted with ethyl acetate. The combined organic layers were then washed with water, brine and dried (MgSO4) and filtered. Concentration in vacuo afforded 375 mg crude material which was purified via silica WO 97/27180 PCT/US97/01610 241 gel (8 2, ethyl acetate: CH 2 Cl 2 to give 118 mg(61%) of 3 that was 80% pure. 58 mg was purified via prep.
HPLC to give 10 mg of pure material as a 2: 1 mixture of diastereomers. HPLC retention times were 12.73 min.
12.86 min. Maldi MS: M H 510.47 (MW 508.71). TLC EtOAc) Rf 0.37 0.31.
H NMR (CDC1 3 d 7.38-7.13 5H), 6.09-5.82 (br s, 1H), 4.29-3.96 3H), 3.84 1H), 3.49-2.91 (m, 2.77-2.18 9H), 2.10-1.20(m, 11H), 1.39 (s,9H).
Example 66 Synthesis of Compound 131 NNH O NN N N 0 2H
O
1 Phosphazene Base P 4 -t-Bu 3 H THF, -78C to r.t.
The lactam 1 (0.061 mmol) was dissolved in dry THF at -78 OC and to this solution was added Phosphazene Base
P
4 -t-Bu (Fluka, 1.OM in hexane, 0.067 mmol). After stirring 15 minutes the anion solution was added via cannula to a solution of epoxide 2 (0.061 mmol) dissolved in dry THF at -78 °C and the reaction was allowed to warm to room temperature and stir overnight.
The reaction was then diluted with water and extracted with ethyl acetate. The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, WO 97/27180 PCT/US97/01610 242 followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was purified by flash silica gel chromatography eluting with 3% MeOH in dichloromethane to give 2.1 mg of product 3 as a 1:1 mixture of diastereomers; TLC Rf 0.14 (2:1 ethyl acetate/hexanes); HPLC Rt 13.6, 13.8 min MALDI-TOF MS m/z 580 (M 1H NMR (CDCl 3 d 7.32-7.08 5 5.86 (br s, 1 4.08-3.73 4 3.65- 3.14 4H), 3.00-2.49 8H), 2.41-0.92 13 H), 2.27 1.5 2.22 1.5 2.16 1.5 2.11 1.5 1.46 9 H).
Example 67 Synthesis of Compound 126 2 1 Phosphazene Base P 4 -t-Bu 3 H THF, -78 oC to r.t.
The lactam 1 (0.20 mmol) was dissolved in dry THF at -78 °C and to this solution was added Phosphazene Base
P
4 -t-Bu (Fluka, 1.OM in hexane, 0.21 mmol). After stirring 15 minutes the anion solution was added via cannula to a solution of epoxide 2 (0.20 mmol) dissolved in dry THF at -78 °C and the reaction was allowed to warm to room temperature and stir overnight.
The reaction was then diluted with water and extracted WO 97/27180 PCT/US97/01610 243 with ethyl acetate. The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was taken up in dichloromethane and filtered through a plug of silica gel eluting with 3% MeOH in dichloromethane.
Product containing fractions were concentrated in vacuo and the resultant residue further purified by preparative HPLC (column: Delta-Pak C 18 15mm 100A 19x300 mm. Gradient: 20% to 100% acetonitrile in water with 0.1% TFA. Flow rate: 20 ml/min.
Detection: 214 nm) to yield 2.5 mg of product 3 as a mixture of diastereomers; TLC Rf 0.21 (3% MeOH/CH 2 Cl 2 HPLC Rt 14.8 min MALDI-TOF MS m/z 588 Example 68 Synthesis of Compound 132 Ph O Ph S 1) H.
OH
S1)N 0
OH
H 2) H S2 0 N-H In an oven-dried 25 mL round-bottomed flask, isoxazole lactam 1 (54.6 mg, 0.201 mmol) was dissolved in 3 mL of THF. (S)-Epichlorohydrin (20 uL, 0.255 mmol) was added. P-4-tBu phosphazene base (210 uL, 0.210 mmol) WO 97/27180 PCT/US97/01610 244 was added dropwise via syringe initially producing a dark orange-brown color that faded. The mixture was stirred for 30 minutes at room temperature and the mixture was poured into water and extracted with
CH
2 Cl 2 The organic extract was dried (Na 2
SO
4 and evaporated in vacuo.. The residue was dissolved in anhydrous CH 3 CN and the decahydroisoquinoline amide (54.4 mg, 0.230 mmol) was added. The mixture was refluxed overnight. The solvent was evaporated and the residue was purified by preparative HPLC to afford the isoxazole DHIQ lactam 2 (38 mg, 34%) as a light yellow oil. HPLC: retention times of 12.28, 12.86, 13.68 minutes at 93% purity. H NMR d 1.3-1.4 three singlets in a 4:4:1 ratio; 1.4-2.7 (several overlapping multiplets, 1.4-2.3 (several overlapping multiplets), 2.45-3.35 (several overlapping multiplets), 3.35-4.1 (several multiplets), 4.3-4.4 (doublet), 5.8 (multiplet), 5.9, 6.0, and 6.3 (three broad singlets in a ratio of 4:4:1 ratio), 7.1-7.2 (multiplet), 7.2-7.4 (multiplet). MALDI-MS: calc'd: 564.9; found 565.5 (M
H
WO 97/27180 PCT/US97/01610 245 Example 69 Synthesis of Compound 125
OH
2 0 ONN N0 1 Phosphazene Base P 4 -t-Bu 3 H THF, -780C to r.t.
The lactam 1 (0.12 mmol) was dissolved in dry THF at -78 OC and to this solution was added Phosphazene Base
P
4 -t-Bu (Fluka, 1.OM in hexane, 0.13 mmol). After stirring 15 minutes the anion solution was added via cannula to a solution of epoxide 2 (0.12 mmol) dissolved in dry THF at -78 oC and the reaction was allowed to warm to room temperature and stir overnight.
The reaction was then diluted with water and extracted with ethyl acetate. The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was taken up in dichloromethane and filtered through a plug of silica gel eluting with 3% MeOH in dichloromethane.
Product containing fractions were concentrated in vacuo and the resultant residue further purified by preparative HPLC (column: Delta-Pak C 18 15mm 100A 19x300 mm. Gradient: 20% to 100% acetonitrile in water with 0.1% TFA. Flow rate: 20 ml/min.
Detection: 214 nm) to yield 1.5 mg of product 3 as a WO 97/27180 WO 9727180PCT/US97/01610 -246 single diastereoner; TLC Rf =0.27 MeOH/CH 2 Cl 2 HPLC Rt 14.7 min MALDI-TOF MS m/z 576 1H NMR (CDCl 3 d 7.40-7.15 (in, 7 6.70 (in, 21 6.55 2 5.80 (br s, 21 4.28 (mn, 1 H), 4.05-3.90 (in, 2 3.70-3.38 (mn, 2H), 3.20 (mn, 1H), 3.00-2.75 (in, 2 2.70 3 2.55 (in, 2H), 2.30 (in, 2H), 2.20-0.80 (mn, 14 1.35 9 H).
Sunthesis of Conound 128 A
NH
EtOOC 0, Phosphazene Base THF,-780C The lactarn 1 (90 mng, 0.28 iniol) was dissolved in THF (3 niL) and cooled to -78 OC. This was followed by the addition of the phosphazene base (Fluka; 1M in hexane, 0.28 niL, 0.28 mmiol) After stirring at -78 OC for one hour the epoxide was added as a solution in 1 niL THF.
The reaction was then warmed to 25 OC and stirred for an additional 3 hours. The reaction was then quenched WO 97/27180 PCTIUS97/01610 247 by the addition of water and extracted by ethyl acetate. The organic portion was then dried over MgSO 4 filtered and concentrated in vacuo. The crude oil was purified by silica gel chromatography, eluting with 1:1, ethyl acetate:hexanes, this provided the two major products (HPLC.indicated two components for each isolate).
B.
OH f LiOH OH Et 0 N H 0 N H 1 2 To the elaborated lactam 1 (40 mg) in 2:1, THF:H 2 0 mL) was added LiOH (2 The reaction was then stirred at 40 oC for 16 hours. TLC indicated the formation of a new component. The reaction was diluted by ethyl acetate, after which the organic portion was separated, dried over MgSO 4 filtered and concentrated in vacuo. to yield product 2 as a mixture of diastereomers.
H NMR (CDC1 3 d 7.10-7.50 10 5.90-6.15 (m, 1H), 3.90-4.40 2H), 3.20-3.70 3H), 2.80-3.10 2H), 2.60-2.70 2H), 2.20-2.60 3H), 1.60- 2.10 9H), 1.40 15H), 1.20-1.40 8H).
WO 97/27180 PCT/US97/01610 248 Example 71 Synthesis of Compound 259 H o
OH
2
O
1 Phosphazene Base P 4 -t-Bu 3
H
THF, -780C to r.t.
The lactam 1 (0.11 mmol) was dissolved in dry THF at -78 OC and to this solution was added Phosphazene Base
P
4 -t-Bu (Fluka, 1.OM in hexane, 0.12 mmol). After stirring 15 minutes the anion solution was added via cannula to a solution of epoxide 2 (0.11 mmol) dissolved in dry THF at -78 °C and the reaction was allowed to warm to room temperature and stir overnight.
The reaction was then diluted with water and extracted with ethyl acetate. The organic layer was washed sequentially with saturated aqueous NaHCO3 and brine, followed by drying (MgSO4), filtration and concentration in vacuo. The crude residue was taken up in dichloromethane and filtered through a plug of silica gel eluting with 5% MeOH in dichloromethane.
Product containing fractions were concentrated in vacuo and the resultant residue further purified by preparative HPLC (column: Delta-Pak C 18 15mm 100A 19x300 mm. Gradient: 20% to 100% acetonitrile in water with 0.1% TFA. Flow rate: 20 ml/min.
WO 97/27180 PCT/US97/01610 249 Detection: 214 nm) to yield 12 mg of product 3; TLC Rf 0.50 MeOH/CH 2 Cl 2 HPLC Rt 12.8 min (100%); MALDI-TOF MS m/z 541 (M 1H NMR (CDC1 3 d 7.35-7.16 5 5.86 (br s, 1 4.08-3.76 4 3.49- 3.22 4 2.89 (br s, 1 2.50 2 2.25 (br s, 1H), 2.14-1.11 22 1.38 9 H).
Example 72 Synthesis of Compound 260 PPh PhPh Ph Ph S H
OH"H
H
0 N-H NH FH In an oven-dried 25 mL round-bottomed flask, triazole lactam 1 (124 mg, 0.358 mmol) was dissolved in 5 mL of THF. (S)-Epichlorohydrin (50 uL, 0.639 mmol) was added. P-4-tBu phosphazene base (370 uL of 1.OM solution in hexane, 0.370 mmol) was added dropwise via syringe initially producing a dark orange-brown color that faded. The mixture was stirred for 30 minutes at room temperature and the decahydroisoquinoline amide (124 mg, 0.525 mmol) was added. The mixture was refluxed overnight. The solvent was evaporated and the residue was purified by preparative HPLC to afford the triazole DHIQ lactam (189.1 mg, HPLC: retention times of 12.94, 14.22 minutes at 99% purity.
WO 97/27180 PCT/US97/01610 250 1 H-NMR d1.3-1.4 two singlets in a 1:1 ratio; 1.4-3.1 (several overlapping multiplets, 1.4-2.3 (several overlapping multiplets), 2.45-3.35 (several overlapping multiplets), 3.2-4.2 (several multiplets), 5.4-5.6 (multiplet), 6.1 and 6.45 (two broad singlets in a 1:1 ratio), 5.9, 6.0, and 6.3 (three broad singlets in a ratio of 4:4:1 ratio), 7.1 (doublet), 7.2-7.5 (multiplet). MALDI-MS: calc'd (-DHIQ): 401.2; found 403.6 (M DHIQ 2H+).
Example 73 Synthesis of Compound 129 S H H Ph H OH tributyl tin azide ,N OH heat 0 NH O ONH 2 In a heavy-walled screw-top test tube, the alkyne lactam 1 (83 mg, 0.164 mmol) was dissolved in 5 mL of xylene. Tributyltin azide (200 mg, 0.602 mmol) was added, the tube was sealed and heated to 2050 C overnight. The dark brown solution was cooled and directly chromatographed using a gradient from CH 2 Cl 2 to 50% EtOAc/MeOH to afford the triazole product 2 (14 mg, as a light yellow oil. HPLC: retention times of 12.01, 12.44, 13.01, 13.22 minutes in an 8:4:1:1 ratio at 99% purity. MALDI-MS: calc'd (-DHIQ): 550.4; found 552.9 (M 2H+).
