AU3554599A - Pyrrolidines as inhibitors of neuraminidases - Google Patents
Pyrrolidines as inhibitors of neuraminidases Download PDFInfo
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- AU3554599A AU3554599A AU35545/99A AU3554599A AU3554599A AU 3554599 A AU3554599 A AU 3554599A AU 35545/99 A AU35545/99 A AU 35545/99A AU 3554599 A AU3554599 A AU 3554599A AU 3554599 A AU3554599 A AU 3554599A
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- carboxylic acid
- pyrrolidine
- acetamido
- hydrogen
- ethyl
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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
- C07D207/30—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 two double bonds between ring members or between ring members and non-ring members
- C07D207/32—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 two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
- C07D207/325—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 two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms with substituted hydrocarbon radicals directly attached to the ring nitrogen atom
- C07D207/327—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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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
- A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D207/16—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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- C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
- C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
- C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
- C07D231/12—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
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- C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
- C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
- C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
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- C07D249/00—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms
- C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
- C07D249/08—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
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- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- 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
- C07D403/04—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 directly linked by a ring-member-to-ring-member bond
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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
- C07D405/04—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 directly linked by a ring-member-to-ring-member bond
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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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- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur 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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- 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
- C07D413/04—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 directly linked by a ring-member-to-ring-member bond
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- 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
- C07D417/04—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 directly linked by a ring-member-to-ring-member bond
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- 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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- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
- C07F9/02—Phosphorus compounds
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
- C07F9/553—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having one nitrogen atom as the only ring hetero atom
- C07F9/572—Five-membered rings
Description
WO 99/54299 PCT/US99/07945 PYRROLIDINES AS INHIBITORS OF NEURAMINIDASES Technical Field The present invention relates to novel compounds, compositions and methods for inhibiting neuraminidase, especially influenza neuraminidase. The invention also contemplates a composition and methods for preventing and treating an influenza infection and processes for making such compounds and synthetic intermediates employed in these processes. Background of the Invention Many disease-causing microorganisms possess a neuraminidase (also known as sialidase) which is involved in the replication process of the microorganism. In particular, viruses of the orthomyxovirus and paramyxovirus groups possess a neuraminidase. Diseases associated with paramyxoviruses include RSV (respiratory syncytial virus-related diseases), pneumonia and bronchiolitis (associated with paramyxovirus type 3) and laryngotracheobronchitis (associated with paramyxovirus type 1). Some of the more important disease causing microorganisms in man and/or animals which possess a neuraminidase include Vibrio cholerae, Clostridium perfringens, Streptococcus pneumoniae, Arthrobacter sialophilus, influenza virus, parainfluenza virus, mumps virus, "1- WO 99/54299 PCT/US99/07945 Newcastle disease virus, fowl plague virus, equine influenza virus and Sendai virus. Mortality due to influenza is a serious problem throughout the world. The disease is devastating to man, lower mammals and some birds. Although vaccines containing attenuated influenza virus are available, those vaccines only provide immunological protection toward a few influenza strains and are less effective in otherwise immunologically compromised populations such as the elderly, young children, and in those who suffer from chronic respiratory illness. The productivity loss from absence due to sickness from influenza virus infection has been estimated to be more than $1 billion per year. There are two major strains of influenza virus (designated A and B). Currently, there are only a few pharmaceutical products approved for treating influenza. These include amantadine and rimantadine, which are active only against the A strain of influenza viruses, and ribavirin, which suffers from dose-limiting toxicity. Mutant virus which is resistant to amantadine and rimantadine emerges quickly during treatment with these agents. Neuraminidase is one of two major viral proteins which protrude from the envelope of influenza virus. During the release of progeny virus from infected cells, neuraminidase cleaves terminal sialic acid residues from glycoproteins, glycolipids and oligosaccharides on the cell surface. Inhibition of neuraminidase enzymatic activity leads to aggregation of progeny virus at the surface. Such virus is incapable of infecting new cells, and viral replication is therefore retarded or blocked. X-ray crystallographic studies and sequence alignments have shown that the residues which directly contact the sialic acid portion of the substrate are strictly conserved in the neuraminidase from all A and B influenza strains. Thus, a compound which binds to the sialic acid binding region of the neuraminidase active site will block the replication of both the A and B strains of influenza virus. Compounds which are influenza neuraminidase inhibitors will be useful for the -2- WO 99/54299 PCT/US99/07945 prevention of influenza infection and will be useful for the treatment of influenza infection. The following references disclose neuraminic acid derivatives with the disclosed utility listed after each reference: L. Von Itzstein, et al., European Patent Application No. EP539204, published April 28, 1993 (antiviral agent); T. Honda, et al., European Patent Application No. EP823428, published February 11, 1998 (sialidase inhibitor; influenza treatment); T. Honda, et al., International Patent Application No. WO98/06712, published February 19, 1998 (sialidase inhibitor; influenza remedy); L. Von Itzstein, et al., International Patent Application No. WO95/20583, published August 3, 1995 (viral neuraminidase inhibitor; influenza treatment); P. Smith, International Patent Application No. WO95/18800, published July 13, 1995 (viral neuraminidase inhibitor); P. Colman, et al., International Patent Application No. WO92/06691, published April 30, 1992 (viral neuraminidase inhibitor); L. Von Itzstein, et al., U.S. Patent No. 5,648,379, issued July 15, 1997 (influenza treatment); P. Reece, et al., International Patent Application No. WO97/32214, published September 4, 1997 (bind to influenza virus neuraminidase active site); and P. Reece, et al., International Patent Application No. WO98/21243, published May 23, 1998 (anti-influenza agent). The following references disclose sialic acid derivatives with the disclosed utility listed after each reference: -3- WO 99/54299 PCT/US99/07945 Y. Ohira, et al., International Patent Application No. WO98/11083, published March 19, 1998 (antiviral agent); Y. Ohira, European Patent Application No. EP882721, published December 9, 1998 (antiviral agent); and B. Glanzer, et al., Helvetica Chimica Acta 74 343-369 (1991) (Vibrio cholerae neuraminidase inhibitor). The following references disclose benzene derivatives, cyclohexane derivatives or cyclohexene derivatives with the disclosed utility listed after each reference: Y. Babu, et al., U.S. Patent No. 5,602,277, issued February 11, 1997 (neuraminidase inhibitors); M. Luo, et al., U.S. Patent No. 5,453,533, issued September 26, 1995 (influenza neuraminidase inhibitor; influenza treatment); Y. Babu, et al., International Patent Application No. WO96/30329, published October 3, 1996 (neuraminidase inhibitor; viral infection treatment); N. Bischofberger, et al., U.S. Patent No. 5,763,483, issued June 9, 1998 (neuraminidase inhibitor); and K. Kent, et al., International Patent Application No. 98/07685, published February 26, 1998 (intermediates for the preparation of neuraminidase inhibitors). C. Kim, et al., International Patent Application No. WO98/17647, published April 30, 1998 discloses piperidine derivatives which are useful as neuraminidase inhibitors. N. Bischofberger, et al., International Patent Application No. WO96/26933, published September 6, 1996 discloses various substituted 6-membered ring compounds which are useful as neuraminidase inhibitors. -4- WO 99/54299 PCT/US99/07945 The following references disclose dihydropyran derivatives which are useful as viral neuraminidase inhibitors: D. Andrews, et al., International Patent Application No. WO97/06157, published February 20, 1997; and P. Cherry, et al., International Patent Application No. WO96/36628, published November 21, 1996. C. Kim, et al., U.S. Patent No. 5,512,596, issued April 30, 1996 discloses 6-membered aromatic ring derivatives which are useful as neuraminidase inhibitors. G. Diana, et al., International Patent Application No. WO98/03487, published January 29, 1998 discloses substituted pyridazines which are useful for treatment of influenza. B. Horenstein, et al., International Patent Application No. WO99/06369, published February 11, 1999 discloses piperazine derivatives which are useful as neuraminidase inhibitors. Y. Babu, et al., International Patent Application No. WO97/47194, published December 18, 1997 discloses substituted cyclopentanes which are useful as neuraminidase inhibitors and treatments for influenza. L. Czollner, et al., Helvetica Chimica Acta 73 1338-1358 (1990) discloses pyrrolidine analogs of neuraminic acid which are useful as Vibrio cholerae sialidase inhibitors. The following references disclose siastatin B analogs which are useful as neuraminidase inhibitors: Y. Nishimura, et al., Natural Product Letters 1 39-44 (1992); and Y. Nishimura, et al., Natural Product Letters 1 33-38 (1992). -5- WO 99/54299 PCT/US99/07945 C. Penn, UK Patent Application No. GB2292081, published February 14, 1996 discloses the use of a neuraminidase inhibitor in combination with an influenza vaccine. An object of the invention is to provide compounds which inhibit neuraminidase of disease-causing microorganisms; especially, viral neuraminidase; and, most especially, influenza neuraminidase. An object of the invention is also to provide compounds which inhibit neuraminidase from both A and B strains of influenza. Another object of the invention is to provide prohylaxis of influenza infection in humans and other mammals. Another object of the invention is to provide treatment of influenza infection in humans and other mammals. Another object of the invention is to provide compounds which exhibit activity against influenza A virus and and influenza B virus by virtue of inhibiting influenza neuraminidase when such compounds are administered orally. Another object of the invention is to provide a compound which can be effectively transported from the plasma into the lung bronchoaveolar fluid of humans and other mammals in order to block the replication of influenza virus in that tissue. -6- WO 99/54299 PCT/US99/07945 Disclosure of the Invention The present invention discloses compounds having Formula I: Y R9 R8 R10 R 6 Rio R"
R
2 -X 1 N R R7 \5 R4R R" R 3 I or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein R 1 is selected from the group consisting of (a) -CO 2 H, (b) -CH 2
CO
2 H, (c) -SO 3 H, (d) -CH 2
SO
3 H, (e) -SO 2 H, (f) -CH 2
SO
2 H, (g) -PO 3
H
2 , (h) -CH 2
PO
3
H
2 , (i) -PO 2 H, (j) -CH 2
PO
2 H, (k) tetrazolyl, (I) -CH 2 -tetrazolyl, (m) -C(=O)-NH-S(O) 2
-R
11 , (n) -CH 2
C(=O)-NH-S(O)
2
-R
11 , (o) -SO 2
N(T-R
1
")R
12 and (p) -CH 2
SO
2 N(T-Rl 1
)R
12 wherein T is selected from the group consisting of (i) a bond, (ii) -C(=O)-, (iii) -C(=O)O-, (iv) -C(=O)S-, (v) -C(=O)NR 36 -, (vi) -C(=S)O-, (vii) -C(=S)S-, and (viii) -C(=S)NR 36 -,
R
11 is selected from the group consisting of (i) C 1
-C
12 alkyl, (ii) C 2
-C
12 alkenyl, (iii) cycloalkyl, (iv) (cyclo alkyl)alkyl, (v) (cycloalkyl)alkenyl, (vi) cycloalkenyl, (vii) (cycloalkenyl)alkyl, (viii) (cycloalkenyl)alkenyl, (ix) aryl, (x) (aryl)alkyl, (xi) (aryl)alkenyl, (xii) heterocyclic, (xiii) (heterocyclic)alkyl and -7- WO 99/54299 PCT/US99/07945 (xiii) (xiv) (heterocyclic)alkenyl; and
R
12 and R 36 are independently selected from the group consisting of (i) hydrogen, (ii) C1-C12 alkyl, (iii) C2-C12 alkenyl, (iv) cycloalkyl, (v) (cycloalkyl)alkyl, (vi) (cycloalkyl)alkenyl, (vii) cycloalkenyl, (viii) (cycloalkenyl)alkyl, (ix) (cycloalkenyl)alkenyl, (x) aryl, (xi) (aryl)alkyl, (xii) (aryl)alkenyl, (xiii) heterocyclic, (xiv) (heterocyclic)alkyl and (xv) (heterocyclic)alkenyl; X is selected from the group consisting of (a) -C(=O)-N(R*)-, (b) -N(R*)-C(=O)-, (c) -C(=S)-N(R*)-, (d) -N(R*)-C(=S)-, (e) -N(R*)-SO 2 -, and (f) -SO 2 -N(R*)- wherein R* is hydrogen, C1-C3 Ioweralkyl or cyclopropyl;
R
2 is selected from the group consisting of (a) hydrogen, (b) C1-C6 alkyl, (c) C2-C6 alkenyl, (d) C3-C6 cycloalkyl, (e) Cs-C6 cycloalkenyl, (f) halo C1-C6 alkyl and (g) halo C2-C6 alkenyl; or R 2 -X- is Y2 wherein Y 1 is -OH 2 -, -0-, -S- or -NH- and Y 2 is -C(=O)- or -C(Raa)(Rbb) - wherein Raa and Rbb are indepedently selected from the group consisting of hydrogen, Cl-C3 loweralkyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, aminomethyl, 1-aminoethyl, 2-aminoethyl, thiolmethyl, 1-thiolethyl, 2-thiolethyl, methoxymethyl, N-methylaminomethyl and methylthiomethyl;
R
3 and R 4 are independently selected from the group consisting of (a) hydrogen, (b) cycloalkyl, (c) cycloalkenyl, (d) heterocyclic, (e) aryl and (f)
-Z-R
14 -8- WO 99/54299 PCT/US99/07945 wherein Z is (v) -C(=S)-, (vi) -C(=NR 15 )_, (Vii) -C(R 37 a)(OR 37 c)_, (Viii) C(R 37 a)(SR 37 c)-, (iX) -C(R 37 a) (N(R 37 b)(R 37 c))-, (X) -C(R 37 a) (R 37 b)-o-, (xi) -~C(R 3 7 a)(R 37 b)-N(R 37 c)_, (xii) -C(R 37 a)(R 37 b )-N(O)(R 37 c)_, (xiii) -C(R 37 a)(R 3 7 b)-N (OH)-, (xiv) -~C(R 37 a) (R 3 7 b)S_ (xv) -C(R 37 a)(R 37 b)_S(O)_, (xvi) _C(R 37 a) (R 3 7 b)-S(O) 2 -, (xix) -C(R 37 a) (R 37 b)_C(= N R 1 5)_, (xx) -C(R 37 a) (OR 37 c)_C(=O)-, (xxi) -C(R 37 1) (SR 37 c)_C(=O)_, (xxii) -C(R 37 a) (OR 37 c)_C(=S)_, (xxiii) -C(R 1 7 a) (S R 37 c)_C(=S)_, (xxiv) -C(=O)-C(R 37 a)(OR 37 c)_, (xxv) -C(=O)-C(R 37 a) (SR 37 c)_, (xxvi) -C(=S)-C(R 37 a)(OR 37 c)_, (xxvii) -C(=S)-C(R 37 a) (SR 37 c)_, (xxviii) -C(R 37 a)(OR 37 c)_C(R 37 a)(OR 37 c)_, (xxix) -C(R 37 a)(S R 37 c)_C(R 37 a)(O R 37 c)-, (xxx) -C(R 37 a)(OR 37 c)_C(R 3 7 a)(SR 37 c)-, (xxxi) -C(R 37 a)(S R 37 c)_C(R 37 a) (SR 37 c)_, (xxxii) -CQ=O)-C(0O)-, (xxxiii) -C(=S)-C(=S)-, (xxxiv) -C(=O)-O-, (xxxv) -C(=O)-S-, (xxxix) -~C(=S)-N(R 37 a)_, (xA) -C(R 37 a)(R 3 7 b)_C&=O)-N(R 37 a)_, -9- WO 99/54299 PCT/US99/07945 (xliii) -C(R 37 a)(R 37 b)-C(=O)-S
-
, (xliv) -C(R 37 a)(R 37 b)-C(=S)-O-, (xlv) -C(R 37 a)(R 37 b)-C(=S)-S-, (xlvi) -C(R 37 a)(R 37 b)-N(R 37 b)-C(=O)-, (xivii) -C(R37a)(R37b)-N(R37b)-C(=S) - , (xlviii) -C(R37a)(R37b)-O-C(=O)-, (xlix) -C(R37a)(R37b)-S-C(=O) -, (1) -C(R37a)(R37b)-o-C(=S) -, (li) -C(R37a)(R37b)-S-C(=S)-, (lii) -C(R37a)(R37b)-N(R37b)-C(= O)-N(R37a)_, (liii) -C(R37a)(R37b)-N(R37b)-C(=S)-N(R37a) (liv) -C(R37a)(R37b)-N(R37b)-C(=O)-O - , (Iv) -C(R37a)(R37b)-N(R37b)-C(=O)-S - , (Ivi) -C(R37a)(R37b)-N(R37b)-C(=S)-O - , (Ivii) -C(R 37 a)(R 37 b)-N(R 37 b)-C(=S)-S -, (Iviii) -C(R 3 7 a)(R 37 b)-O-C(=O)-N(R 37 a), (lix) -C(R37a)(R37b)-S-C(=O)-N(R37a) , (Ix) -C(R 37 a)(R 37 b)-O-C(=S)-N(R 37 a)_, (Ixi) -C(R37a)(R37b)-S-C(=S)-N(R37a) - , (Ixii) -C(R37a)(R37b)-O-C(=O)-O - , (Ixiii) -C(R37a)(R37b)-S-C(=O)-O -, (lxiv) -C(R37a)(R37b)-o-C(=O)-S - , (lxv) -C(R 37 a)(R 37 b)-S-C(=O)-S - , (Ixvi) -C(R37a)(R37b)-o-C(=S)-O-, (Ixvii) -C(R37a)(R37b)-S-C(=S)-O -, (Ixviii) -C(R37a)(R37b)-o-C(=S)-S-, (Ixix) -C(R37a)(R37b)-S-C(=S)-S - or (lxx) -C(R 37 a)(R 37 b)-C(R 37 a)(OR 37 C);
R
14 is (i) hydrogen, (ii) C 1
-C
12 alkyl, (iii) haloalkyl, (iv) hydroxyalkyl, (v) thiol-substituted alkyl, (vi) R 37 cO-substituted alkyl, -10- WO 99/54299 PCT/US99/07945 (vii) R 37 cS-substituted alkyl, (viii) aminoalkyl, (ix) (R 37 c)NH-substituted alkyl, (x) (R 3 7 a)(R 37 c)N-susbstituted alkyl, (xi) R 37 aO-(O=)C-substituted alkyl, (xii) R 37 aS-(O=)C-substituted alkyl, (xiii) R 37 aO-(S=)C-substituted alkyl, (xiv) R 37 aS-(S=)C-substituted alkyl, (xv) (R 37 aO) 2 -P(=O)-substituted alkyl, (xvi) cyanoalkyl, (xvi) C 2
-C
12 alkenyl, (xviii) haloalkenyl, (xix) C 2
-C
12 alkynyl, (xx) cycloalkyl, (xxi) (cycloalkyl)alkyl, (xxii) (cycloalkyl)alkenyl, (xxiii) (cycloalkyl)alkynyl, (xxiv) cycloalkenyl, (xxv) (cycloalkenyl)alkyl, (xxvi) (cycloalkenyl)alkenyl, (xxvii) (cycloalkenyl)alkynyl, (xxviii) aryl, (xxix) (aryl)alkyl, (xxx) (aryl)alkenyl, (xxxi) (aryl)alkynyl, (xxxii) heterocyclic, (xxxiii) (heterocyclic)alkyl, (xxxiv) (heterocyclic)alkenyl or (xxxv) (heterocyclic)alkynyl, with the proviso that R 14 is other than hydrogen when Z is -C(R 37a)(R37b)-N(R37b)-C(=O)-o - , -C(R37a) (R37b)-N(R37b)-C(=S)-O - ,
-C(R
37 a)(R 37 b)-N(R 37 b)
-
C(=O)
-S - , -C(R 37 a)(R 37 b)
-
N(R
37 b)
-
C(=S)
-S -, -C(R37a)(R37b)-o-C(=O)-O - , -C(R37a) (R37b)-O-C(= S)-O -, -C(R37a)(R37b)-S-C(=O)-O -, -C(R37a)(R37b)-S-C(=S)-O - , -C(R37a)(R37b)-o-C(=O)-S
-
, -C(R37a) (R37b)-O-C(=S)-S - , -11- WO 99/54299 PCT/US99/07945 -C(R37a)(R37b)-S-C(=O)-S - or -C(R37a)(R37b)-S-C(=S)-S-;
SR
3 7a ,
R
37 b, R 47 , and R 48 at each occurrence are independently selected from the group consisting of (i) hydrogen, (ii) C1-C12 alkyl, (iii) haloalkyl, (iv) hydroxyalkyl, (v) alkoxyalkyl, (vi) C2-C12 alkenyl, (vii) haloalkenyl, (viii) C2-012 alkynyl, (ix) cycloalkyl, (x) (cycloalkyl)alkyl, (xi) (cycloalkyl)alkenyl, (xii) (cycloalkyl)alkynyl, (xiii) cycloalkenyl, (xiv) (cycloalkenyl)alkyl, (xv) (cycloalkenyl) alkenyl, (xvi) (cycloalkenyl)alkynyl, (xvii) aryl, (xviii) (aryl)alkyl, (xix) (aryl)alkenyl, (xx) (aryl)alkynyl, (xxi) heterocyclic, (xxii) (heterocyclic)alkyl, (xxiii) (heterocyclic)alkenyl and (xxiv) (heterocyclic)alkynyl;
R
37 c at each occurrence is independently selected from the group consisting of (i) hydrogen, (ii) Cl-C12 alkyl, (iii) haloalkyl, (iv) C2-C12 alkenyl, (v) haloalkenyl, (vi) C2-012 alkynyl, (vii) cycloalkyl, (viii) (cycloalkyl)alkyl, (ix) (cycloalkyl)alkenyl, (x) (cycloalkyl)alkynyl, (xi) cycloalkenyl, (xii) (cycloalkenyl)alkyl, (xiii) (cycloalkenyl)alkenyl, (xiv) (cycloalkenyl)alkynyl, (xv) aryl, (xvi) (aryl)alkyl, -12- WO 99/54299 PCT/US99/07945 (xvii) (aryl)alkenyl, (xviii) (aryl)alkynyl, (xix) heterocyclic, (xx) (heterocyclic)alkyl, (xxi) (heterocyclic)alkenyl, (xxii) (heterocyclic)alkynyl, (xxiii) -C(=O)-R 14 , (xxiv) -C(=S)-R 4 , (xxv) -S(O) 2
-R
14 and (xxvi) hydroxyalkyl; or when Z is -C(R 37 a)(R 37 b)-N(R 37 c)-, then N(R 37 c) and R 1 4 when taken together are an azido group; or when Z is -C(R 37 a)(R 37 b)-N(O)(R 37 c)
-
, then N(O)(R 37c ) and R 14 when taken together are an N-oxidized 3-7 membered heterocyclic ring having at least one N-oxidized ring nitrogen atom; or when Z is -C(R 37 a)(OR 37 c)
-
, -C(R 37 a)(SR 3 7 c) - or -C(R3 7 a)(N(R 37 b)(R 37 c))
-
, then R 3 7 a, R 14 and the carbon atom to which they are bonded when taken together form a cyclopentyl, cyclopentenyl, cyclohexyl or cyclohexenyl ring;
R
15 is selected from the group consisting of (i) hydrogen, (ii) hydroxy, (iii) amino, (iv) C1-C12 alkyl, (v) haloalkyl, (vi) C2-012 alkenyl, (vii) haloalkenyl, (viii) cycloalkyl, (ix) (cycloalkyl)alkyl, (x) (cycloalkyl)alkenyl, (xi) cycloalkenyl, (xii) (cycloalkenyl)alkyl, (xiii) (cycloalkenyl)alkenyl, (xiv) aryl, (xv) (aryl)alkyl, (xvi) (aryl)alkenyl, (xvii) heterocyclic, (xviii) (heterocyclic)alkyl and (xix) (heterocyclic)alkenyl; -13- WO 99/54299 PCT/US99/07945 or R 3 and R 4 taken together, with the atom to which they are attached, form a carbocyclic or heterocyclic ring having from 3 to 8 ring atoms;
R
5 is selected from the group consisting of (a) hydrogen, (b) -CH(R 38
)
2 , (c) -O-R 40 , (d) C2-C4 alkynyl, (e) cyclopropyl, (f) cyclobutyl, (g) -C(=Q 1
)-R
17 , and (h) -N(R 1 9
)
2 wherein Q 1 is O, S, or N(R 18 );
R
17 and R 18 are independently selected, at each occurrence, from the group consisting of hydrogen, methyl, and ethyl;
R'
9 , R 38 , and R 4 0 are independently selected, at each occurrence, from the group consisting of (i) hydrogen, (ii) C1-C12 alkyl, (iii) haloalkyl, (iv) C2-C12 alkenyl, (v) haloalkenyl, (vi) cycloalkyl, (vii) (cycloalkyl)alkyl, (viii) (cycloalkyl)alkenyl, (ix) cycloalkenyl, (x) (cycloalkenyl)alkyl, (xi) (cycloalkenyl)alkenyl, (xii) aryl, (xiii) (aryl)alkyl, (xiv) (aryl)alkenyl, (xv) heterocyclic, (xvi) (heterocyclic)alkyl and (xvii) (heterocyclic)alkenyl; Y is selected from the group consisting of (a) hydrogen, (b) C1-C5 alkyl, (c) C1-C5 haloalkyl, (d) C2-C5 alkenyl, (e) C2-05 haloalkenyl, (f) C2-C5 alkynyl, (g) C3-C5 cycloalkyl, (h) C3-05 cycloalkyl-C0-to-C 3 -alkyl, (i) C5 cycloalkenyl, (j) C5 cycloalkenyl-Cl-to
C
3 -alkyl, (k) C5 cycloalkenyl-C 2 -to-C 3 -alkenyl, (I) -(CHR 39 )nOR 20 , (m) -CH(OR 20
)
CH
2
(OR
20 ), (n) -(CHR 39 )nSR 21 , (0) -(CHR 39 )nCN, (p) -(CHR 39 )nN 3 , (q) phenyl, -14- WO 99/54299 PCT/US99/07945 (r) halo-substituted phenyl, (s) -(CHR 3 9 )nC (=Q2)R 22 , (t) -(CHR 39 )n (=Q 3 ), (u) -N(O)=CHCH 3 , (v) -(CHR 39 )nNR 23
R
24 , (w) halo and (x) a heterocyclic ring having from 3 to 6 ring atoms; wherein n is 0, 1, or 2; Q 2 is O, S, NR 25 , or CHR 26 ; and Q3 is NR 41 , or CHR 42
R
20 at each occurrence is independently (i) hydrogen, (ii) methyl, (iii) ethyl, (iv) n-propyl, (v) isopropyl, (vi) C1-C3 haloalkyl, (vii) vinyl, (viii) propenyl, (ix) isopropenyl, (x) allyl, (xi) C2-C3 haloalkenyl, (xii) amino, (xiii) -NHCH 3 , (xiv) -N(CH 3
)
2 , (xv) -NHCH 2
CH
3 , (xvi) -N(CH 3
)(CH
2
CH
3 ), (xvii) -N(CH 2
CH
3
)
2 or (xviii) -N(=CH 2 );
R
21 is hydrogen, (ii) methyl, (iii) ethyl, (iv) n-propyl, (v) isopropyl, (vi) C1-C3 haloalkyl, (vii) vinyl, (viii) propenyl, (ix) isopropenyl, (x) allyl or (xi) C2-C3 haloalkenyl;
R
2 2 is (i) hydrogen, (ii) methyl, (iii) ethyl, (iv) n-propyl, (v) isopropyl, (vi) hydroxy, (vii) thiol, (viii) methoxy, (ix) ethoxy, (x) n-propoxy, (xi) isopropoxy, (xii) cyclopropyloxy, (xiii) methylthio, (xiv) ethylthio, (xv) n-propylthio, (xvi) isopropylthio, (xvii) cyclopropylthio, (xviii) vinyl, (xix) propenyl, (xx) isopropenyl, (xxi) allyl, (xxii) -N(R 2 8 a)(R 2 8 b), (xxiii) -CH 2
R
29 , (xxiv) aminomethyl, (xxv) hydroxymethyl, -15- WO 99/54299 PCT/US99/07945 (xxvi) thiolmethyl, (xxvii) -NHNH 2 , (xxviii) -N(CH 3
)NH
2 or (xxix) -NHNH(CH 3 );
R
2 3 and R 39 are independently hydrogen or methyl;
R
41 and R 42 are independently hydrogen, methyl, or ethyl;
R
24 is selected from the group consisting of (i) hydrogen, (ii) Cl-C4 alkyl, (iii) C 2
-C
4 alkenyl, (iv) C2-C4 alkynyl, (v) cyclopropyl, (vi) -C(=Q 4
)-R
30 , (v) -OR 31 , and (vi) -N(R 32
)
2 , wherein Q 4 is O, S, or N(R33);
R
25 is hydrogen, hydroxy, methyl, ethyl, amino, -CN, or -NO 2 ;
R
26 group is hydrogen, methyl or ethyl;
R
28 a hydrogen, hydroxy, methyl, ethyl, amino, -NHCH 3 , -N(CH 3
)
2 , methoxy, ethoxy, or -CN;
R
28 b is hydrogen, methyl or ethyl; or R 28 a ,
R
28 b and the nitrogen to which they are bonded taken together represent azetidinyl;
R
29 group is hydrogen, hydroxy, thiol, methyl, ethyl, amino, methoxy, ethoxy, methylthio, ethylthio, methylamino or ethylamino;
R
30 group is hydrogen, methyl, ethyl, -OR 34 , -SR 34 , -N(R 35
)
2 , -NHOH,
-NHNH
2 , -N(CH 3
)NH
2 , or -N(CH 2
CH
3
)NH
2 ;
R
31 and R 32 substituents, at each occurrence, are independently hydrogen, methyl or ethyl;
R
33 group is hydrogen, hydroxy, methyl, ethyl, amino, -CN, or -NO 2 ; -16- WO 99/54299 PCT/US99/07945
R
34 group is methyl or ethyl;
R
35 group is independently hydrogen, methyl or ethyl; with the proviso that when Q2 is CHR 26 then R 22 is selected from the group consisting of hydrogen, -CH 3 , -C 2
H
5 , -C 3
H
7 , -OCH 3 , -SCH 3 , -O-0 2
H
5 , and
-S-C
2
H
5 , and with the proviso that when R 3 and R 4 are each hydrogen, then Y is other than hydrogen;
R
6 and R 7 are independently selected from the group consisting of (a) hydrogen, (b) C 1
-C
12 alkyl, (c) C 2
-C
12 alkenyl, (d) cycloalkyl, (e) (cycloalkyl)alkyl, (f) (cycloalkyl)alkenyl, (g) cycloalkenyl, (h) (cycloalkenyl)alkyl, (i) (cycloalkenyl)alkenyl, (j) aryl, (k) (aryl)alkyl, (I) (aryl)alkenyl, (m) heterocyclic, (n) (heterocyclic)alkyl and (0) (heterocyclic)alkenyl; and
R
8 , R 9 , and R 1 0 are independently selected from the group consisting of (a) hydrogen, (b) Cl-C 6 alkyl, (c) C 2
-C
6 alkenyl, (d) C 3
-C
6 cycloalkyl, (e) C 3
-C
6 cycloalkenyl, and (f) fluorine, with the proviso that the total number of atoms, other than hydrogen, in each of R 8 , R 9 , and R 1 0 , is 6 atoms or less. -17- WO 99/54299 PCT/US99/07945 Preferred compounds of the invention are compounds having the relative sterochemistry depicted by Formula II:
R
9
R
8 Rio R 6 R2-X,'. ' R 0 / R 6 R4 R 7\ "5 R R
R
3 II or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein R 1 , R 2 ,
R
3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 0 , X and Y are as defined above and wherein R 3 and
R
4 are not both the same. Other preferred compounds of the invention are compounds having the relative sterochemistry depicted by Formula III:
SR
9 R8 6 R2-XN 1
R
3 III -18- WO 99/54299 PCT/US99/07945 or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein R 1 , R 2 ,
R
3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 o, X and Y are as defined above and wherein R 3 and
R
4 are not both the same. Other preferred compounds of the invention are compounds having Formula I or II or III or a salt, ester or prodrug thereof wherein R 1 is defined as above;
-X-R
2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2
-NH-SO
2 - or R 2
-SO
2
-NH
wherein R 2 is 01-03 loweralkyl, halo C 1
-C
3 loweralkyl, C2-03 alkenyl or halo 02-03 alkenyl or -X-R 2 is O -- y2 Y 2 wherein Y' is -CH 2 -, -0-, -S- or -NH- and Y 2 is -C(=O)- or -C(Raa)( Rbb)- wherein Raa and Rbb are independently selected from the group consisting of hydrogen, C1-03 loweralkyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, aminomethyl, 1-aminoethyl, 2-aminoethyl, thiolmethyl, 1-thiolethyl, 2-thiolethyl, methoxymethyl, N-methylaminomethyl and methylthiomethyl;
R
3 and R 4 are independently selected from hydrogen, heterocyclic and
-Z-R
14 wherein Z and R14 are defined as above and wherein one of R 3 and R 4 is other than hydrogen;
R
5 is hydrogen or loweralkyl;
R
6 and R 7 are independently hydrogen or loweralkyl;
R
8 and R 9 are independently hydrogen, fluoro or loweralkyl; -19- WO 99/54299 PCT/US99/07945
R
1 0 is hydrogen, fluoro or loweralkyl; and Y is C 2
-C
5 alkenyl, C 2
.C
5 haloalkenyl, -C(=Q 2
)R
22 ,
-N(=Q
3 ), -N(O)=CHCH 3 , -NR 2 3
R
24 or a heterocyclic ring having from 3 to 6 ring atoms, wherein R 2 2 , R 23 , R 24 , Q 2 and Q 3 are defined as above. More preferred compounds of the invention are compounds having Formula I or II or III or a salt, ester or prodrug thereof wherein R' is defined as above;
-X-R
2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2
-NH-SO
2 - or R 2
-SO
2
-NH
wherein R 2 is 0C 1
-C
3 Ioweralkyl, halo C 1
-C
3 loweralkyl, 0 2
-C
3 alkenyl or halo C 2 -C3 alkenyl or -X-R 2 is O
Y
1 2 wherein Y' is -CH 2 - and Y 2 is -C(=O)- or -C(Raa)( Rbb) - wherein Raa and Rbb are independently selected from the group consisting of hydrogen, Cl-C 3 loweralkyl, hydroxymethyl, 1-hydroxyethyl and 2-hydroxyethyl;
R
3 and R 4 are independently selected from hydrogen, heterocyclic and -Z-R 14 wherein Z and R 14 are defined as above and wherein one of R 3 and R 4 is other than hydrogen;
R
5 is hydrogen or loweralkyl;
R
6 and R 7 are independently hydrogen or loweralkyl;
R
8 and R 9 are independently hydrogen or loweralkyl;
R
1 0 is hydrogen or loweralkyl; and -20- WO 99/54299 PCT/US99/07945 Y is 02-05 alkenyl, 0 2
-C
5 haloalkenyl, -C(=Q 2
)R
22 ,
-N(=Q
3 ), -N(O)=CHCH 3 or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds, wherein R 2 2 , Q2 and Q 3 are defined as above. Even more preferred compounds of the invention are compounds having Formula I or II or III or a salt, ester or prodrug thereof wherein R 1 is defined as above;
-X-R
2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2
-NH-SO
2 - or R 2
-SO
2
-NH
wherein R 2 is 01C-03 loweralkyl, halo C1-C3 loweralkyl, 02-03 alkenyl or halo C 1
-C
3 alkenyl or -X-R 2 is 0
Y
1 - 2 Y wherein Y 1 is -OH 2 - and Y 2 is -C(=O)- or -C(Raa)( Rbb) - wherein Raa and Rbb are independently selected from the group consisting of hydrogen, C1-C3 loweralkyl, hydroxymethyl, 1-hydroxyethyl and 2-hydroxyethyl;
R
3 and R 4 are independently selected from hydrogen, heterocyclic and
-Z-R
14 wherein Z and R 14 are defined as above and wherein one of R 3 and R 4 is other than hydrogen;
R
5 is hydrogen or loweralkyl;
R
6 and R 7 are independently hydrogen or loweralkyl;
R
8 and R 9 are independently hydrogen or loweralkyl;
R
10 is hydrogen or loweralkyl; and -21- WO 99/54299 PCT/US99/07945 Y is C 2
-C
5 alkenyl, C2.C 5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. More highly preferred compounds of the invention are compounds having Formula I or II or III or a salt, ester or prodrug thereof wherein R 1 is -CO 2 H;
-X-R
2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2
-NH-SO
2 - or R 2
-SO
2 -NH- wherein R 2 is
C
1
-C
3 loweralkyl or halo- Cl-C 3 loweralkyl;
R
3 and R 4 are independently selected from hydrogen, heterocyclic and -Z-R 1 4 wherein Z and R 14 are defined as above and wherein one of R 3 and R 4 is other than hydrogen;
R
5 is hydrogen or loweralkyl;
R
6 and R 7 are hydrogen independently hydrogen or loweralkyl;
R
8 and R 9 are hydrogen independently hydrogen or loweralkyl;
R
1 0 is hydrogen or loweralkyl; and Y is C 2
-C
5 alkenyl, C 2
-C
5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. Even more highly preferred compounds of the invention are compounds having Formula I or II or III or a salt, ester or prodrug thereof wherein R, is CO 2 H;
-X-R
2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2
-NH-SO
2 - or R 2
-SO
2 -NH- wherein R 2 is Cl-C 3 loweralkyl or halo- Cl-C 3 loweralkyl;
R
4 is hydrogen or loweralkyl and R 3 is heterocyclic or -Z-R 14 wherein Z and R 14 are defined as above;
R
5 is hydrogen; -22- WO 99/54299 PCT/US99/07945
R
6 and R 7 are hydrogen;
R
8 and R 9 are hydrogen;
R
1 0 is hydrogen; and Y is C 2
-C
5 alkenyl, C2-C5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. Other even more highly preferred compounds of the invention are compounds having Formula I or II or III or a salt, ester or prodrug thereof wherein
R
1 is -CO 2 H;
-X-R
2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2
-NH-SO
2 - or R 2
-SO
2 -NH- wherein R 2 is
C-C
3 loweralkyl or halo Cl-C3 loweralkyl;
R
4 is hydrogen or loweralkyl and R 3 is (a) heterocyclic, (b) alkyl, (b) cycloalkyl, (d) cycloalkylalkyl, (e) alkenyl, (f) alkynyl, (g) -C(=O)-R 1 4 (h) -C(R 37 a)(OR 37 c)-R1 4 or (i) -C(R3 7 a)(R 37 b)-N(O)(R 37 c)R1 4 wherein R 14 is (i) alkyl, (ii) cycloalkyl, (iii) cycloalkylalkyl, (iv) alkenyl, (v) haloalkyl, (vi) haloalkenyl, (vii) aryl, (viii) arylalkyl, (ix) heterocyclic, (x) (heterocyclic)alkyl, (xi) hydroxyalkyl, (xii) alkoxyalkyl, (xiii) cyanoalkyl, (xiv) (R 37 aO)-(O=)C-substituted alkyl or (xv) (R3 7 aO) 2 -P(=O)-substituted alkyl;
R
3 7a and R 37 b are independently selected from the group consisting of (i) hydrogen, (ii) loweralkyl and (iii) loweralkenyl; and
R
37 c is hydrogen, (ii) loweralkyl or (iii) loweralkenyl; -23- WO 99/54299 PCT/US99/07945
R
5 is hydrogen;
R
6 and R 7 are hydrogen;
R
8 and R 9 are hydrogen;
R
1 0 is hydrogen; and Y is C 2
-C
5 alkenyl, C 2
-C
5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. Most highly preferred compounds of the invention are compounds having Formula I or II or III or a salt, ester or prodrug thereof wherein R' is -CO 2 H;
-X-R
2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2
-NH-SO
2 - or R 2
-SO
2 -NH- wherein R 2 is Cl-C 3 loweralkyl or halo C 1
-C
3 loweralkyl;
R
4 is hydrogen and R 3 is (a) heterocyclic, (b) alkyl or (c) -C(R 37 a)(OR 37 c) R 14 wherein R 14 is (i) alkyl, (ii) cycloalkyl, (iii) cycloalkylalkyl, (iv) alkenyl, (v) haloalkyl, (vi) haloalkenyl, (vii) aryl, (viii) arylalkyl, (ix) heterocyclic, (x) (heterocyclic)alkyl, (xi) hydroxyalkyl, (xii) alkoxyalkyl, (xiii) cyanoalkyl, (xiv) (R 37 aO)-(O=)C-substituted alkyl or (xv) (R 37 aO) 2 -P(=O)-substituted alkyl;
R
37 a and R 37 b are independently selected from the group consisting of (i) hydrogen, (ii) loweralkyl and (iii) loweralkenyl; and
R
37 c is hydrogen, (ii) C 1
-C
3 loweralkyl or (iii) allyl;
R
5 is hydrogen; -24- WO 99/54299 PCT/US99/07945
R
6 and R 7 are hydrogen;
R
8 and R 9 are hydrogen;
R
1 0 is hydrogen; and Y is C2-C5 alkenyl, C 2
.C
5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. Other most highly preferred compounds of the invention are compounds having Formula I or II or III or a salt, ester or prodrug thereof wherein R 1 is CO 2 H1;
-X-R
2 is R 2 -C(=O)-NH- or R 2
-SO
2 -NH- wherein R 2 is C 1
-C
3 loweralkyl or halo C C3 loweralkyl;
R
4 is hydrogen and R 3 is (a) heterocyclic, (b) alkyl or (c) -C(R 37 a)(OR 37 c)
R
14 wherein R 14 is (i) loweralkyl, (ii) loweralkenyl, (iii) hydroxy-substituted loweralkyl or (iv) alkoxy-substituted loweralkyl;
R
3 7a is (i) hydrogen, (ii) loweralkyl or (iii) loweralkenyl; and
R
3 7 c is (i) hydrogen, (ii) C1-C3 loweralkyl or (iii) allyl;
R
5 is hydrogen;
R
6 and R 7 are hydrogen;
R
8 and R 9 are hydrogen;
R
1 0 is hydrogen; and -25- WO 99/54299 PCT/US99/07945 Y is C 2
-C
5 alkenyl, C 2 .- C 5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. Other most highly preferred compounds of the invention are compounds having Formula I or II or III or a salt, ester or prodrug thereof wherein R 1 is CO 2 H;
-X-R
2 is R 2 -C(=O)-NH- or R 2
-SO
2 -NH- wherein R 2 iS 0 1 -0 3 loweralkyl or halo C 1 C 3 loweralkyl;
R
4 is hydrogen and R 3 is -C(R 37 a)(OR 3 7 c)-R1 4 wherein R 1 4 is loweralkyl or loweralkenyl;
R
37 a is loweralkyl or loweralkenyl; and
R
37 c is hydrogen, C 1
-C
3 loweralkyl or allyl;
R
5 is hydrogen;
R
6 and R 7 are hydrogen;
R
8 and R 9 are hydrogen;
R
10 is hydrogen; and Y is C 2
-C
5 alkenyl, C 2
-C
5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. Preferred substituents R 1 include -CO 2 H or esters or prodrugs thereof. Preferred esters include C2-C6 loweralkyl esters or substituted or unsubstituted -26- WO 99/54299 PCT/US99/07945 benzyl esters. Preferred substituents R 1 also include -S(O) 2
NHC(=O)R
1 wherein R 11 is defined as above. Most highly preferred substituents R 1 include -CO 2 H or esters or prodrugs thereof. Most highly preferred esters include C2-C6 loweralkyl esters or substituted or unsubstituted benzyl esters. Preferred substituents -X-R 2 include R 2 -C(=O)-NH-, R 2 -NH-C(=O)-,
R
2
-NH-SO
2 - or R 2
-SO
2 -NH- wherein R 2 is 01-03 loweralkyl, halo C1-C3 loweralkyl, 02-03 alkenyl or halo C2-C3 alkenyl or -X-R 2 is Y O
/-
Y wherein Y' is -CH 2 -, -0-, -S- or -NH- and Y 2 is -C(=O)- or -C(Raa)( Rbb)- wherein Ra " and Rbb are independently selected from the group consisting of hydrogen, C-C3 loweralkyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, aminomethyl, 1-aminoethyl, 2-aminoethyl, thiolmethyl, 1-thiolethyl, 2-thiolethyl, methoxymethyl, N-methylaminomethyl and methylthiomethyl. More preferred substituents -X-R 2 include R 2 -C(=O)-NH-, R 2 -NH-C(=O)-,
R
2
-NH-SO
2 - or R 2
-SO
2 -NH- wherein R 2 is 01-03 loweralkyl, halo C01-C3 loweralkyl, 02-03 alkenyl or halo C2-C3 alkenyl or -X-R 2 is 0
-
wherein Y' is -OH 2 - and Y 2 is -C(=O)- or -C(Raa)( Rbb) - wherein Raa and Rbb are independently selected from the group consisting of hydrogen, -27- WO 99/54299 PCT/US99/07945 Cl-C 3 loweralkyl, hydroxymethyl, 1-hydroxyethyl and 2-hydroxyethyl. Even more preferred substituents -X-R 2 include R 2 -C(=O)-NH-, R 2 -NH-C(=O)-,
R
2
-NH-SO
2 - or R 2
-SO
2 -NH- wherein R 2 is C 1
-C
3 loweralkyl, halo C 1
-C
3 loweralkyl,
C
2
-C
3 alkenyl or halo C 2
-C
3 alkenyl. More highly preferred substituents -X-R 2 include R 2 -C(=O)-NH-, R 2 -NH-C(=O)-,
R
2
-NH-SO
2 - or R 2
-SO
2 -NH- wherein R 2 is C 1
-C
3 Ioweralkyl or halo- C 1
-C
3 loweralkyl. Even more highly preferred substituents-X-R 2 include R 2 -C(=O)-NH-,
R
2 -NH-C(=O)-, R 2
-NH-SO
2 - or R 2
-SO
2 -NH- wherein R 2 is C 1
-C
2 loweralkyl or halo
C
1
-C
2 Ioweralkyl, and especially, CH 3 -C(=O)-NH-, CF 3 -C(=O)-NH-, CH 3
-SO
2
-NH
or CF 3 -SO2-NH-. Preferred substituents R 3 and R 4 are independently selected from the group consisting of hydrogen, heterocyclic and -Z-R 14 wherein Z and R 14 are defined as most broadly defined previously herein and wherein one of R 3 and R 4 is other than hydrogen. More highly preferred, substituent R 4 is hydrogen or loweralkyl and R 3 includes heterocyclic or -Z-R 1 4 wherein Z and R 14 are defined as most broadly defined previously herein. Even more highly preferred, substituent R 4 is hydrogen or loweralkyl and
R
3 includes (a) heterocyclic, (b) alkyl, (c) cycloalkyl, (d) cycloalkylalkyl, (e) alkenyl, (f) alkynyl, (g) -C(=O)-R 14 , (h) -C(R 37 a)(OR 37 c)-R 1 4 or (i) -C(R 37 a)(R 37 b)-N(O)(R 37 c)R 14 wherein R 14 is (i) alkyl, (ii) cycloalkyl, (iii) cycloalkylalkyl, (iv) alkenyl, (v) haloalkyl, (vi) haloalkenyl, (vii) aryl, (viii) arylalkyl, (ix) heterocyclic, -28- WO 99/54299 PCT/US99/07945 (x) (heterocyclic)alkyl, (xi) hydroxyalkyl, (xii) alkoxyalkyl, (xiii) cyanoalkyl, (xiv) (R 3 7 aO)-(O=)C-substituted alkyl or (xv) (R 37 aO) 2 -P(=O)-substituted alkyl;
R
37a and R 3 7 b are independently selected from the group consisting of (i) hydrogen, (ii) loweralkyl and (iii) loweralkenyl; and
R
37 c is (i) hydrogen, (ii) loweralkyl or (iii) loweralkenyl. Most highly preferred, substituent R 4 is hydrogen and R 3 includes (a) heterocyclic, (b) alkyl or (c) -C(R 37 a)(OR 37 c)-R 14 wherein R 14 is (i) alkyl, (ii) cycloalkyl, (iii) cycloalkylalkyl, (iv) alkenyl, (v) haloalkyl, (vi) haloalkenyl, (vii) aryl, (viii) arylalkyl, (ix) heterocyclic, (x) (heterocyclic)alkyl, (xi) hydroxyalkyl, (xii) alkoxyalkyl, (xiii) cyanoalkyl, (xiv) (R 37 aO)-(O=)C-substituted alkyl or (xv) (R 37 aO) 2 -P(=O)-substituted alkyl;
R
37a and R 37 b are independently selected from the group consisting of (i) hydrogen, (ii) loweralkyl and (iii) loweralkenyl; and
R
37 c is (i) hydrogen, (ii) C1-C3 loweralkyl or (iii) allyl. Also most highly preferred, substituent R 4 is hydrogen and R 3 includes (a) heterocyclic, (b) alkyl or (c) -C(R 37 a)(OR 37 c)-R1 4 wherein R 1 4 is (i) loweralkyl, (ii) loweralkenyl, (iii) hydroxy-substituted loweralkyl or (iv) alkoxy-substituted loweralkyl;
R
3 7a is (i) hydrogen, (ii) loweralkyl or (iii) loweralkenyl; and
R
37 c is. (i) hydrogen, (ii) C1-C3 loweralkyl or (iii) allyl. -29- WO 99/54299 PCT/US99/07945 Also most highly preferred, substituent R 4 is hydrogen and R 3 includes
-C(R
37 a)(OR 37 c)-R1 4 wherein R 14 is loweralkyl or loweralkenyl;
R
37a is loweralkyl or loweralkenyl; and
R
3 7 c is hydrogen, C1-C3 loweralkyl or allyl, and especially, wherein R 37 c is hydrogen or methyl. Preferred substituents R 5 include hydrogen or loweralkyl. Most highly preferred, R 5 is hydrogen. Preferred substituents R 6 and R 7 include independently hydrogen and loweralkyl. Most highly preferred, R 6 and R 7 are hydrogen. Preferred substituents R 8 , R 9 and R 1 0 incude independently hydrogen, fluoro and loweralkyl. Most highly preferred, R 8 , R 9 and R 10 are hydrogen. Preferred substituent Y includes C2-C5 alkenyl, C2.-C5 haloalkenyl,
-C(=Q
2
)R
22 , -N(=Q 3 ), -N(O)=CHCH 3 , -NR 23
R
24 or a heterocyclic ring having from 3 to 6 ring atoms, wherein R 22 , R 2 3 , R 24 , Q 2 and Q 3 are defined as above. More preferred substituent Y includes C2-C5 alkenyl, C2-.C5 haloalkenyl, -C(=Q 2
)R
22 , -N(=Q 3 ), -N(O)=CHCH 3 or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds, wherein R 22
Q
2 and Q 3 are defined as above. Even more preferred substituent Y includes C2-C5 alkenyl, C2-C5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. Representative alkenyl and haloalkenyl substituents Y include:
-CH=CH
2 , -CH=CHF, -CH=CH-CH 3 , -CH=CH-CF 3 , -CH=CHCI, -CH=CHBr,
-CH=CF
2 , -CH=CF(CH 3 ), -CH=CF(CF 3 ), -CH=CFCI, -CH=CFBr, -CH=C(CH 3
)
2 , -30- WO 99/54299 PCTIUS99/07945
-CH=C(CH
3 )(0F 3 ), -CH=CCI(0H 3 ), -CH=CBr(CH 3 ), -CH=C(CF 3
)
2 , -CH=CCI(CF 3 ), -CH=CBr(CF 3 ), -CH=CCI 2 , -CH=CCIBr, -CF=0H 2 , -CF=CHF, -CF=CH-CH 3 ,
-CF=CH-CF
3 , -CF=CHCI, -CF=CHBr, -CF=CF 2 , -CF=CF(CH 3 ), -CF=CF(CF 3 ), -CF=CFCI, -CF=CFBr, .- CF=C(CH 3
)
2 , -CF=C(CH 3
)(CF
3 ), -CF=CCI(CH 3 ), -CF=CBr(CH 3 ), -CF=C(CF 3
)
2 , -CF=CCI(CF 3 ), -CF=CBr(CF 3 ), -CF=CC 2 , -CF=CCIBr, -C(CH 3
)=CH
2 , -C(CH 3 )=CHF, -C(CH 3
)=CH-CH
3 , -C(CH 3
)=CH-CF
3 ,
-C(CH
3 )=CHCI, -C(CH 3 )=CHBr,-C(CH 3
)=CF
2 , -C(CH 3
)=CF(CH
3 ), -C(0H 3
)=CF(CF
3 ), -C(CH 3 )=CFCt,
-C(CH
3 )=CFBr, -C(CH 3
)=C(CH
3
)
2 , -C(CH 3
)=C(CH
3 )(CFA) -C(CH 3
)=CCI(CH
3 ),
-C(CH
3 )=CBr(CH 3 ), -C(CH 3
)=C(CF
3
)
2 , -C(CH 3 )=C01(CF 3 ), -C(CH 3 )=CBr(CF 3 ),
-C(CH
3
)=CCI
2 , -C(CH 3 )=CCIBr,
-C(CF
3
)=CH
2 , -C(CF 3 )=CHF, -C(CF 3
)=CH-CH
3 , -C(CF 3
)=CH-CF
3 , -C(CF 3 )=CHCI,
-C(CF
3 )=CHBr, -C(CF 3
)=CF
2 , -C(CF 3
)=CF(CH
3 ), -C(CF 3
)=CF(CF
3 ), C(CF 3 )=CFCI, -C(CF 3 )=CFBr, -C(0F 3
)=C(CH
3
)
2 , -C(CF 3
)=C(CH
3
)(CF
3 ), C(CF 3 )=CCI(0H 3 ),
-C(CF
3 )=CBr(0H 3 ), -C(CF 3
)=C(CF
3
)
2 , -C(CF 3
)=CCI(CF
3 ), -C(CF 3 )=CBr(CF 3 ),
-C(CF
3 )=CC1 2 , -C(CF 3 )=CCIBr,
-CCI=CH
2 , -CCI=CHF, -CCI=CH-CH 3 , -CCI=CH-CF 3 , -CCI=CHCI, -CCI=CHBr,
-CCI=CF
2 , -CCI=CF(CH 3 ), -CCI=CF(CF 3 ), -CCI=CFCI, -CCI=CFBr, -CCI=C(CH 3
)
2 ,
-CCI=C(CH
3
)(CF
3 ), -CCI=CCI(CH 3 ), -CCI=CBr(CH 3 ), -CCI=C(0F 3
)
2 ,
-CCI=CCI(CF
3 ), -CCI=CBr(CFA) -CCI=CC1 2 , -CCI=CCIBr, -31- WO 99/54299 PCT/US99/07945
-CH=CH-CH
2
CH
3 , -CH=CF-CH 2
CH
3 , -CF=CH-CH 2
CH
3 , -CF=CF-CH 2
CH
3 ,
-CH=C(CH
3
)(CH
2
CH
3 ), -CF=C(CH 3
)(CH
2
CH
3 ), -CH=CCI(CH 2
CH
3 ),
-CF=CCI(CH
2
CH
3 ), -C(CH 3
)=CH-CH
2
CH
3 , -C(CH 3
)=CF-CH
2
CH
3 ,
-CCI=CH-CH
2
CH
3 , -CCI=CF-CH 2
CH
3 , -C(CH 2
CH
3
)=CH
2 , -C(CH 2
CH
3 )=CHF,
-C(CH
2
CH
3
)=CF
2 , -C(CH 2
CH
3
)=CH-CH
3 , -C(CH 2
CH
3
)=CF-CH
3 ,
-C(CH
2
CH
3 )=CH-CI, -C(CH 2
CH
3 )=CFCI. Representative Y substituents which are heterocyclic rings having 5 ring atoms and also containing one or two double bonds include: furanyl, dihydrofuranyl, didehydrodioxolanyl, dithiolyl, imidazolyl, imidazolinyl, isothiazolyl, isothiazolinyl, isoxazolyl, isoxazolinyl, oxadiazolyl, oxadiazolinyl, oxathiolyl, oxazolyl, oxazolinyl, pyrazolyl, pyrazolinyl, pyrrolyl, dihydropyrrolyl, tetrazolyl, tetrazolinyl, thiadiazolyl, thiadiazolinyl, thiazolyl, thiazolinyl, thienyl, dihydrothienyl, triazolyl, triazolinyl. More highly preferred substituents Y include cis-propenyl, trans-propenyl, isobutenyl, cis-2-chlorovinyl, vinyl, 2,2-difluorovinyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isoxazolyl. Most highly preferred substituents Y include cis-propenyl, cis-2-chlorovinyl, vinyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isoxazolyl. Preferred compounds of the invention include compounds selected from the group consisting of: -32- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R, 1'R)-2-(1 -Acetamido-2-ethyl-2-hydroxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-2-methyl)pentyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-ethyl-2-hydroxy)pentyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt; (±)-(2R,3R,5R, 1'R,2'R)-2-( 1 -Acetamido-2,3-dihydroxy)propyl-3-(cis-propen-1 -ylI) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ;. (-)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-ethyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylate Ammonium Salt; (±)-(2R,3S,5R, 1'R,2'R)-2-(1-Acetamido-2,3-dimethoxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(l1-Acetamido-2-methoxy-2-vinyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R, 3S,5R, 1 'S)-2-(1-Acetamido-2-ethyl)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R, 1 'S,3'S)-2-(1 -Acetamido-2-(N-isopropyl-N-methylamino-N oxide))ethyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1 'S,3'S)-2-(1 -Acetamido-2-(N-ethyl-N-methylamino-N-oxide))ethyl 3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid; -33- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-methoxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-methoxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3R,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy)butyl-3-(pyrazol-3-yl) pyrrolidine-5-carboxylic Acid; -34- WO 99/54299 PCT/US99/07945 (±)-(2R,3S, 5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'R)-2-(1-Acetamido-1-(3,6-dihydro-2-H-pyran-2-yl))propyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-methoxy-2-allyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S,3'S)-2-(1 -Acetamido-2-hydroxy-3-methyl)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-methoxy-4-vinyl)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ;. (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy-3-cyano)propyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-1 -(3,6-dihydro-2-H-pyran-2-yl))methyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2,3-dimethoxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S ,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxymethyl-2-hydroxy)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R,1 'R,2'S)-2-(1 -Acetamido-2-ethoxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-dimethyl)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid ; -35- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-ethoxy-3-vinyl)propyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S, 5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-2-(propeny-2-yl))ethyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy)hexyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R, 3S,5R, 1 'S)-2-(1-acetamido-3-methyl)butyl-3-(cis-propen-1-yl)-pyrrolidine 5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy)butyl-3-vinyl-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)pentyl-3-vinyl-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'R)-2-(1-Acetamido-2-hydroxyethyl-2-hydroxy)pentyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'R)-2-(1-Acetamido-2-hydroxy)butyl-3-vinyl-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'R)-2-(1-Acetamido-2-methoxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'R)-2-(1 -Acetamido-2-hydroxy)pentyl-3-vinyl-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R, 1'R)-2-(1-Acetamido-2-hydroxy)ethyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid ; -36- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-(cis-2-chloro-vin-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3R,5R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-(pyrazol-3-yl)-pyrrolidine-5 carboxylic Acid; (±)-(2R,3S,5R, l'S,3'R)-2-(1-Acetamido-3-hydroxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid; (±)-(2R,3R,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(thiazol-4-yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3R,5R,1'S)-1- t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-(thiazol 2-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-vinyl-pyrrolid ine-5-carboxylic Acid ; (±)-(2R,3S,5R, l'S)-2-(1-acetamido-3-methyl)butyl-3-(2,2-difluoro-vin-1 -yl) pyrrolidine-5-carboxylic Acid; (±)-(2R,3R, 5 R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-(pyrazol-3-yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3R,5R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-(isoxazol-3-yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3R,5R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-(isoxazol-5-yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3R,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(imidazol-2-yl)-pyrrolidine-5 Carboxylic Acid; -37- WO 99/54299 PCT/US99/07945 (±)-(2R,3R,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(imidazol-4-yl)-pyrrolidine-5 carboxylic Acid ;and (±)-(2S,3R,5R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-amino-pyrrolidine-5 carboxylic Acid; or a pharmaceutically acceptable salt, ester or prodrug thereof. More preferred compounds of the invention include compounds selected from the group consisting of: (±)-(2R, 3S ,5R, 1' R)-2-(1-Acetamido-2-ethyl-2-hydroxy)butyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-2-methyl)pentyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R, 3S, 5R, 1' R,2'S)-2-(1-Acetamido-2-ethyl-2-hydroxy)pentyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt; (±)-(2R,3R,5R, 1'R,2'R)-2-(1-Acetamido-2,3-dihydroxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ;. (-)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-ethyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylate Ammonium Salt; (±)-(2R,3S,5R, 1'R,2'R)-2-(1-Acetamido-2,3-dimethoxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-methoxy-2-vinyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; -38- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-ethyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R, 1 'S,3'S)-2-(1 -Acetamido-2-(N-isopropyl-N-methylamino-N oxide))ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1 'S,3'S)-2-(1 -Acetamido-2-(N-ethyl-N-methylamino-N-oxide))ethyl 3-(cis-propen- 1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-methoxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-methoxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3R,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy)butyl-3-(pyrazol-3-yl) pyrrolidine-5-carboxylic Acid; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'R)-2-(1 -Acetamido-1 -(3,6-dihydro-2-H-pyran-2-yl))propyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-methoxy-2-allyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S,3'S)-2-(1 -Acetamido-2-hydroxy-3-methyl)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-methoxy-4-vinyl)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-cyano)propyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid ; -39- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-1 -(3,6-dihydro-2-H-pyran-2-yl))methyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2,3-dimethoxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxymethyl-2-hydroxy)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-ethoxy)pentyl-3-(cis-propen- 1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-dimethyl)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S, 5 R, 1'R,2'S)-2-(1 -Acetamido-2-ethoxy-3-vinyl)propyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-2-(propeny-2-yl))ethyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid ; and (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy)hexyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; or a pharmaceutically acceptable salt, ester or prodrug thereof. The term "acid protecting group" as used herein refers to groups used to protect acid groups (for example, -CO 2 H, -SO 3 H, -SO 2 H, -PO 3
H
2 , -PO 2 H groups and the like) against undesirable reactions during synthetic procedures. Commonly used acid protecting groups are disclosed in T.H. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 2nd edition, John Wiley & Sons, New York (1991). Most frequently, such acid protecting groups are esters. -40- WO 99/54299 PCT/US99/07945 Such esters include: alkyl esters, especially loweralkyl esters, including, but not limited to, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, t-butyl, n-pentyl esters and the like; arylalkyl esters including, but not limited to, benzyl, phenethyl, 3 phenylpropyl, naphthylmethyl esters and the like, wherein the aryl part of the arylalkyl group is unsubstituted or substituted as previously defined herein; silylesters, especially, (tri-loweralkyl)silyl esters, (di-loweralkyl)(aryl)silyl esters and (loweralkyl)(di-aryl)silyl esters, including, but not limited to, trimethylsilyl, triethylsilyl, isopropyldimethylsilyl, t-butyldimethylsilyl, methyldiisopropylsilyl, methyldi-t-butylsilyl, triisopropylsilyl, methyldiphenylsilyl, isopropyldiphenylsilyl, butyldiphenylsilyl, phenyldiisopropylsilyl esters and the like; and the like. Preferred acid protecting groups are loweralkyl esters. The term "activated carboxylic acid group" as used herein refers to acid halides such as acid chlorides and also refers to activated ester derivatives including, but not limited to, formic and acetic acid derived anhydrides, anhydrides derived from alkoxycarbonyl halides such as isobutyloxycarbonylchloride and the like, anhydrides derived from reaction of the carboxylic acid with N,N'-carbonyldiimidazole and the like, N-hydroxysuccinimide derived esters, N-hydroxyphthalimide derived esters, N-hydroxybenzotriazole derived esters, N-hydroxy-5-norbornene-2,3-dicarboximide derived esters, 2,4,5 trichlorophenol derived esters, p-nitrophenol derived esters, phenol derived esters, pentachlorophenol derived esters, 8-hydroxyquinoline derived esters and the like. -41- WO 99/54299 PCT/US99/07945 The term "acyl" as used herein, refers to groups having the formula
-C(=O)-R
95 wherein R 95 is hydrogen or an alkyl group. Preferred alkyl groups as
R
9 " are loweralkyl groups. Representative examples of acyl groups include groups such as, for example, formyl, acetyl, propionyl, and the like. The term "acylamino" as used herein, refers to groups having the formula
-NHR
89 wherein R 89 is an acyl group. Representative examples of acylamino include acetylamino, propionylamino, and the like. The term "alkenyl" as used herein, refers to a straight or branched chain hydrocarbon radical containing from 2 to 15 carbon atoms and also containing at least one carbon-carbon double bond. The term "lower alkenyl" refers to straight or branched chain alkenyl radicals containing from 2 to 6 carbon atoms. Representative examples of alkenyl groups include groups such as, for example, vinyl, 2-propenyl, 2-methyl-l-propenyl, 3-butenyl, 4-pentenyl, 5-hexenyl and the like. The term "alkenylene" as used herein, refers to a divalent group derived from a straight or branched chain hydrocarbon containing from 2 to 15 carbon atoms and also containing at least one carbon-carbon double bond. The term "lower alkenylene" refers to a divalent group derived from a straight or branched chain alkene group having from 2 to 6 carbon atoms. Representative examples of alkenylene groups include groups such as, for example, -CH=CH-,
-CH
2 CH=CH-, -C(CH 3 )=CH-, -CH 2
CH=CHCH
2 -, and the like. The term "alkenyloxy" as used herein, refers to groups having the formula
-OR
81 where R 81 is an alkenyl group. The term "alkoxy" as used herein, refers to groups having the formula
-OR
99 wherein R 99 is an alkyl group. Preferred R 99 groups are loweralkyl groups. -42- WO 99/54299 PCT/US99/07945 Representative examples of alkoxy groups include groups such as, for example, methoxy, ethoxy, tert-butoxy, and the like. The term "alkoxyalkoxy" as used herein, refers to groups having the formula -O-R 96
-O-R
97 wherein R 97 is loweralkyl, as defined herein, and R 96 is a lower alkylene group. Representative examples of alkoxyalkoxy groups include groups such as, for example, methoxymethoxy, ethoxymethoxy, t-butoxymethoxy and the like. The term "alkoxyalkyl" as used herein refers to an alkyl radical to which is appended an alkoxy group, for example, methoxymethyl, methoxylpropyl and the like. The term "alkoxycarbonyl" as used herein, refers to groups having the formula, -C(=O)- R 80 , where R 80 is an alkoxy group. The term "alkoxycarbonylalkyl" as used herein, refers to groups having the formula, -C(=O)- R 79 , appended to the parent molecular moiety through an alkylene linkage, where R 79 is an alkoxy group. As used herein, the term "alkyl" refers to straight or branched chain hydrocarbon radicals containing from 1 to 12 carbon atoms. The term "loweralkyl" refers to straight or branched chain alkyl radicals containing from 1 to 6 carbon atoms. Representative examples of alkyl groups include groups such as, for example, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, t-butyl n-pentyl, 1-methylbutyl, 2,2-dimethylbutyl, 2-methylpentyl, 2,2-dimethyl propyl, n-hexyl, and the like. The hydrocarbon chains in alkyl groups or the alkyl portion of an alkyl-containing substituent can be optionally interrupted by one or two heteroatoms or heterogroups independently selected from the group consisting of oxygen, -N(R 27 )- and sulfur wherein R 27 at each occurrence is independently hydrogen, loweralkyl, cylcoalkyl, cycloalkylalkyl or arylalkyl and -43- WO 99/54299 PCT/US99/07945 wherein two such heteroatoms or heterogroups are separated by at least one carbon atom. The term "alkylamino" as used herein, refers to groups having the formula
-NHR
91 wherein R 91 is an alkyl group. Preferred R 91 groups are loweralkyl groups. Representative examples of alkylamino include methylamino, ethylamino, and the like. The term "alkylene" as used herein, refers to a divalent group derived from a straight or branched chain saturated hydrocarbon group having from 1 to 15 carbon. The term "lower alkylene" refers to a divalent group derived from a straight or branched chain saturated hydrocarbon group having from 1 to 6 carbon atoms. Representative examples of alkylene groups include groups such as, for example, methylene (-CH 2 -), 1,2-ethylene (-CH 2
CH
2 -), 1,1-ethylene
(-CH(CH
3 )-), 1,3-propylene (-CH 2
CH
2
CH
2 -), 2,2-dimethylpropylene
(-CH
2
C(CH
3
)
2
CH
2 -), and the like. The hydrocarbon chains in alkylene groups or the alkylene portion of an alkylene-containing substituent can be optionally interrupted by one or two heteroatoms or heterogroups independently selected from the group consisting of oxygen, -N(R 27 )- and sulfur wherein R 2 7 at each occurrence is independently hydrogen, loweralkyl, cylcoalkyl, cycloalkylalkyl or arylalkyl and wherein two such heteroatoms or heterogroups are separated by at least one carbon atom. The term "alkylsulfonyl" as used herein refers to the group having the formula, -SO 2
-R
7 8 , where R 7 8 is an alkyl group. Preferred groups R 78 are loweralkyl groups. The term "alkylsulfonylamino" as used herein refers to the group having the formula, -SO 2
-R
77 , appended to the parent molecular moiety through an amino linkage (-NH-), where R 77 is an alkyl group. Preferred groups R 77 are loweralkyl groups. -44- WO 99/54299 PCT/US99/07945 The term "alkynyl" as used herein, refers to a straight or branched chain hydrocarbon radical containing from 2 to 15 carbon atoms and also containing at least one carbon-carbon triple bond. The term "lower alkynyl" refers to straight or branched chain alkynyl radicals containing from 2 to 6 carbon atoms. Representative examples of alkynyl groups include groups such as, for example, acetylenyl, 1-propynyl, 2- propynyl, 3-butynyl, 2-pentynyl, 1-butynyl and the like. The term "alkynylene" as used herein, refers to a divalent group derived from a straight or branched chain hydrocarbon containing from 2 to 15 carbon atoms and also containing at least one carbon-carbon triple bond. The term "lower alkynylene" refers to a divalent group derived from a straight or branched chain alkynylene group from 2 to 6 carbon atoms. Representative examples of alkynylene groups include groups such as, for example, -C-C-, -CH 2 -C-=C-,
-C-C-CH
2 -, -CH(CH 3 )-C=C-, and the like. The term "aminoalkyl" as used herein refers to an alkyl radical to which is appended an amino (-NH 2 ) group. The term "aryl" as used herein refers to a carbocyclic ring system having 6-10 ring atoms and one or two aromatic rings. Representative examples of aryl groups include groups such as, for example, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl and the like. The aryl groups can be unsubstituted or substituted with one, two or three substituents, each independently selected from loweralkyl, halo, haloalkyl, haloalkoxy, hydroxy, oxo (=O), hydroxyalkyl, alkenyloxy, alkoxy, alkoxyalkoxy, alkoxycarbonyl, alkoxycarbonylalkyl, thioalkoxy, amino, alkylamino, alkylsulfonyl, dialkylamino, acylamino, unsubstituted aryl, unsubstituted arylalkyl, unsubstituted arylalkoxy, unsubstituted aryloxy, mercapto, cyano, nitro, carboxy, carboxaldehyde, NH 2 C(=O)-, cycloalkyl, carboxyalkyl, alkylsulfonylamino, unsubstituted heterocyclic, unsubstituted (heterocyclic)alkyl, unsubstituted -45- WO 99/54299 PCT/US99/07945 (heterocyclic)alkoxy, unsubstituted (heterocyclic)oxy and -SO 3 H. Preferred aryl substituents are each independently selected from the group consisting of loweralkyl, halo, haloalkyl, hydroxy, hydroxyalkyl, alkenyloxy, alkoxy, alkoxyalkoxy, thioalkoxy, amino, alkylamino, dialkylamino, alkylsulfonyl, acylamino, cyano and nitro. Examples of substituted aryl include 3-chlorophenyl, 3-fluorophenyl, 4-chlorophenyl, 4-fluorophenyl, 3,4-dichlorophenyl, 3-chloro-4-fluoro-phenyl, 4-methylsulfonylphenyl, and the like. The term "(aryl)alkenyl" refers to a lower alkenyl group having appended thereto an aryl group. Representative examples of (aryl)alkenyl groups include groups such as, for example phenylethylenyl, phenylpropenyl, and the like. The term "(aryl)alkyl" refers to a loweralkyl group having appended thereto an aryl group. Representative examples of (aryl)alkyl groups include groups such as, for example benzyl and phenylethyl. The term "arylalkoxy" as used herein refers to the group having the formula, -O-R 76 where R 76 is an arylalkyl group. The term "(aryl)alkynyl" refers to an alkynylene group having appended thereto an aryl group. Representative examples of (aryl)alkynyl groups include groups such as, for example phenylacetylenyl, phenylpropynyl, and the like. The term "aryloxy" as used herein refers to the group having the formula,
-O-R
72 , where R 7 2 is an aryl group. The term "carbamoyl" as used herein refers to the group having the formula, -C(=O)-NH 2 . The term "carboxyalkyl" as used herein, refers to the group having the formula, -R4-COOH, where R 64 is a lower alkylene group. The term "cyanoalkyl" as used herein refers to an alkyl radical to which is appended a cyano group (-CN). -46- WO 99/54299 PCT/US99/07945 The term "cycloalkenyl" as used herein refers to an aliphatic ring system having 5 to 10 carbon atoms and 1 or 2 rings containing at least one double bond in the ring structure. Representative examples of cycloalkenyl groups include groups such as, for example, cyclohexene, cyclopentene, norbornene and the like. Cycloalkenyl groups can be unsubstituted or substituted with one, two or three substituents independently selected hydroxy, halo, amino, alkylamino, dialkylamino, alkoxy, alkoxyalkoxy, thioalkoxy, haloalkyl, mercapto, loweralkenyl and loweralkyl. Preferred substitutents are independently selected from loweralkyl, loweralkenyl, haloalkyl, halo, hydroxy and alkoxy. The term "(cycloalkenyl)alkenyl" as used herein refers to a cycloalkenyl group appended to a lower alkenyl radical. Representative examples of (cycloalkenyl)alkenyl groups include groups such as, for example, cyclohexenylethylene, cyclopentenylethylene, and the like. The term "(cycloalkenyl)alkyl" as used herein refers to a cycloalkenyl group appended to a lower alkyl radical. Representative examples of (cycloalkenyl)alkyl groups include groups such as, for example, cyclohexenylmethyl, cyclopentenylmethyl, cyclohexenylethyl, cyclopentenylethyl, and the like. The term "(cycloalkenyl)alkynyl" as used herein refers to a cycloalkenyl group appended to a lower alkynyl radical. Representative examples of (cycloalkenyl)alkynyl groups include groups such as, for example, cyclohexenylacetylenyl, cyclopentenylpropynyl, and the like. The term "cycloalkyl" as used herein refers to an aliphatic ring system having 3 to 10 carbon atoms and 1 or 2 rings. Representative cylcoalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornane, bicyclo[2.2.2]octane and the like. -47- WO 99/54299 PCT/US99/07945 Cycloalkyl groups can be unsubstituted or substituted with one, two or three substituents independently selected hydroxy, halo, amino, alkylamino, dialkylamino, alkoxy, alkoxyalkoxy, thioalkoxy, haloalkyl, mercapto, loweralkenyl and loweralkyl. Preferred substitutents are independently selected from loweralkyl, loweralkenyl, haloalkyl, halo, hydroxy and alkoxy. The term "(cycloalkyl)alkyl" as used herein refers to a cycloalkyl group appended to a loweralkyl radical. Representative examples of (cycloalkyl)alkyl groups include groups such as, for example, cyclohexylmethyl, cyclopentylmethyl, cyclohexylethyl, cyclopentylethyl, and the like. The term "(cycloalkyl)alkenyl" as used herein refers to a cycloalkyl group appended to a lower alkenyl radical. Representative examples of (cycloalkyl) alkenyl groups include groups such as, for example, cyclohexylethylene, cyclopentylethylene, and the like. The term "(cycloalkyl)alkynyl" as used herein refers to a cycloalkyl group appended to a lower alkynyl radical. Representative examples of (cycloalkyl) alkynyl groups include groups such as, for example, cyclohexylacetylenyl, cyclopentylpropynyl, and the like. The term "dialkylamino" as used herein, refers to groups having the formula -N(R 90
)
2 wherein each R 90 is independently a lower alkyl group. Representative examples of dialkylamino include dimethylamino, diethylamino, N methyl-N-isopropylamino and the like. The term "halo" as used herein refers to F, CI, Br or I. The term "haloalkenyl" as used herein refers to a loweralkenyl group in which one or more hydrogen atoms is replaced with a halogen. Examples of haloalkenyl groups include 2-fluoroethylene, 1-chloroethylene, 1,2 difluoroethylene, trifluoroethylene, 1,1,1-trifluoro-2-propylene and the like. -48- WO 99/54299 PCT/US99/07945 The term "haloalkoxy" as used herein refers to the group having the formula, -OR 6 9 , where R 69 is a haloalkyl group as defined herein. Examples of haloalkoxy include chloromethoxy, fluoromethoxy, dichloromethoxy, trifluoromethoxy and the like. The term "haloalkyl" as used herein, refers to a loweralkyl group in which one or more hydrogen atoms has been replaced with a halogen including, but not limited to, trifluoromethyl, trichloromethyl, difluoromethyl, dichloromethyl, fluoromethyl, chloromethyl, chloroethyl, 2,2-dichloroethyl, pentafluoroethyl and the like. The term "heterocyclic ring" or "heterocyclic" or "heterocycle" as used herein, refers to any 3- or 4-membered ring containing a heteroatom selected from oxygen, nitrogen and sulfur; or a 5-, 6- or 7-membered ring containing one, two, three, or four nitrogen atoms; one oxygen atom; one sulfur atom; one nitrogen atom and one sulfur atom; two nitrogen atoms and one sulfur atom; one nitrogen atom and one oxygen atom; two nitrogen atoms and one oxygen atom; two oxygen atoms in non-adjacent positions; one oxygen atom and one sulfur atom in non-adjacent positions; or two sulfur atoms in non-adjacent positions. The 5-membered ring has 0-2 double bonds and the 6- and 7-membered rings have 0-3 double bonds. The nitrogen heteroatoms can be optionally quaternized. The term "heterocyclic" also includes bicyclic groups in which any of the above heterocyclic rings is fused to a benzene ring or a cyclohexane ring or another heterocyclic ring, such as, for example, indolyl, dihydroindolyl, quinolyl, isoquinolyl, tetrahydroquinolyl, tetrahydroisoquinolyl, decahydroquinolyl, decahydroisoquinolyl, benzofuryl, dihydrobenzofuryl or benzothienyl and the like. Heterocyclic groups include, but are not limited to groups such as, for example, aziridinyl, azetidinyl, epoxide, oxetanyl, thietanyl, pyrrolyl, pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, tetrahydropyridyl, piperidinyl, homopiperidinyl, pyrazinyl, -49- WO 99/54299 PCT/US99/07945 piperazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl, oxazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiomorpholinyl, thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, oxetanyl, dihydrofuranyl, tetrahydrofuranyl, dihydropyranyl, tetrahydropyranyl, thienyl, dihydrothienyl, tetrahydrothienyl, triazolyl, triazolinyl, tetrazolyl, tetrazolinyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4 oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, oxadiazolinyl,, 1,2,3 thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, thiadiazolinyl,1,3-dithiolinyl, 1,2-dithiolyl, 1,3-dithiolyl, 1,3-dioxolinyl, didehydrodioxolanyl, 1,3-oxathiolinyl, oxathiolyl, pyrimidyl, benzothienyl and the like. Heterocyclic groups also include compounds of the formula where X* is -OH 2 or -0- and Y* is -C(0)- or [-C(R 92
)