WO 97/27180 PCT/US97/01610 251 Exampne 74 Synthesis of Compound 227 phosphazene base, THF, -780C H
H
1 2 o 2 isopropanolconc. HCI rt To a cooled solution (-78 OC) of lactam 1 (0.10g, 0.46mmol) in anhydrous THF (l.OmL) was added phosphazene base P4 t-butyl solution (1.OM in hexanes, 0.46mL,0.46mmol) with stirring. After a 15 min.
stirring period, epoxide 2 (0.173g,0.46mmol) was added in one portion and the reaction was allowed to slowly warm to rt. After 0.5h at rt, 1.OM HC1 (10.OmL) was added and the solution was diluted with ethyl acetate The ethyl acetate was washed with sat. NaHCO 3 (1 x 10mL), brine (1 x 10mL) dried (MgS04), filtered, and evaporated to give a brown foam. The crude acetonide (0.270g, 0.46mmol) was dissolved in isopropanol (10mL) and treated with conc. HC1 at rt. After 2.0h., the solution was adjusted to pH 11 with 3.ON NaOH and extracted with ethyl acetate (3 The ethyl acetate was dried(MgSO 4 and evaporated to give the crude product which was purified by column chromatography: methylene chloride/methanol (98/2) to give the product as an off white solid (0.090g, MS: crude acetonide: M+Na 617; WO 97/27180 WO 9727180PCTIUS97/0 1610 -252 product: M+Na 571 HNNR (CDC1 3 0-90(m, 6H); 1.15(m, 1H); 1.40(m, 1H); 1.50-1.80(m, 2H); 1.90(m,1H); 2.18(m, 2.25H); 2.30-2.50(m, 1H); 2.60(m, 0.75H); 2.80-3.10(mn, 4H); 3.30(m, 2H); 3.60(m, 1.25H); 3.80(m, 1.75H); 3.95(m, 1H); 4.25(m, 1H); 4.40(m, 0.75H); 5.00(m, 0.25H); S.25(m, 1H); 5.95(d, 0.25H); 6.10(d, 0.75H); 7.00-7.40(m, 14H) Exjample Synthesis of Compound 232 q 1) NaH, DMF, 80 0
C
oconc. HCI OH H PH H3CO 1-1cc 0 S 2 Prepared using the procedure outlined in Example 24.
The acetonide was purified by column chromatography: 60/40 hexane/ethyl acetate. MS: N+NA 647. The product was purified by column chromatography: 98/2
CH
2 Cl 2 /MeOH. MS: M+H 585 1H NMR (ODC1 3 1.70(m, 2H); 1s 1.80(m, 1H); 1.90(m, 1H); 2.10(m, 1H); 2.40-3.10(m, 3.60(s, 3H); 3.75(m, 1H); 3.90(m, 1H); iH); 4.30(m, 3H); 5.30(m, 1H); 6.10(d, 1H); 7.00- 7.40(m, 14H).
WO 97/27180 WO 9727180PCTIUS97/01610 253 xaple 76 Synthesis of Compound-231 1) NaIL DMIT, S0 0
C
conc. HCI Prepared using the procedure outlined in Example 24.
The acetonide was purified by column chromatography: 98/2 CH 2 Cl 2 /MeOH. MS: M+H =642. The product was purified by column chromatography: 96/4 CH 2 Cl 2 /MeOH.
MS: M+H 602 1H NMR. (CDCl 3 1.50-2.50(m, 6H); 2.50- 3.40(m, 6H); 3.50-4.40(m, 7H); 5.25(m, 1H); 5.95(m, 1H); 7.00-7.60(m, 18H).
-0 Example 77 Synthesis of Compound 216 91) NaH, DMF, 80 0
C
conc. HCI Z I 7( )4H 1 4OHH
OH
OJ-p3 WO 97/27180 WO 9727180PCTIUS97/01610 -254 Prepared using the procedure outlined in Example 24.
The acetonide was purified by column chromatography: 50/50 hexane/ethyl acetate. MS: M+NA =645. The product was purified by column chromatography: 96/4
CH
2 Cl 2 /MeOH. MS: M+NA 605 1H NM'R ('CDCl 3 1.10-1.40(m, 2H); 1.70(m, 2H); 1.80-2..10(m, 4H); 2.35(m, 2.50(m, 2.65(m,0.5H); 2.80-3.10(m, 4H); 3.20(m, 2H); 3.30-3.55(m, 3H1); 3.70(m, lIH); 3.80-4.00(m, 4H); 4.25(m, 1H1); 4.37(m, 1H1); 5.27(m, 1H1); 6.15(d, 1H1); 7.10-7.40(m, 14H).
Exam le 78 Synthesis of Compound 221 1) NaDMF, 90 *C conc. HCI OH HOH NNH
NN
0 00 00 0YN8 3 8 2 Prepared using the procedure outlined in Example 24.
The acetonide was not purified by column chromatography. MS: (crude) M+H 644. The product was purified by column chromatography: 96/4 CH 2 Cl 2 /MeOH.
MS: M+H 604 1 H NMP. (CDCl 3 1.40-2.20(m, 6H); 2.30(m, 1H); 2.50-3.40(m, 9H); 3.75(m, 2H); 4.00(m,lH); 4.25(m, 111); 5.30(ma, 1H); 6.35(d, 0.5H); 6.50(d, 0.5H1); 7.00- 7.40(m, 14H); 7.50(m, 2H); 8.50(m, 2H1).
WO 97127180 WO 9727180PCTIUS97/01610 255 Example 79 Synthesis of Compound 223 1) NaHDMF.S 800C
N-
0 conc. HCI
NH~N-
0 OH H OH 21 N.8o 03 2 Prepared using the procedure outlined in Example 24.
The acetonide was not purified by column chromatography. MS: (crude) M+NA 670. The product was purified by column chromatography: 97/3 CH 2 01 2 /MeOH.
MAS: M+NA 630 1 1H NM?. (CDC1 3 1.40(mn, 1H); 1.30-1.80(m, 2H); 1.95(m, 1H); 2.10(m, 1H); 2.25(m, 2H); 2.30- 3.40(m, 7H); 3.60-3.80(m, 2H); 3.85(m,lH); 4.00(m, 1H); 4.25(m, 1H); 4.45(m,lH); 5.30(m, 1H); 5.80(m,lH); 6.15(d, IH); 7.10-7.40(m, 14H).
WO 97/27180 WO 9727180PCTIUS97/01610 256 Example S=thesis of Compound 230 9 1) NaHDME.30 0
C
conc. HCI Prepared using the procedure outlined in Examiple 24.
The acetonide was not purified by column chromatography. MS: (crude) M±NA 746. The product was purified by column chromatography: 97/3 CH 2 Cl 2 /MeOH.
MS: M+NA 706.
Example 81 Synthesis of Compound 224 1) NaKl DMP, 80 0
C
conc. HCI WO 97/27180 WO 9727180PCTIUS97/0 1610 -257 Prepared using the procedure outlined in Example 24.
The acetonide was not purified by column chromatography. MS: (crude) M+NA 673. The producz was purified by column chromatography: 96/4 CH 2 Cl 2 /MeOH.
MS: M+NA 633 1H NMR (CDCl 3 0.090'-1.30(m, 4H); 1.40- 1.80(m, 4H); 1.90-2.35(m, 3H); 2.45 (in, 1H); 2.65(m, 1H); 2.70-3.10(m, 6H); 3.25(m, 3H); 3.60-4.00(m, 6H); 4.25(m, 1H); 4.35(m, 4.75(m, 0.SH); 5.25(m, 1H); 6.20(m, 7.10-7.40(m, 14H).
Example 82 Synthesis of Compound 225 phosphazene base, THF, -78'C V j isopropanokconc. HCI rt Prepared using the procedure outlined in Example 74.
The acetonide was not purified by column chromatography. MS: (crude) 2M-sNA =1179. The product 1s was purified by column chromatography: 80/20 ethyl acetate/hexane. MS: M+H 539 1 H NMR (ODC1 3 0. 55 (in, 1H)f; 0.659m, 0.95(m, 1H); 1.05(m, 1H); 1.75(mn, 1.95(m, 2.20 (dd, 2.65(dd, 1H); 2.70- 3.10(m, 6H); 3.20(d, 1H); 3.65(dd, 1H); 3.95mn, 2H); 4.25(t, 1H); 5.25(mn, 1H); 5.95(d, 1H); 7.10-7.40(m, 14H).
WO 97/27180 WO 9727180PCTIUS97/01610 -258 Exaple83 Synthesis of Comipound 226 c~1) NaN, DMF, 90 0
C
conc. HCI
O
0 0 3 2 8 Prepared using the procedure outlined in Example 24.
The acetonide was purified by column chromatography: 60 40 hexane/ethyl acetate. MS: M+NA 666. The product was purified by column chromatography: 40/60 hexane/ethyl acetate. MS: !A+H 604 1H MR 1. (in, 0.5H); 1.70m, 0.5H); 1.95(m, 1H); 2.50 (in, 1H); 2.70-3.10(m, 7.5H); 3.15(dd, 1H); 3.30(m, 1H); 3.40(m, 1H); 3.75(m, 1H); 3.80-4.10(m, 2H); 4.25(m, 4.45(mI 0.5H); 5.25(m, 1H); 6.15(m, 1H); 6.45(d, 1H); 6.55(q, iN); 6.70(q, iH) 7.10--7.40(m, 16H).
WO 97/27180 WO 9727180PCTfUS97/01610 259- Examrle 8 Synthesis of Compound 229 2 OH H9OH NC1.) phosphazene base, THF, -78 0 C
NC
H 0 0 isomer 1 1 822 ,,OH H9 isopropanollconc. HCI rtN isomer 2 Prepared using the procedure outlined in Example 74 The acetonide was purified by column chromatography and the diasteriomers were isolated separately. MS: (isomer 1) 14+NA 642; (isomer 2) M+NA 642. The individual diastereomers were deprotected and purified by column chromatography: 98/2 CH 2 Cl 2 /MeOH to give isomer 1 and isomer 2. MS: (isomer 1) M±NA 602; (isomer 2) M+NA 602. 1HNMR (CDCl 3 isomer 1: 1.05 1H); 1.35(s, 3H); 1.45 3H); 1.75(m, 1H); 1.90-2.20(m, 3H); 2.65(m, 1H); 2.70-3.10(m, 8H); 3.70(m, 1H); 3.95(m, 2H); 4.20(m, 1KH); 4.35(m, 1H); 5.25(m, 1H); 6.05(d, 1H); 7.10-7.40(m, 14H). 1H NMR (ODC1 3 isomer 2: 1.10 1H); 1.40(s, 3H); 1.50 3H); 1.75(m, 1H); 1.95(m, 1H); 2.15(m, 1H); 2.50(m, 2H); 2.80-3.10(m, 6H); 3.35(m, 2K); 3.65(m, 1H); 3.80(m, 1H); 4.00(m, 2H); 4.25(m, 1H); 5.25(m, 1H); 5.95(d, 1H); 7.10- 7.40(m, 14H).
WO 97/27180 WO 9727180PCTIUS97/01610 260 Example synthesis of Compound-261 1phosphazene base, THF, -78 0 COH H N2.) isomer 1 H3632 3 is o p ro p a n o llc o n c H C I rt 36i o e 2 Prepared using the procedure outlined in Example 74.
The acetonide was purified by column chromatography: 60/40 hexane/ethyl acetate and the diasteriomers were isolated separately. MS: (isomer 1) M+H 658; (isomer 2) M+H 658. The individual diastereomers were deprotected and purified by column chromatography: 40/60 hexane/ethyl acetate to give isomer 1 and isomer 2. MAS: (isomer 1) M+H 618; (isomer 2) M+NA 640. 1H NMRP (ODC1 3 isomer 1: 1.75(m, 1H); 1.90-2.20(m, 3H); 2.70(s, 3H); 2.75-3.15(m, 6H); 3.75(m, 1H); 4.00(m, 3H); 4.25(m, 1H); 4.65(m, iH); 5.25(m, 1H); 6.05(d, 1H); 6.55(dd, 2H); 6.70(m, 1H); 7.00-7.40(m, 16H). 1H NM'R (ODC1 3 isomer 2: 1.70(m, 2H); 1..95(m, 1H); 2.25(m, 1H); 2.55(m, 1H); 2.70(s, 3H); 2.80-3.10(m, 8H); 3.35(dd, 1H); 3.40(dd, 1H); 3.75(m, 1H); 3.80(m, 1H); 4.05(m, iH); 4.25(m, 1H); 4.55(t, 1H); 5.30(m, 1H); 6.05(d, 1H); 6.70(m, 2H); 7.10-7.40(m, 17H).