2 -]v where R 92 is hydrogen or C1-C4 alkyl where v is 1, 2, or 3 such as 1,3-benzodioxolyl, 1,4-benzodioxanyl and the like. Heterocyclic groups also include bicyclic rings such as quinuclidinyl and the like. Heterocyclic groups can be unsubstituted or substituted with from one to three substituents, each independently selected from loweralkyl, hydroxy, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino and halogen. In addition, nitrogen containing heterocyclic rings can be N-protected. The term "(heterocyclic)alkenyl" as used herein refers to a heterocyclic group appended to a lower alkenyl radical including, but not limited to, pyrrolidinylethenyl, morpholinylethenyl and the like. -50- WO 99/54299 PCT/US99/07945 The term "(heterocyclic)alkoxy" as used herein refers to the group having the formula, -OR 6 8 , where R 68 is a (heterocyclic)alkyl group. The term "(heterocyclic)alkyl" as used herein refers to a heterocyclic group appended to a loweralkyl radical including, but not limited to, pyrrolidinylmethyl, morpholinylmethyl and the like. The term "(heterocyclic)alkynyl" as used herein refers to a heterocyclic group appended to a lower alkynyl radical including, but not limited to, pyrrolidinylacetylenyl, morpholinylpropynyl and the like. The term "(heterocyclic)oxy" as used herein refers to a heterocyclic group appended to the parent molecular moiety through an oxygen atom (-0-). The term "hydroxy protecting group", "hydroxyl protecting group" or "-OH protecting group" as used herein refers to refers to groups used to hydroxy groups against undesirable reactions during synthetic procedures. Commonly used hydroxy protecting groups are disclosed in T.H. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 2nd edition, John Wiley & Sons, New York (1991). Such hydroxy protecting groups include: methyl ether; substituted methyl ethers, including, but not limited to, methoxymethyl, methylthiomethyl, t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl, benzyloxymethyl, p-methoxybenzyloxymethyl, (4-methoxyphenoxy)methyl, t butoxymethyl, 2-methoxyethoxymethyl, 2,2,2-trichloroethoxymethyl, 2 (trimethylsilyl)ethoxymethyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydrofuranyl, tetrahydrothiofuranyl ether and the like; substituted ethyl ethers, including, but not limited to, 1-ethoxyethyl, 1 methyl-l-methoxyethyl, 1-methyl-l-benzyloxyethyl, 2,2,2-trichloroethyl, trimethylsilylethyl, t-butyl ether and the like; -51- WO 99/54299 PCT/US99/07945 benzyl ether; substituted benzyl ethers, including, but not limited to, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitorbenzyl, p-halobenzyl, p-cyanobenzyl, diphenylmethyl, triphenylmethyl ether and the like; silyl ethers, including, but not limited to, trimethylsilyl, triethylsilyl, triisopropylsilyl, dimethylisopropylsilyl, diethylisopropylsilyl, dimethylthexylsilyl, t butyldimethylsilyl, t-butyldiphenylsilyl, tribenzylsilyl, triphenylsilyl, diphenylmethylsilyl ether and the like; esters, including, but not limited to, formate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, phenoxyacetate, pivaloate, benzoate ester and the like; and the like. Preferred hydroxy protecting groups include substituted methyl ethers, benzyl ether, substituted benzyl ethers, silyl ethers and esters. The term "hydroxyalkyl" as used herein refers to the group having the formula, -R 6 5 -OH, where R 65 is an alkylene group The term "leaving group" as used herein refers to a group which is easily displaced from the compound by a nucleophile. Examples of leaving groups include a halide (for example, Cl, Br or I) or a sulfonate (for example, mesylate, tosylate, triflate and the like) and the like. The term "N-protecting group" or "N-protected" as used herein refers to those groups intended to protect the N-terminus of an amino acid or peptide or to protect an amino group against undesirable reactions during synthetic procedures. Commonly used N-protecting groups are disclosed in T.H. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 2nd edition, John Wiley & Sons, New York (1991). N-protecting groups comprise acyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, -52- WO 99/54299 PCT/US99/07945 trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, a-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and the like; sulfonyl groups such as benzenesulfonyl, p-toluenesulfonyl and the like; carbamate forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-meth oxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxy benzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycar bonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycar bonyl, 1-(p-biphenylyl)-1l-methylethoxycarbonyl, a,a-dimethyl-3,5-dimethoxy benzyloxycarbonyl, benzhydryloxycarbonyl, t-butyloxycarbonyl, diisopropyl methoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyl oxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitro-phen oxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyl oxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl and the like; alkyl groups such as benzyl, triphenylmethyl, benzyloxymethyl and the like; and silyl groups such as trimethylsilyl and the like. Preferred N-protecting groups are formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, phenylsulfonyl, benzyl, t-butyloxycarbonyl (Boc) and benzyloxycarbonyl (Cbz). The term "thioalkoxy" as used herein refers to groups having the formula
-SR
98 wherein R 98 is an alkyl group. Preferred groups R 98 are loweralkyl groups. The term "thio-substituted alkyl" as used herein refers to an alkyl radical to which is appended a thiol group (-SH). As used herein, the terms "S" and "R" configuration are as defined by the IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem. (1976) 45, 13 - 30. The compounds of the invention can comprise asymmetrically substituted carbon atoms. As a result, all stereoisomers of the compounds of the invention -53- WO 99/54299 PCT/US99/07945 are meant to be included in the invention, including racemic mixtures, mixtures of diastereomers, as well as individual optical isomers, including, enantiomers and single diastereomers of the compounds of the invention substantially free from their enantiomers or other diastereomers. By "substantially free" is meant greater than about 80% free of other enantiomers or diastereomers of the compound, more preferably greater than about 90% free of other enantiomers or diastereomers of the compound, even more preferably greater than about 95% free of other enantiomers or diastereomers of the compound, even more highly preferably greater than about 98% free of other enantiomers or diastereomers of the compound and most preferably greater than about 99% free of other enantiomers or diastereomers of the compound. In addition, compounds comprising the possible geometric isomers of carbon-carbon double bonds and carbon-nitrogen double are also meant to be included in this invention. Individual stereoisomers of the compounds of this invention can be prepared by any one of a number of methods which are within the knowledge of one of ordinary skill in the art. These methods include stereospecific synthesis, chromatographic separation of diastereomers, chromatographic resolution of enantiomers, conversion of enantiomers in an enantiomeric mixture to diastereomers and then chromatographically separating the diastereomers and regeneration of the individual enantiomers, enzymatic resolution and the like. Stereospecific synthesis involves the use of appropriate chiral starting materials and synthetic reactions which do not cause racemization or inversion of stereochemistry at the chiral centers. -54- WO 99/54299 PCT/US99/07945 Diastereomeric mixtures of compounds resulting from a synthetic reaction can often be separated by chromatographic techniques which are well-known to those of ordinary skill in the art. Chromatographic resolution of enantiomers can be accomplished on chiral chromatography resins. Chromatography columns containing chiral resins are commercially available. In practice, the racemate is placed in solution and loaded onto the column containing the chiral stationary phase. The enantiomers are then separated by HPLC. Resolution of enantiomers can also be accomplished by converting the enantiomers in the mixture to diastereomers by reaction with chiral auxiliaries. The resulting diastereomers can then be separated by column chromatography. This technique is especially useful when the compounds to be separated contain a carboxyl, amino or hydroxyl group that will form a salt or covalent bond with the chiral auxiliary. Chirally pure amino acids, organic carboxylic acids or organosulfonic acids are especially useful as chiral auxiliaries. Once the diastereomers have been separated by chromatography, the individual enantiomers can be regenerated. Frequently, the chiral auxiliary can be recovered and used again. Enzymes, such as esterases, phosphatases and lipases, can be useful for resolution of derivatives of the enantiomers in an enantiomeric mixture. For example, an ester derivative of a carboxyl group in the compounds to be separated can be prepared. Certain enzymes will selectively hydrolyze only one of the enantiomers in the mixture. Then the resulting enantiomerically pure acid can be separated from the unhydrolyzed ester. In addition, solvates and hydrates of the compounds of Formula I and II and Ill are meant to be included in this invention. -55- WO 99/54299 PCT/US99/07945 When any variable (for example R 1 , R 2 , R 3 , m, n, etc.) occurs more than one time in any substituent or in the compound of Formula I or II or III or any other formula herein, its definition on each occurrence is independent of its definition at every other occurrence. In addition, combinations of substituents are permissible only if such combinations result in stable compounds. Stable compounds are compounds which can be isolated in a useful degree of purity from a reaction mixture. This invention is intended to encompass compounds having Formula I and II and III when prepared by synthetic processes or by metabolic processes. Preparation of the compounds of the invention by metabolic processes include those occurring in the human or animal body (in vivo) or processes occurring in vitro. Compounds of the invention can be prepared according to the methods described in Schemes 1-5 as shown below. Throughout the schemes, methods will be illustrated wherein R 1 is a carboxylic acid or carboxylic acid ester substituent. It will be understood by those skilled in the art that other R 1 substituents can (a) be obtained either from the carboxylic acid or carboxylic acid ester group, (b) can be introduced by similar methods to those used to introduce the carboxylic acid or carboxylic acid ester group or (c) can be introduced by other methods generally known in the art. In adddition, throughout the schemes, methods will be illustrated wherein
R
4 , R 6 , R 7 , R 8 , R 9 and R 1 0 are hydrogen. It will be understood by those skilled in the art that compounds wherein one or more of these substituents is other than hydrogen can be prepared by methods analogous to those disclosed in the schemes or by other methods generally known in the art. -56- WO 99/54299 PCT/US99/07945 In addition, throughout the schemes, methods will be illustrated for obtaining compounds of the invention having the preferred relative stereochemistry. It will be understood by those skilled in the art that compounds of the invention having other relative stereochemistry can be prepared by methods analogous to those disclosed in the schemes or by other methods generally known in the art. In addition, throughout the schemes, methods will be illustrated wherein X is -C(=O)-NH-. It will be understood by those skilled in the art that other X groups can be prepared by methods analogous to those disclosed in the schemes or by other methods generally known in the art. As shown in Scheme 1, reaction of acrolein with an N-protected a-amino acid ester 1 (P 1 is an N-protecting group, preferably a benzyl group or the like and P 2 is a carboxylic acid protecting group, preferably a t-butyl group or the like) in an inert solvent (for example, toluene and the like) in the presence of an acid catalyst (for example, acetic acid and the like), followed by equilibration with a base (for example, with triethylamine or the like) and separation of the isomers by chromatography, provides substituted pyrrolidine 2. Reduction of the aldehyde group to an alcohol with an aldehyde to alcohol reducing agent (for example, sodium borohydride or the like) in an inert solvent (for example, methanol or the like), followed by chromatographic separation of the isomers provides alcohol 3. Alcohol 3 can be protected with an hydroxy protecting group P 3 (preferably with a silyl protecting group, for example, t-butyldimethylsilyl or the like) using standard alcohol protection methods to provide 4. Oxidation of the vinyl group of compound 4 to an aldehyde is accomplished by reacting compound 4 with OsO 4 and N-methylmorpholine N-oxide to give the corresponding diol. The diol is then treated with sodium periodate to provide aldehyde 5. Substituents R 3 can be introduced via reaction of aldehyde 5 with a Grignard reagent (for example,
R
3 MgBr or the like) to give alcohol 6. Oxidation of alcohol 6 (for example, Swern -57- WO 99/54299 PCT/US99/07945 oxidation or the like) provides ketone 7. Reductive amination of ketone 7 (for example, by reaction with ammonium acetate and sodium cyanoborohydride in methanol or the like) gives amine 8. Amine 8 can be further functionalized to complete the introduction of the R 2 -X- substituent (for example, by reaction of the amine with an acylating agent such as acetic anhydride or the like or by other acylation methods), followed by chromatographic separation of the diastereomers to give 9a. The other diastereomeric amine (9b) can also be isolated and further transformed according to Scheme 1. Removal of hydroxy protecting group P 3 (for example, by reaction with a fluoride ion source, such as tetrabutylammonium fluoride or the like, when P 3 is a silyl protecting group) provides alcohol 10. Transformation of the hydroxy group of alcohol 10 allows introduction of various substituents Y. For example, alkylation of the hydroxy group provides ethers 11. N deprotection (for example, where P 1 is a benzyl group, by hydrogenation) gives 12', followed by ester hydrolysis (for example, with acid such as HCI), provides compound 12" of the invention. Oxidation of the hydroxy group of 10 (for example, Swern oxidation or the like) provides aldehyde 13. Oxidation of aldehyde 13 (for example, with NaCIO 2 or the like) provides carboxylic acid 14. The carboxylic acid substituent of 14 can be used to introduce a variety of other functional groups in substituent Y. For example, the carboxylic acid can be esterified (for example, by reaction with diazomethane or with ethanol and DCC or the like) or the carboxylic acid or an activated derivative thereof can be reacted with amines to provide 15 (wherein
-C(=O)-R
2 2 represents an ester or an amide). N-deprotection (for example, where P 1 is a benzyl group, by hydrogenation) gives 16', followed by ester hydrolysis (for example, with acid such as HCI), provides compound 16" of the invention. -58- WO 99/54299 PCT/US99/07945 Derivatives of the aldehyde group of 13 or the carboxylic acid group of 14 can be used to introduce substituents Y which are -CN or various heterocycles, according to methods known to those skilled in the art and according to the specific methods exemplified herein. Reaction of aldehyde 13 with loweralkyl- or loweralkenyl-Grignard reagents, followed by oxidation (for example, Swern oxidation or the like), provides ketones 17 wherein R 22 is loweralkyl or loweralkenyl. N-deprotection (for example, where P 1 is a benzyl group, by hydrogenation) gives 18', followed by ester hydrolysis (for example, with acid such as HCI), provides compound 18" of the invention. Compounds wherein substituent Y is an amino group or a derivative of an amino group can be prepared as shown in Scheme 2. Oxidation of aldehyde 2 (for example, with AgO or NaCIO 2 or the like) provides carboxylic acid 19. Curtius rearrangement of carboxylic acid 19 (for example, reaction with DPPA, Et 3 N and benzyl alcohol or the like), followed by chromatographic separation of the diastereomers, provides amide 20 wherein P 4 is an N-protecting group (for example, benzyloxycarbonyl or the like). Transformations analogous to those which converted compound 4 to compound 9a and 9b in Scheme 1, enable the conversion of 20 to 21a and 21b, which can be separated by chromatography. Removal of protecting group P 4 (for example, by selective hydrogenation) provides 22. Further derivatization of the amino group allows for introduction of substituents Y which are amine derivatives. N-deprotection (for example, where P' is a benzyl group, by hydrogenation), followed by ester hydrolysis (for example, with acid such as HCI), provides compounds of the invention wherein Y is amino or an amine derivative. Olefination of aldehyde 13 (for example, with Ph 3
PCH
2 or the like), followed by hydrogenation (causing N-deprotection (for example, where P1 is a benzyl group) and olefin saturation, followed by ester hydrolysis (for example, -59- WO 99/54299 PCT/US99/07945 with acid such as HCI), provides compounds of the invention wherein Y is loweralkyl. As shown in Scheme 3, oxidation of the vinyl group of compound 4 to a diol (for example, with OsO4 and N-methylmorpholine N-oxide or the like) gives diol 23. Removal of N-protecting group P1 (for example, where P 1 is a benzyl group, by hydrogenation) provides pyrrolidine 24. Reprotection with an acid labile N-protecting group P 5 (for example, t-butoxycarbonyl or the like) provides 25. Transformation of compound 25 to aldehyde 26a and 26b can be accomplished in a manner analogous to conversion of compound 4a to compound 10 and compound 10 to compound 13 as shown in Scheme 1. 26a and 26b can be separated by chromatography. Olefination of 26a (for example, with Ph 3
PCH
2 , or triphenylphosine/methylene chloride/n-BuLi, or I-Ph 3
P+CH
2
CH
3 /KOtBu, or the like) provides 27 wherein Y is an olefinic substituent. N-deprotection of the p 5 protecting group and ester hydrolysis, under acidic conditions, provides compounds of the invention 28 wherein Y is an olefinic substituent. In yet another alternative method shown in Scheme 4, the hydroxy group of alcohol 3 is protected with a base-labile hydroxy protecting group P 6 (for example, acetyl or the like) to give compound 29. Oxidation of the vinyl group of 29 with OsO4 and N-methylmorpholine N-oxide provides diol 30. Removal of the
P
1 protecting group (for example, by hydrogenation or the like) provides pyrrolidine 31. Reprotection with an acid-labile N-protecting group p 5 (for example, t-butoxycarbonyl or the like) provides 32. Selective protection of the primary alcohol of 32 with a hydroxy protecting group p 7 (for example, a silyl protecting group such as triisopropylsilyl or the like) provides compound 33. Oxidation of 33 (for example, Swern oxidation or the like) provides ketone 34. Reductive amination of ketone 34 (for example, by reaction with ammonium acetate and sodium cyanoborohydride in methanol or the like) gives amine 35. -60- WO 99/54299 PCT/US99/07945 Amine 35 can be further functionalized to complete the introduction of the R 2
-X
substituent (for example, by reaction of the amine with an acylating agent such as acetic anhydride or the like or by other acylation methods), followed by chromatographic separation of the diastereomers to give 36a. The other diastereomeric amine (36b) can also be isolated and further transformed according this scheme. Selective removal of the P 6 hydroxy protecting group in 36a (for example, with K 2 C0 3 in methanol or the like) provides alcohol 37. Oxidation of the alcohol to an aldehyde (for example, Swern oxidation or the like) provides 38. The aldehyde can serve as a precursor for various substituents Y in the compounds of the invention. For example, olefination of 38 (for example, with Ph 3
PCH
2 , or triphenylphosine/methylene chloride/n-BuLi, or I-Ph 3
P'CH
2 CHa/KOtBu, or the like) provides 39 wherein Y is an olefinic substituent. Removal of the p 7 hydroxy protecting group (for example, with a fluoride ion source such as tetrabutylammonium fluoride or the like) gives alcohol 40. The alcohol can serve as a precursor for a variety of R 3 substituents in the compounds of the invention. For example, the alcohol of 40 can be oxidized to an aldehyde (for example, by Dess-Martin oxidation or the like) to give 41. Aldehyde 41 can be reacted with Grignard reagents (R 14 MgBr or the like) or other organometallic reagents (for example, organolithium reagents such as R 14 Li or the like) to provide 42 as a mixture of alcohol diastereomers which can be separated chromatographically to provide the major isomer 42a and the other isomer 42b. Isomer 42a or the mixture of isomers 42 can be oxidized (for example, by Dess-Martin oxidation or the like) to give ketone 43. Reduction of ketone 43 (for example, with sodium borohydride in ethanol or the like) provides alcohol 42b as the major isomer, which can be isolated by chromatography. N deprotection of the p 5 protecting group and ester hydrolysis, under acidic -61- WO 99/54299 PCT/US99/07945 conditions, provides compounds of the invention 44a or 44b, respectively, wherein Y is an olefinic substituent. Alkylation of alcohol 42a or 42b provides ethers 45a or 45b, respectively. N-deprotection of the p 5 protecting group and ester hydrolysis, under acidic conditions, provides compounds of the invention 48a or 48b, respectively, wherein Y is an olefinic substituent. As shown in Scheme 5, reaction of ketone 43 with with Grignard reagents
(R
37 aMgBr or the like) or other organometallic reagents (for example, organolithium reagents such as R 37 aLi or the like) provides alcohols 46a and 46b as a mixture of alcohol diastereomers which can be separated chromatographically. N-deprotection of the P 5 protecting group and ester hydrolysis, under acidic conditions, provides compounds of the invention 47a or 47b, respectively, wherein Y is an olefinic substituent. Alkylation of alcohol 46a or 46b provides ethers 49a or 49b, respectively. N-deprotection of the P 5 protecting group and ester hydrolysis, under acidic conditions, provides compounds of the invention 50a or 50b, respectively, wherein Y is an olefinic substituent. Esters or prodrugs of the compounds of the invention can be prepared by methods known in the art. -62- WO 99/54299 PCT/US99/07945 SCHEME I H 0 HO0 0 0 H _p 1 0 3 p 3 0-, p 3 o-, p 3 0-, p2 HO
OP
2 HOP 2 N Th' NJ N; ~" p 1 0 HO P 1 0 0 p, 1 O0 4 4a 5 p3o HO N~OP 0Op H 2 N OP 2 p 1 1 R p 6 7 8 p~o-,p 3 O-,. R 2C(O)HN/, N,,, 11 /op2 R 2 C(O)HN O 8R 3H 11 0 +R 3Hp0 9a 9b -63- WO 99/54299 PCTIUS99/07945 SCHEME 1 (cont'd.) HO-C. 20 9a R 2 C(O)HN, ,,I 1 /Op 2
R
2 C(O)HN,,, ", 1 /Op2
R
3
P
1 R3 al 10 11
R
20 0-- R0-,
R
2 C(O)HN, ."(p 2
R
2 C(O)HN,,,. OH HO 3 HO 12' 12"' O 0 HO-',/ 10 RC(O)N~ ~~~iOP 2
R
2 C(O)HN,, <i-P
R
3 p 1 0 R 3 P 1a 13 14 0 R /,. R22-- 4 i
R
2 C(O)HN, "" .JOi 2
-R
2 C(O)HN,,, 2 14 - -r 3 Ki rJ i R31 0 HO 15 16' R22C(O)- is an ester or amide -64- WO 99/54299 PCT/US99/07945 SCHEME 1 (cont'd.) 0 R22~ 4
R
2 C(O)HN,,, OH 16' N- "[1' 16 -R 3 0 16"' 02 0 R -A/ R224'/ _ C(O)HN,,OP 2 R C(O)HN,,. 17 18'
R
22 iS loweralkyl or loweralkenyl 0 R22 4
R
2 C(O)HN,,, OH 18' - r N "if 18"9 -65- WO 99/54299 PCT/US99/07945 SCHEME 2 0 HO p H N,
S-P
1 00 19 20 P 4 H N, p 4 HN, R 2C(O)HN,,, p R 2C(O)HN p 20 r-N +N R 3 1 R R 3 l 21 a 21 b
H
2 N. H 2 N, 22N R 2C(O)HN,, R 2 C(O)HN oip 2 R 3 01 R 0 22a 22b -66- WO 99/54299 PCT/US99/07945 SCHEME 3
P
3 0-, p 3 0 4 HO . OP 2 HO .,, OP 2 HO 0 O 23 24 p30 HO OP 2 24R 3 N 5 if
P
5 P0 25 0 0 H4 H R C(O)HN, ,, OP 2 R2C(O)HN Op 2 25 .. r- -C )i*') R p 50 P3 5 0 26a 26b Y-1. Y-1: 27 28 Y is an alkene -67- WO 99/54299 PCT/US99/07945 SCHEME 4 p60-, p60 S OP 2 HO -,OP 2 pH0O p 1 0 29 30 p 6 0-, p 6 0 H ', OP HO. ., OP 2 30 HO N OpHO N 0 15 HO H 0 HO p 5 0 31 32 p 6 O-. p 6
O--
HOH Op 2 ., 32 NT,:1 -o. ..f 5~ "<r N 0i p o 5 0 p 7 o p5 33 34 p 6 o-. 34
H
2 N 'If OP2 p 7 0 p 5 0 35 -68- WO 99/54299 PCT/US99/07945 SCHEME 4 (cont'd.) p6o-, P60O.. R2C(O)HN,, OP 2
R
2 C(O)HN Op2 35 N 'f + P "'i 7 5 7 P0 p O p70 p O 36a 36b 0 HO-. H R2C(O)HN,,,. OP 2 R2C(O)HN,,,, P 2 36 N Ni 36a 'p70 p "I
O
r"'""N " o
P
5 ~ 0 P0 ' -70 P5 O 70" O 37 38 R2C(O)HN OP 2
R
2 C(O)HN, OP 2 38 - TIi,. I Sp 7 0 p 5 O HO P 5 O 39 Y is an alkene 40 or haloalkene Y. 40 R 2C(O)HN ' , ,Op 2 41 -69- WO 99/54299 PCT/US99/07945 SCHEME 4 (cont'd.) R2C(O)HN, OP2 R 2 C(O)HN,,, Op 2 41N N'i 14 -,, OH 0 + 14 OH P Q0 R /OH R OH 42a 42b II
R
2 C(O)HN,, Op 2
R
2 C(O)HN,, Op 2 N N
R
14
-P
5 P14 5 bR37a
OR
3 7 a 45a 45b ITI
R
2 C(O)HN,, OH R 2 C(O)HN,,. . OH N N
R
14 HO 14 H O
OR
37a
OR
37 a 48a 48b -70- WO 99/54299 PCT/US99/07945 SCHEME 4 (cont'd.) Y-1 Y 42a R 2 C(O)HN-, N- OP 2
R
2 C(O)HN, OP2 42a C 14 05 0 14 p 5 O R o R OH 43 42b R OH H 0 R 2 C(O)HN, OH 44a R14OHH 0 44b -71- WO 99/54299 PCT/US99/07945 SCHEME 5
R
2 C(0)HN,. OP 2
R
2 C(0)HN ,,OP2 N NY R14 p O + 14 . O R OH R R37a
R
37 a bH 46a 46b 2 2 Y-1. R C(O)HN, OH R2C(O)H N,D OH R14 OHHO R 14 H O
R
37 a
R
37 a OH 47a 47b -72- WO 99/54299 PCT[US99/07945 SCHEME 5 (cont'd.) Y, ~Y R 2
C(O)HN
4 ,1' N p R.,O R 2 C(O)HN,,. OHR 14 p +
R
14 5 R 37 a R 37 a -0OH 46a 46 b II
R
2
C(O)HN,
4 op 2
R
2 C(O)HN,, 14 0 o 1
R
3 7 a OR 37 C
R
7 a r 37 c 49a 49 b I I R 2 C(O)H N,,. OH R 2 C(O)H N,,, OH 14 H0 14 HO0 r ., R 37c 37c
R
3 7 a R 3cR 37 a ORc 50a 50b -73- WO 99/54299 PCTIUS99/07945 The other compounds of the invention can be readily prepared from the compounds described herein using techniques known in the chemical literature. The methods required are known and can be readily practiced by those having ordinary skill in the art. Key intermediates for the preparation of compounds of the invention include the following: (1) o H
OP
2
P
1 0 wherein P 1 is an N-protecting group (preferably, a benzyl group or a substituted benzyl group) and P 2 is a carboxylic acid protecting group (preferrably, a loweralkyl group, especially t-butyl); preferrably, P 1 and P 2 can be selectively deprotected/removed; or a salt thereof; (2) p3,"OP P , O wherein P 1 is an N-protecting group (preferably, a benzyl group or a substituted benzyl group) and P 2 is a carboxylic acid protecting group (preferably, a -74- WO 99/54299 PCT/US99/07945 loweralkyl group, especially t-butyl); and P 3 is hydrogen or a hydroxy protecting group (preferably, an acyl protecting group, for example, acetyl and the like, or a silyl protecting group, for example, t-butyldimethylsilyl and the like); preferrably, p 1 , p 2 and P 3 can be selectively deprotected/removed; or a salt thereof; (3) p 4 HN. Op 2 IO N 0 wherein P 1 is an N-protecting group (preferably, a benzyl group or a substituted benzyl group) and P 2 is a carboxylic acid protecting group (preferably, a loweralkyl group, especially t-butyl); and P 4 is hydrogen or an N-protecting group (preferably, a carbamate N-protecting group, for example, benzyloxycarbonyl and the like); preferrably, P 1 , P 2 and P 4 can be selectively deprotected/removed; or a salt thereof; (4) p60- " PO--, HO OP2 P7a
P
5 0 -75- WO 99/54299 PCT/US99/07945 P6O-,, 0 OP 2 71 p 5 0 or p6O-,
H
2 N
OP
2 PO5 p 7 0 P 5 0 wherein p 5 is an N-protecting group (preferably, an acid labile N-protecting group, such as t-butyloxycarbonyl and the like) and P 2 is a carboxylic acid protecting group (preferably, a loweralkyl group, especially t-butyl); and p 6 is hydrogen or a hydroxy protecting group (preferably, a base labile hydroxy protecting group, such as acetyl and the like); and p 7 is hydroxy protecting group (preferably, a silyl protecting group, such as triisopropylsilyl and the like); preferrably, P 2 , p 5 , p 6 and P 7 can be selectively deprotected/removed; or a salt thereof; and -76- WO 99/54299 PCT/US99/07945 (5) p60-, R2C(O)HN,,, OP 2 N 1~5 1 P70 r ps O or O 0 2 R2C(O)HN,, , Op 2 p 7 0 p 5 0 wherein p 5 is an N-protecting group (preferably, an acid labile N-protecting group, such as t-butyloxycarbonyl and the like) and P 2 is a carboxylic acid protecting group (preferably, a loweralkyl group, especially t-butyl); and P 6 is hydrogen or a hydroxy protecting group (preferably, a base labile hydroxy protecting group, such as acetyl and the like); and p 7 is hydroxy protecting group (preferably, a silyl protecting group, such as triisopropylsilyl and the like); and R 2 is defined as herein (preferably, loweralkyl or haloloweralkyl; most preferably, methyl or trifluoromethyl); preferrably, P 2 , p 5 , p 6 and p 7 can be selectively deprotected/removed; or a salt thereof. -77- WO 99/54299 PCT/US99/07945 All patents, patent applications, and literature references cited in this specification are hereby incorporated by reference in their entirety. In the case of inconsistencies, the present disclosure, including definitions, will prevail. The reagents required for the synthesis of the compounds of the invention are readily available from a number of commercial sources such as Aldrich Chemical Co. (Milwaukee, WI, USA); Sigma Chemical Co. (St. Louis, MO, USA); and Fluka Chemical Corp. (Ronkonkoma, NY, USA); Alfa Aesar (Ward Hill, MA 01835-9953); Eastman Chemical Company (Rochester, New York 14652-3512); Lancaster Synthesis Inc. (Windham, NH 03087-9977); Spectrum Chemical Manufacturing Corp. (Janssen Chemical) (New Brunswick, NJ 08901); Pfaltz and Bauer (Waterbury, CT. 06708). Compounds which are not commercially available can be prepared by employing known methods from the chemical literature. The following examples will serve to further illustrate the preparation of the compounds of the invention, without limitation. -78- WO 99/54299 PCT/US99/07945 Example (±)-(2R,3R, 5R, 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine 5-carboxylic Acid Hydrochloride. O H OtBu N 11 Ko Ph 1 A. (±)-(2S,3R,5R)- and (±)-(2S,3S,5R)-1-Benzyl-2-vinyl-3-formyl-pyrrolidine 5-carboxylic Acid t-Butyl Ester (8:1 ratio). Acrolein (8 mL, 120 mmole) was added to a solution of t-butyl N-benzyl glycinate (4.34 g, 19.6 mmole) and acetic acid (5 drops) in toluene (100 mL). The solution was heated at reflux. After 1 hour, the reaction was cooled to about 50 oC and an additional 3 mL of acrolein were added. The reaction was heated at reflux for an additional 2 hours and concentrated in vacuo. The residue was purified by chromatography on silica gel using 5% ethyl acetate/hexanes to provide a mixture of (±)-(2S,3R,5R)- and (±)-(2S,3S,5R)-1-benzyl-2-vinyl-3 formyl-pyrrolidine-5-carboxylic acid t-butyl esters as an oil (yield: 2.78 g, 45%). The mixture of aldehydes was equilibrated to an 8:1 ratio by stirring the crude product with triethylamine (0.5 mL) in ethyl acetate at room temperature followed by evaporation of the solvents. IH NMR (CDCI3)(major isomer only): 81.45 (s, 9H), 2.26 (m, 1 H), 2.69 (m, 1H), 3.49 (dd, J=7.8, 3.0 Hz, 1H), 3.61 (d, J=13.5 Hz, 1H), 3.93 (m, 1H), 3.94 (d, J=13.5 Hz, 1H), 5.22-5.33 (two dd, 2H), 5.7 (ddd, J=17.7, 10.2, 7.8 Hz, 1H), 7.21-7.35 (m, 5H), 9.71 (d, J=1.2 Hz, 1H). MS (M+H)+ = 316. -79- WO 99/54299 PCT/US99/07945 HO-, ., OtBu OI Ph 1 B. (±)-(2S.3R,5R)-1l-Benzyl-2-vinyl-3-(hydroxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of the 8:1 mixture of (±)-(2S,3R,5R)- and (±)-(2S,3S,5R)-1 benzyl-2-vinyl-3-formylpyrrolidine-5-carboxylic acid t-butyl ester (6.0 g, 19.0 mmole), prepared according to the method described in Example 1A, in 100 mL of methanol was cooled to 0 oC and treated with sodium borohydride (0.72 g, 19.0 mmole). The mixture was stirred for 0.5 hour, warmed to room temperature, and stirred for an additional 1 hour. The reaction was quenched with aqueous ammonium chloride, and the solvent was evaporated. The residue was parti tioned between ethyl acetate and water. The organic layer was dried over MgSO 4 , filtered and concentrated in vacuo. The residual oil was purified by chromatography on silica gel using a gradient of 20-30% ethyl acetate/hexanes to furnish the title compound as a colorless oil (yield: 4.0 g, 66%). 1 H NMR (CDCl3): 61.46 (s, 9H), 1.80 (m, 1H), 2.16 (m, 1H), 2.39 (m, 1H), 2.54 (m, 1H), 3.48-3.53 (m, 2H), 3.08 (d, 2H), 3.91 (d, 2H), 5.17-5.22 (m, 2H), 5.70 (m, 1H), 7.23-7.34 (m, 5H). MS (M+H) + = 318. -80- WO 99/54299 PCT/US99/07945 TBDMSO--, , OtBu Ph 1 C. (±)-(2S,3R,5R)-1l-Benzyl-2-vinyl-3-(t-butyldimethylsilvloxymethyl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2S,3R,5R)-1l-benzyl-2-vinyl-3-(hydroxymethyl) pyrrolidine-5-carboxylic acid t-butyl ester (3.6 g, 11.4 mmole), tert-butyldimethylsilyl chloride (3.7 g, 24.5 mmole) and imidazole (2.8 g, 41.2 mmole) in 80 mL of DMF was stirred at room temperature for 1.5 hours. The reaction was diluted with ethyl acetate, washed with water and brine, dried over MgSO 4 , and concentrated in vacuo. The residue was purified by chromatography on silica gel using 5% ethyl acetate/hexanes to provide the title compound, as a colorless oil (yield: 3.5 g, 71%). 1 H NMR (CDCI3): 80.02 (d, 6H), 0.86 (s, 9H), 1.43 (s, 9H), 1.67(ddd, 1H), 2.11 (m, 1H), 2.28 (m, 1H), 3.40-3.70 (m, 6H), 3.90 (d, 2H), 5.11-5.19 (m, 2H), 5.69 (ddd, 1H), 7.20-7.30 (m, 5H). MS (M+H) = 432. TBDMSO-, H , OtBu 0 Ph 1 D. (±)-(2R, 3R,5R)-1 -Benzyl-2-formyl-3-(t-butyldimethylsilyloxymethyl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. Osmium tetroxide (20 mg) was added to a room temperature solution of (±)-(2S ,3R,5R)-1-benzyl-2-vinyl-3-(t-butydimethylsilyloxymethyl)-pyrrolidine-5 -81- WO 99/54299 PCT/US99/07945 carboxylic acid t-butyl ester (3.5 g, 8.12 mmole) in 60 mL of 8:1 acetone/water and N-methylmorpholine N-oxide (3.0 g, 25.6 mmole). The reaction mixture was stirred at room temperature for 6 hours and quenched with saturated aqueous Na 2
S
2 0 3 . The mixture was stirred for an additional 10 minutes and the solvent removed. The brownish residue was partitioned between dichloromethane and water. The organic layer was dried over MgSO 4 and concentrated in vacuo to provide the intermediate diol as an oil (~3.8g ) which was used without additional purification. MS (crude): (M+H)* = 466 The crude diol was dissolved in 6:1 tetrahydrofuran (THF)/water (50 mL) and treated with sodium periodate (3.0 g, 14.0 mmole). The mixture was stirred at room temperature for 1 hour and diluted with ethyl acetate, washed with water, dried over MgSO 4 , filtered, and concentrated in vacuo. The crude aldehyde was purified by chromatography on silica gel using 3% ethyl acetate/hexanes to provide the title compound as a colorless oil (yield: 1.6 g, 46%). 1 H NMR (CDCI3): 50.03 (d, 6H), 0.86 (s, 9H), 1.46 (s, 9H), 1.72 (m, 1H), 2.26-2.45 (m, 2H), 3.53-3.71 (m, 5H), 3.84 (d, 1H), 3.93 (d, 1H), 7.27-7.31 (m, 5H), 9.32 (d, 1H). MS (M+H)* = 434. -82- WO 99/54299 PCT/US99/07945 TBDMSO-, 0 OtBu 00 Ph 1E. (±)-(2R,3R,5R)-1l-Benzyl-2-(1-oxo-3-ethyl)pentyl-3-(t-butyldimethylsilyloxy methyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A dry flask containing magnesium (0.14 g, 5.83 mmole), under argon, was charged with 10 mL of dry THF and 3 drops of dibromoethane. This was followed by addition of 1-bromo-2-ethylbutane (0.95 g, 5.83 mmole). The reaction mixture was heated at reflux for 45 minutes, until most of the magnesium had reacted. The reaction mixture was cooled to -30 OC and (±)-(2R,3R,5R)-1-benzyl-2-formyl 3-(t-butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (0.5 g, 1.15 mmole) in of THF (6 mL) was added, dropwise. The reaction was slowly warmed to -10 oC, over a period of about 2 hours, and quenched with aqueous ammonium chloride. The resultant slurry was diluted with ethyl acetate and washed with water, brine, and dried over MgSO 4 and concentrated. The crude alcohol product, an oil (-0.85 g), was used without further purification. MS (M+H) + = 520. A solution of oxalyl chloride (2.5 mL, 2M in CH 2
CI
2 ) in 10 mL of anhydrous dichloromethane was prepared and maintained under a nitrogen atmosphere, at -78 *C. DMSO (0.77 mL, 9.83 mmole) was added slowly to the solution. The mixture was stirred for 15 minutes and treated with the crude alcohol prepared above, about 0.85 g, in 5 mL of anhydrous dichloromethane. The solution was stirred for 1 hour and triethylamine (2.3 mL, 16.4 mmole) was added slowly to the reaction mixture. The solution was then allowed to slowly warmed to room temperature and diluted with dichloromethane. The organic layer was washed with water, dried over MgSO 4 , and concentrated. The residue was purified by -83- WO 99/54299 PCT/US99/07945 chromatography on silica gel using 3% ethyl acetate/hexanes to provide the title compound, as an oil (yield: 0.35 g, 66%). MS (M+H)* = 518. TBDMSO-,,.
H
2 N OtBu Ph 1F. (±)-(2R,3R,5R,1'R)- and (±)-(2R,3R,5R,1'S)-1l-Benzyl-2-(1-amino-3-ethyl) pentyl-3-(t-butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R)-1 -benzyl-2-(1-oxo-3-ethyl)pentyl-3-(t-butyl dimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (0.20 g, 0.39 mmole), ammonium acetate (30 equiv.) and sodium cyanoborohydride (10 equiv.) in 5 mL of methanol was heated at reflux for 24 hours with occasional addition of an additional 60 equivalents of ammonium acetate and 20 equivalents of sodium cyanoborohydride. The solvent was evaporated. The resultant residue was partitioned between dichloromethane and water. The organic layer was dried over MgSO 4 , filtered, and concentrated. The product was purified by chromatography on silica gel using 30-50% ethyl acetate/hexanes to provide the title compound as a colorless oil (yield: 130 mg, 64%). 1 H NMR (CDCI 3 ) 8 7.30 (m, 5H), 4.91 (s, 1H), 3.53(m, 2H), 3.08 (m, 1H), 2.88 (m, 1H), 2.35 (m, 1H), 1.85 (m, 1H), 1.44 (s, 9H), 1.20-1.40 (m, 7H), 0.88 (s, 9H), 0.85 (m, 6H), 0.03 (s, 6H) MS (M+H)* = 519. -84- WO 99/54299 PCT/US99/07945 TBDMSO--,,. AcHN,, O" tBu 0 Ph 1G. (±)-(2R,3R,5R, 1'S)-1 -Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(t-butyl dimethylsilyloxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R, 1 'R)- and (±)-(2R,3R,5R, 1 'S)-l-benzyl-2-(1 amino-3-ethyl)pentyl-3-(t-butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (110 mg, 0.21 mmole) and acetic anhydride (214 mg, 2.1 mmole) in 10 mL of dichloromethane was stirred for 1 hour. The solvent and excess acetic anhydride were removed in vacuo. The residue was purified by chromatography on silica gel using 30% ethyl acetate/hexanes to provide the title compound as a white solid (yield: 85 mg, 72%). 1 H NMR (CDCI 3 ) 8 7.28 (m, 5H), 5.14 (d, J=14Hz, 1H), 4.36 (m, 1H), 3.95 (m, 2H), 3.62 (m, 1H), 3.52 (m, 1H), 3.45 (m, 1H), 2.98 (m, 1H), 1.98 (s, 3H), 1.60 (m, 2H), 1.43 (s, 9H), 1.20-1.40 (m, 7H), 0.88 (s, 9H), 0.80 (m, 6H), 0.04 (s, 6H) MS (M+H) = 561. HO-, AcHN,, OtBu Ph 1H. (i)-(2R,3R,5R, 1'S)-1 -Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(hvd roxy methyl)-pyrrolidine-5-carboxvlic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R,1'S)-l-benzyl-2-(1-amino-3-ethyl)pentyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (85 mg, -85- WO 99/54299 PCT/US99/07945 0.15 mmole) in dry THF (5 mL) was prepared and maintained at room tempera ture under a nitrogen atmosphere. Tetrabutylammonium fluoride (1M in THF, 0.23 mL) was added slowly to the solution. The reaction mixture was stirred for 1 hour. The solvent was removed in vacuo and the residue purified by chromatography on silica gel using 30-50% ethyl acetate/hexanes to provide the title compound as a white foam (yield: 41 mg, 61%). 1 H NMR (CDCl 3 ) 6 7.20-7.35 (m, 5H), 5.20 (d, J=14Hz, 1H), 4.28 (m 1H), 4.93 (m, 2H), 3.65 (m, 2H), 3.50 (m, 1H), 3.23 (m, 2H), 2.22 (m, 2H), 1.98 (s, 3H), 1.62 (m, 1H), 1.43 (s, 9H), 1.15-1.40 (m, 7H), 0.80 (m, 6H) MS (M+H) = 447. MeO-,,. AcHN,, OtBu Ph 11. (±)-(2R, 3R,5R, 1 'S)-1-Benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-(methoxy methyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A mixture of (±)-(2R,3R,5R,1'S)-l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 (hydroxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (40 mg, 0.09 mmole) and silver oxide (200 mg, 0.90 mmole) in 3 mL of iodomethane was heated at reflux for three hours. The reaction was cooled, filtered, and the solvent was removed in vacuo, to provide the title product as a crude oil. MS (M+H) = 461. -86- WO 99/54299 PCT/US99/07945 MeO-, AcHN,, OtBu 'N HH 1 J. (±)-(2R, 3R,5R, 1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(methoxymethyl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. A mixture of the crude (±)-(2R,3R,5R,1'S)-l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-(methoxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (32 mg, 0.07 mmole), prepared according to the method described in Example 11, and ammonium formate (130 mg, 2.1 mmole) in ethanol (5 mL) was heated at reflux in the presence of a catalytic amount of 10% palladium, on activated carbon, for 1.5 hours. The reaction was filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 50% ethyl acetate/hexanes followed by 10% methanol/dichloromethane to provide the title compound as a colorless oil (yield: 16 mg, 47%). MS (M+H)4 = 371. MeO-.. AcH N,, OH HCI O 1 K. (±)-(2R, 3R,5R, 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(methoxymethyl) pyrrolidine-5-carboxylic Acid Hydrochloride. A solution of the (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3 (methoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (15 mg) was dissolved -87- WO 99/54299 PCT/US99/07945 in 6 N HCI in water (1 mL) and stirred at room temperature for 3 hours. The solvent was removed under high vacuum to provide the title compound as a white solid. 1 H NMR(d 6 -DMSO) 5 8.10 (d, J=14Hz, 1H), 4.28 (m, 1H), 4.18 (m,1H), 3.45 (m, 1H), 3.22 (s, 3H), 2.47 (m, 1H), 2.38(m, 1H), 1.90 (m, 1H), 1.88 (s, 3H), 1.15-1.42 (m, 7H), 0.82 (t, J=12.5Hz, 3H), 0.79 (t, J=12.5Hz, 3H) MS (M+H) = 315, (M-H) = 313. Example 2 (±)-(2R,3R,5 R.1 'S)-2-(1-Acetamido-3-ethyl)pentvl-3-methoxvcarbonyl-pyrrolidine 5-carboxylic Acid Hydrochloride. 0 AcHN,, OtBu Ph 2A. (±)-(2R.3R,5R,1 'S)-l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-formyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of oxalyl chloride (0.11 mL, 2M in CH 2
CI
2 ) in 5 mL of anhydrous dichloromethane was prepared and maintained, under a nitrogen atmosphere, at -78 0C. DMSO (32 mg, 0.42 mmole) was added slowly to the solution. The mixture was stirred for 15 minutes and treated with (±)-(2R,3R,5R,1'S)-1-benzyl 2-(1-acetamido-3-ethyl)pentyl-3-hydroxymethyl-pyrrolidine-5-carboxylic acid t butyl ester (38 mg, 0.085 mmole) in 5 mL of dichloromethane. The solution was stirred for 1 hour and triethylamine (86 mg, 0.85 mmole) was added slowly to the reaction mixture. The solution was allowed to warm to room temperature and -88- WO 99/54299 PCT/US99/07945 diluted with dichloromethane. The organic layer was washed with water and brine, dried over MgSO 4 , filtered and concentrated. The residue was purified by chromatography on silica gel using 3% ethyl acetate/hexanes to provide the title compound as a colorless oil (yield: 39 mg, 97%). 1 H NMR (CDCI 3 ) 8 9.68 (d,J=1.0Hz, 1H), 7.28 (m,5H), 5.06 (d, J=14Hz, 1H), 4.38 (m, 1H), 4.10 (m, 1H), 3.75 (m, 2H), 3.45 (m, 1H), 2.62 (m, 1H), 2.20 (m, 2H), 1.98 (s, 3H), 1.42 (s, 9H), 1.25-1.40 (m, 7H), 0.82 (m, 6H) MS (M+H) = 445. O
HO
AcHN,, OtBu Ph 2B. (±)-(2R.,3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-carboxyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of NaCIO 2 (0.16 g) and NaH 2
PO
4
.H
2 0 (0.17g) in water (1 mL) was added to a solution of (±)-(2R,3R,5R,1'S)-l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester (35 mg, 0.079 mmole) and 2-methyl-2-butene (0.5 mL) dissolved in t-BuOH (1.5 mL) and acetonitrile (1.5 mL) at 0 oC. After 1 hour the reaction was quenched with 10% aqueous Na 2
S
2 0 3 and extracted with dichloromethane. The organic layer was washed with water and brine, dried (MgSO 4 ) and concentrated to provide the title product (yield: -30 mg). MS (M+H) = 461. -89- WO 99/54299 PCT/US99/07945 0
CH
3 0" AcHN,, OtBu Ph 2C. (±)-(2R, 3R, 5R, 1'S)-1-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3 methoxycarbonyl-pyrrolidine-5-carboxylic Acid t-Butvl Ester. A solution of Diazald T M (0.5 g, 2.33 mmole) in 5 mL of ether was added slowly to a solution of aqueous KOH (0.5 g in 1 mL of water) and 1 mL of ethanol maintained at 65 oC. Diazomethane was distilled into a receiving flask charged with a solution of (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 carboxyl-pyrrolidine-5-carboxylic acid t-butyl ester (30 mg, 0.065 mmole) in 3 mL of THF. The receiving flask was cooled to 0 oC in an ice/water bath. The condenser was cooled with dry ice/acetone and 3 mL of ether was added the distilling flask until the distillate was colorless. The reaction was stirred for an additional 0.5 hours at 0 OC. The yellowish reaction mixture was quenched with acetic acid (0.1 mL ) and diluted with ethyl acetate. The organic layer was washed with 10% NaHCO 3 and brine, dried with MgSO 4 and concentrated in vacuo. The residue was purified by chromatography on silica gel using 50 % ethyl acetate/hexanes to provide the title compound as a colorless oil (yield: 20 mg, 65%). 1 H NMR (CDCI 3 ) 8 7.25 (m, 5H), 5.10 (d, J=14Hz, 1H), 4.23 (m, 1H), 4.08 (m,1H), 3.85 (m, 1H), 3.72 (m, 1H), 3.69 (s, 3H), 3.40 (m, 1H), 2.75 (m, 1H), 2.33 (m, 1H), 2.15 (m, 1H), 1.98 (s, 3H), 1.42 (s, 9H), 1.20-1.40 (m, 7H), 0.83 (m, 6H) MS (M+H) = 475. -90- WO 99/54299 PCT/US99/07945 0 MeO AcHN,, OtBu H 0 0 2D. (±)-(2R,3R,5R, l'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-methoxycarbonyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. A mixture of (±)-(2R,3R,5R,1'S)-1 -benzyl-2-(1-acetamido-3-ethyl)pentyl-3 methoxycarbonyl-pyrrolidine-5-carboxylic acid t-butyl ester (14 mg, 0.03 mmole) and ammonium formate (0.3 g) in ethanol (1.5 mL) with a catalytic amount of 10% palladium on activated carbon was heated at about 75 oC, for 1 hour. After filtration to remove the catalyst, the solvent was removed in vacuo. The residue was purified by chromatography on silica gel to provide the title compound as a colorless oil (yield: 8.5 mg, 73%). MS (M+H)* = 385. O MeO AcHN,, . OH H 0 O HCI 2E. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-methoxycarbonyl pyrrolidine-5-carboxylic Acid Hydrochloride. A solution of (±)-(2R,3R,5R, 1 'S)-2-(1-acetamido-3-ethyl)pentyl-3-methoxy carbonylpyrrolidine-5-carboxylic acid t-butyl ester (8.5 mg, 0.022 mmole) in 4 N HCI in dioxane (1 mL) was stirred at room temperature for 24 hours. The solvent -91- WO 99/54299 PCT/US99/07945 was removed in vacuo to provide the title compound as an off-white solid (yield 8 mg, 100%). 1 H NMR (d 6 -DMSO) 6 8.02 (d, J=14Hz, 1H), 4.40 (m, 1H), 4.22 (m, 1H), 3.85 (t, J=13Hz, 1H), 3.70 (m, 1H), 3.65 (s, 3H), 3.15 (m, 1H), 2.55 (m, 1H), 2.20 (m, 1H), 1.84 (s, 3H), 1.12-1.42 (m, 7H), 0.82 (t, J=12.5Hz, 3H), 0.68 (t, 3H) MS (M+H) = 329, (M-H) = 327. Example 3 (±)-(2R,3R,5R,1'S)-2-(1 -Acetamido-3-ethyl)Pentyl-3-cyano-pyrrolidine-5 carboxylic Acid Hydrochloride. H._NOH AcHN,, OtBu Ph 3A. (±)-(2R,3R,5R, 1 'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3 (hydroxyiminoformyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound is prepared by reacting a solution of (±) (2R,3R,5R, 1 'S)-1 -benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-formyl-pyrrolidine-5 carboxylic acid t-butyl ester with hydroxylamine hydrochloride and 10% aqueous potassium carbonate in methanol according to the procedure described by Chelucci et al., Tetrahedron: Asymmetry 5:1973 (1994). -92- WO 99/54299 PCT/US99/07945 N C, AcHN,, OtBu Ph 3B. (±)-(2R,3R,5R, 1'S)-1 -Benzyl-2-(1 -acetamido-3-ethyl)pentvl-3-cyano pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound is prepared by reacting a solution of (±) (2R,3R,5R, 1 'S)-l -benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(hydroxyiminoformyl) pyrrolidine-5-carboxylic acid t-butyl ester with 1, l'-carbonyldiimidazole in dichloromethane according to the procedure described by Chelucci et aL., Tetrahedron: Asymmetry 5:1973 (1994). N C, AcHN,, OtBu N ",f H 3C. (±)-(2R,3R,5R, 1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-cvano-pyrrolidine-5 carboxylic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 1J, substituting (±)-(2R,3R,5R, 1 'S)-1 -benzyl-2-(1-acetamido-3 ethyl)pentyl-3-cyano-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R,1'S)-l1-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. -93- WO 99/54299 PCT/US99/07945 NC, AcHN,,,H N1 HH HCI 3D. (±)-(2R.,3R,5R, l'S)-2-(1-Acetamido-3-ethyl)pentyl-3-cyano-pyrrolidine-5 carboxylic Acid Hydrochloride. The title compound is prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R, 1 'S)-2-(1-acetamido-3-ethyl)pentyl-3 cyano-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2 (1-acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5-carboxylic acid t butyl ester. Example 4 (±)-(2R,3R,5R, 1 'S,)-2-(1 -Acetamido-3-ethyl)pentyl-3-propionyl-pyrrolidine-5 carboxylic Acid Hydrochloride. \OH AcHN,, OtBu Ph 4A. (±)-(2R,3R,5R,1'S,1"R)- and (±)-(2R,3R,5R,1'S,1"S)-1-Benzyl-2-(1 acetamido-3-ethyl)pentyl-3-(1-hydroxv)propyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. Ethyl magnesium bromide (0.070mL, 3M in ether) was added to a solution of (±)-(2R,3R, 5 R,1'S)-l1-benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-formyl -94- WO 99/54299 PCT/US99/07945 pyrrolidine-5-carboxylic acid t-butyl ester (18 mg, 0.041 mmole) in 3 mL of tetrahydrofuran. The reaction mixture was maintained at 0 0C, and stirred for 1hour. The reaction was quenched with aqueous ammonium chloride and partitioned between ethyl acetate and water. The organic layer was dried over MgSO 4 , filtered and concentrated to provide the title product (crude yield: 20mg, 100%). MS (M+H)* = 475. AcHN/,, OtBu Ph 4B. (±)-(2R,3R,5R,1'S.)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-propionyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 2A, substituting (±)-(2R,3R,5R,1'S,1"R)- and (±)-(2R,3R,5R,1'S, 1"S)-1 benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(1-hydroxy)propyl-pyrrolid ine-5-carboxylic acid t-butyl ester, 20 mg 0.041 mmole), in place of (±)-(2R,3R,5R,1'S)-1-benzyl-2 (1-acetamido-3-ethyl)pentyl-3-hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 11 mg, 56%). MS (M+H)* = 473. -95- WO 99/54299 PCT/US99/07945 40 AcHN;, OtBu 7 HN 0 4C. (±)-(2R,3R,5R, 1'S,)-2-(1-Acetamido-3-ethyl)pentyl-3-propionyl-pyrrolidine 5-carboxylic Acid t-Butyl Ester. A mixture of (±)-(2R,3R,5R, 1 'S,)-l -benzyl-2-(1 -acetamido-3-ethyl)pentyl-3 propionyl-pyrrolidine-5-carboxylic acid t-butyl ester (11 mg, 0.023 mmole), ammonium formate (250 mg) and palladium (15 mg, 10% on carbon) in ethanol (1.5 mL) was heated at 70 OC for 20 minutes. The reaction was filtered, to remove the catalyst and concentrated. The residue was purified by chromatography on silica gel using 5% methanol/chloroform to provide the title compound (yield: 8.5 mg, 95%). MS (M+H) = 383. AcH N,, H N "' H 0 HCI 4D. (±)-(2 R. 3R,5R, 1 'S,)-2-(1 -Acetamido-3-ethyl)pentyl-3-propionyl-pyrrolidine 5-carboxylic Acid Hydrochloride. A solution of (±)-(2R,3R,5R, 1 'S,)-2-(1-acetamido-3-ethyl)pentyl-3 propionyl-pyrrolidine-5-carboxylic acid t-butyl ester (8 mg) was dissolved in 4 N HCI in dioxane (1 mL) and stirred at room temperature for 24 hours. The reaction -96- WO 99/54299 PCT/US99/07945 was concentrated in vacuo to provide the title compound as an off white solid (yield: 8 mg, 100%). 1 H NMR (DMSO-d 6 ) 8 8.03 (d, J=14Hz, 1H), 4.41 (m, 1H), 4.20 (m, 1H), 3.92 (m, 1H), 3.68 (m, 1H), 3.46 (m, 1H), 2.65 (m, 2H), 2.00 (m, 1H)1.84 (s, 3H), 1.10-1.35 (m, 9H), 0.95 (t, J=Hz,3H), 0.81 (t, J=12.5Hz, 3H), 0.75 (t, J=12.5Hz, 3H) MS: (M-H)- = 325, (M+35) = 361, (2M-H) = 651; (M+H)* = 327, (2M+1) = 653, (2M+Na) + = 675. Example 5 (±)-(2R, 3R, 5R, 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(N-methylcarbamoyl) pyrrolidine-5-carboxylic Acid Hydrochloride. O CH3HN
-
0 AcHN,, - OtBu Ph 5A. (±)-(2R,3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(N-methyl carbamoyl)pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 carboxyl-pyrrolidine-5-carboxylic acid t-butyl ester (0.175 mmole) and triethylamine (18 mg, 0.175 mmole) in 10 mL THF was cooled in an ice-bath. Isobutylchioroformate (24 mg, 0.175 mmole) was added and stirred for 30 min. Then methylamine (2.0 M in THF, 0.35 inmL, 0.70 mmole) was added. The mixture was stirred while allowed to warm up to room temperature overnight. The reaction was then diluted with ethyl acetate. The organic layer was washed with -97- WO 99/54299 PCT/US99/07945 water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 5% methanol / methylene chloride to provide the title compound, as an oil (yield: 17.2 mg, 21%). MS: (M+H)*= 474 0
CH
3
HN
AcHN,, OtBu I HH 0 O 5B. (±)-(2R,3R,5R,1 'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(N-methylcarbamoyl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1 J, substituting (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1 -acetamido-3 ethyl)pentyl-3-(N-methylcarbamoyl)pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxy methyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 13 mg, 94%). MS: (M+H)*= 384 0
CH
3
H
N AcHN,, OH HCI -98- WO 99/54299 PCT/US99/07945 5C. (±)-(2R. 3R,5R, 1'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(N-methylcarbamoyl) pyrrolidine-5-carboxylic Acid Hydrochloride. The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3 ethyl)pentyl-3-(N-methylcarbamoyl)pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-(methoxymethyl) pyrrolidine-5-carboxylic acid t-butyl ester. 'H NMR (D 2 0): 6 4.43 (t, J=10Hz, 1H), 4.36 (m, 1H), 4.09 (dd, 1H), 3.08 (q, J=10Hz, 1H), 2.75 (m, 4H), 2.25 (m, 4H), 2.02 (s, 3H), 1.5-1.15 (br, 7H), 0.80 (m, 6H). MS: (M+H) = 328. Example 6 (±)-(2R,3R,5R. 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(N-aminocarbamoyl) pyrrolidine-5-carboxylic Acid Hydrochloride. O BocHNHN AcHN,, OtBu Ph 6A. (±)-(2R,3R,5R, 1'S)-1 -Benzyl-2-(1-acetamido-3-ethyl)pentvl-3-(N-(t butoxycarbonyl)aminocarbamoyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 carboxyl-pyrrolidine-5-carboxylic acid t-butyl ester (60 mg, 0.13 mmole), t-butyl carbazate (21 mg, 0.16 mmole), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC, 31mg, 0.16 mmole) and 1-hydroxybenzotriazole (9 mg, -99- WO 99/54299 PCT/US99/07945 0.065 mmole) in 3 mL anhydrous THF was stirred at room temperature for 6 hours. The reaction was then diluted with ethyl acetate. The organic layer was washed with water and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 2% methanol/methylene chloride to provide the title compound, as an oil (yield: 45.6 mg, 61%). MS: (M+H) = 575 0O BOC-HNHN N AcHN/,H
O
B
u Iy0 6B. (±)-(2R,3R,5R,1 'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(N-(t butoxycarbonyl)aminocarbamoyl)pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1J, substituting (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1 acetamido-3-ethyl)pentyl-3-(N-(t-butoxycarbonyl)aminocarbamoyl)pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S))-1-benzyl-2-(1 acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 28 mg, 75%). MS: (M+H) = 484 -100- WO 99/54299 PCT/US99/07945 0
NH
2 HN AcHN,, ,OH HCI 6C. (±)-(2R,3R,5R, 1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(N-aminocarbamoyl) pyrrolidine-5-carboxylic Acid Hydrochloride. The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-(N (t-butoxycarbonyl)aminocarbamoyl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1'S)-2-(1-acetamido-3-ethyl)pentyl-3-(methoxymethyl) pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (D 2 0): 6 4.32 (m, 2H), 4.18 (dd, 1H), 3.14 (q, J=8.4Hz, 1H), 2.75 (m, 1H), 2.26 (m, 1H), 2.01 (s, 3H), 1.50-1.15 (m, 7H), 0.80 (q, J=7.5Hz, 6H) MS: (M+H) = 329 -101- WO 99/54299 PCT/US99/07945 Example 7 (±)-(2R,3R,5R,1'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-ethoxycarbonyl-pyrrolidine-5 carboxylic Acid Hydrochloride. 0 EtO.-0 AcHN,, OtBu Ph 7A. (±)-(2R,3R,5R,1'S)-l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3 ethoxycarbonyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 carboxyl-pyrrolidine-5-carboxylic acid t-butyl ester (42 mg, 0.091 mmole), ethanol (0.5 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC, 36 mg, 0.188 mmole) and 1-hydroxybenzotriazole (7 mg, 0.05 mmole) in 2 mL anhydrous THF was stirred at room temperature overnight. The reaction was then diluted with ethyl acetate. The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 2% methanol/methylene chloride to provide the title compound, as an oil (yield: 36 mg, 33%). 1 H NMR (CDCl 3 ): 8 7.50-7.20 (br, 5H), 5.12 (d, J=9Hz, 1H), 4.60-4.30 (br, 2H), 4.14 (q, J=6Hz, 2H), 4.08 (m, 1H), 3.85 (br, 1H), 3.72 (m, 1H), 3.40 (m, 1H), 2.75 (m, 1H), 2.32 (m, 1H), 1.97 (s, 3H), 1.40 (s, 9H), 1.37 (t, J=6Hz, 3H), 1.20 1.50 (m, 7H), 0.83 (m, 6H). Mass spectrum: (M+H)P = 489 -102- WO 99/54299 PCT/US99/07945 0 EtO-/ AcHN/,, OtBu H 0 0 7B. (±)-(2R,3R,5R, l'S)-2-(1-Acetamido-3-ethyl)pentyl-3-ethoxycarbonyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1 J, substituting (±)-(2R,3R,5R, 1 'S)-1 -benzyl-2-(1-acetamido-3-ethyl) pentyl-3-ethoxycarbonyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R, 1 'S)-1 -benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. Mass spectrum: (M+H) = 399. O EtO AcHN,, OH N H 0 HCI 7C. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-ethoxycarbonyl pyrrolidine-5-carboxylic Acid Hydrochloride. The title compound was prepared according to the method described in Example 2E, substituting (±)-(2R,3R,5R, 1 'S)-2-(1-acetamido-3 ethyl)pentyl-3-ethoxycarbonyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (+)-(2R,3R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3-methoxycarbonyl-pyrrolidine 5-carboxylic acid t-butyl ester. -103- WO 99/54299 PCT/US99/07945 1 H NMR (D 2 0): 6 4.35 (m, 1H), 4.20 (q, J=7.5Hz, 2H), 3.87-3.55 (m, 2H), 3.20 (q, J=7.5Hz, 1H), 2.67 (m, 1H), 2.42 (m, 1H), 2.02 (s, 3H), 1.24 (t, J=7.5Hz, 3H), 1.54-1.15 (m, 7H), 0.82 (m, 6 H). Mass spectrum: (M+H)* = 343, (M-H) = 341. Example 8 (+)-(2R,3R, 5R, 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-acetyl-pyrrolidine-5 carboxylic Acid Hydrochloride. OH AcHN,, . OtBu Ph 8A. (±)-(2R,3R,5R,1 'S,1"R)- and (±)-(2R,3R,5R, I'S,1"S)-I-Benzyl-2-(1 acetamido-3-ethyl)pentyl-3-(1-hydroxy)ethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 4A substituting methyl magnesium bromide in place of ethyl magnesium bromide. -104- WO 99/54299 PCT/US99/07945 0 AcHN,, OtBu Ph 8B (±)-(2R, 3 R,5R, 1 'S)-1 -Benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-acetyI pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 2A, substituting (±)-(2R,3R,5R,1'S,1"R)- and (±)-(2R,3R,5R,1'S,1"S)-1 benzyl-2-(l-acetamid o-3-ethyl)pentyl-3-(1-hydroxy)ethyl-pyrrolidine-5-carboxylic acid t-butyl ester, prepared according to the procedure described in Example 8A, in place of (±)-(2R,3R,5R, 1'S)-l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-hydroxy methyl-pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR(CDCl 3 ) 8 5.00(d, J=9.7Hz, 1H), 3.94(m, 2H), 3.68(m, 1H), 3.55(m, 1H), 2.64(m, 1H), 2.32(m, 1H), 2.29(s, 3H),2.20(m, 1H), 1.94(s, 3H), 1.43(s, 9H), 1.15 11.35(m, 7H), 0.80(m, 6H). MS: (M+H)+=459 -105- WO 99/54299 PCT/US99/07945 0 AcHN,, OtBu 'N "If H1 O 8C. (±)-(2R,3R,5R, l'S)-2-(1-Acetamido-3-ethyl)pentyl-3-acetyI-pyrrolidine-5 carboxylic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 4C, substituting (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-acetyl -pyrrolidine-5-carboxylic Acid t-butyl ester., prepared according to the procedure described in Example 8B, in place of (±) (2R,3R,5R, 1 'S)-1 -benzyl-2-(1-acetamido-3-ethyl)pentyl-3-propionyl -pyrrolidine-5 carboxylic acid t-butyl ester. MS: (M+H)"=369 0 AcHN,, OH H 0 S HCI 8D. (±)-(2R,3R,5R, 1 'S)-2-(1-Acetamido-3-ethyl)pentyl-3-acetyl -pyrrolidine-5 carboxylic Acid Hydrochloride. The title compound is prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3 acetyl-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2-(1 -106- WO 99/54299 PCT/US99/07945 acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester. 'H NMR(DMSO-d 6 ) 6 8.20(m, 1H), 4.35(m, 1H), 4.15(m, 1H), 4.03(m, 1H), 2.43(m, 1H), 2.03(m, 1H), 1.91(s, 3H), 1.77(s, 3H), 1.55(m, 1H), 1.46(m, 1H), 1.35(m, 2H), 1.12(m, 4H), 0.84(m, 3H), 0.79(m, 3H) MS: (M+H)*=314, (M-H)- =312 Example 9 (±)-(2S,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-amino-pyrrolidine-5 carboxylic Acid Dihydrochloride. O HO OtBu N 'i Ko Ph 9A. (±)-(2S,3R,5R)- and (±)-(2S,3S,5R)-1l-Benzyl-2-vinyl-3-carboxyl pyrrolidine-5-carboxvlic Acid t-Butyl Ester. A solution of (±)-(2S,3R,5R)- and (±)-(2S,3S,5R)-1l-benzyl-2-vinyl-3-formyl pyrrolidine-5-carboxylic acid t-butyl ester (10 g, 31.7 mmole) (8:1 ratio), in 39 mL of ethanol was prepared. The solution was treated with a suspension of silver oxide (8.83 g, 38.1 mmole) and potassium hydroxide (10.86 g, 194 mmole) in 65 mL of water. The reaction was stirred at room temperature for 1 hour and filtered through a pad of Celite®. The ethanol was removed in vacuo. The aqueous solution was acidified with acetic acid to about pH 4. The acidic solution was extracted with ethyl acetate. The organic layer was washed with brine, dried over Na 2
SO
4 , filtered and concentrated to provide the title compound as a brownish oil -107- WO 99/54299 PCT/US99/07945 (crude yield: 8.2 g, 77%). The crude acid was used for the next step without further purification. MS (M+H) = 332, (M-H) = 330. CbzHN, ., OtBu O'B Ph 9B. (±)-(2S,3R,5R)-l1-Benzyl-2-vinyl-3-benzyloxycarbonviamino-pyrrolidine-5 carboxylic Acid t-Butvl Ester. A mixture of (±)-(2S,3R,5R)- and (±)-(2S,3S,5R)-1l-benzyl-2-vinyl-3 carboxyl-pyrrolidine-5-carboxylic acid t-butyl ester (1.0 g, 3.02 mmole), diphenylphosphoryl azide (0.83 g, 3.32 mmole), benzyl alcohol (0.36 g, 4.53 mmole) and triethylamine (0.32 g, 3.32 mmole) in 30 mL of toluene was heated at reflux for 16 hours. The solvent was evaporated and the residue was purified by chromatography on silica gel using 10% ethyl acetate/hexanes to provide the title compound as a colorless oil (yield: 0.86 g, 65%). 1 H NMR (CDCI 3 ) 6 7.20-7.40(m, 10H), 5.70(m, 2H), 5.10-5.23(m, 3H), 4.10(m, 1H), 3.85(m, 1H), 3.62(m, 1H), 3.45(m, 2H), 2.50(m, 1H), 1.70(m, 1H), 1.41(s, 9H). MS (M+H) = 437. -108- WO 99/54299 PCT/US99/07945 CbzHN, H , OtBu 0 0 Ph 9C. (±)-(2R,3R,5R)-l1-Benzyl-2-formyl-3-benzyloxycarbonylamino-pyrrolidine-5 carboxylic Acid t-Butyl Ester. Osmium tetroxide (3 crystals) was added to a stirred solution of the (+) (2S,3R,5R)-1l-benzyl-2-vinyl-3-benzyloxycarbonylamino-pyrrolidine-5-carboxylic acid t-butyl ester (1.10 g, 2.52 mmole), N-methylmorpholine N-oxide (0.95 g, 8.07 mmole), in 27 mL of acetone/water (8:1), maintained at room temperature. After 6 hours, 10% aqueous Na 2 S20 3 was added and stirring continued for an additional 15 minutes. The reaction was extracted with dichloromethane and the organic layer was concentrated to provide the crude diol intermediate. The diol product was used in the next step without additional purification. MS (M+H) = 471. Sodium periodate (1.0 g, 4.52 mmole) was added in portions to a stirred solution of the crude diol (-1.25 g, 2.66 mmole) in 21 mL of THF/water (6:1). The reaction was stirred for 1 hour then diluted with ethyl acetate. The organic layer was washed with water and brine, dried over MgSO 4 , filtered and concentrated. The residue was purified by chromatography on silica gel using 15% ethyl acetate/hexanes to provide the title compound as a colorless oil (yield: 0.66 g, 60%). 1 H NMR (CDCl 3 ) 5 9.44(d, J=1.2Hz, 1H), 7.20-7.40(m, 10H), 5.98(d, J=14Hz, 1H), 5.10(m, 2H), 4.45(m, 1H), 3.90(m, 2H), 3.70(m, 1H), 3.60(m, 1H), 2.43(m, 1H), 1.70(m, 1H), 1.45(s, 9H), MS (M+H) + = 439. -109- WO 99/54299 PCT/US99/07945 9D. (±)-(2R,3R,5R,1'R)- and (±)-(2R,3R,5R,1'S)-1-Benzyl-2-(1-hydroxy-3 ethyl)pentyl-3-benzyloxycarbonylamino-pyrrolidine-5-carboxylic Acid t-Butyl Ester. 1-Bromo-2-ethylbutane (1.7 g, 10.3 mmole) was added a solution of dibromoethane (3 drops) in 15 mL of dry THF, under argon, in a flask charged with magnesium (0.25 g, 10.3 mmole). The reaction mixture was heated at reflux for 45 minutes, until most of the magnesium reacted. The solution was allowed to cool to room temperature and transferred via cannula to a suspension of CuBr-SMe 2 (2.12 g, 10.3 mmole) in 15 mL of dry THF, maintained under argon, at -10 0C. The mixture was stirred for 0.5 hours until the solution turned dark. A solution of (±)-(2R,3R,5R)-1l-benzyl-2-formyl-3-benzyloxycarbonylamino pyrrolidine-5-carboxylic acid t-butyl ester (0.45 g, 1.03 mmole) in 10 mL of THF was added dropwise and stirred for 1.5 hours, while maintaining the temperature at 0 oC. The reaction was quenched with aqueous ammonium chloride, diluted with ethyl acetate, washed with water and brine, dried over MgSO 4 , filtered and concentrated. The residue was purified by chromatography on silica gel using 10% ethyl acetate/hexanes to provide alcohol adducts as a pale yellow oil (yield: 160 mg, 30%). 1 H NMR (CDCI3) 8 7.20-7.40(m, 10H), 6.10(d, J=14Hz, 1H), 5.10(m, 2H), 4.22(m, 1H), 4.01(m, 1H), 3.71(m, 1H), 3.65(m, 2H), 3.55(m, 1H), 3.20(m, 1H), 2.00-2.30(m, 2H), 1.45(s, 9H), 1.15-1.40(m, 7H), 0.84(m, 6H) MS (M+H)* = 525. -110- WO 99/54299 PCT/US99/07945 CbzHN,. O OtBu 0 h 9E. (±)-(2R,3R, 5R)-1 -Benzyl-2-(1 -oxo-3-ethyl)pentvl-3 benzyloxvcarbonylamino-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of oxalyl chloride (0.29 ml, 2 M in CH 2 Cl 2 ) in 5 mL of dry dichloromethane was prepared and maintained under a nitrogen atmosphere at 78 OC. DMSO (90 mg, 1.14 mmole) was added to the solution. The mixture was stirred for 15 minutes. The alcohol adduct, prepared above, (150 mg, 0.286 mmole), in 5 mL of dichloromethane, was added dropwise to the cold (-78 OC) reaction mixture. The solution was stirred, at -78 oC, for 1 hour. Triethylamine (250 mg, 2.29 mmole) was added slowly. The reaction was allowed to slowly warm to room temperature and then diluted with dichloromethane. The organic layer was washed with water and brine, dried over MgSO 4 , filtered and concentrated. The residue was purified by chromatography on silica gel using 5% ethyl acetate/hexanes to provide the title compound (yield: 100 mg, 67%). 1 H NMR (CDCl 3 ) 67.35(m, 10H), 5.10(m, 2H), 4.28(m, 1H), 3.95(m, 2H), 2.60(m, 1H), 2.40(m, 1H), 2.03(m, 1H), 1.70(m, 2H), 1.45(s, 9H), 1.10-1.30(m, 7H), 0.70(m, 6H) MS (M+H)* = 523. -111- WO 99/54299 PCT/US99/07945 CbzHN,
H
2 N .,OtBu Ph 9F. (±)-(2S,3R,5R,1'R)- and (±)-(2S,3R,5R,1'S)-1-Benzyl-2-(1l-amino-3 ethyl)pentyl-3-benzyloxycarbonylamino-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A mixture of (±)-(2R,3R,5R)-l-benzyl-2-(1-oxo-3-ethyl)pentyl-3-benzyloxy carbonylamino-pyrrolidine-5-carboxylic acid t-butyl ester (90 mg, 0.172 mmole), ammonium acetate (400 mg, 5.17 mmole) and sodium cyanoborohydride (65 mg, 1.03 mmole) in 5 mL in methanol was heated at reflux for 18 hours. Additional portions of ammonium acetate and sodium cyanoborohydride were added and heating continued for an additional 2 hours. The reaction was quenched with 1 N sodium hydroxide, and diluted with dichloromethane. The organic layer was washed with water and brine, dried over MgSO 4 , filtered and concentrated. The residue was purified by chromatography on silica gel using 1:1 ethyl acetate/hexanes followed by 5% methanol/dichloromethane to provide the title compounds. (yield: 58 mg, 64%) MS (M+H) = 524. -112- WO 99/54299 PCT/US99/07945 CbzHN, AcHN OtBu Ph 9G. (±)-(2S,3R,5R,1'R)- and (±)-(2S,3R,5R,1'S)-l-Benzyl-2-(1-acetamido-3 ethyl)pentyl-3-benzyloxycarbonylamino-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2S,3R,5R, 1'R)- and (±)-(2S,3R,5R, 1'S)-1 -benzyl-2-(1 amino-3-ethyl)pentyl-3-benzyloxycarbonylamino-pyrrolidine-5-carboxylic acid t butyl ester (50 mg, 0.096 mmole) and acetic anhydride (117 mg, 1.15 mmole) in 5 mL of dichloromethane was stirred for 1 hour at room temperature. The solvent was evaporated in vacuo and the residue purified by chromatography on silica gel using 30-50% ethyl acetate/hexanes to provide the title compound as a colorless oil (yield: 51 mg, 97%). 1 H NMR (CDCI 3 ) 8 7.72-7.35(m, 10H), 5.82(d, J=14Hz, 1H), 5.10(m, 2H), 4.38(m, 1H), 4.15(m, 2H), 3.63(m, 1H), 3.38(m, 1H), 3.10(m, 1H),2.15(m, 1H), 2.00(s, 3H), 1.65(m, 1H), 1.42(s, 9H), 1.20-1.35(m, 7H), 0.80(m, 6H) MS (M+H) + = 567. -113- WO 99/54299 PCT/US99/07945
H
2 N, AcHN,, OtBu H 0 9H. (±)-(2S,3R,5R, 1 'S)-2-(1-Acetamido-3-ethyl)pentyl-3-amino-pyrrolidine-5 carboxylic Acid t-Butyl Ester. A solution of (±)-(2S,3R,5R,1'R)- and (±)-(2S,3R,5R,1'S)-l-benzyl-2-(1 acetamido-3-ethyl)pentyl-3-benzyloxycarbonylamino-pyrrolidine-5-carboxylic acid t-butyl ester (49 mg, 0.087 mmole), ammonium formate (150 mg, 0.22 mmole) and 10% palladium on activated carbon in ethanol (5 mL) was heated at 80 OC for 45 minutes. After filtration to remove the catalyst, the solvent was removed. The residue was purified by chromatography on silica gel using 5-10% methanol/dichloromethane to furnish the diastereomers, (±)-(2S,3R,5R,1'S) (19 mg) and (±)-(2S,3R,5R,1'R) (8.6 mg) of 2-(1-acetamido-3-ethyl)pentyl-3-amino pyrrolidine-5-carboxylic acid t-butyl ester. 'H NMR (CDCl 3 ) 6 6.00(d, J=14Hz, 1H), 3.90(m, 1H), 3.73(m, 1H), 3.49(m, 1H), 3.10(m, 1H), 2.48(m, 1H), 2.03(s, 3H), 1.82(m, 1H), 1.48(s, 9H), 1.15 1.42(m, 7H), 0.85(m, 6H) MS (M+H)" = 342. -114- WO 99/54299 PCT/US99/07945 H2NA 'N AcHN/,, ,OH H0 HCI 91. (±)-(2S,3R,5R, 1'S)-2-(1 -Acetamido-3-ethyl)pentvl-3-amino-pyrrolidine-5 carboxylic Acid Dihydrochloride. A solution of (±)-(2S,3R,5R, 1 'S)-2-(l1-acetamido-3-ethyl)pentyl-3-amino pyrrolidine-5-carboxylic acid t-butyl ester (17 mg, 0.050 mmole) in 1 mL of 6 N HCI was stirred at room temperature for 3 hours. The solvent was removed under high vacuum to provide the title compound as a white solid (yield: 15 mg, 100%) 1 H NMR (d 6 -DMSO) 8 8.28(bs, 1H), 7.90 (d, J=Hz,1H), 4.71 (d, J=14Hz, 1H), 4.39(m, 1H), 4.10(m, 1H), 3.92(m, 1H), 3.08(m, 1H), 2.64(m, 1H), 2.31(m, 1H), 1.95(m, 1H), 1.88(s, 3H), 1.50(m, 1H), 1.10-1.40(m, 7H), 0.72-0.90(m,6H). MS (M+H)* = 286. -115- WO 99/54299 PCT/US99/07945 Example 10 (±)-(2S, 3R,5R, 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-acetamido-pyrrolidine-5 carboxylic Acid Hydrochloride.
H
2 N, AcHN,, OtBu Ph 10A. (±)-(2S,3R,5R, 1'S)-l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-amino pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2S,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 benzyloxycarbonylamino-pyrrolidine-5-carboxylic acid t-butyl ester (50 mg, 0.88 mmole) was stirred with 10% palladium on carbon (5 mg) in 50 mL of ethyl acetate under 1 atmosphere of hydrogen for 45 minutes. The reaction was filtered and concentrated to provide the title compound as an oil (crude yield: 35 mg, 92%). MS (M+H) = 431. AcHN, AcHN, OtBu N 0 Ph 10B. (±)-(2S,3R,5R,1'S)-l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-acetamido pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2S,3R,5R, 1'S)-1 -benzyl-2-(1-acetamido-3-ethyl)pentyl-3 amino-pyrrolidine-5-carboxylic acid t-butyl ester (35 mg, 0.080 mmole) was -116- WO 99/54299 PCT/US99/07945 reacted with acetic anhydride (0.05 mL) in 8 mL of dichloromethane for 1 hour. The reaction was concentrated and the residue purified by chromatography on silica gel using 50% ethyl acetate/hexanes followed by 3% methanol/dichloromethane to provide the title compound (yield: 30 mg, 80%). 1 H NMR (CDCI3) 57.20-7.35(m, 5H), 6.62(d, J=14Hz, 1H), 5.34(d, J=14Hz, 1H), 4.42(m, 2H), 4.20(m, 1H), 3.68(m, 1H), 3.42(m, 1H), 3.10(m, 1H), 2.18(m, 2H), 2.02(s, 3H), 1.96(s, 3H), 1.45(s, 9H), 1.25-1.42(m, 7H), 0.85(m, 6H). MS (M + H) = 474. AcHN, AcHN,,. ,, OtBu H 0 10C. (±)-(2S,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-acetamido-pyrrolidine 5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1J, substituting (±)-(2S,3R,5R,1'S)-l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-acetamido-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R, 1'S)-l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. The residue was purified by chromatography on silica gel using 5% methanol/dichloromethane to provide the title compound (yield: 11.5 mg, 50%). 1 H NMR (CDCI 3 ) 8 6.20(d, J=14Hz, 1H), 5.94(d, J=14Hz, 1H), 4.24(m, 1H), 4.08(m, 1H), 3.95(m, 1H), 3.75(m, 1H), 3.18(m, 1H), 2.45(m, 1H), 2.02(s, 3H), 1.96(s, 3H), 1.82(m, 1H), 1.49(s, 9H), 1.20-1.42(m, 7H), 0.85(m, 6H). -117- WO 99/54299 PCT/US99/07945 MS (M + H) = 384. AcHN, AcHN,, OH N H 0 HCI 10D (±)-(2S,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-acetamido-pyrrolidine 5-carboxylic Acid Hydrochloride. The title compound was prepared according to the method described in Example 1K, substituting (±)-(2S,3R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3 acetamido-pyrrolidine-5-carboxylic acid t-butyl ester (11.0 mg, 0.029 mmole) in place of (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-(methoxymethyl) pyrrolidine-5-carboxylic acid t-butyl ester (yield: 11.0 mg, 100%). 1 H NMR(d 6 -DMSO) 6 8.15(d, J=14Hz, 1H), 8.05(d, J=14Hz, 1H), 4.35(m, 1H), 4.28(m, 1H), 4.19(m, 1H), 3.59(m, 1H), 1.90(s, 3H), 1.81(s, 3H), 1.15 1.40(m, 7H), 0.80(m, 6H). MS: (M-H) = 326, (M+35) + = 362; (M+H) + = 328, (M+23) + = 350. -118- WO 99/54299 PCT/US99/07945 Example 11 (±)-(2S,3R,5R, 1 'S)-2-(l1-Acetamido-3-ethyl)pentyl-3-methoxycarbonvylamino pyrrolidine-5-carboxylic Acid Hydrochloride. MeO 2 CHN, AcH N, . OtBu Ph 11A. (±)-(2S,3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxy carbonylamino-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2S,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 amino-pyrrolidine-5-carboxylic acid t-butyl ester is reacted with methyl chloroformate and triethylamine in dichloromethane. The reaction is partitioned between dichloromethane and water. The organic layer is concentrated to provide the title compound. MeO 2 CHN, AcHN,, OtBu tN H H 11B. (±)-(2S,3R.,5R, 1'S)-2-(1-Acetamido-3-ethyl)pentyl-3 methoxycarbonylamino-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 1 J, substituting (±)-(2S,3R,5R,1'S)-1 -benzyl-2-(1-acetamido-3-ethyl) pentyl-3-methoxycarbonylaminopyrrolidine-5-carboxylic acid t-butyl ester in place -119- WO 99/54299 PCT/US99/07945 of (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. MeO 2 CHN, AcHN, OH H 0 HCI 11C. (±)-(2S,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3 methoxycarbonylamino-pyrrolidine-5-carboxylic Acid Hydrochloride. The title compound is prepared according to the method described in Example 1K, substituting (±)-(2S,3R,5R, I'S)-2-(1-acetamido-3-ethyl)pentyl-3 methoxycarbonylamino-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R, 3 R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5 carboxylic acid t-butyl ester. -120- WO 99/54299 PCT/US99/07945 Example 12 (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(imidazol-4-yl)-5-carboxylic Acid Dihydrochloride. O N20 AcHN/,. ., OtBu HN " '0 Ph 12A. (±)-(2R,3R,5R, I'S)-l1-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-diazoacetyl 5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R, 1'S)-l1-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 carboxyl-pyrrolidine-5-carboxylic acid t-butyl ester (405.3 mg, 0.88 mmol) and N methylmorpholine (106 9l, 0.96 mmol) in THF (20 ml) was reacted with isobutyl chloroformate (96 ld, 0.93 mmole) at -10 OC for 30 minutes. To the reaction flask was cannulated a distilled diazomethane solution in ether prepared from the reaction of diazald (2.4 g) in ether (60 ml) with a solution of potassium hydroxide (2.4 g) in ethanol (15 ml) and water (15 ml). The reaction was stirred for 3 hours at room temperature then diluted with ether. The organic layer was washed with brine, dried (Na 2
SO
4 ) and concentrated to give the title compound as a thick oil (430.4 mg). -121- WO 99/54299 PCT/US99/07945 Br 0 o AcHN/,, OBu Ph 12B. (±)-(2R,3R,5R,1'S)-1l-Benzyl-2-(1-Acetamido-3-ethyl)pentyl-3-bromoacetyl 5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R, 1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 diazoacetyl-5-carboxylic acid t-butyl ester (427.4 mg, 0.88 mmol) in dioxane (50 ml) was reacted with hydrobromic acid (0.25 ml, 2.2 mmol) at 0 'C for 0.5 hours. The reaction was quenched with saturated aqueous sodium bicarbonate (25 ml) and concentrated in vacuo. The residual aqueous layer was extracted with dichloromethane (3x50 ml). The combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 5% methanol in dichloromethane to provide the title compound as a white foamy solid (379.3 mg, 80.2 %). MS: (M+H)
+
= 539. -122- WO 99/54299 PCT/US99/07945 /N AcHN,, OtBu . N ."I Ph 12C. (±)-(2R,3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(imidazol-4 yl)-5-carboxylic Acid t-ButvI Ester. (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-bromoacetyl 5-carboxylic acid t-butyl ester (60 mg, 0.112 mmol) was treated with formamidine acetate (120 mg, 1.15 mmol) in liquid ammonia and heated at 45 OC in a sealed tube for 20 h. The reaction was concentrated in vacuo. The residue was treated with aqueous NaHCO 3 and extracted with dichloromethane (5x20 ml). The combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 5% methanol in dichloromethane to provide the title compound as a white solid (21.2 mg, 39.4 %). MS: (M+H) = 483. /N H N ,,,.J., AcHN,, ., OBu
.
H N 0 12D. (±)-(2R,3R,5R, 1 'S) -2-(1 -Acetamido-3-ethyl)pentyl-3-(imidazol-4-yl)-5 carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1 J, substituting (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1-acetamido-3-ethyl) -123- WO 99/54299 PCT/US99/07945 pentyl-3-(imidazol-4-yl)-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxy-methyl pyrrolidine-5-carboxylic acid t-butyl ester (yield: 12.9 mg, 66.2%). 1 H NMR (CDCI 3 ): 8 0.75 -0.81 (m, 6H), 1.17 - 1.42 (m, 7H), 1.47 (s, 9H), 2.03 (s, 3H), 2.66 (m, 1H), 3.50 (m, 1H), 3.73 (m, 1H), 3.86 (m, 1H),4.06 (m, 1H), 7.04 (br s, 1H), 7.86 (br s, 1H). MS: (M+H) = 393. /" N HN N AcHN,, , OH H 2 HCI 12E. (±)-(2R,3R,5R,1'S) -2-(1l-Acetamido-3-ethyl)pentyl-3-(imidazol-4-yl)-5 carboxylic Acid Dihydrochloride. The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3 (imidazol-4-yl)-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2-(1 acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester to provide the title compound solid (yield: 12.0 mg, 96.0%). 1 H NMR (DMSO-d 6 ): 8 0.67 (t, J = 7 Hz, 3H), 0.75 (t, J = 7 Hz, 3H), 1.11 (m, 3H), 1.23 (m, 4H), 1.78 (s, 3H), 2.33 (m, 1H), 2.70 (m, 1H), 3.69 (dt, 1H), 3.95 (dd, 1H), 4.29 (m, 1H), 4.48 (dd, 1H), 7.63 (s, 1H), 8.28 (d, J = 9 Hz, 1H), 9.06 (s, 1H). MS: (M+H) = 337. -124- WO 99/54299 PCT/US99/07945 Example13 (±)-(2R,3R,5R, 1 'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(oxazol-2-yl)-pyrrolidine-5 carboxylic Acid Dihydrochloride. OH O N> H O~ AcHNH OBu 'N Ph 13A. (±)-(2R,3R,5R,1 'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(N-(2 hydroxvethyl)carbamoyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 5A, substituting ethanolamine for N-methylamine hydrochloride. 0 N ' AcHN,, OtBu ~o Ph 13B. (±)-(2R,3R,5R, 1 'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentvyl-3-(oxazolin-2 yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 (N-(2-hydroxyethyl)carbamoyl)-pyrrolidine-5-carboxylic acid t-butyl ester, triethylamine (4 eq.), carbon tetrachloride (3.5 eq.) in acetonitrile is reacted with triphenylphosphine (3.15 eq.) for 16h at room temperature. The reaction is concentrated in vacuo. The residue is partitioned between ethyl acetate and -125- WO 99/54299 PCT/US99/07945 water. The organic layer is washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue is purified by chromatography on silica gel using ethyl acetate/hexanes to provide the title compound. 0 N AcHN,, OtBu 0 Ph 13C. (±)-(2R,3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(oxazol-2-yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 (oxazolin-2-yl)-pyrrolidine-5-carboxylic acid t-butyl ester is reacted with nickel peroxide in cyclohexane according to the method described by Meyer in J. Org. Chem. 1979, 497-501 to provide the title compound. / 0 N J-11 AcHN/, OBu H0 13D. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(oxazol-2-yl) pyrrolidine-5-carboxvlic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 1 J, substituting (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl) pentyl-3-(oxazol-2-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (+) -126- WO 99/54299 PCT/US99/07945 (2R,3R,5R, 'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. /0 N AcHN,, OH H 0 2 HCI 13E. (±)-(2R,3R,5R, 1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(oxazol-2-yl) pyrrolidine-5-carboxylic Acid Dihydrochloride. The title compound is prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R, 1 'S)-2-(1-acetamido-3-ethyl)pentyl-3 (oxazol-2-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (+) (2R,3R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine-5 carboxylic acid t-butyl ester. -127- WO 99/54299 PCT/US99/07945 Examplel4 (±)-(2S,3R,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(N-methylamino)pyrrolidine 5-carboxylic Acid Dihydrochloride. CH3 CbzN, O-t-Bu N Ph 14A. (±)-(2S,3R,5R)-1l-Benzyl-2-vinyl-3-(N-methyl-N-benzyloxvcarbonylamino) pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2S,3R,5R)-1l-benzyl-2-vinyl-3-benzyloxycarbonylamino pyrrolidine-5-carboxylic acid t-butyl ester (2.08 g, 4.77 mmole) was dissolved in 50 mL of anhydrous DMF and maintained under a nitrogen atmosphere. The solution was treated with sodium hydride (0.32 g, 8 mmole), and stirred at room temperature for 30 minutes. The solution was treated with iodomethane (0.8 ml, 12.85 mmole) and stirred for an additional 1 hour. The reaction was quenched with water and extracted with ethyl acetate. The combined organic layers were concentrated to provide the crude product which was purified by chromatography on silica gel to provide the title compound as an oil (yield: 1.75 g, 81%). 1 H NMR (CDCl 3 ) 8 7.36-7.20 (m, 10OH), 5.75-5.50 (br, 1H), 5.25-5.07 (m, 4H), 4.75-4.50 (br, 1H), 3.97 (d, J=13.5Hz, 1H), 3.75 (m, 1H), 3.61 (d, J=13.5Hz, 1H), 3.50 (m, 1H), 2.93 (s, 3H), 2.45 (m, 1H), 1.75 (m, 1H), 1.46 (s, 9H). MS (M+H)* = 451. -128- WO 99/54299 PCT/US99/07945
CH
3 CbzN, H . O-t-Bu 0 Ko Ph 14B. (±)-(2R,3R,5R)-1l-Benzyl-2-formyl-3-(N-methyl-N-benzyloxycarbonyl amino)pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 9C, substituting (±)-(2S,3R,5R)-1l-benzyl-2-vinyl-3-(N-methyl-N-benzyl oxycarbonylamino)pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2S,3R,5R)-1 -benzyl-2-vinyl-3-benzyloxycarbonylamino-pyrrolidine-5-carboxylic acid t-butyl ester. (Yield: 747 mg, 42%.) MS (M+H) + = 453. CH3 CbzN, O O-t-Bu Ph 14C. (±)-(2R,3R,5R)-1-Benzyl-2-(1-oxo-3-methyl)butyl-3-(N-methyl-N benzyloxvcarbonylamino)pyrrolidine-5-carboxylic Acid t-ButvI Ester. Isobutyl magnesium chloride (2.0 M in ether, 0.68 ml) was added dropwise over about 12 minutes to a solution of (±)-(2R,3R,5R)-1-benzyl-2-formyl-3-(N methyl-N-benzyloxycarbonylamino)pyrrolidine-5-carboxylic acid t-butyl ester (196 mg, 0.43 mmole) in 5 mL of anhydrous THF, maintained at -78 OC. The resulting yellow solution was stirred at -78 OC for 1 hour. The solution was quenched with saturated aqueous ammonium chloride, and extracted with ethyl acetate. The -129- WO 99/54299 PCT/US99/07945 organic layer was concentrated and the crude product was oxidized according to the procedure described in Example 9D. Purification by column chromatography on silica gel, with 10-25% ethyl acetate/hexanes, provided the title compound (yield: 78 mg, 36%). 1 H NMR (CDCI3) 6 7.46-7.25 (m, 10H), 5.09 (br, 2H), 4.90-4.60 (m, 1H), 3.97-3.65 (m, 4H), 3.00 (s, 3H), 2.60 (br, 1H), 2.20-1.80 (m, 3H), 1.46 (s, 9H), 0.80-0.67 (m, 7H). MS (M+H) = 509. CH3 CbzN
H
2 N O-t-Bu Ph 14D. (±)-(2S,3R,5R,1'R)- and (±)-(2S,3R,5R,1'S)-l-Benzyl-2-(1-amino-3 methyl)butyl-3-(N-methyl-N-benzyloxycarbonylamino)pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 9F, substituting (±)-(2R,3R,5R)-1l-benzyl-2-(1-oxo-3-methyl)butyl-3-(N methyl-N-benzyloxycarbonylamino)pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R)-1l-benzyl-2-formyl-3-benzyloxycarbonylamino-pyrrolidine 5-carboxylic acid t-butyl ester. (Yield: 97 mg, 65%.) MS (M+H)+ = 510. -130- WO 99/54299 PCT/US99/07945 CH3 CbzN, AcHN . O-t-Bu Ph 14E. (±)-(2S,3R,5R,1'R)- and (±)-(2S,3R,5R,1'S)-l-Benzyl-2-(1-acetamido-3 methvl)butyl-3-(N-methyl-N-benzyloxycarbonylamino)pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2S,3R,5R, 1 'R)- and (±)-(2S,3R,5R, 1 'S)-l-benzyl-2-(1 amino-3-methyl)butyl-3-(N-methyl-N-benzyloxycarbonylamino)pyrrolidine-5 carboxylic acid t-butyl ester (47 mg, 0.094 mmole) was reacted with acetic anhydride (0.15 mL) in 4 mL of dichloromethane at room temperature for 2 hours. The reaction was concentrated in vacuo to provide the title compound. MS (M+H) = 552. CH3 HN 'N ' HH ~0 14F. (±)-(2S,3R,5R,1'S)-2-(1-Acetamido-3-methyl)butyl-3-(N-methylamino) pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2S,3R,5R,1'R)- and (±)-(2S,3R,5R, 1'S)-1-benzyl-2-(1 acetamido-3-methyl)butyl-3-(N-methyl-N-benzyloxycarbonylamino)pyrrolidine-5 carboxylic acid t-butyl ester (0.094 mmole), palladium (40 mg, 10% on carbon) and ammonium formate (160 mg) in 3 mL of ethanol was heated at reflux for 30 minutes. Additional palladium on carbon (15 mg) and ammonium formate (50 -131- WO 99/54299 PCT/US99/07945 mg) were added. The solution was stirred for an additional 15 minutes and the mixture was then filtered to remove the solids and catalyst. The filtrate was evaporated and the residue purified by chromatography on silica gel 5% methanol/dichloromethane and 1% NH 4 OH to provide (±)-(2S,3R,5R,1'S) (15.4 mg, lower Rf) and (±)-(2S,3R,5R,1'R) (5.4 mg, higher Rf)- 2-(1-acetamido-3 methyl)butyl-3-(N-methylamino)pyrrolidine-5-carboxylic acid t-butyl esters (yield: 20.8 mg, 68%). CH3 H N, AcHN,, .,OH ~O H 0 HCI 14G. (±)-(2S,3R,5R,1'S)-2-(1-Acetamido-3-methyl)butvl-3-(N-methylamino) pyrrolidine-5-carboxylic Acid Dihydrochloride. A solution of (±)-(2S,3R,5R, 1 'S)-2-(1-Acetamido-3-methyl)butyl-3-(N methylamino)pyrrolidine-5-carboxylic acid t-butyl ester (9.4 mg) was stirred with 4 N aqueous HCI (-1.5 mL) for 2 hours. The reaction was concentrated in vacuo to provide the title compound (yield: 10 mg, 100%). 1H NMR (major peaks) (DMSO-d 6 ) 52.57 (s, 3H), 1.90 (s, 3H), 1.47 (m, 3H), 0.91 (d, J=7.5Hz, 3H), 0.83 (d, J=7.5Hz, 3H)
MS:(M+H)
+ = 272. -132- WO 99/54299 PCT/US99/07945 Example15 (±)-(2R,3R,5R, 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(imidazol-2-yl)-pyrrolidine-5 Carboxylic Acid Dihydrochloride. r NH N H AcHN/,, O-t-Bu Ph 15A. (±)-(2R,3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(imidazol-2 yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. Ammonia gas was bubbled slowly through a solution of (+) (2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-formyl-pyrrolidine-5 carboxylic acid t-butyl ester (20 mg, 0.045 mmole) and glyoxal (6.2 uL, 0.054 mmole, 1.2 equiv.) in 5 mL of methanol, maintained at 0 oC, for 5 minutes. After 7 hours at 0 0C, additional glyoxal (10 uL) was added and ammonia was bubbled through the solution for 5 minutes. The reaction was allowed to stir at room temperature for 16 hours. A final addition of glyoxal (10 uL) and ammonia as, described above, followed by reaction at room temperature for an additional 4 hours effected a complete reaction. The reaction was concentrated in vacuo and purified by chromatography on silica gel using 50% ethyl acetate/hexanes, followed by 10% methanol/chloroform to provide the title compound as a solid (yield: 19.9 mg, 91%). 1 H NMR (CDCl3): d 0.67 (t, J=7.2 Hz, 3H), 0.73 (t, J=7.2 Hz, 3H), 1.09 1.32 (m, 7H), 1.41 (s, 9H), 2.00 (m, 1H), 2.09 (s, 3H), 2.79 (m, 1H), 3.29 (m, 1H), 3.66 (dd, J=9.6, 2.7 Hz, 1H), 3.77 (m, 1H), 3.92 (d, J=13.4 Hz, 1H), 4.04 (d, -133- WO 99/54299 PCT/US99/07945 J=13.4 Hz, 1H), 4.22 (dd, 1H), 4.49 (m, 1H), 6.08 (brs, 1H), 7.00 (s, 2H), 7.21 7.34 (m, 5H). MS (M+H) = 483. "NH N- 'if 7AcHN,, O-t-Bu f HH 15B. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)penyl-3-(imidazol-2-yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester A mixture of (±)-(2R,3R,5R, 1 'S)-1 -benzyl-2-(1-acetamido-3-ethyl)pentyl-3 (imidazol-2-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (17 mg, 0.035 mmole), ammonium formate (250 mg) and 10% palladium on carbon (20 mg), in 5 mL of ethanol, was heated at reflux for 15 minutes The reaction was concentrated in vacuo and the residue was purified by chromatography on silica gel using 5% methanol/dichlormethane and 0.25% ammonium hydroxide to provide the title compound as a white solid (yield: 11.3mg, 81.9%). MS (M+H) = 393. -134- WO 99/54299 PCT/US99/07945 SNH AcHN,, . ,, OH N H 2HCI 15C. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(imidazol-2-yl) pyrrolidine-5-Carboxylic Acid Dihydrochloride. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(imidazol-2-yl) pyrrolidine-5-carboxylic acid t-butyl ester (11 mg, 0.028 mmole) was dissolved in 2 mL of 6N HCI and stirred at room temperature for 2 hours. The reaction was concentrated in vacuo to provide the title compound, as an off white solid (yield: 11.3 mg, 100%). 1 H NMR (DMSO-d6): d 0.71 (t, J=7 Hz, 3H), 0.75 (t, J=7 Hz, 3H), 1.09 1.28 (m, 7H), 1.74 (s, 3H), 2.43 (m, 1H), 2.80 (m, 1H), 3.85 (m, 1H), 4.04 (m, 1H), 4.29 (m, 1H), 4.52 (m, 1H), 7.64 (s, 2H), 8.07 (br d, J=9 Hz, 1H). MS (M+H) = 337 and (M-H)- = 335. -135- WO 99/54299 PCT/US99/07945 Example 16 (±)-(2R,3R,5R. 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(N,N-dimethylcarbamoyl) pyrrolidine-5-carboxylic Acid Hydrochloride.
H
3 C O H3C-N AcNH,. Ot-Bu Ph 16A. (±)-(2R,3R,5R,1 'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(N,N dimethylcarbamoyl)pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 5A substituting N,N-dimethylamine in place of N-methylamine (yield: 10 mg, 23%). Mass spectrum: (M+H) = 488.
H
3 C O H3C-N AcNH,,. Ot-Bu 'N1 HH 0 16B. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(N-methylcarbamoyl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1J, substituting (±)-(2R,3R,5R,1'S)-l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-(N,N-dimethylcarbamoyl)pyrrolidine-5-carboxylic acid t-butyl ester -136- WO 99/54299 PCT/US99/07945 in place of (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxy methyl-pyrrolidine-5-carboxylic acid t-butyl ester to provide the title compound (yield: 5.5 mg , 67%). Mass spectrum: (M+H) = 398.
H
3 C 0
H
3
C
N AcHN,, , . OH HH HCI 16C. (±)-(2R,3R,5R,1'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(N,N-dimethyl carbamoyl)pyrrolidine-5-carboxvlic Acid Hydrochloride. The title compound was prepared according to the method described in Example 1 K, substituting (±)-(2R,3R,5R, l'S)-2-(1-acetamido-3-ethyl)pentyl-3 (N,N-dimethylcarbamoyl)pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R, 3 R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine 5-carboxylic acid t-butyl ester. 1 H NMR (major peaks) (D 2 0) d 3.15 (s, 3H), 2.94 (s, 3H), 1.98 (s, 3H), 0.80 (m, 6H) MS (M+H) = 342, (M-H) = 340. -137- WO 99/54299 PCT/US99/07945 Example 17 (±)-(2R, 3R,5 R,1 'S)-2-(1 -Acetamido-3-Ethyl)pentyl-3-cyano-pyrrolidine-5 carboxylic Acid Hydrochloride. N C, O-t-Bu Ph 17A. (±)(2S,3R,5R)-1 -Benzyl-2-vinyl-3-cyano-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2S,3S,5R)-1l-benzyl-2-vinyl-3-formyl-pyrrolidine-5 carboxylic acid t-butyl ester (8:1 ratio) (5g, 15.9 mmole) with hydroxylamine hydrochloride (1.28g, 18.5 mmole) and 10% aqueous potassium carbonate (8 mL) in 20 mL of methanol, according to the procedure described by Chelucci et aL, Tetrahedron: Asymmetry 5:1973 (1994) provided an the intermediate oxime product. The crude oxime, prepared above, was reacted with 1,1' carbonyldiimidazole (3.9, 23.9 mmole) in 50 mL of dichloromethane for 3 hours, at room temperature. The reaction was concentrated in vacuo and chromatographed on silica gel with 2-10% ethyl acetate/hexanes to provide the title compound (yield: 2.5g, 50%). MS (M+H) = 313 -138- WO 99/54299 PCT/US99/07945 NC, H .,, O-t-Bu 0 Ph 17B. (±)-(2R,3R,5R)-1l-Benzyl-2-formyl-3-cyano-pyrrolidine-5-carboxylic Acid t Butyl Ester. The title compound is prepared according to the method described in Example 1D, substituting (±+)(2S,3R,5R)-1l-benzyl-2-vinyl-3-cyano-pyrrolidine-5 carboxylic acid t-butyl esterin place of (±)-(2S,3R,5R)-1l-benzyl-2-vinyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 2.2 g, 80%). MS (M+H) = 315 NC, o . 0-t-Bu H N *"' V o Ph 17C. (±)-(2R,3R,5R)-l-Benzyl-2-(1-oxo-3-ethvl)pentvl-3-cvano-pyrrolidine-5 carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1E, substituting (±)(2S,3R,5R)-1l-benzyl-2-formyl-3-cyano-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R)-1-benzyl-2-formyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield 0.4 g ,27%). MS (M+H) = 399 -139- WO 99/54299 PCT/US99/07945 N C, H2A, O-t-Bu 0 Ph 17D. (±)-(2R,3R,5R, 1 'S)-1 -Benzyl-2-(1-amino-3-ethyl)pentyl-3-cyano-pyrrolidine 5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1F, substituting (±)-(2R,3R,5R)-1l-benzyl-2-(1-oxo-3-ethyl)pentyl-3 cyano-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R)-1 benzyl-2-(1-oxo-3-ethyl)pentyl-3-(t-butyldimethylsilyloxymethyl)-pyrrolidine-5 carboxylic acid t-butyl ester (yield 0.215 g , 50%). MS (M+H) = 400 N C, AcHN,, .O-t-Bu Ph 17E. (±)-(2R,3R,5R, 1 'S)-l-Benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-cyano pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 1G, substituting (±)-(2R,3R,5R,1'S)-1 -benzyl-2-(l1-amino-3-ethyl)pentyl 3-cyano-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'R) and (±)-(2R,3R,5R,1'S)-l-benzyl-2-(1-amino-3-ethyl)pentyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield 0.210 g, 90%). -140- WO 99/54299 PCT/US99/07945 1 H NMR (CDCI 3 ) 5 7.25 (m, 5H), 5.08 (m, 1H), 4.40 (m, 1H), 4.15 (m, 1H), 3.78 (m,1H), 3.48(m, 1H), 2.93 (m, 1H), 2.32 (m, 1H), 2.12 (m, 1H), 2.02 (s, 3H),1.52 (s, 9H), 1.35 (m, 7H), 0.85 (m, 6H) MS: (M+H) = 442. NC, AcHN,, O-t-Bu H0 17F. (±)-(2R, 3R, 5R. 1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-cyano-pyrrolidine-5 carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1J, substituting (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-cyano-pyrrolid i ne-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (CDCI 3 ) 8 5.35 (bs, 1H),4.00 (m, 1H), 3.83 (m, 1H), 3.39 (m, 1H), 3.08 (m, 1H), 2.63 (m, 1H), 2.15 (m, 1 H), 2.05 (s, 3H), 1.48 (s, 9H), 1.20-1.45 (m, 7H), 0.85 (m, 6H) MS: (M+H) =352 -141- WO 99/54299 PCT/US99/07945 N C, AcH N,, 'OH H 0 HCI 17G. (+)-(2R,3R,5R, 1'S)-2-(1-Acetamido-3-ethyl)pentvI-3-cyano-pyrrolidine-5 carboxylic Acid Hydrochloride. The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3 cyano-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2 (1-acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5-carboxylic acid t butyl ester. 1 H NMR (d 6 -DMSO) 8 9.12 (bs, 1H), 8.05 (m, 1H), 4.38 (m, 1H), 4.23 (m, 1H), 3.88 (m, 1H), 3.68 (m, 1H), 3.00 (m, 1H), 2.55 (m, 1H), 2.05 (m, 1H), 1.88 (s, 3H), 1.10-1.40 (m, 7H), 0.80 (m, 6H) MS: (M+H) = 296, (M-H)-= 294, -142- WO 99/54299 PCT/US99/07945 Example 18 (±)-(2R,3S, 5R, 1 'S)-2-( 1 -Acetamido-3-ethyl)pentyl-3-ethyl-pyrrolidine-5-carboxylic Acid Hydrochloride. AcHN. OtBu 'N Ph 18A. (±)-(2R,3S,5R,1 'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentl-3-vinyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. To an ice-cold suspension of methyl triphenylphosphonium bromide (240 mg, 0.67 mmol) in 5 mL THF was added potassium t-butoxide (60 mg, 0.54 mmol) under nitrogen. The color changed immediately to bright yellow. After stirring at room temperature for 1 h, (±)-(2R,3R,5R,1'S)-1-benzyl-2-(1-acetamido 3-ethyl)pentyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester (100 mg, 0.225 mmol) in 5 mL THF was added and stirred at room temperature overnight. Reaction was then quenched with saturated ammonium chloride and extracted with ethyl acetate to give the crude product which was purified by chromatography on silica gel using 30% ethyl acetate/hexanes to provide the title compound, as an oil (yield: 55 mg, 55%). 1 H NMR (CDCl 3 ): 6 7.45-7.20 (m, 5H), 5.94 (ddd, 1H), 5.24 (d, J=12Hz, 1H), 4.98 (d, J=18Hz, 1H), 4.93 (d, J=10.5HZ, 1H), 4.37 (m, 1H), 4.06 (d, J=13.5Hz, 1H), 3.80 (d, J=13.5Hz, 1H), 3.41 (dd, J=9 Hz, J=3Hz, 1H), 3.31 (q, J=13.5Hz, 1H), 2.60 (m, 1H), 2.26 (m, 1H), 2.00 (s, 3H), 1.45 (s, 9H), 1.40-1.25 (m, 7H), 0.82 (m, 6H). MS: (M+H) = 443 -143- WO 99/54299 PCT/US99/07945
CH
3 , AcHN. OtBu 'N H 0 18B. (±)-(2R,3S,5R, 1'S)-2-(1-acetamido-3-ethyl)pentyl-3-ethyl-pyrrolidine-5 carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1 J, substituting (±)-(2R,3S,5R, 1'S)-1l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R,1'S)-l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (CDCI 3 ): 5 5.71 (br, 1H), 4.00 (br, 1H), 3.68 (t, J=8Hz, 1H), 3.10 (m, 1H), 2.38 (m, 1H), 1.98 (s, 3H), 1.87 (m, 1H), 1.47 (s, 9H), 1.55-1.20 (m, 10H), 0.93 (t, J=7.5Hz, 3H), 0.83 (m, 6H). MS: (M+H) = 355
CH
3 -,. AcHN. OH 'N d H 0 HCI 18C. (±)-(2R,3S,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-ethyl-pyrrolidine-5 carboxylic Acid Hydrochloride The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3S,5R, 1 'S)-2-(1-acetamido-3-ethyl)pentyl-3 ethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2 -144- WO 99/54299 PCT/US99/07945 (1-acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5-carboxylic acid t butyl ester. 'H NMR (D 2 0): 8 4.30 (br, 1H), 4.25 (t, J=7.5Hz, 2H), 3.58 (br, 1H), 2.61 (m, 1H), 2.23 (br, 1H), 2.05 (s, 3H), 1.90 (m, 1H), 1.70-1.20 (m, 9H), 0.92 (t, J=7.5Hz, 3H), 0.81 (m, 6H). MS: (M+H) = 299 Example 19 (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-propyl-pyrrolidine-5 carboxylic Acid Hydrochloride. CH3 AcHN. OtBu HN ' o Ph 19A. (±)-(2R,3S,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester and (±)-(2R,3S,5R,1'S)-1l-Benzyl 2-(1-acetamido-3-ethyl)pentyl-3-(trans-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according the method described in Example 18A substituting ethyl triphenylphosphonium bromide for methyl triphenylphosphonium bromide. 1 H NMR (CDCI 3 ) 6 7.24 (m, 5H), 5.59 (m, 1H), 5.36 (dd, J= 11, 7Hz, 1H), 5.28 (bs, 1H), 4.32 (m, 1H), 4.06 (d, J= 12.9Hz, 1H), 3.80 (d, J= 12.9Hz, 1H), 3.42 (dd, J= 8.5, 2.0Hz, 1H), 3.30 (dd, J= 6.1, 3.1Hz, 1H), 2.88 (m, 1H), 2.29 (m, 2H), 2.01 (s, 3H), 1.64 (dd, J= 6.8, 1.7Hz, 3H), 1.44 (s, 9H), 1.30 (m, 7H), 0.81 (m, 6H). -145- WO 99/54299 PCT/US99/07945 MS: (M+H)*= 457, (M+Na)+= 479, (M-H)- = 455.
CH
3 O~ AcHN. OtBu
CH
3 0 19B. (±)-(2R.3S,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-propyl-pyrrolidine-5 carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1J, substituting (±)-(2R,3S,5R, 1 'S)-l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester and (+) (2R,3S,5R, l'S)-1 -benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(trans-propen-1 -yl) pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1-benzyl 2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine-5-carboxylic acid t butyl ester (yield: 3.5 mg, 54%). MS: (M+H) = 369, (M+Na) = 391, (M-H)- = 367.
CH
3 AcHN OH 'N H 0 HCI 19C. (±)-(2R,3S,5R,1'S)-2-( -acetamido-3-ethyl)pentyl-3-propyl-pyrrolidine-5 carboxylic Acid Hydrochloride The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3S,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3 ethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2 -146- WO 99/54299 PCT/US99/07945 (1-acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 3.5 mg, 100%). 1 H NMR (DMSO-d6) 6 8.10 (d, J= 8.3Hz, 1H), 4.24 (m, 1H), 4.17 (m, 1H), 2.43 (m, 1H), 2.19 (m, 1H), 1.89 (s, 3H), 1.70 (m, 1H), 1.50-1.20 (m, 12H), 0.87 (t, J= 6.8Hz, 3H), 0.84 (t, J= 7.0Hz, 3H), 0.79 (t, J= 7.3Hz, 3H). MS: (M+H) = 313, (M+Na)* = 335, (M-H)- = 311. Example 20 (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloride. TBDMSO-, HO ) OtBu HO Ph 20A. (±)-(2R,3R,5R,1 'RS)-1l-Benzvl-2-(1,2-dihydroxy)ethyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. Osmium tetroxide was added to a room temperature solution of (±)-(2S, 3R,5R)-1-benzyl-2-vinyl-3-(t-butyldimethylsilyloxymethyl)-pyrroidine-5 carboxylic acid t-butyl ester (3.5 g, 8.12 mmol) in 60 mL of 8:1 acetone/water and N-methylmorpholine N-oxide (3.0 g, 25.6 mmol). The reaction mixture was stirred at room temperature for 6 hours and quenched with saturated aqueous Na 2
S
2 0 3 . The mixture was stirred for an additional 10 minutes and the solvent removed. The brownish residue was partitioned between dichloromethane and water. The organic layer was dried over MgSO 4 and concentrated in vacuo to provide the intermediate diol as an oil (~3.8g ) which was used without additional purification. -147- WO 99/54299 PCT/US99/07945 MS: (M+H) = 466. TBDMSO-, HO OtBu HO 20B. (±)-(2R,3R,5R,1'RS)-2-(1,2-Dihydroxy)ethyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1 J, substituting (±)-(2R,3R,5R,1'RS)-1l-benzyl-2-(1,2-dihydroxy)ethyl-3 (t-butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (21.5g, 46.2 mmol). in place of (±)-(2R,3R,5R,1'S)-1-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-methoxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester. MS: (M+H)
+
= 367. TBDMSO-, HO OtBu Nfl H 0 o0 HO 20C. (±)-(2R,3R,5R,1'RS)-1l-t-Butoxycarbonyl-2-(1,2-dihydroxy)-ethyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'RS)-2-(1,2-Dihydroxy)ethyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (crude from previous step) was dissolved in 160 mL of 3:1 methanol/water and di-tert-butyl dicarbonate (14.0 g, 64 mmol) was added. The mixture was stirred at room temperature for 72 h. Then solvent was removed and the residue was purified by -148- WO 99/54299 PCT/US99/07945 chromatography on silica gel using 50% ethyl acetate/hexanes to provide the title compound as light yellow solid (yield: 15.4 g, 70 %). 'H NMR (CDCI3): 6 0.03 (s, 3H), 0.05 (s, 3H), 1.37 (s, 9H), .42 (s, 9H), 1.47 (s, 9H), 1.93 (d, 1H), 2.30-2.50 (m, 2H), 3.28 (d, 1H) , 3.66-3.43 (m, 4H), 3.85 (dd, 1H), 4.02-4.52 (m, 1H). MS: (M+H) = 476. TBDMSO-, H OtBu 0 H Boc 20D. (±)-(2R,3R,5R)-1l-t-Butoxycarbonyl-2-formyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R, 1'RS)-1l-t-butoxycarbonyl-2-(1,2 dihydroxy)ethyl-3-(t-butyldimethylsilyloxymethyl)-pyrrolid i ne-5-carboxylic acid t butyl ester (6.0 g, 12.6 mmol) was dissolved in 6:1 tetrahydrofuran (THF)/water (110 mL) and treated with sodium periodate (4.4 g, 20.6 mmol). The mixture was stirred at room temperature for 3 hour and diluted with ethyl acetate, washed with water, dried over MgSO 4 , filtered, and concentrated in vacuo. The residue was purified by chromatography on silica gel using 20% ethyl acetate/hexanes to provide the title compound as a white waxy solid (yield: 4.4 g, 78.6%). 1 H NMR (CDCI 3 ) (mixture of two rotamers): 8 0.05 and 0.06 (two s, 6H), 0.88 and 0.90 (two s, 9H), 1.42 and 1.44 (two s, 9H), 1.47 and 1.48 (two s, 9H), 1.89 -1.99 (m, 1H), 2.37 - 2.43 (m, 2H), 3.54 - 3.67 (m, 2H), 4.02 - 4.34 (m, 2H), 9.43 and 9.53 (two d, 1H). MS: (M+H)*= 444. -149- WO 99/54299 PCT/US99/07945 TBDMSO-, HO OtBu HN' Boc 20E. (±)-(2R,3R,5R, I'RS)-1l-t-Butoxycarbonviyl-2-(1-hydroxy-3-methyl)butvl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxvlic Acid t-Butyl Ester. A solution of (±)-(2R,3R,5R)-1l-t-butoxycarbonyl-2-formyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (7.1 g, 16.03 mmol) in diethyl ether (75 mL) was reacted with isobutyl magnesium chloride (24 mL, 2.0 M in ether, 48 mmol) at 00C for 2.5 hours. The reaction was quenched with saturated ammonium chloride and diluted with ethyl acetate. The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was used in next step without further purification. MS: (M+H) = 502 TBDMSO-, O OtBu H Boc 20F. (±)-(2R,3R,5R) 1-t-Butoxycarbonyl-2-(1-oxo-3-methyl)butyl-3-(t butyildimethylsilyloxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. A solution of oxalyl chloride (16 mL, 2M in CH 2
CI
2 ) in 100 mL of anhydrous dichloromethane was prepared and maintained under a nitrogen atmosphere, at -78 oC. DMSO (4.26 mL, 64.1 mmol) was added slowly to the solution. The mixture was stirred for 15 minutes and reacted with (±)-(2R,3R,5R,1'RS)-1-t butoxycarbonyl-2-(1-hydroxy-3-methyl)butyl-3-(t-butyldimethylsilyloxymethyl) -150- WO 99/54299 PCT/US99/07945 pyrrolidine-5-carboxylic acid t-butyl in 30 mL of anhydrous dichloromethane. The solution was stirred for 1 hour and triethylamine (17 mL, 128 mmol) was added slowly to the reaction mixture. The solution was allowed to warm slowly to room temperature, quenched with saturated sodium bicarbonate and diluted with dichloromethane. The organic layer was washed with water and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 5-10% ethyl acetate/hexanes to provide the title compound (yield: 6.3 g, 78.8 %). 1 H NMR (CDCI 3 ): 8 0.07 (m, 6H), 0.81 - 0.96 (m, 15H), 1.40 and 1.42 (two s, 9H), 1.46 and 1.47 (two s, 9H), 1.72-1.82 (m, 1H), 2.15-2.45 (m, 4H), 3.47 3.69 (m, 1H), 4.28 - 4.46 (m, 2H). MS: (M+H) = 500 TBDMSO-,
H
2 N OtBu N O HBoc 20G. (±)-(2R,3R,5R,1'RS)-1l-t-Butoxycarbonyl-2-( 1-amino-3-methy)butyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1F, substituting (±)-(2R,3R,5R) 1-t-butoxycarbonyl-2-(1-oxo-3 methyl)butyl-3-(t-butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R)-1-benzyl-2-(1-oxo-3-ethyl)pentyl-3-(t-butyl dimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.54 g, 34.1%). MS: (M+H)+= 501. -151- WO 99/54299 PCT/US99/07945 TBDMSO-, AcHN OtBu H Boc O 20H. (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-3-methy)butyl-3-(t butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1G, substituting (±)-(2R,3R,5R, 1'RS)-1 -t-butoxycarbonyl-2-(1-amino-3 methy)butyl-3-(t-butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'R)- and (±)-(2R,3R,5R,1'S)-1-benzyl-2-(1 amino-3-ethyl)pentyl-3-(t-butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 462 mg, 79.0 %). (±)-(2R,3R,5R,1'S) 'H NMR (CDCI 3 ): 5 0.03 and 0.04 (two s, 6H), 0.86 (s, 9H), 0.89 and 0.95 (two d, 6H), 1.04 (m, 1H), 1.17 -1.25 (m, 2H), 1.44 (s, 9H), 1.46 (s, 9H), 1.86 (m, 1H), 1.99 (s, 3H), 2.07 (m, 1H), 2.30 (m, 1H), 3.48 (m, 1H), 3.61 (m, 1H), 3.67 (m, 1H), 4.16 (m, 1H), 4.27 (m, 1H), 7.35 (brd, 1H). MS: (M+H) = 543.
HO
AcHNK OtBu 0 201. (±)-(2R,3R,5R,1'S)- 1-t-Butoxycarbonyl-2-(1-acetamido-3-methy)butyl-3 (hydroxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1H, substituting (±)-(2R,3R,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido 3-methy)butyl-3-(t-butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t -152- WO 99/54299 PCT/US99/07945 butyl ester in place (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(l1-acetamido-3-ethyl)butyl-3-t butyldimethylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester. MS: (M+H)*= 429. H AcHN, OtBu HN' 20J. (±)-(2R,3R,5R,1 'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-3-methy)butyl-3 formyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 2A, substituting (±)-(2R,3R,5R, 1 'S)-1l-t-butoxycarbonyl-2-(1-acetamido 3-methy)butyl-3-hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-hydroxymethyl pyrrolidine-5-carboxylic acid t-butyl ester (yield: 1.5 g, 91 %). 1 H NMR (CDCI 3 ): 5 0.92 and 0.94 (two d, 6H), 1.07 (m, 1H), 1.23-1.33 (m, 2H), 1.43 (s, 9H), 1.44 (s, 9H), 1.64 (m, 1H), 2.03 (s, 3H), 2.39 (m, 1H), 2.46 (m, 1H), 3.18 (m, 1H), 4.19 (m, 1H), 4.32 (m, 1H), 4.39 (m, 1H), 7.12 (br d, 1H). MS: (M+H) = 427 -153- WO 99/54299 PCT/US99/07945 AcHN, OtBu 20K. (±)-(2R,3S,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-3-methy)butyl-3 vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. To a suspension of methyl triphenylphosphonium bromide (125.6 mg, 0.35 mmol) in 3 ml of anhydrous toluene was added potassium t-butoxide (1.0 M in THF, 0.31 mmol) dropwise at room temperature. After stirring for 16 hours, (+) (2R,3R,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-formyl pyrrolidine-5-carboxylic acid t-butyl ester (30 mg, 0.070 mmol) in 3 ml of toluene was added dropwise and stirred for 0.5 hour. The reaction was quenched with saturated ammonium chloride and diluted with methylene chloride. The organic layer was washed with water and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using ethyl acetate to provide the title compound, as an white foamy solid (yield: 23.7 mg, 79.4%). 1 H NMR (CDCl 3 ): 6 0.92 (m, 6H), 1.26 (m, 2H), 1.44 (s, 9H), 1.47 (s, 9H), 1.65 (m, 1H), 1.97 (s, 3H), 2.43 (m, 2H), 3.56 (m, 1H), 4.15 (m, 2H), 4.32 (m, 1H), 5.11 (m, 1H), 5.15 (m, 1H), 5.75 (m, 1H), 7.35 (br, 1H). MS: (M+H) = 425. -154- WO 99/54299 PCT/US99/07945 AcHN' OH ' YN H 0 O HCI 20L. (±)-(2R,3S,5R,1'S)-2-(l1-Acetamido-3-methyl)butyl-3-vinyl-pyrrolidine-5 carboxylic Acid Hydrochloride. The title compound was prepared according to the method described in Example 1 K, substituting (±)-(2R,3S,5R, 1'S)-1 -t-butoxycarbonyl-2-(1-acetamido 3-methy)butyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R, l'S)-2-(1-acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 16.0 mg, 99.1%). 1 H NMR (DMSO-d 6 ):5 0.82 (d, 3H), 0.88 (d, 3H), 1.29 (m, 1H), 1.42 (m, 1H), 1.57 (m, 1H), 1.87 (s, 3H), 1.91 (m, 1H), 2.40 (m, 1H), 2.90 (m, 1H), 4.20 (m, 1H), 4.32 (m, 1H), 5.08 (dd, 1H), 5.17 (dd, 1H), 5.72 (ddd, 1H), 8.09 (d, 1H), 9.16 (br s, 1H), 9.28 (br s, 1H). MS: (M+H) = 269. -155- WO 99/54299 PCT/US99/07945 Example 21 (±)-(2R,3R,5R.1'S)-2-( 1 -Acetamido-3-ethyl)pentyl-3-hydroxvmethyl-pyrrolidine-5 carboxylic Acid Hydrochloride. HO-, AcHN;,, H OtBu N HH O 21A. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-hydroxymethyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1J, substituting (±)-(2R,3R,5R, 1 'S)-1 -benzyl-2-(1-acetamido-3 ethyl)pentyl-3-hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1 'S)-1 -benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. MS: (M+H) = 471 HO-, AcHN,,, OH N HH HCI 21B. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-hydroxymethyl pyrrolidine-5-carboxylic Acid Hydrochloride. The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3 hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) -156- WO 99/54299 PCT/US99/07945 (2R,3R,5R, 1l'S)-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine-5 carboxylic acid t-butyl ester 1 H NMR (DMSO-d 6 ): 6 8.15 (d, J=9Hz, 1H), 4.28-4.15 (m, 2H), 3.95-3.45 (m, 4H), 2.35 (m, 1H), 1.98 (m, 1H), 1.89 (s, 3H), 1.50-1.45 (m, 7H), 0.81 (t, J=7.4Hz, 3H), 0.77 (t, J=7.5Hz, 3H). MS: (M+H) = 301, (M-H)-= 299 Example 22 (±)-(2R,3R,5R, 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(pyrrol-1 -yl)-pyrrolidine-5 carboxylic Acid Hydrochloride. TMS O HN, AcHN,,, OtBu -..pO Ph 22A. (±)-(2S, 3R,5R, 1 'S)-1 -Benzyl-2-(1-Acetamido-3-ethyl)pentl-3-(2 trimethylsilylethoxycarbonylamino)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-carboxyl pyrrolidine-5-carboxylic acid t-butyl ester (80 mg, 0.18 mmol) prepared according to the procedure of Example 2B was reacted with diphenylphosphoryl azide (0.047 mL, 0.216 mmol), 2-trimethylsilylethanol (0.034 mL, 0.234 mmol), and triethylamine (0.030 mL, 0.216 mmol) in toluene (2 mL) at 750C for 15 hours. The reaction was concentrated in vacuo and the resulting residue purified by chromatography on silica gel using 25% ethyl acetate/hexanes to provide the title compound, as a light yellow oil (yield: 46 mg, 45%). MS: (M+H) = 576, (M+Na) = 598, (M-H) = 574. -157- WO 99/54299 PCT/US99/07945
H
2 N. AcHN/,, OtBu Ph 22B. (±)-(2S,3R,5R,1 'S)-l1-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-amino pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 1H, substituting (±)-(2S,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-(2-trimethylsilylethoxycarbonylamino)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-(t butyldimethylsilyloxmethyl)-pyrrolidine-5-carboxylic acid t-butyl ester. MS: (M+H)*= 432, (M-H)- = 430. AcHN,, H OtBu N Ph 22C. (+)-(2S,3R,5R, 1 'S)-1l-Benzyl-2-(1l-Acetamido-3-ethyl)pentyl-3-(pyrrol-1-yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2S,3R,5R, 1'S)-1 -Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-amino pyrrolidine-5-carboxylic acid t-butyl ester (34 mg, 0.078 mmol) was reacted with 40% succinic dialdehyde in water (50 mg, 0.234 mmol), acetic acid (0.00044 mL, 0.0078 mmol), and 4A molecular sieves (200 mg) in toluene (2 mL) at RT for 3 hours. The reaction was concentrated in vacuo and the resulting residue purified -158- WO 99/54299 PCT/US99/07945 by chromatography on silica gel using 50% ethyl acetate/hexanes to provide the title compound, as an oil (yield: 7.1 mg, 19%). MS: (M+H) = 482, (M+Na) + = 504, (M-H)- = 480. "/ AcHN,, . OtBu . N 'i H 22D. (±)-(2S,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(pyrrol-1-yl) pyrrolidine-5-carboxylic Acid t-Butyl ester. The title compound is prepared according to the method described in Example 1 J, substituting (±)-(2S,3R,5R,1'S)-1l-benzyl-2-(1 -acetamido-3 ethyl)pentyl-3-(pyrrol- 1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1'S)-l1-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester (yield: 3.5 mg, 61%). MS: (M+H) = 392, (M-H) = 390. -159- WO 99/54299 PCT/US99/07945 \ N, /. AcHN,, . OH HOCI 22E. (±)-(2S.3R,5R,1'S)-2-(1-Acetamido-3-ethyl)penyl-3-(pyrrol-1-yl) pyrrolidine-5-carboxylic Acid Hydrochloride. The title compound was prepared according to the method described in Example 1K, substituting (±)-(2S,3R,5R, l'S)-2-(1-acetamido-3-ethyl)pentyl-3 (pyrrol-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 3.5 mg, 100%). 1 H NMR (D20) 5 7.48 (bs, 1H), 6.77 (bs, 2H), 5.97 (bs, 2H), 4.33 (m, 1H), 3.70 (m, 1H), 3.07 (m, 1H), 2.43 (m, 1H), 1.92 (m, 1H), 1.75 (s, 3H), 1.55 (m, 1H), 1.35-1.10 (m, 7H), 0.81 (m, 3H), 0.75 (m, 3H). MS: (M+H)* = 336, (M-H) = 334. -160- WO 99/54299 PCT/US99/07945 Example 23 (±)-(2R, 3R, 5R, 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(1 -cis-N-hydroxyimino)ethyl pyrrolidine-5-carboxylic Acid Hydrochloride. NOH
H
3 C t AcHN,, , OtBu Ph 23A. (±)-(2R,3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(1-cis-N hydroxyimino)ethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R, 1'S) 1-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-acetyl pyrrolidine-5-carboxylic acid t-butyl ester (45 mg, 0.1 mmol) prepared according to the method of Example 8B in methanol/methylene chloride (3/1) was reacted with a solution of hydroxylamine hydrochloride (21 mg, 0.3 mmol) and sodium hydroxide (12 mg, 0.3 mmol) in methanol (2 mL) for 2h. The reaction was diluted with ethyl acetate. The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 40% ethyl acetate/hexanes to provide the cis oxime title compound (lower Rf spot on TLC), as an oil (yield: 35 mg, 75%), as well as the trans-oxime title compound (higher Rf spot on TLC), as an oil (yield: 13 mg, 25%). MS: (M+H) = 474 -161- WO 99/54299 PCT/US99/07945 NOH H3C . AcHN,, ~ OtBu N 'If H 0 /O 23B (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3(1-cis-N hydroxyimino)ethyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 1J, substituting (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-(1-cis-N-hydroxyimino)ethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 methoxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (CDCl 3 ): 8 3.92 (br, 1H), 3.70 (m, 2H), 2.82 (m, 1H), 2.38 (m, 1H), 1.88 (s, 3H), 1.78 (s, 3H), 1.39 (s, 9H), 1.40-1.20 (m, 7H), 0.76 (m, 6H). MS: (M+H) = 384 NOH
H
3 C-4 AcHN,, OH N '"if HC 23C. (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-(1-cis-N hydroxyimino)ethyl-pyrrolidine-5-carboxylic Acid Hydrochloride The title compound is prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R,1'S)-2-(1 -acetamido-3-ethyl)pentyl-3-(1 cis-N-hydroxyimino)ethyl-5-carboxylic acid t-butyl ester in place of (+) -162- WO 99/54299 PCT/US99/07945 (2R,3R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5 carboxylic acid t-butyl ester. 1 H NMR (D 2 0): 68 4.35 (m, 1H), 4.00 (m, 1H), 3.80 (m, 1H), 3.71 (m, 1H), 3.63 (m, 1H), 3.13 (q, J=8.4Hz,, 1H), 2.64 (m, 1H), 2.18 (m, 1H), 1.97 (s, 3H), 1.85 (s, 3H), 1.50-1.10 (m, 7H), 0.77 (m, 6H). MS: (M+H) = 328 Example 24 (±)-(2R, 3R, 5R, 1 'S)-2-(1 -Acetamido-3-ethyl)pentyl-3-(N-hydroxyimino)methyl pyrrolidine-5-carboxylic Acid Hydrochloride. HNOH AcHN .. OtBu N H 0 24A. (±)-(2R,3R,5R,1 'S)-2-(1-Acetamido-3-ethyl)pentvl-3-(N hydroxyimino)methyl-pyrrolidine-5-carboxvlic Acid t-Butyl Ester. (±)-(2R,3R,5R, l'S)-1l-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-formyl pyrrolidine-5-carboxylic acid t-butyl ester (18 mg, 0.051 mmol) prepared according to the method of Example 2A was reacted with hydroxylamine hydrochloride (7 mg, 0.11 mmol) in 1N NaOH in methanol (3 mL) at 250C for 1.5 hours. The reaction was quenched with aqueous ammonium chlororide (3ml) and water (3ml)and taken by dichloromethane (2x 10 ml). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 5% methanol in dichloromethane to provide the title compound, as an oil (yield: 6 mg, 32%). -163- WO 99/54299 PCTIUS99/07945 MS: (M+H)+=370 NOH H AcHN.. OH N H 0 HCI 24B (±)-(2R,3R,5R,1 'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(N hydroxyimino)methyl-pyrrolidine-5-carboxylic Acid Hydrochloride The title compound is prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R, 1 'S)-2-(1-acetamido-3-ethyl)pentyl-3-(N hydroxyimino)methyl-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5 carboxylic acid t-butyl ester. Example 25 (±)-(2R,3R,5R. 1 'S)-2-(1-Acetamido-3-ethyl)pentyl-3-(methoxyimino)methyl pyrrolidine-5-carboxylic Acid.
NH
2
CH
3 0 AcHN.. OH N K Ph 25A. (±)-(2R,3R,5R,1'S) 1-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3 (methoxyimino)methyl-pyrrolidine-5-carboxylic Acid (±)-(2R,3R,5R,1'S) 1-Benzyl-2-(1-acetamido-3-ethyl)pentyl-3-cyano pyrrolidine-5-carboxylic acid t-butyl ester (20 mg, 0.045 mmol) prepared according to the method of Example 17E was reacted with hydrogen chloride -164- WO 99/54299 PCT/US99/07945 (0.45 mmol) in ether (2 mL) and methanol (0.1 mL) at 0 0 C for 5 hours. The reaction was neutralized with aqueous ammonium hydroxide and purified on silica gel with 3% methanol in dichloromethane to provide the title compound, as a white solid (yield: 5 mg, 26%). MS: (M+H)*=418 NH CH30 AcHN. OH N H 0 25B. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-ethyl)pentyl-3 (methoxyimino)methyl-pyrrolidine-5-carboxylic Acid. The title compound is prepared according to the method described in Example 1J, substituting (±)-(2R,3R,5R,1'S) 1-Benzyl-2-(1-acetamido-3 ethyl)pentyl-3-(imino-methoxymethyl)-pyrrolid ine-5-carboxylic acid t-butyl ester, in place of (±)-(2R,3R,5R,1'S)-1-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 methoxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 3.9 mg, 96%). 1 H NMR (DMSO-d 6 ) 8 7.52(d, J=8.7 HZ, 1H), 7.15(s, 1H),6.77(s, 1H), 3.68(m, 1H),3.61(s, 3H), 3.22(m, 1H), 2.51(m, 1H), 2.23(m, 1H), 1.82(m, 1H), 1.78(s, 3H), 1.40(m, 1H), 1.26(m, 3H), 1.13(m, 3H), 0.78(t, J=6.5HZ, 3H), 0.72(t, J=6.5HZ, 3H) MS: (M+H)*=328 -165- WO 99/54299 PCTIUS99/07945 Example 26 (+)-(2R,3R,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(hydroxyacetyl)-pyrrolidine 5-carboxylic Acid Hydrochloride. HO OH AcHN, OtBu N H Boc o 26A. (±)-(2R,3R.,5R,1'S,1"RS)-1l-t-Butoxycarbonyl-2-(1-acetamido-3 methy)butyl-3-(1,2-dihydroxy)ethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 20A substituting (±)-(2R,3R,5R, 1 'S)-1l-t-butoxycarbonyl-2-(1 -acetamido 3-methy)butyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester for (±)-(2S,3R,5R) 1-benzyl-2-vinyl-3-(t-butyldimethylsilyloxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester. HO 0O AcHN, KOtBu N H Boc O 26B. (±)-(2R,3R.5R,1'S)-l-t-Butoxycarbonyl-2-(1-acetamido-3-methy)butyl-3 (hydroxyacetyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'S,l1"RS)-1l-t-Butoxycarbonyl-2-(1-acetamido-3 methy)butyl-3-(1,2-dihydroxy)ethyl-pyrrolidine-5-carboxylic acid t-butyl ester is reacted with dibutyltin oxide in methanol according to the procedure of Kong in J. Carbohydrate Chem. 1993, p. 557. The reaction is concentrated and the residue -166- WO 99/54299 PCT/US99/07945 is redissolved in dichloromethane and reacted with bromine as described in the above reference to give the title compound. HO 0 AcHN OH 'N ]< H HCI 260. (±)-(2R. 3R,5R,1'S)-2-(1-Acetamido-3-methy)butyl-3-(hydroxyacetyl) pyrrolidine-5-carboxylic Acid Hydrochloride. The title compound is prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R, 1 'S)-1l-t-butoxycarbonyl-2-(1-acetamido 3-methy)butyl-3-hydroxyacetyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1 'S)-2-(1-acetamido-3-ethyl)pentyl-3-(methoxymethyl) pyrrolidine-5-carboxylic acid t-butyl ester. -167- WO 99/54299 PCT/US99/07945 Example 27 (±)-(2S,3R, 5R ,1'S)-2-(1 -Acetamido-3-methyl)butyl-3-amino-pyrrolidine-5 carboxylic Acid Dihydrochloride. CbzHN,/ HO OtBu S Ph 27A. (±)-(2S,3R,5R,1'RS)-1l-Benzyl-2-(1-hydroxy-3-methyl)butyl-3 benzyloxycarbonylamino-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 9D, substituting isobutylmagnesium bromide in place of 3 pentylmagnesium bromide. CbzHN,/. AcHN .OBu HN "'I - L.o Ph 27B. (±)-(2S,3R,5R,1'S)-1l-Benzyl-2-(1l-acetamido-3-methyl)butyl-3 benzyloxycarbonylamino-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Examples 9E-H substituting (±)-(2R,3R,5R,1'RS)-1l-benzyl-2-(l1-hydroxy-3 methyl)butyl-3-benzyloxycarbonylamino-pyrrolidine-5-carboxylic acid t-butyl ester for (±)-(2R,3R,5R, 1 'RS)-1l-benzyl-2-(1-hydroxy-3-ethyl)pentyl-3 benzyloxycarbonylamino-pyrrolidine-5-carboxylic acid t-butyl ester as the starting material of the sequence in Example 9E. -168- WO 99/54299 PCT/US99/07945
H
2 N,. AcHN,, OH HN H0 2 HCI 27C. (±)-(2S,3R,5R, 1'S)-2-(1-acetamido-3-methyl)butyl-3-amino-pyrrolidine-5 carboxylic Acid Dihydrochloride The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-methyl)butyl-3 amino-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2 (1-acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5-carboxylic acid t butyl ester. 1 H NMR (d 6 -DMSO) 8 8.64( bs, 1H), 8.32 (bs, 1H), 8.23 (bs, 1H), 8.18 (d, J=6Hz, 1 H), 4.79 (d, J=7Hz, 1H), 4.42 (m, 1 H), 4.33 (m, 1H), 4.21 (m, I1H), 4.07 (m, IH), 3.76 (m, 2H), 2.73 (m, 2H), 1.92 (m, 3H), 0.80-0.97 (m, 7H). MS (M+H)* = 258 -169- WO 99/54299 PCT/US99/07945 Example 28 (±)-(2R,3R, 5R,1'S)-2-(1 -Acetamido-3-methyl)butyl-3-methoxycarbonyl pyrrolidine-5-carboxylic Acid Hydrochloride. HO O,// AcHN" OtBu H BocO 28A. (±)-(2R,3R,5R.,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 carboxyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 2B, substituting (±)-(2R,3R,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R, 1'S)-1 -benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-formyl-pyrrolidine-5 carboxylic acid t-butyl ester.