WO 97/27180 WO 9727180PCTIUS97/01610 261 Exarmle 8 Synthe~sis of Compound 228
H
.N OH H OH 0 0Q .NN OH H OH 20% Pd/C isomer 1
NN
se0ae 8 H 9 sprtddiastriomers N' H H O isomer 2 N The benzyl triazole from Example 80 was purified (and diastereomers isolated) by column chromatography: 97/3
CH
2 Cl 2 /IMeOH. MS: M+NA 706. The individual benzyl protected diastereomers were dissolved in MeOH and combined with 20% Pd/C Each solution was hydrogenated under pressure(S0 psi) at rt for 5 days and the resulting crude product was purified by column chromatography 96/4 CH 2 Cl 2 /bMeOH to give isomers 1 and 2. MS: (isomer 1) M+H 594; (isomer 2) M+NA =616. 1H NI4R (CDCl 3 isomer 1: 1.60(m, 1H); 1.80(m, 2H); 2.40(s, 1H); 2.60-3.15(m, 10H); 3.65(m, 1H); 3.80(m, 1H); 4.00(m, 1H); 4.20(m, 1H); S.2S(m, 1H); 6.90(m, 1H); 7.00--7.40(m, 14H). NMR (CDCl 3 isomer 2: 1.30(m, 1H); l.75(m, 1H); 1.95(m, 2H); 2.35(m, IN); 2.50(m, 1H); 2.80-3.10(m, 8H); 3.25(d, 1H); 3.65Cm, 1H-); 3.80(m, 1H); 4.05(m, 1H); 4.30(m, 1H); 4.50(m, 1H); 5.25(m, IN); 6.75(m, 1H); 7.10-7.40(m, 14H).
WO 97/27180 WO 9727180PCT/US97/0 1610 262 Examole 87 Synthesis of Compound 219
N
separated diastereomers isomers 1 ,2,38 Isomer 1: Prepared using the procedure outlined in Example 24. The acetonide was purified by column chromatography 30/70 hexane/ethyl acetate MS: M'+NA 645. The product was purified by column chromatography: 30/70 hexane/ethyl acetate MAS: M+NA 605 1 NM'R (CDCl 3 1.45(m, 1H); 1.70(m, 1H); 1.80-2.05(m, 4H); 2.25 1H); 2.35(q,1lH); 2.65(m, 1H); 2.75-3.10(m, 8H); 3.60(m, 3H); 3.75(m,lH); 3.85(m, 1H); 3.95(m, 2H); 4.25(m, 1Hi); 5.25(m, iH); 6.05(m, 1H); 7.10-7.40(m, 14H).
Isomers 2,3: (Chiral center within TEF ring has opposite configuration to that of isomer 1 above) Prepared is using the procedure outlined in Example 74. The acetonide was purified (diastereomers isolated) by column chromatography 30/70 hexane/ethyl acetate MS: M+NA 645. The individual diastereomers were purified by column chromatography: 30/70 hexane/ethyl acetate MAS: (isomer 2) M+NA 605 (isomer 3) M+NA 605 1 H NMR (CDCl 3 (isomer 2) 1.45(m, 2H1); 1.90(m, 2H1); 2.10(m, 1H1); 2.30 (mn, 1H); 2.35(m, 1H); 2.45(m, 1H); 2.75- 3.10Cm, 6H1); 3.25(m, 2H1); 3.65(m, 3H1); 3.75(m,3H); 3.95(m, 2H); 4.25(m, 2H1); 5.25(m, 1H); 6.10(m, 1H); WO 97127180 WO 9727180PCT/US97/01610 263 7.10-7.40 14H). H NNR (CDC1 3 (isomer 3) 1.15(mn, 1H); 1.80(m, 1H); 1.95(m, 2H); 2.10(m, 2.25 (mn, 1H); 2.40(m, 1H); 2.60(m, 1H); 2.75-3.10(m, 8H); 3.40(m, 1H); 3.60-4.00(m, 6H); 4.25(m, 1H); 5.25(mn, 1H); 6.05(m, 1H); 7.10-7.40(m, 14H).
Examle 88 Synthesis of Compound 233 OH H OH phasphazene base, THF, -71C N ~N 9NH -10isomer 1 N 0 P) ciisomer 2 Prepared using the procedure outlined in Example 74.
The acetonide was purified (diastereomers isolated) by column chromatography 45/55 hexane/ethyl acetate MS: M'+NA =692. The individual diastereomers were purified by column chromatography: 35/65 hexane/ethyl acetate MS: (isomer 1) M+H 630 (isomer 2) M+H 630 1H NMR (CDCl 3 (isomer 1) 1.75(m, 1H); 1.95(m, 1H); 2.10(mn, 2H); 2.75-3.10(m, 8H); 3.15(d, 2H); 3.30(in, 2H); 3.80(m,2H); 4.00(m, 2H); 4.25(m, iH); 5.25(m, 1H); 6.00(m, iH); 6.20(d, 1H); 6.60(t, iH); 6.95-7.40(mn, 18H). 1HNMR (ODCd 3 (isomer 2) 1.75(m, 1H); 1.95(m, 2H); 2.15(m, 1H); 2.55 (mn, iH); 2.75-3.10(m, BH); 3.20- WO 97/27180 WO 9727180PCTIUS97/01610 264 3.50(m, 4H); 3.75 (mn, 1H); 3.85(in,1H)4.OO(n, 1H); 6.30(mn, 1H); 6.56(t, 1H);7.1-7.40(m, 18
H).
Exaiple 89 Synthesis of Comp~ound 234
NH
9 phosphazene base, THF. -781C 2.) isopropanollconc. HCI rt Prepared using the procedure outlined in Example 74.
The acetonide was purified by column chromatography: 97/3 0H 2 01 2 /MeOH. MS: M±I- 633. The product was not purified. MS: 593.
WO 97/27180 PCT/US97/01610 265 Example Synthesis of Compound 235
A.
HCO 1) Na DMF.
0 C SOD S 2) conc. HCI OH O OH H N NH 3) methylN 1 chloroformate H 0 H O R, 3 2 A mixture of the trans isomer of the lactam above (0.125 g, 0.27 mmol) and 60% sodium hydride (0.010 g, 0.25 mmol) in dimethylformamide (4 mL) was stirred under a nitrogen atmosphere for 30 min. The epoxide (0.113 g, 0.30 mmol) was added and the mixture was heated at 60 oC for 4h. The mixture was re-charged with 60% sodium hydride (0.015 g, 0.37 mmol) heated at oC for an additional 1.5h, and stirred at ambient temperature for 18h. The mixture was diluted with dichloromethane, washed with brine, dried (MgSO 4 and concentrated in vacuo. The residue was purified by chromatography on silica gel, eluting with hexane:ethyl acetate then with hexane:ethyl acetate to give 0.llg of product as a brown oil. MS: AP+, 862 (M+Na) and AP-, 874 (M+Cl).
B.
A solution of the acetonide (0.llg, 0.13 mmol) in 2propanol (7 mL) and concentrated hydrochloric acid (3 WO 97/27180 PCT/US97/01610 266 mL) was stirred at ambient temperature for 3h, neutralized with 2N sodium hydroxide, and extracted with diethyl ether. The extracts were dried (MgSO 4 filtered, and concentrated in vacuo to give 0.040 g yield of the crude product, which was used without further purification. MS: ES+, 550 (M+Na) and ES-, 562 (M+C1).
C.
A solution of the amine (0.14 g, 0.27 mmol), methylchloroformate (0.023 mL, 0.30 mmol) and Et 3
N
(0.05 mL, 0.36 mmol) in dichloromethane (2 mL) was stirrred at ambient temperature under a nitrogen atmosphere for 18h. The volatiles were removed in vacuo, and the residue was purified by reverse phase preparative HPLC to give a tan oil. Lyophilization gave 0.012 g of the product as a white solid. MS: ES+,608 H NMR (CDC13) 1.71 1H); 1.96 (m, 1H); 2.11 1H); 2.26 2.71-3.05 8H); 3.50 3H); 3.65 1H); 3.82 1H); 4.00-4.39 5.28 1H); 5.43 1H); 6.40 7.08-7.33 (m, 14H).
WO 97/27180 WO 9727180PCTIUS97/01610 26'7 Exam-ple 91 Synthesis of Compoound 239 phosphazene base. THF, -78 'COH H H N H OH oF Ho
)I
1 ~08 isopropanoklonc. HCI it Prepared as described in Example 74 with the exception that water, rather than 1.0 N HC1 was used to quench the reaction. MS of the acetonide 660 MS of the product 644 1 HNM'R of the product (CDC1 3 d 1.68 (mn, 3H), 2.07 (mn, 3H), 2.54 (in, 2H), 2.92 (mn, 6H), 3.43 (mn, 1H), 3.78 (mn, 1H), 4.00 (mn, 2H), 4.50 (mn, 1H), 5.34 (in, 1H), 6.10 (mn, 1H), 6.70 (mn, 1H) 7.24 (mn, 18H) Exainole 92 Synthesis of Coinoound 238 isopropanol/conc. MCI rt WO 97/27180 WO 9727180PCTIUS97/01610 -268 Prepared as described in Example 74 with the exception that water, rather than 1.0 N HC1 was used to quench the reaction. MS of the acetonide 684 (M+Na).
MS of the product 644 (M+Na) I HNMR of the product (CDC1 3 d 1.62 (mn, 3H), 2.00 (mn, 3H), 2.50 (mn, 2H), 2.80 (mn, 6H 3.30 (mn, 2H), 4.00 (mn, 2H), 4.34 (mn, 1H), 5.33 (mn, 1H), 6.14 (mn, 1H), 6.30 (mn, 1H), -7.24 (mn, 18H) Synthesis of Coinound-240 phosphazene base. THF, -78 9H ~H H O -Q~pNH N NC N 0 0H0 0 1 08 isopropanollconc. HCI it Prepared as described in Example 74 with the exception that water, rather than 1.0 N H1-i was used to quench the reaction. MS of the acetonide 691 (M+Na).
MS of the product 651 (M+Na) 1HNMR of the (CDC1 3 d 1.66 (in, 3H), 2.08 (in, 3H), 2.59 (mn, 2H), 2.95 (mn, 6H), 3.40 (mn, 1H), 3.85 (in, 4.14 (in, 2H), 4.27 (in, 1H), 5.32 (in, 1H), 6.22 (mn, iH), 6.73 (mn, 1H), 7.25 (mn, 18H).
WO 97/27180 WO 9727180PCT[US97/01610 269 Example 94 S nthesis of Compound 241.
Q~~y phosphaz ene base, THF, -78 0 COH H0- N H
HHO
0 H isopropanol/conc. HCI rt Prepared as described in Example 74 with the exception that water, rather than 1.0 N HC1 was used to quench the reaction. MS of the acetonide =645 (M4+1).
MS of the product 627 (M+Na) 1HNMR of the product (CDC13): d 1.70 (in, 3H), 2.00 (mn, 3H), 2.58 (mn, 2H), 2.97 (in, 6Hl), 3.40 (mn, 1Hl), 3.85 (mn, 1Hl), 4.10 (in, 2H), 4.32.(in, 1H), 5.33 (mn, 1H), 6.30 (in, 1Hl), 6.80 (mn, 1Hl), 7.25 (mn, 17H), 8.01 (in, 1H).
WO 97/27180 PCT/US97/01610 270 Example Synthesis of Compound 208 Ph Ph A. NaH, DMF, F3CqS o Ph B. LiOH, THF, DME Ph PhCHN C. TBDMSCI, Im. PhCHNO H
OH
OH
HN.
8 IPh h D. EDC, HOBT, H PsCH F CH30(CO)CI, DIEA O0 N7OH 0 0
A.
The lactam (1.20 g, 2.69 mmol, 1 eq) was dissolved in anhydrous dimethylformamide (8 mL) under Argon and cooled with an isopropanol dry ice bath to -40 oC. A solution of sodium bis(trimethylsilyl)amide (1.OM in THF, 2.69 mL, 2.69 mmol, 1 eq) was added dropwise via syringe and the reaction was stirred for 15 min.
maintaining the bath temp between -40 -50 °C.
3(R)-(phenylmethyl)-3(2H)-furanone Med. Chem., 1994, Vol. 37, No. 21, 3443-51; 1.00 g, 2.96 mmol, 1.1 eq) was added as a solid and the reaction was stirred vigorously for 10 min. and then quenched with several drops of glacial acetic acid. The reaction mixture was evaporated in vacuo to a residue and partitioned between ethyl acetate, saturated aqueous brine, and water. After separating the layers, the aqueous layer WO 97/27180 PCT/US97/01610 271 was back-extracted with ethyl acetate. The combined organic layers were washed with saturated aqueous brine, dried over anhydrous magnesium sulfate, evaporated in vacuo and purified by flash silica gel chromatography eluting with ethyl acetate hexane Fractions containing the alkylated lactam were combined, evaporated in vacuo to provide 0.883 g (52 of product as a foam. MS (ESI): M+Na 657.
B.
The butyrolactone (1.202 g, 1.89 mmol, 1 eq) from step A was dissolved at ambient temperature in dimethoxyethane (20 mL) and cooled with an ice water bath. Aqueous lithium hydroxide (1.0 N, 4.75 mL, 4.75 mmol, 2.5 eq) was added via pipette and the mixture was stirred for 0.5 h. The reaction was warmed to ambient temperature and stirred for an additional 1 h.
Aqueous citric acid (10% w/v) was added to reach an acidic pH and the mixture was evaporated in vacuo. The residue was partitioned between ethyl acetate diethyl ether and aqueous citric acid (10% After separating the layers, the aqueous layer was backextracted with ethyl acetate. The combined organic layers were washed with water, saturated aqueous brine, dried over anhydrous magnesium sulfate, evaporated in vacuo and dried under high vacuum to provide the acid (1.32 g, 106%) as a foam. MS (APCI): M 1 651.
C.
The acid (1.28 g, 1.97 mmol, 1 eq) from step B in 5 mL anhydrous dimethylformamide under Argon was combined WO 97/27180 PCT/US97/01610 272 with imidizole (1.472 g, 21.6 mmol, 11 eq) followed by tertbutyldimethylsilyl chloride (2.96 g, 19.7 mmol, eq) and stirred at ambient temperature for 16 h. The reaction was quench by addition of methanol (15 mL) and stirred for an additional 45 min. Aqueous lithium hydroxide (1.0 N, 2.0 mL, 1 eq) was added and the mixture was evaporated in vacuo. The residue was partitioned between ethyl acetate and aqueous sodium hydrogen sulfate (1.0 After separating the layers, the aqueous layer was back-extracted with ethyl acetate. The combined organic layers were washed with saturated aqueous brine, dried over anhydrous magnesium sulfate, evaporated in vacuo and dried under high vacuum to provide the silyl protected acid (1.46 g, 97%) as a foam. MS (APCI): M 1 766.
D.
The silyl protected acid (1.32 g, 1.72 mmol, 1 eq) from step C in anhydrous dimethylformamide (7 mL) under Argon was treated consecutively with diisopropylethyl amine (0.316 mL, 1.81 mmol, 1.05 eq), 1hydroxybenzotriazole (0.244 g, 1.81 mmol, 1.05 eq), (lS,2R)-(-)-l-amino-2-indanol (0.283 g, 1.90 mmol, 1.1 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.347 g, 1.81 mmol, 1.05 eq). After stirring at ambient temperature for 3 h, the reaction mixture was evaporated in vacuo, and partitioned between ethyl acetate, saturated aqueous brine, and water. After separating the layers, the aqueous layer was back-extracted with ethyl acetate. The combined organic layers were washed with saturated aqueous WO 97/27180 PCT/US97/01610 273 brine, dried over anhydrous magnesium sulfate, evaporated in vacuo and purified by flash silica gel chromatography eluting with ethyl acetate hexane Fractions containing the product were combined, evaporated in vacuo and dried under high vacuum to provide the protected amide (1.06 g, 69%) as a foam. MS (ESI): M+Na 920.
E.
The protected amide (1.035 g, 1.15 mmol, 1 eq) from step D was dissolved in trifluoroacetic acid (15 mL) and stirred under Argon for 15 min. The reaction was evaporated in vacuo and trituratated with diethyl ether/hexane. After decanting the mother liquor, the residual solid was dried under high vacuum to provide a partially deprotected product. The crude material was dissolved again in trifluoroacetic acid (15 mL) and stirred for 20 min. under Argon. The reaction mixture was evaporated in vacuo to a residue and triturated with hexane/diethyl ether. The slurry was filtered, washed with hexane and dried under high vacuum to provide the deprotected amine (0.607 g, 83%) as a trifluoroacetic acid salt. MS (ESI): M+1 542.
F.
The amine (0.025 g, 0.038 mmol, 1 eq) from step E was combined with diisopropylethylamine (0.0146 mL, 0.038 mmol, 2.2 eq) in dichloromethane (1.5 mL) under Argon.
The solution was treated with methylchloroformate (0.0028 mL, 0.0362 mmol, 0.95 eq). After stirring for approximately 10 min., the reaction mixture was applied WO 97/27180 PCT/US97/01610 274 directly to a 20x20 cm (500 uM, silica gel GF) preparative thin layer chromatography plate and eluted with 95:5 dichloromethane methanol. The product band was removed from the plate and the product was washed from the silica gel with 85:15 dichloromethane methanol (10 mL). The solution was evaporated in vacuo, triturated with hexane, evaporated in vacuo, and dried under high vacuum to provide the carbamate as a white solid (0.0164 g, 72 The product was lyophilized from acetonitrile water MS (APCI): M Na 622. H NMR (CDC13 NaOD): 1.66 1H); 1.90 3H); 2.29 1H); 2.64 1H); 2.92 7H); 3.18 1H); 3.40 1H); 3.56 3H); 3.66 1H); 3.85 1H); 3.98 1H); 4.26 1H); 5.27 1H), 6.07 1H, 7.14 6H); 7.28 8H).
Example 96 Synthesis of Compound 236 Ph Ph I Ph Ph
TFA*H
2 H H OH CF 3
SO
2 NH O H
S(CF
3
S
2 2 0 DIEA o Yr The aminomethyl pyrolidinone (0.025 g, 0.038 mmol, 1 eq) was combined with diisopropylethylamine (0.0146 mL, 0.038 mmol, 2.2 eq) in dichloromethane (1.5 mL) and cooled to -78 °C with a dry ice acetone bath. The solution was treated with trifluoromethane sulfonic anhydride (0.0064 mL, 0.038 mmol, 1 eq) in dichloromethane (0.5 mL). The reaction mixture was WO 97/27180 PCT/US97/01610 275 then allowed to warm to room temperature and applied directly to a 20x20 cm (500 uM, silica gel GF) preparative thin layer chromatography plate and eluted with 95:5 dichloromethane methanol. The product band was removed from the plate and the product was washed from the silica gel with 85:15 dichloromethane methanol (10 mL). The solution was evaporated in vacuo to a residue and lyophilized from acetonitrile water to provide the desired product as a white lyophile (0.008 g, 31 MS (APCI): M Na 696. H NMR (CDC13 NaOD): 1.64 1H); 1.98 3H); 2.22 1H); 2.72 1H); 2.91 8H); 3.47 1H); 3.78 1H); 3.97 1H); 4.09 1H); 4.31 1H); 5.23 1H); 6.17 1H, 7.21 14H) Example 97 Synthesis of Compound 211 0 Ph 0 0,O"N Ph PhH Ph
TFA*H,
2 N H OH O N
OH
8 O DIEA a The aminomethyl pyrolidinone (0.030 g, 0.046 mmol, 1 eq) was combined with diisopropylethylamine (0.0175 mL, 0.10 mmol, 2.2 eq) and 3-(R)-hydroxy-tetrahydrofuran-Nhydroxysuccinimide carbonate (W093-US8458, 0.016 g, 0.046 mmol, 1 eq) in dichloromethane (1.5 mL) and allowed to stir for 16 h at ambient temperature. The dichloromethane was removed in vacuo and replaced with acetonitrile (2 mL). The mixture was heated at reflux WO 97/27180 PCT/US97/01610 276 for 20 min. and then cooled and evaporated in vacuo.
The residue was dissolved in dichloromethane mL), applied directly to a 20x20 cm (500 uM, silica gel GF) preparative thin layer chromatography plate and eluted with 9:1 dichloromethane methanol. The product band was removed from the plate and the product was washed from the silica gel with 85:15 dichloromethane methanol (10 mL). The solution was evaporated in vacuo to a residue and lyophilized from acetonitrile water to provide the desired product as a white lyophile (0.022 g, 73 MS (ESI): M Na 678. H NMR (CDC13 NaOD): 1.65 1H); 1.93 5H); 2.32 1H); 2.65 1H); 2.90 7H); 3.22 1H); 3.37 1H); 3.54 2H); 3.79 4H); 3.97 1H); 4.22 1H); 5.11 1H); 5.27 1H); 6.34 1H, 7.22 14H).
Examnle 98 Synthesis of Compound 215 1. NaH /THF MeO O *N OHO N H- OH OH 2. HCI The starting cyclic urea was obtained following procedures outlined in Examples 11 and 12. Coupling with the epoxide followed the protocol detailed in Example 24.
WO 97/27180 PCT/US97/01610 277 Example 99 Synthesis of Compound 242 bCO H- 0 1. TFA N Y) N o o0 2 1 0.3g of the protected intermediate obtained in Example 98 was treated with 10-mL of TFA over 5 h at room temperature. The reaction was quenched by removing the TFA, and the resulting crude treated with excess of sodium carbonate in methanol/water for 10 minutes. The solvents were removed, product extracted between ethyl acetate/water, organics combined, dried with magnesium sulfate, removed in vacuo, and purified by preparative HPLC, resulting in 0.15g of product 2. H NMR (CDC13, 300 MHz) 5 8.10 (1H, d, 7.24 (10H, m), 7.05 (5H, 5.28 1H), 4.10 (1H, t, 3.97 (1H, t, 3.53 (1H, 3.39 (2H, 2.95 2.69 2H), 2.54 (1H, dd), 2.17 (1H, 1.92 (1H, 1.78 1H). Low resolution MS m/e 514.1 (M+H m/e 536.2 (M+Na) WO 97/27180 WO 9727180PCT/US97/01610 278 E x amnLelO0 Svnthesis of Compound 243 OHHF R-halide
H
08 0 08 Ak solution of 20 mg (0.039 Inmol) of the urea obtained Example 99 in I mL DMF was treated with potassium tbutoxide (26.3 mg, 0.234 inmol, 6 equiv) and equilibrated at room temperature for 10 min. Next, 6.3 mg of 3-picolyl chloride in 1 mL DMF was added and the reaction quenched after 20 min. Solvent were then removed and the residue purified on preparative
RP
HPLC resulting in 14.2 mg (60.20%) of the product. 1
H
NMR (d6-acetone, 400 MHz) 5 8.57 1H, 8.42 1H), 8.01 1H, 7.80 1H, 7.20 Cm, 14H1), 6.92 1H1, 5.23 (in, 1H), 4.29 (d, 111, J=16.2), 4.29 (mn, 1H), 4.11 Cd, 1H, J=16.2), 3.98 (in, 2H), 3.48 (dd, 1H), 3.18 (mn, 2H), 3.00 (in, 4H1), 2.75 3H), 1.93 (mn, 1.88 (mn, 1H1), 1.66 (mn, 1H1).