CH
3 0 AcHN, OtBu H Boc 28B. (±)-(2R,3R.5R, 1'S)-1-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 methoxycarbonyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 2C, substituting (±)-(2R,3R,5R, 1 'S)-1l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-carboxyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-carboxyl-pyrrolidine-5 carboxylic acid t-butyl ester. -170- WO 99/54299 PCT/US99/07945 0
CH
3 0 AcHN , "KOH N H0 / O HCI 28C. (±)-(2R,3R,5R,1'S)-2-(1-Acetamido-3-methyl)butyl-3-methoxycarbonyl pyrrolidine-5-carboxylic Acid Hydrochloride. The title compound was prepared according to the method described in Example 2E, substituting (±)-(2R,3R,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-methoxycarbonyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-methoxycarbonyl pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (DMSO-d 6 ): 8 8.24, 8.08 (d, J=9Hz, 1H), 4.44, 4.36 (m, 1H), 4.25, 4.15 (m, 1H), 3.98, 3.88 (m, 1H), 3.65, 3.64 (s, 3H), 3.18, 3.10 (m, 1H), 2.57, 2.20 (m, 2H), 1.87, 1.83 (s, 3H), 1.57 (m, 2H), 1.36 (m, 1H), 0.88 (d, J=7.5Hz, 3H), 0.82 (d, J=7.5Hz, 3H). MS: (M+H) = 301 -171- WO 99/54299 PCT/US99/07945 Example 29 (±)-(2R, 3R,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(imidazol-2-yl)-pyrrolidine-5 Carboxylic Acid Dihydrochloride. NH AcHN,, O-t-Bu o Ph 29A. (±)-(2R,3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-methyl)butvli-3-(imidazol-2 yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 15A, substituting (±)-(2R,3R,5R, 1'S)-1l-benzyl-2-(1-acetamido-3 methyl)butyl-3-forrnyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R, 1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-formyl-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 27.4 mg, 83. %). MS: (M+H) = 455. rNH AcHN,, O-t-Bu N H 0 29B. (±)-(2R,3R,5R, 1'S)-2-(1-Acetamido-3-methyl)butyl-3-(imidazol-2-yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 15B, substituting (±)-(2R,3R,5R, 1 'S)-1l-benzyl-2-(1 -acetamido-3 methyl)butyl-3-(imidazol-2-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of -172- WO 99/54299 PCT/US99/07945 (±)-(2R,3R,5R, 1'S)-1 -benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(imidazol-2-yl) pyrrolidine-5-carboxylic acid t-butyl ester (yield: 19.1 mg, 95.5%). MS: (M+H) = 365. NH N --. AcHN/,, * OH N HH 2 HCI 29C (±)-(2R,3R,5R,1 'S)-2-(1-Acetamido-3-methyl)butyl-3-(imidazol-2-yl) pyrrolidine-5-Carboxylic Acid Dihydrochloride. The title compound was prepared according to the method described in Example 15B, substituting (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3 methyl)butyl-3-(imidazol-2-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-(imidazol-2-yl) pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (DMSO-d6): 8 0.76 (d, J = 6.6 Hz, 3H), 0.82 (d, J = 6.6 Hz, 3H), 1.18 (t, 2H), 1.44 (m, 1H), 1.71 (s, 3H), 2.43-2.47 (m, 1H), 2.80 (m, 1H), 3.83 (m, 1H), 4.05 (m, 1H), 4.28 (m, 1H), 4.55 (t, 1H), 7.65 (s, 2H), 8.03 (d, J = 8.4 Hz, 1H). MS: (M+H)*= 326. -173- WO 99/54299 PCT/US99/07945 Example 30 (±)-(2R.,3R,5R,1'S)-2-(1-Acetamido-3-methyl)butl-3-(imidazol-4-l)-pyrrolidine-5 carboxylic Acid Dihydrochloride. 30A. (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-3-methy)butyl-3 carboxyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. 0 O HO-j1.. AcHN,, OtBu N "rIf .0 PH The title compound was prepared according to the method described in Example 2B, substituting (±)-(2R,3R,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido 3-methy)butyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester prepared according to the method described in Example 20J in place (±)-(2R,3R,5R,1'S)-1 benzyl-2-(1-acetamido-3-ethyl)pentyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 129.5 mg, >100 %). MS: (M+H)*= 443. O N2 0 AcHN,, OtF3u 0 PH 30B. (±)-(2R,3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-methyl)butyl-3-diazoaceyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 12A, substituting (±)-(2R,3R,5R,1'S)-1 -benzyl-2-(1-acetamido-3 -174- WO 99/54299 PCT/US99/07945 methyl)butyl-3-carboxyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R,1'S)-1-benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-carboxyl-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 218.8 mg, 100%). MS: (M+H) = 458. O Br 0 AcHN,, OtBu 0 PH 30C. (±)-(2R,3R,5R,1 'S)-1l-Benzyl-2-(1-acetamido-3-methyl)butyl-3 bromoacetyl-pyrrolidine-5-carboxylic Acid t-Butlyl Ester. The title compound was prepared according to the method described in Example 12B, substituting (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3 methyl)butyl-3-diazoacetyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-diazoacetyl pyrrolidine-5-carboxylic acid t-butyl ester (yield: 107.2 mg, 45.5 %). 1 H NMR (CDCi 3 ): 8 0.90 (d, 6H), 1.26 - 1.35 (m, 3H), 1.42 (s, 9H), 1.95 (s, 3H), 2.25 (m, 2H), 3.11 (m, 1H), 3.54 (dd, 1H), 3.69 (m, 1H), 3.93 (dd, 2H), 4.11 (d, 1H), 4.27 (m, 1H), 4.35 (d, 1H), 5.05 (br d, 1H), 7.25-7.32 (m, 5H). MS: (M+H) = 509. -175- WO 99/54299 PCT/US99/07945 /"N AcHN,, OtBu HN Ph 30D. (±)-(2R.3R,5R,1'S)-1l-Benzyl-2-(1-acetamido-3-methyl)butyl-3-(imidazol-4 yl)-pyDvrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 12C, substituting (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3 methyl)butyl-3-bromoacetyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1'S)-1 -benzyl-2-(1-acetamido-3-ethyl)pentyl-3-bromoacetyl pyrrolidine-5-carboxylic acid t-butyl ester (yield: 32.3 mg, 60.4%). MS: (M+H)* = 455. H N HN\) AcHN, OtBu ~O H 0 30E. (±)-(2R,3R.5R,1'S)-2-(1-Acetamido-3-methyl)butyl-3-(imidazol-4-yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1 J, substituting (±)-(2R,3R,5R, 1 'S)-1 -benzyl-2-(1-acetamido-3 methyl)butyl-3-(imidazol-4-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1 'S)-1 -benzyl-2-(1 -acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester (yield: 23.9 mg, 96.2%). -176- WO 99/54299 PCT/US99/07945 1 H NMR (CDCI 3 ): 8 0.87 (d, 3H), 0.89 (d, 3H), 1.26 (m, 1H), 1.41 (m, 2H), 1.46 (s, 9H), 1.59 (m, 1H), 1.93 (s, 3H), 2.62 (m, 1H), 3.30 (m, 1H), 3.54 (m, 1H), 3.79 (m, 1H), 4.01 (m, 1H), 6.11 (brd, 1H), 6.89 (s, 1H), 7.63 (s, 1H). MS: (M+H) = 365. HNN AcHN,, . OH N "If, HH 2 HCI 30F. (±)-(2R,3R,5R,1 'S)-2-(l1-Acetamido-3-methyl)butyl-3-(imidazol-4-yl) pyrrolidine-5-carboxylic Acid Dihydrochloride. The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R,1'S)-2-(1l-acetamido-3-methyl)butyl-3 (imidazol-4-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5 carboxylic acid t-butyl ester to provide the title compound solid (yield: 24.4mg, 100%). 1 H NMR (DMSO-d 6 ): 8 0.76 (d, J = 3.6 Hz, 3H), 0.88 (d, J = 3.6 Hz, 3H), 1.22 (m, 1H), 1.28 (m, 1H), 1.48 (m, 1H), 1.79 (s, 3H), 2.32 (dt, 1H), 2.71 (dt, 1H), 3.68 (m, 1H), 3.96 (m, 1 H), 4.28 (m, 1 H), 4.51 (t, 1H), 7.63 (s, 1H), 8.23 (d, J = 5.1 Hz, 1H), 9.10 (s, 1H), 9.67 (br s, 1 H), 14.51 (brs, 1H). MS: (M+H)*= 309. -177- WO 99/54299 PCT/US99/07945 Example 31 (±)-(2R,3R,5R, 1 'S)-2-( 1 -Acetamido-3-methvyl)butyl-3-(thiazol-4-yl)-pyrrolidine-5 carboxylic Acid Dihydrochloride. / N AcH N ,, ,OtBu HBoc o 31A. (±)-(2R, 3R, 5R, I'S)-1 -t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 (thiazol-4-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 bromoacetyl-pyrrolidine-5-carboxylic acid t-butyl ester (36.5 mg, 0.07 mmol) was reacted with thioformamide (21.4 mg, 0.35 mmol) in ethanol (5 ml) at reflux for 4 hours. The reaction was concentrated in vacuo. The residue was treated with 5 ml of aqueous NaHCO 3 and extracted with dichloromethane (4 x 5 ml). The organic layers were washed with brine, dried over Na 2
SO
4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using ethyl acetate to provide the title compound, as a white solid (yield: 23.8 mg, 70.4%). MS: (M+H)Y= 482. -178- WO 99/54299 PCT/US99/07945 / N S AcHN/, OH N '1 H 0 2 HCI 31B. (±)-(2R,3R,5R,1'S)-2-(l1-Acetamido-3-methvl)bulyl-3-(thiazol-4-yl) pyrrolidine-5-carboxylic Acid Dihydrochloride. The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R, 1 'S)-l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-(thiazol-4-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2-(1 -acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 18.5 mg, 100%). 1 H NMR (DMSO-d 6 ): 5 0.62 (d, J = 4.2 Hz, 3H), 0.72 (d, J = 4.2 Hz, 3H), 1.05 (m, 1H), 1.12 (m, 1H), 1.30 (m, 1H), 1.72 (s, 3H), 2.14 (dt, 1H), 2.59 (dt, 1H), 3.69 (m, 1H), 3.92 (br m, 1H), 4.21 (m, 1H), 4.38 (br m, 1H), 7.46 (d, J = 1.2 Hz, 1H), 8.02 (d, J = 5.1 Hz, 1H), 9.04 (d, J = 1.2 Hz, 1H), 9.39 (br s, 1H), 9.48 (br s, 1H). .MS: (M+H)*= 326. -179- WO 99/54299 PCT/US99/07945 Example 32 (±)-(2R,3R. 5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(thiazol-2-yl)-pyrrolidine-5 carboxylic Acid Dihydrochloride. O
H
2
N
' AcHN.. OtBu ' N H Boc 32A. (±)-(2R.3R,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 carbamoyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R, 1'S)-1 -t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 carboxyl-pyrrolidine-5-carboxylic acid t-butyl ester (0.258g , 0.584 mmol) was reacted with isobutyl chloroformate (80 mg, 0.84 mmol) and N-methylmorpholine (59 mg, 0.584 mmol) in THF (10 mL) at 0 0 C for 0.25 hours. Aqueous ammonium hydroxide (.39 mL) was added and the reaction was stirred at 0 0 C for 0.5 hours. The reaction was diluted with ethyl acetate. The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 100% ethyl acetate to 5 % methanol-ethyl acetate to provide the title compound, as a glass (yield: 182 mg, 70.7 %). 1 H NMR (CD 3 0D) 8 4.70(m,1H), 4.36 (q, J=3 Hz,1H), 4.05 (m,1H),2.87( q oft, J=9 and 3 Hz, 1H), 2.52 (m, 1H), 2.36 (m, 1H) 1.94 (d, 3H),1.63 (m, 1H), 1.41-1.53 (m, 18H), 1.3 (m, 2H), 0.9-0.18 (m, 6H) MS: (M+H) = 442 -180- WO 99/54299 PCT/US99/07945 S
H
2 N' AcHN. OtBu H Boc O 32B. (±)-(2R,3R,5R,1'S)- 1-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 thiocarbamoyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-3-methy)butyl-3 carbamoyl-pyrrolidine-5-carboxylic acid t-butyl ester (70 mg, 0.159 mmol) was reacted with P 2
S
1 0 (8.5 mg, 0.019 mmol) in 4 ml tetrahydrofuran and 1 ml of methylene chloride at room temperature. After 1.25 hrs, 9.6 mg of P 2
S
1 0 was added. The starting material had been consumed after 2 hrs. The mixture was diluted with ethyl acetate, washed with water and brine, dried over MgsO 4 , filtered and concentrated. Tic analysis showed two spots and the mass spectrum indicated it was a mixture of mono-thio and di-thio compounds. The material was used in the next reaction without further purification. MS: (M+H) = 458, 474 AcHN,, tBu 'N H Boc O 32C. (±)-(2R,3R,5R,1'S)-l-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 (thiazol-2-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1 'S)- l-t-Butoxycarbonyl-2-(1-acetamido-3-methy)butyl-3 thiocarbamoyl-pyrrolidine-5-carboxylic acid t-butyl ester (73 mg, o.16 mmol) was reacted with chloroacetaldehyde (50% in water) (0.02 ml, 0.16 mmol) in 5 ml of -181- WO 99/54299 PCT/US99/07945 acetone at 750C. Magnesium sulfate (0.9 g) and additional chloroacetaldehyde was added at intervals over the next 5 hr when till complete conversion of starting material. The reaction was diluted with ethyl acetate, washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 100% ethyl acetate to provide the title compound, as a glass (yield: 12.6 mg, 16.3%). 1 H NMR (CDCl 3 ) 5 7.69(m, 1H), 7.45(m, 1H), 4.44 (m, 1H), 4.28 (m, 2H), 3.52(m, !H), 2.7 (m, 1H), 2.5 (m, 1H), 1.99 (s, 3H), 1.44 (s, 9H), 1.37 (s, 9H), 1.27 (m, 3 H), 0.95 (m, 6 H). MS: (M+H) = 482 S. AcHN/,, . OH N H 0 2 HCI 32D. (±)-(2R,3R,5R,1'S)-1- t-Butoxycarbonyl-2-(1l-acetamido-3-methyl)butyl-3 (thiazol-2-yl)-pyrrolidine-5-carboxylic Acid Dihydrochloride The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R, 1 'S)-1l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-(thiazol-2-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 10.1 mg, 82%). 1 H NMR (DMSO-d 6 ) 8 8.1 (d, J=10OHz, 1H), 7.79 (d, J--4Hz, 1H), 7.69 (d-4 Hz, 1H), 4.49 (t, J=7.5, 1H), 4.22 (m, 1H), 4.14 (t, J=9Hz, 1H), 4.01 (q, J=10Hz, 1H), 2.80 (m, 1H), 2.25 (m, 1H), 1.78 (s, 3H), 1.47 (m, 1H), 1.25 (m, 2H), 0.83 (d, J=6.2Hz, 3H), 0.75 (d, J=6.2Hz, 3H) -182- WO 99/54299 PCT/US99/07945 MS: (M-H)- = 324, (2M-1)-= 649, (M+35) = 360 Example 33 (±)-(2R, 3S,5R,1'S)--2-(1 -acetamido-3-methyl)butyl-3-(cis-2-chloro-vin-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt CI Cl AcHN. Bu AcHN OtBu N NP (0 HBoc O H Boc O 33A. (±)-(2R,3S,5R,1'S)-l-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 (cis-2-chloro-vin-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester and (±) (2R,3S,5R,1'S)-l-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-(trans-2 chloro-vin-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 20K substituting (chloromethyl)triphenylphosphonium chloride in place of methyltriphenylphosphonium bromide. The higher Rf 0.73 (ethyl acetate) new spot was identified to be the cis- isomer (yield: 38.4 mg, 40 %) and the lower Rf 0.57 (ethyl acetate) spot trans- isomer (yield: 42 mg, 43 %). cis- isomer 'H NMR (CDCI 3 ): 87.44 (br, 1H), 6.13 (d, J=7.5Hz, 1H), 5.32 (dd, J=9Hz, J=7.5Hz, 1H), 4.31-4.16 (m, 2H), 3.65 (m, 1H), 3.12 (m, 1H), 2.50 (m, 1H), 1.98 (s, 3H), 1.62 (m, 1H), 1.47 (s, 9H), 1.45 (s, 9H), 1.30-1.07 (m, 2H), 0.82 (m, 6H) MS: (M+H)*= 459 -183- WO 99/54299 PCT/US99/07945 trans- isomer 1 H NMR (CDCI 3 ): 5 6.12-5.90 (m, 2H), 4.30-4.07 (m, 2H), 3.64 (m, 1H), 2.62-2.37 (m, 2H), 1.98 (s, 3H), 1.69 (m, 1H), 1.48 (s, 9H), 1.45 (s, 9H), 1.26 (m, 2H), 0.91 (m, 6H). MS: (M+H)*= 459 Cl AcHN OH N HH O TFA 33B. (±t)-(2R,3S,5R,1'S)-2-(1-acetamido-3-methyl)butyl-3-(cis-2-chloro-vin-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. (+)-(2R,3S,5R, l'S)-1 -t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 (cis-2-chloro-vin-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (10 mg, 0.022 mmol) was reacted with trifluoroacetic acid (1.8 mL) in dichloromethane (0.4 mL) at room temperature for 7 hours. The reaction was concentrated in vacuo. The residue was dried on high vacuum to provide the title compound. 'H NMR (DMSO-d 6 ): 6 8.015 (d, J=7.63Hz, 1H), 6.42 (d, J=7.02Hz, 1H), 5.89 (dd, J=7.02Hz, J=8.7Hz, 1H), 4.42 (m, 1H), 4.17 (m, 1H), 3.59 (m, 1H), 3.31 (m, 1 H), 2.47 (m, 1 H), 1.88 (s, 3H), 1.84 (m, 1 H), 1.58 (m, 1 H), 1.39 (m, 1 H), 1.29 (m, 2H), 0.885 (d, J=6.71Hz, 3H), 0.83 (d, J=6.41, 3H). MS: (M+H)* = 303 -184- WO 99/54299 PCT/US99/07945 Example 34 (±)-(2R,3S,5R,1'S)-2-(1-acetamido-3-methyl)butyl-3-(trans-2-chloro-vin-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt CI AcHN. OH N HH TFA 34B. (±)-(2R,3S,5R,1 'S)-2-(1-acetamido-3-methyl)butyl-3-(trans-2-chloro-vinyl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. The title compound was prepared according to the method described in Example 33B, substituting (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-(trans-2-chloro-vin-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1 'S)-1 -t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl 3-(cis-2-chloro-vin-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (DMSO-d 6 ): 5 8.04 (d, J=7.93Hz, 1H), 6.355 (d, J=13.1 Hz, 1H), 5.93 (dd, J=13.1 Hz, J=9.32 Hz, 1H), 4.33 (m, 1H), 4.19 (m, 1H), 2.95 (m, 1H), 2.40 (m, 1H), 1.94 (m, 1H), 1.88 (s, 3H), 1.58 (m, 1H), 1.39 (m,1H), 1.29 (m, 1H), 0.89 (d, J=6.7 Hz, 3H), 0.825 (d, J=6.7 Hz, 3H). MS: (M+H)*= 303 Example 35 (±)-(2R,3S,5R,1'S)-2-(1-acetamido-3-methl)butyl-3-(cis-propen-1-yl)-pyrrolidine 5-carboxylic Acid Trifluoroacetic Acid Salt -185- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OBu 'N H Boc 35A. (±)-(2R.,3R,5R,1'S)-l-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyil-3-( cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. To a suspension of ethyl triphenylphosphonium bromide (479 mg, 1.29 mmol) in 3 mL anhydrous toluene was added potassium t-butoxide (1.0 M in THF, 0.94 mmol) dropwise at room temperature. After stirring for 2.5 hours, (±) (2R,3R,5R,1'S)-1 -t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-formyl pyrrolidine-5-carboxylic acid t-butyl ester (90 mg, 0.211 mmol) in 5 mL toluene was added dropwise and stirred for 1 hour. The reaction was quenched with saturated aqueous ammonium chloride and diluted with ethyl acetate. The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 50% ethyl acetate/hexanes to provide the title compound, as an oil (yield: 70.6 mg, 76%). MS: (M+H) = 439
CH
3 AcHN OH Nf' H H TFA 35B. (±)-(2R,3S,5R, 'S)-2-(1-acetamido-3-methyl)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. The title compound was prepared according to the method described in Example 33B, substituting (±)-(2R,3S,5R, 1 'S)-l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in -186- WO 99/54299 PCT/US99/07945 place of (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 (cis-2-chloro-vinyl)-pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (DMSO-d 6 ): 8 8.04 (d, J=7.5Hz, 1H), 5.51 (m, 1H), 5.26 (m, 1H), 4.32 (m, 1H), 4.18 (m, 1H), 3.45 (m, 1H), 3.18 (m, 1H), 2.39 (m, 1H), 1.88 (s, 3H), 1.73 (m, 1H), 1.63 (dd, 3H), 1.58 (m, 1H), 1.38 (m, 1H), 1.28 (m, 1H), 0.88 (d, J=6Hz, 3H), 0.81 (dd, J=6Hz, 3H). MS: (M+H) = 283 Example 36 (±)-(2R,3S,5R,1'S)-2-(1 -acetamido-3-methyl)butyl-3-(2,2-dimethyl-vin-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3
CH
3 AcHN N H BocO O 36A. (±)-(2R,3S.5R, l'S)-1-t-Butoxycarbonyl-2-(1 -acetamido-3-methyl)butyl-3 (2,2-dimethyl-vin-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 20K substituting isopropyl triphenylphosphonium iodide in place of methyltriphenylphosphonium bromide (yield: 22.6 mg, 33 %). 'H NMR (CDCI 3 ): 5 7.77 (d, 1H), 5.06 (d, J=10OHz, 1H), 4.18 (m, 2H), 3.50 (m, 1H), 2.69 (m, 1H), 2.32 (m, 1H), 1.97 (s, 3H), 1.70 (s, 3H), 1.64 (s, 3H), 1.65 -187- WO 99/54299 PCT/US99/07945 (m, 1H), 1.47 (s, 9H), 1.44 (s, 9H), 1.30-1.00 (m, 3H), 0.93 (d, J=6Hz, 3H), 0.88 (d, J=6Hz, 3H). MS: (M+H)% = 453
CH
3
CH
3 AcHN. OH 'N HH O TFA 36B. (±)-(2R,3S,5R,1'S)-2-(1-acetamido-3-methyl)butvl-3-(2,2-dimethyl-vin-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. The title compound was prepared according to the method described in Example 33B, substituting (±)-(2R,3S,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-(2,2-dimethyl-vin-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1 'S)-l-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl 3-(cis-2-chloro-vin-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (DMSO-d 6 ): 5 8.01 (d, J=7.5HZ, 1H), 4.99 (d, J=10OHz, 1H), 4.30 (m, 1H), 4.14 (m, 1H), 3.40 (m, 1H), 3.06 (m, 1H), 2.36 (m, 1H), 1.86 (s, 3H), 1.66 (s, 3H), 1.63 (s, 3H), 1.57 (m, 1H), 1.39-1.20 (m, 3H), 0.88 (d, J=6Hz, 3H), 0.81 (d, J=6Hz, 3H). MS: (M+H)*= 297 Example 37 (±)-(2R,3S,5R. 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-(2,2-difluoro-vin-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt -188- WO 99/54299 PCT/US99/07945 F F AcHN OtBu 'N H Boc O 37A. (±)-(2R,3S,5R,1'S)-1-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 (2,2-difluoro-vin-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. n-Butyllithium (1.6M in hexanes, 0.61 mL, 0.97 mmol) was added to diisopropylamine (136 gL, 0.97 mmol) in 4 mL THF at -78 OC and stirred for 30 min. diethyl difluoromethylphosphonate (182 mg, 0.97 mmol) was added, the colorless solution changed slowly to yellow after stirring at -78 °C for 2 hours. (±)-(2R,3R,5R, 1'S)-1 -t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-formyl pyrrolidine-5-carboxylic acid t-butyl ester (59 mg, 0.138 mmol) in 3 mL THF was added, stirred at 78 OC for 30 min, then warm up to room temperature. The mixture was then heated at reflux for 1.5 hour, and stirred at room temperature overnight. The reaction was quenched with saturated aqueous ammonium chloride, and diluted with ethyl acetate. The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 50% ethyl acetate/hexanes to provide the title compound, as a light yellow oil (23.4 mg, 37%). 1 H NMR (CDCI 3 ): 8 7.44 (d, 1H), 5.92 (ddd, 1H), 4.30-4.00 (m, 2H), 3.55 (m, 1H), 2.69 (m, 1H), 2.45 (m, 1H), 2.00 (s, 3H), 1.47 (s, 9H), 1.43 (s, 9H), 1.45 1.00 (m, 4H), 0.91 (m, 6H). MS: (M+H) = 461 -189- WO 99/54299 PCT/US99/07945 F F AcHN. OH 'N HH " O TFA 37B. (±)-(2R,3S,5R,1'S)-2-(1-acetamido-3-methyl)butyl-3-(2,2-difluoro-vin-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. The title compound was prepared according to the method described in Example 33B, substituting (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-(2,2-difluoro-vin-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 (cis-2-chloro-vin-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. 'H NMR (DMSO-d 6 ): 5 8.04 (d, J=7.5Hz, 1H), 4.59 (ddd, 1H), 4.23 (m, 1H), 4.14 (m, 1H), 3.48 (m, 1H), 3.39 (m, 1H), 2.91 (m, 1H), 2.43 (m, 1H), 1.85 (s, 3H), 1.58 (m, 1H), 1.40 (m, 1H), 1.31 (m, 1H), 1.22 (m, 1H), 0.89 (d, J=7.5Hz, 3H), 0.83 (d, J=7.5Hz, 3H). MS: (M+H) = 305 Example 38 (±)-(2R,3R, 5R,1'S)-2-(1 -acetamido-3-methyl)butyl-3-(pyrazol-3-yl)-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt -190- WO 99/54299 PCT/US99/07945 OH AcHN. OtBu N H Boc O 38A. (±)-(2R,3R,5R, I1'S,1 "RS)-1l-t-Butoxycarbonyl-2-(1-acetamido-3 methyl)butyl-3-(1-hydroxy-2-propyn-1 -yl)-pyrrolidine-5-carboxvlic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 4A substituting 2-propynyl magnesium bromide in place of ethyl magnesium bromide and substituting (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-(1 acetamido-3-methyl)butyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester (250 mg, 0.587 mmol) in place of (±)-(2R,3R,5R,1'S)-l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester the crude product was used directly in the next reaction. MS: (M+H)+=453 0O Ac HN. OtBu N H Boc6g 38B. (±)-(2R.3R.5R, 1'S)-l-t-Butoxycarbonyvl-2-(1-acetamido-3-methyl)butyl-3-(1 oxo-2-propyn- 1 -yl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'S)-1-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-( 1 hydroxy-2-propyn-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester was reacted with Jones reagent (3.0 M in acetone, 0.33 mL) in acetone (90 mL) at 0 oC to room temperature for 1 hour. The reaction was diluted with ethyl acetate. The organic -191- WO 99/54299 PCT/US99/07945 layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 50% ethyl acetate/hexanes to provide the title compound, as a white solid (yield: 143 mg, 54%). MS: (M+H) = 451 H NN3 AcHN. OtBu H Boc O 38C. (±)-(2R,3R,5R,1'S)-1l-t-Butoxvcarbonyl-2-(1-acetamido-3-methyl)butyl-3 (pyrazol-3-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-(1 oxo-1-ethynyl)methyl-pyrrolidine-5-carboxylic acid t-butyl ester (140 mg, 0.311 mmol) was reacted with hydrazine monohydrate (0.24 mL, 4.944 mmol) in ethanol (12 mL) at room temperature for 4 hours. The reaction was concentrated in vacuo. The residue was purified by chromatography on silica gel using ethyl acetate to provide the title compound, as a white solid (yield: 131 mg, 91%). MS: (M+H) = 465 -192- WO 99/54299 PCT/US99/07945 HN AcHN. OH 'N K HH 2TFA 38D. (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-methyl)butyl-3-(pyrazol-3-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. The title compound was prepared according to the method described in Example 33B, substituting (±)-(2R,3R,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-(pyrazol-3-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1 'S)-1l-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-(cis-2 chloro-vinyl)-pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (DMSO-d 6 ): 68.13 (d, J=7.5Hz, 1H), 7.65 (d, J=2.2Hz, 1H), 6.20 (d, J=2.2Hz, 1H), 4.39 (m, 1H), 4.25 (m, 1H), 3.94 (m, 1H), 3.56 (q, J=7.5Hz, 1H), 2.62(m, 1H), 2.17 (m, 1H), 1.87 (s, 3H), 1.42 (m, 1H), 1.21 (m, 1H), 1.11 (m, 1H), 0.80 (d, J=6.6Hz, 3H), 0.71 (d, J=6.6Hz, 3H). MS: (M+H)+=309 -193- WO 99/54299 PCT/US99/07945 Example 39 (±)-(2R,3R,5R, 1 'S)-2-(1-acetamido-3-methyl)butyl-3-(isoxazol-3-yl)-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt and (±)-(2R,3R,5R, 1'S)-2-(1-acetamido 3-methyl)butvl-3-(isoxazol-5-yl)-pyrrolidine-5-carboxvlic Acid Trifluoroacetic Acid Salt. AcHN. OtBu AcHN. *N OtBu H Boc OHBoc O 39A. (±)-(2R,3R,5R,1'S)-l-t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 (isoxazol-3-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester and (±)-(2R,3R,5R. 1'S) 1-t-Butoxycarbonyl-2-(1-acetamido-3-methl)butyvl-3-(isoxazol-5-yl)-pyrrolidine-5 carboxylic Acid t-Butyl Ester (±)-(2R,3R,5R, 1'S)-1 -t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-(1 oxo-1-ethynyl)methyl-pyrrolidine-5-carboxylic acid t-butyl ester (31 mg, 0.07 mmol) was reacted with hydroxyamine hydrochloride (4.9 mg, 0.07 mmol) and sodium carbonate (3.7 mg, 0.035 mmol) in ethanol (3 mL) at reflux for 30 hours. The reaction was concentrated in vacuo. The residue was purified by chromatography on silica gel using 3% methanol/dichloromethane to provide the title compound, as an oil (yield: 11.5 mg, 36%). MS: (M+H)*= 466 -194- WO 99/54299 PCT/US99/07945 0)0 AcHN. OH AcHN. OH • O O 2TFA 2TFA 39B. (±)-(2R.3R,5R,1'S)-2-(1-acetamido-3-methyl)butyl-3-(isoxazol-3-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt and (±)-(2R,3R,5R, 1'S)-2 (1-acetamido-3-methyl)buyl-3-(isoxazol-5-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. The title compound was prepared according to the method described in Example 33B, substituting (±)-(2R,3R,5R, 1 'S)-l-t-butoxycarbonyl-2-(1-acetamido 3-methyl)butyl-3-(isoxazol-3-yl)-pyrrolidine-5-carboxylic acid t-butyl ester and (±) (2R,3R,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-(isoxazol-5 yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1-t butoxycarbonyl-2-(1 -acetamido-3-methyl)butyl-3-(cis-2-ch loro-vinyl)-pyrro lidine-5 carboxylic acid t-butyl ester. 1 H NMR (DMSO-d 6 ): 8 8.91, 8.54 (d, 1H), 8.12, 8.05 (d, J=7.5Hz, 1H), 6.64, 6.43 (d, 1H), 4.48, 4.51 (m, 1H), 4.28 (m, 1H), 3.97, 3.89 (m, 1H), 3.70, 3.81 (m, 1H), 2.72 (m, 1H), 2.20, 2.25 (m, 1H), 1.83, 1.80 (s, 3H), 1.48 (m, 1H), 1.34 1.10 (m, 2H), 0.83, 0.84 (d, J=6Hz, 3H), 0.77, 0.78 (d, J=6Hz, 3H). MS: (M+H) =310 Example 40 -195- WO 99/54299 PCT/US99/07945 (±)-(2R,3S. 5R, 1'R)-2-(1 -Acetamido-2-hydroxy)ethyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt AcO-, Z OtBu Ph 40A. (±)-(2R,3R,5R)-1l-Benzyl-2-vinyl-3-(acetoxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R)-1 -Benzyl-2-vinyl-3-(hydroxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (54.2 g, 0.17 mol) and 4-(dimethylamino)pyridine (0.5 g, 4.1 mmol), in anhydrous pyridine (400 mL) was reacted with acetic anhydride (30 mL, 0.32 mol) at 00C for 1 hour then allowed to warm to room temperature. The reaction was stirred an additional 16 hours. The pyridine was removed in vacuo at 300C. The residue was partitioned between ethyl acetate (100 mL) and of water (400 mL). The aqueous layer was extracted with ethyl acetate (3 x 100 mL) and the combined ethyl acetate layers were washed with brine, dried with MgSO 4 , filtered, and concentrated. The crude product was purified by chromatography on silica gel using 10% ethyl acetate/hexanes to provide the title compound as a colorless oil (yield: 49.6 g, 81%). 1 H NMR (CDC1 3 ) 8 7.28 (m, 4H), 7.21 (m,1H), 5.68 (m,1H), 5.21 (m, 2H), 4.16 (dd, J=6.3, 10.7 Hz, 1H), 4.10 (dd, J=7.3, 10.7 Hz, 1H), 3.92 (d, J=13.7 Hz, 1H), 3.64 (d, J=13.7 Hz, 1H), 3.52 (m, 1H), 3.50 (m, 1H), 2.33 (m, 1H), 2.26 (m, 1H), 2.02 (s, 3H), 1.62 (m, 1H), 1.45 (s, 9H). MS (M+H) = 360. -196- WO 99/54299 PCT/US99/07945 AcO HO N OIBu
N
o HO O Ph 40B. (±)-(2R,3R,5R,1'R)-and (±)-(2R,3R,5R,1'S)-I-Benzyl-2-(1,2 dihydroxy)ethyl-3-(acetoxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R)-l1-benzyl-2-vinyl-3-(acetoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester.(52.5 g, 0.15 mol) and 4-methylmorpholine N-oxide (54.7 g, 0.47 mol) in acetone (540 mL) and water (60 mL) was reacted with osmium tetroxide (200 mg, 0.8 mmol). After 24 hours, the reaction was quenched with 10% sodium thiosulfate (250 mL) concentrated in vacuo. The aqueous layer was extracted with ethyl acetate (3 x 300 mL) and the combined organic layers were washed with brine, dried over MgSO 4 , filtered and concentrated. The residue was purified by chromatography on silica gel using a gradient elution of ethyl acetate and dichloromethane to provide the title compound as a viscous oil (yield: 41.2 g, 72%). 1 H NMR (DMSO) 8 7.32 (m, 3H), 7.30 (m, 1H), 7.22 (m, 1H), 4.48 (t, J=5.4 Hz, 1H), 4.42 (d, J=5.4 Hz, 1H), 4.04 (m, 1H), 4.01 (m, 1H), 3.97 (m, 1H), 3.80 (d, J=13.2 Hz, 1H), 3.78 (m, 1H), 3.43 (m, 1H), 3.39 (m, 1H), 3.32 (m, 1H), 3.07 (t, J=4.9 Hz,IH), 2.48 (m,1H), 2.19 (m, 1H), 1.99 (s, 3H), 1.57 (dt, J=13.7, 2.0 Hz, 1H), 1.38 (s, 9H). MS (M+H) = 394. -197- WO 99/54299 PCT/US99/07945 AcO, HO OtBu HO OH 0 40C. (±)-(2R.3R,5R,1'R) and (±)-(2R,3R,5R,1'S)-2-(1,2-Dihydroxy)ethyl-3 (acetoxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'R) and (±)-(2R,3R,5R,1'S)-l-Benzyl-2-(1,2 dihydroxy)ethyl-3-(acetoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (24 g, 61 mmol) in ethanol (300 mL) was reacted with ammonium formate (38.5 g, 0.61 mol) and 10% Pd/C (2g) for 2 hours at reflux. The reaction was cooled and the catalyst removed by filtration through Celite. The filtrate was concentrated in vacuo to provide the title compound (yield: 16.7 g, 90%). 1 H NMR (DMSO) 8 4.56 (m, 1H), 4.30 (m, 1H), 4.06 (dd, J=5.8, 10.9 Hz, 2H), 3.79 (dd, J=8.8, 10.5 Hz, 2H), 3.49 (m, 4H), 3.00 (m, 1H), 2.35 (m, 1H), 2.16 (dt, J=12.6, 8.5 Hz, 1H), 2.01 (s, 3H), 1.52 (m, 1H), 1.40 (s, 9H). MS (M+H) = 304. AcO-, HO OtBu H Bocu HO 40D. (±)-(2R,3R,5R,1'R) and (±)-(2R,3R,5R,1 'S)-l-t-Butoxycarbonvl 2-(1,2 dihydroxy)ethyl-3-(acetoxvmethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R, 1 'R) and (±)-(2R,3R,5R, 1 'S)-2-(1,2-Dihydroxy)ethyl-3 (acetoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (33.4 g, 0.11 mol) in methanol (250 mL) and water (50 mL) was reacted with di-t-butyl dicarbonate (33.6 g, 0.15 mol) for 48 hours at room temperature. The methanol was removed in vacuo and the residue diluted with water ( 500 mL), and extracted with ethyl acetate (3 x 200 mL). The combined ethyl acetate layers were washed with brine, -198- WO 99/54299 PCT/US99/07945 dried with MgSO 4 , filtered and concentrated. The residue was chromatographed on silica gel using methanol/dichloromethane to provide the title compound as a white solid (yield: 32.8 g, 78%) 1 H NMR (DMSO) 6 4.80 (m, 1H), 4.45 (m, 1H), 4.08 (m, 1H), 3.91 (m, 2H), 3.82 (m, 1H), 3.71 (m, 1H), 3.28 (m, 2H), 2.48 (m, 1H), 2.07 (m, 2H), 2.01 (m, 3H), 1.39 (m, 18H). MS (M+H) = 404. AcO HO OtBu H Boc u TIPSO O 40E (±)-(2R,3R,5R,1'R) and (±)-(2R,3R,5R.,1'S)-l-t-Butoxycarbonyl 2-(1 hydroxy-2-triisopropylsilyloxy)ethyl-3-(acetoxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'R) and (±)-(2R,3R,5R,1'S)-l-t-Butoxycarbonyl 2-(1,2 dihydroxy)ethyl-3-(acetoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester.(26.5 g, 66 mmol) in anhydrous dimethylformamide (200 mL ) was reacted with imidazole (8.9 g, 0.13 mol) and triisopropylsilyl chloride (19.0 g, 99 mmol) for4 hours at room temperature. The solvent was removed under vacuum and the residue partitioned between 300 mL of water and 150 mL of ethyl acetate. The aqueous layer was extracted with ethyl acetate (2 x 100 mL), and the combined ethyl acetate layers extracted with brine, dried with MgSO 4 , filtered and concentrated. The residue was purified by chromatography on silica gel using 10% ethyl acetate/hexanes to provide the title compound as a colorless oil (yield: 28.9 g, 79%). -199- WO 99/54299 PCT/US99/07945 1 H NMR (CDCI 3 ) 8 4.22 (m, 1H), 4.04 (m, 3H), 3.87 (t, J=2.0 Hz, 1H), 3.74 (dd, J=4.9, 9.8 Hz, 1H), 3.58 (dd, J=7.8, 10.2 Hz, 1H), 3.39 (bs, 1H), 2.61 (m, 2H). 2.03 (s, 3H), 1.75 (m, 1H), 1.46 (m, 18H), 1.07 (m ,18H). MS (M+H) + = 560. AcO O n OtBu H o TIPSO 40F (±)-(2R,3R,5R)-1l-t-Butoxycarbonyl-2-(1-oxo-2-triisopropylsilyloxy)ethyl-3 (acetoxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. Dimethylsulfoxide (6 mL, 85 mmol) was added slowly to a solution of oxalyl chloride (2 M) (19.3 mL, 38.6 mmol) in dry dichloromethane (70 mL) at -78 0 C. After 10 minutes, a solution of (±)-(2R,3R,5R,1'R) and (±)-(2R,3R,5R,1'S)-1-t butoxycarbonyl 2-(1-hydroxy-2-triisopropylsilyloxy)ethyl-3-(acetoxymethyl) pyrrolidine-5-carboxylic acid t-butyl ester (14.4 g, 26 mmol) in dry dichloromethane (75 mL) was added at a rate such that the temperature did not exceed -70 0 C. After 1.5 hours, triethylamine (18 mL, 0.13 mol) was added and the temperature allowed to rise to 0 0 C. The reaction was quenched with a solution of ammonium chloride, diluted with water, and extracted with dichloromethane (3 x 100 mL). The combined dichloromethane layers were extracted with brine, dried with MgSO 4 , filtered and concentrated. The residue was purified by chromatography on silica gel using 10% ethyl acetate/hexanes to provide the title compound as a colorless oil: (yield: 11 g, 77%). 1 H NMR (CDCI 3 ) 8 4.32 (m, 6H), 2.43 (m, 2H), 2.04 (s. 3H), 1.78 (m, 1H), 1.48 (s, 9H), 1.41 (s, 9H), 1.1 (m, 21H). MS (M+H) = 558. -200- WO 99/54299 PCT/US99/07945 AcO-,
H
2 N OtBu B O TIPSO 40G (±)-(2R,3R,5R, 1'R) and (±)-(2R,3R,5R, 1l'S)-1-t-Butoxycarbonyl 2-(1-amino 2-triisopropylsilyloxy)ethyl-3-(acetoxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R)-1l-t-Butoxycarbonyl 2-(1-oxo-2-triisopropylsilyloxy)ethyl-3 (acetoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester.(22 g, 39 mmol) in methanol (1 L) was reacted with ammonium acetate (77 g, 1.0 mol) and sodium cyanoborohydride (24.8 g, 0.39 mol) at reflux for 2 hours. The solvent was removed under in vacuo, and the residue was partitioned between water (300 mL) and dichloromethane (300 mL). The aqueous layer was extracted with dichloromethane (2 x 100 mL) and the combined organic layers were washed with brine, dried with MgSO 4 , filtered and concentrated to provide the title compound (crude yield: 22.0g, 100%). AcO-, AcO-, AcHN, OtBu AcHN OtBu TIPSO O TIPSO O 40H (±)-(2R,3R.5R,1'R) and (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-(1 acetamido-2-triisopropylsilylox(y)ethyl-3-(acetoLxymethyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R, 1 'R) and (±)-(2R,3R,5R, 1'S)-1 -t-Butoxycarbonyl-2-(1-amino 2-triisopropylsilyloxy)ethyl-3-(acetoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (approx 39 mmol) in dichloromethane (500 mL) was reacted with acetic anhydride (18 mL, 0.19 mol), triethylamine (27.5 mL, 0.20 mol) and -201- WO 99/54299 PCT/US99/07945 dimethylaminopyridine (50 mg, 0.39 mmol) for 18 hours at room temperature. The reaction was quenched with a solution of ammonium chloride. The aqueous layer was extracted with dichloromethane (3 x 100 mL) and the combined organic layers extracted with brine, dried with MgSO 4 , filtered, and concentrated. The residue was chromatographed on silica gel using ethyl acetate/hexanes to provide the title compound (±)-(2R,3R,5R, 1'R)-1l-t-butoxycarbonyl 2-(1 acetamido-2-triisopro pylsilyloxy)ethyl-3-(acetoxymethyl)-pyrrolidine-5-carboxyl ic acid t-butyl ester (9.14 g, 39%) and (±)-(2R,3R,5R,1'S)-1-t-butoxycarbonyl 2-(1 acetamido-2-triisopropylsilyloxy)ethyl-3-(acetoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester (9.75 g, 41% ) as white solids. (±)-(2R,3R,5R,1'R) 1 H NMR (CDCl 3 ) 8 7.38 (d, J=8.3 Hz, 1H), 4.34 (m, 1H), 4.20 (dd, J=2.4, 10.3 Hz, 1H), 4.09 (dd, J=8.8, 10.2 Hz. 1H), 4.02 (dd, J=7.3, 10.1 Hz, 1H), 3.88 (m, 1H), 3.71 (dd, J=4.4, 10.3 Hz, 1H), 3.65 (dd, J=7.9, 10,3 Hz, 1H), 2.74 (m, 1H), 2.60 (m, 1H), 2.04 (s, 3H), 1.98 (s, 3H), 1.69 (dt, J=14.1, 2.5 Hz, 1H), 1.46 (s, 9H), 1.42 (s, 9H), 1.07 (m, 21H). MS (M+H) = 601 (±)-(2R,3R,5R,1'S) 1 H NMR (CDCI 3 ) 6 6.82 (d, 1H), 4.10 (m, 4H), 3.81 (m, 3H), 2.55 (m, 2H), 1.98 (m, 7H), 1.46 (s, 9H), 1.42 (s, 9H), 1.07 (m, 21H). MS (M+H)* = 601. -202- WO 99/54299 PCT/US99/07945 HO-, AcHN, OtBu H Boc TIPSO O 401 (±)-(2R.3 R,5R, 1'R)-l-t-Butoxycarbonyl 2-(1-acetamido-2 triisopropylsilyloxy)ethyl-3-hydroxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R, 1 'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-2 triisopropylsilyloxy)ethyl-3-(acetoxymethyl)-pyrrolidine-5-carboxylic acid t-butyl ester.(8.2 g, 13.66 mmol) in methanol (200 mL) and water (50 mL) was reacted with potassium carbonate (19 g, 136 mmol) at room temperature for 2 hr. The solvent was then removed in vacuo and the residue was partitioned between water (100 mL) and dichloromethane (3 x 100 mL). The organic extracts were dried over magnesium sulfate, filtered and concentrated in vacuo to provide the title compound as a colorless oil. H 0=K AcHN OtBu N H oc TIPSO O 40J (±)-(2R.,3R,5R, 1'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-2 triisopropylsilyloxy)ethyl-3-formyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 2A substituting (±)-(2R,3R,5R,1'R)-1l-t-butoxycarbonyl 2-(1-acetamido (2-triisopropylsilyloxy)ethyl-3-hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3 hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 5.9 g, 78%). -203- WO 99/54299 PCT/US99/07945 1 H NMR (CDCI 3 ) 8 1.04 -1.07 (m, 21H), 1.42 (s, 9H), 1.43 (s, 9H), 1.99 (s, 3H), 2.42 (m, 1H), 2.62 (m, 1H), 3.04 (m, 1H), 3.69 (m, 1H), 3.82 (m, 1H), 4.08 (m, 1H), 4.38 (m, 1H), 4.57 (t, 1H), 7.33 (br d, 1H), 9.65 (s, 1H). MS: (M+H) = 557. CH AcHI OtBu N TIPSO O 40K (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-2 triisopropylsilyloxy)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 35A substituting (±)-(2R,3R,5R,1'R)-1l-t-butoxycarbonyl 2-(1-acetamido 2-triisopropylsilyloxy-)ethyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-l1-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 formyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 5.9 g, 78%). 1 H NMR (CDCl 3 ) 8 1.03 -1.10 (m, 21H), 1.44 (s, 9H), 1.47 (s, 9H), 1.55 (m,1H), 1.64 (dd, 3H), 1.96 (s, 3H), 2.55 (m, 1H), 3.42 (m, 1H), 3.62 - 3.71 (m, 3H), 4.20 (dd, 1H), 4.30 (m, 1H), 5.39 (m, 1H), 5.48 (m, 1H), 7.73 (br d, 1H). MS: (M+H) = 569 -204- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OtBu H Boc"O HOO 40L (±)-(2R.,3S,5R,1'R)-1-t-Butoxycarbonyl 2-(1-acetamido-2-hydroxy-)ethyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, I 'R)-1-t-Butoxycarbonyl 2-(1-acetamido-2 triisopropylsilyloxy)ethyl-3-(cis-propen- I -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (4.85 g, 8.54 mmol) in THF (100 mL) was reacted with tetrabutyl ammonium fluoride (1IM in THF) (12.8 mL, 12.8 mmol) for 30 minutes at room temperature. Water (100 mL) was added followed by extraction using dichloromethane (2 x 100 mL). This organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 2/1: ethyl acetate/hexane to provide the title compound as a colorless solid (yield: 3.1g, 89%). 'H NMR (CDCI 3 ): d 1.44 (s, 9H), 1.47 (s, 9H), 1.56 (dd, 3H), 1.80 (m, IH), 2.02 (s, 3H), 2.67 (m, 1H), 3.11 (t, 3H), 3.44 (dd, 1H), 3.59 (dd, 1H), 3.74-3.84 (m, 2H), 4.15 (dd, 1H) 5.39 (m, 1H), 5.58 (m, 1H), 6.42 (brd, 1H). MS: (M+H) = 413.
CH
3 AcHN, OH HO 0 TFA 40M (±)-(2R.3S,5R, 1'R)-2-(1-Acetamnido-2-hydroxy)ethyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 33B, substituting (±)-(2R,3S,5R,1'R)-1-t-butoxycarbonyl-2-(1-acetamido -205- WO 99/54299 PCT/US99/07945 2-hydroxy)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 (trans-2-chloro-vinyl)-pyrrolidine-5-carboxyic acid t-butyl ester (yield: 18.0 mg, 100%). 1 H NMR (DMSO-d 6 ): d 1.66 (dd, 3H), 1.71 (dt, 1H), 1.87 (s, 3H), 2.41 (dt, 1H), 3.18 (m, 1H), 3.43 (dd, 1H), 3.61 (m, 1H), 4.13 (m, 1H), 4.35 (m, 1H), 5.25 (m, 1H), 5.51 (m, 1H), 8.05 (d, 1H), 9.16 (brs, 2H). MS: (M+H) = 257. Example 41 (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt CH AcH OtBu HBoc O 0OH 41A (±)-(2R,3S.5R,1'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-1l-formyl)methyl-3 (cis-propen-1 -yli)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1 'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-2-hydroxy)ethyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester.(600 mg, 1.46 mmol) in dichloromethane (50 mL) was reacted with Dess-Martin Periodinane (928 mg, 2.18 mmol) for 1 hour at room temperature. The reaction was quenched with 1M aqueous sodium thiosulfate (50 mL), stirred for 20 minutes then extracted with dichloromethane (3 x 100 mL). The organic layer was dried over magnesium sulfate, concentrated in vacuo. The residue was purified by column -206- WO 99/54299 PCT/US99/07945 chromatography on silica gel using 2/1: ethyl acetate/hexane to provide the title compound (yield: 547 mg, 92%). 1H NMR (CDCI 3 ) d 9.40 (d, J= 1 Hz, 1H), 7.88 (bd), 5.69 (m, 1H), 5.27 (m, 1H), 4.78 (dd, J= 9.5, 1. Hz, 1H), 4.21 (t, J= 8. Hz, 1H), 3.45 (m, 2H), 2.41 (m, 1H), 2.09 (s, 3H), 1.69 (dd, J= 7.0, 1. Hz, 3H), 1.55 (m, 1H), 1.46 (s, 9H), 1.40 (s, 9H). MS: (M+H) = 411, (M-H)- = 409.
CH
3
CH
3 AcHN. OtBu AcHN OBu
H
3 C OH BOC H 3 C H BOC OH H 41B (±)-(2R,3S,5R,1'R.2'R) and (±)-(2R,3S,5R, 1'R,2'S)-1-t-Butoxycarbonyl-2 (1-acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t Butyl Ester. (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-1l-formyl)methyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (780 mg, 1.90 mmol) in THF (20 mL) was added dropwise to a solution of ethylmagnesium bromide (3M in ether) (3.17 mL, 9.51 mmol) in THF (15 mL) at room temperature and reacted for 40 minutes. The reaction was quenched with water (20 mL) and saturated aqueous ammonium chloride (20 mL) followed by extraction using dichloromethane (3 x 50 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 2/1: ethyl acetate/hexane to provide the title compounds (±)-(2R,3S,5R, 1 'R,2'R)-1l-t-butoxycarbonyl 2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 472 mg, 56%) and (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl 2-(1-acetamido-2 -207- WO 99/54299 PCT/US99/07945 hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester(yield: 82 mg, 10%) as a colorless oils. (±)-(2R,3S,5R,1'R,2'R) = MS: (M+H) = 441, (M+Na)+= 463, (M-H)- = 439. (±)-(2R,3S,5R,1'R,2'S) = MS: (M+H) = 441, (M+Na)+= 463, (M-H)- = 439.
CH
3 AcHN. OH 'N fi H 0 OH TFA 41C (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)butyl-3-(cis-propen-1 vl)-pyDvrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt (±)-(2R,3S,5R, 1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (300 mg, 0.68 mmol) was reacted with trifluoroacetic acid (8 mL) in dichloromethane (2 mL) at room temperature for 6 hrs. The reaction was concentrated in vacuo overnight to provide the title compound (yield: 311 mg) as a colorless solid. 1 H NMR (500 MHz, DMSO-d 6 ) 8:7.89 (d, J= 8.7 Hz, 1H), 5.48 (m, 1H), 5.29 (m, 1H), 4.30 (m, 1H), 4.02 (m, 1H), 3.73 (m, 1H), 3.43 (m, 1H), 3.15 (m, 1H), 2.41 (m, 1H), 1.82 (s, 3H), 1.63 (m, 1H), 1.59 (dd, J = 6.8, 1.9 Hz, 3H), 1.55 (m, 1H), 1.27 (m, 1H), 0.85 (t, J = 7.3 Hz, 3H). MS: (M+H) = 285, (M+Na)*= 307, (M-H)- = 283. -208- WO 99/54299 PCT/US99/07945 Example 42 (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN OtBu N H Bo O 0O 42A (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-oxo)butvl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester.(460 mg, 1.05 mmol) was reacted with Dess-Martin Periodinane (666 mg, 1.57 mmol) in dichloromethane (30 mL) at room temperature for 17 hours. The reaction was quenched with 1M aqueous sodium thiosulfate (50 mL and stirred for 20 minutes. The reaction was was extracted with dichloromethane (3 x 100 mL). The organic layer was dried over magnesium sulfate,filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 2:1: ethyl acetate/hexane to provide the title compound as a colorless semi-solid (yield: 440 mg, 96%). MS: (M+H) = 439, (M+Na)+= 461, (M-H)- = 437. -209- WO 99/54299 PCT/US99/07945
CH
3
CH
3 AcHN. OtBu AcHN OtBu HoOCo HNO o OH OH 42B (±)-(2R.3S,5R,1'R,2'R) and (±)-(2R,3S,5R,1'R,2'S)-1-t-Butoxycarbonyl-2 (1-acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t Butyl Ester. (±)-(2R,3S,5R, 1 'R)-1l-t-Butoxycarbonyl-2-(1 -acetamido-2-oxo)butyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (435 mg, 0.99 mmol) in methanol (30 mL) was reacted with sodium borohydride (188 mg, 4.97 mmol) at room temperature for 0.5 hours. The solvent was removed in vacuo and water (30 mL) was added. The aqueous layer was extracted with dichloromethane (3 x 50 mL). This organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 2:1 ethyl acetate/hexane to provide the title compounds (±) (2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester.(yield: 305 mg, 70%) and compounds (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 17 mg, 4%). -210- WO 99/54299 PCT/US99/07945
CH
3 AcHN- OH HH OH TFA 42C (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (300 mg, 0.68 mmol) was reacted with trifluoroacetic acid (8 mL) in dichloromethane (2 mL) at room temperature for 6 hrs. The reaction was concentrated in vacuo overnight and triturated with acetonitrile (2 x 5 mL) to provide the title compound (yield: 311 mg) as a colorless solid. 1 H NMR (500 MHz, DMSO-d 6 ) 5: 7.89 (d, J = 8.7 Hz, 1H), 5.48 (m, 1H), 5.29 (m, 1H), 4.30 (m, 1H), 4.02 (m, 1H), 3.73 (m, 1H), 3.43 (m, 1H), 3.15 (m, 1H), 2.41 (m, 1H), 1.82 (s, 3H), 1.63 (m, 1H), 1.59 (dd, J = 6.8, 1.9 Hz, 3H), 1.55 (m, 1H), 1.27 (m, 1H), 0.85 (t, J = 7.3 Hz, 3H). MS: (M+H)*= 285, (M+Na)+= 307, (M-H)- = 283. -211- WO 99/54299 PCT/US99/07945 Example 43 (±)-(2R, 3S, 5R R,R,2'R)-2-(1 -Acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt TI-'
CH
3 AcHN. OH N H 0 OH TFA The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1 -t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: .0065 g, 100%). 1 H NMR (DMSO-d 6 ) 8 7.90 (d, J=8.8Hz, 1H), 5.47 (m, 1H), 5.29 (t, J=9.8 Hz, 1H), 4.29 (t, J=8.8Hz, 1H), 4.02 (q, J=6.8Hz, 1H), 3.71 (bt, J=8Hz, 1H), 3.43 (m, 1H), 3.15 (quint., J=8.8Hz, 1H), 2.41 (dt, J=12.7,7.8Hz, 1H), 1.82 (s, 3H), 1.64 (m, 1H), 1.58 (dd, J=6.8,1.5Hz, 3H), 1.53 (m, 1H), 0.85 (t, J=7.3Hz, 3H). MS: (M+H) = 285, (M+Na)+ = 307, (M-H)- = 283, (M+CF 3 COOH)- = 397, (2M-1) = 563 Example 44 -212- WO 99/54299 PCTIUS99/07945 (±)-(2R, 3S,5R. 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-cyano)propyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt CH3 CH 3 AcHN. OtBu AcHN. OtBu N I N N-C H Boc O N=C H Boc OH OH 44A (±)-(2R.3S,5R,1'R,2'R) and (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2 (1-acetamido-2-hydroxy-3-cyano)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl 2-(1 -acetamido-1 -formyl)methyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (150 mg, 0.37 mmol) in THF (10 mL) was added dropwise to a solution of the lithium enolate of acetonitrile (1.83 mmol, 5 equivalents) in THF (15 mL) at -78 oC and reacted for 15 minutes. The reaction was quenched with saturated aqueous ammonium chloride (10 mL) and water (1OmL) followed by extraction using dichloromethane (2 X 50 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 2/1: ethyl acetate/hexane to provide the title compounds (±) (2R,3S,5R, 1 'R,2'R)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy-3-cyano)propyl 3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 95mg, 58%) and (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy-3 cyano)propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester(yield: 30 mg, 18%) as a colorless oils. (±)-(2R,3S,5R,1'R,2'R) = MS: (M+H)*=452, (M-H) =450 (±)-(2R,3S,5R,1'R,2'S) ='H NMR (CDCl 3 ) 8 8.14 (d, J=8.9Hz, 1H), 5.51 (m, 1H), 5.38 (m, 1H), 4.25 (m, 1H), 4.19 (m, 1H), 3.94 (m, 1H), 3.74 (m, 1H), 3.22 -213- WO 99/54299 PCT/US99/07945 (m, 1H), 2.54 (m, 1H), 2.47 (m, 2H), 2.04 (s, 3H), 1.69 (m, 1H),1.65 (dd, J=6.5, 1.8Hz, 3H), 1.47 (s, 9H), 1.45 (s, 9H) MS: (M+H) =452, (M-H)- =450
CH
3 AcHN. "OH NEC HH O OH TFA 44B (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy-3-cvano)propyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-cyano)propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 4.5 mg, 95%%). 1 H NMR (DMSO-d 6 ) 8 7.98 (d, J=10.0 Hz, 1H), 5.49 (m, 1H), 5.27 ( m, 1H), 4.30 (m, 1H), 4.15 (m, 1H), 3.75(m, 1H), 3.18 (m, 1H), 2.72-2.58 (m, 2H), 2.41 (m, 1H), 1.85 (s, 3H), 1.65 (m, 1H), 1.61 (dd, J=6.70, 1.80 Hz, 3H) MS: (M+H) =296, (M-H)-=294 Example 45 -214- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R, 1'R,2'R)-2-(1-Acetamido-2-hydroxy-3-cvano)propyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. , N H N=-C H0 N-EAC OH OH TFA The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-cyano)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 8 mg, 95%). H NMR (DMSO-d6) 8 7.75(d, J=9.0 Hz, 1H), 5.47(m, 1H), 5.25(m, 1H), 4.46(m, 1H), 4.20(m, 1H), 4.13(m, 1H), 3.56(m, 1H), 3.15(m, 1H), 2.55(m, 2H), 2.42(m, 1H), 1.82(s, 3H), 1.72(m, 1H), 1.55(dd, J=6.71, 1.83, 3H) MS: (M+H) =296, (M+23)+ =318, (M-H) =294 Example 46 -215- WO 99/54299 PCT/US99/07945 (±)-(2R,3S, 5R. 1'R,2'R)-2-(1 -Acetamido-2-hydroxy-3-ethoxycarbonyl)propyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 CH 3 AcHN. .OtBu AcHN. OtBu Soc o Boc OH OH EtO 0O EtO 0 46A (±)-(2R,3S,5R,1'R,2'R) and (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxvcarbonyl-2 (1 -acetamido-2-hydroxy-3-ethoxycarbonyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine 5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-1 -formyl)methyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (900 mg, 2.187 mmol) in THF (40 mL) was added dropwise to a solution of the lithium enolate of ethyl acetate (8.75 mmol, 4 equivalents) in THF (40 mL) at -78 OC and reacted for 15 minutes. The reaction was quenched with saturated aqueous ammonium chloride followed by extraction using dichloromethane (3 X). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1:1 ethyl acetate/hexane to provide the title compounds (±)-(2R,3S,5R, 1'R,2'R)-1 -t butoxycarbonyl-2-(1-acetamido-2-hydroxy-3-ethoxycarbonyl)propyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 690 mg, 63%) and (±) (2R,3S,5R,1'R,2'S)-1 -t-butoxycarbonyl-2-(1 -acetamido-2-hydroxy-3 ethoxycarbonyl)propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 246 mg, 22.5%). (±)-(2R,3S,5R,1'R,2'R) 1 H NMR (CDCI 3 ): 6 5.99 (d, 1H), 5.60 (m, 1H), 5.36 (m, 1H), 4.81 (m, 1H), 4.15 (m, 4H), 3.74 (m, 1H), 3.07 (m, 1H), 2.68 (m, -216- WO 99/54299 PCT/US99/07945 1H), 2.48 (m 1H), 2.33 (m, 1H), 2.03 (s, 3H), 1.54 (dd, 3H), 1.47 (s, 9H), 1.46 (s, 9H), 1.24 (t, J=7.5Hz, 3H). MS: (M+H)+=499 (±)-(2R,3S,5R,1'R,2'S) 1 H NMR (CDCI 3 ): 8 7.93 (d, 1H), 5.44 (m, 2H), 4.19 (m, 4H), 4.03 (m, 1H), 3.72 (m, 1H), 3.37 (m, 1H), 2.63 (m. 1H), 2.48 (m, 2H), 2.01 (s, 3H), 1.65 (dd, 3H), 1.48 (s, 9H), 1.46 (s, 9H), 1.26 (t, J=7.5Hz, 3H). MS: (M+H)+=499
CH
3 AcHN. OH ' N OH 0 EtO O TFA 46B (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-3 ethoxycarbonyl)propyl-3-(cis-propen- 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-ethoxycarbonyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester. 1 H NMR (DMSO-d 6 ): 8 7.74 (d, J=9Hz, 1H), 5.48 (m, 1H), 5.25 (m, 1H), 4.43 (m, 1H), 4.24 (m, 1H), 4.14 (m, 1H), 4.06 (q, J=7.5Hz, 2H), 3.54 (m, 1H), 3.16 (m, 1H), 2.41 (m, 1H), 2.36 (m, 2H), 1.82 (s, 3H), 1.77 (m, 1H), 1.56 (dd, 3H), 1.18 (t, J=7.5Hz, 3H). -217- WO 99/54299 PCT/US99/07945 MS: (M+H) = 343 Example 47
CH
3 AcHN. OH N HHO OH STFA EtO 0 (±)-(2R.3S. 5R,1 'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-ethoxcarbonyl)propyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound is prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-ethoxycarbonyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester. 1 H NMR (DMSO-d 6 ): 6 7.93 (d, J=9Hz, 1H), 5.48 (m, 1H), 5.30 (m, 1H), 4.19 (m, 1H), 4.09 (m, 1H), 4.06 (q, J=7.5Hz, 2H), 3.94 (m, 1H), 3.73 (m, 1H), 3.18 (m, 1H), 2.54 (dd, 1H), 2.40 (m, 1H), 2.27 (m, 1H), 1.82 (s, 3H), 1.65 (m, 1H), 1.60 (dd, 3H), 1.19 (t, J=7.5Hz, 3H). MS: (M+H)*=343 -218- WO 99/54299 PCT/US99/07945 Example 48
CH
3 AcHN. K OH ' N ' HH
CH
3 OH TFA (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)propyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0030 g, 100%). 1 H NMR (DMSO-de) d 8.97 (bs, 1H), 7.88 (d, J=8.5 Hz, 1H), 5.45 (m, 1H), 5.28 (t, J = 9.1Hz, 1H), 4.30 (t, J=8.6Hz, 1H), 3.94 (q, J=7.3Hz, 1H), 3.71 (t, J=8.0Hz, 1H), 3.62 (m, 1H), 3.15 (quint., J=9.0Hz, 1H), 2.40 (dt, J=12.8, 7.6Hz, 1H), 1.83 (s, 3H), 1.65 (m, 1H), 1.59 (dd, J=7.0, 1.5Hz, 3H), 1.08 (d, J=5.5Hz, 3H). MS: (M+H) = 271, (M+Na)+ = 293, (M-H) = 269. -219- WO 99/54299 PCT/US99/07945 Example 49
CH
3 AcHN. OH
CH
3 'OH TFA (±)-(2R, 3S, 5R, 1'R.2'R)-2-(1 -Acetamido-2-hydroxy) propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0143 g, 100%). 1 H NMR (DMSO-d 6 ) 8 7.70 (d, J=9.1 Hz, 1H), 5.49 (m, 1H), 5.25 (t, J = 9.1Hz, 1H), 4.43 (t, J=8.6Hz, 1H), 4.03 (m, 1H), 3.92 (m, 1H), 3.55 (t, J=8.5Hz, 1H), 3.17 (quint., J=8.5Hz, 1H), 2.42 (dt, J=12.8,7.3Hz, 1H), 1.85 (s, 3H), 1.72 (dt, J=12.8,10.0Hz, 1H), 1.57 (dd, J=6.7,1.8Hz, 3H), 1.04 (d, J=6.1Hz, 3H). MS: (M+H)
+
= 271, (M+Na) = 293, (M-H)- = 269 -220- WO 99/54299 PCT/US99/07945 Example 50 (±)-(2R, 3S5 R, 1 'R,2'S)-2-( 1 -Acetamido-2-hydroxy-2-vinyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. CH3 )-mCH3 Bo AcHN O tB u A . .HN. OtBu AcH.N OH OH 50A (±)-(2R.3S,5R,1'R,2'S) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2 (1-acetamido-2-hydroxy-2-vinvl)ethyl-3-(cis-propen-1-yvl)-pyrrolidine-5-carbonxyvlic Acid t-Butyl Ester. The title compounds were prepared according to the method described in Example 41B, substituting vinyl magnesium bromide for ethyl magnesium bromide to provide (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy-2-vinyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 6.5 mg, 18%) and (±)-(2R,3S,5R,1'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 22 mg, 59%). (±)-(2R,3S,5R,1'R,2'S) MS: (M+H) =439, (M-H) = 437 (±)-(2R,3S,5R,1'R,2'R) MS: (M+H) =439, (M-H)- = 437 -221- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH ' N S HH " H TFA 50B (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy-2-vinyl)ethyl-3-(cis propen-1 -ylv)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-vinyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 5 mg, 96%). 1 H NMR (DMSO-d 6 ) 87.85 (d, J=9.1 Hz, 1H), 5.76 (m, 1H), 5.47(m, 1H), 5.25 (m, 2H), 5.14 (m, 1H), 4.29 (m, 1H), 4.05 (m, 1H), 3.96 (m, 1H), 3.71 (m,1H), 3.18 (m, 1H), 2.41 (m, 1H), 1.78 (s, 3H), 1.64 (m, 1H), 1.59 (dd, J=6.71, 1.21Hz, 3H) MS: (M+H) =283, (M+23) =305, (M-H) =281 -222- WO 99/54299 PCT/US99/07945 Example 51
CH
3 AcHN. OH OH TFA (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-2-vinyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-vinyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 6 mg, 95%). 1 H NMR (DMSO-d 6 ) 5 7.84 (d, J=9.7Hz, 1H), 5.78 (m, 1H), 5.48 (m, 1H), 5.23 (m, 34.43 (m, 1H), 4.26 (m, 1H), 4.20 (m, 1H), 3.55 (m, 1H), 3.18 (m, 1H), 2.43 (m, 1H), 1.81 (s, 3H), 1.73 (m, 1H), 1.57 (dd, J=6.72, 1.83 HZ, 3H) MS: (M+H) =283, (M+23) =305, (M-H) =281, (2M-H) =563 -223- WO 99/54299 PCT/US99/07945 Example 52 (±)-(2R, 3S,5R,1'R,2'S)-2-(1 -Acetamido-2-hydroxy-2-vinyl)propyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN" OtBu CH 3 OtBu N AcHN. N O tB u OH O0 4 'OH 52A (±)-(2R,3S,5R,1'R,2'S) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxvcarbonyl-2 (1-acetamido-2-hydroxy-3-vinyl)propyl-3-(cis-propen-1-yvl)-pyrrolid idine-5-carboxyl ic Acid t-Butyl Ester. The title compounds were prepared according to the method described in Example 41B, substituting allyl magnesium bromide for ethyl magnesium bromide to provide (±)-(2R,3S,5R, 1'R,2'S)-l-t-butoxycarbonyl-2-(1-acetamido-2-hydroxy 2-vinyl)propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 2.0 mg, 5%) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 9.0 mg, 22%). (±)-(2R,3S,5R,1'R,2'S) MS: (M+H) = 453; (M-H) = 451. (±)-(2R,3S,5R,1'R,2'R) 1 H NMR (DMSO-d 6 ) 8 7.70 (d, J=9.3Hz, 1H), 5.80 (m, 1H), 5.51 (m, 1H), 5.30 (m, 1H), 5.00 (m, 2H), 4.58 (brd, 1H), 3.93 (m, 2H), 3.50 (m, 1H), 3.22 (br t, 1H), 2.02 (m, 3H), 1.88 (s, 3H), 1.56 (m, 4H), 1.41 (s, 9H), 1.36 (s, 9H) MS: (M-H)- = 451; (M+H)* = 452, (M+Na) = 475. -224- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH 'N HH OH 0 TFA 52B (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-3-vinyl)propyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-vinyl)propyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 2 mg, 100%). 1 H NMR (DMSO-d 6 ) 8 7.85 (d, J=9.3Hz, 1H), 5.81 (m, 1H), 5.42 (m, 1H), 5.28 (t, J=7.3Hz, 1H), 5.01 (br d, 2H), 3.99 (m, 2H), 3.57 (m, 2H), 3.08 (m, 1H), 2.33 (m, 1H), 2.26 (m, 1H), 2.07 (m, 1H), 1.81 (s, 3H), 1.57 (dd, J=1.4, 5.4Hz, 4H) MS: (M-H)- = 295; (M+H) = 297, (M+Na) = 319. -225- WO 99/54299 PCT/US99/07945 Example 53
CH
3 AcHN, /OH N 0f OH O TFA (±I)-(2R.,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-3-vinyl)propyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, I 'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-vinyl)propyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 6 mg, 100%). 1 H NMR (DMSO-d 6 ) 6 7.68 (d, J=9.2Hz, 1H), 5.78 (m, 1H), 5.48 (m, 1H), 5.24 (t, J=7.8Hz, 1H), 5.04 (m, 2H), 4.38 (t, J=7.0, 1H), 4.09 (t, J=7.0, 1H), 3.81 (t, J=4.7, 1H), 3.53 (t, J=8.5, 1H), 3.16 (m, 1H), 2.40 (m, 1H), 2.11 (m, 2H), 1.83 (s, 3H), 1.70 (m, 1H), 1.55 (dd, J=5.4, 1.4Hz, 3H) MS: (M-H) = 295; (M+H) = 297, (M+Na)* = 319. -226- WO 99/54299 PCT/US99/07945 Example 54 (±)-(2R, 3S, 5R, 1 'R,2'S)-2-(1-Acetamido-2-hydroxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. CH3 CH3 AcHN 1BHN OtBu AcHN. OtBu H B o oo O H 54A (±)-(2R,3S,5R,1'R,2'S) and (±)-(2R,3S,5R,1'R,2'R)-1-t-Butoxycarbonyl-2 (1-acetamido-2-hydroxy)pentyl-3-(cis-propen-l-yl)-pyrrolidine-5-carboxylic Acid t Butyl Ester. The title compounds were prepared according to the method described in Example 41B, substituting propyl magnesium bromide for ethyl magnesium bromide to provide (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 -acetamido-2 hydroxy)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 1 mg, 1%) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 32 mg, 39%). (±)-(2R,3S,5R,1'R,2'S) 1 HNMR (CDCI 3 ) 8 7.51 (d, J=8.2Hz, 1H), 5.46 (m, 2H), 4.17 (dd, J=3.1, 6.8Hz, 1H), 4.05 (m, 1H) 3.81 (t, J=3.4Hz, 1H), 3.54 (m, 1H), 3.21 (m, 1H), 2.60 (m, 1H), 2.02 (s, 3H), 1.70 (dt, J=3.0, 7.4Hz, 1H), 1.61 (d, J=5.4Hz, 3H), 1.54 (m, 1H), 1.47 (s, 9H), 1.44 (s, 9H), 1.32 (m, 4H), 0.90 (t, J=7.1 Hz, 3H) MS: (M+H) + = 455, (M+Na) + = 477; (M-H)- = 453. (±)-(2R,3S,5R,1'R,2'R) 1 H NMR (CDCI 3 ) 6 5.98 (d, J=9.5Hz, 1H), 5.60 (t, J=9.8Hz, 1H), 5.36 (m, 1H), 4.16 (m, 1H), 3.75 (d, J=10.1 Hz, 1H), 3.64 (m, 1H), 3.51 (m, 1H), 3.09 (brt, 1H), 2.68 (m, 1H), 2.02 (s, 3H), 1.81 (d, J=13.9Hz, -227- WO 99/54299 PCT/US99/07945 1H), 1.57 (m, 4H), 1.54 (dd, J=1.7, 5.1Hz, 3H), 1.46 (s, 9H), 1.45 (s, 9H), 0.88 (t, J=6.8Hz, 3H) MS: (M-H) = 453; (M+H) = 455.
CH
3 AcHN. OH N HH OH O TFA 54B (+)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)pentyl-3-(cis-propen-1 vil)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 1 mg, 100%). 1 H NMR (DMSO-d 6 ) 8 7.83 (d, J=9.2Hz, 1H), 5.43 (m, 1H), 5.23 (m, 1H), 3.98 (m, 1H), 3.56 (br t, 1H), 3.46 (m, 1H), 3.08 (m, 2H), 2.32 (m, 1H), 1.80 (s, 3H), 1.57 (dd, J=1.4, 5.4Hz, 4H), 1.43 (m, 2H), 1.23 (m, 2H), 0.85 (br t, 3H) MS: (M+H)* = 299, (M+Na) = 321. -228- WO 99/54299 PCT/US99/07945 Example 55
CH
3 AcHN. .. OH OH TFA (±)-(2R,3S. 5R,1'R,2'R)-2-(1 -Acetamido-2-hydroxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-l -t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0190 g, 100%). 1 H NMR (DMSO-d 6 ) 6 7.64 (d, J=9.3Hz, 1H), 5.48 (m, 1H), 5.24 (m, 1H), 4.38 (t, J=8.8Hz, 1H), 4.06 (m, 1H), 3.75 (m, 1H), 3.53 (t, J=8.5Hz, 1H), 3.16 (quint., J=8.5Hz, 1H), 2.41 (dt, J=12.8,7.3Hz, 1H), 1.82 (s, 3H), 1.70 (dt, 12.8,9.9Hz, 1H), 1.55 (dd, J=7.0,1.6Hz, 3H), 1.35 (m, 2H), 1.26 (m 2H), 0.86 (t, J=6.7Hz, 3H). MS: (M+H)* = 299, (M+Na) = 321, (M-H) = 297. -229- WO 99/54299 PCT/US99/07945 Example 56 (±)-(2R, 3S, 5R, 1 'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-methyl)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3
CH
3 AcHN. OtBu AcHN. OtBu HBoc O Boc 0 OH OH 56A (±)-(2R.3S,5R,1'R,2'S) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2 (1-acetamido-2-hydroxy-3-methyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic Acid t-Butyl Ester. The title compounds were prepared according to the method described in Example 41B, substituting isopropyl magnesium bromide for ethyl magnesium bromide to provide (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 -acetamido-2 hydroxy-3-methyl)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0092 g, 10%) and (±)-(2R,3S,5R,1'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-methyl)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0385 g, 40%). (±)-(2R,3S,5R,1'R,2'S) MS: (M+H) = 455, (M+Na) += 477, (2M+Na) =931, (M-H)-=453. (±)-(2R,3S,5R,1'R,2'R) MS: (M+H) = 455, (M+Na) = 477, (2M+Na) = 931, (M-H)-= 453. -230- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH 'N H 0 OH TFA 56B (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-3-methyl)butyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-methyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.010 g, 100%). 1 H NMR (DMSO-d 6 ) d 7.63 (d, J=9.2Hz, 1H), 5.48 (m, 1H), 5.23 (m, 1H), 4.44 (m, 1H), 4.24 (m, 1H), 3.57 (t, J=8.7Hz, 1H), 3.33 (dd, J=8.5,2.5Hz, 1H), 3.21 (quint., J=9.1Hz, 1H), 2.43 (dt, J=12.8,7.6Hz, 1H), 1.81 (s, 3H), 1.73 (dt, J=12.8,10.4Hz, 1H), 1.56 (dd, J=6.7,1.9Hz, 3H), 1.55 (m, 1H), 0.94 (d, J=6.7Hz, 3H), 0.78 (d, J=6.7Hz, 3H). MS: (M+H) + = 299, (M+Na) = 321, (M-H)- = 297, (M+CF 3 COOH)- = 411, (2M-H) = 595. -231- WO 99/54299 PCT/US99/07945 Example 57
CH
3 AcHN. HN'OH OH 0 OH TFA (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-3-methyl)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-methyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0433 g, 100%). 'H NMR (DMSO-d 6 ) d 7.88 (d, J=9.2Hz, 1H), 5.46 (m, 1H), 5.29 (m, 1H), 4.26 (t, J=8.5Hz, 1H), 4.11 (m, 1H), 3.67 (m, 1H), 3.39 (dd, J=9.8,1.8Hz, 1H), 3.15 (quint., J=9.1Hz, 1H), 2.42 (dt, J=12.8,7.9Hz, 1H), 1.81 (s, 3H), 1.73 (m, 1H), 1.62 (m, 1H), 1.57 (dd, J=7.0,1.6Hz, 3H), 0.88 (d, J=6.7Hz, 3H), 0.75 (d, J=6.7Hz, 3H). MS: (M+H) = 299, (M+Na) = 321, (M-H 2 0)* = 281, (M-H) = 297,
(M+CF
3 COOH)- = 411, (2M-H)- = 595. -232- WO 99/54299 PCT/US99/07945 Example 58 (±)-(2R,3S,5 R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy)hexyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 OtBu CH 3 OtBu AcHN.B AcHN- t. BOC'O Boc 0 OH OH 58A (±)-(2R,3S,5R,1'R,2'S) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2 (1-acetamido-2-hydroxy)hexyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t Butyl Ester. The title compounds were prepared according to the method described in Example 41B, substituting butyl magnesium bromide for ethyl magnesium bromide to provide (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)hexyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 2.0 mg, 8%) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)hexyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 6.0 mg, 24%).
CH
3 AcHN" "/OH OH OH 0 TFA 58B (±)-(2R, 3S,5R, 1'R.2'S)-2-(1-Acetamido-2-hydroxy)hexyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)hexyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t -233- WO 99/54299 PCT/US99/07945 butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 2.0 mg, 100%). 1 H NMR (DMSO-d 6 ) 6 8.34 (d, J=9.3Hz, 1H), 5.24 (m, 1H), 5.12 (m, 1H), 3.90 (m, 1H), 3.78 (m, 1H), 3.23 (m, 1 H), 2.90 (m, 1H), 2.14 (m, 1H), 1.80 (m, 1H), 1.75 (s, 3H), 1.52 (m, 3H), 1.45 (m, 1H), 1.08 (br s, 6H), 0.83 (br t, 3H). MS: (M-H)- = 311; (M+H) = 313. Example 59 AcHN. OH "OH O 'OH 0 TFA (±)-(2R,3S,5R,1 'R,2'R)-2-(1-Acetamido-2-hydroxy)hexyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)hexyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 6.0 mg, 100%). 1 H NMR (DMSO-d 6 ) 8 7.60 (d, J=9.3 Hz, 1H), 5.46 (m, 1H), 5.24 (t, J=9.2 Hz, 1H), 4.21 (t, J=8.3 Hz, 1H), 4.02 (t, J=7.9Hz, 1H), 3.74 (m, 1H), 3.47 (t, J=8.8, 1H), 3.12 (m, 1H), 2.37 (m, 1H), 1.81 (s, 3H), 1.64 (m, 1H), 1.55 (dd, J=1.5, 5.4 Hz, 3H), 1.29 (m, 6H), 0.86 (t, J=6.9, 3H) -234- WO 99/54299 PCT/US99/07945 MS: (M-H) = 311; (M+H) + = 313, (M+Na) + = 335. Example 60 (±)-(2R, 3S, 5R, 1 'R,2'S)-2-(1 -Acetamido-2-hydroxy-4-methyl)pentyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN. OtBu N OH Boc O OH O 60A (±)-(2R,3S,5R,1 'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy-4 methyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compounds were prepared according to the method described in Example 41B, substituting isobutyl magnesium bromide for ethyl magnesium bromide to provide (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy-4-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 31 mg, 51%). (±)-(2R,3S,5R,1'R,2'R) 1 H NMR (CDCl 3 ) 8 5.98 (d, J=9.5Hz, 1H), 5.61 (t, J=8.2Hz, 1H), 5.35 (m, 1H), 4.51 (dd, J=1.3, 3.1Hz, 1H), 4.15 (m, 1H), 3.74 (d, J=10.5Hz, 1H), 3.61 (m, 2H), 3.09 (t, J=7.5Hz, 1H), 2.71 (m, 1H), 2.02 (s, 3H), 1.81 (d, J=13.9Hz, 1H), 1.58 (brs, 1H), 1.54 (dd, J=1.7, 5.1Hz, 3H), 1.47 (s, 9H), 1.45 (s, 9H), 1.42 (m, 1H), 0.87 (dd, J=2.4, 6.7Hz, 6H) MS: (M-H)- = 467; (M+H)* = 469, (M+Na)* = 491. -235- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OtBu H BocO 0 60B (±)-(2R, 3S,5R,1'R)-l-t-Butoxycarbonyl-2-(1-acetamido-4-methyl-2 oxo)pentylI-3-(cis-propen-1 -vyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1'R,2'R)-1 -t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy-4 methyl)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (8.0 mg, 0.02 mmol) was reacted with Dess-Martin Periodinane (10 mg, 0.03 mmol) in dichloromethane (0.1 mL) at room temperature for 1 hour. The reaction was quenched with 1M aqueous sodium thiosulfate 1 mL and stirred for 20 minutes. The reaction was extracted with dichloromethane (3 x 1 mL). The organic layer was dried over magnesium sulfate,filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1:1: ethyl acetate/hexane to provide the title compound as a colorless semi-solid (yield: 4.8 mg, 61%).
CH
3 AcHN. NOtBu HBoc O OH 60C (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxvcarbonyl-2-(1-acetamido-2-hydroxy-4 methyl)pentyl-3-(cis-propen-1 -ylv)-pyrrolidine-5-carboxylic Acid t-Butvl Ester. (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-4-methyl-2 oxo)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (4.8 mg, 0.01 mmol) in methanol (0.1 mL) was reacted with sodium borohydride (2.0 mg, -236- WO 99/54299 PCT/US99/07945 0.05 mmol) at room temperature for 0.5 hours. The solvent was removed in vacuo and water (1 mL) was added. The aqueous layer was extracted with dichloromethane (3 x 1 mL). This organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1:1 ethyl acetate/hexane to provide the title compound (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2-hydroxy 4-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 2.4 mg, 51%).
CH
3 AcHN. OH 'N H 0 OH TFA 60B (±)-(2R,3S,5R,1 'R,2'S)-2-(1-Acetamido-2-hydroxy-4-methyl)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-( 1 acetamido-2-hydroxy-4-methyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 4.4 mg, 100%). 1 H NMR (D 2 0)5 5.45 (m, 1H), 5.15 (t, J=11.0Hz, 1H), 3.88 (m, 1H), 3.62 (t, J=8.0Hz, 1H), 3.43 (br t, 1H), 2.98 (m, 1H), 2.36 (m, 1H), 1.81 (s, 3H), 1.60 (m, 1H), 1.51 (m, 1H), 1.45 (dd, J=1.3, 5.4Hz, 3H), 1.17 (m, 3H), 0.74 (dd, J=6.7, 14Hz, 6H) MS: (M-H)- = 311; (M+H) = 313, (M+Na) = 335. -237- WO 99/54299 PCT/US99/07945 Example 61
CH
3 AcHN. KOH ' N HH O OH TFA (±)-(2R,3S,5R, 1 'R,2'R)-2-(1 -Acetamido-2-hydroxy-4-methyl)pentvl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1 -t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-4-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 1.7 mg, 85%). 1 H NMR (DMSO-d 6 ) 8 7.61 (d, J=9.8Hz, 1H), 5.45 (m, 1H), 5.24 (t, J=7.4Hz, 1H), 4.29 (br t, 1H), 4.0 (br t, 1H), 3.83 (m, 1H), 3.49 (t, J=8.8Hz, 1H), 3.13 (m, 1H), 2.39 (m, 1H), 1.82 (s, 3H), 1.68 (m, 2H), 1.55 (dd, J=1.4, 5.4Hz, 3H), 1.31 (m, 1H), 1.04 (m, 1H), 0.86 (dd, J=6.4, 8.3Hz, 6H) MS: (M-H)- = 311; (M+H) + = 313, (M+Na) = 335. -238- WO 99/54299 PCT/US99/07945 Example 62 (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamido-2-hydroxy)pent-3-vnyl)-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 CH 3 AcHN. <O N tBu AcHN. .' OtBu H Boc O H Boc " OH 'OH
CH
3
CH
3 62A (±)-(2R,3S,5R,1'R,2'S) and (±)-(2R,S,5R,1'R,2'R)-1l-t-Butoxvcarbonyl-2-(1 Acetamido-2-hydroxy)pent-3-ynyl)-3-(cis-propen- 1 -ylv)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compounds were prepared according to the method described in Example 41B, substituting propyn-1 -yl zinc for ethyl magnesium bromide to provide (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)pent-3-ynyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0073 g, 16%) and (±)-(2R,3S,5R, 1 'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)pent-3-ynyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0349 g, 77%). (±)-(2R,3S,5R,1'R,2'S) MS: (M+H)+=451, (M+Na)*=473, (2M+Na) =923,.(M-H)-=449. (±)-(2R,3S,5R,1'R,2'R) MS: (M+H) = 451, (M+Na)*=473, (2M+Na) =923, (M-H)=449. -239- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH H O OH
OH
3 TFA 62B (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)pent-3-ynyl)-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)pent-3-ynyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0052 g, 100%). 1 H NMR (DMSO-d 6 ) d 7.97 (d, J=8.3 Hz, 1H), 5.48 (m, 1H), 5.25 (m, 1H), 4.35-4.20 (m, 3H), 3.67 (m, 1H), 3.18 (quint., 8.8Hz, 1H), 2.41 (dt, J=12.7,7.8Hz, 1H), 1.84 (s, 3H), 1.81 (d, J=1.9Hz, 3H), 1.63 (m, 1H), 1.59 (dd, J=6.9,2.0Hz, 3H). MS: (M+H) = 295, (M+Na) = 317, (M-H)- = 293, (M+CF 3 COO-)-=407, (2M H)= 587. -240- WO 99/54299 PCT/US99/07945 Example 63
CH
3 AcHN. OH SII HH O OH
CH
3 TFA (±)-(2R,3S,5R, 1 'R,2'R)-2-(1-Acetamido-2-hydroxy)pent-3-ynyl) -3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)pent-3-ynyl-3-(cis-propen- -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0540 g, 100%). 1 H NMR (DMSO-d 6 ) d 7.90 (d, J=8.8 Hz, 1H), 5.50 (m, 1H), 5.25 (m, 1H), 4.40-4.35 (m, 2H), 4.28 (m, 1H), 3.71 (t, J=8.0 Hz, 1H), 3.18 (quint., 8.3Hz, 1H), 2.42 (dt, J=13.2,7.4Hz, 1H), 1.87 (s, 3H), 1.82 (d, J=1.9Hz, 3H), 1.71 (dt, J=12.7,10.0Hz, 1H), 1.57 (dd, J=6.9,1.5Hz, 3H). MS: (M+H) = 295, (M+Na) = 317, (M-H)- = 293, (M+CF 3 COO-)=407. Example 64 -241- WO 99/54299 PCT/US99/07945 (±)-(2R, 3S, 5R, 1'R,2'R)-2-(1 -Acetamido-2-hydroxy-2-heptafluoropropyl)ethyl-3 (cis-propen-1-vl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OH Hjn oc 0
CF
3
CF
2
CF
2 'OH 64A (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy-2 heptafluoropropyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butvl Ester. (±)-(2R,3S,5R, 1 'R)-1 -t-Butoxycarbonyl 2-(1-acetamido-1 -formyl)methyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (41 mg, 0.10 mmol) and heptafluoropropyl iodide (0.144 mL, 1.0 mmol, 10 equivalents) in THF (2 mL) were reacted with 1M phenylmagnesium bromide (0.90 mL, 0.90 mmol, 9 equivalents) at -78 oC for 5 minutes. The reaction mixture was allowed to warm to room temperature over 1 h. The reaction was quenched with saturated aqueous ammonium chloride (10 mL) and water (10 mL) followed by extraction using ethyl acetate (3 X 25 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/2: ethyl acetate/hexane to provide the title compound (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy 2-heptafluoropropyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 12.6 mg, 22%). (±)-(2R,3S,5R,1'R,2'R) MS: (M+H)+=581, (M+Na)+=603, (2M+Na)*=1 183, (M-H)=579. -242- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH H 0
CF
3
CF
2
CF
2 'OH TFA 64B (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-2 heptafluoropropyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-heptafluoropropyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 0.003 g, 100%). 1 H NMR (DMSO-d 6 ) d 7.84 (d, J=9.3 Hz, 1H), 5.45 (m, 1H), 5.26 (m, 1H), 4.71 (t, J=9.7 Hz, 1H), 4.63 (d, J=22.0 Hz, 1H), 4.51 (m, 1H), 3.59 (t, J=9.3 Hz, 1H), 3.19 (quint., 8.3Hz, 1H), 2.43 (dt, J=12.7,7.3Hz, 1H), 1.76 (s, 3H), 1.74 (m, 1H), 1.53 (dd, J=6.8,1.4Hz, 3H). MS: (M+H) = 425, (M+Na) = 447, (M-H)- = 423, (2M-1)- = 847. Example 65 -243- WO 99/54299 PCT/US99/07945 (±)-(2R,3S, 5R, 1'R,2'S)-2-(1 -Acetamido-2,4-dihydroxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OtBu HN ' oc O OH OH 65A (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2,4 dihydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy-3 ethoxycarbonyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (35 mg, 0.07 mmol) was reacted with lithium borohydride (8 mg, 0.35 mmol) in THF (5 mL) at 250C and reacted for 3 hours. The reaction was quenched with saturated aqueous ammonium chloride (2 mL) and water (2 mL) followed by extraction using dichloromethane (2 X 10 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 5% methanol in dichloromethane to provide the title compound (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl 2-(1 acetamido-2,4-dihydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 14 mg, 44%). (±)-(2R,3S,5R,1'R,2'S) =MS: (M+H)*=457, (M-H)- =455 -244- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH ,'N K HH0 OH OHTFA OH 65B (±)-(2R.3S,5R,1'R,2'S)-2-(1-Acetamido-2,4-dihydroxy)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'S)-1 -t-butoxycarbonyl-2-(1 acetamido-2,4-dihydroxy)butyl 3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. 'H NMR (DMSO-d 6 ) 5 7.93 (d, J=9.0Hz, 1H), 5.56(m, 1H), 5.31(m, 1H), 4.43 (m, 1H), 4.14 (m, 1H), 3.69 (m, 1H), 3.63 (m, 1H), 3.23 (m, 2H), 3.07 (m, 1H), 2.43 (m, 1H), 2.06 (s, 3H), 1.83 (m, 2H), 1.79 (m, 1H), 1.62 (dd, J=6.71, 1.22 Hz, 3H) MS: (M+H)+=301, (M-H) =299. Example 66 -245- WO 99/54299 PCT/US99/07945 (±)-(2R,3S, 5R, 1'R,2'R)-2-(1 -Acetamido-2,4-dihydroxy) b utyl-3-(cis-p ropen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OtBu N -K H BocO OH OH 66A (2R,3S.5R,1 'R,2'R)-1l-t-Butoxvcarbonyl-2-(1-acetamido-2,4 dihydroxy)butvl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 65A substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-ethoxycarbonyl)ethyl-3-(cis-propen- 1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy-3-ethoxycarbonyi)ethyl-3-(cis-propen-1-yl)-pyrrol Idine 5-carboxylic acid t-butyl ester. (yield: 11 mg, 70%). 1 H NMR (CDCI 3 ) 6 5.58(m, 1H), 5.38(m, 1H),4.16(m, 1H), 4.05(m, 1H), 3.97(m, 1H), 3.78(m, 2H), 3.20(m, 1H), 2.66(m, 1H) 2.54(m, 1H), 2.04(s, 3H), 1.80(m, 1H), 1.55(m, 2H), 1.47(s, 9H), 1.44(s, 9H) MS: (M+H)+=457, (M-H) =455 -246- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH H 0 OH TFA OH 66B (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2,4-dihydroxy)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2,4-dihydroxy)butyl 3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 8 mg, 96%). 'H NMR (DMSO-d 6 ) 5 7.91 (d, J=9.1Hz, 1H), 5.50 (m, 1H), 5.25 (m, 1H), 4.43 (m, 1H), 4.30 (m, 1H), 4.22 (m, 1 H), 3.94 (m, 1 H), 3.86 (m, 1H), 3.62 (m, 1H), 3.18 (m, 1H), 2.43 (m, 1H), 1.85 (s, 3H), 1.75 (m, 1H), 1.65 (m, 2H), 1.58 (dd, J=6.70, 1.81 Hz, 3H). MS: (M+H)* =301, (M-H) =299. -247- WO 99/54299 PCT/US99/07945 Example 67 (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-2-(phenylacetylen-1-yl))ethyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3
CH
3 HBoc O H C Bo0 'OH < OH Ph Ph 67A (±)-(2R,3S,5R,1'R,2'R) and (±)-(2R,3S,5R,1'R,2'S) -1-t-Butoxycarbonyl-2 (1-acetamido-2-hydroxy-2-(phenylacetylen-1 -yl))ethyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compounds were prepared according to the method described in Example 41B, substituting lithium phenylacetylide for ethyl magnesium bromide to provide (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2-hydroxy 2-(phenylacetylen-1-yl))ethyl -3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 0.0010 g, 4%) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl 2-(1 -acetamido-2-hydroxy-2-(phenylacetylen-1-yl))ethyl -3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0050 g, 21%). (±)-(2R,3S,5R,1'R,2'S) MS: (M+H) =513, (M+Na)+=535, (2M+Na) =1047, (M-H)=511. (±)-(2R,3S,5R,1'R,2'R) MS: (M+H) =513, (M+Na) =535, (2M+Na)+=1047, (M-H)-=511. -248- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH HH 0 h OH TFA Ph 67B (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-2-(phenylacetvlen-l yl))ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound is prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-(phenylacetylen-1-yl))ethyl-3-(cis-propen-1-yl)-pyrrolidine 5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester. Example 68 (±)-(2R,3S,5R, 1 'R,2'R)-2-(1 -Acetamido-2-hydroxy-2-(phenylacetylen-1 -yl))ethyl 3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 N AcHN N" OH H " OH <-'OH Ph TFA 68A (±)-(2R,3S,5R.,1 'R,2'R)-2-(l1-Acetamido-2-hydroxy-2-(phenylacetylen- 1 yl))ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1 'R,2'R)-1 -t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-(phenylacetylen-1-yl))ethyl-3-(cis-propen-1-yl)-pyrrolidine -249- WO 99/54299 PCT/US99/07945 5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yi)-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 0.0034 g, 100%). 1 H NMR (DMSO-d 6 ) 6 9.2 (bs, 1H), 8.04 (d, J=9.2 Hz, 1H), 7.45-7.35 (m, 5H), 5.50 (m, 1H), 5.29 (m, 1H), 4.64 (d, J=4.9, 1H), 4.5-4.4 (m, 2H), 3.81 (m, 1H), 3.22 (quint., J=8.5Hz, 1H), 2.45 (dt, J=12.8,7.3Hz, 1H), 1.89 (s, 3H), 1.74 (dt, J=12.7, 10.0Hz, 1H), 1.58 (dd, J=7.3,1.8Hz, 3H). MS: (M+H) = 357, (M+Na) = 379, (M-H) = 355. Example 69 (±)-(2R,3S,5R. 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-ethyl)pentyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OtBu ~Nf H Boc O 69A (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-oxo-3 ethyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxvlic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 42A, substituting (±)-(2R,3S,5R,1'R,2',R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-ethyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 8 mg, 81%). MS: (M+H)+=481, (M-H) =479 -250- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OtBu H Boc O OH 69B (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-hydroxy-3 ethyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-ButyI Ester. The title compound was prepared according to the method described in Example 42B, substituting (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1-acetamido 2-oxo-3-ethyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (2R,3S,5R,1'R)-1l-t-butoxycarbonyl 2-(1-acetamido-2-oxo)butyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 5 mg, 63%). MS: (M+H)+=483, (M-H)- =481
CH
3 AcHN. OH N i CHH O OH TFA 69C (±)-(2R,3S.5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-3-ethyl)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-ethyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 4 mg, 95%). -251- WO 99/54299 PCT/US99/07945 1 H NMR (DMSO-d 6 ) 8 7.67(d, J=8.9Hz, 1H), 5.48(m, 1H), 5.23(m, 1H), 4.42(m, 1H), 4.21(m, 1H), 3.58(m, 2H), 3.22(m, 1H), 2.43(m, 1H), 1.82(s, 3H), 1.74(m, 1H), 1.58(dd, J=6.71, 1.23 Hz, 3H), 1.52(m, 1H), 1.38(m, 1H), 1.29(m, 2Hz), 1.13(m, 1H), 0.80(m, 6H) MS: (M+H) =327, (M-H)-=325 Example 70 (±)-(2R,3S. 5R, 1 'R,2'R)-2-(1 -Acetamido-2-hydroxy-3-ethyl)pentyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 o AcHN. OtBu H BOC O 'OH 70A (±)-(2R, 3S,5R, I'R,2'R)-1l-t-Butoxvcarbonvl-2-(l-Acetamido-2-hvdroxv-3 ethyl)penyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic Acid t-ButvI Ester. The title compound was prepared according to the method described in Example 41B, substituting 3-pentyl magnesium bromide in place of ethyl magnesium bromide (yield: 13mg, 45%). MS: (M+H)+=483, (M-H)- =481 -252- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH N 0 OH TFA 70B (±)-(2R, 3S.5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-3-ethyl)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-ethyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 3 mg, 96%). 1 H NMR (DMSO-d 6 ).6 7.85 (d, J=9.2 Hz, IH), 5.47 (m, 1H), 5.30 (m, 1H), 4.28 (m, 1H), 4.19 (m, 1H), 3.67 (m, 1H), 3.58 (m, 1H), 3.17 (m, 1H), 2.43 (m, 1H), 1.81 (s, 3H), 1.63 (m, 1H), 1.58 (dd, J=6.71, 1.82 Hz, 3H), 1.40 (m, 2H), 1.28 (m, 1H), 1.10 (m, 1H), 1.05 (m, 1H), 0.83 (m, 6H) MS: (M+H)+=327, (M-H)-=325 -253- WO 99/54299 PCT/US99/07945 Example 71 (±)-(2R,3S,5R, 1' R,2'R)-2-(1 -Acetamido-2-hydroxy-2-phenyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN., OtBu SN ' OH Boc6 'OH 71A (±)-(2R,3S,5R,1 'R,2'R)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-hydroxy-2 phenyl)ethyl-3-(cis-propen-1 -ylv)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 41 B, substituting phenyl magnesium bromide in place of ethyl magnesium bromide (yield: 36 mg, 60%). MS: (M+H) = 489, (M+Na)+= 511, (M-H)- = 487.
CH
3 AcHN.- N OH OH 0 OH TFA 71B (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-2-phenyl)ethyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-phenyl)ethyl-3-(cis-propen-1 -yl)-pyrrolid ine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 -254- WO 99/54299 PCT/US99/07945 acetamido-2-hydroxy)butyl-3-(cis-propen-l1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 5.5 mg, 100%). 'H NMR (DMSO-d 6 ) d 7.79 (d, J= 9.2Hz, 1H), 7.36 (m, 2H), 7.31 (m, 2H), 7.22 (m, 1H), 5.49 (m, 1H), 5.22 (m, 1H), 4.94 (d, J= 3.0Hz, 1H), 4.52 (m, 1H), 4.35 (m, 1H), 3.62 (t, J= 8.5Hz, 1H), 3.22 (m, 1H), 2.46 (m, 1H), 1.77 (m, 1H), 1.65 (s, 3H), 1.57 (dd, J= 6.7, 0.8Hz, 3H). MS: (M+H)+= 333, (M+Na)+ = 355, (M-H)- = 331. Example 72 (±)-(2R. 3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-2-phenyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt /=>
CH
3 AcHN., N OH H Boc 72A (±)-(2R.3S,5R,1'R)-l1-t-Butoxcarbonyl-2-(1-Acetamido-2-oxo-2 phenyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 42A, substituting (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-phenyl)ethyl-3-(cis-propen- l -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl 2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 24 mg, 84%). MS: (M+H)*= 487, (M+Na) = 509, (M-H)- = 485. -255- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH OH Boc0 \ O 0 OH 72B (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-hydroxy-2 phenyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 42B, substituting (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1-acetamido 2-oxo-2-phenyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (2R,3S,5R,1'R)-1l-t-butoxycarbonyl 2-(1-acetamido-2-oxo)butyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 7.9 mg, 52%). MS: (M+H) = 489, (M+Na)*= 520, (M-H)- = 487.
CH
3 AcHN, OH N H 0 OHTFA 72C (±)-(2R,3S,5R,1'R.2'S)-2-(1-Acetamido-2-hydroxy-2-phenyl)ethyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-phenyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 7.5 mg, 100%). -256- WO 99/54299 PCT/US99/07945 1 H NMR (DMSO-d 6 ) d 7.83 (d, J= 9.2Hz, 1H), 7.36 (m, 2H), 7.32 (m, 2H), 7.25 (m, 1H), 5.47 (m, 1H), 5.33 (m, 1H), 4.54 (d, J= 9.8Hz, 1H), 4.36 (m, 1H), 4.23 (m, 1H), 3.78 (m 1H), 3.20 (m, 1H), 2.43 (m, 1H), 1.63 (m, 1H), 1.56 (dd, J= 6.7, 1.2Hz, 3H), 1.53 (s, 3H). MS: (M+H)*= 333, (M+Na) = 355, (M-H)- = 331. Example 73 (±)-(2R,3S,5R, 1'R,2'R)-2-(1 -Acetamido-2-hydroxy-2-(thiophen-2-yl))ethyl-3-(cis propen-1-ylv)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. 0 Boc 0 OH S 73A (±)-(2R,3S,5R. 1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy-2 (thiophen-2-yl))ethyl-3-(cis-propen-1 -vyl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1 'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-2-formyl)ethyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (40 mg, 0.098 mmol) in THF (2 mL) was added dropwise to a solution of 2-thienyllithium (1M in THF, 0.505 mmol, 5 equivalents) in THF (1 mL) at 25 oC and reacted for 20 minutes. The reaction was quenched with saturated aqueous ammonium chloride (2 mL) and water (5 mL) followed by extraction using dichloromethane (2 X 10 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/1: ethyl acetate/hexane to provide the title compound (yield: 9.5 mg, 20%). MS: (M+H) = 495, (M+Na) = 517, (M-H)- = 493. -257- WO 99/54299 PCT/US99/07945
CH
3 AcHN. , OH H OH S TFA 73B (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-2-(thiophen-2-yl))ethyl 3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1 -t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-(thiophen-2-yl))ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 4.3 mg, 100%). 1 H NMR (DMSO-d 6 ) 8 7.86 (d, J= 9.8Hz, 1H), 7.63 (dd, J= 5.4, 1.0 Hz, 1H), 7.07 (m, 1H), 6.98 (m, 1H), 5.58 (m, 1H), 5.43 (m, 1H), 4.55 (m, 1H), 4.39 (m, 1H), 3.72 (m, 1H), 3.11 (m, 2H), 2.43 (m, 1H), 2.04 (s, 3H), 1.80 (m, 1H), 1.57 (m, 3H). MS: (M+H) = 339, (M+Na)+ = 361, (M-H)- = 337. Example 74 -258- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-(4-methylthiazol-2-yl))propyl 3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3
CH
3 o AcHN. N .OtBu AcHN. OtBu N (N Boc 0 O HB OH OH N S N S
H
3 C H 3 C 74A (±)-(2R,3S,5R,1'R,2'S) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2 (1-Acetamido-2-hydroxy-3-(4-methylthiazol-2-yl))propyl-3-(cis-propen- 1-yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. 1.6 M n-Butyllithium (0.125 mL, 0.20 mmol, 4 equivalents) was added to a solution of 2,4-dimethylthiazole (28.3 mg, 0.25 mmol, 5 equivalents) in 1 mL of THF at -78 'C and reacted for 30 minutes. ((±)-(2R,3S,5R,1'R)-1-t butoxycarbonyl 2-(1-acetamido-2-formyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester (20.5 mg, 0.050 mmol) in THF (1 mL) was added dropwise to the above solution and reacted for 30 minutes at -78 oC and then for 30 minutes at room temperature. The reaction mixture was quenched with saturated aqueous ammonium chloride (5 mL) and water (5 mL) followed by extraction using dichloromethane (3 X 25 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/2: ethyl acetate/hexane to provide (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2-hydroxy-3 (4-methylthiazol-2-yl))propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 3.3 mg, 13%) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy-3-(4-methylthiazol-2-yl))propyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic acid t-butyl ester (yield: 7.5 mg, 29%). -259- WO 99/54299 PCTIUS99/07945 (2R,3S,5R,1'R,2'S) MS: (M+H) =524, (M+Na) =546, (2M+Na) =1069, (M H)=522. (2R,3S,5R,1'R,2'R) MS: (M+H) =524, (M+Na) =546, (2M+Na)*=1069, (M H)-=522.
CH
3 AcHN. OH N HH OH N S TFA
H
3 C 74B (±)-(2R.,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-3-(4-methylthiazol-2 yl))pDropyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-(4-methylthiazol-2-yl))propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 0.0030 g, 100%). 1 H NMR (DMSO-d 6 ) 8 9.0 (bs, 1H), 8.10 (d, J=8.3Hz, 1H), 7.11 (d, J=1.0 Hz, 1H), 5.48 (m, 1H), 5.30 (m, 1H), 4.30 (m, 1H), 4.10 (m, 1H), 3.88 (dt, J=9.4,2.6Hz, 1H), 3.78 (m, 1H), 3.25-3.15 (m, 2H), 2.93 (dd, J=15.1,8.3Hz, 1H), 2.41 (dt, J=12.3,7.3Hz, 1H), 2.33 (d, J=1.0Hz, 3H), 1.86 (s, 3H), 1.66 (dt, J=12.7, 10.3Hz, 1H), 1.61 (dd, J=6.8,1.5Hz, 3H). MS: (M+H) = 368, (M+Na)+ = 390, (M-H)- = 366. -260- WO 99/54299 PCT/US99/07945 Example 75 (±)-(2R,3S.5R, 1'R.2'R)-2-(1 -Acetamido-2-hydroxy-3-(4-methylthiazol-2-yl))propyl 3-(cis-propen-1 -yil)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OH 'OH N'S TFA
H
3 C The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-(4-methylthiazol-2-yl))propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 0.0030 g, 100%). 1 H NMR (DMSO-d 6 ) d9.0 (bs, 1H), 7.77 (d, J=9.3Hz, 1H), 7.11 (s, 1H), 5.47 (m, 1H), 5.25 (m, 1H), 4.45 (m, 1H), 4.20 (m, 2H), 3.58 (t, J=9.1Hz, 1H), 3.19 (m, 1H), 2.96 (m, 2H), 2.41 (m, 1H), 2.33 (d, J=1.0Hz, 3H), 1.85 (s, 3H), 1.73 (dt, J=12.7, 10.3Hz, 1H), 1.54 (dd, J=6.9,1.5Hz, 3H). MS: (M+H) = 368, (M+Na)+ = 390, (M-H)- = 366, (M+CF 3 COOH)-=480, (2M-H)-=733. Example 76 -261- WO 99/54299 PCT/US99/07945 (±)-(2R,3S, 5R, 1'R,2'RS)-2-(1 -Acetamido-2-hydroxy-3-(thiazolin-2-vl))Dropyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OH 'N H Boc o OH N S \-j 76A (±)-(2R,3S,5R,1'R,2'RS)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2 hydroxy-3-(thiazolin-2-yl))propyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R, 1 'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-2-formyl)ethyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (20.5 mg, 0.05 mmol) in THF (1 mL) was added dropwise to a solution of the (thiazolin-2-yl)methyl lithium (0.20 mmol, 4 equivalents, prepared from 0.025 g of 2-methylthiazoline and 0.125 mL of 1.6 M n-BuLi at -78 oC) in THF (2 mL) at -78 OC and reacted for 30 minutes. The reaction was quenched with saturated aqueous ammonium chloride (5 mL) and water (5 mL) followed by extraction using dichloromethane (3 X 20 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/1: ethyl acetate/hexane to provide the title compound as a mixture of isomers (yield: 10 mg, 40%). MS: (M+H)*= 512, (M+Na)+=534, (M-H)-=510. -262- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH 'N H 0 OH N S TFA 76B (±)-(2R,3S,5R,1'R,2'RS)-2-(1-Acetamido-2-hydroxy-3-(thiazolin-2 yl))propyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compounds were prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'RS)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-(thiazolin-2-yl))propyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester n place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 0.003 g, 100%). Major isomer 1 H NMR (DMSO-d 6 ) 6 8.88 (m, 1H), 7.76 (d, J=8.8Hz, 1H), 5.46 (m, 1H), 5.19 (m, 1H), 4.69 (m, 1H), 3.90 (m, 1H), 3.85 (m, 1H), 3.49 (m, 2H), 3.35 (t, J=9.0Hz, 1H), 3.29 (dd, J=17.6,5.9Hz, 1H), 3.04 (t, J=8.9Hz, 1H), 2.78 (dd, J=17.6,8.1Hz, 1H), 2.7-2.55 (m, 2H), 1.75 (s, 3H), 1.70 (m, 1H), 1.56 (dd, J=6.8,1.5Hz, 3H). MS: (M+H) = 356, (M+Na)*=378, (2M+Na) =733, (M-H)-=354. Example 77 -263- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R. 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-3,3-difluoro-3-vinyl)propyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OtBu N H Boc OH FF 77A (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-hydroxv-3,3 difluoro-3-vinyl)propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-2-formyl)ethyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (41 mg, 0.10 mmol) and 1,1-difluoroallyl iodide (94 mg, 0.60 mmol, 6 equivalents) in THF (2 mL) was reacted with zinc dust (33 mg, 0.50 mmol, 5 equivalents) at 0 0 C for 5 minutes and then at room temperature for 4 hours. The reaction mixture was quenched with saturated aqueous ammonium chloride (15 mL) and water (15 mL) and extracted with 3 X 25 mL dichloromethane. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/3: ethyl acetate/hexane to provide the title compound (yield: 35 mg, 71%). MS: (M+H)+=489, (M+Na)*=511, (2M+Na) =999, (M-H)-=487. -264- WO 99/54299 PCT/US99/07945
CH
3 AcHN. N OH 'N HH 0 OH F F TFA 77B (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-3,3-difluoro-3 vinvyl)Dropyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'S)-l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3,3-difluoro-3-vinyl)propyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 0.0026 g, 96%). 1 H NMR (DMSO-de) d7.68 (d, J=7.8Hz, 1H), 5.97 (m, 1H), 5.55-5.45 (m, 2H), 5.43 (m, 1H), 5.23 (m, 1H), 4.45 (m, 2H), 4.10 (m, 1H), 3.16(quint. J=9.1Hz, 1H), 2.41 (dt, J=12.8,7.3Hz, 1H), 1.72 (s, 3H), 1.70 (dt, J=12.8, 10.3Hz, 1H), 1.61 (dd, J=6.7,1.2Hz, 3H). MS: (M+H)*=333, (M+Na) =355, (M-H)-= 331, (2M-H)-=663. Example 78 -265- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R, 1'R,2'R)-2-( 1 -Acetamido-2-hydroxy-3,3-difluoro-3-vinyl)propyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OtBu HN ' SBoc O F F 78A (±)-(2R,3S,5R, 1 'R)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-oxo-3,3-difluoro 3-vinyl)propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 42A, substituting (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3,3-difluoro-3-vinyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (2R,3S,5R,1'R,2'R)-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester to provide (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1-acetamido-2-oxo-3,3-difluoro 3,3-difluoro-3-vinyl)propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0050g, 44%). MS: (M+H)*=487, (M+Na)+=509, (M-2F) =448, (M-H)-=485. -266- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OtBu H Boc OH FF 78B (±)-(2R.3S,5R, 1'R,2'R)-1-t-Butoxycarbonyl-2-(1-Acetamido-2-hydroxy-3,3 difluoro-3-vinyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 42B, substituting (±)-(2R,3S,5R,1'R)-1l-t-butoxycarbonyl-2-(1-acetamido 2-oxo-3-difluoro-3-vinyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (2R,3S,5R,1'R)-2-(1-acetamido-2-oxo)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester.
CH
3 AcHN. iOH
H
H OH F F TFA 78C (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-3,3-difluoro-3 vinyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid The title compound is prepared according to the method described in Example 41 C, substituting (+)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3,3-difluoro-3-vinyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester. Example 79 -267- WO 99/54299 PCT/US99/07945 (±)-(2R, 3S,5R, 1'R,2'R)-2-( 1 -Acetamido-2-hydroxy-2-(cis-buten-2-ylI))ethyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OtBu H r BocO
H
3 C 'OH
CH
3 79A (±)-(2R,3S,5R,1'R.2'R)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-hydroxy-2 (cis-buten-2-yl))ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-2-formyl)ethyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (30 mg, 0.073 mmol) in THF (5 mL) was reacted with cis-2-buten-2-yl lithium (0.75 mL (0.5M), 0.37 mmol) at 25 0 C for 45 min. The reaction was quenched with saturated aqueous ammonium chloride (5 mL) and water (5mL) followed by extraction using dichloromethane (2 X 10 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/1: ethyl acetate/hexane to provide the title compound (yield: 20 mg, 59%). 1 H NMR (CDCl 3 ) 5 6.19(d, J=8.9 Hz, 1H), 5.61(m, 1H), 5.35(m, 1H), 5.27(m, 1H), 4.48(m, 1H), 4.18(m, 1H), 4.77(m, 2H), 3.10(m, 1H), 2.72(m, 1H), 1.99(s, 3H), 1.82(m, 1H), 1.73(m, 3H), 1.55(m, 6H), 1.47(s, 9H), 1.44(s, 9H) MS: (M+H) = 467, (M-H)-= 465 -268- WO 99/54299 PCT/US99/07945 CH 3 AcHN. . N OH
H
3 C 'OH O
CH
3 TFA 79B (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-2-(cis-buten-2-yl))ethyl 3-(cis-propen-1 -ylv)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-(cis-buten-2-yl))ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 4 mg, 96%). 1 H NMR (DMSO-d 6 ) 5 8.09(d, J=9.0 Hz, 1H), 5.50(m, 1H), 5.32(m, 1H), 5.16(m, 1H), 4.50(m, 1H), 4.38(m, 1H), 4.19(m, 1H), 3.43(m, 1H), 3.20(m, 1H), 2.43(m, 1H), 1.88(s, 3H), 1.74(m, 1H), 1.70(s, 3H), 1.62(m, 3H), 1.58(m, 3H) MS: (M+H) =311, (M-H) =309 Example 80 -269- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R. 1'R,2'R.3'R) and (±)-(2R,3S,5R,1'R,2'R,3'S)-2-(1-Acetamido-2 hydroxy-3-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3
CH
3 AcHN. OtBu AcHN N OBu H Boc O H B
O
c O OH OH 80A (±)-(2R,3S,5R,1'R,2'R,3'R) and (±)-(2R,3S,5R,1'R,2'R,3'S)-1-t Butoxvcarbonyl-2-(1-acetamido-2-hydroxy-3-methyl)pentvl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1 'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-1 -formyl)methyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (60 mg, 0.15 mmol) in THF (1 mL) was added dropwise to a solution of 2-butylmagnesium bromide (3M in ether) (0.45 mL, 0.85 mmol) at room temperature and reacted for 40 minutes. The reaction was quenched with saturated NH 4 CI (1 mL) followed by extraction using dichloromethane (3 x 1 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/4: ethyl acetate/hexane to provide the title compounds (±)-(2R,3S,5R, 1 'R,2'R,3'S)-1l-t-butoxycarbonyl 2-(1-acetamido-2 hydroxy-3-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester Rf= 0.65 (1:1 ethyl acetate: hexanes) (yield: 19 mg, 27%) and (±) (2R,3S,5R,1'R,2'R,3'R)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy-3 methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester Rf= 0.5) (1:1 ethyl acetate: hexanes) (yield: 19 mg, 27%). Rf= 0.65 1 H NMR (CDCI 3 ) 8 5.98 (d, J=8.8Hz, 1H), 5.62 (t, J=10.5Hz, 1H), 5.35 (m, 1H), 4.66 (d, J=4.4Hz, 1H), 4.16 (d, J=9.5Hz, 1H), 3.78 (m, 3H), -270- WO 99/54299 PCT/US99/07945 3.12(m, 2H), 2.73 (m, 1H), 2.0 (s, 3H), 1.81 (d, J=13.2Hz, 1H), 1.54 (brs, 3H), 1.47 (s, 9H), 1.44 (s, 9H), 1.25 (m, 1H), 0.81 (m, 6H) MS: (M-H)- = 467; (M+H)* = 469. Rf= 0.5 1 H NMR (CDCI 3 ) 6 6.00 (d, J=10.2Hz, 1H), 5.61 (brt, 1H), 5.36 (m, 1H), 4.58 (d, J=4.7Hz, 1H), 4.14 (d, J=8.8Hz, 1H), 3.82 (m, 3H), 3.13 (m, 2H), 2.73 (m, 1H), 1.99 (s, 3H), 1.80 (d, J=13.9Hz, 1H), 1.54 (brs, 3H), 1.46 (s, 9H), 1.44 (s, 9H), 1.43 (m, 1H), 0.97 (d, J=6.8Hz, 3H), 0.81 (t, J=7.2Hz, 3H) MS: (M-H) = 467; (M+H) = 469.
CH
3 AcHN , OH N ? S -oHH ° OH TFA 80B (±)-(2R,3S,5R,1 'R,2'R,3'S)-2-(1-Acetamido-2-hydroxy-3-methyl)pentyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt (±)-(2R,3S,5R, 1'R,2'R,3'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy 3-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (2.5 mg, 0.005 mmol) was reacted with trifluoroacetic acid (0.8 mL) in dichloromethane (0.2 mL) at room temperature for 6 hrs. The reaction was concentrated in vacuo overnight and triturated with acetonitrile (2 x 1 mL) to provide the title compound (yield: 2.0 mg, 100%). 1H NMR (DMSO-d6) 8 7.68 (d, J=8.8Hz, 1H), 5.45 (m, 1H), 5.23 (t, J=7.3Hz, 1H), 4.24 (brt, 1H), 4.18 (m, 1H), 3.52 (t, J=7.3Hz, 1H), 3.45 (m, 1H), 3.16 (m, 1H), 2.38 (m, 1H), 1.83 (s, 3H), 1.68 (m, 1H), 1.58 (dd, J=2.0, 4.8Hz, 3H), 1.37 (m, 2H), 0.99 (m, 1H), 0.89 (d, J=6.8Hz, 3H), 0.79 (t, J=7.4Hz, 3H) -271- WO 99/54299 PCT/US99/07945 MS: (M-H)- = 311; (M+H)+ = 313, (M+Na)+ = 335. Example 81 (±)-(2R,3S,5R,1'R,2'R,3'R)-2-(1-Acetamido-2-hydroxy-3-methyl)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. H N H OOH H S TFA The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'R,3'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (Rf= 0.5, 1:1, ethyl acetate:hexanes) in place of (±) (2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 1.6 mg, 76%). 1 H NMR (DMSO-d 6 ) 8 7.55 (d, J=9.3Hz, 1H), 5.45 (m, 1H), 5.23 (m, 1H), 4.31 (brt, IH), 4.20 (t, J=8.3Hz, 1H), 3.51 (t, J=9.3Hz, 1H), 3.43 (d, J=7.4Hz, 1H), 3.17 (m, 1H), 2.40 (m, 1H), 1.80 (s, 3H), 1.70 (m, 1H), 1.55 (dd, J=1.4, 5.4Hz, 3H), 1.36 (m, 2H), 1.14 (m, 1H), 0.84 (t, J=7.3Hz, 3H), 0.73 (d, J=6.9Hz, 3H) MS: (M-H)- = 311; (M+H) + = 313, (M+Na)* = 335. -272- WO 99/54299 PCT/US99/07945 Example 82 (±)-(2R,3S,5R, 1 'R,2'S,3'S)-2-(1-Acetamido-2-hydroxy-3-methyl)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN.
O
t B u Boc O O 82A (±)-(2R,3S,5R,1'R,3'RS)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-oxo-3 methyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 42A, substituting (±)-(2R,3S,5R,1'R,2'R,3'RS)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'R) 1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl (yield: 12 mg, 63%).
CH
3
CH
3 AcHN., OtBu AcHN. c/OtBu OH OH - OH ' 0o H 0 82B (±)-(2R,3S.5R,1'R,2'S,3'S) and (±)-(2R,3S,5R,1'R,2'S,3'R)-1-t Butoxycarbonyl-2-(1-acetamido-2-hydroxy-3-methyl)pentyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compounds were prepared according to the method described in Example 42B, substituting (±)-(2R,3S,5R, 1'R,3'RS)-1l-t-butoxycarbonyl-2-(1 acetamido-2-oxo-3-methyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid -273- WO 99/54299 PCT/US99/07945 t-butyl ester (Rf= 0.5 and 0.65, 1:1, ethyl acetate: hexanes) in place of (2R,3S,5R, 1'R)-2-(1 -acetamido-2-oxo)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester to give (±)-(2R,3S,5R,1'R,2'S,3'S)-1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy-3-methyl)pentyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic acid t-butyl ester (Rf= 0.15, 1:1, ethyl acetate: hexanes) (yield: 6.0 mg, 50%) and (±)-(2R,3S,5R,1'R,2'S,3'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester(Rf= 0.10, 1:1, ethyl acetate: hexanes) (yield: 2.5 mg, 63%).
CH
3 AcHN. OH 'N l OH TFA 82C (±)-(2R,3S,5R,1'R,2'S,3'S)-2-(1-Acetamido-2-hydroxy-3-methyl)pentyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'S,3'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-methyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (Rf= 0.15, 1:1, ethyl acetate: hexanes) in place of (±) (2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 6.0 mg, 100%). 1 H NMR (DMSO-d 6 ) 6 7.78 (d, J=9.2Hz, 1H), 5.42 (m, 1H), 5.29 (t, J=10.3Hz, 1H), 4.08 (m, 1H), 3.96 (brt, 1H), 3.51 (m, 2H), 3.08 (m, 1H), 2.33 (m, 1H), 1.78 (s, 3H), 1.56 (d, J=6.3Hz, 3H), 1.52 (m, 1H), 1.40 (m, 1H), 1.29 (m, 1H), 1.21 (m, 1H), 0.84 (t, J=7.3Hz, 3H), 0.73 (d, J=6.9Hz, 3H) MS: (M-H)- = 311; (M+H) = 313, (M+Na) = 335. -274- WO 99/54299 PCT/US99/07945 Example 83 (±)-(2R,3S,5R, 1 'R,2'S,3'R)-2-(1 -Acetamido-2-hydroxy-3-methyl)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OH 'N HH OH TFA The title compound was prepared according to the method described in Example 41C, substituting(±)-(2R,3S,5R, 1 'R,2'S,3'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-methyl)pentyl-3-(cis-propen-1 -yl)-pyrrolid idine-5-carboxyl ic acid t-butyl ester (Rf= 0.10, 1:1 ethyl acetate: hexanes) in place of (±) (2R,3S,5R,1'R,2'S)- 1 -t-butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 2.5 mg, 100%). 1 H NMR (DMSO-d 6 ) 8 7.85 (d, J=8.7Hz, 1H), 5.45 (m, 1H), 5.29 (t, J=9.3Hz, 1H), 4.20 (m, 2H), 3.63 (t, J=8.3Hz, 1H), 3.42 (br d, 1H), 3.14 (m, 1H), 2.41 (m, 1H), 1.79 (s, 3H), 1.62 (m, 1H), 1.58 (d, J=5.4Hz, 3H), 1.43 (m, 2H), 1.0 (m, 1H), 0.88 (d, J=6.8Hz, 3H), 0.80 (t, J=7.3Hz, 3H) MS: (M-H) = 311; (M+H) = 313, (M+Na)* = 335. -275- WO 99/54299 PCT/US99/07945 Example 84 (±)-(2 R, 3S, 5 R, 1'R,2'S)-2-(1 -Acetamido-2-methoxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OtBu N H BocO
OCH
3 84A (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butviyl Ester (±)-(2R,3S,5R, 1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (22 mg, 0.05 mmole) was reacted with methyl iodide (0.016 mL, 0.25 mmole), potassium hydroxide (14 mg, 0.25 mmole) and 18-crown-6 (0.7 mg, 0.0025 mmole) in N,N-dimethylformamide (2 mL) at room temperature for 23 hours. Water (5 mL) was then added to the reaction mixture, followed by extraction with ether (2 x 10 mL). The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 66% ethyl acetate/hexanes to provide the title compound, as a colorless oil (yield: 5.2 mg, 23%). MS: (M+H) = 455, (M-H)- = 453. -276- WO 99/54299 PCT/US99/07945
CH
3 AcHN OH H H 0OH 3 O TFA 84B (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-methoxy)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 4.7 mg, 98%). 1 H NMR (DMSO-d6) 5 7.96 (d, J= 9.2Hz, 1H), 5.50 (m, 1H), 5.24 (m, 1H), 4.25 (m, 2H), 3.70 (m, 1H), 3.23 (s, 3H), 3.19 (m, 2H), 2.40 (m, 2H), 1.86 (s, 3H), 1.68 (m, 2H), 1.62 (dd, J= 7.0, 1.8Hz, 3H), 1.39 (m, 1H), 0.77 (t, J= 7.3Hz, 3H). MS: (M+H)Y= 299, (M+Na)+= 321, (M-H)- = 297 -277- WO 99/54299 PCT/US99/07945 Example 85 (±)-(2R, 3S,5R, 1'R,2'R)-2-(1-Acetamido-2-methoxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1=',
CH
3 AcHN tBu H, Boc" 5OCH 3 85A (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (17 mg, 0.04 mmole) was reacted with methyl iodide (28 mg, 0.19 mmole), potassium hydroxide (8 mg, 0.19 mmole) and 18-crown-6 (0.002 mmole) in N,N dimethylformamide (1.5 mL) at room temperature for 6 hours. Water (5 mL) was then added to the reaction mixture, followed by extraction with ether (2 x 10 mL). The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 50% ethyl acetate/hexanes to provide the title compound, (yield: 5 mg, 29%). MS: (M+H) =455, (M-H) =453 -278- WO 99/54299 PCT/US99/07945 AcHN. OH ~Nf H0
OCH
3 TFA TFA 85B (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-methoxy)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'R)-1 -t-butoxycarbonyl-2-(1 acetamido-2-methoxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 4 mg, 95%). 1H NMR (DMSO-d6) d 8.00(d, J=9.8HZ, 1H), 5.57(m, 1H), 5.35(m, 1H), 4.42(m, 1H), 4.28(m, 1H),3.95(m, 1H), 3.54(m, 1H), 3.28(s, 3H), 2.80(m, 1H), 2.30(m, 1H), 1.92(s, 3H), 1.65(m, 1H), 1.60(m, 3H), 1.43(m, 2H), 0.82(t, J=7.31HZ, 3H). MS: (M+H)+=299, (M-H)-=297 -279- WO 99/54299 PCT/US99/07945 Example 86 (±)-(2R, 3S,5R, 1'R,2'S)-2-(1-Acetamido-2-methoxy-3-methyl)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OH H Boc o
OCH
3 86A (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-methoxy-3 methyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound is prepared according to the method described in Example 84A, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-methyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester.
CH
3 AcHN. ,OH HH OCH 3 TFA 86B (±)-(2R,3S,5R,1'R,2'S)-2-( 1-Acetamido-2-methoxy-3-methyl)butyl-3-(cis propen-1 -ylv)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound is prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1 -t-butoxycarbonyl-2-(1 acetamido-2-methoxy-3-methyl)butyl-3-(cis-propen-l -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 -280- WO 99/54299 PCT/US99/07945 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. Example 87 (±)-(2R,3S,5R, 1 'R,2'R)-2-(1 -Acetamido-2-methoxy-3-methyl)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OH N -d H Boc O
OCH
3 86A (±)-(2R.,3S.,5R, 1'R,2'R)-1l-t-Butoxycarbonyl-2-(1l-acetamido-2-methoxy-3 methyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 84A, substituting (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(l acetamido-2-hydroxy-3-methyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 6.8 mg, 33%). MS: (M+H) = 469, (M+Na) += 491, (M-H)- = 467. -281- WO 99/54299 PCT/US99/07945
CH
3 AcHN. ,-OH N H 0
OCH
3 TFA 87B (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-methoxy-3-methyl)butyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxy-3-methyl)butyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester, ester (yield: 6.6 mg, 100%). 1 H NMR (DMSO-de) 8 7.65 (d, J= 9.2Hz, 1H), 5.43 (m, 1H), 5.23 (m, 1H), 4.42 (m, 1H), 4.37 (m, 1H), 3.56 (m, 1H), 3.46 (s, 3H), 3.17 (m, 2H), 2.44 (m, 1H), 1.80 s, 3H), 1.78 (m, 1H), 1.70 (m, 1H), 1.57 (dd, J= 6.7, 1.2Hz, 3H), 0.94 (d, J= 6.7Hz, 3H), 0.82 (d, J= 6.7Hz, 3H). MS: (M+H) = 313, (M+Na)+ = 335, (M-H)- = 311. -282- WO 99/54299 PCT/US99/07945 Example 88 (±)-(2R,3S,5R. 1'R,2'S)-2-(1 -Acetamido-2-methoxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OH 'N? B 0
OCH
3 88A (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxy)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 84A, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 11.9 mg, 36%). MS: (M+H)*= 469, (M+Na)+= 491, (M-H)- = 467.
CH
3 AcHN. , OH 'N H 0
OCH
3 TFA 88B (±)-(2R.3S,5R,1'R,2'S)-2-(1-Acetamido-2-methoxy)pentyl-3-(cis-propen-1 vl)-pvrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1 'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-methoxy)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R, 1 'R,2'S)-1-t-butoxycarbonyl-2-(1-acetamido -283- WO 99/54299 PCT/US99/07945 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. (yield: 11.5 mg, 100%). 1 H NMR (DMSO-d 6 ) 6 7.95 (d, J= 9.8Hz, 1H), 5.49 (m, 1H), 5.23 (m, 1H), 4.25 (m, 2H), 3.68 (m, 1H), 3.24 (s, 3H), 3.22 (m, 1H), 3.18 (m, 1H), 2.40 (m, 1H), 1.85 (s, 3H), 1.66 (m, 1H), 1.62 (m, 3H), 1.58 (m, 1H), 1.38 (m, 1H), 1.27 (m, 2H), 0.86 (t, J= 7.3Hz, 3H). MS: (M+H) = 313, (M+Na)+=335, (M-H)- = 311. Example 89 (±)-(2R,3S,5R, 1'R.2'R)-2-(1-Acetamido-2-methoxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHNT OH Boc 0
OCH
3 89A (±)-(2R,3S,5R, 1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxy)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 84A, substituting (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R, 3S, 5R, 1'R,2'S)-1 -t-butoxycarbonyl-2-(1 -acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 4.3 mg, 21%). MS: (M+H)*= 469, (M+Na) = 491, (M-H)-= 467. -284- WO 99/54299 PCT/US99/07945 AcHOH
CH
3
OCH
3 AcHN- ,OH H 0
OCH
3 TFA 89B (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-methoxy)pentyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-methoxy)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 4.8 mg, 100%). 1 H NMR (DMSO-d 6 ) 6 7.70 (d, J= 9.8Hz, 1H), 5.45 (m, 1H), 5.24 (m, 1H), 4.40 (m, 1H), 4.25 (m, 1H), 3.57 (t, J= 8.5Hz, 1H), 3.40 (m, 1H), 3.35 (s, 3H), 3.17 (m, 1H), 2.42 (m, 1H), 1.82 (s, 3H), 1.69 (m, 1H), 1.56 (dd, J= 7.1, 1.2Hz, 3H), 1.24 (m, 4H), 0.88 (t, J= 7.0Hz, 3H). MS: (M+H) = 313, (M+Na)+= 335, (M-H)- = 311 -285- WO 99/54299 PCT/US99/07945 Example 90 (±)-(2R,3S,5R. 1'R,2'S)-2-(1-Acetamido-2-methoxy-2-allyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN- OtBu 'N f H Boc o
OCH
3 90A (±)-(2R,3S,5R, 1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-methoxy-2 allyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butvl Ester The title compound was prepared according to the method described in Example 84A, substituting (±i)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-allyl)ethyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 8 mg, 31%). MS: (M+H)+=467, (M-H)-=465 90B (±)-(2R.3S,5R,1'R,2'S)-2-(1-Acetamido-2-methoxy-2-allyl)ethyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxy-2-allyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, I'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester, ester (yield: 6 mg, 96%). 1 H NMR (DMSO-d 6 ) 6 8.02(d, J=8.6HZ, 1H),5.75 (m, 1H), 5.51(m, 1H), 5.24(m, 1H), 5.05(m, 2H), 4.27(m, 1H), 4.22(m, 1H), 3.74(m, 2H), 3.26(s, 3H), -286- WO 99/54299 PCT/US99/07945 3.18(m, 1H), 2.47(m, 1H), 2.39(m, 1H), 2.17(m, 1H), 1.87(s, 3H), 1.67(m, 1H),1.63(dd, J=6.71, 1.23 HZ, 3H). MS: (M+H) =311, (M-H)- =309 Example 91 (±)-(2R,3S,5R,1'R.2'R)-2-(1-Acetamido-2-methoxy-2-allyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt CH 3 AcHN.. .OtBu 'N v OCH 3 91A (±)-(2R,3S,5R,1'R.2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-methoxy-2 allyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 84A, substituting (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-allyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 4.0 mg, 16%). MS: (M+H) =467, (M-H) =465 -287- WO 99/54299 PCT/US99/07945 OH AcHN. OH N H H 0
"OCH
3 91B (±)-(2R.3S,5R,1'R,2'R)-2-(1-Acetamido-2-methoxy-2-allyl)ethyl-3-(cis propen-1 -ylv)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxy-2-allyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 3 mg, 96%). 'H NMR (DMSO-d 6 ) 8 7.75 (d, J=9.2 HZ, 1H), 5.75(m, 1H), 5.47(m, 1H), 5.24(m, 1 H), 5.06(m, 2H), 4.42(m, 1H), 4.25(m, 1H), 3.58(m, 1H), 3.50(m, 1H), 3.37(s, 3H), 3.17(m, 1H), 2.42(m, 1H), 2.36(m, 1H), 1.83(s, 3H), 1.71(m, 1H), 1.55(dd, J=6.73, 1.83 HZ, 3H) MS: (M+H)=311, (M-H) =309 -288- WO 99/54299 PCT/US99/07945 Example 92 (±)-(2R, 3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 CH 3 AcHN. OtBu AcHN, OtBu HN ' HN '
SB
oc O Bo c O OH OH 92A (±)-(2R,3S,5R,1'R,2'S) and (±)-(2R,3S,5R,1 'R,2'R)-1l-t-Butoxycarbonyl-2 (1-acetamido-2-hydroLxy-4-vinyl)butyvl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compounds were prepared according to the method described in Example 41B, substituting 1-buten-4-yl magnesium bromide for ethyl magnesium bromide to provide (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy-4-vinyl)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0030 g, 6%) and (±)-(2R,3S,5R,1'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0145 g, 28%). (±)-(2R,3S,5R,1'R,2'S) MS: (M+H)*=467, (M+Na)+=489, (2M+Na) =955, (M-H)-=465. (±)-(2R,3S,5R, 1'R,2'R)- MS: (M+H) =467, (M+Na)*=489, (2M+Na) =955, (M-H)-=465. -289- WO 99/54299 PCT/US99/07945
OH
3 AcHN. OH N H 0 OH TFA 92B (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-4-vinyl)butyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0027 g, 100%). 1 H NMR (DMSO-d 6 ) 5 8.93 (bs, 1H), 7.90 (d, J=9.2 Hz, 1H), 5.80 (m, 1H), 5.48 (m, 1H), 5.28 (m, 1H), 5.00 (dd, J=17.1, 1.8Hz, 1H), 4.94 (dd, J=10.4,1.8Hz, 1H), 4.29 (bt, J=8.3Hz, 1H), 4.03 (m, 1H), 3.71 (m, 1H), 3.49 (m, 1H), 3.15 (quint., J=8.5Hz, 1H), 2.41 (dt, J=12.8,7.3Hz, 1H), 2.16 (M, 1H), 2.05 (m, 1H), 1.83 (s, 3H), 1.79-1.75 (m, 1H), 1.64 (m, 1H), 1.58 (dd, J=6.7,1.8Hz, 3H), 1.34 (m, 2H). MS: (M+H) = 311, (M+Na)* = 333, (M-H) = 309, (M+CF 3 COO-)=423 -290- WO 99/54299 PCT/US99/07945 Example 93 (±)-(2R,3S,5R, 1 'R.2'R)-2-(1 -Acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen-1 yl)-pyvrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN, OH N H * 0 OH TFA 93A (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-4-vinyl)butyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0027 g, 100%). 1 H NMR (DMSO-d 6 ) 8 7.68 (d, J=9.6 Hz, 1H), 5.81 (m, 1H), 5.48 (m, 1H), 5.25 (m, 1H), 5.01 (dd, J=17.1, 1.8Hz, 1H), 4.95 (dd, J=10.3,1.7Hz, 1H), 4.43 (t, J=8.5Hz, 1H), 4.10 (m, 1H), 3.74 (m, 1H), 3.56 (t, J=8.9Hz, 1H), 3.16 (quint., J=8.9Hz, 1H), 2.42 (dt, J=12.8,7.3Hz, 1H), 2.11 (M, 1H), 2.07 (m, 1H), 1.83 (s, 3H), 1.72 (dt, J=12.8, 9.8Hz, 1H), 1.55 (dd, J=6.7,1.8Hz, 3H), 1.5-1.35 (m, 2H). MS: (M+H) = 311, (M+Na) = 333, (M-H)- = 309, (M+CF 3 COO-)-=423, (2M-H)=619. -291- WO 99/54299 PCT/US99/07945 Example 94 (±)-(2R,3S,5R,1'R,2'S,3'S)-2-(1 -Acetamido-2-methoxy-3-methyl)pentyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OH N H Boc
OCH
3 94A (±)-(2R,3S,5R,1'R,2'S,3'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-methoxy 3-methyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound is prepared according to the method described in Example 84A, substituting (±)-(2R,3S,5R,1'R,2'S,3'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-methyi)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester.
CH
3 AcHN. OH 'N H 0
OCH
3 TFA 94B (±)-(2R,3S.5R,1'R.2'S,3'S)-2-(1-Acetamido-2-methoxy-3-methyl)pentyl-3 (cis-propen-1 -ylv)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound is prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'S,3'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxy-3-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 -292- WO 99/54299 PCT/US99/07945 acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester. Example 95 (±)-(2R,3S,5R,1'RS)-2-(1-Acetamido-2-oxo-2-heptafluoropropyl)ethyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN OtBu F F N H Boc O
F
3 C O FF 95A (±)-(2R,3S,5R,1 'RS)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-oxo-3 heptafluoropropyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 42A, substituting (±)-(2R,3S,5R, 1'R,2'RS)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-heptafluoropropyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester for (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolid i ne-5-carboxylic acid t-butyl ester (yield: 6.8 mg, 88%). MS: (M+H)*=579, (M-H)=577. -293- WO 99/54299 PCT/US99/07945
CH
3 AcHN OH F F 'N HH
F
3 C O F F TFA 95B (±)-(2R. 3S,5R, 1'RS)-2-(1 -Acetamido-2-oxo-2-heptafluoropropyl)ethyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'RS)-1 -t-butoxycarbonyl-2-(1 acetamido-2-oxo-3-heptafluoropropyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 0.0037 g, 100%). MS: (M+H)*=423, (M-H)-=421. Example 96 -294- WO 99/54299 PCT/US99/07945 (±)-(2R,3S, 5R, 1'RS)-2-(1 -Acetamido-2-oxo-2-heptafluoropropyl)ethyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN Ot{tBu F F N ' FF H BocO
F
3 0 0 O FF 96A (±)-(2R,3S,5R,1 'RS)-1-t-Butoxycarbonyl-2-(1-Acetamido-2-oxo-3 heptafluoropropyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 42A, substituting (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-heptafluoropropyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester for (±)-(2R,3S,5R, 1'R,2'R)-1 -t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 6.8 mg, 88%). MS: (M+H)*=579, (M-H)=577.
CH
3 AcHN N OH FF N H H 0
F
3 C O F F TFA 96B (±)-(2R.3S,5R, 1'RS)-2-(1-Acetamido-2-oxo-2-heptafluoropropyl)ethyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'S)-1 -t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-3-heptafluoropropyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1-t-butoxycarbonyl -295- WO 99/54299 PCT/US99/07945 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 0.0037 g, 100%). MS: (M+H)*=423, (M-H)-=421. Example 97 (±)-(2R,3S,5R, 1'R)-2-(1-Acetamido-2-oxo)pentyl-3-(cis-propen-1-yl)-pyrrolidine 5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN OtBu ~HBoco 0O 0 97A (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-oxo)pentvl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 42A, substituting (±)-(2R,3S,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hdroxy)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 14 mg, 58%). MS: (M+H) = 453, (M+Na)* = 475; (M-H)- = 451. -296- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH N 0 0 TFA 97B (±)-(2R,3S,5R, 1 'R)-2-(1-Acetamido-2-oxo)pentyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-oxo)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 1.4 mg, 28%). 1 H NMR (DMSO-d 6 ) 6 8.31 (d, J=8.3Hz, 1H), 5.40 (m, 1H), 5.19 (brt, 1H), 4.26 (t, J=6.8Hz, 1 H), 3.63 (t, J=8.3Hz, 1H), 3.35 (m, 1H), 2.97 (m, 1H), 2.45 (m, 1H), 2.34 (dt, J=3.4, 7.4Hz, 1H), 2.20 (m, 1H), 1.84 (s, 3H), 1.58 (dd, J=2, 4.3Hz, 3H), 1.43 (m, 3H), 0.82 (t, J=7.3Hz, 3H) MS: (M-H) = 295; (M+H) = 297, (M+Na) + = 319. -297- WO 99/54299 PCT/US99/07945 Example 98 (±)-(2R. 3S, 5R, 1'R)-2-(1 -Acetamido-2-oxo)butvl-3-(cis-propen-1 -yl)-pyrrolid i ne-5 carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcH N - . OH H H 0 TFA The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R)-1-t-butoxycarbonyl-2-(1 acetamido-2-oxo)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester prepared in Example 42A in place of (±)-(2R,3S,5R,1'R,2'S)-1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 5.0 mg, 100%). 1H NMR (DMSO-d 6 ) 5 8.52 (d, J= 8.6Hz, 1H), 5.47 (m, 1H), 5.15 (m, 1H), 4.54 (m, 1H), 4.39 (dd, J= 11.0, 6.7Hz, 1H), 3.84 (t, J= 9.2Hz, 1H), 3.17 (m, 1H), 2.50 (m, 1H), 2.38 (m, 1H), 2.33 (m, 1H), 1.83 (s, 3H), 1.63 (m, 1H), 1.58 (dd, J= 6.7, 1.8Hz, 3H), 0.94 (t, J= 7.5Hz, 3H). MS: (M+H) = 283, (M+Na) = 305, (M-H)- = 281. Examples 99-115 The title compounds were prepared according to the methods described in Examples 20 and 40-42 by substituting the respective reactants. -298- WO 99/54299 PCT/US99/07945 Example 99
CH
3 AcHN. .. OH 'N HH 0 0 TFA (±)-(2R,3S,5R,1'R)-2-(1 -Acetamido-2-oxo-2-allyl)ethyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 8 8.38 (d, J= 8.5Hz, 1H), 5.73 (m, 1H), 5.37 (m, 1H), 5.05 (m, 3H), 4.32 (t, J= 7.9Hz, 1H), 3.90 (m, 1H), 3.49 (m, 1H), 3.13 (m, 2H), 2.98 (m, 1H), 3.18 (m, 1H), 1.78 (s, 3H), 1.51 (dd, J= 5.5, 1.2Hz, 3H), 1.44 (m, 1H). MS: (M+H) 4 = 295, (M-H)- = 293. Example 100 o AcHN. OH HH 0 0 TFA (±)-(2R,3S,5R,1 'R)-2-(1-Acetamido-2-oxo-3-methyl)butyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 8 8.64 (d, J= 8.5Hz, 1H), 5.59 (m, 1H), 5.08 (d, J= 17.1Hz, 1H), 5.02 (d, J= 9.8Hz, 1H), 4.65 (t, J= 8.6Hz, 1H), 4.32 (m, 1H), 3.82 (t, J= 9.2Hz, 1H), 2.82 (m 2H), 2.36 (m, 1H), 1.83 (s, 3H), 1.80 (m, 1H), 1.03 (d, J= 6.7Hz, 3H), 0.97 (d, J= 6.7Hz, 3H). MS: (M+H)*= 283, (M+Na)* = 305, (M-H)- = 281. -299- WO 99/54299 PCT/US99/07945 Example 101 = AcHN. OH 0 0 0 TFA (±)-(2R,3S. 5R, 1 'R)-2-(1 -Acetamido-2-oxo)propyl-3-vinyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 6 8.96 (d, J= 7.9Hz, 1H), 5.71 (m, 1H), 5.27 (d, J=17.7Hz, 1H), 5.17 (d, J= 11.0Hz, 1H), 4.38 (m, 1H), 4.29 (m, 1H), 3.81 (m, 1H), 2.61 (m, 1H), 2.22 (m, 1H), 2.13 (s, 3H), 2.01 (s, 3H), 1.24 (m, 1H). MS: (M+H) = 255, (M+Na) + = 277, (M-H)- = 253. Example 102 = AcHN OH H 0 0 TFA (±)-(2R,3S.5R, 1'R)-2-(1 -Acetamido-2-oxo)butyl-3-vinyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 8 8.61 (d, J= 8.5Hz, 1H), 5.60 (m, 1H), 5.10 (d, J= 17.7Hz, 1H), 5.03 (dd, J= 10.4, 1.2Hz, 1H), 4.54 (t, J= 8.5Hz, 1H), 4.38 (dd, J= 11.0, 6.7Hz, 1H), 3.86 (m, 1H), 2.84 (m, 1H), 2.52 (m, 1H), 2.37 (m, 2H), 1.85 (s, 3H), 1.82 (m, 1H), 0.94 (t, J= 7.0Hz, 3H). MS: (M+H) = 269, (M+Na)* = 291, (M-H)- = 267. -300- WO 99/54299 PCT/US99/07945 Example 103 AcHN- <OH H O 0 TFA (±)-(2R. 3S, 5R, 1'R)-2-(1 -Acetamido-2-oxo)pentvlI-3-viny-pyrrolidine-5-carboxlic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 8 8.60 (d, J= 9.7Hz, 1H), 5.60 (m, 1H), 5.07 (m, 2H), 4.65 (m, 1H), 4.54 (m, 1H), 4.38 (m, 1 H), 3.86 (m, 1H), 2.84 (m, 1H), 2.45 (m, 1H), 2.36 (m, 1H), 1.86 (s, 3H), 1.82 (m, 1H), 1.47 (m, 2H), 0.87 (t, J= 5.8Hz, 3H). MS: (M+H)*= 283, (M+Na) += 305, (M-H)- = 281. Example 104 AcHN. OH N OH 0 OH TFA (±)-(2R,3S.5R,1'R)-2-(1 -Acetamido-2-hydroxy)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 6 8.00 (d, J= 9.9Hz, 1H), 5.63 (m, 1H), 5.08 (m, 1H), 4.98 (m, 1H), 4.35 (m, 1H), 4.25 (m, 1H), 4.08 (m, 1H), 3.55 (m, 1H), 3.45 (m, 1H), 3.38 (m, 1H), 2.83 (m, 1H), 2.33 (m, 1H), 1.78 (s, 3H). MS: (M+H)* = 243, (M+Na)+= 265, (M-H)- = 241. -301- WO 99/54299 PCT/US99/07945 Example 105 = AcHN OH OH TFA (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)propyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 8 7.96 (d, J= 9.7Hz, 1H), 5.74 (m, 1H), 5.12 (m, 1H), 5.03 (m, 1H), 4.27 (m, 1H), 3.96 (m, 1H), 3.77 (m 1H), 3.65 (m, 1H), 2.87 (m, 1H), 2.38 (m, 1H), 1.82 (s, 3H), 1.80 (m, 1H), 1.08 (d, J= 6.0Hz, 3H). MS: (M+H)
+
= 257, (M+Na)
+
= 279, (M-H) = 255. Example 106 = AcHN OH OH 0 TFA (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamido-2-hydroxy)butyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 87.99 (d, J= 9.0Hz, 1H), 5.75 (m, 1H), 5.13 (d, J= 17.1Hz, 1H), 5.04 (d, J= 10.5Hz, 1H), 4.27 (t, J= 8.4Hz, 1H), 4.04 (m, 1H), 3.78 (m, 1H), 3.48 (m, 1H), 2.89 (m, 1H), 2.40 (m, 1H), 1.88 (m, 1H), 1.85 (s, 3H), 1.54 (m, 1H), 1.28 (m, 1H), 0.86 (t, J= 7.2Hz, 3H). MS: (M+H) = 271, (M+Na)+ = 293, (M-H)- = 269. -302- WO 99/54299 PCT/US99/07945 Example 107 OH TFA (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)pentyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 8 7.99 (d, J= 9.9Hz, 1H), 5.75 (m, 1H), 5.08 (m, 2H), 4.28 (m, 1 H), 4.03 (m, 1H), 3.77 (m, 1H), 3.52 (m, 1H), 2.88 (m, 1H), 2.40 (m, 1H), 1.86 (s, 3H), 1.75 (m, 1H), 1.45 (m, 2H), 1.25 (m, 2H), 0.87 (t, J= 5.9Hz, 3H). MS: (M+H) = 285, (M+Na) = 307, (M-H) = 283. Example 108 = AcHN. OH OH 0 TFA (±)-(2R,3S,5R, 1'R.2'S)-2-(1-Acetamido-2-hydroxy-3-methyl)butyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 5 7.97 (d, J= 9.3Hz, 1H), 5.75 (m, 1H), 5.12 (d, J= 17.1Hz, 1H), 5.04 (d, J= 11.2Hz, 1H), 4.24 (m, 1H), 4.13 (m, 1H), 3.74 (dd, J= 9.8, 6.1Hz, 1H), 3.44 (dd, J= 10.3, 2.0Hz, 1H), 2.87 (m, 1H), 2.40(m, 1H), 1.84 (m, 1H), 1.83 (s, 3H), 1.75 (m, 1H), 0.89 (d, J= 6.8, 3H), 0.75 (d, J= 6.8Hz, 3H). MS: (M+H) = 285, (M+Na)* = 307, (M-H) = 283. -303- WO 99/54299 PCT/US99/07945 Example 109 AcHN. OH N OH O TFA (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamido-2-hydroxy-2-cyclopropyl)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 8 7.81 (d, J= 10.0Hz, 1H), 5.73 (m, 1H), 5.05 (m, 2H), 4.39 (m, 1H), 4.20 (m 1H), 3.90 (m, 1H), 3.61 (m, 1H), 3.08 (m, 1H), 2.86 (m, 1H), 2.42 (m, 1H), 1.85 (s, 3H), 0.88 (m, 1H), 0.45 (m, 1H), 0.35 (m, 2H), 0.11 (m, 1H). MS: (M+H) = 283, (M+Na)*= 305, (M-H)- = 281. Example 110 = AcHN OH ~OH TFA (±)-(2R,3S,5R, 1 'R,2'R)-2-(1 -Acetamido-2-hydroxy)propyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 6 7.77 (d, J= 9.7Hz, 1H), 5.72 (m, 1H), 5.07 (m, 2H), 4.40 (m, 1H), 4.03 (m, 1H), 3.95 (m 1H), 3.57 (m, 1H), 2.86 (m, 1H), 2.43 (m, 1H), 1.88 (m, 1H), 1.84 (s, 3H), 1.04 (d, J= 6.0Hz, 3H). MS: (M+H) = 257, (M+Na) = 279, (M-H) = 255. -304- WO 99/54299 PCT/US99/07945 Example 111 Ac OH "OH 0 TFA (±)-(2R,3S,5R, 1'R,2'R)-2-(1 -Acetamido-2-hydroxy)butyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 87.72 (d, J= 9.8Hz, 1H), 5.73 (m, 1H), 5.08 (d, J= 17.1Hz, 1H), 5.03 (d, J= 10.4Hz, 1H), 4.41 (m, 1H), 4.13 (m, 1H), 3.68 (m, 1H), 3.63 (m, 1H), 2.88 (m, 1H), 2.44 (m, 1H), 1.90 (m, 1H), 1.83 (s, 3H), 1.38 (m, 2H), 0.84 (t, J= 7.3Hz, 3H). MS: (M+H) = 271, (M+Na) = 293, (M-H)- = 269. Example 112 AcHN'. OH1 "OH O '00 TFA (±)-(2R,3S,5R. 1'R,2'R)-2-(1 -Acetamido-2-hydroxy)pentyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 8 7.72 (d, J= 9.9Hz, 1H), 5.72 (m, 1H), 5.06 (m, 2H), 4.42 (m, 1H), 4.09 (m, 1H), 3.77 (m, 1H), 3.61 (m, 1H), 2.87 (m, 1H), 2.43 (m, 1H), 1.90 (m, 1H), 1.83 (s, 3H), 1.37 (m, 2H), 1.27 (m, 2H), 0.87 (t, J= 5.9Hz, 3H). MS: (M+H) = 285, (M+Na)*= 307, (M-H) = 283. -305- WO 99/54299 PCT/US99/07945 Example 113 AcHN. OH N HH 'OH O TFA (±)-(2R,3S,5R.1'R,2'R)-2-(1 -Acetamido-2-hydroxy-3-methyl)butyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d6) 68 7.71 (d, J= 9.3Hz, 1H), 5.70 (m, 1H), 5.08 (d, J= 17.1Hz, 1H), 5.03 (d, J= 10.3Hz, 1H), 4.42 (m, 1H), 4.25 (m, 1H), 3.61 (m, 1H), 3.35 (dd, J= 8.3, 2.5Hz, 1H), 2.90 (m, 1H), 2.44 (m, 1H), 1.92 (m, 1H), 1.82 (s, 3H), 1.58 (m, 1H), 0.95 (d, J= 6.8Hz, 3H), 0.79 (d, J= 6.4Hz, 3H). MS: (M+H) += 285, (M+Na) + = 307, (M-H)- = 283. Example 114 AcHN. H OH 'OH O TFA (±)-(2R,3S,5R. 1 'R,2'R)-2-(1-Acetamido-2-hydroxy-2-cyclopropyvl)ethyl-3-vinyvl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 8 7.94 (d, J= 9.6Hz, 1H), 5.76 (m, 1H), 5.12 (m, 2H), 4.40 (m, 1H), 4.21 (m, 1H), 3.90 (m, 1H), 3.53 (m, 1H), 3.13 (m, 1H), 2.81 (m, -306- WO 99/54299 PCT/US99/07945 1H), 2.25 (m, 1H), 1.87 (s, 3H), 0.90 (m, 1 H), 0.47 (m, 1 H), 0.37 (m, 2H), 0.15 (m, 1 H). MS: (M+H) = 283, (M+Na) = 305, (M-H)- = 281. Example 115 AcHN. OH HH 'OH O S TFA (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy-4-methyl)pentyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 6 7.71 (d, J= 9.7Hz, 1H), 5.83 (m, 1H), 5.06 (d, J= 17.1Hz, 1H), 5.02 (d, J= 10.3Hz, 1H), 4.41 (m, 1H), 4.06 (m, 1H), 3.83 (m, 1H), 3.59 (t, J= 8.8Hz, 1H), 2.84 (m, 1H), 2.42 (m, 1H), 1.90 (m, 1H), 1.82 (s, 3H), 1.71 (m, 1H), 1.34 (m, 1H), 1.07 (m, 1H), 0.89 (d, J= 6.8Hz, 3H), 0.86 (d, J= 6.3Hz, 3H). MS: (M+H) = 299, (M+Na) = 321, (M-H)- = 297. -307- WO 99/54299 PCT/US99/07945 Example 116 (±)-(2R,3S.5 R,1'R)-2-(1-Acetamido-2-hydroxy-2-methvl)propyl-3-vinvl-pyrrolidine 5-carboxylic Acid Trifluoroacetic Acid Salt AcHN OtBu 'N i H Boc" OH 0 116A (±)-(2R,3S,5R, 1 'R)-t-Butoxycarbonyl-2-(1-Acetamido-2-hydroxy-2 methyl)propyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R,1'R)-t-Butoxycarbonyl 2-(1-acetamido-2-oxo)propyl-3-vinyl pyrrolidine-5-carboxylic acid t-butyl ester (11 mg, 0.027 mmol) was reacted with methyl magnesium bromide (3 M) ( 0.05mL, 0.134 mmol) in THF (2 mL) at 25 oC for 2 hours. The reaction was quenched with saturated aqueous ammonium chloride (2 mL) and water (2 mL) followed by extraction using dichloromethane (2 X 5 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 2/1: ethyl acetate/hexane to provide the title compound (yield: 1.9 mg, 17%). MS: (M+H)*=427, (M-H)- =425 -308- WO 99/54299 PCT/US99/07945 AcHN OH N OH O TFA 116B (±)-(2R, 3S,5R, 1'R)-2-(1 -Acetamido-2-hydroxy-2-methyl)propyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-methyl)propyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 1.6 mg, 99%). 1 H NMR (DMSO-d 6 ) 6 7.70(d, J=9.9Hz, 1H), 5.75(m, 1H), 5.02(m, 2H), 4.37(m, 1H), 4.15(m, 1H), 3.61(m, 1H), 2.78(m, 1H), 2.41(m, 1H), 1.81(s, 3H), 1.20(s, 3H), 1.12(s, 3H) MS: (M+H) =271, (M+23) =293, (M-H)- =269 -309- WO 99/54299 PCT/US99/07945 Example 117 (±)-(2R,3S,5R, 1'R)-2-(1 -Acetamido-2-hydroxy-2-ethyl)butyl-3-vinyl-pyrrolid ine-5 carboxylic Acid Trifluoroacetic Acid Salt = AcHN. "1OtBu HN H Boc O OH 117A (±)-(2R,3S,5R, 1 'R)-t-Butoxycarbonyl-2-(1-Acetamido-2-hydroxy-2 ethyl)butyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R,1'R)-t-Butoxycarbonyl-2-(1 -acetamido-2-oxo)butyl-3-vinyl pyrrolidine-5-carboxylic acid t-butyl ester (37mg, 0.087 mmol) was reacted with ethyl magnesium bromide (3 M) ( 0.15mL, 0.44mmol) in THF (5 mL) at 25 oC for 2 hours. The reaction was quenched with saturated aqueous ammonium chloride 5 mL) and water (5 mL) followed by extraction using dichloromethane (2 X 10 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 2/1: ethyl acetate/hexane to provide the title compound (yield: 14 mg, 35%). MS: (M+H) = 455, (M-H)- =453 -310- WO 99/54299 PCT/US99/07945 = AcHN OH 'N HH OH O TFA 116B (±)-(2R,3S,5R, 1'R)-2-(1-Acetamido-2-hydroxy-2-ethyl)butyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-ethyl)butyl-3vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1 'R,2'S)-1-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 5.8 mg, 98%). 1 H NMR (DMSO-d 6 ) 8 7.62(d, J=9.6HZ, 1H), 5.75(m, 1H), 5.03(m, 2H), 4.39(m, 1H), 4.31(m, 2H), 3.87(m, 1H), 3.38(m, 1H), 2.88( m, 1H), 2.40(m, 1H), 1.83(s, 3H), 1.55-1.30(m, 4H), 0.86(m, 6H) MS: (M+H)* =299, (M-H) =297 -311- WO 99/54299 PCT/US99/07945 Example 118 (±)-(2R.3S,5R, 1 'S)-2-(1 -Acetamido)allyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxvlic Acid Trifluoroacetic Acid Salt
CH
3 OtBu AcHN. O t 118A (±)-(2R,3S,5R, 1 'S)-1-t-Butoxycarbonyl-2-(1 -acetamido)alyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 20K, substituting (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido 2-formyl)methyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 formyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 15.3 mg, 61.4%). MS: (M+H) = 409.
CH
3 AcHN. OH NH H TFA 118B (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido)allyl-3-(cis-propen-1 -ylv)-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'S)-1 -t-butoxycarbonyl-2-(1 acetamido)allyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1-acetamido-2 -312- WO 99/54299 PCT/US99/07945 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 13.1 mg, 100%). 1 H NMR (DMSO-d6): 81.58 (dd, 3H), 1.74 (dt, 1H), 1.88 (s, 3H), 2.41 (dt, 1H), 3.17 (m, 1H), 3.56 (dd, 1H), 4.35 (dd, 1H), 4.70 (dd, 1H), 5.22-5.30 (m, 3H), 5.51 (m, 1H), 5.82 (m, 1H), 8.15 (d, 1H), 9.18 (brs, 2H). MS: (M+H) = 253. Example 119 (±)-(2R,3S,5R, 1 'S)-2-(1-Acetamido-2-(cis and trans)buten-1 -yl)-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt /=" CH3 Ac OtBU 119A (±)-(2R,3S,5R. 1 'S)-l-t-Butoxycarbonyl-2-(1 -Acetamido-2-(cis and trans)buten-1 -yl)-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 20K, substituting (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido 2-formyl)methyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, I'S)-l-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 formyl-pyrrolidine-5-carboxylic acid t-butyl ester and ethyltriphenylphosphonium bromide for methyltriphenylphosphonium bromide (yield: 12.4 mg, 48.2%). MS: (M+H) = 423 -313- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH Nf
H
3 C- H 0 TFA 119B (±)-(2R.3S, 5R, 1 'S)-2-(1 -Acetamido-2-(cis and trans) b uten-1 -yl)-3-(cis propen-1 -ylv)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'S)-1 -t-butoxycarbonyl-2-(1-acetamido 2-(cis and trans)buten-1-yl)-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 11.8 mg, 100%). 'H NMR (DMSO-d 6 ): 61.63 (dd, 3H), 1.66 (dd, 3H), 1.74 (m, 1H), 1.88 (s, 3H), 2.41 (dt, 1H), 3.17 (m, 1H), 3.50 (dd, 1H), 4.34 (dd, 1H), 4.95 (m, 1H), 5.23 (m, 1H), 5.39 (m, 1H), 5.53 (m, 1H), 5.68 (m, 1H), 8.21 (d, 1H), 9.18 (br s, 2H). MS: (M+H)
+
= 267 -314- WO 99/54299 PCT/US99/07945 Example 120 (±)-(2R,3S,5R,1'S)-2-(1 -Acetamido-3,3-dimethyl)allyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OtBu NO B 0 120A (+)-(2R,3S,5R, 1'S)-l-t-Butoxycarbonyl-2-(1-acetamido-3,3-dimethyl)allyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 20K, substituting (±)-(2R,3S,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-formyl)methyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1 'S)-l-t-butoxycarbonyl-2-(1 -acetamido-3-methyl)butyl-3 formyl-pyrrolidine-5-carboxylic acid t-butyl ester and isopropyltriphenylphosphonium bromide for methyltriphenylphosphonium bromide (yield: 8.2 mg, 25.9%). MS: (M+H)*= 437
CH
3 AcHN. OH 'N / HH O TFA 120B (±)-(2R,3S,5R, 1 'S)-2-(1-Acetamido-3,3-dimethvl)allyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido 3,3-dimethyl)aIlyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in -315- WO 99/54299 PCT/US99/07945 place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester, ester (yield: 7.5 mg, 100%). 1 H NMR (DMSO-d6): 61.53 (dd, 3H), 1.57 (s, 3H), 1.61 (s, 3H), 1.66 (m, 1H), 1.77 (s, 3H), 2.32 (dt, 1H), 3.07 (m, 1H), 3.39 (dd, 1H), 4.26 (m, 1H), 4.75 (m, 1H), 5.07 (d, 1H), 5.15 (m, 1H), 5.44 (m, 1H), 8.06 (d, 1H). MS: (M+H)* = 281. Example 121 (±)-(2R,3S,5R, 1 'S)-2-(1-Acetamido-2-(cis and trans)penten-1 -vyl)-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 OtBu AcH N. Hj r Boc 6 121A (±)-(2R,3S,5R, 1'S)-1l-t-Butoxycarbonyl-2-(1-Acetamido -2-(cis and trans)penten-1 -yl)-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example substituting (±)-(2R,3S,5R,1'S)-1 -t-butoxycarbonyl-2-(1-acetamido-2 formyl)methyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 formyl-pyrrolidine-5-carboxylic acid t-butyl ester and n-butyl triphenylphosphonium bromide for methyltriphenylphosphonium bromide (yield: 21.0 mg, 66.2%). MS: (M+H)* = 437. -316- WO 99/54299 PCT/US99/07945
CH
3 AcHN- OH Nf H. H o TFA 121B (±)-(2R, 3S,5R, 1'S)-2-(1-Acetamido-2-(cis and trans)penten-1 -yl)-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'S)-1 -t-butoxycarbonyl-2-(1 -acetamido 2-(cis and trans)penten-1-yl)-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. ester (yield: 16.0 mg, 98.1%). 1 H NMR (DMSO-d6): 8 0.93 (t, 3H), 1.62 (dd, 3H), 1.75 (m, 1H), 1.87 (s, 3H), 2.07 (m, 2H). 2.40 (m, 1H), 3.17 (m, 1H), 3.50 (m, 1H), 4.34 (m, 1H), 4.94 (m, 1H), 5.23 (m, 1 H), 5.34 (m, 1 H), 5.53 (m, 1H), 5.58 (m, 1H), 8.24 (d, 1H), 9.25 (br s, 2H). MS: (M+H) = 281. -317- WO 99/54299 PCT/US99/07945 Example 122 (±)-(2R,3S,5R, 1 'S)-2-(1-Acetamido-4-hydroxy-2-(cis and trans)buten-1 -yl)-3-(cis propen-1 -ylv)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OtBu TBSO Oc t 122A (±)-(2R,3S,5R,1'S)-l-t-Butoxycarbonyl-2-(1 -Acetamido-4-(t butyldimethylsilyloxy)-2-(cis and trans)buten-1 -vyl)-3-(cis-propen-1 -yl)-pyrrolidine 5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example substituting (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido-2 formyl)methyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-l1-t-butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3 formyl-pyrrolidine-5-carboxylic acid t-butyl ester and 4-(t-butyldimethylsilyloxy) butyltriphenylphosphonium bromide for methyltriphenylphosphonium bromide (yield: 23.1 mg, 66.9%). MS: (M+H) = 567.