Low resolution MS ni/e 605.4 mle 627.4 (M+Na WO 97/27180 WO 9727180PCT[US97/01610 279 ExAa-le 1 synthesis of Compopund 244 H KOtBu IIR-halide 7 H This compound was synthesized using the protocol outlined for Example 100 starting from 51 mg (0.1 MM) of cyclic urea and 3-methylbenzyl bromide (18.5 mng, 0.1 mmiol, 1. equiv), resulting in 6.2 mng of the product after preparative HPLC purification. 1H NMR (d6-DMSO, 300 MHz) 6,_7.68 (1H, d, 7.24 (19H, in), 5.18 (1H, in), 4.26 (1H, mn), 4.16 (1H1, d, J=15.7), 4.01 (1H, d, J=15.7), 3.79 (2H, in), 3.33 (1H, in), 3.05 (6H, in), 2.78 (mn, 211), 2.62 (2H, mn), 2.24 3H), 1.80 mn), 1.38 (1H, in). Low resolution MS rn/e 618.2 Example 102 Synthesis of Comnound 245 H OH KOtBu FOH H R-halide mH YN -y o-N3 WO 97/27180 WO 9727180PCT/US97/01610 -280 This compound was synthesized using the protocol outlined for Example 100 starting from 51. mg (0.1 inN) of cyclic urea and 3-fluorobenzyl bromide (18.9 mng, 0.1 mmiol, 1 equiv), resulting in 7.6 mg of the product after preparative HPLC purification. IH NMR (d6-DMSO, 300 MHz) 5 7.71 (1H, 7.24 (19H, in), 5.18 (1H, mn), 4.28 (1H, in), 4.19 (1H, d, J=15.7), 4.03 (1H, d, J=15.7), 3.82 (2H, in), 3.33 (1H, dd), 3.05 (6H, mn), 2.80 (mn, 2H), 2.59 (2H, in), 1.79 (1H, in), 1.38 (1H, in).
Low resolution MS W/e 622.1 Exainvle 103 Synthesis of Comp~ound 262 Nr Obtained following the protocol outlined for Example 100 using 2-picolyl chloride.
is LC/MS-MH( 605.
WO 97/27180 WO 9727180PCTIUJS97/01610 281 Example 104 synthesis of Comipound 213 K~tBu R-halide Obtained following the protocol outlined for Example 100 using 3,4,5-trimethoxybenzyl chloride.
1H NM~R (DMSO)d 6 1.35 (t,1H),1.78 2.45 (m,2H), 2.62 3.8 4.1 4.28 5.18 6.45 6.93-7.38 (m,16H) 7.68 LC/MS-MH 694.
Exam-Ole 105 Synthesis of Comoound 246 KOtBu R-hahide obtained following the protocol outlined for Example 100 using 4-amidobenzyl chloride.
1 HNMR (DMSOd 6 1.35 (t,1H),l.78 (t,1H),2.45 (m,2H),2.62 (in, 3.8 4.1(q,2H), 4-28 WO 97/27180 WO 9727180PCTIUS97/01610 282 5.18 6.45 (s,2H), 7.68 LC/MS-MH+ 694.
Example 106 synthesis of Comipound 257 6.93-7.38 (m,16H) MCjQ$Q_ 0 1. bromomethylacrylic acid! 2 eq. NaH, TI-W 03~ b. M2S' 3. EDCI, HOBT,H OCNH--i 9 0 ONH-t-bu Following the procedure outlined in Example 21 the desired ketoamide was obtained as a white fluffy solid after purification on reversed phase HIPLC. M+H: 504 1 H NMR: 1.38 and 1.48 l.8-3.0(ca 7H,m), 3.72 and 3.73 3.5(1H,m), 3.8(lH,m), 4.0(2H,m), 4.2- 4.8(3H,m) 7.2-7.4(5H,m). Note: Complex NMR signals due to rotational isomers, diastereomers and ketone-hydrate equillibria.
Example 107 Synthesis of Compound 258 Ph 0 Ph,) OH 0 HOBT/EDCI/NM M/DMAP
HK
H
WO 97/27180 PCT/US97/01610 283 The procedure was followed as described in Example 21 except that instead of thioproline-t-butylamide being coupled to the ketoacid 1, thioproline-dimethyl propargylamide 2 was used. This compound was made by treating a 0 °C solution of N-BOC-4-thio-L-proline (Sigma, 2.0 g, 8.6 mmol) in THF (40 mL) with diisopropylethylamine (4.5 mL, 26 mmol) followed by dropwise addition of isobutyl chloroformate (1.1 mL, 8.6 mmol) via syringe. The reaction was stirred for minutes at 0 °C before the dropwise addition of 1,1-dimethylpropargylamine (Aldrich,-1.0 mL, 8.6 mmol).
After stirring for 17 h at room temperature, the reaction was concentrated in vacuo. Ethyl acetate mL) and water (35 mL) were added to the residue and the layers were partitioned. The organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. The crude residue was then dissolved in dichloromethane (20 mL) and treated slowly with trifluoroacetic acid (20 mL). The reaction was stirred for 24 h before being diluted with ethyl acetate mL) and carefully neutralized with 10% sodium carbonate to pH 7. The layers were partitioned and the organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. Flash chromatography over silica gel (1:1 hexane:ethyl acetate) gave amide 2 as a white foam. MS 199 Coupling of ketoacid 1 (300 mg, 0.854 mmol) with amide 2 (170 mg, 0.854 mmol) gave ketoamide 3 (84 mg, 0.211 mmol, after preparatory silica gel TLC (3:1 ethyl acetate:hexane). MS 532 554 (M+Na); HNMR (CDC1 3 d 1.66 3H), 1.69 3H), 1.94 (m, 2H), 2.37 1H), 2.51 3H), 2.92 1H), 3.21 (m, WO 97/27180 WO 9727180PCTIUS97/01610 284 2H), 3.52 Cm, 1H), 3.83 (in, 1H), 4.22 (mn, 2H), 4.44 (m, IH) 4. 81 1H) 5. 00 (in, 1H) 6.57 1H) 7. 1Cm 4H), 7.22 (mn, 6H).
Examplie 108 Synthesis of Compound 263 sodium hydride, DMFOH H 9 2 NH2.)
N
ON,
0 8 isopropanollconc. MCI nt Prepared using the procedure outlined in Example 24.
The acetonide was purified by column chromatography: 65/35 hexane/ethyl acetate. MS: M+Na 643. The product was purified by column chromatography: 40/60 hexane/ethyl acetate. MS: M+Na =603 1 H NMR (CDC1 3 1-O05(m, iH); 1.10-1.40(m, 6H); 1.50-1.75(m, 4H); 1.80- 2.00(m, 2H); 2.45(m, 1H); 2.80-3.10Cm, 4H); 3.20(m, 2H); 3.30Cm, 111); 3.45Cs, 1H); 3.65(m, 1H1); 3.80Cm, 1H); 3.90(m, 1H); 4.25(m, 1H); 4.60(m, 1H); 5.27(m, 1H); 6.00(d, iH); 7.10-7.40(m, 14H).
WO 97/27180 PCT/US97/01610 285 Example 109 Synthesis of Compound 206
A.
A solution of 22.3g (0.147 mol, 1 equiv) of Amino-3-phenyl-l-propanol in 30 mL THF, cooled to 0 OC, was treated with 25.5 mL (0.147 mol, 1 equiv) of DIEA, followed by addition of 11.7 mL (0.147 mmol, 1 equiv) of chloroacetyl chloride. After 1 hr at room temperature, 18.0g (0.16 mol) of potassium -tertbutoxide was added at 0 OC, the reaction warmed up to room temperature and allowed to proceed for 15 min.
Solvents were then removed and the crude residue partitioned between ethyl acetate/water, organics dried over MgSO 4 resulting in 23.8g of the desired product. H NMR (CDCL 3 300 MHz) 6 7.20 5H), 6.67 1H), 4.15 2H), 3.75 1H), 3.86 (dd, 1H, J=11.6, 3.55 (dd, 1H, J=11.6, 2.82 2H).
Low resolution MS m/e 192.1 (M+H
B.
A solution of 0.477g (2.5 mmol, 1 equiv) of the morpholinone above in 1 mL of anhydrous DMF was treated with 12 mg (0.5 mmol, 0.2 equiv) of sodium hydride WO 97/27180 PCTIUS97/01610 286 at O'C. The reaction was continued at room temperature for 10 min, and then cooled down to O'C, followed by addition of 0.813g (2.5 mmol, 1 equiv) of epoxide in 1 mL DMF. The reaction was then carried out at 50'C for 5 h. Following ethyl acetate/water extraction, the organics were combined and dried resulting in 1.18 g of crude product, used further without purification. Low resolution MS m/e 539.0 (M+Na
C.
A solution of 1.18g (2.287 mol, 1 equiv) of the above crude in 4 mL anhydrous THF was treated with 0.44g (3.43 mol, 1.5 equiv) of DIEA, followed by 0.907g (3.43 mmol, 1.5 equiv) of TBDMS triflate. After 1 h at room temperature, the product was purified on silica gel (Rf=0.
26 1:3 ethyl acetate/hexane), yielding 0.85g of the TBDMS ether H NMR (CDCL 3 300 MHz) 5 7.50 2H, 7.24 5H), 6.97 2H, 4.40 1H), 4.21 1H, 4.17 1H, 3.82 3H), 3.65 2H), 3.54 1H), 3.35 1H), 3.17 1H), 3.00 4H), 2.77 1H), 2.21 1H), 1.79 1H), 1.57 5H), 1.24 1H), 1.03 1H), 0.86 9H), 0.05 3H), 0.02 3H). Low resolution
MS
m/e 653.1 (M+Na m/e 631.1 (M+H
D.
A solution of 0.12g (0.19 mmol, 1 equiv) of the precursor above in 1.5 mL THF was cooled to -78'C and added 0.25 mL (0.25 mmol, 1.3 equiv) lithium bis(trimethylsilyl)amide (lM solution in THF). After min, 0.029 mL (0.248 mmol, 1.3 equiv) of benzyl bromide WO 97/27180 PCT/US97/01610 287 was added and reaction allowed to proceed at room temperature for additional 1 h. Purification on silica gel (mixture of diastereomers, Rf=0.
46 0.51 in 1:3 ethyl acetate/hexane) provided 44 mg of the TBDMS-protected product. Low resolution MS m/e 1464.6 m/e 721.1
-E.
A solution of 40 mg of the silylated product above in 0.3 mL THF was treated with 0.3 mL of 1M TBAF in THF for 25 min at room temperature and purified on a silica column, resulting in 30 mg of the final product.
Rf=0.
38 and 0.34 (2/5/0.3 ethyl acetate: hexane: methanol). H NMR (CDCL 3 300 MHz) shows both diastereomers and integrates as expected. Low resolution MS m/e 629.3 Example 110 Synthesis of Compound 205 WO 97/27180 PCT/US97/01610 288 A solution of 0.12g (0.19 mmol, 1 equiv) of the compound prepared in Example 109C was dissolved in mL THF and was treated with 0.30 mL (0.30 mmol, equiv) of lithium bis(trimethylsilyl)amide (1M solution in THF) at -78 After 20 min, 0.023 mL (0.266 mmol, 1.4 equiv) of allyl bromide-was added, reaction allowed to warm up to the room temperature and carried out for additional 1 h. The reaction was then quenched with aqueous ammonium chloride and both diastereomers separated on a silica gel. The (lower) Rf=0.50 diastereomer (1:3 ethyl acetate/hexane) was then treated with 10-fold excess of TBAF (1M in THF) for min at room temperature, followed by another silica purification, which provided 14 mg of the desired allylated product. H NMR (CDCL 3 300 MHz) 6 7.73 (d, 2H, 7.24 5H), 6.99, 1H, 5.84 1H), 5.12 2H), 4.27 1H), 4.10 1H), 3.86 3H), 3.84 3H), 3.58 1H), 2.8-3.3 7H), 2.62 2H), 2.09 1H), 1.60 6H), 1.24 2H).
Low resolution MS m/e 579.3 (M+Na m/e 1135.4 (2M+Na+).