CH
3 AcHN, OH HO 0H TFA 122B (±)-(2R,3S,5R,1'S)-2-(1-Acetamido-4-hydroxy-2-(cis and trans)buten-1 -yl) 3-(cis-propen--ylv)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'S)-1-t-butoxycarbonyl-2-(1-acetamido -318- WO 99/54299 PCT/US99/07945 2-(cis and trans)-4-hydroxy-butenyl-2-yl)-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 16.9 mg, >100%). 1 H NMR (DMSO-d6): 51.67 (dd, 3H), 1.78 (dt, 1H), 1.91 (s, 3H), 2.44 (m, 1H), 2.50 (m, 1H), 2.56 (m, 1H), 2.65 (m, 1H), 3.23 (m, 1H), 3.54 (m, 1H), 4.40 (m, 1H), 4.47 (m, 2H), 5.01 (m, 1H), 5.26 (m, 1H), 5.54 (m, 2H), 5.63 (m, 1H), 8.32 (d, 1H), 9.27 (br s, 2H). MS: (M+H) = 297. Example 123 (±)-(2R,3S,5R, 1 'S)-2-(1-Acetamido)butyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloride TBDPSO-, OtBu rI N if 0 Ph 123A (±)-(2R,3R,5R)-1l-Benzyl-2-vinyl-3-t-butyldiphenylsilyloxymethyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R, 3R,5R)-1 -Benzyl-2-vinyl-3-hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (30.8 g, 97.1 mmol) was reacted with t-butyldiphenylsilyl chloride (49.5 mL, 190.4 mmol) and imidazole in dichloromethane (650 mL) at 0 oC for 1 hour. The reaction was quenched methanol followed by extraction with dichloromethane (600 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column -319- WO 99/54299 PCT/US99/07945 chromatography on silica gel using 2/1: chloroform/hexane to provide the title compound (yield: 52.9 g, 98%). 1 H NMR (CDCI 3 ) 7.62-7.67 (m, 4H), 7.32-7.44 (m, 6H), 7.25-7.30 (m, 5H), 5.58-5.72 (m, 1H), 5.06-5.14 (m, 2H), 3.90 (d, 1H), 3.72-3.78 (m, 1H), 3.58-3.68 (m, 2H), 3.44-3.52 (m, 2H), 2.26-2.40 (m, 1H), 2.10-2.23 (m, 1H), 1.68-1.78 (m, 1H), 1.38 (s, 9H), 1.03 (s, 9H). MS: (M+H) =556 TBDPSO-, HO OtBu HO Ph 123B (±)-(2R,3R,5R, 1 'RS)-1l-Benzyl-2-(1,2-dihydroxy)ethyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R)-l1-Benzyl-2-vinyl-3-t-butyldiphenylsilyloxymethyl pyrrolidine-5-carboxylic acid t-butyl ester (22.7 g, 41 mmol) was reacted with OsO4 (4%) (2.5 mnL, 0.7 mol.%) and N-methyl morpholine N-oxide (18.5 g, 2.77 eq.) in acetone (500 mL) and water (60 mL) for 48h at room temperature. The reaction was quenched with 10% aqueous Na 2
S
2 0 3 (200 mL). The reaction was concentrated in vacuo and the residue was partitioned between ethyl acetate/water. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 35% ethyl acetate/hexane to provide the title compound (yield: 11 g, 55%). 1 H NMR (DMSO-d 6 ) 6 7.58-7.63 (m, 5H), 7.40-7.48 (m, 7H), 7.20-7.35 (m, 3H), 4.41-4.45 (m, 2H), 3.98 (d, 1H), 3.75-3.84 (m, 2H), 3.50-3.68 (m, 2H), 3.4 3.46 (m, 1H), 3.16-3.25 (m, 1H), 2.97-3.0 (m, 1H), 2.09-2.28 (m, 1H), 1.62-1.89 (m, 1H), 1.34-1.39 (m, 1H), 1.30 (s, 9H), .98,.96 (2s, 9H). -320- WO 99/54299 PCT/US99/07945 MS: (M+H)+=590 TBDPSO, HO '- OtBu N HO 123C (±)-(2R,3R,5R,1'RS)-2-(1,2-dihydroxy)ethyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R, 1'RS)-1 -Benzyl-2-(1,2-dihydroxy)ethyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (11 g, 18.7 mmol) was reacted under 1 atmosphere of hydrogen with 20% Pd(OH) 2 /C (5 g) and in ethanol (40 mL) vigorously stirred for 2.5 days at room temperature. The reaction was filtered, and the catalyst was washed with methanol (3x30 mL). The filtrate was evaporated in vacuo to give the title compound as an oil (yield: 8 g, 94%) TBDPSO-, HO OtBu H BocO HO 123D (±)-(2R. 3R,5R, 1 'R)-1 -t-Butoxvcarbonyl-2-(1,2-dihydroxy)ethyl-3-t butyldiphenyvlsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 40D, substituting (±)-(2R,3R,5R, 1 'RS)-2-(1,2-dihydroxy)ethyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (2R,3R,5R, 1 'RS)-2-(1,2-dihydroxy)ethyl-3-acetoxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester. The residue was purified by column chromatography on silica gel using 35% ethyl acetate/hexane to provide the title compound (yield: 20.5 g, 60%). -321- WO 99/54299 PCT/US99/07945 1 H NMR (DMSO-d 6 ) 7.57-7.60 (m, 4H), 7.38-7.48 (m, 6H), 4.85,4.77 (2d, 1H), 4.45-4.50 (m, 1H), 4.02-4.10 (m, 1H), 3.80-3.95 (m, 1H), 3.73,3.68 (2s, 1H), 3.45-3.67 (m, 2H),3.18-3.28 (m, 2H), 2.36-2.46 (m, 2H), 1,86,1.70 (2d, 1H), 1.40,1.35 (2s, 9H),1.32,1.26 (2s, 9H), 1.0,0.98 (2s, 9H). MS: (M+H) 4 = 600 TBDPSO-, MsO.. OtBu AcO 123E ()-(2R.3R.5R, 1 'R)-1l-t-Butoxycarbonyl-2-(1-methanesulfonvioxy- 2 acetoxy)ethyl-3-t-butyldiphenylsilyvloxvmethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'R)- 1-t-Butoxycarbonyl-2-(1,2-dihydroxy)ethyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (20.5 g, 34.2 mmole) was reacted with acetic anhydride (16.1 mL, 171 mmole) and triethylamine (47.7 mL, 342 mmole) in dichloromethane (360 mL) at 0 0 C for 16h. The reaction was treated with methanol (35 mL) for 10 minutes and diluted with dichloromethane (1300 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was reacted with methanesulfonyl chloride (4.0 mL, 51.3 mmole) and triethylamine (14.3 mL, 103 mmole) in dichloromethane (350 mL) at 0 0 C for 1.5 hours. The reaction was quenched with water (300 mL) and diluted with dichloromethane (1200 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 30% ethyl acetate/hexanes to provide the title compound (yield: 23.8 g, 97%). -322- WO 99/54299 PCT/US99/07945 1 H NMR (DMSO-d 6 ) 67.58-7.62 (m, 4H), 7.38-7.50 (m, 6H), 5.12-5.26 (2m, 1H), 4.06-4.25 (m, 3H), 4.00 (d, 1H), 3.46-3.68 (m, 2H), 3.20,3.18 (2s, 3H), 2.40 2.48 (m, 1H), 2.02,1.99, (2s, 3H), 1.68-1.88 (m, 1H), 1.42,1.36 (2s, 9H), 1.31,1.25 (2s, 9H), 1.00,0.98 (2s, 9H). MS: (M+H) = 720, (M+NH 4 ) =737 TBDPSO-' O OBu 0 O 123F (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-oxiranyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-methanesulfonyloxy-2 acetoxy)ethyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (23.8 g, 33.1 mmole) was reacted with potassium carbonate (10.1 g,66.2 mmole) in methanol (160 mL) and THF (160 mL) at 25 OC for 18 hours. The reaction was concentrated in vacuo. The residue was dissolved in ethyl acetate and washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 25% ethyl acetate/hexanes to provide the title compound, as an oil (yield: 16.7 g, 87%). 1 H NMR (CDCI 3 ) 6 7.60-7.68 (m, 4H), 7.32-7.45 (m, 6H), 4.02-4.28 (m, 2H), 3.67-3.78 (m, 1H), 3.52-3.62 (m, 1H), 3.0-3.08 (m, 1H), 2.68-2.75 (m, 1H), 2.47-2.52 (m, 3H), 1.80-1.90 (m, 1H), 1.48,1.42 (2s, 9H), 1.37,1.35 (2s, 9H), 1.07,1.03 (2s, 9H). MS: (M+H)*= 582 -323- WO 99/54299 PCT/US99/07945
HO
< oc OtBu 0 123G (±)-(2R,3R,5R, 1 'S)-1l-t-Butoxycarbonyl-2-oxiranyl-3-hydroxymethyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R, 1 'S)-1 -t-Butoxycarbonyl-2-oxiranyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (4.17 g, 7.2 mmole) was reacted with tetrabutylammonium fluoride (1M) (14 mL, 14.0 mmole) in THF (7 mL) for 20 minutes at 0 0 C then for 1.5 hours at 250C. The reaction was concentrated in vacuo the residue was dissolved in ethyl acetate and washed with pH 7.0 buffer and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 50% ethyl acetate/hexanes to provide the title compound, as an oil (yield: 2.4 g, 97%). 1 H NMR (DMSO-d 6 ) 5 4.72-4.78 (m, 1H), 3.94-4.05 (m, 2H), 3.35-3.47 (m, 1H), 3.18-3.28 (m, 1H), 3.03-3.08 (m, 1H), 2.63-2.73 (m, 1H), 2.37-2.44 (m, 1H), 2.30-2.36 (m, 1H), 2.08-2.20 (m, 1H), 1.58-1.75 (m, 1H), 1.40 (s, 9H), 1.37,1.34 (2s, 9H). MS: (M+H) = 344, (M+Na) = 366 -324- WO 99/54299 PCT/US99/07945 H o-o OtBu 0 B 0 123H (±)-(2R, 3R,5R,1'S)-l-t-Butoxycarbonyl-2-oxiranyl-3-formyl-pyrrolidine-5 carboxylic Acid t-Butyl Ester. (±)-(2R,3R,5R, 1'S)-1 -t-Butoxycarbonyl-2-oxiranyl-3-hydroxymethyl pyrrolidine-5-carboxylic acid t-butyl ester (2.4 g, 7.0 mmole) and triethylamine (3.9 mL 28.0 mmole) in dichloromethane (70 mL) at 000C was reacted with sulfur trioxide pyridine complex (3.35 g, 21.0 mmole) in dimethylsulfoxide (21 mL) by dropwise addition followed by reaction for an additional 3 hours. The reaction was quenched with water (50 mL) and diluted with ethyl acetate (200 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo to provide the title compound (yield: 2.2 g,). 1 H NMR (DMSO-d 6 ) (rotamers) 5 9.58 and 9.56 (2s, 1H), 4.70 and 4.53 (2m, 1H), 3.96 (dd, J=1.4, 9.2 Hz, 1H), 3.25-3.20 (m, 1H), 2.91 (m, 1H), 2.71 (m, 1H), 2.50-2.28 (m, 3H), 1.42, 1.37, 1.34, and 1.30 (4s, 18H) MS: (M-H)-= 340 B"OtBu B 0 1231 (±)-(2R.3S,5R,1'S)-l-t-Butoxycarbonyl-2-oxiranyl-3-vinyl-pyrrolidine-5 carboxvlic Acid t-Butyl Ester. Triphenylphosphoranylidenemethyl ylide (17.6 mmole) prepared by reacting methyltriphenylphosphonium bromide (12.63 g, 35.4 mmole) and potassium tert-butoxide (1M) (17.6 mL, 17.6 mmole) in THF (70 mL) for 1 hour at -325- WO 99/54299 PCT/US99/07945 250C. (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-oxiranyl-3-formyl-pyrrolidine-5 carboxylic acid t-butyl ester (2.2 g, 6.5 mmole) in THF (10 mL) was added to the above solution at 00C and stirred for 0.5 hours. The reaction was quenched with saturated ammonium chloride (50 mL) and diluted with ethyl acetate (200 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 10% ethyl acetate/hexanes to provide the title compound (yield: 2 g, 84%). 1 H NMR (DMSO-d 6 ) 5 5.80-5.95 (m, 1H), 5.08 (d, 1H), 4.94-5.04 (1H), 4.00-4.07 (m, 1H), 3.59,3.90 (2t, 1H), 3.07-3.16 (m, 1H), 2.73-2.81 (m, 1H), 2.65 2.72 (m, 1H), 2.35-2.48 (m, 1H), 1.59-1.76 (m, 1H), 1.42 (s, 9H), 1.38,1.35 (2s, 9H). MS: (M+H) = 340 MsO OtBu
N
3 123J (±)-(2R,3S,5R,1'R)-l-t-Butoxycarbonyl-2-(1-methanesulfonyloxy-3 azido)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R,1'S)-1l-t-Butoxycarbonyl-2-oxiranyl-3-vinyl-pyrrolidine-5 carboxylic acid t-butyl ester (1.72 g, 5.1 mmole) and ammonium chloride (1.36 g, 25.4 mmole) in ethanol (45 mL) and water (5 mL) was reacted with lithium azide (1.2 g, 24.5 mmole) for 7 hours at 500C. The reaction was concentrated in vacuo and diluted with ethyl acetate (200 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue (2.15 g) was dissolved in dichloromethane (50 mL) and reacted with -326- WO 99/54299 PCT/US99/07945 methanesulfonyl chloride (0.8 mL, 10.2 mmole) and triethylamine (2.8 mL, 20.4 mmole) for 0.5 hours at 0OC. The reaction was quenched with aqueous sodium bicarbonate (50 mL) and diluted with ethyl acetate (200 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 10% ethyl acetate/hexanes to provide the title compound (yield: 1.87 g, 80%). 1 H NMR (DMSO-d 6 ) 6 5.77-5.98 (m, 1H), 4.94-5.11 (m, 3H), 4.12-4.19 (m, 1H), 3.99-4.06 (m, 1H), 3.66,3.71 (2d, 1H), 3.25,3.22 (2s, 3H), 2.92-3.02 (m, 1H), 2.55-2.63 (m, 1H), 1.68-1.82 (m. 1H), 1.45,1.42 (2s, 9H), 1.38,1.36 (2s, 9H). MS: (M+H)*= 461 = HN OtBu B 0 123K (±)-(2R,3S,5R, l1'S)-1 -t-Butoxycarbonyl-2-aziridinyl-3-vinyl-pyrrolidine-5 carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1'S)-1 -t-Butoxycarbonyl-2-(1-methanesulfonyloxy-3 azido)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (2.12 g, 4.6 mmole) was reacted with triphenylphosphine (1.81 g, 6.9 mmole) in THF (30 mL) and water (7.5 mL) at 65 0 C for 1 hour. The reaction was concentrated in vacuo and redissolved in ethyl acetate (200 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 4% methanol in dichloromethane to provide 2 g of the crude title compound containing approximately 60% product and 40% Ph 3 PO which was used directly for acylation. -327- WO 99/54299 PCT/US99/07945 1 H NMR (DMSO-d 6 ) 6 5.78-5.5.98 (m, 1H), 4.12 (d, 1H), 3.42,3.19 (2d, 1H), 2.53-2.73 (m, 2H), 2.00-2.15 (m, 1H), 1.68-1.76 (m, 1H), 1.62-1.68 (m, 1H), 1.41 (s, 9H), 1.37,1.36 (2s, 9H). MS: (M+H) = 339, (M+Na) = 361 = AcN OtBu AcNT OC B 0 123L (±)-(2R,3S,5R, l'S)-1 -t-Butoxycarbonyl-2-(N-acetylaziridinyl)-3-vinyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R, 3S,5R, 1 'S)-1 -t-Butoxycarbonyl-2-aziridinyl-3-vinyl-pyrrolidine-5 carboxylic acid t-butyl ester (1.03 g, 3.1 mmole) was reacted with acetic anhydride (.42 mL, 4.7 mmole) and triethylamine (1.3 mL, 9.3 mmole) in dichloromethane (30 mL) at 25 0 C for I hours. The reaction was quenched with water (50 mL) and diluted with ethyl acetate (200 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 20% ethyl acetate/hexanes to provide the title compound (yield: .75 g, 64%). 1 H NMR (DMSO-d 6 ) 6 5.78-5.98 (m, 1H), 5.05 (d, 1H), 4.98,4.94 (2d, 1H), 4.12-4.20 (m, 1H), 3.54,3.42 (2dd, 1H), 2.54-2.98 (m, 3H), 2.40,2.49 (2d, 1H), 2.15,2.19 (2d, 1H), 2.02,2.04 (2s, 3H), 1.68-1.82 (m, 1H), 1.42 (s, 9H), 1.48.1.45 (2s, 9H). MS: (2M+Na)*= 783 -328- WO 99/54299 PCT/US99/07945 = AcN. OtBu 'N H Boc 123M (±)-(2R,3S,5R, 1 'S)-1l-t-Butoxycarbonyl-2-(1-acetamido)butyl-3-vinyl pyrrolidine-5-carboxylic Acid t-Butyl Ester. To a suspension of copper(I) bromide-dimethyl sulfide complex (0.051g, 0.248 mmol) in THF (1.0 ml) at 0 oC was added ethylmagnesium bromide (1M) (1.0 ml, 1.0 mmol) in THF. After stirring for 10 minutes at 0 oC, a portion of this solution (0.60 ml) was added dropwise to a solution of (±)-(2R,3S,5R,1'S)-1-t Butoxycarbonyl-2-(N-acetylaziridinyl)-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (0.020 g, 0.053 mmole) in THF (0.40 ml) at -78 0 C. After stirring for 20 minutes at -780C, the reaction was warmed to 0 oC and stirred for 30 minutes. The reaction was quenched with saturated ammonium chloride (1.0 mL) and diluted with ethyl acetate (10 mL). The organic layer was washed with water and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using a gradient of 0-75% ethyl acetate/hexanes to provide the title compound (yield: 0.004 g, 19%). 1 H NMR (DMSO-d 6 ) (rotamers) 5 7.48 (d, J=9.5Hz, 1H), 5.98-5.80 (m, 1H), 5.00-4.90 (m, 2H), 4.45-4.25 (m, 1H), 3.96-3.91 (m, 1H), 3.60-3.57 and 3.53-3.50 (2m, 1H), 2.91-2.76 (m, 1H), 2.59-2.42 (m, 1H), 1.80 (s, 3H), 1.73-1.59 (m, 1H), 1.42 and 1.41 (2s, 9H), 1.40-1.15 (m, 4H), 1.37 and 1.34 (2s, 9H), 0.89-0.82 (m, 3H) MS: (M-H) = 409, (M+H) = 411 -329- WO 99/54299 PCT/US99/07945 AcHN ,OH A H 7 PH 0 HCI 123N (±)-(2R,3S,5R, 1 'S)-2-(1-Acetamido)butyl-3-vinyl-pyrrolidine-5-carboxvlic Acid Hydrochloric Acid Salt The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3S,5R,1'S)-1l-t-butoxycarbonyl-2-(1 acetamido)butyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (2R,3R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 3.1 mg, 99%). 1 H NMR (DMSO-d 6 ) 5 8.11 (d, J=7.3Hz, 1H), 5.76-5.69 (m, 1H), 5.16 (d, J=17.1Hz, 1H), 5.07 (dd, J=1.5, 10.3Hz, 1H), 4.30 (dd, J=7.3, 9.8Hz, 1H), 4.13 (m, 1H), 3.50 (dd, J=5.9, 9.8Hz, 1H), 2.90 (m, 1H), 2.39 (m, 1H), 1.92-1.85 (m, 1H), 1.87 (s, 3H), 1.52-1.18 (m, 4H), 0.85 (t, J=7.3, 3H) MS: (M-H)- = 253, (M+H)* = 255 Examples 124-130 1. Organocuprate AcHN. OH OtBu AcHN. OH AcN N 2. 6N HCI HN O R O HCI The following title compounds were prepared in two steps according to the methods described in Examples 123M and 123N, the denoted reagents and their -330- WO 99/54299 PCT/US99/07945 respective methods of preparation are substituted in place of diethylcuprate and its preparation in Example 123M for step 1. Example 124 AcHN, OH H0 HCI (±)-(2R,3S,5R, 1 'S)-2-(1-Acetamido)hexyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloric Acid Salt The organocuprate reagent was prepared from a Grignard reagent and a catalytic quantity of copper(l) bromide-dimethyl sulfide complex according to the methods described in Example 123M, substituting 2M butylmagnesium chloride for 1M ethylmagnesium bromide. 1 H NMR (MeOD-d 3 ) 5 5.82-5.70 (m, 1H), 5.29 (d, J=17.0Hz, 1H), 5.17 (dd, J=1.3, 10.2Hz, 1H), 4.35 (dd, J=7.5, 10.2Hz, 1H), 4.19 (m, 1H), 3.65 (dd, J=3.4, 9.8Hz, 1H), 3.01 (m, 1H), 2.55 (m, 1H), 2.08-1.97 (m, 1H), 2.04 (s, 3H), 1.62-1.31 (m, 8H), 0.91 (t, J=6.4Hz, 3H) MS: (M-H) = 281, (M+H) = 283 -331- WO 99/54299 PCT/US99/07945 Example 125 AcHN H OH N H0 HCI (+)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-4-methyl)pentyl-3-vinyl-pyrrolidine-5 carboxylic Acid Hydrochloric Acid Salt The organocuprate reagent was prepared from a Grignard reagent and a catalytic quantity of copper(I) bromide-dimethyl sulfide complex according to the methods described in Example 123M, substituting iso-butylmagnesium chloride for ethylmagnesium bromide.. 1 H NMR (MeOD-d 3 ) 8 5.83-5.71 (m, 1H), 5.29 (dd, J=0.7,17.0Hz, 1H), 5.17 (dd, J=0.7, 10.2Hz, 1H), 4.34 (dd, J=7.5, 10.2Hz, 1H), 4.15 (m, 1H), 3.66 (dd, J=3.4, 9.8Hz, 1H), 3.01 (m, 1H), 2.55 (m, 1H), 2.08-1.97 (m, 1H), 2.04 (s, 3H), 1.65-1.10 (m, 5H), 0.91 (d, J=6.4Hz, 3H), 0.91 (d, J=6.5Hz, 3H) (M+H)* = 283 -332- WO 99/54299 PCT/US99/07945 Example 126 = AcHN OH N H 0 HCI (±)-(2R,3S, 5R, 1 'S)-2-(1 -Acetamido-3,3dimethyl)butyl-3-vinyl-pyrrolidine-5 carboxylic Acid Hydrochloric Acid Salt The organocuprate reagent was prepared from a Grignard reagent and a catalytic quantity of copper(l) bromide-dimethyl sulfide complex according to the methods described in Example 123M, substituting 1M tert-butylmagnesium chloride for 1M ethylmagnesium bromide. 1 H NMR (MeOD-d 3 ) 8 5.84-5.71 (m, 1H), 5.31 (d, J=17.0Hz, 1H), 5.19 (d, J=10.2Hz, 1H), 4.39-4.33 (m, 2H), 3.66 (dd, J=3.4, 9.8Hz, 1H), 3.02 (m, 1H), 2.57 (m, 1H), 2.08-1.97 (m, 1H), 2.02 (s, 3H), 1.55 (dd, J=9.5, 14.6Hz, 1H), 1.42 (dd, J=1.4, 14.6Hz, 1H), 0.95 (s, 9H) (M+H) = 283 -333- WO 99/54299 PCT/US99/07945 Example 127 AcHN OH 'N HH HCI (+)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-phenyl)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloric Acid Salt Lithium diphenylcurpate was prepared according to the method described by Lipshutz, B. H. in Or-qanometallics in Synthesis; Schlosser, M., Ed.; Wiley and Sons: New York, 1994; p.292. This cuprate was used according to the methods described in Example 123M, substituting lithium diphenylcuprate for the Grignard derived diethylcuprate complex. 1 H NMR (MeOD-d 3 ) 7.35-7.21 (m, 5H), 5.87-5.75 (m, 1H), 5.37 (d, J=16.6Hz, 1H), 5.26 (dd, J=1.0, 10.2Hz, 1H), 4.53 (m, 1H), 4.37 (dd, J=7.5, 9.8Hz, 1H), 3.70 (dd, J=3.7, 9.8Hz, 1H), 3.11 (m, 1H), 2.97 (dd, J=6.1, 14.2Hz, 1H), 2.84 (dd, J=9.5, 14.2Hz, 1H), 2.59 (m, 1H), 2.08-1.99 (m, 1H), 1.93 (s, 3H) (M-H)-= 301, (M+H) = 303 -334- WO 99/54299 PCT/US99/07945 Example 128 = AcHN. OH N N H O HCI (±)-(2R,3S, 5R, 1 'S)-2-(1 -Acetamido-4-phenyl)butyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloric Acid Salt The organocuprate reagent was prepared from a Grignard reagent and a catalytic quantity of copper(I) bromide-dimethyl sulfide complex according to the methods described in Example 123M, substituting 1M phenethylmagnesium chloride for 1M ethylmagnesium bromide. 1 H NMR (MeOD-d 3 ) 8 7.29-7.13 (m, 5H), 5.77-5.65 (m, 1H), 5.24 (d, J=16.6Hz, 1H), 5.13 (dd, J=1.0, 9.8Hz, 1H), 4.33 (dd, J=7.5, 10.2Hz, 1H), 4.22 (m, 1H), 3.62 (dd, J=3.4, 9.8Hz, 1H), 2.98 (m, 1H), 2.63 (m, 2H), 2.54 (m, 1H), 2.06-1.95 (m, 1H), 2.03 (s, 3H), 1.79-1.55 (m, 4H) (M-H)= 329, (M+H) = 331 -335- WO 99/54299 PCT/US99/07945 Example 129 AcHN- OH 'N HH HCI (±)-(2R,3S, 5R, 1 'S)-2-(1-Acetamido-3-phenyl)butyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloric Acid Salt The organocuprate reagent was prepared from a Grignard reagent and a catalytic quantity of copper(I) bromide-dimethyl sulfide complex according to the methods described in Example 123M, substituting 2M benzylmagnesium chloride for 1M ethylmagnesium bromide. 1 H NMR (MeOD-d 3 ) 8 7.30-7.17 (m, 5H), 5.82-5.70 (m, 1H), 5.28 (d, J=17.0Hz, 1H), 5.17 (d, J=11.2Hz, 1H), 4.33 (dd, J=7.5, 10.2Hz, 1H), 4.18 (m, 1H), 3.64 (dd, J=3.4, 9.8Hz, 1H), 3.01 (m, 1H), 2.78 (m, 1H), 2.66-2.50 (m, 2H), 2.07 (s, 3H), 2.07-1.85 (m, 3H) (M-H)-= 315, (M+H) = 317 -336- WO 99/54299 PCT/US99/07945 Example 130 (±)-(2R,3S, 5R, 1 'S)-2-(1 -Acetamido-2-propen-2-yl)ethyl -3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt BocN O t B u 130A (±)-(2R,3S,5R,1'S)-l-t-Butoxycarbonyl-2-(N-t-Butoxycarbonylaziridinyl)-3 vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1 'S)-I -t-Butoxycarbonyl-2-azirid inyl-3-vinyl-pyrrolid ine-5 carboxylic acid t-butyl ester (0.058 g, 0.17 mmole) was reacted with di-t butyldicarbonate (95 mg, 0.44 mmole) and triethylamine (0.12 mL, 0.86 mmole) in dichloromethane (2.0 mL) at room temperature for 1 hour. The reaction was quenched with saturated sodium bicarbonate (1.0 mL) and diluted with ethyl acetate (20 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using a gradient of 0-15% ethyl acetate/dichloromethane to provide the title compound (yield: 0.060 g, 80%). 1 H NMR (DMSO-d 6 ) (rotamers) 8 5.97-5.78 (m, 1H), 5.06-4.93 (m, 2H), 4.15 (dd, J=2.0, 9.8Hz, 1H), 3.40-3.28 (m, 1H), 2.94-2.49 (m, 3H), 2.39 and 2.33 (2d, J=6.1, 6.4Hz, 1H), 2.17 and 2.11 (2d, J=3.7, 3.4, 1H), 1.81-1.69 (m, 1H), 1.42-1.36 (m, 27H) MS: (M+Na) = 461 (weak) -337- WO 99/54299 PCT/US99/07945 BocHN, OtBu N H BocO O 130B (±)-(2R,3S,5R, 1 'S)-1l-t-Butoxycarbonyl-2-(1-N-t-butoxycarbonylamino-2 propen-2-yl)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. To a suspension of copper(l) bromide-dimethyl sulfide complex (0.026g, 0.127 mmol) in THF (1.0 ml) at 0 oC was added isopropenylmagnesium bromide (0.5M) (1.0 ml, 0.50 mmol) in THF. After stirring for 10 minutes at 0 oC, the mixture was cooled to -78 oC and a solution of (±)-(2R,3S,5R,1'S)-1-t butoxycarbonyl-2-(N-t-butoxycarbonylaziridinyl)-3-vinyl-pyrrolid idine-5-carboxyl ic acid t-butyl ester (0.030 g, 0.068 mmole) in THF (1.0 ml) was added dropwise. After stirring for 10 minutes at -780C, the reaction was warmed to 0 oC and stirred for 2 hours. The reaction was quenched with saturated ammonium chloride (1.0 mL) and diluted with ethyl acetate (10 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using a gradient of 0-10% ethyl acetate/dichloromethane to provide the title compound (yield: 0.026 g, 79%). 1 H NMR (DMSO-d 6 ) (rotamers) 8 6.64 (m, 1H), 5.96-5.76 (m, 1H), 4.98 4.89 (m, 2H), 4.76-4.68 (m, 2H), 4.40-4.25 (m, 1H), 3.94 (m, 1H), 3.60-3.53 (m, 1H), 3.02-2.86 (m, 1H), 2.62-2.42 (m, 1H), 2.10-1.99 (m, 2H), 1.72 and 1.70 (2s, 3H), 1.72-1.55 (m, 1 H), 1.44-1.34 (m, 27H) MS: (M-H)-= 479, (M+H)* = 481 -338- WO 99/54299 PCT/US99/07945 Ac BocN. OtBu N H BocO 130C (±)-(2R,3S. 5R,1'S)-1l-t-Butoxycarbonyl-2-(1-N-t-butoxycarbonylacetamido 2-propen-2-yl)ethyl -3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-N-t-butoxycarbonylamino-2 propen-2-yl)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (0.024 g, 0.050 mmole) was reacted with lithium hexamethyldisilazide (1 M) (0.60 mL, 0.60 mmole) in THF (2.0 mL) at -25 0 C for 1 hour. To the above reaction was then added acetyl chloride (0.085 mL, 1.20 mmole) at -250C and the mixture was stirred for 30 minutes. The reaction was quenched with saturated sodium bicarbonate (2.0 mL) and stirred for 30 minutes at room temperature. The reaction was diluted with ethyl acetate (20 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using a gradient of 0-15% ethyl acetate/hexanes to provide the title compound (yield: 0.015 g, 58%) along with unreacted starting material. 1 H NMR (DMSO-d 6 ) (rotamers) 8 6.01-5.84 (m, 1H), 4.99-4.89 (m, 2H), 4.76-4.58 (m, 3H), 4.33 and 4.23 (2d, J=7.8, 8.1Hz, 1H), 4.13-4.04 (m, 1H), 2.69 (m, 1H), 2.62-2.42 (m, 1H), 2.29 (br s, 3H), 2.35-2.14 (m, 2H), 1.76-1.55 (m, 1H), 1.60 (s, 3H), 1.50-1.35 (m, 27H) MS: (M+H)
+
= 523 -339- WO 99/54299 PCT/US99/07945 = AcHN OH 'N HHO 0 TFA 130D (+)-(2R,3S,5R, 1 'S)-2-(1-Acetamido-2-propen-2-yl)ethyl -3-vinyl-pyrrolidine 5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R,1 'S)-l-t-butoxycarbonyl-2-(1-N-t butoxycarbonylacetamido-2-propen-2-yl)ethyl -3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. (yield: 12 mg, 99%). 1 H NMR (MeOD-d 3 ) 6 5.83-5.70 (m, 1H), 5.30 (dd, J=0.7, 17.0Hz, 1H), 5.19 (d, J=10.2Hz, 1H), 4.79 (s, 1H), 4.71 (s, 1H), 4.46 (m, 1H), 4.30 (dd, J=7.8, 9.8Hz, 1H), 3.66 (dd, J=3.7, 9.8Hz, 1H), 3.03 (m, 1H), 2.56 (m, 1H), 2.40-2.19 (m, 2H), 2.08-1.96 (m, 1H), 2.01 (s, 3H), 1.76 (s, 3H) (M-H)-= 265, (M+H) = 267 Examples 131-135 1. Organocuprate OtBu , AcHN. OH BocHN NO 2. LiHMDS H BoH H 3. AcCI 0 4. TFA/CH 2
CI
2 R TFA The following title compounds were prepared in 4 steps according to the methods described in Example 130 the denoted reagents for step 1 and their -340- WO 99/54299 PCT/US99/07945 respective methods of preparation are substituted in place of isopropenyl cuprate and its preparation in 130B Example 131 = AcHN OH 'N K H0 TFA (+)-(2R.,3S,5R. 1 'S)-2-(1-Acetamido-1 -(cis and trans)-propen-1-yl)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The organocuprate reagent was prepared from a Grignard reagent and a catalytic quantity of copper(I) bromide-dimethyl sulfide complex according to the methods described in Example 130B, substituting 0.5M 1-propenylmagnesium bromide (mixture of cis and trans isomers) for 0.5M isopropenylmagnesium bromide. 1 H NMR (MeOD-d 3 ) (2:1 trans:cis ratio) 6 5.81-5.54 (m, 2H), 5.43-5.30 (m, 1H), 5.33-5.27 (m, 0.33H, cis isomer), 5.31-5.25 (m, 0.66H, trans isomer), 5.20 5.15 (m, 1H), 4.26-4.17 (m, 2H), 3.65 (dd, J=3.4, 9.8Hz, 1H), 2.98 (m, 1H), 2.58 2.48 (m, 1H), 2.45-2.19 (m, 2H), 2.08-1.94 (m, 1H), 2.02 (s, 3H), 1.68 (m, 2H, trans isomer), 1.63 (m, 1H, cis isomer) (M-H)-= 265, (M+H) = 267 -341- WO 99/54299 PCT/US99/07945 Example 132 AcHN OH . HH HH0 TFA (+)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-allyl)methyl-3-vinyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The organocuprate reagent was prepared from a Grignard reagent and a catalytic quantity of copper(l) bromide-dimethyl sulfide complex according to the methods described in Example 130B, substituting 1M vinylmagnesium bromide for 0.5M isopropenylmagnesium bromide. 1 H NMR (MeOD-d 3 ) 85.83-5.70 (m, 2H), 5.28 (d, J=17.0Hz, 1H), 5.19-5.13 (m, 3H), 4.28 (m, 1H), 4.19 (dd, J=8.5, 9.1Hz, 1H), 3.66 (dd, J=3.4, 9.5Hz, 1H), 2.99 (m, 1H), 2.57-2.48 (m, 1H), 2.44-2.26 (m, 2H), 2.05-1.93 (m, 1H), 2.01 (s, 3H) (M+H) = 253 -342- WO 99/54299 PCT/US99/07945 Example 133 AcHN. OH N H 0 TFA (+)-(2R,3S,5R,1 'S)-2-(1 -Acetamido)-2-(1 -buten-2-yl)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt The organocuprate reagent was prepared from a Grignard reagent and a catalytic quantity of copper(I) bromide-dimethyl sulfide complex according to the methods described in Example 130B, substituting 0.5M 1-buten-2-ylmagnesium bromide for 0.5M isopropenylmagnesium bromide. 1 H NMR (MeOD-d 3 ) 5 5.81-5.73 (m, 1H), 5.30 (d, J=17.1Hz, 1H), 5.19 (d, J=10.0Hz, 1H), 4.93 (s, 1H), 4.83 (s, 1H), 4.45 (m, 1H), 4.31 (dd, J=7.6, 9.8Hz, 1H), 3.69 (dd, J=3.2, 9.8Hz, 1H), 3.03 (m, 1H), 2.59-2.53 (m, 1H), 2.38 (dd, J=5.9, 14.9Hz, 1H), 2.30 (dd, J=9.5, 14.9Hz, 1H), 2.07 (q, J=7.6Hz, 2H), 2.05 1.99 (m, 1H), 2.01 (s, 3H), 1.05 (t, J=7.6Hz, 3H)
(M+H)
+ = 281 -343- WO 99/54299 PCT/US99/07945 Example 134 AcHN. N OH
H
3 C H
CH
3 TFA (+)-(2R,3S,5R,1'S)-2-(1-Acetamido-2-(trans-2-buten-2-yl)ethyl-3-vinyl-pyrrolidine 5-carboxylic Acid Trifluoroacetic Acid Salt The organocuprate reagent was prepared from a Grignard reagent and a catalytic quantity of copper(l) bromide-dimethyl sulfide complex according to the methods described in Example 130B, substituting 0.5M 1-methyl-1 propenylmagnesium bromide for 0.5M isopropenylmagnesium bromide. 1 H NMR (MeOD-d 3 ) 6 5.83-5.71 (m, 1H), 5.41 (q, J=6.8Hz, 1H), 5.31 (d, J=17.3Hz, 1H), 5.19 (d, J=10.2Hz, 1H), 4.42 (m, 1H), 4.31 (dd, J=7.5, 9.8Hz, 1H), 3.61 (dd, J=4.0, 9.8Hz, 1H), 3.01 (m, 1H), 2.62-2.52 (m, 1H), 2.46 (dd, J=9.5, 13.9Hz, 1H), 2.26 (dd, J=5.8, 13.9Hz, 1H), 2.09-1.99 (m, 1H), 2.00 (s, 3H), 1.72 (s, 3H), 1.59 (d, J=6.8Hz, 3H) (M+H) = 281 -344- WO 99/54299 PCT/US99/07945 Example 135 AcHN. OH N
-
H 0 TFA (+)-(2R,3S.5R,1'S,3'RS)-2-(1-Acetamido-3-methyl)pentvl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt The organocuprate reagent was prepared from a Grignard reagent and a catalytic quantity of copper(l) bromide-dimethyl sulfide complex according to the methods described in Example 130B, substituting 2M sec-butylmagnesium bromide for 0.5M isopropenylmagnesium bromide. 1 H NMR (MeOD-d 3 ) (1:1 mixture of methyl isomers) 8 5.82-5.69 (m, 1H), 5.27 (d, J=17.0Hz, 0.5H), 5.25 (d, J=17.0Hz, 0.5H), 5.15 (d, J=10.2Hz, 1H), 4.33 (m, 1H), 4.18 (dd, J=2.7, 7.5Hz, 0.5H), 4.15 (dd, J=3.0, 7.8Hz, 0.5H), 3.62 (dd, J=3.1, 9.8Hz, 0.5H), 3.57 (dd, J=4.07, 9.8Hz, 0.5H), 2.97 (m, IH), 2.57-2.47 (m, 1H), 2.03-1.92 (m, 1H), 2.03 (s, 1.5H), 2.02 (s, 1.5H), 1.72-1.06 (m, 5H), 0.95 0.86 (m, 6H) (M+H) = 283 -345- WO 99/54299 PCT/US99/07945 Example 136 (±)-(2R,3S, 5R.1'RS)-2-(1 -Acetamido-1-(N-methyl-N-benzylcarbamoyl)methyl-3 vinyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt AcHN, OtBu N HO BO 136A (±)-(2R.3S,5R, 1'R)-1l-t-Butoxvcarbonyl-2-(1-acetamido-1l-carboxyl)methyl 3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method of Example 2B substituting (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1-acetamido-1 formyl)methyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester for (±) (2R,3R,5R, 1 'S)-1 -benzyl-2-( 1 -acetamido-3-ethyl)pentyl-3-formyl-pyrrolidine-5 carboxylic acid t-butyl ester. AcHN OtBu Ph 0NCf pBOC'
CH
3 136B (±)-(2R,3S,5R, 1 'RS)-1 -t-Butoxycarbonyl-2-(1-acetamido-2-(N-methyl-N benzvylcarbamoyl)methyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1 -acetamido-1 -carboxyl)methyl 3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (36 mg, 0.09 mmole) was reacted with N-methyl-N-benzylamine (32 mg, 0.26 mmole), dimethylaminopyridine (1mg, 0.008 mmole) and 1-(3-dimethylaminopropyl)-3 ethylcarbodiimide hydrochloride (30mg, 0.16mmole) in DMF (3 mL) at 250C for 16 hours. The reaction was quenched with water (3 mL) and diluted with ethyl -346- WO 99/54299 PCT/US99/07945 acetate (20mL). The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 50% ethyl acetate/hexanes to provide the title compound. MS: (M+H)+=516, (M-H)- =514 AcHN OH N H H 0
CH
3 N O TFA Ph 136C (±)-(2R,3S,5R, 1 'RS)-2-(1-Acetamido-1l-(N-methyl-N benzylcarbamovyl)methyl-3-vinyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'RS)-1 -t-butoxycarbonyl-2-(1 acetamido-2-(N-methyl-N-benzylcarbamoyl)methyl-3-vinyl-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t butyl ester (yield: 7 mg, 95%). 1 H NMR (DMSO-d 6 ) 8 8.52( d, J=9.7HZ, 1H), 7.30(m, 5H), 5.65(m, 1H), 5.12(m, 4H), 4.62(m, 1H), 4.40(m, 2H), 3.70(m, 1H), 2.90(s, 3H), 2.20(m, 2H), 1.96(s, 3H), MS: (M+H) =360, (M+23) =382 -347- WO 99/54299 PCT/US99/07945 Example 138 (±)-(2R,3S,5R. 1'R)-2-(1 -Acetamido-2-(N-phenyl-carbonyloxy)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt AcHN. OtBu Ph'NN O B0 C Ph, N--0 H 138A (±)-(2R,3S,5R,1'R)-1-t-Butoxycarbonyl-2-(1-acetamido-2-N-phenyl carbonyloxy)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R,1'R)- 1l-t-Butoxycarbonyl-2-( 1-acetamido-2-hydroxy)ethyl-3 vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (18 mg, 0.045 mmole) was reacted with phenylisocyanate (16 mg, 0.14 mmole) and pyridine(0.1 ml) in THF (3 mL) at 25 0 C for 16 hours. The reaction was quenched with water (2 mL) and diluted with ethyl acetate (10 mL). The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 50% ethyl acetate/hexanes to provide the title compound (yield:7.5 mg, 33%). MS: (M+H)*=518, (M-H)- =516 -348- WO 99/54299 PCT/US99/07945 AcHN. OH ' N 0) 0 0 N-Ph TFA H 138B (±)-(2R,3S,5R, 1 'R)-2-(1 -Acetamido-1 -(N-phenylcarbonyloxy)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R)-1 -t-butoxycarbonyl-2-(1 acetamido-2-N-phenyl-carbonyloxy)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1 -t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 4 mg, 95%). 1 H NMR (DMSO-d 6 ) d 8.36( d, J=9.7HZ, 1H), 7.30(m, 5H), 5.78(m, 1H), 5.22(m, 1H), 5.10O(m, 1H), 4.58(m, 1H), 4.45(m, 1H), 4.14(, 2H), 3.58(m, 1H), 2.88( m, 1H), 2.27(m, 1H), 2.12(m, 1H), 1.88(s, 3H) MS: (M+H) =362, (M+23) + =384, (M-H)-=360, (M+35)-=396 -349- WO 99/54299 PCT/US99/07945 Example 139 (±)-(2R,3S,5R,1'R)-2-(1 -Acetamido-1 -isobutyryloxy)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt = AcHN, OtBu N O 0 0 139A (±)-(2R.3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 isobutyryiloxy)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R, 1'R)-l1-t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy)ethyl-3 vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (15 mg, 0.04 mmole) was reacted with isobutyryl chloride (8 mg, 0.08 mmole) and triethylamine (8 mg, 0.08 mmole) in dichloromethane (4 mL) at 0 0 C for 2 hours. The reaction was quenched with water (3 mL) and diluted with ethyl acetate (20 mL). The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 30% ethyl acetate/hexanes to provide the title compound (yield: 11 mg, 63%). MS: (M+H)+=469, (M-H) =467 -350- WO 99/54299 PCT/US99/07945 AcHON OH N I TFA 139B (±)-(2R,3S,5R, 1'R)-2-(1-Acetamido-1 -isobutvryloxv)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-isobutyryloxy)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 6.0 mg, 96%). 1 H NMR (DMSO-d 6 ) 6 8.00( d, J=9.9HZ, 1H), 5.63(m, 1H), 5.08(m, 1H), 4.98(m, 1H), 4.35(m, 1H), 4.25(m, 1H), 4.08(m, 1H), 3.55(m, 1H), 3.45(m, 1H), 3.38(m, 1H), 2.83(m, 1H), 2.33(m, 1H), 1.78(s, 3H) MS: (M+H)* =243, (M+23) =265, (M-H)- =241 -351- WO 99/54299 PCT/US99/07945 Example 140 (±)-(2R,3S,5R, 1 'R)-2-(1 -Acetamido-2-N-ethyl-thiocarbonyloxy)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt AcHN. OtBu HBoc6 OO S N H 140A (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-N-ethyl thiocarbonyloxy)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R,1'R)-I -t-Butoxycarbonyl-2-(1 -acetamido-2-hydroxy)ethyl-3 vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (17 mg, 0.04 mmole) was reacted with ethylisothiocyanate ( 19 mg, 0.21 mmole) in pyridine (2 mL) at 70 0 C for 17 hours. The reaction was quenched with water (3 mL) and diluted with ethyl acetate (20 mL). The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 70% ethyl acetate/hexanes to provide the title compound (yield: 10 mg, 48%). MS: (M+H)*=486, (M+23) =508, (M-H) = 485 -352- WO 99/54299 PCT/US99/07945 AcHON OH 'N 0 H 0 S N TFA H 140B (±)-(2R,3S,5R, 1'R)-2-(1-Acetamido-2-N-ethyl-thiocarbonyloxy)ethyl-3 vinyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R)-1 -t-butoxycarbonyl-2-(1 acetamido-2-N-ethyl-thiocarbonyloxy)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 7 mg, 94%). 1 H NMR (DMSO-d 6 ) 88.30(d, J=9.7HZ, 1H), 5.78 (m, 1H), 5.25(m, 1H), 5.12(m, 1H), 4.50(m, 1H),4.33(m, 1H), 4.18(m, 2H), 3.72(m, 1H), 3.55(m, 2H), 2.30(m, 1H), 2.10(m, 1H), 1.82(s, 3H), 1.17(m, 3H) MS: (M+H) =330, (M-H)- =328 -353- WO 99/54299 PCT/US99/07945 Example 141 (±)-(2R,3S,5R, 1 'S)-2-(1-Acetamido-2-amino)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid HydrochlorideSalt AcHN', N OtBu BocHNO 141A (±)-(2R, 3S,5R,1'S)-l-t-Butoxycarbonyl-2-(1l-acetamido-2-t butoxvcarbonylamino)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1 'S)-1 -t-Butoxycarbonyl-2-(1 -acetamido-2-azido)ethyl-3-vinyl pyrrolidine-5-carboxylic acid t-butyl ester (9.5 mg, 0.022 mmole) was reacted with triphenylphosphine (23.5 mg, 0.090 mmole) in ethanol (180 L) and water (45 pL) at 70 0 C for 30 minutes. The reaction mixture was concentrated in vacuo. The residue was dissolved in dichloromethane (220 pL) and to it was added di-tert butyl dicarbonate (7.3 mg, 0.034 mmol) and N,N-diisopropylethylamine (11.7 mL, 0.067 mmol) at 25 0 C. After 1 hour the reaction mixture was diluted with water and extracted with ethyl acetate. The organic layers were combined, washed with brine, dried over MgSO 4 , and concentrated in vacuo. The residue was purified by chromatography on silica gel using 100% dichloromethane to 50% dichloromethane/ethyl acetate to provide the title compound (yield: 7.5 mg, 67%). 1 H NMR (DMSO-d 6 ) (rotamers) 5 7.51(d, J=10.5Hz, 1H), 6.80-6.66(m, 1H), 5.90-5.76(m, 1H), 5.02-4.90(m, 2H), 4.38-4.19(m, 1H), 3.98-3.94(m, 1H), 3.68 3.62(m, 1H), 3.09-2.73(m, 2H), 2.60-2.42(m, 1H), 1.80(s, 3H), 1.72-1.62(m, 1H), 1.42-1.34(m, 27H). MS: (M+H)+=498, (M+Na)+=520, (M-H)-=496, (M+CI)-=532 -354- WO 99/54299 PCT/US99/07945 = AcHN- OH N H0
H
2 N 2 HCI 141B (±)-(2R. 3S,5R, 1l'S)-2-(1-Acetamido-2-amino)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid DiHydrochloride The title compound was prepared according to the method described in Example 1 K, substituting (±)-(2R,3S,5R, I'S)-1 -t-butoxycarbonyl-2-(1-acetamido 2-t-butoxycarbonylamino)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester (yield: 3.65 mg, 99%). 1 H NMR (DMSO-d 6 ) 5 8.24(d, J=7.9Hz, 1H), 5.75-5.68(m, 1H), 5.16(d, J=17.1Hz, 1H), 5.06(d, J=10.4Hz, 1H), 4.37-4.27(m, 2H), 3.60-3.16(m, 2H), 3.00 2.88(m, 2H), 2.46-2.36(m, 1H), 1.91-1.81(m, 1H), 1.86(s, 3H). MS: (M+H) =242, (M+Na)f=264, (M-H)=240, (2M-H)-=481 -355- WO 99/54299 PCT/US99/07945 Example 142 (±)-(2R,3S,5R,1'S)-2-(1 -Acetamido-2-acetamido)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Hydrochloride = AcH N AcHN BO 142A (±)-(2R.,3S,5R,1 'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-acetamido)ethyl 3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1'S)-1 -t-Butoxycarbonyl-2-(1 -acetamido-2-amino)ethyl-3 vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (9.4 mg, 0.024 mmole) was reacted with acetic anhydride (11.2 pL) and triethylamine (33.1 pL) in dichloromethane (0.23 mL) at 0 0 C for 1 hour. The reaction was diluted with water (3 mL), extracted with ethyl acetate (12 mL), washed with brine, dried over MgSO 4 , and concentrated in vacuo. The residue was purified by chromatography on silica gel using 100% ethyl acetate to 90% ethyl acetate/methanol to provide the title compound (yield: 6.8mg, 66%). 1 H NMR (DMSO-d 6 ) (rotamers) 6 7.79-7.74(m, 1H), 7.54(d, J=9.8Hz, 1H), 5.97-5.81(m, 1H), 5.01-4.91(m, 2H), 4.36-4.27(m, 1H), 3.97-3.90(m, 1H), 3.68 3.63(m, 1H), 3.21-3.15(m, 1H), 3.10-2.76(m, 1H), 2.88-2.78(m, 1H), 2.58-2.45(m, 1H), 1.81(s, 3H), 1.78(s, 3H), 1.76-1.64(m, 1H), 1.42-1.36(m, 18H). MS: (M+H) =439, (M+Na)+=462, (M-H)-=438, (M+35)-=474 -356- WO 99/54299 PCT/US99/07945 AcHN OH AcHN HCI 142B (±)-(2R.3S,5R,1 'S)-2-(1-Acetamido-2-acetamido)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Hydrochloride The title compound was prepared according to the method described in Example 1 K, substituting (±)-(2R,3S,5R, 1'S)-2-(1,2-di-acetamido)butyl-3-vinyl pyrrolidine-5-carboxylic acid Hydrochloridesalt in place of (±)(2R,3R,5R, I1'S)-2-(1 acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 3.30 mg, 80%). MS: (M+H) =284, (M-H)=282, (M+CI)=318 Example 143 (±)-(2R,3S,5R,1'S)-2-(1-Acetamido-2-azido)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloride = AcHN, OtBu N
N
3 143A (±)-(2R,3S,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-N-acetamido-2-azido)ethyl-3 vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1 'S)-1 -t-Butoxycarbonyl-2-(N-acetylaziridinyl)-3-vinyl-pyrrolidine-5 carboxylic acid t-butyl ester (21.6 mg, 0.064 mmole) was reacted with sodium azide (41.6 mg, 0.64 mmole) and ammoniun chloride (34.2 mg, 0.64 mmol) in -357- WO 99/54299 PCT/US99/07945 ethanol (270 gL) and water (30 gL) at 750C for 1 hour. The ethanol was then removed in vacuo and the remaining aqueous was extracted with ethyl acetate. The combined organics were washed with brine, dried over MgSO 4 , and concentrated in vacuo (crude yield: 20mg, 82%). To the crude mixture was added acetic anhydride (31gL, 0.33 mmol) and triethylamine (92 gL, 0.66 mmol) in dichloromethane (330 gL) at 00C for 30 minutes. The reaction mixture was then concentrated in vacuo. The residue was purified by chromatography on silica gel using 100% dichloromethane to 50% dichloromethane/ethyl acetate to provide the title compound (yield: 10 mg, 60%). 1 H NMR (DMSO-d 6 )(rotamers) 6 7.85 and 7.81(d, J=9.5Hz and 9.8Hz, 1H), 5.94-5.80(m, 1H), 5.04-4.93(m, 2H), 4.58-4.38(m, 1H), 4.04-3.96(m, 1H), 3.72-3.66(m, 1H), 3.41-3.21(m, 2H), 3.09-2.79(m, 1H), 2.59-2.46(m, 1H), 1.84 1.82(m, 3H), 1.79-1.53(m, 1H), 1.43-1.35(m, 18H). MS: (M+H) =424, (M+Na) =446, (2M+Na)+=869, (M-H)-=422, (M+CI)=458 = AcHN. OH HH
N
3 HCI 143B (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-azido)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Hydrochloric Salt. The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(l1-acetamido 2-azido)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)(2R,3R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 2.94 mg, 93%). -358- WO 99/54299 PCT/US99/07945 1 H NMR (DMSO-d 6 ) 5 8.24(d, J=8.55Hz, 1H), 5.74-5.67(m, 1H), 5.14(d, J=17.1Hz, 1H), 5.06(d, J=10.4Hz, 1H), 4.41-4.35(m, 2H), 3.57-3.36(m, 3H), 2.93 2.90(m, 1H), 2.44-2.38(m, 1H), 1.96-1.84(m, 1H), 1.84(s, 3H). MS: (M+H)*=268, (M-H)-=266, (M+CI)=302 Example 144 (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-N-methylamino)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Dihydrochloride AcHN, O t Bu HN
CH
3 144A (±)-(2R.,3S.5R,1'S)-1l-t-Butoxvcarbonyl-2-(1-N-acetamido-2-N methylamino)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. Methylamine (.016 g, .53 mmole) was reacted with N,O-bis trimethylsilylacetamide (.079 g, .39 mmole) in DMSO (0.8 mL) at 00C for 1 hour. (±)-(2R,3S,5R, 1'S)-1 -t-Butoxycarbonyl-2-(N-acetylaziridinyl)-3-vinyl-pyrrolidine-5 carboxylic acid t-butyl ester (.040 g, .11 mmole) was then reacted with the above reagent N-trimethylsilylmethylamine at 750C for 18 hours. The reaction was diluted with ethyl acetate (7 mL) washed with water and brine, dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using chloroform-methanol-ammonia to provide the title compound (yield: .011 g, 25%). -359- WO 99/54299 PCT/US99/07945 1 H NMR (CDCI 3 ) 5 5.78-5.98 (m,1H), 5.90-5.04 (2m, 2H), 4.40-4.55 (brm, 1H), 3.90-4.02 (m, 1H), 3.64-3.75 (2m, 1H), 2.25-2.40 (brm 3H), 2.83,2.85 (2d, 3H), 1.42,1.44 (2s, 9H), 1.34,1.37 (2s, 9H). MS: (M+H)*= 412 AcHN ),OH HN 0 6 2 HCI
CH
3 144B (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-N-methylamino)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Dihydrochloride Salt The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1-acetamido 2-N-methylamino)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)(2R,3R,5R, 1'S)-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl-pyrrolidine 5-carboxylic acid t-butyl ester (yield: 7.2 mg, 99%). 1 H NMR (DMSO-d 6 ) 8 8.25 (d, 1H), 5.70 (m, 1H), 5.10 (m, 2H), 4.50 (m,1H), 4.40 (m,1H), 2.55 (s, 3H), 1.85 (s, 3H). MS: (M+H)*= 256 -360- WO 99/54299 PCT/US99/07945 Examples 145-164 AN OtBu 2. 6NHCI AcHN. OH AcN BOc 0 R'RN 0 2 HCI The following title compounds were prepared according to the methods described in Examples 141-144 where R' is equal to hydrogen. Where R or R' are not equal to hydrogen the corresponding amine is used directly without the intermediacy of trimethylsilylation. Example 145 AcHN OH ' N N H0 H- 2HCI (+)-(2R,3S,5R,1'S)-2-(1-Acetamido-2-N-isopropylamino)ethyl-3-vinyl-pyrrolidine 5-carboxylic Acid Dihydrochloride Salt 1 H NMR (DMSO-d 6 ) 5 8.30 (d, 1H), 5.70 (m, 1H), 5.10 (m, 2H), 4.40 (br, 2H), 3.52-3.68 (br, 1H), 3.10-3.20 (br, 1H), 2.82-2.97 (br, 1H), 2.37-2.47 (br, 1H), 1.88 (s, 3H), 1.25 (d, 6H). MS: (M+H) = 284 -361- WO 99/54299 PCT/US99/07945 Example 146 AcHN, OH 'N H N H 0 2HCI (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-N-butylamino)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Dihydrochloride Salt 1 H NMR (DMSO-d 6 ) 6 8.25 (d, 1H), 5.70 (m, 1H), 5.10 (m, 2H),4.50 (m, 1H), 4.38 (m, 1H), 3.60 (m, 1H), 2.90 (m, 3H), 2.40 (m, 2H), 1.87 (s, 3H), 1.62 (m, 2H), 1.33 (m, 2H), 0.90 (t, 3H). MS: (M+H) = 298 Example 147 AcHN ,OH ' N jj HN Ph 2HCI (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-N-benzylamino)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid HydrochlorideSalt 1 H NMR (DMSO-d 6 ) 6 7.56-7.43(m, 5H), 5.74-5.67(m, 1H), 5.15-4.99(m, 2H), 4.56(m, 1H), 4.27-3.93(m, 3H), 3.66-3.15(m, 3H), 2.91-2.88(m, 1H), 2.64 2.34(m, 2H), 1.86(s, 3H). MS: (M+H) =332, (M+Na) =354, (M-H)-=330, (2M-H)=661 -362- WO 99/54299 PCT/US99/07945 Example 148 AcHN. 7P KO HN O 0 HN Ph-_ 2HCI (±)-(2R,3S,5R. 1 'S)-2-(1-Acetamido-2-N-phenethylamino)ethyl-3-vinyl-pyrrolidine 5-carboxylic Acid Dihydrochloride Salt 1 H NMR (DMSO-d 6 ) 6 8.25 (d, 1H), 7.30 (m, 5H), 5.70 (m, 1H), 5.10 (m, 2H), 4.50 (br, 1H), 4.35 (br, 1H), 3.61 (m, 1H), 3.17 (m, 3H), 2.98 (m, 3H), 2.42 (m, 1H), 1.88 (s, 3H). MS: (M+H) = 346 Example 149 AcHN, OH -N 0 I 2HCI (±)-(2R,3S,5R.1 'S)-2-(1-Acetamido-2-N,N-dimethylamino)ethyl-3-vinyl-pyrrolidine 5-carboxylic Acid Dihydrochloride Salt 1 H NMR (DMSO-d 6 ) 6 8.34(d, J=9.2Hz, 1H), 5.74-5.67(m, 1H), 5.12(d, J=17.1Hz, 1H), 5.04(d, J=10.4Hz, 1H), 4.67-4.62(m, 1H), 4.40(dd, J=7.3, 10.4Hz, 1H), 3.60-3.11(m, 3H), 2.96-2.83(m, 1H), 2.50(s, 6H), 2.44-2.38(m, 1H), 1.92 1.84(m, 1H), 1.84(s, 3H). -363- WO 99/54299 PCT/US99/07945 MS: (M+H) =270, (M+Na)*=292, (M-H)=268. Example 150 AcHN. OH N H 0 2HCI (±)-(2R,3S, 5R,1'S)-2-(1 -Acetamido-2-N,N-diethylamino)ethyl-3-vinvI-pyrrolidine 5-carboxylic Acid Dihydrochloride Salt 1 H NMR (DMSO-d 6 ) 8 8.23 (d, 1H), 5.70 (m, 1H), 5.10 (m, 2H), 4.60 (br, 1H), 4.40 (br, 1H), 3.12 (m, 4H), 2.88 (m, 1H), 2.42 (m, 1H), 1.85 (s, 3H), 1.22 (t, 3H). MS: (M+H) = 298 -364- WO 99/54299 PCT/US99/07945 Example 151 AcHN OH H "/-N 2HCI (±)-(2R,3S,5R, 1 'S)-2-( 1 -Acetamido-2-N,N-dibutylamino)ethyl-3-vinyl-pyrrolidine 5-carboxylic Acid Dihydrochloride Salt 1 H NMR (DMSO-d 6 ) 5 8.24 (d, 1H), 5.70 (m, 1H), 5.08 (m, 2H), 4.48-4.62 (br, 1H), 4.28-4.43 (1H), 3.05 (m, 4H), 2.77-2.92 (br, 1H), 2.34-2.46 (br, 2H), 1.84 (s, 3H), 1.64 (m, 4H), 1.30 (m, 4H), 0.93 (t, 6H). MS: (M+H)*= 354 Example 152 = AcHN. OH N HN OH 2HCI (±)-(2R,3S,5R,1'S)-2-(1-Acetamido-2-(N-2-hydroxyethylamino))ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Dihydrochloride Salt 1 H NMR (DMSO-d 6 ) 8 8.20 (d, 1H), 5.70 (m, 1H), 5.15 (d, 1H), 5.08 (d, 1H), 4.50 (brm, 1H), 4.38 (brm, 1H), 3.68 (M, 1H), 3.0 (brm, 2H), 2.90 (m, 1H), 2.41 (m, 1H), 1.85 (s, 3H). -365- WO 99/54299 PCT/US99/07945 MS: (M+H) = 286 Example 153 AcHN. OH N-- OH 2HCI (±)-(2R,3S,5R. 1 'S)-2-(1 -Acetamido-2-(N-2-hydroxyethyl-N-ethylamino))ethyl-3 vinyl-pyrrolidine-5-carboxvlic Acid Dihydrochloride Salt 'H NMR (DMSO-d 6 ) 8 5.81-5.74(m, 1H), 5.38(d, J=17.1Hz, 1H), 5.22(d, J=10.0Hz, 1H), 4.92-4.88(m, 1H), 4.48(dd, J=7.6, 9.8Hz, 1H), 3.91(t, J=4.9Hz, 2H), 3.85(dd, J=5.6, 10.0Hz, 1H), 3.63-3.53(m, 2H), 3.46-3.39(m, 4H), 3.16 3.13(m, 1H), 2.66-2.61(m, 1H), 2.08(s, 3H), 2.06-2.01(m, 1H), 1.38(t, J=7.33, 3H). MS: (M+H)*=314, (M+Na)+=336, (M-H)-=312, (M+CI)-=348, (2M-H)-=625 -366- WO 99/54299 PCTIUS99/07945 Example 154 AcHN, OH 0N OH 2HCI (±)-(2R, 3S, SR,1I'S)-2-(l1-Acetamido-2-(N-2-hvdroxyethyl-N-Dropylamino))ethyl-3 vinvl-Dyrrolidine-5-carboxylic Acid Dihyd rochioride Salt 1 H NMR (DMSO-d 6 ) 6 8.36(d, J=8.5Hz, 1IH), 5.75-5.68(m, 1IH), 5.13(d, J=1 7.1lHz, 1 H), 5.04(d, J= O.4Hz, 1 H), 4.62(m, 1 H), 4.36(m, 1 H), 3.77(t, J=4.9Hz, 2H), 3.63-3.59(m, 1IH), 3.50-3.23(m, 3H), 3.22-3.19(m, 2H), 3.08(t, J=7.3Hz, 2H), 2.91-2.87(m, 1IH), 2.44-2.39(m, 1 H), 1.99-1.88(m, 1IH), 1.84(s, 3H), 1.75-1.70(m, 2H), 0.90(t, J=6.7Hz, 3H). MS: (M+H)'=328, (M+Na)+=35O, (M-H)=326, (M+Cl)=362, (2M-H)=653 Example 155 AcHN. * H N , 2HCI (±)-(2R. 3S,5R, 1'S)-2-(l1-Acetamido-2-(imidazol-1 -vi))ethyl-3-vinvl-pyrrolidine-5 carboxylic Acid DiHydrochloride 1 H NMR (MeOD-d 3 ) 6.9.06(s, 1 H), 7.72(s, 1 H), 7.58(s, 1 H), 5.84-5.76(m, 1 H), 5.39(d, J=1 7.1lHz, 1 H), 5.23(d, J=1O0.25Hz, 1 H), 4.70-4.66(m, 1 H), 4.52 -367- WO 99/54299 PCT/US99/07945 4.43(m, 2H), 3.92-3.89(m, 1H), 3.20-3.17(m, 1H), 2.67-2.62(m, 1H), 2.11-2.04(m, 1H), 1.95-1.89(m, 1H), 1.91(s, 3H). MS: (M+H) =293, (M-H)-=291, (M+35) =327. Example 156 AcHN' OH 0 N HO N .OH 2HCI HO OH (±)-(2R,3S,5R,1'S)-2-(l1-Acetamido-2-(N,N-di-(2-hydroxyethylamino))ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Dihydrochloride Salt MS: (M+H)+=330, (M+Na)+=352, (M-H)-=328, (M+CI)-=364 Example 157 = AcHN OH N AcN 0 S HCI (±)-(2R,3S,5R. 1'S)-2-(1 -Acetamido-2-(N-acetyl-N-methylamino)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (DMSO-d 6 ) 8 8.01,7.95 (2d, 1H), 5.68-5.80 (m,1H), 5.02-5.22 (m, 2H), 4.30-4.45 (brm, 2H), 3.26,3.21 (2d, 1H), 2.82-2.95 (brm, 1H), 2.38-2.48 (m, 1H), 1.98,2.02 (2s, 3H),1.79,1.82 (2s, 3H). MS: (M+H)
+
= 298 -368- WO 99/54299 PCT/US99/07945 Example 158 AcHN, OH
CH
3 N10 K,,/OH 2HCI (±)-(2R,3S, 5R, 1 'S)-2-( 1 -Acetamido-2-(N-2-hydroxyethl-N-methylamino))ethl-3 vinyl-pyrrolid ine-5-carboxylic Acid Dihyd rochioride Salt 1 H NMR (DMSO-d 6 ) 8 8.35(d, J=9.l5Hz, 1 H), 5.74-5.67(m, 1IH), 5.12(d, J=1 7.1lHz, 1IH), 5.04(d, J=1 .4Hz, 1 H), 4.70(m, 1IH), 4.39(dd, J=7.3, 10.4Hz, 1 H), 3.80-3.75(m, 3H), 3.61-3.43(m, 3H), 3.23-3.16(m, 2H), 2.91-2.82(m, 1 H), 2.82(s, 3H), 2.44-2.39(m, 1H), 1.92-1.84(m, 1H), 1.84(s, 3H). MS: (M+H)+=300, (M+Na)+=322, (2M+H-H 2 )=58 1 Example 159 AcHN, ,OH ' N -K
CH
3 N 0 KI/ 2HCI (±)-(2R,3S, 5R,1I'S)-2-(1 -Acetamido-2-(N-ropvrl-N-methlamino)ethyl-3-vinvl pyrrolidine-5-carboxylic Acid Dihydrochioride Salt 1 H NMR (DMSO-d 6 ) (broad) 8 8.3(l1H), 5.7(l1H), 5.12-5.04(2H), 4.6(l1H), 4.35(1 H), 2.61-2.35(1 1 H), 1 .9(3H), 1.78-1 .63(2H), 1 .9(3H). MS: (M+H)+=298, (M+Na)+=320, (M-H)=296, (M+Cl)332, (2M-H)593 -369- WO 99/54299 PCT[US99/07945 Example 160 AcH N, OH
CH
3 N0 6 2HCI (±)-(2R,3S .5R1 l'S)-2-( 1 -Acetamido-2-(N-cyclohexyl-N-methylamino))ethvl-3-vinyl pvrrolid ine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (DMSO-d 6 ) 5 8.26(m, 1 H), 5.75-5.65(m, 1IH), 5.08(d, J=1 7.1lHz, 1 H), 5.02(d, J= 10. 3Hz, 1IH), 4.62(m, 1 H), 4.43-4.40(m, 1 H), 3.62-3.58(m, 1IH), 3.46-3.16(m, 2H), 2.89-2.84(m, 1IH), 2.72(s, 3H), 2.44-2.39(m, 1 H), 2.07-1.80(m, 5H), 1.81 (s, 3H), 1.63(m, 1 H), 1.45-1.06(m, 6H). MS: (M+H)+=338, (M+Na)+=360, (M-H)-=336, (M+CI)-=372 Example 161 AcHN, OH CH3N 2HCI0 Ph (±)-(2R. 3S,5R, 1'S)-2-(1 -Acetamido-2-(N-benzyl-N-methylamino))ethl-3-vinvl Pyrrolidine-5-carboLxvlic Acid Dihydrochloride Salt 1 H NMR (DMSO-d 6 ) 6 8.36(m, 1H), 7.61-7.46(m, 5H), 5.69-5.64(m, 1H), -370- WO 99/54299 PCT/US99/07945 4.25(d, J=12.9, 1H), 3.61(m, 1H), 3.43(m, 1H), 3.22(m, 1H), 2.93-2.85(m, 1H), 2.73(s, 3H), 2.44-2.38(m, 1H), 1.92-1.85(m, 1H), 1.85(s, 3H). MS: (M+H) =346 Example 162 = AcHN. OH ' N
CH
3 N P ph 2HCI (+)-(2R,3S,5R. 1 'S)-2-(1-Acetamido-2-(N-phenethyl-N-methylamino))ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Dihydrochloride Salt 1 H NMR (DMSO-d 6 ) 8 8.34(d, J=8.55Hz, 1H), 7.37-7.26(m, 5H), 5.76 5.69(m, 1H), 5.14(d, J=17.1Hz, 1H), 5.06(d, J=10.4Hz, 1H), 4.72(m, 1H), 4.46 4.42(m, 1H), 3.83-3.20(m, 6H), 3.13-2.99(m, 2H), 2.86(s, 3H), 2.95-2.83(m, 1H), 2.46-2.40(m, 1H), 1.95-1.81(m, 1H), 1.86(s, 3H). MS: (M+H) =360 -371- WO 99/54299 PCT/US99/07945 Example 163 AcHN. OH ' N
CH
3 N 02HCI (±)-(2 R. 3S, 5 R, I 'S)-2-(1 -Acetam id o-2-(N -nap hthyl methvl-N -methyl am ino))ethl-3 vinvl-pyrroldine-5-carboxylic Acid Dihyd rochioride Salt 1 H NMR (DMSO-d 6 ) 5 8.41 (d, J=7.3Hz, 1 H), 8.32-7.59(m, 7H), 5.60(m, 1 H), 5.04(d, J=1 7.1lHz, 1 H), 4.91 (d, J=9.8Hz, 1 H), 4.97-4.73(m, 3H), 4.39(m, 1IH), 3.70-3.13(m, 3H), 2.90(m, 1 H), 2.72(s, 3H), 2.43-2.41 (m, 1 H), 2.01-1.74(m, 1 H), 1 .87(s, 3H). MS: (M+H)+=395, (M+Na)+=41 8, (M-H)=394, (M+CI)-=43O, (2M-H)=789 -372- WO 99/54299 PCT/US99/07945 Example 164 AcHN ,OH N O 2HCI (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-(N-morpholinyl))ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Dihydrochloride Salt 1 H NMR (DMSO-d 6 ) 8 8.28 (d, 1H), 5.75-5.78 (m, 1H), 5.15 (d,1H), 5.05 (d, 1H), 4.65 (brm, 1H), 4.42 (m, 1H), 3.72-3.98 (brm, 3H), 3.62 (m, 1H), 2.90 (m, 1H), 2.38-2.48 (m, 1H), 1.85 (s, 3H). MS: (M+H) = 312 -373- WO 99/54299 PCT/US99/07945 Example 165 (±)-(2R,3S,5R, 1 'S,3'R)-2-(1 -Acetamido-2-(N-methyl-N-t-butylamino-N oxide))ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloride Salt = = AcHN OtBu AcHN, OtBu N I N N H Boc N H Ko 4 CH 3 O 'CH 3 165A (±)-(2R.3S,5R,1'S,3'R) and (±)-(2R,3S,5R,1'S,3'S)-1l-t-Butoxycarbonyl-2 (1-acetamido-2-(N-methyl-N-t-butylamino-N-oxide))ethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1'S)-1 -t-Butoxycarbonyl-2-(1 -acetamido-2-(N-methyl-N-t butylamino))ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (37 mg, .08 mmole) was reacted with the m-chloroperoxybenzoic acid (20 mg, .08 mmole) in
CH
2
CI
2 (0.9 mL) at 0OC for 1 hour. The reaction was chromatographed directly on silica gel eluting with a gradient of acetone to acetone/30% MeOH to provide the title compounds isomer (±)-(2R,3S,5R,1'S,3'R) (yield: .010 g, 27%) and isomer (±)-(2R,3S,5R,1'S,3'S) (yield: .011 g, 29%). = AcHN. N OH N H 0 / CH 3 HCI O 0 165B (±)-(2R.3S,5R,1'S,3'R)-2-(1-Acetamido-2-(N-methyl-N-t-butylamino-N oxide))ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloride Salt The title compound was prepared according to the method described in Example 15C substituting (±)-(2R,3S,5R, 1 'S,3'R)-1-t-butoxycarbonyl-2-(1 -374- WO 99/54299 PCT/US99/07945 acetamido-2-(N-methyl-N-t-butylamino-N-oxide))ethyl-pyrrolidine-5-carboxylic acid t-butyl ester for (±)-(2R,3S,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3 (imidazol-2-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 6 mg, 80%). 1 H NMR (CD 3 OD) 5 5.72-5.87 (m, 1H), 5.40 (d, 1H), 5.20-5.28 (m, 2H), 4.44-4.53 (dd, 1H), 3.73-3.95 (m, 3H), 3.57 (s, 3H), 3.08-3.19 (m, 1H), 2.59-2.72 (m, 1H), 2.05-2.15 (m, 1H), 2.04 (s, 3H), 1.54 (s, 9H). MS: (M+H) = 328 Examples 166-178 -'1. MCPBA 2. chromatographic AcHN. OH separation AcHN. OH C P ' 3. 6NHCI O H N R'RN O R'RN HCI The following title compounds were prepared according to the method described in Example 165. -375- WO 99/54299 PCT/US99/07945 Example 166 = AcHN. OH 0'N HH N HCI O 'CH 3 (±)-(2R,3S,5R,1'S.,3'R)-2-(1-Acetamido-2-(N-methyl-N-isopropylamino-N oxide))ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (MeOD-d3) 8 5.87-5.74 (m, 1H), 5.46-5.40 (m, 1H), 5.27-5.23 (m, 1H), 5.21-5.18 (m, 1H), 4.50 (dd, J=8.1, 9.8Hz, 1H), 4.04-3.87 (m, 4H), 3.54 (s, 3H), 3.20-3.14 (m, 1H), 2.69-2.60 (m, 1H), 2.12-2.01 (m, 1H), 2.05 (s, 3H), 1.50 (d, J=6.4Hz, 3H), 1.48(d, J=6.4Hz, 3H). MS: (M+H)+=314, (M+Na)+=336, (2M+1)
+
= 627, (2M+Na)+=649. Example 167 = AcHN OH 0NN HH " d -'CH 3 HC (±)-(2R,3S,5R,1'S,3'S)-2-(1-Acetamido-2-(N-methyl-N-propylamino-N-oxideethyl 3-vinyl-pyrrolidine-5-carboxylic Acid HydrochlorideSalt 1 H NMR (MeOD-d3) 8 5.87-5.74 (m, 1H), 5.46-5.40 (m, 1H), 5.27-5.23 (m, 1H), 5.21-5.18 (m, 1H), 4.50 (dd, J=8.1, 9.8Hz, 1H), 4.04-3.87 (m, 4H), 3.54 (s, -376- WO 99/54299 PCT/US99/07945 3H), 3.20-3.14 (m, 1H), 2.69-2.60 (m, 1H), 2.12-2.01 (m, 1H), 2.05 (s, 3H), 1.50 (d, J=6.4Hz, 3H), 1.48(d, J=6.4Hz, 3H). MS: (M+H) =314, (M+H-H 2 0)-=295 Example 168 AcHN- P ,OH N I HHO 4 CH 3 (+)-(2R,3S,5R, 1 'S,3'S)-2-(1 -Acetamido-2-(N-methyl-N-ethylamino-N-oxide))ethyl 3-vinyl-pyrrolidine-5-carboxylic Acid HydrochlorideSalt 1 H NMR (MeOD-d 3 ) 8 5.82-5.75 (m, 1H), 5.44(d, J=17.1Hz, 1H), 5.26(d, J=10.4Hz, 1H), 5.14-5.11(m, 1H), 4.48-4.45(m, 1H), 4.9(d, J=4.9Hz, 2H), 3.87(dd, J=4.9, 10.4Hz, 1H), 3.76(q, J=6.7Hz, 2H), 3.54(s, 3H), 3.17-3.09(m, 1H), 2.68-2.62 (m, 1H), 2.06(s, 3H), 2.09-2.03 (m, 1H), 1.45(t, J=7.3Hz, 3H). MS: (M+H)*=300, (M+Na) =322, (M+H-H 2 0)'=282 -377- WO 99/54299 PCT/US99/07945 Example 169 AcHN. OH N 0 HH o N: d CH 3 HCI (±)-(2R,3S,5R. 1 'S)-2-(1-Acetamido-2-(N,N-dimethylamino-N-oxide)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (DMSO-d 6 ) 6 8.58 (d, 1H), 5.67-5.78 (m, 1H), 5.20 (d, 1H), 5.08 (d, 1H), 4.62-4.78 (brm, 1H), 4.25-4.42 (brm, 1H), 4.06 (d, 1H), 3.85-3.95 (brm, 1H), 3.88-3.98 (brm, 1H), 3.35-3.50 (brs, 6H), 2.36-2.48 (m, 1H), 1.92 (m, 1H), 1.85 (s, 3H). MS: (M+H) = 286 Example 170 AcHN, OH HH O Ph N HCI O "CH3 (±)-(2R,3S,5R,1'S,3'S)-2-(1-Acetamido-2-(N-methyl-N-benzylamino-N oxide))ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid HydrochlorideSalt 1 H NMR (MeOD-d 3 ) 5 7.60-7.47(m, 5H), 5.75-5.65(m, 1H), 5.39(d, J=6.35Hz, 1H), 5.21(d, J=8.8Hz, 1H), 5.18-5.11(m, 1H), 5.00-4.70(m, 2H), 4.35 -378- WO 99/54299 PCT/US99/07945 4.27(m, 1H), 4.00-3.94(m, 2H), 3.86-3.79(m, 1H), 3.20(s, 3H), 3.14-3.05(m, 1H), 2.77-2.50(m, 1H), 2.08(s, 3H), 2.10-2.94(m, 1H). MS: (M+H)+=362, (M+Na)+=385, (M-H)-=360, (M+35)-=396 Example 171 = AcHN OH HN O' OH0
ONCH
3 HCI (±)-(2R,3S,5R, 1 'S,3'S)-2-(1 -Acetamido-2-(N-methyl-N-t-butylamino-N-oxide)ethyl 3-vinyl-pyrrolidine-5-carboxylic Acid HydrochlorideSalt 1 H NMR (CD 3 OD) 6 5.80 (m, 1H), 5.44 (d, 1H), 5.27 (d, 1H), 5.08 (m, 1H), 4.34-4.44 (dd, 1H), 3.83-3.94 (m, 3H), 3.38 (s, 3H), 3.02-3.18 (m, 1H), 2.58-2.72 (m, 1H), 2.08 (s, 3H), 1.97-2.08 (m, 1H), 1.55 (s, 9H). MS: (M+H)*= 328 -379- WO 99/54299 PCT/US99/07945 Example 172 = AcHN.- OH N N O(
H
0 N O" CH 3 HCI (±)-(2R,3S,5R,1'S,3'S)-2-(1 -Acetamido-2-(N-methyl-N-isopropylamino-N oxide))ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (MeOD-d 3 ) 6 5.86-5.74 (m, 1H), 5.53-5.47 (m, 1H), 5.29-5.25 (m, 1H), 5.22-5.19 (m, 1H), 4.50 (dd, J=8.1, 9.5Hz, 1H), 4.13-4.04 (m, 2H), 3.96 (dd, J=4.1, 10.5Hz, 1H), 3.87-3.82 (m, 1H), 3.39 (s, 3H), 3.23-3.17 (m, 1H), 2.70-2.61 (m, 1H), 2.11 (s, 3H), 2.08-2.00 (m, 1H), 1.50 (d, J=6.4Hz, 3H), 1.49(d, J=6.4Hz, 3H). MS: (M+H)+=314, (M+Na)+=336, (2M+1)+=627, (2M+Na)+=649. Example 173 AcHN ,OH
H
0 o'CH 3 HO (±)-(2R,3S,5R,1'S,3'R)-2-(1-Acetamido-2-(N-methyl-N-propylamino-N oxide))ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloride Salt 'H NMR (MeOD-d3) 6 5.82-5.75(m, 1H), 5.45(d, J=17.1Hz, 1H), 5.26(d, J=10.4Hz, 1H), 5.07-5.13(m, 1H), 4.48-4.42(m, 1H), 3.98(d, J=5.5Hz, 2H), -380- WO 99/54299 PCT/US99/07945 3.86(dd, J=4.3, 9.8Hz, 1H), 3.67-3.64(m, 2H), 3.46(s, 3H), 3.16-3.01(m, 1H), 2.68-2.62(m, 1H), 2.09-2.02(m, 1H), 2.06(s, 3H), 1.92-1.86(m, 2H), 1.04(t, J=7.3Hz, 3H). MS: (M+H)*=314, (M+H-H 2 0)=295 Example 174 = AcHN. H N O H N & 'CH 3 HCI (+)-(2R,3S,5R, 1 'S,3'R)-2-(1 -Acetamido-2-(N-methyl-N-ethylamino-N-oxide))ethyl 3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (MeOD-d 3 ) 8 5.82-5.75(m, 1H), 5.45(d, J=17.1Hz, 1H), 5.26(d, J=10.4Hz, 1H), 5.13-5.10(m, 1H), 4.48-4.44(m, 1H), 4.02-3.94(m, 2H), 3.89 (dd, J=4.3, 9.8Hz, 1H), 3.82(q, J=7.3Hz, 2H), 3.46(s, 3H), 3.18-3.10(m, 1H), 2.68-2.62 (m, 1H), 2.09(s, 3H), 2.07-2.02(m, 1H), 1.46(t, J=7.3Hz, 3H). MS: (M+H) =300, (M+Na)+=322, (M+H-H 2 0)*=282 -381- WO 99/54299 PCTIUS99/07945 Example 175 AcHN. P N OH H 0 Ph N "CHCI (±)-(2R.3S,5R, I'S, 3'S)-2-(1 -Acetamido-2-(N-methyl-N-benzyamino-N oxide))ethy1-3-vinyl-pyrrolidine-5-carboxylic Acid HydrochlorideSalt 'H NMR (MeOD-d 3 ) 6 7.60-7.47(m, 5H), 5.75-5.65(m, 1H), 5.39(d, J=6.35Hz, 1 H), 5.21 (d, J=8.8Hz, 1 H), 5.18-5.11 (m, 1 H), 5.00-470(m, 2H), 4.35 4.27(m, 1IH), 4.00-3.94(m, 2H), 3.86-3.79(m, 1 H), 3.40(s, 3H), 3.14-3.05(m, 1 H), 2.77-2.50(m, 1 H), 2.08(s, 3H), 2.10-2.94(m, 1IH). MS: (M+H)'=362, (M+Na)+=385, (M-H)-=360, (M+35)-=396 Example 176 AcHN. P N OH H 0 HCI 0 (±)-(2R,35. 5R, 1'S)-2-(1 -Acetamido-2-(N N-diethylamino-N-oxide))ethl-3-vinyl rgyrrolidine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (MeOD-d 3 ) 6 5.84-5.78(m, 1 H), 5.45(d, J1l6.85Hz, 1IH), 5.26(d, J=1 O.OHz, 1 H), 5.09-5.05(m, 1 H), 4.45-4.42(m, 1 H), 3.96-3.86(m, 3H), 3.76(q, -382- WO 99/54299 PCT/US99/07945 J=6.6Hz, 2H), 3.70(q, J=7.3Hz, 2H), 3.15-3.11(m, 1H), 2.68-2.62(m, 1H), 2.08 2.02(m, 1H), 2.08(s, 3H), 1.44-1.38(m, 6H). MS: (M+H)+=314, (M+Na) =336, (M+2Na)+=358 Example 177 = AcHN OH ~ 'N N HH O II !11 0 (±)-(2R,3S, 5R, 1 'S,3'R)-2-(1-Acetamido-2-(N-pyrrolidinyl-N-oxide))ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (DMSO-ds) 5 8.74 (d, 1H), 5.65-5.80 (m,1H), 5.28 (d, 1H), 5.10 (d,1H), 4.82 (m, 1H), 4.40-4.50 (dd, 1H), 4.30 (d, 1H), 3.60-4.12 (brm, 5H), 2.98 3.15 (m, 1H), 2.38-2.48 (m, 1H), 2.05-2.20 (brm, 5H), 1.88-1.98 (m, 1H), 1.87 (s, 3H). MS: (M+H) = 312 -383- WO 99/54299 PCT/US99/07945 Example 178 = AcHN. OH N PH0 N"O HCI 0,, (+)-(2R,3S,5R,1'S)-2-(1-Acetamido-2-(N-morpholinvyl-N-oxide)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (DMSO-d 6 ) 8 8.65 (d, 1H), 5.66-5.80 (m, 1H), 5.22 (d, 1H), 5.09 (d, 1H), 4.78 (brs, 1H), 4.32-4.42 (dd, 1H), 4.10-4.17 (brm, 2H), 3.50-4.02 (brm, 9H), 2.92-3.04 (brm, 1H), 2.37-2.48 (m, 1H), 1.88-1.96 (m, 1H), 1.87 (s, 3H). MS: (M+H) = 328 Example 179 (±)-(2R,3S,5R,1'S,3'S)-2-(1-Acetamido-2-(N-ethyl-N-methylamino-N-oxide))ethyl 3-(cis-Dropen-1 -yl)-pyrrolidine-5-carboxylic Acid HydrochlorideSalt O'Bu B 0 179A (±)-(2R,3S,5R, l'S)-1-t-Butoxycarbonyl-2-oxiranyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 1231, substituting ethyltriphenylphosphonium bromide in place of methyltriphenylphosphonium bromide (yield: 350 mg, 77%). -384- WO 99/54299 PCT/US99/07945 1 H NMR (CDCl 3 ) (rotamers) 6 5.55-5.43 (m, 2H), 4.13-4.04 (m, 2H), 3.14 3.11 (m, 2H), 2.76-2.50 (m, 3H), 1.75-1.70 (m, 1H), 1.64(d, 3H), 1.48-1.43(m, 18H). MS: (M+H)+=354, (M+Na) +=376, (2M+Na) =729 MsO H'Bu~ N
N
3 O 179B (±)-(2R.3S,5R, 1 'R)-l-t-Butoxvcarbonyl-2-(1-methanesulfonyloxy-3 azido)ethyl-3-(cis-propen-1 -ylv)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 123J, substituting (±)-(2R,3S,5R,1'S)-l1-t-butoxycarbonyl-2-oxiranyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3S,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido)butyl-3-vinyl-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 1.08 g, 84%). 1 H NMR (DMSO-d 6 ) (rotamers) 6 5.53-5.33 (m, 2H), 5.05-4.93 (m, 1H), 4.20-3.90 (m, 2H), 3.76-3.62 (m, 2H), 3.24 (s, 3H), 2.59-2.49(m, 1H), 1.64 1.55(m, 5H), 1.43-1.36(m, 18H). MS: (M+H) =475, (M+Na)+=497, (2M+Na)+=971 -385- WO 99/54299 PCT/US99/07945 H OtBu HN 'N iK BC0 179C (±)-(2R, 3S,5R, 1 'S)-1 -t-Butoxycarbonyl-2-aziridinyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 123K, substituting (2R,3S,5R, 1 'S)-1 -t-butoxycarbonyl-2-(1 methanesulfonyloxy-3-azido)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (2R,3S,5R, 1 'S)-1l-t-butoxycarbonyl-2-(1 methanesulfonyloxy-3-azido)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester(crude yield: 564 mg, 71%). 'H NMR(DMSO-d 6 ) (rotamers) 6 5.45-5.30 (m, 2H), 4.15-3.99 (m, 1H), 3.30-3.08 (m, 1H), 3.07-2.84 (m, 1H), 2.68-2.51 (m, 1H), 2.13-1.85(m, 1H), 1.80 1.05(m, 3H), 1.57(d, J=5.4Hz, 3H), 1.41-1.35(m, 18H). MS: (M+H)+=352, (M+23) =375, (2M+H) =705, (2M+23)+=727 AcN TP OtBu B 0 179D (±)-(2R.3S,5R, l1'S)-1 -t-Butoxycarbonyl-2-(N-acetylaziridinyl)-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 123L, substituting (±)-(2R,3S,5R, l'S)- 1 -t-butoxycarbonyl-2-aziridinyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) -386- WO 99/54299 PCT/US99/07945 (2R,3S,5R, 1'S)-1 -t-butoxycarbonyl-2-azirid inyl-3-vinyl-pyrrolid ine-5-carboxylic acid t-butyl ester(yield: 455 mg, 72%). 1 H NMR(DMSO-d 6 ) (rotamers) 6 5.74-5.34(m, 2H), 4.17(dd, J=2.4, 6.35Hz, 1H), 3.41(dd, J=1.95, 6.35Hz, 1H), 3.14-2.99(m, 1H), 2.73-2.58(m, 2H), 2.40(d, J=6.35Hz, 1H), 2.17-2.12(m, 1H), 2.05-2.00(m, 3H), 1.66-1.55(m, 1H), 1.56(d, J=6.8Hz, 3H), 1.41-1.31(m, 18H). MS: (M+H)+=395, (M+Na)+=417, (M+H+Na)+=418, /=, AcN.. OtBu N O N 179E (±)-(2R,3S,5R. 1'S)-l-t-Butoxycarbonyl-2-(1-acetamido-2-N-ethyl-N methylamino)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 150, substituting N-ethyl-N-methyl-amine in place of diethylamine (yield: 30 mg, 87%). MS: (M+H)+=454, (M+Na)+=476, (M-H)-=452, (M+35)-=488 -387- WO 99/54299 PCT/US99/07945 AcN. O'Bu AcN OtBu B N aoC' u BN" 0 B.~H 0 O CH 3 O CH 3 179E (±)-(2R,3S,5R,1'S,3'R) and (±)-(2R,3S,5R, 1 'S,3'S)-1l-t-Butoxycarbonyl-2 (1-acetamido-2-(N-ethyl-N-methylamino-N-oxide))ethyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 165A, substituting (±)-(2R,3S,5R,1'S)-l-t-butoxycarbonyl-2-(1 acetamido-2-(N-ethyl-N-methylamino))ethyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'S)-1-t-butoxycarbonyl-2 (1-acetamido-2-N-methyl-N-t-butylamino)ethyl-3-(cis-propen- 1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 15.2 mg, 51%).
H
3 C AcHN. OH 'N C
"
cH 3 179F (±)-(2R,3S,5R,1 'S,3'R)-2-(l1-Acetamido-2-(N-ethyl-N-methylamino-N oxide))ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Hydrochloride Salt The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3S,5R,1'S,3'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-(N-methyl-N-ethyl-N-oxide))ethyl-pyrrolidine-5-carboxylic acid t-butyl ester for (±)-(2R,3R,5R, l'S)-2-(1-acetamido-3-ethyl)pentyl-3-methoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester (yield: 8.7 mg, 29%). 1 H NMR (MeOD-d 3 ) 5 5.75-5.69(m, 1H), 5.37-5.30(m, 1H), 5.07-5.04(m, 1H), 4.49(dd, J=7.8, 10.2Hz, 1H), 4.05-3.74(m, 4H), 3.61-3.32 (m, 1H), 3.55(s, -388- WO 99/54299 PCT/US99/07945 3H), 2.69-2.60(m, 1H), 2.04(s, 3H), 1.95-1.84(m, 1H), 1.75(dd, J=2.0, 7.1Hz, 3H), 1.44 (t, J=7.1Hz, 3H). MS: (M+H)+=314, (M+35)+=348 Examples 179-184 1. MCPBA / 2. chromatographic AcHN. OH separation AcHN. OH N 3. 6 NHCI O H N R'RN O R'RN O HCI The following title compounds were prepared according to the method described in Example 179. Example 180
H
3 C AcHN. OH N H O N HCI O" "CH 3 (+)-(2R, 3S,5R, 1 'S,3'S)-2-(1 -Acetamido-2-(N-ethyl-N-methylamino-N-oxide))ethyl 3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (MeOD-d 3 ) 6 5.75-5.69(m, 1H), 5.38-5.30(m, 1H), 5.02-4.98(m, 1H), 4.47(dd, J=7.8, 9.8Hz, 1H), 4.02-3.77(m, 4H), 3.56-3.39(m, 1H), 3.47(s, 3H), 2.69-2.59(m, 1H), 2.07(s, 3H), 1.95-1.84(m, 1H), 1.76(dd, J=1.7, 7.1Hz, 3H), 1.46(t, J=7.1Hz, 3H). MS: (M+H)+=314, (M+35)*=348 -389- WO 99/54299 PCT/US99/07945 Example 181
H
3 C AcHN- *OH H0 N. HCI .~ CH 3 (±')-(2 R, 3S. 5R. 1VS. 3'R)-2-(1 -Acetam id o-2-(N-iso propyl-N -methyl a mino-N oxide))ethyl-3-(cis-P rope n- 1 -vl)-pyrro lid ine-5-carboxyl ic Acid Hyd rochloride Salt 1 H N MR (MeOD-d 3 ) 6 5.76-5.66(m, 1 H), 5.39-5.31 (m, 1IH), 5.17-5.11 (m, 1 H), 4.51 (dd, J=7.5, 10.2Hz, 1 H), 4.07-3.76(m, 4H), 3.55(S, 3H), 3.52-3.39(m, 1IH), 2.69-2.60(m, 1IH), 2.02 (S, 3H), 2.08-1.84(m, 1 H), 1.75(dd, J=1.7, 7.1 Hz, 3H), 1.50(d, J=6.lHz, 3H), 1.48(d, J=6.4Hz, 3H). MS: (M+H)'=314, (M+35)+=348 Example 182
H
3 C AcHN. ~ OH H0 N HCI 0"CH 3 (±)-(2 R,35 R, 1VS, 3'S)-2-(1 -Acetam id o-2-(N-isopropyl-N-methylam i n0- N oxide))ethyl-3-(cis-propen-1 -yI)-pyrrolidine-5-carboxylic Acid Hydrochloride Salt 'H NMR (MeQD-d 3 ) 5 5.76-5.68(m, 1H), 5.39-5.31(m, 1H), 5.10-5.05(m, 1 H), 4.49(dd, J=7.8, 9.8Hz, 1 H), 4.12-3.84(m, 4H), 3.55-3.44(m, 1 H), 3.41 (S, 3H), -390- WO 99/54299 PCT/US99/07945 2.69-2.60(m, 1IH), 2.08(S, 3H), 2.07-1.84(m, 1IH), 1.76(dd, J=1.7, 6.8Hz, 3H), 1.51(d, J=2.4Hz, 3H), 1.49(d, J=2.4Hz, 3H). MS: (M+H)=314, (M+35)+=348 Example 183
H
3 C AcHN. ~ H H 0 N "C3HCI (±)-(2R. 3S.5R, 1'S.3'S)-2-(lI-Acetamido-2-(N-isobutl-N-methylamilo-N oxide)ethyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboLxvlic Acid Hydrochloride Salt 1 H NMR (MeOD-d 3 ) 5 5.75-5.69(m, 1 H), 5.38-5.31 (m, 1 H), 5.18-5.12(m, 1IH), 4.53(dd, J=7.5, 9.8Hz, 1IH), 4.25-3.42(m, 6H), 3.65 (s, 3H), 2.68-2.58(m, 1H), 2.44-2.36(m, 1H), 2.05(s, 3H), 1.94-1.87(m, 1H), 1.76(d, J=2.7Hz, 3H), 1.14 (d, J=6.8Hz, 6H). MS: (M+H)=342, (M+Na)+=364, (M-H)-=340 -391- WO 99/54299 PCT/US99/07945 Example 184
H
3 C AcHN. OH CN CH 3 (+)-(2R,3S,5R, 1 'S,3'R)-2-(1 -Acetamido-2-(N-isobutyl-N-methylamino-N oxide))ethyl-3-(cis-propen-1 -yI)-pyrrolidine-5-carboxylic Acid Hydrochloride Salt 1 H NMR (MeOD-d 3 ) 6 5.75-5.69(m, 1H), 5.38-5.31(m, 1H), 5.06-5.02(m, 1H), 4.48(dd, J=7.5, 9.8Hz, 1H), 4.08-3.85(m, 3H), 3.70-3.57 (m, 2H), 3.52(s, 3H), 3.48-3.41(m, 1H), 2.70-2.60(m, 1H), 2.40-2.36(M, 1H), 2.08(s, 3H), 1.95 1.84(m, 1H), 1.75(dd, J=1.7, 7.1Hz, 3H), 1.14 (d, J=6.8Hz, 6H). MS: (M+H)+=342, (M+Na) =364, (M-H)-=340 -392- WO 99/54299 PCT/US99/07945 Example 185 (±)-(2R,3S, 5R, 1 'S)-2-(1 -Acetamido-2-(N-isopropyl-N-hydroxyamino))ethyl-3-vinyl pyrrolidine-5-carboxylic Acid HydrochlorideSalt = AcHN. OtBu NN o OH 165A (±)-(2R,3S,5R, I'S)-l-t-Butoxycarbonyl-2-(1-acetamido-2-(N-isopropyl-N hydroxvamino))ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R,1'S)-l-t-Butoxycarbonyl-2-(1-acetamido-2 isopropylamino)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (21 mg, 0.048 mmole) was dissolved in 0.95 mL of acetone. It was then titrated with 0.14 mL of a solution of dimethyldioxirane (0.1 M) in acetone at -45 0 C for 0.5 hour. The reaction was stopped by concentrating the mixture in vacuo. The residue was purified by chromatography on silica gel using 100% dichloromethane to 90% dichloromethane/methanol to provide the title compound (yield: 5.3 mg, 24%) and recovered starting material (yield 12 mg, 57%). 1 H NMR (MeOD-d3) 6 5.95-5.89(m, 1H), 5.08-4.94(m, 2H), 4.75-4.68(m, 1H), 4.13-3.83(m, 2H), 2.85-2.47(m, 4H), 1.96(s, 3H), 1.82-1.76(m, 1H), 1.52 1.44(m, 18H), 1.45-1.29(m, 1H), 1.07-1.04(m, 6H). MS: (M+H)+=456, (M+Na)+=478, (M-H)-=454, (M+35)=490. -393- WO 99/54299 PCT/US99/07945 AcHN, OH
H
0 >N HCIO OH 185B (±)-(2R.3S,5R,1'S)-2-(1-Acetamido-2-(N-isopropyI-N-hydroxyamino))ethyl 3-vinyl-pyrrolidine-5-carboxylic Acid Hydrochloride Salt The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-(N-isopropyl-N-hydroxyamino))ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)(2R,3R,5R,1'S)-2-(1-acetamido-3-ethyl)pentyl-3 methoxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 3.0 mg, 87%). 1 H NMR (MeOD-d 3 ) 8 5.83-5.71(m, 1H), 5.40(d, J=17.3Hz, 1H), 5.24(d, J=10.2Hz, 1H), 4.48(dd, J=7.8, 10.2Hz, 1H), 3.88-3.59(m, 4H), 3.17-3.10(m, 1H), 2.67-2.58(m, 1H), 2.10-1.99(m, 1H), 2.09(s, 3H), 1.33-1.17(m, 1H), 1.38(d, J=6.4Hz, 6H). MS: (M+H)+=300, (M-H)=298, (2M-H)-=597 -394- WO 99/54299 PCT/US99/07945 Example 186
CH
3 AcHN. OH HH TFA (±)-(2R, 3S,5R, 1'R)-2-(1 -Acetamido-2-oxo-2-phenyl)ethyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-oxo-2-phenyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 5.9 mg, 100%). 1 H NMR (DMSO-d 6 ) 5 8.62 (d, J= 9.8Hz, 1H), 7.93 (m, 2H), 7.68 (m, 1H), 7.55 (t, J= 7.9Hz, 2H), 5.61 (m, 1H), 5.48 (m, 1H), 5.19 (m, 1H), 4.50 (m, 1H), 3.98 (t, J= 9.8Hz, 1H), 3.30 (m, 1H), 2.38 (m, 1H), 1.73 (m, 1H), 1.71 (s, 3H), 1.59 (m, 3H). MS: (M+H)Y= 331, (M+Na)+= 353, (M-H)- = 329. -395- WO 99/54299 PCT/US99/07945 Example 187 (±)-(2R,3S,5R, 1 'R,2'R)-2-(1 -Acetamido-2-methoxy-4-vinyl)butyl-3-(cis-propen-1 yl)-pyDvrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN. O t Bu HN -K Boc O
OCH
3 C 187A (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-methoxy-4 vinyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound is prepared according to the method described in Example 84A, substituting (±)-(2R,3S,5R, 1 'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester for (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester.
CH
3 AcHN - OH H 0
OCH
3 TFA 187B (±)-(2R,3S,5R,1 'R,2'R)-2-(1-Acetamido-2-methoxy-4-vinyl)butyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound is prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxy-4-vinyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester. -396- WO 99/54299 PCT/US99/07945 Example 188 (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-methoxy-4-vinyl)butyl-3-(cis-propen-1 yl)-pvrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN. O t Bu N HNOCd
OCH
3 188A (±)-(2R,3S.5R, 1 'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-methoxy-4 vinyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 84A, substituting (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen- 1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester for (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0044 g, 22%). MS: (M+H)*=481, (M-H)-=479. CH 3 O AcHN O "OH H
OCH
3 TFA 188B (±)-(2R.,3S,5R.,1'R,2'S)-2-(1-Acetamido-2-methoxy-4-vinyl)butl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxy-4-vinyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1-t-butoxycarbonyl-2-(1 -397- WO 99/54299 PCT/US99/07945 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0031 g, 100%). 1 H NMR (DMSO-d 6 ) 8 7.93 (d, J=9.2Hz, 1H), 5.81 (m, 1H), 5.49 (m, 1H), 5.26 (m, 1H), 5.1-4.9 (m, 2H), 4.29 (m 1H), 4.03 (m, 2H), 3.68 (m, 1H), 3.26 (m, 1H), 3.25 (s, 3H), 3.18 (quint., J=8.5Hz, 1H), 2.40 (dt, J=12.7,7.3Hz, 1H), 2.32 (M, 1H), 2.20 (m, 1H), 2.02 (m, 1H), 1.85 (s, 3H), 1.68 (m, 1H), 1.64 (m, 1H), 1.61 (dd, J=6.7,1.8Hz, 3H), 1.55-1.40 (m, 2H). MS: (M+H)+=325, (M+Na)+=347, (M-H)=323. Example 189 (±)-(2R,3R.5R,1'R,2'S)-2-(1 -Acetamido-2,3-dihydroxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt TBDPSO-, MsO H N OtBu
N
3 BOC 0 189A (±)-(2R. 3R.,5R,1'S)-l-t-Butoxycarbonyl-2-(1-methanesulfonyloxy-3 azido)ethyl-3-t-butvldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 123J substituting (±)-(2R,3R,5R, 1 'S)-l-t-butoxycarbonyl-2-oxiranyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (2R,3S,5R,1'S)-2-oxiranyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 9.0 g, 90%). -398- WO 99/54299 PCT/US99/07945 1 H NMR (DMSO-d 6 ) (rotamers) 6 7.62-7.58 (m, 4H), 7.49-7.38 (m 6H), 4.97-4.79 (m, 1H), 4.19-4.02 (m, 2H), 3.79-3.48 (m, 2H), 3.15 and 3.13 (2s, 3H), 2.49-2.39 (m, 2H), 1.98-1.74 (m, 1H), 1.43-1.25 (m, 18H), 1.02 and 1.00 (2s, 9H) MS: (M+H)*= 703, (M+Na)*= 725 TBDPSO-', Hn OtBu 189B (±)-(2R. 3R,5R, l'S)-1 -t-butoxycarbonyl-2-aziridinyl-3-t butyldiphenylsilviyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester The title compound was prepared according to the method described in Example 123K substituting(±)-(2R,3R,5R, l'S)-1 -t-butoxycarbonyl-2-(1 methanesulfonyloxy-3-azido)ethyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'S)-1-t-butoxycarbonyl-2 (1-methanesulfonyloxy-3-azido)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester(yield: 5.9 g, 79%). 1 H NMR (DMSO-de) (rotamers) 8 7.60-7.56 (m, 4H), 7.49-7.39 (m 6H), 4.11-4.05 (m, 1H), 3.67-3.48 (m, 2H), 3.42-3.30 (m, 1H), 2.49-2.39 (m, 1H), 2.25 1.61 (m, 5H), 1.40, 1.35, 1.33, and 1.27 (4s, 18H), 0.99 and 0.98 (2s, 9H) MS: (M+H)*= 581, (M+Na) + = 603 -399- WO 99/54299 PCT/US99/07945 TBDPSO-, AcNOtBu 189C (±)-(2R, 3RR, 5R,1 'S)-l1-t-Butoxycarbonyl-2-N-acetylaziridinyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 123L substituting (±)-(2R,3R,5R,1'S)-1l-t-butoxycarbonyl-2-aziridinyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1 'S)-1 -t-butoxycarbonyl-2-aziridinyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 3.1 g, 96%). 1 H NMR (DMSO-d 6 ) (rotamers) 8 7.60-7.57 (m, 4H), 7.49-7.39 (m 6H), 4.18-4.11 (m, 1H), 3.71-3.51 (m, 3H), 2.76-2.68 (m, 1H), 2.58-2.45 (m, 1H), 2.46 and 2.39 (2d, J=6.1, 6.1Hz, 1H), 2.40 and 2.47 (2m, 1H), 2.08 and 2.05 (2d, J=3.1, 3.1 Hz, 1H), 2.02 and 1.99 (2s, 3H), 1.94-1.79 (m, 1H), 1.41, 1.36, 1.35 and 1.29 (4s, 18H), 0.99 and 0.98 (2s, 9H) MS: (M+H) = 623, (M+Na) = 645 TBDPSO-, AcHN- Otau A H N-" Boc" I O B u AcO O 189D (±)-(2R,3R,5R, 1'R)-l1-t-Butoxycarbonyl-2-(1-acetamido-2-acetoxy)ethyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3R, 5R, 1 'S)-1 -t-butoxycarbonyl-2-N-acetylaziridinyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (2.75g, 4.40 mmole) was reacted with potassium acetate (2.49 g, 25.37 mmole) and acetic acid (1.45 mL, 25.37 mmole) in DMSO (45 mL) at 1000C for 16 hours. The -400- WO 99/54299 PCT/US99/07945 reaction was quenched with 1N NaHCO 3 (100 mL) and diluted with ethyl acetate (300 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 100% dichloromethane to 50% dichloromethane/ethyl acetate to provide the title compound (yield: 2.45g, 81%). MS: (M+H)*=683, (M+Na)*=705, (M-H)-=681, (M+CI)=717 TBDPSO-, AcHN- Bo ' OtBu HO B oc 0 189E (±)-(2R,3R,5R, 1 'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy)ethyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3R,5R,1'R)-l1-t-Butoxycarbonyl-2-(1-acetamido-2-acetoxy)ethyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (2.45 g, 3.58 mmole) was reacted with potassium carbonate (1.48 g, 10.73 mmole) in methanol (18 mL) and THF (18mL) at 25 0 C for 45 minutes. The reaction was quenched with water (100 mL) and diluted with ethyl acetate (200 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 85% dichloromethane/ethyl acetate to 100% ethyl acetate to provide the title compound (yield: 2.05 g, 90%). MS: (M+H) =641, (M+Na)*=663, (2M+Na+H)*=1304, (M-H)-=639, (M+CI) =675 -401- WO 99/54299 PCT/US99/07945 TBDPSO, AcHN- ' /OtBu H Boc O H 189F (±)-(2R. 3R,5R, 1'R)-1 -t-Butoxycarbonyl-2-(1-acetamido-2-formyl)ethyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 41A substituting (±)-(2R,3R,5R,1'R)-l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)ethyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R)-1l-t-butoxycarbonyl 2-(1-acetamido-2 hydroxy)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (DMSO-d 6 ) (rotamers) 9.49(d, J=16.3, 1H), 8.33 and 8.29(2d, J=8.8 and 8.8Hz, 1H), 7.58-7.38(m, 10 OH), 4.94 and 4.84(2dd, J=4.4, 8.8Hz and 4.4, 8.8Hz, 1H), 4.26-3.37(m, 4H), 2.47-2.30(m, 1H), 1.97-1.83(m, 1H), 1.92(s, 3H), 1.42-1.18(m, 18H), 1.42-1.18(m, 1H), 1.00-0.97(m, 9H). MS: (M+H)*=639, (M-H)=637 TBDPSO, AcHN- 'n f' OtBu H Boc o 189G (±)-(2R,3R,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-1l-vinyl)methyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 118A substituting (±)-(2R, 3R,5R, 1'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-formyl)ethyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R)-1-t-butoxycarbonyl 2-(1 -402- WO 99/54299 PCT/US99/07945 acetamido-2-formyl)ethyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (DMSO-d 6 ) (rotamers) 7.99-7.74(m, 1H), 7.59-7.39(m, 10H), 5.80-5.68(m, 1H), 5.21-5.01(m, 3H), 3.97-3.31(m, 1H), 3.78-3.74(m, 1H), 3.60 3.46(m, 2H), 2.53-2.37(m, 1H), 2.09-1.72(m, 1H), 1.87(s, 3H), 1.42-1.23(m, 19H), 1.00-0.99(m, 9H). TBDPSO- TBDPSO AcHN-HN" tB u AcHN-H-. OtBu BocO Boc O OH OH OH OH 189H (±)-(2R,3R,5R, 1 'R,2'R) and (±)-(2R,3R,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2 (1-acetamido-2,3-dihydroxy)propyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5 carboxylic Acid t-Butyl Ester The title compounds were prepared according to the method described in Example 20A substituting (±)-(2R,3R,5R,1'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-vinyl)ethyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R)-1l-benzyl-2-vinyl-3-t-butyldimethylsilyloxymethyl pyrrolidine-5-carboxylic acid t-butyl ester (±)-(2R,3R,5R, I1'R,2'S) isomer (yield: 311mg, 24%) (±)-(2R,3R,5R,1'R,2'R) isomer (yield: 700 mg, 54%). (±)-(2R,3R,5R, 1'R,2'S) 1 H NMR (DMSO-de) (rotamers) 7.62-7.39(m, 11H), 4.56 and 4.51(d, J=4.8, 1H), 4.46-4.39(m, 2H), 3.97-3.82(m, 1H), 3.74 3.47(m, 3H), 3.28-3.21(m, 2H), 2.89-2.64(m, 1H), 2.51-2.45(m, 1H), 2.05-1.8(m, 1H), 1.87-1.86(m, 3H), 1.43-1.23(m, 19H), 0.99-0.98(m, 9H). (±)-(2R,3R,5R,1'R,2'R) 1 H NMR (DMSO-d 6 ) (rotamers) 7.63-7.40(m, 11H), 4.56-4.54(d, J=4.8, 1H), 4.47-4.33(m, 2H), 3.94-3.80(m, 1H), 3.85-3.80(m, 1H), 3.76-3.68(m, 1H), 3.60-3.51(m, 1H), 3.44-3.35(m, 1H), 3.30-3.21(m, 1H), -403- WO 99/54299 PCT/US99/07945 2.78-2.62(m, 1H), 2.46-2.31(m, 1H), 2.07-1.98(m 1H), 1.83(s, 3H), 1.39-1.29(m, 19H), 1.00-0.99(m, 9H).
TBDPSO
AcHN- OtBu '0 Boc O 0 1891 (±)-(2R,3R,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-1l-(2,2 dimethyl-1,3-dioxolan-4-yl))methyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5 carboxylic Acid t-Butyl Ester (±)-(2R,3R,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2,3 dihydroxy)propyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t butyl ester was reacted with 2,2-dimethoxypropane (1.1 ml, 9.09 mmole) and p Toluenesulfonic acid (4.3 mg, 0.023 mmole) in tetrahydrofuran (4.5 mL) at 25 0 C for 45 minutes. The reaction was quenched with triethylamine (3 mL). Stirring was continued for an additional 10 minutes. The reaction was then diluted with 10% NaHCO 3 (15 mL) and extracted with ethyl acetate (45 ml). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was carried over to the next step. purified by chromatography on silica gel using 100% dichlormethane to 94% dichloromethane/methanol to provide the title compound (yield: 194 mg, 91%). -404- WO 99/54299 PCT/US99/07945 HO-, dimethyl-1,3-dioxolan-4-yl))methyl-3-hydroxymethyl-pyrrolidine-5-carboxylic Acid AcHN-t-Butyl Ester OBu The title compound was prepared according to the method described in '0 0 1 89J (±)-(2R. 3R. 5R. 1'R.2'S)-1-t-Butoxvcarbonl-2-(1 -acetamido-1 -(2.2 d i methyl- 1, 3-d ioxolan-4-yl)) methvl-3-hyd roxymethyl-pyrro lid i ne-5-ca rboxyl ic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 123G substituting (±)-(2R,3R,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-1l-(2,2-dimethyl-1,3-dioxolan-4-yl))methyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester for (±) (2R, 3 R,5R,1'S)-1 -t-butoxycarbonyl-2-oxiranyl-3-t-butyldiphenylsilyloxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. The resulting residue was purified by chromatography on silica gel using 100% dichlormethane to 94% dichloromethane/methanol to provide the title compound (yield: 194 mg, 91%). H O==I, AcHN- OtBu O 0 189JJ (±)-(2R, 3R,5R, 1 'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-1l-(2,2 dimethyl-1,3-dioxolan-4-yl))methyl-3-formyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 123H substituting (±)-(2R,3R,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-1 -(2,2-dimethyl-1,3-dioxolan-4-yl))methyl-3-hydroxymethyl-pyrrolidine -405- WO 99/54299 PCT/US99/07945 5-carboxylic acid t-butyl ester for (±)-(2R,3R,5R, 1'S)-1 -t-butoxycarbonyl-2 oxiranyl-3-hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester.
H
3 C AcHN- OBu HN Boc O 0 0--/< 189K (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-1l-(2,2-dimethyl 1,3-dioxolan-4-yl))methyl-3-(cis-propen-1-yl))-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 35A substituting (±)-(2R,3R,5R, 1 'R,2'S)-1 -t-butoxycarbonyl-2-(1 acetamido-1 -(2,2-dimethyl-1,3-dioxolan-4-yl))methyl-3-formyl-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1-t-butoxycarbonyl-2 (1-acetamido-3-methyl)butyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 11.5 mg, 59%). 1 H NMR (CDCI 3 ): 8 6.62 (d, 1H), 5.56 (m, 1H), 5.40 (m, 1H), 4.43 (m, 1H), 4.25 (m, 1H), 4.16 (m, 1H), 4.02 (m, 1H), 3.88 (m, 1H), 3.54 (m 1H), 3.14 (m, 1H), 2.54 (m 1H), 2.04 (s, 3H), 1.71 (m 1H), 1.60 (dd, 3H), 1.46 (s, 9H), 1.45 (s, 9H), 1.40 (s, 3H), 1.32 (s, 3H). MS: (M+H)*=483 -406- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH N HH * 0 OH OH TFA 189L (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2,3-dihydroxy)propyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-1 -(2,2-dimethyl- 1,3-dioxolan-4-yl))methyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic acid t-butyl ester in place (±)-(2R,3S,5R,1'R,2'S)-1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester. 1 H NMR (DMSO-d 6 ): 5 7.84 (d, J=9Hz, 1H), 5.49 (m, 1H), 5.27 (m, 1H), 4.47 (m, 1H), 4.25 (m, 1H), 4.17 (m, 1H), 3.75 (m, 1 H), 3.59 (m, 1H), 3.35 (m, 1H), 3.18 (m, 1H), 2.43 (m, 1H), 1.81 (s, 3H),1.55 (dd, 3H). MS: (M+H)+=287 -407- WO 99/54299 PCTIUS99/07945 Example 190 (±)-(2R,3R.5R, 1'R.2'R)-2-(1-Acetamido-2,3-dihydroxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
TBDPSO
HN Boc O 0 0 190A (±)-(2R.3R,5R, 1 'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-1-(2,2 dimethyl-1,3-dioxolan-4-vl))methyl-3-t-butyldiphenyilsilyloxymethyl-pyrrolidine-5 carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 1891 substituting (±)-(2R,3R,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2,3-dihydroxy)propyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'R,2'S)-1-t Butoxycarbonyl-2-(1 -acetamido-2,3-dihydroxy)propyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester. -408- WO 99/54299 PCT/US99/07945 HO, AcHN- OtBu N
OC
O 0 190B (±)-(2R,3R,5R, 1 'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-1l-(2,2 dimethyl-1,3-dioxolan-4-yl))methyl-3-hydroxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 123G substituting (±)-(2R,3R,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-1 -(2,2-dimethyl-1,3-dioxolan-4-yl))methyl-3-t butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester for () (2R,3R,5R, 'S)-1 -t-butoxycarbonyl-2-oxiranyl-3-t-butyldiphenylsilyloxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. H O=d, AcHN- OtBu N H BocO 0 0 190C (±)-(2R,3R,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-1l-(2,2 dimethyl-1,3-dioxolan-4-yl))methyl-3-formyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 123H substituting (±)-(2R,3R,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-1 -(2,2-dimethyl-1,3-dioxolan-4-yl))methyl-3-hydroxymethyl-pyrrolidine 5-carboxylic acid t-butyl ester for (±)-(2R,3R,5R, 1'S)-1 -t-butoxycarbonyl-2 oxiranyl-3-hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester. -409- WO 99/54299 PCT/US99/07945
H
3 C t AcHN- OtBu HBoc O 0 190D (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-1l-(2,2 dimethyl-1,3-dioxolan-4-yl))methyl-3-(cis-propen-1 -yl))-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 35A substituting (±)-(2R,3R,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-1 -(2,2-dimethyl-1,3-dioxolan-4-yl))methyl-3-formyl-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1-t-butoxycarbonyl-2 (1-acetamido-3-methyl)butyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 42 mg, 61%). 1 H NMR (CDCI 3 ): 8 7.88 (d, 1H), 5.52 (m, 1H), 5.34 (m, 1H), 4.33 (m, 1H), 4.21 (m, 1H), 3.96 (m, 2H), 3.83 (m, 1H), 3.60 (m, 1H), 3.40 (m, 1H), 2.53 (m, 1H), 1.98 (s, 3H), 1.66 (dd, 3H), 1.46 (s, 9H), 1.44 (s, 9H), 1.41 (s, 3H), 1.33 (s, 3H). MS: (M+H) =483 -410- WO 99/54299 PCT/US99/07945
CH
3 AcHN. H N OH ' N OH OH TFA 190E (±)-(2R,3S,5R, 1'R,2'R)-2-(1-Acetamido-2,3-dihydroxy)propyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1-t-butoxycarbonyl-2-(1 acetamido-1 -(2,2-dimethyl-1, 3-dioxolan-4-yl))methyl-3-(cis-propen-1 -yl)) pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, I1'R,2'S)-1-t butoxycarbonyl-2-(l- acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester. 'H NMR (DMSO-d 6 ): 6 7.98 (d, J=9Hz, 1H), 5.48 (m, 1H), 5.29 (m, 1H), 4.60 (m, 1H), 4.30 (m, 1H), 4.12 (m, 1 H), 3.76 (m, 1 H), 3.52 (m, 1H), 3.46 (m, 1H), 3.32 (m, 1H), 3.18 (m, 1H), 2.40 (m, 1H), 1.84 (s, 3H), 1.60 (dd, 3H). MS: (M+H)+=287 -411- WO 99/54299 PCT/US99/07945 Example 193 (±)-(2R, 3S, 5R, 1 'R,2'S)-2-(1 -Acetamido-2-ethoxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN,. OtBu Boc 193A (±)-(2R,3S,5R, 1'R,2'S)-1l-t-Butoxycarbonyl-2-(1 -acetamido-2 ethoxy)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 88A, substituting ethyl iodide for methyl iodide (yield: 3.6 mg, 28%). MS: (M+H) = 483, (M+Na) = 505, (M-H)- = 481.
H
3 C AcHN. . OH N H 0 O TFA 193B (±)-(2R,3S,5R. 1'R,2'S)-2-(1-Acetamido-2-ethoxy)pentyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R,1 'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-ethoxy)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester, ester (yield: 3.2 mg, 100%). -412- WO 99/54299 PCT/US99/07945 1 H NMR (DMSO-d 6 ) 5 7.92 (d, J= 9.2Hz, 1H), 5.47 (m, 1H), 5.25 (m, 1H), 4.25 (m, 2H), 3.70 (m, 1H), 3.52 (m, 1H), 3.33 (m, 2H), 3.18 (m, 1H), 2.39 (m, 1H), 1.85 (s, 3H), 1.66 (m, 1H), 1.61 (dd, J= 6.7, 1.8Hz, 3H), 1.56 (m, 1H), 1.37 (m, 1H), 1.28 (m, 2H), 1.13 (m, 3H), 0.86 (t, J= 7.3Hz, 3H). MS: (M+H)*= 327, (M+Na)*= 349, (M-H)- = 325. Example 194 (±)-(2R,3S,5R, 1'R.2'R)-2-(1 -Acetamido-2-ethoxy)pentyl-3-(cis-propen-1 -vl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 OtBu AcHN. OtBu 'N Id ,r Boc O O 194A (±)-(2R.,3S,5R, 1'R,2'R)-l-t-Butoxycarbonyl-2-(1-acetamido-2 ethoxy)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound is prepared according to the method described in Example 88A, substituting ethyl iodide for methyl iodide. -413- WO 99/54299 PCT/US99/07945
H
3 C AcHN OH 'O O 114: H110 TFA 194B (±)-(2R,3S,5R, 1 'R,2'R)-2-(1-Acetamido-2-ethoxy)pentyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound is prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-ethoxy)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. Example 195 (±)-(2R,3S,5R, 1'S)-2-(1 -Acetamido-2-hydroxy)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt AcHN OH H HO O TFA The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'S)-1 -t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 19.9 mg, 100%). -414- WO 99/54299 PCT/US99/07945 1 H NMR (DMSO-d 6 ) 6 7.80 (d, J= 8.8Hz, 1H), 5.76 (m, 1H), 5.23 (d, J= 17.1Hz, 1H), 5.15 (m, 1H), 4.31 (m, 1H), 4.03 (m, 1H), 3.62 (m, 1H), 3.53 (m, 2H), 2.79 (m, 1H), 2.42 (m, 1H), 1.90 (s, 3H), 1.85 (m, 1H). MS: (M+H) = 243, (M+Na) = 265, (M-H) = 241. Example 196 (±)-(2R,3S, 5R, 1 'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-dimethylphosphonyl)propyl 3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3
CH
3 AcHN. OBu AcHN. OtBu 'N N K OH BocO 0OH Boc O OH OH
O=P-OCH
3
O=P-OCH
3
OCH
3
OCH
3 196A (±)-(2R,3S,5R, 1 'R.2'S) and (±)-(2R,3S,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2 (1-acetamido-2-hydroxy-3-dimethylphosphonyl)propyl-3-(cis-propen-1 -ylv) pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1 'R)-1 -t-Butoxycarbonyl 2-(1 -acetamido-1 -formyl)methyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (78 mg, 0.19 mmol) in THF (5 mL) was added dropwise to a solution of dimethylphosphonylmethyl lithium (3M) (0.32 mL, 0.95 mmol) in THF (20 mL) at -78 0 C and reacted for 40 minutes. The reaction was quenched with water (10 mL) and saturated aqueous ammonium chloride (10 mL) followed by extraction using dichloromethane (2 x 50 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 5-10% methanol in dichloromethane to provide the title -415- WO 99/54299 PCT/US99/07945 compounds (±)-(2R,3S,5R,1'R,2'R) isomer (yield: 27 mg, 27%) and (+) (2R,3S,5R,1'R,2'S) isomer (yield: 5.5 mg, 6%). (±)-(2R,3S,5R,1'R,2'R) = 'H NMR (CDC 3 ) 8 5.98 (m, 1H), 5.58(m, 1H), 5.35(m, 1H), 4.94(m, 1H), 4.14(m, 2H), 3.74(m, 8H), 3.06(m, 1H), 2.64(m, 1H), 2.03(s, 3H), 1.95(m, 1H), 1.83(m, 3H), 1.53(s, 9H), 1.46(s, 9H) MS: (M+H)+=535, (M-H)- =533 (±)-(2R,3S,5R,1'R,2'S) MS: (M+H)Y=535, (M-H)-=533
CH
3 AcHN. OH N H 0 OH O=P-OCH3 TFA
OCH
3 196B (±)-(2R,3S,5R,1 'R,2'S)-2-(1-Acetamido-2-hydroxy-3 dimethylphosphonyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-dimethylphosphonyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine 5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)- 1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 3 mg, 96%). 'H NMR (DMSO-d6) 6 7.98(d, J=9.2 HZ, 1H), 5.48(M, 1H), 5.28(m, 1H), 4.36(m, 1H), 4.30(m, 1H), 4.08(m, 2H), 3.70(m, 2H), 3.60(m, 6H), 3.18(m, 1H), 2.40(m, 1H), 2.05(m, 1H), 1.85(s, 3H), 1.60(dd, J=6.2, 1.2 HZ, 3H) -416- WO 99/54299 PCT/US99/07945 MS: (M+H)+=379, (M-H) =377 Example 197
CH
3 AcHN. OH 'N H 0 OH "OH O O=P-OCH 3 TFA
OCH
3 (±)-(2R,3S,5R, 1 'R,2'R)-2-(1 -Acetamido-2-hydroxy-3-dimethylphosphonyl)propyl 3-(cis-propen-1 -vyl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-dimethylphosphonyl)propyl-3-(cis-propen-1 -yl)-pyrrolidine 5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 13 mg, 96%). 1 H NMR (DMSO-de) 6 7.72 (d, J=9.2 HZ, 1H), 5.48(m, 1H), 5.24(m, 1H), 4.44(m, 1H), 4.15(m, 2H), 3.62(m, 7H), 3.54(m, 1H), 3.15(m, 1H), 2.40(m, 1H), 1.95(m, 1H), 1.82(s, 3H), 1.72(m, 1H), 1.54(dd, J=6.7, 1.2 HZ, 3H) MS: (M+H)*= 379, (M-H) = 377 -417- WO 99/54299 PCT/US99/07945 Example 198 (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-3-hydroxy)propyl-3-(cis-propen-1-ylv) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 OtBu AcHN. Ou N H Boc CO
OCH
3 198A (±)-(2R.3S,5R, 1 'S)-1 -t-Butoxycarbonyl-2-(1-acetamido-1 -(cis and trans-2 methoxyvinyl))methyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R, 1'R)- 1 -t-Butoxycarbonyl-2-(1 -acetamido- 1 -formyl)methyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (113 mg, 0.28 mmol) was added to a solution of (methoxymethyl)triphenylphosphonium bromide (240 mg, 0.70 mmol) and potassium t-butoxide (0.56 mL, 0.56 mmol, 1M in THF) in toluene (3 mL) at 0 0 C for 15 minutes. The reaction was quenched with saturated aqueous ammonium chloride (3 mL) followed by extraction using dichloromethane (2 X 3 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/4: ethyl acetate/hexane to provide the title compounds 'H NMR (CDCl 3 ) 5 8.65 (brd, 1H) 6.01 (d, J=5.7Hz, 1H), 5.40 (m, 3H), 5.11 (brt, 1H), 4.15 (m, 2H), 3.72 (m, 1H) 3.61 (s, 3H), 3.00 (m, 1H), 2.42 (m, 1H), 1.94 (s, 3H), 1.64 (dd, J=1.4, 5.0Hz, 3H), 1.45 (m, 9H), 1.25 (m, 9H) MS: (M+H) = 439. -418- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OtBu N N O H- Boc O H 198B (±)-(2R,3S,5R, 1 'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-formyl)ethyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-(cis and trans-2 methoxyvinyl))methyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (21 mg, 0.048 mmol) was reacted with LiBr (37 mg, 0.43 mmol) and AG50W-X2 ion exchange resin in CH 3 CN (2 mL) and water (0.1 mL) at room temperature for 45 minutes. The reaction was filtered and quenched with saturated aqueous sodium bicarbonate (1 mL) followed by extraction using dichloromethane (2 X 1 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/4: ethyl acetate/hexane to provide the title compound. 1 H NMR (CDCI 3 ) 5 9.70 (dd, J=1.3, 2.4Hz, 1H), 8.11 (d, J=7.8Hz, 1H), 5.54 (m, 1H), 5.41 (t, J=5.8Hz, 1H), 4.52 (m, 1H), 4.13 (dd, J=4.4, 4.8Hz, 1H), 3.75 (dd, J=2.7, 3.1Hz, 1H), 2.86 (m, 1H), 2.47 (m, 3H), 1.99 (s, 3H), 1.63 (dd, J=1.6, 5.1Hz, 3H), 1.46 (s, 9H), 1.45 (m, 1H), 1.44 (s, 9H) MS: (M+H) = 425; (M-H)- = 423. -419- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OtBu 'N H BocO OH 1980 (±)-(2R.,3S,5R, 1'S)-l1-t-Butoxycarbonyl-2-(1l-acetamido-3-hydroxy)propyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R,1'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-formyl)ethyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (9 mg, 0.02 mmol) was reacted with sodium borohydride (1 mg, 0.02 mmol) in methanol (0.1 mL) at room temperature for 20 minutes. The reaction was quenched with saturated aqueous ammonium chloride (1 mL) followed by extraction using dichloromethane (2 X 1 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/4: ethyl acetate/hexane to provide the title compound. 'H NMR (CDC13) 5 8.45 (d, J=7.5Hz, 1H), 5.55 (m, 1H), 5.34 (t, J=7.8Hz, 1H), 4.20 (dd, J=3.0, 5.4Hz, 2H), 3.71 (d, J=6.1Hz, 1H), 3.62 (m, 1H), 3.50 (t, J=9.1Hz, 1H), 2.92 (m, 1H), 2.41 (m, 1H), 2.04 (s, 3H), 1.66 (dd, J=2.0, 5.1Hz, 3H), 1.62 (m, 1H), 1.47 (s, 9H), 1.45 (m, 1H), 1.43 (s, 9H), 1.22 (m, 2H) MS: (M+H) = 427; (M-H)- = 425. -420- WO 99/54299 PCT/US99/07945
CH
3 AcHN. N/OH N H0 0 OH TFA 198D (±)-(2R,3S, 5R, I 'S)-2-(l -Acetamido-3-hydroxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'S)-l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)propyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1 'R,2'S)- 1-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. ester (yield: 4.6 mg, 100%). 1 H NMR (DMSO-d 6 ) 69.25 (brs, 1H), 8.13 (d, J=7.3Hz, 1H), 5.52 (m, 1H), 5.28 (brt, 1H), 4.32 (brt, 1H), 4.22 (m, 1H), 3.49 (m, 4H), 3.18 (m, 1H), 2.40 (m, 1H), 1.90 (s, 3H), 1.73 (m, 1H), 1.63 (dd, J=1.8, 5.5Hz, 3H), 1.57 (m, 1H) MS: (M-H) = 269; (M+H)Y = 271. -421- WO 99/54299 PCT/US99/07945 Example 199 (±)-(2R,3S,5R, 1'S,3'S)-2-(1 -Acetamido-3-hydroxy)pentyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 3cN AcHN. OtBU
CH
3 OtBu B O H Boc o OH OH 199A (±)-(2R,3S,5R, 1'S,3'S) and (±)-(2R,3S,5R,1'R,3'R)-1l-t-Butoxycarbonyl-2 (1-acetamido-3-hydroxy)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxvlic Acid t Butyl Ester (±)-(2R,3S,5R, l'S)-l-t-Butoxycarbonyl-2-(1-acetamido-2-formyl)ethyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (26 mg, 0.061 mmol) was reacted with ethylmagnesium bromide (3.0 M) (0.122 mL, 0.367 mmol) in THF (4 mL) at room temperature for 30 minutes. The reaction was quenched with saturated aqueous ammonium chloride (10 mL) and water (10 mL) followed by extraction using ethyl acetate (3 X 25 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel using 1/1 ethyl acetate/hexane followed by 2/1 ethyl acetate/hexane to provide the title compounds (±)-(2R,3S,5R,1'S,2'S) (yield: 6.7 mg, 24%) and (±)-(2R,3S,5R,1'S,2'R) (yield: 6.8 mg, 24%). (±)-(2R,3S,5R,1'S,2'S) MS: (M+H)+=455, (M+Na) =477, (M-H)=453. (±)-(2R,3S,5R,1 'S,2'R) MS: (M+H) =455, (M+Na) =477, (M-H)-=453. -422- WO 99/54299 PCT/US99/07945
CH
3 AcHN. .OH HH O OH TFA 199B (±)-(2R,3S,5R. 1'S.,3'S)-2-(1-Acetamido-3-hydroxy)pentvl-3-(cis-propen-1 yl)-pyDvrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, l'S,3'S)-1l-t-butoxycarbonyl-2-(1 acetamido-3-hydroxy)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. ester (yield: 6.2 mg, 100%). 1 H NMR (DMSO-d 6 ) 5 9.20 (bs, 1H), 8.18 (d, J=7.3 Hz, 1H), 5.51 (m, 1H), 5.27 (m, 1H), 4.30 (m, 1H), 4.25 (m, 1H), 3.58 (m, 1H), 3.41 (m, 1H), 3.18 (m, 1H), 2.39 (m, 1H), 1.90 (s, 3H), 1.75 (m, 1H), 1.64 (dd, J=7.5,1.5Hz, 3H), 1.51 (M, 1H), 1.38 (m, 1H), 1.32 (m, 1H), 0.83 (t, J=7.3Hz, 3H). MS: (M+H) = 299, (M+Na) = 321, (M-H) = 297, (2M-H)=595. -423- WO 99/54299 PCT/US99/07945 Example 200 (±)-(2R, 3S, 5 R, 1'S, 3'R)-2-(1 -Acetamido-3-hyd roxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. H N H OH TFA The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1l'S,3'R)-1l-t-butoxycarbonyl-2-(1 acetamido-3-hydroxy)pentyl-3-(cis-propen- 1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. ester (yield: 6.5 mg, 100%). 'H NMR (DMSO-d 6 ) 5 9.25 (bs, 1H), 8.15 (d, J=7.3 Hz, 1H), 5.52 (m, 1H), 5.27 (m, 1H), 4.31 (m, 2H), 3.52 (m, 1H), 3.36 (m, 1H), 3.19 (quint., J=8.5Hz, 1H), 2.38 (m, 1H), 1.92 (s, 3H), 1.75 (m, 1H), 1.64 (dd, J=7.3,1.5Hz, 3H), 1.48 (m, 1H), 1.33 (m, 2H), 0.85 (t, J=7.3Hz, 3H). MS: (M+H) = 299, (M+Na) = 321, (M-H) = 297, (2M-H)-=595. Example 201 -424- WO 99/54299 PCT/US99/07945 (±)-(2R, 3S,5R, 1'R)-2-(1 -Acetamido-2-oxo-3,3-difluoro-3-vinvl)Drovpyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OH N HH 0 / O F F TFA The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1 Acetamido-2-oxo-3,3-difluoro-3-vinyl)propyl-3-(cis-propen- 1-yl)-pyrrolidine-5 carboxylic Acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 0.0050 g, 100%). 1 H NMR (DMSO-d 6 ) 8 8.67 (d, J=8.5Hz, 1H), 6.1-5.95 (m, 1H), 5.78 (dd, J=17.1, 2.4Hz, 1H), 5.71 (d, 11.0Hz, 1H), 5.45 (m, 1H), 5.12 (m, 1H), 4.94 (t, J=9.2Hz, 1H), 4.51 (dd, J=12.2,6.1Hz, 1H), 3.98 (m, 1H), 3.24 (m, 1H), 2.32 (m, 1H), 1.73 (s, 3H), 1.66 (q, J=11.9Hz, 1H), 1.57 (dd, J=6.7,1.8Hz, 3H). MS: (M+H) = 331, (M+H 2 0) =349, (M+Na) + = 353, (M-H)- = 329, (2M-H) =659. -425- WO 99/54299 PCT/US99/07945 Example 202 TBDPSO- TBDPSO AcHN-~ OtBu AcHN- OB NN Boc oH Boc o OH OH 202A (±)-(2R,3R,5R,1 I'R,2'S) and (±)-(2R,3R,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2 (1-acetamido-2-hydroxy)butyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5 carboxylic Acid t-Butyl Ester The title compounds were prepared according to the method described in Example 41 B substituting (±)-(2R,3R,5R, 1'R)-1l-t-butoxycarbonyl-2-(1 -acetamido 2-formyl)ethyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1 'R)-1l-t-butoxycarbonyl-2-(1 -acetamido-2 formyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester to provide (±)-(2R,3R,5R,1'R,2'S) isomer (yield: 370 mg, 17%) and (±) (2R,3R,5R,1'R,2'R) isomer (yield: 1.2 g, 55%). (±)-(2R,3R,5R,1'R,2'S) 1 H NMR(d 6 -DMSO) 8 7.4-7.65 (m, 10OH), 4.47 (d, 1H), 4.32 (m, 1H), 3.87 (m, 2H), 3.68 (m, 1H), 3.55 (m, 1H), 3.25 (m, 1H), 2.7 (m, 1H), 2.45 (m, 1H), 2.0 (m, 1H), 1.83 (d, 3H), 1.28-1.4 (m, 18H), 0.95 (d, 9H), 0.83 (dt, 3H) MS: (M-H)- = 667, (M+35) + = 703; (M+H) + = 669, (M+Na) + = 691 (±)-(2R,3R,5R,1'R,2'R) 1 H NMR(d 6 -DMSO) 5 7.4-7.65 (m, 10H), 4.40 (dd, 1H), 4.12-4.32 (m, 1H), 3.82-3.96 (m, 1H), 3.66 (m, 2H), 3.52 (t, 1H), 2.6-2.8 (m, 1H), 2.45 (m, 1H), 1.76-2.0 (m, 1H), 1.87 (d, 3H), 1.25-1.4 (m, 18H), 0.95 (d, 9H), 0.83 (dt, 3H). MS: (M-H)- = 667, (M+35) + = 703; (M+H) + = 669, (M+Na) + = 691 -426- WO 99/54299 PCT/US99/07945
TBDPSO
AcHN- OtBu H Boc O 0
K
0 202B (±)-(2R,3R,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethyloxy)butyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3R,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2 hydroxy)butyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (0.58 g,0.87 mmole) was reacted with methoxymethyl chloride (1.15 mL, 10.07 mmole) and diisopropylethylamine (3.5 mL, 20.1 mmole) in dichloromethane (1 mL) at room temperature for 5 hours. The reaction was quenched with saturated NH 4 CI (100 mL) and diluted with ethyl acetate (200 mL). The organic layer was washed with water, and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 5% methanol/methylene chloride to provide the title compound (yield: 0.64 g, 98%). 1 H NMR(d 6 -DMSO) 5 7.4-7.65 (m, 10H), 4.70 (s, 1H), 4.62 (s, 1H), 4.35 4.55 (m, 2H), 3.75-3.95 (m, 2H), 3.68 (m, 1H), 3.55 (m, 1H), 3.25 (m, 1H), 3.24 (s, 3H), 2.55 (m, 1H), 2.45 (m, 1H), 2.0 (m, 1H), 1.85 (s, 3H), 1.28-1.4 (m, 18H), 0.99 (d, 9H), 0.8 (dt, 3H) MS: (M-H)- = 755, (M+35) + = 791; (M+H) + = 757, (M+Na) + = 779 -427- WO 99/54299 PCT/US99/07945
HO
AcHN- OtBu HBo c 0 0 Lo
K
0 202C (±)-(2R,3R,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethyloxy)butyl-3-hydroxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 123G substituting (±)-(2R,3R,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxymethyloxy)butyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine 5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R, 1'S)-1 -t-butoxycarbonyl 2-oxiranyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.416 g, 95%). 1 H NMR(d 6 -DMSO) 8 7.45 (t, 1H), 4.62-4.74 (m, 3H), 4.48 (m, 1H), 3.85 (m, 2H), 3.55-3.6 (m, 2H), 3.45 (t, 1H), 3.2-3.4 (m, 2H), 3.25 (d, 3H), 2.4 (m, 2H), 1.82 (d, 3H), 1.58 (m, 3H), 1.32-1.45 (m, 18H), 0.82 (dt, 3H). MS: (M-H)- = 517, (M+35) + = 553; (M+H) + = 519, (M+Na) = 541 -428- WO 99/54299 PCT/US99/07945 H AcHN- OtBu N H Boc 0 202D (±)-(2R.3R,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethyloxy)butyl-3-formyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 123H substituting (±)-(2R,3R,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxymethyloxy)butyl-3-hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1-t-butoxycarbonyl-2-oxiranyl-3 hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.335 g, 80.8%). 1 H NMR(d 6 -DMSO) 8 9.55 (d, 1H), 7.48 (m, 1H), 4.55-4.72 (m, 4H) 3.9 (d, 1H), 3.6 (m, 2H), 3.45 (m, 3H), 3.32 (s, 3H), 3.05 (t, 1H), 2.25-2.45 (m, 4H), 1.83 (s, 3H), 1.58 (m, 3H), 1.30-1.45 (m, 18H), 0.86 (dt, 3H). MS: (M-H)- = 515, (M+35) + = 551; (M+H) + = 517 -429- WO 99/54299 PCT/US99/07945 OH AcHN- N OtBu HBoc O O OC 0 0 202E (±)-(2R,3R,5R, 1 'R,2'S,1"RS)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethyloxy)butyl-3-(1 -hydroxy-2-propyn-1 yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 38A substituting (±)-(2R,3R,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxymethyloxy)butyl-3-formyl-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3R,5R, 1'S)-1l-t-butoxycarbonyl-2-(1-acetamido-3 methyl)butyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.27 g, 83%). MS: (M-H)- = 541, (M+35) + = 577; (M+H) + = 543, (M+Na) = 565 0o AcHN- OtBu methoxymethloxy)butl-3-( 1-oxo-2-propyn-1 -yl)-pyrrol id ine-5-carboxylic Acid t Butyl Ester The title compound was prepared according to the method described in Example 38B substituting (+)-(2R,3R,5R,1'R,2'S,1"RS)-1-t-butoxycarbonyl-2-(1 -430- WO 99/54299 PCT/US99/07945 acetamido-2-methoxymethyloxy)butyl-3-(1-hydroxy-2-propyn-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S,1"RS)-1-t butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-(1 -hydroxy-2-propyn-1 -yl) pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.2 g, 74%). 1H NMR(d 6 -DMSO) 87.49 (br d, 1H), 5.0 (d, 1H), 4.7 (br s, 1H), 4.55-4.7 (m, 3H), 3.88 (br d, 1H), 3.5-3.7 (m, 2H), 3.43 (t, 2H), 3.2-3.4 (m, 2H), 3.24 (s, 3H), 2.4-2.7 (m, 2H), 1.84 (s, 3H), 1.5-1.7 (m, 2H), 1.30-1.45 (m, 18H), 0.86 (dt, 3H) MS: (M-H)- = 539, (M+35) + = 575; (M+H) + = 541, (M+Na)* = 563 AcHN- OtBu N H Boc 0
K
0 202G (±)-(2R.,3R,5R,1'R.2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethyloxy)butyl-3-(pyrazol-3-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 38C substituting (±)-(2R,3R,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxymethyloxy)butyl-3-(1-oxo-2-propyn-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1-t-butoxycarbonyl-2 (1-acetamido-3-methyl)butyl-3-(1-oxo-2-propyn-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 180 mg, 87%). -431- WO 99/54299 PCT/US99/07945 1 H NMR(d 6 -DMSO) 6 7.57 (brt, 2H), 6.1 (d, 1H), 4.50-4.7 (m, 4H) 3.95 (m, 1H), 3.4-3.6 (m, 3H), 3.3-3.4(m, 3H), 3.22 (d, 3H), 2.55-2.65 (m, 1H), 2.2 (m, 1H), 1.85 (s, 3H), 1.5-1.7 (m, 2H), 1.15-1.45 (m, 18H), 0.86 (dt, 3H). MS: (M-H)- = 553, (M+35) + = 589; (M+H) + = 553, (M+Na)* = 577 HN
H
OH TFA 202H (±)-(2R,3R,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)butyl-3-(pyrazol-3-yvl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3R,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxymethyloxy)butyl-3-(pyrazol-3-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place (±)-(2R,3R,5R, l'S)-2-(1-acetamido-3-ethyl)pentyl-3 methoxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester. Chromatography on silica gel with 2-propanol:acetic acid:ethyl acetate:water 1:1:3:1 followed by the addition of 0.1% trifluoroacetic acid gave the title compound (yield: 15 mg, 55%). 1 H NMR(d 6 -DMSO) 6 7.95 (d, 1H), 7.65 (brs, 1H), 6,18 (d, 1H), 4.37 (m, 1H), 4.23 (m, 1H), 4.38 (m, 1H), 4.56 (m, 1H), 2.63 (m, 1H), 2.10 (m, 1H), 1.78 (s, 3H), 1.50 (m, 1H), 1.25 (m, 1H), 0.83 (t, J=7.46 Hz, 3H). MS: (M-H)- = 309, (M+35) + = 345; (M+H) + = 311, (M+Na) + = 333 -432- WO 99/54299 PCT/US99/07945 Example 203 (±)-(2R.3R,5R, 1'R,2'R)-2-( 1-Acetamido-2-hydroxy)butyl-3-(pyrazol-3-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
TBDPSO
AcHN- OtBu 'N Boc 0 203B (±)-(2R,3R,5R, 1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethyloxy)butyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compounds were prepared according to the method described in Example 202B substituting (±)-(2R,3R,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'R,2'S)-1-t butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-t-butyldiphenylsilyloxymethyl pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.217 g, 96%). 1 H NMR(d 6 -DMSO) 67.4-7.65 (m, 10H), 4.70 (s, 1H), 4.62 (s, 1H), 4.35 4.55 (m, 2H), 3.75-3.95 (m, 2H), 3.68 (m, 1H), 3.55 (m, 1H), 3.25 (m, 1H), 3.24 (s, 3H), 2.55 (m, 1H), 2.45 (m, 1H), 2.0 (m, 1H), 1.85 (s, 3H), 1.28-1.4 (m, 18H), 0.99 (d, 9H), 0.8 (dt, 3H). MS: (M-H)- = 755, (M+35) + = 791; (M+H) + = 757, (M+Na) + = 779 -433- WO 99/54299 PCT/US99/07945 HO-, AcHN- OtBu 'N OC 0
K
0 203C (±)-(2R,3R,5R, 1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethyloxy)buyl-3-hydroxvmethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 123G substituting (±)-(2R,3R,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxymethyloxy)butyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine 5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1-t-butoxycarbonyl 2-oxiranyl-3-t-butyldiphenylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.124'g, 83%). 1H NMR(d 6 -DMSO) 6 7.42 (dd, 1H), 4.62-4.8 (m, 3H), 4.48 (m, 1H), 3.6 3.85 (m, 3H), 3.35-3.6 (m, 4H), 3.25 (s, 3H), 2.25 (m, 1H), 2.4 (m, 1H), 2.28 (m, 1H), 1.82 (s, 3H), 1.58 (m, 3H), 1.32-1.45 (m, 18H), 0.9 (dt, 3H). MS: (M-H) = 517, (M+35) + = 553; (M+H) + = 519, (M+Na)* = 541 -434- WO 99/54299 PCT/US99/07945 H O==d AcHN- OtBu 'N H Boc o 0 0 203D (±)-(2R,3R,5R, 1 'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethyloxy)butyl-3-formvl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 123H substituting (±)-(2R,3R,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxymethyloxy)butyl-3-hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1l-t-butoxycarbonyl-2-oxiranyl-3 hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.106 g, 86%). 1 H NMR(d 6 -DMSO) 89.58 (d, 1H), 7.58 (dd, 1H), 4.6-4.72 (m, 3H), 4.48(d, 1H), 3.88 (d, 1H), 3.4-3.65 (m, 5H), 3.24 (s, 3H), 3.15 (dd, 1H), 2.20-2.48 (m, 4H), 1.86 (s, 3H), 1.58 (m, 3H), 1.30-1.40 (m, 18H), 0.86 (t, 3H). MS: (M-H)- = 515, (M+35) + = 551; (M+H) + = 517 -435- WO 99/54299 PCT/US99/07945 OH AcHN- OtBu HN Boc O 0 1 203E (±)-(2R,3R,5R,1'R,2'R,1"RS)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethyloxy)butyl-3-(1-hydroxy-2-propyn-1-yI)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 38A substituting (±)-(2R,3R,5R, 1 'R,2'R)-1l-t-butoxycarbonyl-2-( 1 acetamido-2-methoxymethyloxy)butyl-3-formyl-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3R,5R,1'S,1"RS)-1l-t-butoxycarbonyl-2-(1 acetamido-3-methyl)butyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 32 mg, 76%). MS: (M-H) = 541, (M+35) + = 577; (M+H) + = 543, (M+Na) = 565 0o AcHN- N OtBu BOC 0 "O 203F (±)-(2R,3R,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethyloxy)butyl-3-(1-oxo-2-propyn-1 -vil)-pyrrolidine-5-carboxylic Acid t Butyl Ester The title compound was prepared according to the method described in Example 38B substituting (±)-(2R,3R,5R,1'R,2'R)-1l-t-butoxycarbonyl-2-(1 -436- WO 99/54299 PCT/US99/07945 acetamido-2-methoxymethyloxy)butyl-3-(1-hydroxy-2-propyn-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S,1"RS)-1-t butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-(1-oxo-2-propyn-1 -yl) pyrrolidine-5-carboxylic acid t-butyl ester (yield: 25 mg, 78%). 1H NMR(d 6 -DMSO) 6 7.49 (br d, 1H), 5.0 (d, 1 H), 4.7 (br s, 1H), 4.55-4.7 (m, 3H) 3.88 (br d, 1H), 3.5-3.7 (m, 2H), 3.43 (t, 2H), 3.2-3.4 (m, 2H), 3.24 (s, 3H), 2.4-2.7 (m, 2H), 1.84 (s, 3H), 1.5-1.7 (m, 2H), 1.30-1.45 (m, 18H), 0.86 (dt, 3H). MS: (M-H)- = 539, (M+35) + = 575; (M+H) + = 541, (M+Na) = 563 HN!N' AcHN-. OtBu 'N H Boc I 203G (±)-(2R,3R,5R,1'R,2'R)-1l-t-Butoxycarbonyl-2-(1-acetamido-2 methoxymethvyloxy)butyl-3-(pyrazol-3-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 38C substituting (±)-(2R,3R,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxymethyloxy)butyl-3-(1-oxo-2-propyn-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1-t-butoxycarbonyl-2 (1-acetamido-3-methyl)butyl-3-(1-oxo-2-propyn-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 18 mg, 72%). -437- WO 99/54299 PCT/US99/07945 1 H NMR(d 6 -DMSO) 8 7.57 (m, 2H), 6.1 (d, 1H), 4.40-4.7 (m, 4H) 3.93 (m, 1H), 3.4-3.6 (m, 3H), 3.3-3.4(m, 3H), 3.22 (d, 3H), 2.55-2.65 (m, 1H), 2.2 (m, 1H), 1.85 (s, 3H), 1.5-1.7 (m, 2H), 1.15-1.45 (m, 18H), 0.86 (m, 3H). MS: (M-H)- = 553, (M+35) + = 589; (M+H) + = 553, (M+Na) = 577 HN AcHN. OH N H 7 rn 0 OH TFA 203H (±)-(2R,3R,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxy)butyl-3-(pyrazol-3-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 15B, substituting (±)-(2R,3R,5R, 1'R,2'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxymethyloxy)butyl-3-(pyrazol-3-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-l-benzyl-2-(1-acetamido-3 ethyl)pentyl-3-(imidazol-2-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. Chromatography on silica gel with 2-propanol:acetic acid:ethyl acetate:water 1:1:3:1 followed by the addition of 0.1% trifluoroacetic acid gave the title compound (yield: 4 mg, 45%). 1 H NMR(d 6 -DMSO) 8 7.65 (d, 1H), 7.64 (d, 1H), 6,16 (d, 1H), 4.37 (m, 1H), 4.23 (m, 1H), 4.38 (m, 1H), 4.56 (m, 1H), 2.63 (m, 1H), 2.10 (m, 1H), 1.74 (s, 3H), 1.25-1.40 (m, 2H), 0.83 (t, J=7.46 Hz, 3H). MS: (M-H)- = 309, (M+35) + = 345; (M+H) + = 311, (M+Na) 4 = 333 -438- WO 99/54299 PCT/US99/07945 Example 204 (±)-(2R,3R,5R)-2-Acetamidomethyl-3-methoxycarbonyl-pyrrolidine-5-carboxylic Acid Hydrochloride.