WO 97/27180 PCT/US97/01610 289 Example 111 Synthesis of Compound 207 qDMF O OH 0 02 ONC ->RT o 1 2
NO
2 A solution of 0.092g of the morpholinone described in Example 20 (0.35 mmol, lequiv) in 1.5 mL anhydrous DMF was cooled to 0 OC and added 9.6 mg (0.4 mmol, 1 equiv) of NaH. After 1/2h 0.13g (0.32 mmol) of the epoxide 2 was added and reaction carried out at room temperature for 10 h, quenched with IN HClaq, and purified on preparative RP HPLC. Yield 70 mg Low resolution MS m/e 622.1 (M+Na m/e 1221.1 (2M+Na Example 112 Using the methods described by Pennington et al.
and Partaledis et al. (supra), we obtained inhibition constants for the following compounds of this invention: Compound hi (nM) 1 160 2* 180 3* 1,800 >10,000 WO 97/27180 PCT/US97/01610 290 6* >10,000 7 9 8* 9* 10 >10,000 11 >10,000 12 >10,000 13 225 14 16 15 550 16 56 17 115 18 19 3,000 20 21 >20,000 22 600 23 24 350 25 83 26 58 27 3,000 28 1,400 >15,000 31 390 32 160 33 1,100 34 950 130 36 >20,000 37 >20,000 38 17 WO 97/27180 WO 9727180PCTIUS97/01610 -291- 39 600 >20, 000 41 >20, 000 42 330 43 >10, 000 44 120 46 >10, 000 47 50* 100 51* 52* 1,100 54* 12 56* 280 57* 400 58* 5,800 59* 000 170 61* 000 62* 120 63* 200 64* 000 2,900 66* 1,300 67* 3,900 68* >10, 000 69* >10, 000 790 71* 2,500 72* 73* 190 WO 97/27180 WO 9727180PCTIUS97/01610 292 74* 76* 77* 78* 79* 82* 83* 8'7* 88* 91* 92* (isomer 1) 92* (isomer 2) 93 96* 98* 102* 105* 109* ill* 112* 113* 114* 115* 123* (isomer 1) 123* (isomer 2) 124* (isomer 1) 124* (isomer 2) 125* (isomer 1) 1,200 250 560 000 3 0.50 2,500 270 220 12 300 420 4 >10, 000 1,200 >10, 000 250 >10, 000 8, 600 >10, 000 000 >10, 000 300 13 800 1900 400 WO 97/27180 WO 9727180PCT/US97/01610 293 125* 12 6* 127* 128* 129* 130* 131* 131* 132* 133* 133* 208* 209* 210* 211* 212* 213* 214* 215* 216* 217* 218* 219* 219* 219* 220* 221* 223* 224 225* 226* 227* (isomer 2) (isomer 1) (isomer 2) (isomer 1) (isomer 2) (isomer 1) (isomer 2) (isomer 3) 1000 86 92 96 400 100 42 52 24 120 100 2,200 100 5,600 5, 900 3, 100 240 10, 000 1, 000 >10, 000 700 54 330 7 18 370 29 100 WO 97/27180 PCT/US97/01610 294 228* 229* 232* 233* (isomer 1) 233* (isomer 2) 235* 236* 16 28 500 23 1200 270 3.6 Inhibition constant measured at pH Example 113 Using the MT4 cell assay method (supra), we measured the antiviral activity for the following compounds of this invention: Compound ICSO (UM)M 54 83 92 96 123 123 127 130 131 131 132 (isomer 1) 16 9 0.32 0.21 2 0.40 0.90 0.74 0.85 2.4 2.9 0.75 (isomer (isomer (isomer (isomer WO 97/27180 PCT/US97/01610 295 214 2.15 219 (isomer 1) 0.4 219 (isomer 2) 1.7 219 (isomer 3) 220 0.10 223 0.68 224 225 226 227 2.75 228 0.48 229 0.79 232 2.47 233 (isomer 1) 3.7 233 (isomer 2) 1.6 236 The above data show that each of the tested compounds inhibits HIV aspartyl protease.
While we have described a number of embodiments of this invention, it is apparent that our basic constructions may be altered to provide other embodiments which utilize the products and processes of this invention. Therefore, it will be appreciated that the scope of this invention is to be defined by the appended claims, rather than by the specific embodiments which have been presented by way of example.
Claims (53)
1. A compound according to formula I: (I) **t wherein: each Z is R 7 /NX, xY 4 SEC ~T O4: wherein any Z is optionally fused with R6; each X and X' is independently selected from the group consisting of and S(0)2; each Y and Y' is independently selected from the group consisting of -NR2-, >C=C(R2)2 and -N(R2)-CH 2 each R 1 is independently selected from the group consisting of hydrogen; R 6 C 1 -C 6 alkyl; C 2 -C 6 alkenyl; C 2 -C 6 alkynyl; C 3 -C 6 cycloalkyl optionally fused with R C 5 -C 6 cycloalkenyl optionally fused with R6; wherein any member of R 1 is optionally substituted by one or 2 more R -297- 2 each R is independently selected from hydrogen; R C 1 -C 6 alkyl; C 2 -C 6 alkenyl; C 2 -C 6 alkynyl; C 3 -C 6 cycloalkyl optionally fused with R6; C5-C 6 cycloalkenyl optionally fused with R 6 and when two R 2 's are attached to the same geminal atom, the R2's together with their attached geminal atom form a ring system; wherein any member of R 2 is optionally substituted by 3 one or more R each R is independently selected from oxo, OR 9 1 0 N(R 9 2 N(R 9 -X-R N(R 9 -X-OR 9 N(R 9 2, SR, X- 9 9 9 9 R9 O-X-N(R )2 C(O)N(R9)2, halogen, NO 2 CN, COOR and 6 0 4 each R is independently selected from from the 9 9 group consisting of OR; N(R9)2 X-R9; C(O)N(R9)2; R; 15 -C 1 -C 6 alkyl; C 2 -C 4 alkenyl; C 3 -C 6 cycloalkyl optionally fused with R C 5 -C 6 cycloalkenyl optionally fused with R wherein any member of R 4 is optionally substituted by one or more groups independently selected from R 9 or 3 R each R 5 is independently selected from the group consisting of H, OH, O and R 1 6 each R is independently selected from the group consisting of C6-C10 aryl, C3-C8 carbocyclyl and C3-C11 heterocyclyl, wherein said aryl, carbocyclyl or heterocyclyl is optionally substituted with one or more groups selected from the group consisting of oxo, -OR 9 -R (R9 -N(R SR9, -X-R9, -O-X-N(R )2, -R -OR 9 -CN, -C0 2 R 9 -X-N(R 9 (R 9 halogen, -NO 2 and -CF 3 each R is independently selected from the group consisting of hydrogen, OH and 0;- each R is independently selected from the group consisting of hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6-C10 aryl, C3-C8 carbocyclyl, and 4 'a n -298- C3-C11 heterocyclyl; each R is independently selected from the group consisting of hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6-C10 aryl, C3-C8 carbocyclyl, C3-Cl1 heterocyclyl, C1-C10 alkyl substituted C6-C10 aryl C1-C10 alkyl substituted C3-C8 carbocyclyl and C1-C10 alkyl substituted heterocyclyl; wherein any member of R is optionally fused with R 8 and wherein any member 8 of R is optionally substituted by one or more groups 10 independently selected from -OR 8 -N(R 8 -CN, -NO 2 X-R8 -X-N(R8)2, -C(O)OR, -N(R )-XN(R 8 2 or halogen; S: each Q is independently selected from the group consisting of CH and N; each M is independently selected from the group consisting of NH, -NR 2 and each n is independently 1 or 2; each n is independently 1 or 2; each p is independently 1 or 2; each q is independently 1, 2 or 3; and e a c h q is independently 1, 2 or 3; and each G is independently selected from the group consisting of -NR 2 S(0)2, and -C(R 2
2. The compound according to claim 1, wherein: each Y and Y' is independently selected from the group consisting of -NR 2 2 and -N(R 2 )-CH 2 and each R is independently selected from oxo, OR 9 N(R9)2 N(R9)-X-R 9 N(R )-X-OR 9 SR X-R O-X-N(R9)2, C(O)N(R halogen, NO 2 CN, COOR and R 6
3. The compound according to claim 1 having the structure of formula IA: SEC 104 m -299- C. C.. C C. C R\ R12 R R Y XN Z (IA) wherein: 12 each R 12 is independently selected from the group consisting of R6; C1-C6 alkyl optionally substituted with R6; C2-C6 alkenyl; C 2 -C 6 alkynyl; C 3 -C 6 cycloalkyl optionally fused with R6; C5-C6 cycloalkenyl optionally fused with R wherein any member of R 12 is optionally substituted by one or more R 2 15
4. The compound according to claim 1, wherein n is 1. The compound according to claim 1 having the structure of formula II: 7 R 7 R7 R R 7 R
6. The compound according to claim 1 having the structure of formula III: Sn. 300 7 H R The compound according to claim 1, 4 10 wherein: X is -C0(0) or -S and 2 M- Y is 2 )pM.
8. The compound according to claim 1, wherein: :X is or and Y is 2)-P
9. The compound according to claim 1, wherein: X is or and Y is -N(R or -N(R 2)-CH 2 A compound according to formula IV: 7 R 7 H R_ H H Y N N x (IV) R 1 T1O- -301 wherein: X and X' are independently or Y is -(C(R2 -(C(R2 or -N(R2)- CH 2 and each R R R R p and M is independently as defined in claim 1.
11. A compound according to formula V: 7 R R R IR 2Y*- Y N 110 CH2-; R1 0 is O or H2; each R11 is independently H, OH or O, wherein both R11 are not simultaneously hydrogen; Z is a structure of formula VI: I R X' R 4 (VI) (VI) -302- wherein any structure of formula VI is optionally fused with an aryl, carbocyclic or heterocyclic ring and is optionally substituted with 1-3 substituents 2 independently selected from R and each R 1 R 2 R 7 R 4 R 8 p, q, G, M, Q and X' is independently as defined in claim 1.
12. The compound according to claim 11, wherein R 10 and R 11 are 0.
13. The compound according to claim 12, wherein: *q is 1; G is S; and X' is 4
14. The compound according to claim 13, wherein R is t-butylamino. The compound according to claim 12, wherein: X is Y is and S7 R is H.
16. The compound according to claim 11, wherein: X and X' is Y is R 7 is H; R1 0 is H 2 and 11 11 one R1 is H and one R1 is OH.
17. The compound according to claim 16, wherein R 2 within the definition of Y is selected from hydrogen, R or C 1 -C 6 alkyl optionally substituted with R 3 -303-
18. The compound according to claim 17, wherein R 2 within the definition of Y is selected from hydrogen, -N(R or heterocyclyl, which may be optionally benzofused, and wherein said heterocyclyl may be optionally substituted with one or more groups selected 9 9 9 9 from the group consisting of oxo, -OR -R -N(R (R -N(R SR -O-X-N(R 9 2 -R9-OR9, -CN, -CO 2 R 9 -X-N(R 9 (R 9 halogen, -NO 2 and -CF 3
19. The compound according to claim 18, wherein at least one R 2 within the definition of Y is selected from the group consisting of: S, Me 7 I HI 7VY f7 0 H CH 3 N H CH 3 H 3 C Me W\ 7>S jik b A' 9 9 9. 9 9 9 9 9* 9. 9 a 9* 9* 9.9 *4 9 @9 .9 9 6 .dc~ ~4> ~cn -4 ~m ~2j o ~OAIj o~ mz m Z\ P-Z mz z m z P\ -305- OH NH 2 OH HO F F /OH .N N. F F *53*
20. The compound according to claim 17, wherein at least one R 2 within the definition of Y is aryl optionally substituted with one or more groups selected from the group consisting of oxo,.-OR 9 -R -N(R )(R 9 9 9 9 9 9 9 9 -N(R )-X-R 9 SR -X-R 9 -O-X-N(R 9 -R -OR -CN, -C0 2 R 9 -X-N(R 9 halogen, -NO 2 and -CF 3
21. The compound according to claim 17, wherein at 2 least one R2 within the definition of Y is C 1 -Cg alkyl optionally substituted with R 3 Vl, -306-
22. The compound according to claim 21, wherein at least one R within the definition of Y is pyridyl, triazolyl, oxazolyl, isoxazolyl, pyrimidyl, pyrazolyl, pyridazinyl, thiazolyl, imidazolyl, thienyl thiadiazolyl, oxadiazolyl, triazinyl or pyrazinyl wherein said R 3 may be optionally substituted with 1-3 substituents selected from -OR 9 -R 9 -N(R 9 (R 9 9 9 9 9 9 9 9 N(R )-X-R 9 SR -X-R -O-X-N(R 9 -R -OR -CN, -CO 2 R 9 -X-N(R 9 (R 9 halogen, -NO 2 and -CF 3
23. The compound according to claim 21, wherein R 3 within the definition of Y is aryl optionally substituted with 1-3 substituents selected from -OR 9 -R -N(R9)(R -N(R SR -X-R 9 -O-X-N(R )2, -R9-OR 9 -CN, -CO 2 R 9 -X-N(R 9 (R 9 halogen, -NO 2 and -CF 3
24. The compound according to any one of claims 17-23, wherein R is benzyl; and Z is o NX H The compound according to any one of claims 17-23, wherein R 1 is benzyl optionally substituted with 9 9 9 1-3 substituents selected from -OR, -N(R(RCF 3 SR -X-R 9 -R 9-OR 9 -CN, halogen, -NO2, and -CF 3 -307-
26. The compound according to claim 25, wherein Z p. I::
27. The compound according to claim 25, wherein R 1 is benzyl optionally substituted with 1-3 substituents selected from the group consisting of OCH 3 OH and NH 2
28. The compound according to claim 27, wherein Z
29. A compound according to formula V, wherein: i1 Ki 'Ii, /i T *j (V) -308- each R is independently selected from the group consisting of aryl, carbocyclyl and heterocyclyl, wherein said aryl, carbocyclyl or heterocyclyl is optionally substituted with one or more groups selected 9 9 9 9 from the group consisting of oxo, -OR -R -N(R (R 9 9 9 9 9 9 9 -N(R SR -X-R -O-X-N(R -R -OR -CN, -CO 2 R 9 -X-N(R 9 (R 9 halogen, -NO 2 -CF 3 -O-(CH 2 )q-R 6 -O-(CH 2 )q-OR 2,3-methylenedioxy and 3,4- methylenedioxy; and 1 2 3 4 5 7 8 9 10 each X, Y, Z, R R R R R R R R, Q, r" M, n, r, p, q and G is independently as defined in claim 1. .r The compound according to claim 29, wherein R within the definition of Y is selected from hydrogen, R or C 1 -C 6 alkyl optionally substituted with R
31. The compound according to claim 11, wherein: X and X' is Y is -N(R2) R 7 is H; 10 R is H 2 and one R 11 is H and one R 11 is OH.
32. The compound according to claim 11, wherein: X and X' is Y is -(C(R 2 2 M-; M is O; R 7 is H; R1 0 is H 2 and one R 1 1 is H and one R 1 1 is OH. -7 104 NT M -309-
33. The compound according to claim 1, having the structure of formula XII: R Y H R 1 Y- xN- R4 (XII) wherein: X and X' are independently or
34. The compound according to claim 33, wherein R 4 is 1-amino-2-hydroxyindanyl. The compound according to claim 1, having the structure of formula XIII: RH RR H R Y N N Y-.XN X R 9 XIII) wherein: X and X' are independently or
36. The compound according to claim 35, wherein: X' is Y is or and -:353 R 7 is H. -310-
37. The compound according to claim 36, wherein: X is and Y is -(C(R 2 2
38. The compound according to claim 37, wherein R 2 within the definition of Y is selected from hydrogen, R 3 or C1-C 6 alkyl optionally substituted with R 3
39. The compound according to claim 38, wherein R 2 10 within the definition of Y is selected from hydrogen, N(R or heterocyclyl, which may be optionally benzofused, and wherein said heterocyclyl may be .optionally substituted with 1-3 groups selected from the group consisting of oxo, -OR 9 -R 9 -N(R 9 (R 9 99 9 9 9 9 9 15 N(R )-X-R 9 SR -X-R 9 -O-X-N(R 9 2 -R -OR9, -CN, 9 9 9 S-C 2 R -X-N(R halogen, -NO 2 and -CF 3 The compound according to claim 39, wherein at 2 least one R within the definition of Y is selected from the group consisting of: H N IH-C Me H SEC 104 /V eg 9* 0* P\ I Z\ mz P-\ z m Z\ P\ ECZ> -312- -7 NJ H2 S NH 2 I. 4** OH H2 0 NF HO OH OH 7- N~N F 6F F F
41. The compound according to claim 38, wherein at least one R 2within the definition of Y is aryl optionally substituted with one or more groups selected 9 9 9 9 from the group consisting of oxo, -OR -R I -N(R -313- -N(R SR 9 -X-R 9 -O-X-N(R) 2 -R9-OR 9 -CN, -CO 2 R 9 -X-N(R 9 (R 9 halogen, -NO 2 and -CF 3
42. The compound according to claim 38, wherein at 2 least one R within the definition of Y is C 1 -C 6 alkyl optionally substituted with R 3
43. The compound according to claim 42, wherein at least one R 3 within the definition of Y is pyridyl, S* 10 triazolyl, oxazolyl, isoxazolyl, pyrimidyl, pyrazolyl, pyridazinyl, thiazolyl, imidazolyl, thienyl thiadiazolyl, oxadiazolyl, triazinyl or pyrazinyl wherein said R 3 may be optionally substituted with 1-3 9 9 9 9 substituents selected from -OR -R (R N(R9)-X-R 9 SR 9 -X-R -O-X-N(R )2 -R OR 9 -CN, S-CO 2 R 9 -X-N(R 9 halogen, -NO 2 or -CF 3
44. The compound according to claim 42, wherein R 3 within the definition of Y is aryl optionally 9 substituted with 1-3 substituents selected from -OR -R -N(R (R -N(R SR -X-R -O-X-N(R )2, -R9-OR 9 -CN, -CO 2 R 9 -X-N(R 9 (R 9 halogen, -NO 2 or -CF 3
45. The compound according to any one of claims 38-44, wherein: each R 1 is benzyl; and each R not within the definition of Y is 2- hydroxyindanyl.
46. The compound according to any one of claims 38-44, wherein each R 1 is independently selected from benzyl optionally substituted with 1-3 substituents SSEC /r O' -314- selected from -OR -N(R (R SR -X-R 9 -R -OR, CN, halogen, -NO 2 and -CF 3
47. The compound according to claim 46, wherein each R not within the definition of Y is 2- hydroxyindanyl.
48. The compound according to claim 46, wherein each R1 is independently selected from benzyl e* optionally substituted with 1-3 substituents selected from the group consisting of OCH 3 OH and NH 2 9ee9*
49. The compound according to claim 48, wherein 9 15 each R not within the definition of Y is 2- hydroxyindanyl. A compound according to formula XIII, wherein: 7 R R 1 R7 OH R1 YXxN x IN R9 XIII each R 6 is independently selected from the group consisting of aryl, carbocyclyl and heterocyclyl, wherein said aryl, carbocyclyl or heterocyclyl is optionally substituted with one or more groups selected from the group consisting of oxo, -OR 9 -R 9 -N(R 9 (R 9 -N(R 9 )-X-R 9 SR 9 -X-R 9 -O-X-N(R 9 2 -R -OR 9 -CN, -CO 2 R -X-N(R 9 (R 9 halogen, -NO 2 -CF 3 -O-(CH 2 )q-R 6 9 -O-(CH 2 )q-OR 2,3-methylenedioxy and 3,4- methylenedioxy; and "t -315- each X, Y, Z, R, R 2 R R R, R, R R R 9 Q, M, n, r, p, q and G is independently as defined in claim 1.
51. The compound according to claim 50, wherein R 2 within the definition of Y is selected from hydrogen, R or Cl-C 6 alkyl optionally substituted with R 3
52. The compound according to claim 36, wherein: 10 X is and 2 c. Y is .9
53. The compound according to claim 36, wherein: X is -SO 2 and 15 Y is -(C(R 2 2 2 S
54. The compound according to claim 36, wherein X is -S02-; and Y is -N(R 2 The compound according to claim 11, wherein: R is H 2 one R is H and one R 1 1 is OH; and Z is selected from the group consisting of: 2 2 Y and 0 NHtBu 0 NHtBu and R 2 is as defined in claim and R is as defined in claim 1. -316-
56. The compound according to claim 11, wherein Z is selected from the group consisting of H H S H H ,N and N. OS.. 0 0S 0e S. S 0 0 00 R is H 2 and one R11 is H and one R 11 is OH.
57. The compound according to any one of claims 16-32, wherein Z is selected from the group consisting of: N R2 and N 0 NHtBu and R 2 s as defined in claim and R is as defined in claim 1.
58. The compound according to any one of claims 16-32, wherein Z is selected from the group consisting of: -317- and [tBu r r
59. A compound according to formula I, wherein: (I) Z is selected from the group consisting of -X'R 4 -N(R1)-X'-R 4 -N(R1)-N(R1)-X'-R 4 and formula VI; (VI) wherein any structure of formula VI is optionally fused with an aryl, carbocyclic or heterocyclic ring and is optionally substituted with 1-3 members independently selected from R2; and sr SEC -o 104 Ix '/VAT O' -318- each X, Y, Y' R 1 R 2 R 3 R 4 R R 6 R R R 9 Q, M, n, r, p, q and G is independently as defined in claim 1.
60. A compound selected from the group consisting of compound numbers: 1, 2, 3, 4, 7, 8, 9, 13, 14, 16, 17, 18, 20, 23, 24, 25, 26, 32, 35, 38, 44, 46, 47, 48, 49, 50, 51, 52, 53, 54, 62, 63, 72, 76, 78, 80, 82, 83, 91, 92, 94, 95, 96, 101, 102, 109, 10 121, 122, 123, 124, 126, 127, 128, 129, 131, 132, 133, 134, 135, 137, 138, 140, 141, 145, 146, 147, 149, 150, 155, 156, 160, 161, 162, 164, 165, 170, 171, 175, 176, 177, 179, 180, 185, 186, 190, 191, 192, 194, 195, 200, 201, 208, 219, 220, 228 and 264, as shown below in 15 Tables A, B, C, and D: TABLE A OH .A Z Cmpd. No. A Z 1 Ph 3 0 1N rIN.OMe 00 i r C S^ ~i~O u -319- e.* 320 a a a. P h 321 Ph N IiO~ H 3 C N 0 00 38 Ph~ HN O~N H 2 N N 0Me 00 46 Ph O-~Me NN[I 0*0 0 0 47 Ph IH N.ti u S. -Ph P h N 0 H N H 0 NHtBu T O 322 323 S S S 5.5 S S 324 -325- 9 S S *5 S 135 H2 'IN 0 NHtBu 137 0 NHtBu 138 H,, "'H H 2 N 0 NHtBu 140 H,, HfH N 0 NHtBu 141 H. 8, H CN N 0 NHtBu 145 N elpNH2~ 326 C C S *SC*P* S. .4 CS C .5 S C C. 4*CC C. C OC Ir U ~i V 327 00 to SEC 1 RAI 10 4 u T* 328 fl.. S 329 A S.EC 0 /VJT O 330 195 ~~NH2 200 ~NH2 N. 201 ~~NH2 N S. S S S *5 TABLE B OHR A -331 S. S S S. .55* S* S. 5* S S. 332 C 333 TABLE C t p Cmpd No. A z 0 0 NHtBu 102 Ph P h 0 NHtBu 0 109 -Ph S Ph 0 NHtBu 0 p. p p ap TABLE D OH IZ -334- 122 Ph Bn H 0 N_ 0 0
61. The compound of claim 60, selected from the group consisting of compound numbers: 2, 7, 8, 9, 14, 18, 20, 25, 26, 32, 38, 45, 47, 48, 49, 50, 51, 53, 54, 62, 63, 72, 82, 83, 91, 92, 94, 95, 96, 123, 126, 140, 141, 219, 220, 228 and 264.
62. The compound of claim 61, selected from the group consisting of compound numbers: 7, 8, 9, 20, 50, 51, 53, 54, 82, 83, 92, 94, 96, 219, 220, 228 and
264. 63. A pharmaceutical composition comprising an amount of a compound according to claim 1 effective in inhibiting aspartyl protease and a pharmaceutically acceptable carrier, adjuvant or vehicle. 64. The pharmaceutical composition according to claim 63, wherein said pharmaceutical composition is orally administrable. The pharmaceutical composition according to claim 63, further comprising one or more additional agents selected from the group consisting of other anti-viral agents and immunostimulators. 66. The pharmaceutical composition according to claim 65, wherein said other anti-viral agent is a -335- protease inhibitor or a reverse transcriptase inhibitor. 67. The pharmaceutical composition according to claim 66, wherein said protease inhibitor is a HIV protease inhibitor. 68. The pharmaceutical composition according to claim 67, wherein said HIV protease inhibitor or inhibitors are selected from the group consisting of VX-478, saquinavir, indinavir, ritonavir, nelfinavir, palinavir, U-103017, XM 412, XM 450, BMS 186318, CPG 53,437, CPG 61,755, CPG 70,726, ABT 378 GS 3333, GS 3403, GS 4023, GS 4035, GS 4145, GS 4234 and GS 4263. 69. The pharmaceutical composition according to claim 66, wherein said reverse transcriptase inhibitor is a nucleoside analog. 70. The pharmaceutical composition according to claim 69, wherein said nucleoside analog is selected from the group consisting of zidovudine (AZT), dideoxycytidine (ddC), didanosine (ddl), stavudine (d4T), 3TC, 935U83, 1592U89 and 524W91. 71. The pharmaceutical composition according to claim 66, wherein said reverse transcriptase inhibitor is a non-nucleoside analog. 72. The pharmaceutical composition according to claim 71, wherein said non-nucleoside reverse transcriptase inhibitor is delavirdine (U90) or nevirapine. -336- 73. The pharmaceutical composition according to claim 63, wherein said pharmaceutical composition further comprises an agent capable of inhibiting the metabolic effects of one or more cytochrome P 450 enzyme subtypes. 74. A method for inhibiting aspartyl protease activity comprising the step of contacting an aspartyl protease with the compound according to claim 1i. 75. A method for reversibly binding an aspartyl protease comprising the step of contacting the aspartyl protease with the compound according to claim 1, said compound being covalently bound to a solid matrix. 76. A method for preventing HIV infection in a mammal comprising the step of administering to said mammal a pharmaceutical composition according to either claim 63 or 64. 77. A method for preventing HIV infection in a amammal comprising the step of administering to said mammal a pharmaceutical composition according to claim 78. A method for treating HIV infection in a mammal comprising the step of administering to said mammal a pharmaceutically effective amount of a pharmaceutical composition according to either claim 63 or 64. 79. A method for treating HIV infection in a mammal comprising the step of administering to said -337- mammal a pharmaceutical composition according to claim The method according to either claim 76 or 78, further comprising the step of administering, to the mammal one or more additional agents selected from the group consisting of other anti-viral agents and immunostimulators via a single or. multiple dose. 81. The method according to claim 80, wherein said other anti-viral agent is a protease inhibitor or reverse transcriptase inhibitor. 82. The method according to claim 81, wherein 15 said protease inhibitor is an HIV protease inhibitor. 83. The method according to claim 82, wherein said HIV protease inhibitor is selected from the group consisting of VX-478, saquinavir, indinavir ritonavir, nelfinavir, palinavir, U-103017, XM 412 XM 450, BMS 186318, CPG 53,437, CPG 61,755 CPG 70,726, ABT 378, GS 3333, GS 3403, GS 4023, GS 4035, GS 4145 GS 4234, and GS 4263. 84. The method according to claim 81, wherein said reverse transcriptase inhibitor is a nucleoside analog. The method according to claim 84, wherein said nucleoside analog is selected from the group consisting of zidovudine (AZT), dideoxycytidine (ddC), didanosine (ddl), stavudine (d4T), 3TC, 935U83, 1592U89 and 524W91. 0SEC 4 O /VYT O< -338- 86. The method according to claim 81, wherein said reverse transcriptase inhibitor is a non- nucleoside analog. 87. The method according to claim 86, wherein said non-nucleoside reverse transcriptase inhibitor is delavirdine (U90) or nevirapine. 88. A method for treating or preventing of viral infection comprising the step of administering to said mammal a pharmaceutical composition according to either claim 63 or 64. 89. A method for treating or preventing HIV 15 related disease effects, including tumors, CMV retinitis, candida infections, maternal fetal transmission, or AIDS related dementia, comprising the step of administering to said mammal a pharmaceutical composition according to either claim 63 or 64. 90. The composition according to claim wherein the additional anti-viral agents are 3TC and zidovudine (AZT). 91. The composition according to claim wherein the additional anti-viral agent is 1592U89. 92. A process for preparing a compound of formula XIV: -339- R1 R6 NH 0 XIV wherein R and R are defined as in claim 1, S10 comprising the steps of: reacting a compound of formula XV: R1 N Bo c O XV S* 1 wherein R is defined as in claim 1, in an inert solvent with a base; reacting the product of step with an aldehyde of R6CHO followed by an optional treatment with a dehyrating agent, wherein R 6 is defined as in claim 1 to give a compound of formula XVI: R1 R6 NBoc 4--O XVI wherein R 1 and R 6 are defined as in claim 1; SSEC ANT 0 -340- reacting the product of step in an inert solvent with hydrogen gas in the presence of an hydrogenation catalyst followed by treatment with an anhydrous acid to give a product of formula XIV. 93. A process for preparing a compound of formula XVII: 10 R 1 NH XVII reacting a compound of formula XVIII: 20 Ri R2 NH R2 o XVIII wherein R 1 and R 2 are as defined in claim 1, in an inert solvent with a base, then bromomethylacrylic acid; reacting the product of step with an oxidizing agent; reacting the product of step in an inert solvent with thioproline t-butylamide and suitable amide-bond coupling reagents to give a product of S formula XVII. -341 94. A process for preparing a compound of formula XIX: C a. C C C C. cc XIX wherein R and r are defined as in claim 1, comprising the steps of: reacting a compound of formula XX c. C p. S C. S C C. C C. XX in an inert solvent with a base, then a bis-leaving group alkane of formula XXI: rL LG XXI wherein LG is selected from halo, arylsulfonate esters and alkylsulfonate esters, and r is defined as in claim 1, to give a product of formula XXII: -342- O 0 XXII wherein R 1 and PG are defined as in formula XX and LG and r are defined as in formula XXI; reacting the product of step in an inert 10 solvent with a base, to give a product of formula XXIII: 0 o *O 15 (NPG rL r wherein R 1 is defined as in claim 1 and PG is a N- 20 protecting group; reacting the product of step in an inert solvent with a reagent suitable for removal of the N- protecting group PG to give a compound of formula XIX. 95. A compound according to formula I: R 1 R\ R R 7 R 5 R Y--x-N nZ R R (I) -343- wherein: each Z is R1 (Q)r R4 wherein Z is optionally fused with R 6 Q, X, Y, R1, R4, R5, R7 and r as as defined in 10 claim 1; and provided that when Y is -NR 2 or p is 1, X is Q is N and r is 1, then X' is not SO 2 96. The compound according to claim 15 having the structure of formula IX: R R RH-- R 1 Yx N N R 4 O0O (IX) wherein: X is DATED this 27th day of July 1998 VERTEX PHARMACEUTICALS INCOPORATED By their Patent Attorneys CULLEN CO. SRAI SEC 104 NT O9
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US08/592777 | 1996-01-26 | ||
| US08/592,777 US5883252A (en) | 1996-01-26 | 1996-01-26 | Aspartyl protease inhibitors |
| US08/724,563 US5945413A (en) | 1996-01-26 | 1996-09-30 | Aspartyl protease inhibitors |
| US08/724563 | 1996-09-30 | ||
| PCT/US1997/001610 WO1997027180A1 (en) | 1996-01-26 | 1997-01-22 | Aspartyl protease inhibitors |
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| AU1758097A AU1758097A (en) | 1997-08-20 |
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| AU17580/97A Ceased AU709239B2 (en) | 1996-01-26 | 1997-01-22 | Aspartyl protease inhibitors |
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| JP (1) | JP2000501111A (en) |
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| PL (1) | PL328223A1 (en) |
| WO (1) | WO1997027180A1 (en) |
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| WO1998052571A1 (en) * | 1997-05-17 | 1998-11-26 | Glaxo Group Limited | Antiviral combinations containing the carbocyclic nucleoside 1592u89 |
| US6420401B1 (en) | 1997-08-22 | 2002-07-16 | Wichita State University | 1,2,5, thiadiazolidin-3-one 1,1-dioxide derivatives |
| AU2012199A (en) * | 1997-12-24 | 1999-07-19 | Vertex Pharmaceuticals Incorporated | Prodrugs of aspartyl protease inhibitors |
| US6337340B1 (en) * | 1998-08-14 | 2002-01-08 | Gpi Nil Holdings, Inc. | Carboxylic acids and isosteres of heterocyclic ring compounds having multiple heteroatoms for vision and memory disorders |
| US6303609B1 (en) * | 1998-11-18 | 2001-10-16 | Dupont Pharmaceuticals Company | Isoxazoline fibrinogen receptor antagonists |
| DE10047110A1 (en) * | 2000-09-22 | 2002-04-18 | Bayer Ag | Optically active 2,5-bisaryl-DELTA · 1 · -pyrrolines |
| AU2004243511A1 (en) | 2003-05-30 | 2004-12-09 | Neuromed Technologies, Inc | 3-aminomethyl-pyrrolidines as N-type calcium channel blockers |
| BRPI0417697A (en) * | 2003-12-15 | 2007-04-03 | Schering Corp | heterocyclic aspartyl protease inhibitors |
| JP2005232103A (en) * | 2004-02-20 | 2005-09-02 | Nagase & Co Ltd | Optically active vicinaldiamine and method for producing the same |
| GB0412019D0 (en) * | 2004-05-28 | 2004-06-30 | Novartis Ag | Organic compounds |
| US7388008B2 (en) | 2004-08-02 | 2008-06-17 | Ambrilia Biopharma Inc. | Lysine based compounds |
| WO2006114001A1 (en) * | 2005-04-27 | 2006-11-02 | Ambrilia Biopharma Inc. | Method for improving pharmacokinetics of protease inhibitors and protease inhibitor precursors |
| MX2007016185A (en) * | 2005-06-14 | 2008-03-07 | Schering Corp | Macrocyclic heterocyclic aspartyl protease inhibitors. |
| CA2632095A1 (en) | 2005-11-30 | 2007-06-07 | Ambrilia Biopharma Inc. | Lysine-based prodrugs of aspartyl protease inhibitors and processes for their preparation |
| MX2008016399A (en) | 2006-06-20 | 2009-02-03 | Wyeth Corp | Imidazolidinone kv1.5 potassium channel inhibitors. |
| JP5401652B2 (en) | 2006-09-21 | 2014-01-29 | タイメッド バイオロジクス インコーポレイテッド | Protease inhibitor |
| EP2726108B1 (en) | 2011-06-29 | 2018-01-10 | The Trustees of Columbia University in the City of New York | Inhibitor of neuronal connectivity linked to schizophrenia susceptibility and cognitive dysfunction |
| CN103626713B (en) * | 2012-08-29 | 2017-07-04 | 中国科学院上海药物研究所 | Ketone (thioketones) the class compound of nitrogen (oxygen) heterocycle pentane 2, its pharmaceutical composition, preparation method and purposes |
| CN105111198B (en) * | 2015-08-05 | 2018-08-07 | 贵州大学 | Nitro isoxazole splices 2-Pyrrolidone class compound and preparation method and application |
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| US5610294A (en) * | 1991-10-11 | 1997-03-11 | The Du Pont Merck Pharmaceutical Company | Substituted cyclic carbonyls and derivatives thereof useful as retroviral protease inhibitors |
| EP0749421B1 (en) * | 1994-03-07 | 1999-09-15 | Vertex Pharmaceuticals Incorporated | Sulphonamide derivatives as aspartyl protease inhibitors |
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1997
- 1997-01-22 CZ CZ982344A patent/CZ234498A3/en unknown
- 1997-01-22 AU AU17580/97A patent/AU709239B2/en not_active Ceased
- 1997-01-22 JP JP9527124A patent/JP2000501111A/en active Pending
- 1997-01-22 EP EP97904911A patent/EP0882022A1/en not_active Withdrawn
- 1997-01-22 HU HU9901024A patent/HUP9901024A2/en unknown
- 1997-01-22 WO PCT/US1997/001610 patent/WO1997027180A1/en not_active Application Discontinuation
- 1997-01-22 PL PL97328223A patent/PL328223A1/en unknown
- 1997-01-22 CA CA002243121A patent/CA2243121A1/en not_active Abandoned
- 1997-01-22 IL IL12547997A patent/IL125479A0/en unknown
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- 1997-01-24 AP APAP/P/1997/000961A patent/AP844A/en active
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| AP844A (en) | 2000-06-09 |
| HUP9901024A2 (en) | 1999-09-28 |
| PL328223A1 (en) | 1999-01-18 |
| NO983435L (en) | 1998-09-21 |
| WO1997027180A1 (en) | 1997-07-31 |
| AU1758097A (en) | 1997-08-20 |
| AP9700961A0 (en) | 1997-04-30 |
| NO983435D0 (en) | 1998-07-24 |
| IL125479A0 (en) | 1999-03-12 |
| BR9707086A (en) | 1999-04-13 |
| CZ234498A3 (en) | 1998-10-14 |
| EP0882022A1 (en) | 1998-12-09 |
| JP2000501111A (en) | 2000-02-02 |
| CA2243121A1 (en) | 1997-07-31 |
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