TBDMSO
H
2 N OtBu H N 0 Ph 204A (±)-(2R,3R,5R)-1l-Benzyl-2-aminomethyl-3-t-butvldimethylsilyvloxymethyl pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound is prepared according to the method described in Example 1F, substituting (±)-(2R,3R,5R)-l1-benzyl-2-formyl-3-t butyldimethylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R, 3R,5R)-1-benzyl-2-(l-oxo-3-ethyl)pentyl-3-t-butyldimethylsilyloxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. MS: (M+H) = 435 TBDMSO-, AcNH OtBu N d Ph 204B (±)-(2R,3R,5R)-1l-Benzyl-2-acetamidomethyl-3-t butlidimethylsilyloxymethyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound is prepared according to the method described in Example 1G, substituting (±)-(2R,3R,5R)-1l-benzyl-2-aminomethyl-3-t butyldimethylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'R)- and (±)-(2R,3R,5R, 1'S)-1-benzyl-2-(1-amino-3-ethyl)pentyl-3 t-butyldimethylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester. -439- WO 99/54299 PCT/US99/07945 1 H NMR (CDCI 3 ): 8 7.2-7.35 (m, 5H), 6.14 (br, 1H), 3.86 (dd, J=18Hz, 13.5Hz, 2H), 3.67 (m, 1H), 3.60 (m, 1H), 3.49 (m, 1H), 3.28 (m, 1H), 3.06 (m, 1H), 2.19 (m, 2H), 1.95 (s, 3H), 1.45 (s, 9H), 0.91 (s, 9H), 0.07 (s, 6H). MS: (M+H) = 477 HO, AcNH OtBu HN d o Ph 204C (±)-(2R,3R,5R)-1l-Benzyl-2-acetamidomethyl-3-hydroxymethyl-pyrrolidine 5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 1H, substituting (±)-(2R,3R,5R)-1 -benzyl-2-acetamidomethyl-3-t butyldimethylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-t butyldimethylsilyloxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester. O
H
A
, H AcNH- OtBu N d Ph 204D (±)-(2R,3R. 5R)-1 -Benzyl-2-acetamidomethyl-3-formyl-pyrrolidine-5 carboxLylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 2A, substituting (±)-(2R,3R,5R)-1l-benzyl-2-acetamidomethyl-3 hydroxymethyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R,1'S)-1 -benzyl-2-(1-acetamido-3-ethyl)pentyl-3-hydrxoxymethyl pyrrolidine-5-carboxylic acid t-butyl ester. -440- WO 99/54299 PCT/US99/07945 1 H NMR (CDCI 3 ): 6 9.70 (s, 1H), 7.22-7.36 (m 5H), 5.82 (br, 1H), 3.83 (dd, J= 3.3Hz, 13.5Hz, 2H), 3.74 (m, 1H), 3.56 (d, J= 9Hz, 1H), 3.15 (m, 1H), 2.73 (m, 1H), 2.36-2.10 (m, 2H), 1.98 (s, 3H), 1.45 (s, 9H). MS: (M+H) = 361 O
CH
3 O0 AcNH OtBu HN d o Ph 204E (±)-(2R. 3R,5R)-1l-Benzyl-2-acetamidomethvl-3-methoxycarbonyl pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 2B and 2C, substituting (±)-(2R,3R,5R)-1l-benzyl -2-acetamidomethyl-3 formyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-1 benzyl-2-(1-acetamido-3-ethyl)pentyl-3-formyl-pyrrolidine-5-carboxylic acid t-butyl ester. 1 H NMR (CDCI 3 ): 6 7.45-7.20 (m, 5H), 5.96 (br, 1H), 3.90-3.73 (m, 4H), 3.71 (s, 3H), 3.52 (dd, J=9Hz, 2Hz, 1H), 3.13 (m, 1H), 2.84 (m, 1H), 2.36 (m, 1H), 2.18 (m, 1H), 1.97 (s, 3H), 1.45 (s, 9H). MS: (M+H) = 391 -441- WO 99/54299 PCT/US99/07945 O
CH
3 O0 AcNH OtBu N H 204F (±)-(2R,3R,5R)-2-Acetamidomethyl-3-methoxycarbonyl-pyrrolidine-5 carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 2D, substituting (±)-(2R,3R,5R)-1l-benzyl-2-acetamidomethyl-3 methoxycarbonyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R,1'S)-l-benzyl-2-(1-acetamido-3-ethyl)pentyl-3-methoxycarbonyl pyrrolidine-5-carboxylic acid t-butyl ester. 'H NMR (CDCI 3 ): 5 6.19 (br, 1H), 3.72 (m, 2H), 3.70 (s, 3H), 3.43 (m, 1H), 3.28 (m, 1H), 2.74 (m, 1H), 2.44 (m 1H), 2.21 (m, 1H), 2.00 (s, 3H), 1.48 (s, 9H). MS: (M+H) = 301 0
CH
3 O0 AcNH dOH N H HCI 204G (±)-(2R,3R.,5R)-2-Acetamidomethyl-3-methoxycarbonyl-pyrrolidine-5 carboxylic Acid Hydrochloride. The title compound was prepared according to the method described in Example 2E substituting (±)-(2R,3R,5R)-2-acetamidomethyl-3-methoxycarbonyl pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3R,5R,1'S)-2-(1 acetamido-3-ethyl)pentyl-3-methoxycarbonyl-pyrrolidine-5-carboxylic acid t-butyl ester. -442- WO 99/54299 PCT/US99/07945 1 H NMR (D 2 0): 6 4.42 (t, J=8.25Hz, 1H), 4.22 (m, 1H), 3.83 (m, 1H), 3.75 (s, 3H), 3.70-3.60 (m, 2H), 3.26 (m, 1H), 2.78 (m, 1H), 2.43 (m, 1H), 2.03 (s, 3H). MS: (M+H) = 245 Examples 205-213 HO- R AcHN OtBu , AcH N 11..n -KOH AcN N dN~H Boc O H O Example 204C The following title compounds were prepared according to the methods described in Examples 1-39 from the common intermediate prepared as described in Example 204C. Example 205 AcHN .. OH ~N H0 TFA (±)-(2R,3R,5R)-2-Acetamidomethyl-3-ethoxycarbonyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. 1 H NMR (D 2 0) 8 4.30 (t, J=8.2Hz, 1H), 4.21 (m, 3H), 3.62 (dd, J=2.4, 3.4Hz, 2H), 3.23 (m, 1H), 2.74 (m, 1H), 2.38 (m, 1H), 2.02 (s, 3H), 1.26 (m, 3H) MS: (M+H)*= 259; (M-H) = 257. -443- WO 99/54299 PCT/US99/07945 Example 206 "N AcHN g OH H HCI (±)-(2R,3R,5R)-2-Acetamidomethyl-3-(imidazol-2-yi)-pyrrolidine-5-carboxylic Acid Hydrochloride 'H NMR (D 2 0): 8 7.46 (s, 2H), 4.53 (dd, J=9.5Hz, J=8.5 Hz, 1H), 4.28 (m, 1H), 3.96 (m, 1H), 3.65 (m, 2H), 3.03 (dt, J=13.5Hz, J=7.6Hz, 1H), 2.46 (m, 1H), 1.94 (s, 3H). MS: (M+H) =253, (M-H)=251 Example 207 AcHN ,, OH ~N H0 TFA (±)-(2R,3S,5R)-2-Acetamidomethyl-3-vinyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. 'H NMR (D20) 6 5.74 (m, 1H), 5.24 (m, 2H), 4.20 (dd, J=1.7, 8.1Hz, 1H), 3.65 (m, 2H), 3.50 (m, 1H), 2.84 (m, 1H), 2.61 (m, 1H), 2.03 (s, 3H), 1.95 (m, 1H) -444- WO 99/54299 PCT/US99/07945 MS: (M+H) = 213. Example 208
H
3 C
H
3 C O AcHN ' N H0 TFA (±)-(2R,3R,5 R)-2-Acetamidomethyl-3-(2,2-dimethyl-vinyl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. 1 H NMR (D 2 0) 8 5.01 (br d, 1H), 4.18 (dd, J=2.1, 8.1Hz, 1H), 3.53 (m, 3H), 3.04 (m, 1H), 2.55 (m, 1H), 2.0 (s, 3H), 1.75 (m, 1H), 1.72 (s, 3H), 1.67 (s, 3H) MS: (M+H) = 241, (M+Na)* = 263; (M-H)- = 239. Example 209
H
3 CN O
H
3 C H OH H' H Io AcHNHH O HCI (±)-(2R,3R,5R)-2-Acetamidomethyl-3-(N,N-dimethylcarbamoyl)-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt. 1 H NMR (D 2 0) 6 4.60 (t, J=8.4Hz, 1H), 4.23 (m, 1H), 3.56 (d, J=5.8Hz, 2H) 3.50 (m, 1H), 3.10 (s, 3H), 2.94 (s, 3H), 2.88 (m, 1H), 2.19 (m, 1H), 2.00 (s, 3H) MS: (M+H) = 258, (M-H)- = 256. -445- WO 99/54299 PCT/US99/07945 Example 210
H
3 C O HN OH AcHN H O TFA (±)-(2R,3R,5R)-2-Acetamidomethyl-3-(N-methylcarbamoyl)-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt. 1 H NMR (D 2 0) 4.49 (t, J=8.5Hz, 1H), 4.10 (m, 1H), 3.57 (d, J=5.8Hz, 2H), 3.03 (m, 1H), 2.76 (m, 1H), 2.74 (s, 3H), 2.29 (m, 1H), 2.00 (s, 3H) MS: (M+H) = 244. Example 211
H
3 C o OH f N AcHN H O TFA (±)-(2R,3R,5R)-2-Acetamidomethyl-3-propionyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. H NMR (D 2 0) 8 4.24 (m, 2H), 3.55 (d, J=4.7Hz, 1H), 3.40 (m, 1H), 2.85 (m, 1H), 2.64 (m, 3H), 2.16 (m, 1H), 2.01 (s, 3H), 1.02 (t, J=7.1Hz, 3H) MS: (M+H) = 243; (M-H) = 241. -446- WO 99/54299 PCT/US99/07945 Example 212
H
3 C OH AcHN H O HCI (±)-(2R,3R,5R)-2-Acetamidomethyl-3-methoxymethyl-pyrrolidine-5-carboxylic Acid Hydrochloride 1 H NMR (D 2 0): 6 4.44 (t, J=6Hz, 2H), 3.77 (m, 1H), 3.65-3.48 (m, 3H), 3.35 (s, 3H), 2.64 (m, 1H), 2.56 (m, 1H), 2.03 (s, 3H), 2.00 (m, 1H). MS: (M+H) += 231, (M-H)- = 229 Example 213
H
3 C, OH N AcHN H O TFA (±)-(2R,3S,5R)-2-Acetamidomethyl-3-methyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. 1 H NMR (D 2 0) 8 4.30 (m, 1H), 3.64 (m, 1H), 3.48 (m, 1H), 3.20 (m, 1H), 2.64 (m, 1H), 2.03 (s, 3H), 1.76 (m, 1H), 1.32 (brt, 1H), 1.12 (m, 4H) MS: (M+H) = 201, (M+Na) = 223. -447- WO 99/54299 PCT/US99/07945 Example 214 (±)-(2R,3S,5R. 1'R)-2-(1 -Acetamido-2-ethylthio)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt BocHN', o N OtBu /B s 0 214A (±)-(2R.3S,5R,1'R)-l-t-Butoxvcarbonyl-2-(1-t-butoxycarbonylamino-2 ethylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. To a solution of ethanethiol (0.047 mL, 0.63 mmol) in THF (2 mL) at 0OC was added 2.5 M n-BuLi/hexane (0.248 mL, 0.62 mmol). The reaction mixture was stirred for 45 minutes and a solution of (±)-(2R,3S,5R,1'S)-1-t butoxycarbony-2-(N-t-butoxycarbonylaziridinyl)-3-vinyl-pyrro lidine-5-carboxylic acid t-butyl ester (0.08 g, 0.182 mmole) in THF (0.5 mL) was added followed by DMF (1.5 mL) and stirred at room temperature for 2 hours. The reaction was quenched with 1N NaHCO 3 (10 mL) and diluted with ethyl acetate (20 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 10% ethyl acetate/hexanes to provide the title compound (yield: 61 mg, 67%). 1H NMR(d 6 -DMSO) 5 6.74 (br d, 1H), 5.85 (m, 1H), 4.9-5.0 (m, 2H), 4.20 (m, 1H), 3.95 (m, 1H), 3.75 (d, 1H), 2.8-3.0 (dd, 1H), 2.5 (m, 3H), 1.65 (m, 1H), 1.32-1.45 (m, 27H), 1.17 (dt, 3H). MS: (M-H)- = 499; (M+H) + = 501, (M+Na) = 523 -448- WO 99/54299 PCT/US99/07945 Ac BocN. OtBu 214B (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-N-t-butoxycarbonylacetamido 2-ethylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1'R)-1l-t-Butoxycarbonyl-2-(1-N-t-butoxycarbonylamino-2 ethylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (58 mg, 0.116 mmole) was reacted with lithium hexamethyldisilazide (1 M) (1.16 mL, 1.16 mmole) in THF (3 mL) at -78 'C. After 0.5 hour at -78 oC and 1 hour at -40 oC, the above reaction mixture was reacted with acetyl chloride (0.166 mL, 2.33 mmole) at -30 oC for 0.3 hours. The reaction was quenched with 1N NaHCO 3 (10 mL) and extracted with ethyl acetate (20 mL). The organic layer was washed with water,and brine, dried over MgSO 4 , filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel using 10% ethyl acetate/hexanes to provide the title compound (yield: 28 mg, 44%). 1 H NMR(d 6 -DMSO) 5 5.88 (m, 1H), 4.9-5.0 (m, 2H), 4.52 (m, 1H), 4.33 (m, 1H), 4.1 (m, 1H), 2.78 (dd, 1H), 2.3-2.5 (m, 6H), 1.7 (m, 1H), 1.32-1.5 (m, 27H), 1.11 (t, 3H). MS: (M+H) + = 543. -449- WO 99/54299 PCT/US99/07945 AcHN, OH N S H TFA 2140 (±)-(2R,3R,5R,1'R)-2-(1-Acetamido-2-ethylthio)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R,1'R)-1l-t-butoxycarbonyl-2-(1-N-t butoxycarbonylacetamido-2-ethylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 7 mg, 95%). 1 H NMR(d 6 -DMSO) 8 8.15 (d, 1H), 5.72 (m, 1H), 5.05-5.2 (m, 2H), 4.2 4.4 (m, 2H), 4.33 (m, 1H), 2.93 (m, 1H), 2.7-2.8 (2d, 1H), 2.3-2.6(m, 3H), 1.85 1.95 (m, 1H), 1.93 (s, 3H), 1.17 (t, J=7.46 Hz, 3H) MS: (M+H) + = 287. -450- WO 99/54299 PCT/US99/07945 Example 215 (±)-(2R,3S,5R, 1'R, 3'S)-2-(1 -Acetamido-2-ethylsulfinyl)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt Ac Ac BocN. OtB BocN OtBu A O O 215A (±)-(2R,3S,5R, 1 'R,3'S) and (±)-(2R,3S,5R,1 'R,3'R)-1-t-Butoxycarbonyl-2 (1-N-t-butoxycarbonylacetamido-2-ethylsulfinyl)ethyl-3-vinyl-pyrrolidine-5 carboxvlic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1'R)-1 -t-Butoxycarbonyl-2-(1 -N-t-butoxycarbonylacetamido 2-ethylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (72 mg, 0.132 mmole) was reacted with 55% m-chloroperoxybenzoic acid (41 mg, 0.132 mmole) in CHCI 3 (1.5 mL) at -40 oC for 30 minutes. The reaction was concentrated in vacuo. The residue was purified by chromatography on silica gel using ethyl acetate to provide the title compounds (±)-(2R,3S,5R,1'R,3'S) isomer (yield: 14 mg, 18.9%) and (±)-(2R,3S,5R,1'R,3'R) (yield: 45 mg, 60.7%). (2R,3S,5R,1'R,3'S) 1 H NMR(d 6 -DMSO) 8 5.88 (m, 1H), 4.9-5.0 (m, 2H), 4.50 (m, 1H), 4.0-4.15 (m, 1H), 2.7-2.9 (m, 3H), 2.55 (m, 1H), 2.37 (s, 3H), 1.7 (m, 1H), 1.32-1.5 (m, 27H), 1.12 (t, 3H) MS: (M+H) + = 559, (M+Na) + = 581 -451- WO 99/54299 PCT/US99/07945 (2R,3S,5R,1'R,3'R) 1 H NMR(d 6 -DMSO) 6 5.88 (m, 1H), 4.9-5.0 (m, 2H), 4.50 (m, 1H), 4.03-4.15 (m, 1H), 3.2 (m, 1H) 3.1 (dd, 1H), 2.5-2.7(m, 2H), 2.38 (s, 3H), 1.75 (m, 1H), 1.32-1.5 (m, 27H), 1.12 (t, 3H) MS: (M+H) + = 559, (M+Na) + = 581 AcHN OH S 0 O TFA 215B (±)-(2R,3S,.5R,1'R,3'S)-2-(1-Acetamido-2-ethylsulfinyl)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R,3'S)-1l-t-butoxycarbonyl-2-(1-N-t butoxycarbonylacetamido-2-ethylsulfinyl)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. ester (yield: 9 mg, 86%). 1 H NMR(d 6 -DMSO) 8 8.39 (d, 1H), 5.72 (m, 1H), 5.15-5.2 (dd, 2H), 4.5 (m, 1H), 4.37 (m, 1H), 3.65 (m, 1H), 2.85-3.04 (m, 3H), 2.6-2.85 (m, 2H), 2.4 (m, 1H), 1.83-1.95 (m, 1H), 1.86 (s, 3H), 1.20 (t, J=7.46 Hz, 3H). MS: (M-H)- =301; (M+H) + = 303, (M+Na) = 325 -452- WO 99/54299 PCT/US99/07945 Example 216 (±)-(2R,3S,5R.1 'R, 3'R)-2-(1 -Acetamido-2-ethylsulfinyl)ethyl-3-vinvil-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt AcHN. OH S H O TFA The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,3'R)-1l-t-butoxycarbonyl-2-(1-N-t butoxycarbonylacetamido-2-ethylsulfinyl)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl 2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 12 mg, 94%). 1 H NMR(d 6 -DMSO) 8 8.39 (d, 1H), 5.72 (m, 1H), 5.15-5.2 (dd, 2H), 4.53 (m, 1H), 4.41 (t, 1H), 3.65 (m, 1H), 3.2 (dd, 1H), 2.9-3.0 (m, 2H), 2.65-2.9(m, 2H), 2.4(m, 1H), 1.83-1.95 (m, 1H), 1.83 (s, 3H), 1.20 (t, J=7.46 Hz, 3H) MS: (M-H)- =301; (M+H) + = 303, (M+Na) + = 325 -453- WO 99/54299 PCT/US99/07945 Example 217 (±)-(2R,3S,5R,1'R)-2-(1-N-t-butoxycarbonylacetamido-2-ethylsulfonyl)ethyl-3 vinyl-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt Ac BocN. OtBu HBoc6 o 217A (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-N-t-butoxycarbonylacetamido 2-ethylsulfonyl)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R,1'R,3'R)-1l-t-Butoxycarbonyl-2-(1-N-t butoxycarbonylacetamido-2-ethylsulfinyl)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (25 mg, 0.0448 mmole) was reacted with 55% m chloroperoxybenzoic acid (14 mg, 0.0448 mmole) in CHCI 3 (1.5 mL) at 0 oC for one hour. The reaction was concentrated in vacuo. The residue was purified by chromatography on silica gel using 25% ethyl acetate/hexane to provide the title compound (yield: 23.7 mg, 92%). 1 H NMR(d 6 -DMSO) 5 5.88 (m, 1H), 4.85-5.0 (m, 2H), 4.38 (m, 1H), 4.15 (m, 1H), 3.7 (m, 1H) 3.45 (dd, 1H), 2.9-3.2 (m, 3H), 2.5-2.7 (m, 1H), 2.3-2.4 (m, 3H), 1.6-2.04 (m, 1H), 1.35-1.55 (m, 27H), 1.15 (t, 3H) MS: (M+H) + = 575 -454- WO 99/54299 PCT/US99/07945 AcHN. OH C 0 TFA 217B (+)-(2R,3S,5R, 1'R)-2-(1-Acetamido-2-ethylsulfonyi)ethyl-3-vinyl-pyrrolidine 5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1-N-t butoxycarbonylacetamido-2-ethylsulfonyl)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 12 mg, 94%). 1 H NMR(d 6 -DMSO) 5 8.34 (d, 1H), 5.72 (m, 1H), 5.05-5.25 (dd, 2H), 4.68 (m, 1H), 4.39 (dd, 1H), 3.7 (2d, 1H), 3.48 (dd, 1H), 3.3-3.4 (dd, 2H), 3.08 (q, 2H), 2.95 (m, 1H), 2.42 (m, 1H), 1.9 (m, 1H), 1.84 (s, 3H), 1.23 (t, J=7.46 Hz, 3H). MS: (M-H) = 317, (M+35) + = 353; (M+H) + = 319, (M+Na) = 341 Examples 218, 220 OtBu 1.RSH AcHN. OH BocHN N 2. LiHMDS/AcCl N Oc 3. TFNCHC 2 s TFA The following title compounds were prepared in 3 steps according to the methods described in Example 214. -455- WO 99/54299 PCT/US99/07945 Example 218 (±)-(2R,3S,5R. 1'R)-2-(1-Acetamido-2-isopropylthio)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt BocH N. OtBu 'N fi 218A (±)-(2R,3S,5R, 1 'R)-1l-t-Butoxycarbonyl-2-(1-N-t-butoxycarbonylamino-2 isopropylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butvyl Ester. The title compound was prepared according to the method described in Example 214A, substituting isopropylthiol in place of ethanethiol (yield: 22 mg, 62%). 1 H NMR(d 6 -DMSO) 8 6.73 (d, 1H), 5.85 (m, 1H), 4.9-5.0 (m, 2H), 4.18 (m, 1H), 3.95 (m, 1H), 3.75 (br d, 1H), 2.8-3.0 (m, 2H), 1.65 (m, 1H), 1.32-1.45 (m, 27H), 1.18 (dd, 6H) MS: (M-H)- = 513; (M+H) + = 515, (M+Na) + = 537 Ac ' BocNK OtBu 218B (±)-(2R,3S.,5R, 1'R)-1l-t-Butoxycarbonyl-2-(1 -(N-t-butoxycarbonyvl-N acetamido)-2-isopropylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 214B, substituting (±)-(2R,3S,5R, 1 'R)-1l-t-Butoxycarbonyl-2-(1-N-t butoxycarbonylamino-2-isoproylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t -456- WO 99/54299 PCT/US99/07945 butyl ester in place of (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-N-t butoxycarbonylamino-2-ethylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 12 mg, 50%). 1 H NMR(d 6 -DMSO) 8 5.86 (m, 1H), 4.88-5.0 (m, 2H), 4.54 (m, 1H), 4.33 (m, 1H), 4.13 (d, 1H), 3.05 (m, 1H), 2.73-2.84 (m, 2H), 2.38 (brs, 3H), 1.72 (m, 1H), 1.32-1.5 (m, 27H), 1.14 (dd, 6H). MS: (M+H) + = 557, (M+Na) + = 579 AcHN, OH ~N .""I S H TFA 218C (±)-(2R.3S,5R. 1 'R)-2-(1-Acetamido-2-isopropylthio)ethyl-3-vinyl pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 15B, substituting (±)-(2R,3S,5R,1'R)-1l-t-butoxycarbonyl-2-(1-N-t butoxycarbonylacetamido-2-isopropylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 8 mg, 97%). 1 H NMR(d 6 -DMSO) 5 8.14 (d, 1H), 5.72 (m, 1H), 5.05-5.2 (dd, 2H), 4.2 4.4 (m, 2H), 3.68 (dd, 1H), 2.93 (m, 2H), 2.74 (dd, 1 H), 2.58 (dd, 1H), 1.93 (m, 1H), 1.87 (s, 3H), 1.2 (t, 6H) MS: (M-H)- = 299; (M+H) + = 301, (M+Na) + = 323 -457- WO 99/54299 PCT/US99/07945 Example 219 (±)-(2R,3S,5R,1'R)-2-( 1 -Acetamido-2-phenylthio)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Hydrochloride
H
2 N OtBu SHBoc a 219A (±)-(2R,3S,5R, 1 'R)-1l-t-Butoxycarbonyl-2-(1-amino-2-phenvlthio)ethyl-3 vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R, 3S,5R, 1 'S)-1 -t-Butoxycarbonyl-2-aziridinyl-3-vinyl-pyrrolidine-5 carboxylic acid t-butyl ester (20.3 mg, 0.06 mmole) was reacted with the phenylthiol (19.9 mg, 0.18 mmol) and triethylamine (0.047 mL, 0.34 mmol) in MeOH (0.06 mL) at ambient temperature for 3.5 hours. The reaction solution was concentrated in vacuo. The residue was purified by preparative thin layer chromatography on silica gel using ethyl acetate/methanol/ammonium hydroxide, 99/0.05/0.05, to provide the title compound (yield: 20.7 mg, 77%). 1H NMR (de-DMSO) 6 7.31 (m, 4H), 7.17 (m, 1H), 5.87 (m, 1H), 5.03 (d, J=17Hz, 0.4H), 5.01 (d, J=17Hz, 0.6H), 4.91 (d, J=11H, 0.4H), 4.90 (d, J=11Hz, 0.6H), 4.15 (m, 1H), 3.82 (m, 0.6H), 3.76 (m, 0.4H), 3.39(m, 1H), 2.92 (m, 2H), 2.55 (m, 1H), 1.64 (m, 2H), 1.42 (s, 5.4H), 1.37 (s, 3.6H), 1.34 (s, 5.4H), 1.22 (s, 3.6H) MS: (M+H) = 449, (M+Na) + = 471 -458- WO 99/54299 PCT/US99/07945 AcHN OtBu H Boc S 0 219B (±)-(2R, 3S,5R, 1'R)-1 -t-Butoxycarbonyl-2-(1-acetamido-2-phenylthio)ethyl 3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. (±)-(2R,3S,5R, 1'R)-1 -t-Butoxycarbonyl-2-(1 -amino -2-phenylthio)ethyl-3 vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (17.2 mg, 0.04 mmole) was reacted with the acetic anhydride (0.011 mL, 0.11 mmol) and triethylamine (0.032 mL, 0.23 mmol) in CH 2
CI
2 (0.3 mL) at rt for 4.25 hours. The reaction solution was concentrated in vacuo. The residue was purified by preparative thin layer chromatography on silica gel using 5% methanol/dichloromethane to provide the title compound. 1 H NMR (d 6 -DMSO) d 7.75 (d, J=9Hz, 0.6H), 7.73 (d, J=9Hz, 0.4H), 7.32 (m, 4H), 7.19 (m, 1H), 5.87 (m, 1H), 5.04 (d, J=17Hz, 0.4H), 5.00 (d, J=17Hz, 0.6H), 4.95 (d, J=10Hz, 0.6H), 4.93 (d, J=10Hz, 0.4H), 4.59 (m, 0.4H), 4.45(m, 0.6H), 3.99 (dd, J=10Hz, 2Hz, 0.6H), 3.94 (dd, J=10Hz, 2.5Hz. 0.4H), 3.84 (m, 0.6H), 3.77 (m, 0.4H), 3.07 (dd, 13Hz, 5Hz, 0.6H), 2.95 (m, 1.8H), 2.83 (brt, J=8Hz, 0.6H), 2.48 (m, 1H), 1.84 (s, 1.2H), 1.81 (s, 1.8H), 1.68 (m, 1H), 1.41 (s, 5.4H), 1.36 (s, 3.6H), 1.34 (s, 5.4H), 1.26 (s, 3.6H) MS: (M-H) = 489, (M+35); (M+H) = 490, (M+Na) = 513 -459- WO 99/54299 PCT/US99/07945 AcHN OH S H HCI 219C (±)-(2R,3S,5R,1'R)-2-(1-Acetamido-2-phenvithio)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Hydrochloride The title compound was prepared according to the method described in Example 1K, substituting (±)-(2R,3S,5R, 1 'R)-1 -t-butoxycarbonyl-2-(1-acetamido 2-phenylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±) (2R,3R,5R, 1 'S)-2-(1 -acetamido-3-ethyl)pentyl-3-(methoxymethyl)-pyrrolidine-5 carboxylic acid t-butyl ester (yield: 14.6 mg, 100%.) 1 H NMR(d 4 -methanol) 6 7.43 (m, 2H), 7.31 (m, 3H), 5.75 (ddd, J=17Hz, 10Hz, 8Hz, 1H), 5.32 (brd, J=17Hz, 1H), 5.19 (dd, J=10OHz, 1.4Hz, 1H), 4.58 (m, 2H), 3.89 (dd, J=10Hz, 3Hz, 1H), 3.19 (dd, J=14Hz, 6Hz, 1H), 3.09 (dd, J=14Hz, 9Hz, 1H), 3.04 (m, 1H), 2.57(dt, J=13Hz, 7Hz, 1H), 2.04 (s, 3H), 2.03 (m, 1H) MS: (M-H)- = 333; (M+H) + = 335, (M+Na) = 357 -460- WO 99/54299 PCT/US99/07945 Example 220 (±)-(2R,3S,5R,1'R)-2-(1-Acetamido-2-benzylthio)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt BocHN /OtBu S Boc 0 220A (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-N-t-butoxycarbonylamino-2 benzylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester The title compound was prepared according to the method described in Example 214A, substituting benzylmercaptan in place of ethanethiol (yield: 28 mg, 72%). 1 H NMR(d 6 -DMSO) 8 7.2-7.35 (m, 5H), 6.80 (br d, 1H), 5.84 (m, 1 H), 4.86-4.96 (m, 2H), 4.25(m, 1H), 3.95 (m, 1H), 3.7-3.8 (m, 3H), 2.76-2.94 (m, 1 H), 2.35-2.45 (m, 2H), 1.65 (m, 1H), 1.32-1.45 (m, 27H) MS: (M-H)- = 561; (M+H) + = 563, (M+Na) + = 585 -461- WO 99/54299 PCT/US99/07945 Ac BocN B.OtBu sooN 220B (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-(N-t-butoxycarbonyl acetamido)-2-benzylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 214B, substituting (±)-(2R,3S,5R, I'R)-1l-t-Butoxycarbonyl-2-(1-N-t butoxycarbonylamino-2-benzylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R)-1-t-Butoxycarbonyl-2-(1-N-t butoxycarbonylamino-2-ethylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 3.3 mg, 61%). 1 H NMR(d 6 -DMSO) 8 7.2-7.35 (m, 5H), 5.84 (m, 1H), 4.86-4.96 (m, 2H), 4.55(m, 1H), 4.32 (d, 1H), 4.05 (d, 1H), 3.56-3.65 (m, 2H), 2.9 (m, 1H), 2.3-2.65 (m, 3H), 2.42 (s, 3H), 1.76 (d, 1H), 1.25-1.55 (m, 27H) MS: (M+H) + = 605, (M+Na) + = 627 AcHN, OH P N "'l< S TFA 220C (±)-(2R,3S,5R,1'R)-2-(1-Acetamido-2-benzylthio)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1-N-t butoxycarbonylacetamido-2-benzylthio)ethyl-3-vinyl-pyrrolidine-5-carboxylic acid -462- WO 99/54299 PCT/US99/07945 t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 2.2 mg, 95%). 1 H NMR(d 6 -DMSO) 8 8.18 (d, 1H), 7.2-7.32 (m, 5H), 5.68(m, 1H), 5.02 5.2 (m, 2H), 4.3-4.45 (m, 2H), 3.76 (s, 2H), 3.68 (dd, 1H), 2.92 (m, 1H), 2.62 (dd, 1H), 2.32-2.55(m, 2H), 1.85-1.95 (m, 1H), 1.89 (s, 3H). MS: (M-H) = 347; (M+H) + = 349, (M+Na)* = 371 Example 221 AcHN, OH S S 2HCI (+)-(2R,3S,5R.1'R)-2-(1 -Acetamido-2-(4-pyridinethio)ethyl-3-vinyl-pyrrolidine-5 carboxylic Acid Dihydrochloride. The title compound was prepared according to the method of Example 219A-C substituting 4-thiopyridine for thiophenol as the reagent in Example 219A. 1 H NMR(d 4 -methanol) d 8.57 (d, J=7Hz, 2H), 7.97 (d, J=7Hz, 2H), 5.85 (ddd, J=17Hz, 10Hz, 9Hz, 1H), 5.40 (br d, J=17Hz, 1H), 5.25 (dd, J=17Hz, 10Hz, 1H), 4.67 (dt, J=10OHz, 4Hz, 1H), 4.47 (dd, J=10OHz, 8Hz, 1H), 4.01 (dd, J=10Hz, 4Hz, 1H), 3.68 (dd, J=14Hz, 5Hz, 1H), 3.45 (dd, J=14Hz, 10Hz, 1H), 3.16 (m, 1H), 2.65 (dt, J=14Hz, 7Hz, 1H), 2.07 (m, 1H), 2.04 (s, 3H) MS: (M-H)- = 334; (M+H)* = 336, (M+Na)* = 358 -463- WO 99/54299 PCT/US99/07945 Example 222
H
3 C AcHN,. .. OEt OH (±)-(2R,3S,5R.1'R,2'S)-2-(1-Acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Ethyl Ester. Thionyl chloride (1.49 mL, 20.5 mmol) was reacted with ethanol (25 mL) at 0 0 C for 10 minutes. (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy)butyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid trifluoroacetic acid salt (815 mg, 2.05 mmol) in ethanol (50 mL) was added to the above solution and reacted at room temperature for 17 hours. The reaction was concentrated in vacuo and the residue was purified by chromatography on silica gel with 90/10/0.5 dichloromethane / methanol / ammonium hydroxide to provide the title compound as a white solid (yield: 462 mg, 72%). 'H NMR (DMSO-d 6 ) 6 7.49 (d, J = 9.8 Hz, 1H), 5.31 (m, 2H) 4.11 (m, 2H), 3.72 (t, J = 7.7 Hz, 1H), 3.69 (m, 1H), 3.42 (m, 1H), 3.07 (m, 1H), 2.85 (m, 1H), 2.22 (m, 1H), 1.76 (s, 3H), 1.54 (d, J = 5.6 Hz, 3H), 1.45 (m, 1H), 1.39 (m, 1H), 1.21 (m, 1H), 1.19 (t, J = 7.0 Hz, 3H), 0.83 (t, J= 7.3 Hz, 3H). MS: (M+H) = 313, (M+Na) = 335, (M-H)- = 311. -464- WO 99/54299 PCT/US99/07945 Example 223 AcHN OEt OH O P OH (±)-(2R,3R,5R,1 'R,2'S)-2-(1 -Acetamido-2-hydroxy)butyl-3-(pyrazol-3-yI) pyrrolidine-5-carboxylic Acid Ethyl Ester The title compound is prepared according to the method described in Example 222, substituting (±)-(2R,3R,5R,1'R,2'S)-2-(1-acetamido-2 hydroxy)butyl-3-(pyrazol-3-yl)-pyrrolidine-5-carboxylic acid trifluoroacetic acid salt in place of (±)-(2R,3S,5R, I'R,2'S)-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic trifluoroacetic acid salt (yield: 32 mg, 52%). 1 H NMR(d 6 -DMSO) 68 7.6 (brs, 1H), 6.1 (brs, 1H), 4.08 (q, J=7.12Hz, 2H), 3.78 (m, 1H), 3.65 (m, 1H), 3.55 (m, 1H), 3.45 (m, 1H), 3.25 (m, 1H), 3.45 (m, 1H), 1.72 (s, 3H), 1.45 (m, 1H), 1.2 (m, 1H), 1.16 (t, J=7.12 Hz, 3H), 0.82 (t, J=7.46 Hz, 3H). MS: (M-H) = 337, (M+35) + = 373; (M+H) + = 339, (M+Na) = 361 -465- WO 99/54299 PCT/US99/07945 Example 224
H
3 C OEt AcHN. Qt H H 0 'CH 3 (±)-(2R,3S,5R,1'S,3'S)-2-(I -Acetamido-2-(N-isopropyl-N-methylamino-N oxide))ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid Ethyl Ester The title compound is prepared according to the method described in Example 222, substituting (±)-(2R,3S, 5R, 1'S,3'S)-2-(1-acetamido-2-(N-isopropyl) N-methylamino-N-oxide))ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid trifluoroacetic acid salt in place of (±)-(2R,3S,5R,1'R,2'S)-2-(1-acetamido-2 hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid trifluoroacetic acid salt (yield: 25 mg, 34%). 1H NMR (MeOD-d 3 ) 6 5.51-5.43(m, 1H), 5.34-5.27(m, 1H), 4.36-4.30(m, 1H), 4.18(q, J=7.1Hz, 2H), 3.88(dd, J=6.8, 8.8Hz, 1H), 3.82-3.67(m, 2H), 3.49 3.42(m, 1H), 3.34(s, 3H), 3.14-2.96(m, 1H), 2.42-2.33(m, 1H), 1.92(s, 3H), 1.64 1.52(m, 1H), 1.63(dd, J=1.7, 6.8Hz, 3H), 1.41-1.24(m, 1H), 1.39(d, J=6.4Hz, 3H), 1.31(d, J=6.4Hz, 3H), 1.26(t, J=7.1Hz, 3H). MS: (M+H)+=356, (M+Na)+=378, (M-H)-=354, (M+35)+=390. -466- WO 99/54299 PCT/US99/07945 Example 225
H
3 C AcHN. o, Et N HH O (±)-(2R,3S,5R. 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine 5-carboxylic Acid Ethyl Ester The title compound is prepared according to the method described in Example 222, substituting (±)-(2R,3S,5R, 1 'S)-2-(1-acetamido-3-methyl)butyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid trifluoroacetic acid salt in place of (±)-(2R,3S,5R,1'R,2'S)-2-(1 -acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic acid trifluoroacetic acid salt (yield: 838 mg, 94%). 1H NMR (CDCI 3 ):8 5.50 (m, 1H), 5.41 (m, 1H), 5.28 (m, 1H), 4.21 (q, J=7.5Hz, 2H), 4.06 (m, 1H), 3.87 (t, J=7.5Hz, 1H), 3.10 (m, 1H), 2.97 (m, 1H), 2.39 (m, 1H), 1.97 (s, 3H), 1.66 (dd, 3H), 1.60 (m, 1H), 1.40 (m, 2H), 0.94 (d, J=7.5Hz, 3H), 0.93 (d, J=7.5 Hz, 3H). MS: (M+H)+=311 -467- WO 99/54299 PCT/US99/07945 Example 226 Cl AcHN. OEt O N 'i H 0 (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(cis-2-chloro-vin-1-yl) pyrrolidine-5-carboxylic Acid Ethyl Ester The title compound is prepared according to the method described in Example 222, substituting (±)-(2R,3S,5R,1'S)-2-(1-acetamido-3-methyl)butyl-3 (cis-2-chloro-vin-1-yl)-pyrrolidine-5-carboxylic acid trifluoroacetic acid salt in place of (±)-(2R,3S,5R,1'R,2'S)-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1l-yl) pyrrolidine-5-carboxylic acid trifluoroacetic acid salt (yield: 28 mg, 46%). 1 H NMR (CDCl 3 ): 5 6.05 (d, J=7.5Hz, 1H), 5.90 (dd, J1=9 Hz, J2=6Hz, 1H), 5.31 (d, J=9Hz, 1H), 4.19 (q, J=7.5Hz, 2H), 4.06 (m, 1H), 3.82 (t, J=7.5Hz, 1H), 3.17 (m, 2H), 2.45 (m, 1H), 1.98 (s, 3H), 1.67 (m, 1H), 1.60 (m, 1H), 1.37 (m, 2H), 1.27 (t, J=7.5Hz, 3H), 0.91 (d, J=7.5Hz, 3H), 0.89 (d, J=7.5Hz, 3H). MS: (M+H) = 331 Example 227 Intentionally blank. -468- WO 99/54299 PCT/US99/07945 Example 228 F F AcHN. ., OEt 'N 7 HH0 (±)-(2R,3S,5R,1'S)-2-(1-Acetamido-3-methyl)butyl-3-(2,2-difluoro-vinyl) pyrrolidine-5-carboxylic Acid Ethyl Ester The title compound is prepared according to the method described in Example 222, substituting (±)-(2R,3S,5R,1'S)-2-(1-acetamido-3-methyl)butyl-3 (2,2-difluoro-vinyl)-pyrrolidine-5-carboxylic acid trifluoroacetic acid salt in place of (±)-(2R,3S,5R,1'R,2'S)-2-(1 -acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic acid trifluoroacetic acid salt (yield: 28 mg, 57%). 1 H NMR (CDCl 3 ): 8 4.22 (q, J=7.5 Hz, 2H), 4.14 (m, 1H), 4.03 (m, 1H), 3.29 (br, 1H), 2.85 (m, 1H), 2.52 (m, 1H), 2.01 (s, 3H), 1.77 (m, 2H), 1.64 (m, 2H), 1.49 (m, 1H), 1.38 (m, 1H), 1.29 (t, J=7.5Hz, 3H), 0.93 (d, J=7.5Hz, 3H), 0.90 (d, J=7.5Hz, 3H). MS: (M+H) = 333 -469- WO 99/54299 PCT/US99/07945 Example 229 H NN _, AcHN OEt 'N if H U (±)-(2R,3R,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(pyrazol-3-yl)-pyrrolidine-5 carboxylic Acid Ethyl Ester The title compound is prepared according to the method described in Example 222, substituting (±)-(2R,3R,5R,1'S)-2-(1-acetamido-3-methyl)butyl-3 (pyrazol-3-yl)-pyrrolidine-5-carboxylic acid trifluoroacetic acid salt in place of (±) (2R,3S,5R,1'R,2'S)-2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic acid trifluoroacetic acid salt (yield: 48 mg, 75.5%). 'H NMR (CDCI 3 ): 5 7.49 (d, 1H), 7.26 (s, 1H), 6.18 (d, 1H), 4.18 (q, J=7.5Hz, 2H), 4.12 (m, 1H), 3.91 (t, J=7.5Hz, 1H), 3.51 (t, J=7.5Hz, 1H), 3.40 (q, J=9Hz, 1H), 2.64 (m, 1H), 2.00 (m, 1H), 1.82 (s, 3H), 1.75 (m, 1H), 1.36 (m, 1H), 1.26 (t, J=9Hz, 3H), 0.855 (d, 3H), 0.84 (d, 3H). MS: (M+H)*= 337 -470- WO 99/54299 PCT/US99/07945 Example 230 (±)-(2R,3S,5R,1'R)-2-(1 -Acetamido-2-ethyl-2-hydroxy)butyl-3-(cis-propen-1-vi) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN. OtBu .N H Boc O OH 230A (±)-(2R,3S,5R,1'R)-1l-t-Butoxvcarbonyl-2-(1-acetamido-2-ethyl-2 hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butvl Ester. The title compound was prepared according to the method described in Example 41B, substituting (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1-acetamido 2-oxo)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester for (±) (2R,3S,5R,1'R)-l -t-butoxycarbonyl 2-(1 -acetamido-1l-formyl)methyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester to provide the title compound (yield: 0.021 g, 51%). MS: (M+H) + = 469, (M+Na) = 491, (2M+Na) =959, (M-H)- = 467.
CH
3 AcHN. OH ' N HH OH 0 TFA 230B (±)-(2R, 3S,5R, 1'R)-2-(1 -Acetamido-2-ethyl-2-hydroxy)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-ethyl-2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R, 1'R,2'S)-1 -t-butoxycarbonyl-2-(1 -471- WO 99/54299 PCT/US99/07945 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0039 g, 100%). 'H NMR (DMSO-d 6 ) 87.52 (d, J=10.3Hz, 1H), 5.45 (m, 1H), 5.28 (m, 1H), 4.32 (m, 2H), 3.68 (t, J=8.8Hz, 1H), 3.16 (quint., J=8.5Hz, 1H), 2.41 (dt, J=13.2,8.3Hz, 1H), 1.81(s, 3H), 1.59 (m, 1H), 1.53 (dd, J=6.8,1.5Hz, 3H), 1.52 1.42 (m, 3H), 1.30 (m, 1H), 0.86 (t, J=7.3Hz, 3H), 0.83 (t, J=7.3Hz, 3H). MS: (M+H) = 313, (M+Na) + = 335, (M-H) = 311, (2M-H)- = 623. Example 231 (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamido-2-hydroxy-2-methyl)pentyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN., N- H Boco OOH 231A (±)-(2R,3S.5R, 1 'R,2'S)-1 -t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy-2 methyl)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 41 B, substituting (±)-(2R,3S,5R, 1 'R)-1 -t-butoxycarbonyl-2-(1 -acetamido 2-oxo)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester for (-) (2R,3S,5R, 1'R)-1l-t-Butoxycarbonyl 2-(1-acetamido-1 -formyl)methyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester and methylmagnesium bromide for ethylmagnesium bromide to provide the title compound (yield: 0.0285 g, 45%). MS: (M+H) = 469, (M+Na) = 491. -472- WO 99/54299 PCT/US99/07945
CH
3 AcHN- OH H OH TFA 231B (±)-(2R.3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-2-methyl)pentyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy-2-methyl)pentyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0040 g, 100%). 1 H NMR (DMSO-d 6 ) 89.25 (bs, 1H), 8.75 (bs, 1H), 7.54 (d, J=10.3Hz, 1H), 5.45 (m, 1H), 5.29 (m, 1H), 4.37 (bt, J=8.3Hz, 1H), 4.22 (t, J=9.7Hz, 1H), 3.62 (t, J=8.8Hz, 1H), 3.12 (quint., J=8.5Hz, 1H), 2.41 (dt, J=12.7,7.8Hz, 1H), 1.78 (s, 3H), 1.59 (m, 1H), 1.53 (dd, J=6.8,2.0Hz, 3H), 1.4-1.25 (m, 4H), 1.17 (s, 3H), 0.81 (t, J=6.5 Hz, 3H). MS: (M+H) = 313, (M+Na)* = 335, (M-H)- = 311, (2M-H)- = 623 -473- WO 99/54299 PCT/US99/07945 Example 232 (±)-(2R,3S, 5R,1'R,2'S)-2-(1 -Acetamido-2-ethyl-2-hydroxy)pentyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN. O tBu N HBoc6 OH 232A (±)-(2R,3S,5R, 1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-ethyl-2 hydroxy)pentvyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 41B, substituting (±)-(2R,3S,5R,1'R)-1l-t-butoxycarbonyl-2-(1-acetamido 2-oxo)pentyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester for (±) (2R,3S,5R, 1 'R)-1 -t-butoxycarbonyl 2-(1 -acetamido-1l-formyl)methyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester to provide the title compound (yield: 0.0222 g, 33%). MS: (M+H) = 483, (M+Na)* = 505, (M-H)=481.
CH
3 AcHN. OH HH OH TFA 232B (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-ethyl-2-hydroxy)pentyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-ethyl-2-hydroxy)pentyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 -474- WO 99/54299 PCT/US99/07945 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0035 g, 100%). 1 H NMR (DMSO-de) 6 9.1 (bs, 1H), 8.75 (bs, 1H), 7.53 (d, J=9.8Hz, 1H), 5.44 (m, 1H), 5.28 (m, 1H), 4.35-4.25 (m, 2H), 3.67 (m, 1H), 3.16 (quint., J=8.5Hz, 1H), 2.41 (dt, J=12.8,7.9Hz, 1H), 1.81 (s, 3H), 1.60 (m, 1H), 1.53 (dd, J=6.7,1.8Hz, 3H), 1.46 (m, 2H), 1.4-1.20 (m, 4H), 0.86 (t, J=7.3Hz, 3H), 0.82 (t, J=6.7 Hz, 3H). MS: (M+H) = 327, (M-H)- = 325, (M+CF 3 COOH)-=439, (2M-H) = 651 Example 233
CH
3 AcHN. ' OH . N HH OH TFA (±)-(2R,3S,5R,1'R)-2-(1-Acetamido-2-propyvl-2-hydroxy)pentyl-3-(cis-propen-1-yvl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 232 substituting propyl magnesium bromide for ethyl magnesium bromide. 1 H NMR (DMSO-d6): 8 0.81 (t, 3H), 0.91 (t, 3H), 1.24-1.49 (m, 8H), 1.54 (dd, 3H), 1.60 (m, 1H), 1.81 (s, 3H), 2.41 (m, 1H), 3.15 (m, 1H), 3.69 (t, 1H), 4.28 (t, 1H), 4.35 (t, 1H), 5.17 (brs, 1H), 5.28 (td, 1H), 5.45 (dq, 1H), 7.54 (d, 1H), 8.80 (brs, 1H), 9.12 (brs, 1H). MS: (M+H)+= 341. -475- WO 99/54299 PCT/US99/07945 Example 234 (±)-(2R, 3S,5R, 1'R)-2-(1-Acetamido-2-ethyl-2-methoxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
CH
3 AcHN. OtBu N H Boc O ON~
S
234A (±)-(2R,3S,5R, 1'R)-1-t-Butoxycarbonyl-2-(1-Acetamido-2-ethyl-2 (methylthio)methyloxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic Acid t Butyl Ester (±)-(2R,3S,5R,1'R,2'S)-1 -t-Butoxycarbonyl-2-(1-acetamido-2-ethyl-2 methoxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester was reacted with dimethylsufoxide and acetic anhydride according to the method of Marshall, J. A. in J. Org. Chem. 1979, vol. 44, p 2994 to provide the title compound.
CH
3 AcHN. OtBu 'N ' H Boc O
OCH
3 234A (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-ethyl-2 methoxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butvl Ester (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-ethyl-2 (methylthio)methyloxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t -476- WO 99/54299 PCT/US99/07945 butyl ester is reacted with Raney Nickel according to the procedure of Marshall, J. A. in J. Org. Chem. 1979, vol. 44, p 2994 to provide the title compound. H3OH H H AN OH OCH 3 O TFA (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-ethyl-2-methoxy)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound is prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-ethyl-2-methoxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. Example 235 HOC Ac~ N. OH
OCH
3 STFA (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-ethyl-2-methoxy)pentyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound is prepared according to the method described in Example 234 substituting (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-ethyl-2-hydroxy)pentyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic -477- WO 99/54299 PCT/US99/07945 acid t-butyl ester for (±)-(2R,3S,5R, 1'R)-l -t-butoxycarbonyl-2-(1 -acetamido-2 ethyl-2-hydroxy)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester in 234A. Example 236 (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxymethyl-2-hydroxy)pentvl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
H
3 C AcHN. . OtBu oc Oq OH 236A (±)-(2R,3S,5R, 1 'R,2'S)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-((1 ethoxy)ethyloxvmethyl)-2-hydroxy)pentyl-3-(cis-propen- l -yl)-pyrrolidine-5 carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R, 1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-oxo)pentyl-3 (cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (50 mg, 0.11 mmole) was reacted with (ethoxyethyloxymethyl)tributylstannane (260 mg, 0.66 mmole) according to the method of Still, W. C. (J. Am. Chem. Soc., 100, 1481(1978)) to provide the title compound (yield: 26.8 mg, 43.8%). 1 H NMR (CDCI 3 ): 5 0.89 (t, 3H), 1.19 (m, 3H), 1.29 (dd, 3H), 1.45 (s, 9H), 1.46 (s, 9H), 1.52-1.73 (m, 8H), 1.99 (s, 3H), 2.44 (m, 1H), 3.24-3.74 (m, 5H), 3.91-4.22 (m, 3H), 4.49 (m, 1H), 4.62 (m, 1H), 5.37 (m, 1H), 5.64 (m, 1H), 5.97 6.41 (m, 1H). MS: (M+H)+= 557. -478- WO 99/54299 PCT/US99/07945
H
3 C AcHN, OH OH 0 OH TFA 236B (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxymethyl-2-hydroxy)pentyl 3-(cis-propen-1 -vyl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt (±)-(2R,3S,5R, 1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-(1-ethoxy-2 ethoxymethyl)-2-hydroxy)pentyl-3-(cis-propen- l -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester (13.5 mg, 0.024 mmol) was dissolved in THF (1 mL) and treated with 0.5 N HCI ( 1 mL) at room temperature for 1 hr. The solvents were removed and the resulting white solid was reacted with trifluoroacetic acid (0.8 mL) in dichloromethane (0.2 mL) at room temperature for 6 hours. The reaction was concentrated in vacuo overnight to provide the title compound (yield: 10.7 mg) as a off white solid. 'H NMR (DMSO-d 6 ): 6 0.81 (t, 3H), 1.24-1.38 (m, 4H), 1.52 (dd, 3H), 1.62 (m, 1H), 1.78 (s, 3H), 2.41 (m, 1H), 3.11 (m, 1H), 3.51 (qAB, 2H), 3.77 (t, 1H), 4.23 (t, 1H), 4.40 (m, 1H), 5.27 (t, 1H), 5.45 (m, 1H), 7.55 (d, 1H), 8.87 (br s, 1H), 9.26 (br s, 1H). MS: (M+H) = 329 -479- WO 99/54299 PCT/US99/07945 Example 237 (±)-(2R,3S, 5R,1'R,2'S)-2-(1 -Acetamido-2-allyloxy-2-vinyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt HC AcN. OtBu H Boc o 237A (±)-(2R,3S,5R, 1'R,2'S)-1-t-Butoxycarbonyl-2-(1-acetamido-2-allyloxy-2 vinyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester (2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy-2 vinyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester was reacted according to the method described in Example 84A substituting allyl iodide for methyl iodide (yield: 28 mg, 80%). MS: (M+H)
+
= 479, (M-H)- = 477 H3C AcHN OH 0_ 0 O) TFA 237B (±)-(2R,3S,5R.1'R,2'S)-2-(1-Acetamido-2-allyloxy-2-vinyl)ethyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-l -t-butoxycarbonyl-2-(1 acetamido-2-allyloxy-2-vinyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 -480- WO 99/54299 PCT/US99/07945 acetamido-2-hydroxy)butyl-3-(cis-propen- 1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester. (yield: 4 mg, 100%). 'H NMR (DMSO-d6) 6 7.98 (d, J=7.8 Hz, 1H), 5.90 (m, 1H), 5.55 (m, 1H), 5.48 (m, 1H), 5.32 (m, 2H), 5.26 (m, 2H), 5.16 (m, 1H), 4.28 (m, 2H), 3.96 (m, 1H), 3.79 (m, 1H), 3.73 (m, 1H), 3.66 (m, 1H), 3.26 (m, 1H), 2.40 (m, 1H), 1.81 (s, 3H), 1.70 (m, 1H), 1.64 (dd, J=6.9, 1.5 Hz, 3H). MS: (M+H) = 323, (M-H) = 321. Example 238 (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-1 -(2,5-dihydrofuran-2-yl))methyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
H
3 C AcHN. .,,OtBu 238A (±)-(2R. 3S,5R, 1'R.2'S)-2-(1 -Acetamido-1 -(2,5-dihydrofuran-2-yl))methyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-allyloxy-2 vinyl)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (21 mg, 0.044 mmole) prepared according to the procedure of Example 237A was reacted with bis(tricyclohexylphosphine)benzylidine ruthenium(IV) dichloride [Grubb's catalyst] (7.5 mg, 0.009 mmole) in methylene chloride (5 mL) at 25 0 C for 2 hours under a nitrogen atmosphere. The reaction was concentrated in vacuo and the resulting residue purified by chromatography on silica gel using 75% ethyl acetate/hexanes to provide the title compound (yield: 18 mg, 90%). -481- WO 99/54299 PCT/US99/07945 MS: (M+H)
+
= 451, (M-H) = 449. H3 AcHN. OH H 0 TFA 238B (±)-(2R,3S,5R, 1 'R,2'S)-2-(1-Acetamido-1l-(2,5-dihydrofuran-2-yl))methyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41C, substituting (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1 acetamido-1 -(2,5-dihydrofuran-2-yl))methyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester. (yield: 7 mg, 100%). 'H NMR (DMSO-d6) 6 8.09 (d, J=8.8 Hz, 1H), 6.10 (m, 1H), 5.87 (m, 1H), 5.50 (m, 1H), 5.27 (m, 1H), 4.68 (m, 2H), 4.58 (m, 1H), 4.33 (m, 1H), 4.06 (m, 1H), 3.68 (m, 1H), 3.18 (m, 1H), 2.40 (m, 1H), 1.85 (s, 3H), 1.68 (m, 1H), 1.60 (dd, J=6.8, 1.5 Hz, 3H), MS: (M+H) = 295, (M-H)- = 293. -482- WO 99/54299 PCT/US99/07945 Example 239 (±)-(2R, 3S, 5R, 1'R,2'S)-2-(1 -Acetamido-2-allyloxy-2-allyl)ethyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt H3C AcHN. OtBu Boc O 239A (±)-(2R,3S,5R,1 'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-allyloxy-2 alIlyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butvl Ester (2R,3S,5R, 1'R,2'S)-1 -t-Butoxycarbonyl-2-(1 -acetamido-2-hydroxy-2 allyl)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester was reacted according to the method described in Example 84A substituting allyl iodide for methyl iodide iodide (yield: 19 mg, 36%). MS: (M+H) = 493, (M-H) = 491. HC AcH-N. OH 'N H 0 0 TFA 239B (±)-(2R.,3S,5R,1'R,2'S)-2-(1-Acetamido-2-allyloxy-2-allyl)ethyl-3-(cis propen-1 -ylv)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 Acetamid o-2-al lyloxy-2-allyl)propyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 -483- WO 99/54299 PCT/US99/07945 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester. (yield: 5.7 mg, 100%). 1 H NMR (DMSO-d6) 8 8.06 (dd, J= 8.8 Hz, 1H), 6.92 (m, 1H), 6.77 (m, 1H), 5.50 (m, 1H), 5.29 (m, 2H), 5.17 (m, 1H), 5.05 (m, 2H), 4.27 (m, 2H), 4.10 (dd, J= 12.2, 5.4 Hz, 1H), 3.83 (m, 1H), 3.78 (m, 1H), 3.40 (m, 1H), 3.20 (m, 1H), 2.46 (m, 1H), 2.38 (m, 1H), 2.20 (m, 1H), 1.88 (s, 3H), 1.69 (m, 1H), 1.63 (dd, J= 6.8, 1.5 Hz, 3H). MS: (M+H) = 337, (M+Na) = 359, (M-H)- = 335. -484- WO 99/54299 PCT/US99/07945 Example 240 (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-1l-(3,6-dihydro-2-H-pyran-2-yl))methyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt
H
3 C AcHN, OtBu OH Boc Oi 0 240A (+)-(2R,3S,5R, 1'R.2'S)-2-(1-Acetamido-1 -(3,6-dihydro-2-H-pyran-2 yl))methy -3-(cis-propen-1 -yl)-pyrrolidine-5-carboxvlic Acid t-Butyl Ester (±)-(2R, 3S,5R, 1'R,2'S)-1-t-Butoxycarbonyl-2-(1-Acetamido-2-allyloxy-2-allyl)ethyl 3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (11.5 mg, 0.023 mmole) prepared according to the procedure of Example 239A was reacted with bis(tricyclohexylphosphine)benzylidine ruthenium(IV) dichloride [Grubb's catalyst] (3.8 mg, 0.005 mmole) in methylene chloride (3 mL) at 25 0 C for 3 hours under a nitrogen atmosphere. The reaction was concentrated in vacuo and the resulting residue purified by chromatography on silica gel using 75% ethyl acetate/hexanes to provide the title compound (yield: 5.7 mg, 53%). MS: (M+H) = 465, (M+Na)+= 487, (M-H) = 463 H C AcHN OH HN "~~ 0 0 0 TFA 240B (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-1l-(3,6-dihydro-2-H-pyran-2 yl))methyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt -485- WO 99/54299 PCT/US99/07945 The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-1-(3 ,6-dihydro-2-H-pyran-2-yl))pro pyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1l-t-butoxycarbonyl 2-(1-acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester. (yield: 5.9 mg, 100%). 1 H NMR (DMSO-d6) 6 8.04 (d, J= 8.8Hz, 1H), 5.77 (m, 2H), 5.50 (m, 1H), 5.25 (m, 1H), 4.21 (m, 2H), 4.14 (m, 1H), 4.04 (m, 1H), 3.81 (m, 1H), 3.40 (m, 1H), 3.23 (m, 1H), 2.41 (m, 1 H), 2.09 (m, 1H), 1.88 (s, 3H), 1.83 (m, 1H), 1.70 (m, 1 H), 1.63 (d, J= 6.8Hz, 3H). MS: (M+H) = 309, (M+Na) = 331, (M-H)- = 307 The following compounds were synthesized according to the methods previously described in Examples 1-240 Example 241 H3C AcHN OH ' O O HH 0 0 TFA (±)-(2R,3S,5R, 1'R,2'R)-2-(1 -Acetamido-1 -(3,6-dihydro-2-H-pyran-2-yl))provpyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1H NMR (DMSO-d6) 6 7.90 (d, 9.1Hz, 1H), 5.79 (m, 2H), 5.48 (m, 1H), 5.23 (m, 1H), 4.43 (m, 1H), 4.24 (m, 2H), 4.17 (m, 2H), 3.73 (m, 1H), 3.64 (m, 1H), 3.19 (m, 1H), 2.42 (m, 1H), 2.02 (m, 1H), 1.85 (s, 3H), 1.78 (m, 1H), 1.75 (m, 1H), 1.56 (dd, J= 7.5, 1.5 Hz, 3H). MS: (M+H) = 309, (M+Na) = 331, (M-H)- = 307. -486- WO 99/54299 PCT/US99/07945 Example 242 HC Ac N . OH OH 0 TFA (±)-(2R,3S,5R, 1'S,2'RS)-2-(1 -Acetamido-2-hydroxy)pentvl-3-(cis-propen-1 -vl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO) 8 7.7 (d, J= 9.8 Hz, 1H), 5.61 (m, 1H), 5.19 (dt, J= 1.8, 11.0 Hz, 1H), 4.33 (dd, J= 6.7, 10.3 Hz, 1H), 3.81 (m, 1H), 3.70 (dd, 1.8, 10.3 Hz, 1H), 3.54 (q, J= 6.1 Hz, 1H), 3.10 (m, 1H), 2.35 (dt, J= 12.8, 6.8 Hz, 1H), 1.90 (s, 3H), 1.7 (m, 1H), 1.59 (dd, J= 0.7, 7.3 Hz, 3H), 1.4 (m, 3H), 1.2 (m, 2H), 0.90 (t, J= 6.7 Hz, 3H). MS: (M+H) = 299 Example 243 HC AcN OH HN OH 0 0 OEt TFA (±)-(2R,3S,5R,1'S.2'RS)-2-(1-Acetamido-2-hydroxy-3-ethoxycarbonyl)propyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO) 6 7.75 (m, 1H), 5.60 (m, 1H), 5.29 (m, 1H), 4.55-4.25 (m, 3H),4.15-4.0 (m, 3H), 3.9-3.6 (m, 3H), 3.15 (m, 1H), 2.45-2.3 (m 2H), 1.9 (s, 3H), 1.8-1.5 (m, 5H), 1.2 (m, 3H). -487- WO 99/54299 PCT/US99/07945 MS: (M+H) = 343 Example 244
H
3 C AcHN OH 'N 'f H OCH3 O TFA (±)-(2R, 3S,5 R, 1'R,2'S)-2-(1 -Acetamido-2-methoxy-2-vinyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-de) d 7.91(d, J=8.05Hz, 1H), 5.50(m, 2H), 5.30(m, 3H), 4.27(m, 1H), 4.23(m, 1H), 3.75(m, 1H), 3.48(m, 1H), 3.23(m, 1H), 3.15(s, 3H), 2.40(m, 1H), 1.80(s, 3H), 1.68(m, 1H), 1.64(dd, J=1.83, 7.32Hz, 3H) MS: (M+H) 4 = 297, (M-H) = 295 Example 245
H
3 C AcHN. OH HN" TFA (±)-(2R,3S,5R.1'R,2'S)-2-(1-Acetamido-2-ethoMxy-2-vinyl)ethyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR(DMSO-ds) d 7.90(d, J=7.85Hz, 1H), 5.57(m, 2H), 5.48(m, 3H), 4.27(m, 1H), 4.22(m, 1H), 3.77(m, 1H), 3.60(m, 1H), 3.46(m, 1H), 3.23(m, 2H), 2.39(m, 1H), 1.80(s, 3H), 1.70(m, 1H), 1.64(dd, J=1.47, 6.73Hz, 3H), 1.12(t, J=6.83 Hz, 3H) -488- WO 99/54299 PCT/US99/07945 MS: (M+H) = 311, (M-H) = 309 Example 246
H
3 C AcHN. OH OH O TFA (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamido-2-hydroxy-2-(propeny-2-yl))ethyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR(DMSO-d 6 ) 5 7.69(d, J=9.75Hz, 1H), 5.47(m, 1H), 5.28(m, 1H), 5.03(m, 1H), 4.86(m, 1H), 4.40(m, 1H), 4.30(m,1H), 4.18(m, 1H), 3.97(m, 1H), 3.68(m, 1H), 3.21(m, 1H), 2.43(m, 1H), 1.82(m, 1H), 1.73(s, 3H), 1.64(s, 3H), 1.59(m, 3H) MS: (M+H) = 297, (M-H)-= 295 Example 247 HC Ac N. . OH 'NH 'OH 0 TFA (±)-(2R,3S,5R, 1 'R,2'R)-2-(1 -Acetamido-2-hydroxy-2-(propeny-2-yl)ethyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 6 7.65(d, J=9.80 HZ, 1H), 5.48(m, 1H), 5.23(m, 1H), 4.99(s, 1H), 4.88(s, 1H), 4.46(m, 1H), 4.30(m, 1H), 4.19(m, 1H), 3.55(m, 1H), -489- WO 99/54299 PCT/US99/07945 3.22(m, 1H), 2.44(m, 1H), 1.78(s, 3H), 1.75(m, 1H), 1.65(s, 3H), 1.58(dd, J=1.23, 6.70HZ, 3H) MS: (M+H)* = 297, (M-H)-=295 Example 248
H
3 C AcHN. OH N H0
OCH
3 TFA TFA (+)-(2R,3S,5R.1'R.2'S)-2-( 1 -Acetamido-2-methoxy-2-(propeny-2-yl))ethyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) d 7.77(d, J=9.8 Hz, 1H), 5.49 (m, 1H), 5.25(m ,1H), 5.07(m, 1H), 4.94(m, 1H), 4.32(m, 1H), 4.25(m, 1H), 3.75(m, 1), 3.48(m, 1H), 3.25(m, 1H), 3.08(s, 3H), 2.40(m, 1H), 1.77(s, 3H), 1.68(m, 1H), 1.64(dd, J=1.22, 6.71Hz, 3H), 1.56(s, 3H) MS: (M+H)+=311, (M-H)-=309 -490- WO 99/54299 PCT/US99/07945 Example 249 H C AcHN OH HH cN 0 TFA (±)-(2R,3S,5R. 1'R)-2-(1-Acetamido-2-ethyl)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 6 7.62(d, J=9.21Hz, 1H), 5.58(m, 1H), 5.28(m, 1H), 4.37(m, 1H), 3.98(m, 1H), 3.57(m, 1H), 3.10(m, 1H), 2.45(m, 1H), 1.92(s, 3H), 1.76(m, 1H), 1.62(dd, J=1.83, 6.72Hz, 3H), 1.24(m, 5H), 0.84(t, J=7.61Hz, 3H), 0.77(t, J=7.61Hz, 3H) MS: (M+H)+=297, (M-H)=295 Example 250
H
3 C AcHN. .. OH N HH O TFA (±)-(2R,3S,5R, 1 'S)-2-(1 -Acetamido-2-ethyl)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 5 7.76(d, J=9.2 Hz, 1H), 5.46(m, 1H), 5.29(m, 1H), 4.23(m, 1H), 3.63(m, 1H), 3.15(m, 1H), 3.01(m, 1H), 2.38(m, 1H), 1.87(s, 3H), 1.71(m, 1H), 1.60(m, 3H), 1.36(m, 1H), 1.20(m, 4H), 0.83 (t, J=7.3Hz, 6H) MS: (M+H)+=297, (M-H)-=295 -491- WO 99/54299 PCT/US99/07945 Example 251 HC HC AcHN OEt Ac 4 N OEt HH N HH HH O OH 'OH (-)-(2R,3S,5R,1 'S)-2-(1 -Acetamido-2-ethyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5 carboxylic Acid Ethyl Ester and (+)-(2S,3S,5S, 1'R)-2-(1 -Acetamido-2-ethyl)butyl 3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Ethyl Ester (±)-(2R,3S,5R, 1'S)-2-(1 -Acetamido-2-ethyl)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic acid ethyl ester (100 mg) was chromatographed in one injection on a chiral HPLC column of dimensions 5 x 30 cm. The column was packed with Chiralpak AD chiral stationary phase packing from Chiral Technologies. The mobile phase consisted of 1:9 ethanol:hexanes at a flow rate of 117 mL/min. Two peaks were observed at (24-36) minutes (-)-(2R,3S,5R,1'S) (yield: 45 mg) and at (66-96) min (+)-(2S,3S,5S,1'R) (yield: 45 mg). (-)-(2R,3S,5R,1'S) [aID = -260 (c=0.78, dichloromethane) -492- WO 99/54299 PCT/US99/07945 Example 252 (-)-(2R. 3S.5 R, 1 'S)-2-(1-Acetamido-2-ethyl)butyl-3-(cis-propen-1-yl)-pyrrolid ine-5 carboxvlate Ammonium Salt
H
3 C AcHN . . O NH OH 0 (-)-(2R,3S,5R,1'S)-2-(1-Acetamido-2-ethyl)butyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic acid ethyl ester (4.9 mg, 0.0157 mmole) prepared according to the procedure of Example 251 was reacted with lithium hydroxide (0.75 mg, 0.0314 mmole) in a mixture of methanol (0.75 mL) and water (0.25 mL) at 0 0 C for 7 hours. Then 0.1 N aqueous Hydrochloric acid (1 mL) was added, the reaction was concentrated in vacuo and the resulting residue purified by ion exchange chromatography on Aldrich Dowex 50WX8-400 strongly acidic resin. The residue was placed on the column and washed with water (5 mL) followed by elution using 0.5 N aqueous Ammonium hydroxide to provide the title compound as a colorless solid (yield: 3.9 mg, 83%). [a]D = - 400, c=0.08 (water). 1H NMR (DMSO-d6) 8 7.71 (d, J= 9.2 Hz, 1H), 5.38 (m, 1H), 5.29 (m, 1H), 3.92 (m, 1H), 3.65 (t, J= 8.5 Hz, 1H), 3.43 (m, 1H), 3.33 (m, 1H), 2.98 (m, 1H), 2.23 (m, 1H), 1.76 (s, 3H), 1.54 (dd, J= 6.7, 1.8 Hz, 3H), 1.46 (m, 2H), 1.23 (m, 1H), 0.84 (t, J= 7.3 Hz, 3H). MS: (M+H)= 285, (M+Na)+= 307, (M-H)- = 283. [aID = - 40, (c=0.08, water). Example 253 -493- WO 99/54299 PCT/US99/07945
H
3 C AcHN. OH ' N HH
SOCH
3 O
OCH
3 TFA (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2,3-dimethoxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (MeOD-d 3 ) 8.7.8(d, J=9.3Hz, 1H), 5.49-5.43(m, 1H), 5.25(dd, J=1.95, 9.3Hz, 1H), 4.38-4.31(m, 2H), 3.57-3.50(m, 1H), 3.46(dd, J=4.9, 10.3Hz, 1H), 3.42(s, 3H), 3.35-3.32(m, 2H), 3.27(s, 3H), 3.16-3.09 (m, 1H), 2.46-2.40(m, 1H), 1.80(s, 3H), 1.72-1.65(m, 1H), 1.55(d, J=6.8Hz, 3H). MS: (M+H)+=315, (M+Na) =337, (M-H)-=313, (M+Cl)=349, (2M-H)-=627. Example 254
H
3 C AcHN. N- OH N H H
OCH
3 O
OCH
3 TFA (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2,3-dimethoxy)propyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (MeOD-d 3 ) 5.8.04(d, J=8.5Hz, 1H), 5.52-5.48(m, 1H), 5.27 5.22(m, 1H), 4.32-4.25(m, 2H), 3.74-3.71(m, 1H), 3.53(dd, J=2.4, 10.1Hz, 1H), 3.33-3.25(m, 2H), 3.31(s, 3H), 3.25(s, 3H), 3.21-3.17(m, 1H), 2.42-2.36(m, 1H), 1.86(s, 3H), 1.71-1.63(m, 1H), 1.62(d, J=7.3Hz, 3H). MS: (M+H)+=315, (M+Na) =337, (M-H)-=313, (M+CI)-=349, (2M-H)-=627 -494- WO 99/54299 PCT/US99/07945 Example 255
H
3 C AcHN. . OH SN H H0 OH OH TFA (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxyethyl-2-hydroxy)pentyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ): 8 7.60 (m, 1H), 5.46 (m, 1H), 5.30 (m, 1H), 4.54 (m, 1H), 4.35 (m, 1H), 4.03 (m, 1H), 3.96 (m, 1H), 3.69 (m, 1H), 3.15 (m, 1H), 2.40 (m, 1H), 1.98 (m, 2H), 1.80 (s, 3H), 1.70-1.50 (m, 5H), 1.38 (m, 3H), 0.83 (m, 3H). MS: (M+H)+=343, (M-H)-=341 Example 256
H
3 C AcHN OH TFA (±)-(2R,3S,5R, 1 'R)-2-(1-Acetamido-2-(3-pentyloxy))ethyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (MeOD-d 3 ) 6 5.69-5.59 (m, 1H), 5.33-5.25 (m, 1H), 4.39 (m, 1H), 4.34 (dd, J=7.8, 10.2Hz, 1H), 3.73 (dd, J=4.8, 10.2Hz, 1H), 3.58-3.47 (m, 2H), -495- WO 99/54299 PCT/US99/07945 3.38-3.24 (m, 1H), 3.27-3.20 (m, 1H), 2.61-2.52 (m, 1H), 2.02 (s, 3H), 1.90-1.78 (m, 1H), 1.70 (dd, J=1.7, 6.8Hz, 3H), 1.60-1.50 (m, 4H), 0.92 (t, J=7.5Hz, 6H) (M+H) = 327, (M+Na)* = 349 Example 257 1=,,
H
3 C AcHN OH O _ H H O H H 0 TFA (+)-(2R, 3S,5R. 1 'S)-2-(1 -Acetamido-2-(3-pentyloxy))ethyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (MeOD-d 3 ) 5 5.73-5.66 (m, 1H), 5.32-5.25 (m, 1H), 4.36 (dd, J=7.8, 10.2Hz, 1H), 4.09 (m, 1H), 3.68 (dd, J=6.1, 10.2Hz, 1H), 3.61 (d, J=4.4Hz, 2H), 3.35-3.23 (m, 1H), 3.24-3.16 (m, 1H), 2.65-2.55 (m, 1H), 2.03 (s, 3H), 1.92 1.80 (m, 1H), 1.70 (dd, J=2.0, 7.1Hz, 3H), 1.59-1.47 (m, 4H), 0.94-0.88 (m, 6H) (M+H) = 327, (M+Na)+ = 349 -496- WO 99/54299 PCT/US99/07945 Example 258 H3C AcHN, OH ' N H0 0 TFA (±)-(2R, 3S, 5 R, 1'R,2'S)-2-(1 -Acetamido-2-ethoxy-3-vinyl)propyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 6 8.01 (d, J= 8.6Hz, 1H), 5.76 (m, 1H), 5.49 (m, 1H), 5.25 (m, 1H), 5.05 (m, 2H), 4.28 (m, 1H), 4.02 (m, 1H), 3.77 (m, 1H), 3.62 (m, 1H), 3.36 (m, 1H), 3.29 (m, 1H), 3.18 (m, 1H), 2.43 (m, 1H), 2.38 (m, 1H), 2.16 (m, 1H), 1.87 (s, 3H), 1.69 (m, 1H), 1.63 (dd, J=6.7, 1.2 Hz, 3H), 1.12 (t, J=6.7 Hz, 3H). MS: (M+H)= 325, (M-H)- = 323 Example 259 HC AcN. ,OH -H O 0 TFA (+)-(2 R, 3S,5 R. 1 'R)-2-(1 -Acetamid o-2-allyloxy)ethyl-3-(cis-p rope n- 1 -vyl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-d 6 ) 5 9.16 (m,2H), 8.13(d,J=7.5Hz,1H), 5.88(m,1H), 5.50 (m, 1H), 5.15-5.32(m, 3H), 4.35(m,2H), 3.95(m,2H), 3.61(m,1H), 3.40(m,2H), 3.20(m, 1H), 2.40(m,1H), 1.87(s,3H), 1.72(m,1H), 1.62(d, J=6.2,3H) -497- WO 99/54299 PCT/US99/07945 MS: (M+1)=297, (M+23)=319, (2M+23)=615 Example 260
H
3 C AcHN. OH N H OH 0 OEt TFA (t)-(2R,3S,5R,1'R,2'RS)-2-( -Acetamido-2-hydroxy-2-(2-ethoxycarbonvl))pentvl 3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (DMSO-de): 5 7.57 (d, J=10Hz, 1H), 5.45 (m, 1H), 5.29 (m, 1H), 4.35 (m, 1H), 4.09 (m, 1H), 3.68 (m, 1H), 3.44 (m, 1H), 3.17 (m, 1H), 2.87 (m, 1H), 2.64 (m, 1H), 2.39 (m, 1H), 1.80 (s, 3H), 1.65-1.56 (m, 2H), 1.53 (m, 3H), 1.50-1.30 (m, 3H), 1.21 (t, J=7.5Hz, 3H), 0.80 (t, J=7.5Hz, 3H). MS: (M+H)+=385, (M-H)-=383 Example 261 /=,,
H
3 C AcHN. H 'N HO.. HH 0 TFA HO -498- WO 99/54299 PCT/US99/07945 (±)-(2R,3S,5R, 1'S,3'R)-2-( 1 -Acetamido-3,4-dihydroxy)butyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (CD 3 OD) 8 5.58-5.70 (m, 1H), 5.24-5.38 (m, 1H), 4.34-4.50 (m, 2H), 3.58-3.72 (m, 2H), 3.42-3.48 (d, 2H), 2.50-2.63 (m, 1H), 2.04 (s, 3H), 1.77 1.95 (m, 1H), 1.65-1.76 (m, 4H), 1.50-1.63 (m, 1 H). MS: (M+H)*= 301 Example 262 /=,,
H
3 C AcHN. OH " N HO H 0 TFA HO (±)-(2R,3S,.5R.I'S,3'S)-2-(1-Acetamido-3,4-dihydroxy)butyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt 1 H NMR (CD 3 OD) 8 5.58-5.72 (m, 1H), 5.25-5.37 (m, 1H), 4.30-4.45 (m, 2H), 3.63-3.77 (m, 2H), 3.44-3.49 (d, 2H), 2.50-2.63 (m, 1H), 2.03 (s, 3H), 1.76 1.95 (m, 2H), 1.65-1.75 (m, 4H). MS: (M+H)* = 301 -499- WO 99/54299 PCT/US99/07945 Example 263 (±)-(2R, 3S, 5R, 1'R)-2-(1 -Acetamido-2-methoxy)ethyl-3-(cis-prope n-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN. OtBu H Boc
H
3 COO 263A (±)-(2R,3S,5R,1'R)-1l-t-Butoxycarbonyl-2-(1-Acetamido-2-methoxy)ethyl-3 (cis-propen-1-yi)-pyrrolidine-5-carboxvlic Acid t-Butyl Ester. The title compound was prepared according to the method described in Example 84A, substituting (±)-(2R,3S,5R,1'R)-1l-t-butoxycarbonyl-2-(1-acetamido 2-hydroxy)ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester for (±)-(2R,3S,5R, 1' R,2'S)-1-t-butoxycarbonyl-2-(1-acetamido-2-hydroxy)butyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 4.2 mg, 20%). MS: (M+H) =427, (M+Na)*=449, (M-H)-=425. r=,
CH
3 AcHN. OH
H
3 CO TFA 263B (±)-(2R,3S,5R, 1'R)-2-(1 -Acetamido-2-methoxy)ethyl-3-(cis-propen- 1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt. The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R)-1l-t-butoxycarbonyl-2-(1 acetamido-2-methoxy)ethyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1-acetamido -500- WO 99/54299 PCT/US99/07945 2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 0.0031 g, 100%). 1 H NMR (DMSO-d 6 ) 5 8.12 (d, J=7.9Hz, 1H), 5.50 (m, 1H), 5.23 (m, 1H), 4.33 (m 1H), 3.56 (dd, J=9.7,8.0Hz, 1H), 3.4-3.3 (m, 2H), 3.26 (s, 3H), 3.19 (m, 1H), 2.39 (dt, J=12.8,7.3Hz, 1H), 1.86 (s, 3H), 1.71 (m, 1H), 1.61 (dd, J=6.7,1.8Hz, 3H). MS: (M+H)*=271, (M+Na)*=293. Example 264 (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-dimethyl)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt.
CH
3 AcHN. OBu H Boc O OH 264A (±)-(2R,3S,5R,1'R,2'S)-1l-t-Butoxycarbonyl-2-(1-acetamido-2-hydroxy-3 dimethyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid t-Butyl Ester. The title compounds were prepared according to the method described in Example 41B, substituting t-butyl lithium for ethyl magnesium bromide to provide (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1-acetamido-2-hydroxy-3 dimethyl)butyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 2.5 mg, 11%) (±)-(2R,3S,5R,1'R,2'S) MS: (M+H) = 469; (M-H) = 467. -501- WO 99/54299 PCT/US99/07945
CH
3 AcHN. OH N HH OH TFA 264B (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy-4-vinvl)butyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt The title compound was prepared according to the method described in Example 41 C, substituting (±)-(2R,3S,5R, 1'R,2'S)-1l-t-butoxycarbonyl-2-(1 acetamido-2-hyd roxy-3-dimethyl)butyl-3-(cis-propen-1-yl)-pyrrol idine-5-carboxylic acid t-butyl ester in place of (±)-(2R,3S,5R,1'R,2'S)-1-t-butoxycarbonyl-2-(1 acetamido-2-hydroxy)butyl-3-(cis-propen-1-yl)-pyrrolidine-5-carboxylic acid t-butyl ester (yield: 2.3 mg, 100%). 1 H NMR (D 2 0) 5 5.40 (m, 1H), 5.10 (t, J=5.5Hz, 1H), 4.13 (t, J=9.2Hz, 1H), 3.46 (m, 1H), 3.22 (d, J=7.3Hz, 1H), 3.00 (m, 1H), 2.41 (m, 1H), 1.70 (s, 3H), 1.45 (m, 1H), 1.39 (d, J=4.9Hz, 3H), 1.07 (t, J=5.5Hz, 1H), 0.70 (s, 9H) MS: (M+H) = 313. Using the methods described above and the general knowledge of one skilled in the art, compounds of the invention can be prepared which are represented by taking one core from Table 1 (wherein Ac is acetyl), one Y substituent from Table 2, one R substituent from Table 3 and one R 3 substituent from Table 4a, 4b, 4c, 4d, 4e, 4f or 4g. -502- WO 99/54299 PCT/US99/07945 Table 1 Y, Y, AcHN OR AcHN OR R3 AH O R3 H O 1 2 Y, Y,
CH
3
SO
2 -HN OR CH 3
SO
2 +iN OR N 'NT(3P R3H l H O R H O 3 4 0 Y, 0 Y, II II
CF
3 C-HN OR CF 3 C-HN OR N fr'.'N R3 H O R3H H O 5 6 Table 2 H H 3 C H H H N
H
3 C ~ H - CI 1 2 3 4
H
3 C
H
3 C O
H
3 c 5 6 7 8 rN H S N F H F 9 10 11 12 -503- WO 99/54299 PCT/US99/07945 HN "N N O F 3 C\F HN-4 N 13 14 15 16
H
3 C H 3 C H F 3 C F -" Ci > F 3 C H 17 18 19 20
H
3 C F HN N N O N O __ N NZo
H
3 C 21 22 23 24
F
3 C F F 3 C N S 3F - FC - Cl 25 26 27 28 NH F H F H N4N Et - Et Et 29 30 31 32
F
3 C CI F CI Et H3C __O F - F 3 C - H -
'
3 33 34 35 36 H CH 3 F HN C H C C - H 37 38 39 40 HO 07 N N-N 41 42 43 44 -504- WO 99/54299 PCT/US99/07945 H N N N N N N HN N N N N-N 45 46 47 48 N, N , N N N HN' N HN 'N HN N I IN 49 50 51 52 H NN 0 - N-N 53 54 55 56 57 58 59 60 61 62 63 64 F H F F H - F F 65 66 67 68 F F F F CH 3
H
3 C CH 3 H F H3C 69 70 71 72
H
3 C CH CI C, C, CI C,
H
3 C - H CI 73 74 75 76 -505- WO99/54299 PCT/US99/07945 CI Cl CF 3
F
3 C CF 3 Cl CI 77 78 79 80 H C F 3 C Cl Cl CH 3 F3 C _ F 3 C -H 3 C - CI 81 82 83 84 Cl CH 3
F
3 C CF 3
H
3 C CI
H
3 C- CI - F 3 C -H 3 C 85 86 87 88 CI CH 3
H
3 C Cl H 3 C CH 3 CI CH 3 SCl , H3C 89 90 91 92 F F CH 3
H
3 C CI F - H 3 C - F - CI 93 94 95 96 Cl Cl CH 3 F F CH3
H
3 C/ - F/- H 3 C - H 3c 97 98 99 100
H
3 C F F CH 3
H
3 C F F F F -H 3 C 101 102 103 104
H
3 C F 3 C H 3 C CF 3
H
3 C CH 3
F
3 C - H 3 C - H 3 C - F 3 105 106 107 108 -506- WO 99/54299 PCT/US99/07945
H
3 C CF 3
F
3 C CH 3
CF
3
F
3 C CH 3
SH
3 C - F 3 C 109 110 111 112
CH
3
H
3 C CF 3
F
3 C CF 3 CI
F
3 C - F 3 C .- H 3 C ---- F 113 114 115 116 F C F Ci CI F CI F F 117 118 119 120 F Cl F CI Cl F F Cl 1- CI ~ F - Cl 121 122 123 124 CI F CI CI Cl Cl CF 3 F F 3 C F 3 C 125 126 127 128
F
3 C CI F 3 C C F 3 C C! F 3 C CF 3 F c C -~ c 129 130 131 132 CF CI Ci CF 3 CI CF 3 3 FCC Cl - F 3 C C0 - 3C 133 134 135 136 F CF 3 F CF 3 F CF3 F F - F - F 3 C ^ F 3 137 138 139 140 -507- WO 99/54299 PCT/US99/07945
F
3 C F F 3 C F CF 3
F
3 C F F - -F - F 3 C 141 142 143 144 F F 3 C CF 3
H
3 C F H 3 C CI
F
3 C '- F - CI F 145 146 147 148 CI F F Cl F CH 3 Cl CH 3
H
3 C - H 3 C CI F 149 150 151 152
H
3 C CF 3
H
3 C CI Cl 3
F
3 C>CI Ci - F 3 C - H 3 C - H 3 C 153 154 155 156
F
3 C CH 3 CI CH 3
H
3 C CF 3
H
3 C F Ci - F 3 C - F - F 3 157 158 159 160 F CF 3 F CF 3
F
3 C F F 3 C CH 3
H
3 C - H 3 C - H 3 C - F 161 162 163 164 F CH 3 CI CF 3 CI F F CF 3
F
3 C - F - F 3 C - CI 165 166 167 168 NH H 2 N H 2 N H2N \H 2~.. H 3 C 169 170 171 172 -508- WO 99/54299 PCT/US99/07945
H
2 N, NH NH 173 174 Table 3
-H
1
CH
3 2
C
3 3
CH
3 CH3 CH 3
CH
3 44 6
CH
3 5 4HCH3 CH3
JCH
3 )- CH 3 8O
H
3 C CH7 CH 3 H
C
3 \ CH 3 , CH 3 10 CH 3
H
3 12
\"CH
3
ICH
3
/-CH
3 - CH 3 13
\CH
3 14
\-CH
3 15 -509- WO 99/54299 PCT/US99/07945 Table 4a CH3 H 3 C OH
H
3 C OH OH H3OH
CH
3 1 2 3 4
CH
3 H 3 C OH OH H 3 C OH OH
CH
3
H
3 C o CH 3 H3
CH
3 56 78 O0HH OH 3H OH
H
3 C cHH CH 3 9 10 11 12
H
3 C H C HOH H H3OH OH H 3 30 13 14 15 16 S CH 3
CH
3 CH OCH 3
H
3 C -" OCH 3
OCH
3
H
3 C OCH 3 CH3 17 18 19 20
CH
3
-
H
3 C CH OCH H OCH 3
H
3 C OCH 3
H
3 C CH OH3 OH3 H3 C OH3H 21 22 23 24 -510- WO 99/54299 PCT/US99/07945
CH
3 -'' OCH OCH 3 3CH 3
OCH
3
HCOH
3 C~ COH 3 25 26 27 28
H
3 C
H
3 C CH OCCH H3OH3
CH
3
H
3 C CHOCH3
OCH
3 3 H 3 C CH 3 29 30 31 32 OH OH CH ,OH H 3 C X. H 3 C-OC
H
3 CH3H H 3 C 3OCH3 H 3 C COH H 3 C CH 3 33 34 35 36 H3 CJ H 3 C H
H
3 C ,OH H3 OH H 3 C OH HC 37 38 39 40 OH OH OH OH
H
3 C CH 3 41 42 43 44
H
3 C HOCC
H
3 C OCH 3 H3C
OCH
3
H
3 C
OCH
3 HzC
OCH
3 45 46 47 48 S-OCH, OC.
OCH
3 OCH 3 OCH 3 OCH 3
H
3 C OH 3 49 50 51 52 -511- WO 99/54299 PCT/US99/07945 Table 4b OH OCH 3 O CH3 OH OH OH OH CH 3 1 2 3 4 'OH 'OCH 3 O1- CH 3 o OH OH OH OH CH 3 5 6 7 8 HO OH HO OCHz HO 0 CH 3 HO O OH OH OH CH 3 9 10 11 12 HO OH HO OCH 3 HO O"CH 3 HO O OH 6OH H OH CH 3 13 14 15 16 HO 'OH HO 'OCH 3 HO 'OCH 3 HO 'O OH OH OH OH CH 3 17 18 19 20 HO ' OH HO 'OCH 3 HO 'O^CH 3 HO OH OH OH CHz 21 22 23 24 O 'O HO O HO 0 OH OH OH OH 25 26 27 28 -512- WO 99/54299 PCT/US99/07945 HO 'O HO 'O OCH 3
'OCH
3 OH OH OCH 3
OCH
3 29 30 31 32
CH
3 0 OCH 3
H
3 0 OCH 3 CH30 - OCH 3
CH
3 0 'OCH 3
OCH
3
OCH
3
OCH
3
OCH
3 33 34 35 36
H
3 C OoH H 3 C OCH HC O CH 3 H 3 C O OH OH OH OH CH 3 37 38 39 40 CH3.
H
3 C OH H 3 C OCH 3
H
3 C O CH 3
H
3 C O OH
H
3 C OH H 3C OH H 3 C OH CH 3 41 42 43 44
H
3 C Y O-H H 3 C Y OCH 3
H
3 C O- CH 3
H
3 C- o OH OH OH OH CH 3 45 46 47 48
H
3 Y OH H 3 C OCH 3
H
3 C---' O CH 3
H
3 C o OH OH OH OH CH 3 49 50 51 52
H
3 C -- OH H 3 C OCH 3
H
3 C-r CH HC \ o, OH OH OH OH CH 3 53 54 55 56
H
3 C OH H 3 C OCH 3
H
3 C , O'CH3 H 3 C OH OH OH OH CH 3 57 58 59 60 -513- WO 99/54299 PCT/US99/07945
H
3 C H 3 0 H 3 C 0 ___ C H 3 C ' 3 C 'OH H 3 C- OCH 3
H
3 C O H 3
H
3 C 0O OH OH OH OH CH 3 61 62 63 64 - - CH3,
H
3 0 'OH H 3 C OCH 3
H
3 C OCH 3
H
3 O OH H3COH
H
3 C OH H3 C OH H 3 OH OH 3 65 66 67 68
H
3 C OH H 3 , OH H 3 C OCH 3
H
3 C OCH 3 H3OH OH H --OH 3 OH OH OH OH 69 70 71 72
H
3 C -OH H 3 C ,OH H 3 C OCH 3
H
3 C OCH 3 H CH 3 6H -CH3 6H CH 3 6H '--CH 3 OH OH OH OH 73 74 75 76
H
3 C OH H 3 C OH H 3 C OCH 3
H
3 C OCH 3
H
3 C OH , H 3 0 '-OHC H 3 C OH 3 C H 3 C O CH 3 OH OH OH OH 77 78 79 80
-
CHy-- CH- H3 HC OH OH 3H 3
H
3 C OHH 3 H 3 '-OH 3 0 O-H 3
L-H
3 HC OH OH OH OH CH 3 81 82 83 84 Table 4c
CH
3
-
H
3 C HC -CH3 H 3 C - H 3
-CH
3 , CH OH OH OH H 3 CH3 2 3 4 -514- WO 99/54299 PCT/US99/07945
-OH
3
CH
3
CH
3
'OH
3
OH
3 3
HCH
3
CH
3 5 6 7 8
CH
3 - CH 3 CH3 H3C CH 3
'CH
3
H
3 C Z CH 3
CH
3
CH
3 9 10 11 12
CH
3 -~ OCH 3
'OH
3
H
3
CH
3
H
3 C COH 3
OH
3
H
3 C
OH
3
H
3 C OH 3 13 14 15 16
H
3
'CH
3
H
3 ' CH 3
'CH
3
CH
3
OH
3
H
3
OH
3 17 18 19 20
OH
3 H OH 3
OH
3 3 I'H H CH CHCH
'CH
3
CH
3 H3 3
H
3 C CH 3
OH
3
OH
3
H
3 C OH 3 21 22 23 24
OH
3
H
3 0
H
3
OH
3
OH
3 3CH 3
CH
3 OH OH OH OH 25 26 27 28
OH
3
OH
3
H
3 C S CH3 C OCH 3 3OH 3
OH
3 OH OH OH OH 29 30 31 32 -515- WO 99/54299 PCT/US99/07945 OH OH^ OH^ CH 'CH 3
CH
3
'CH
3 OH OH H 3 0 CH 3 33 34 35 36 CH3 -H 3 0 HCH3
H
3 C OH 3 O3,- H 3
OH
3 O-: H 3 37 38 39 40
CH
3 H HCHH 3
CH
3 3
CH
3 41 42 43 44
CH
3
CH
3 CH3 H 3 0 CH 3
CH
3
H
3 0 CH 3
OH
3
OH
3 45 46 47 48
CH
3 - CH 3
CH
3 3 0 CH 3 C CH 3 H H H 3
O
3 49 50 51 52
H
3 C- CH 3
H
3
C"CH
3
CH
3
CH
3 CH 3
OH
3 O3C CH 3 53 54 55 56 CH CH CH
OH
3 __ O.H 3 O ?H 3 / . ~H
CH
3
CH
3
CH
3
CH
3
H
3 C OH 3
OH
3
OH
3
H
3 C CH 3 57 58 59 60 -516- WO 99/54299 PCT/US99/07945 O H 3 H 3 C
H
3 O
CH
3 CH 3 OH 3 CH 3 OH OH OH OH 61 62 63 64
OH
3 OH H 3 C C CH 3
H
3 3CH 3
],CH
3 OH OH OH OH 65 66 67 68 OH^ OH OH '
CH
3
CH
3
CH
3 O. H 3 OH OH H 3 C OH 3 69 70 71 72
CH
3
H
3 0
H
3 C ,CH3 H3C. CH 3
.H
3
CH
3 73 74 75 76 " .L G3la .CH 3
CH
3
H
3 3
H
3
CH
3 77 78 79 80
CH
3
CH
3
OCH
3
H
3 0 ,- ClH 3
-H
3
H
3 O -CH 3
OH
3
CH
3 CH3 81 82 83 84
CH
3 - CH 3 _ .CH 3 H3 C l
CH
3
H
3C I CH 3
OH
3
H
3 C CH 3
H
3 C CH 3 85 86 87 88 -517- WO 99/54299 PCT/US99/07945
H
3 C 3 3 3
CH
3
CH
3
H
3 C OH 3 89 90 91 92
CH
3
H
3 CH 3 CH3 (;-, .- CH3 . H y -OH 3 .C3
H
3 C CH 3
OH
3
CH
3
H
3 C CH 3 93 94 95 96
C
OH
3
H
3 C
H
3
-CH
3
CH
3 j CH 3
CH
3 OH OH OH OH 97 98 99 100 C CH3 CH 3
H
3 C 17j _OH 3 , -H 3 yOH 3 .- ICH 3 OH OH OH OH 101 102 103 104 OH ~ OH OH CO CH r CH 3 O -- H 3 " ' C
H
3 OH OH HC CH 3 105 106 107 108
CH
3
H
3 C
H
3 C. CH 3
H
3 C ,.-CH 3
CH
3 l- CH 3 109 110 111 112 [ /CH3C1-!OH 3 113 114 115 116 -518- WO 99/54299 PCT/US99/07945 Qcwl
H
3 0C H3 3 O-- H 3 C CB C CH3 CC cI 117 118 119 120
-CH
3 , CH 3
CH
3
H
3 C CH 3
H
3 C CH 3
OH
3
H
3 C
CH
3 3 C OH 3 121 122 123 124
H
3 C CH 3
H
3 C CH 3
CH
3
CH
3
H
3 C0
OH
3
CH
3 125 126 127 128
OH
3
OH
3
CH
3 C CH H 3 CH 3 CH 3 CH 3
H
3 C CH 3
OH
3
OH
3
H
3 C CH 3 129 130 131 132
CH
3
H
3 C
H
3 C CH 3 CO H 3
CH
3
CH
3 OH OH OH OH 133 134 135 136
CH
3
H
3 C
H
3 Co COH 3
CH
3
HCH
3
CH
3 6H OH OH OH 137 138 139 140 OH OH - OH
HCO
3
H
3
CH
3
CH
3 OH H3C 0 0H3 141 142 143 144 -519- WO 99/54299 PCT/US99/07945
CH
3
CH
3
CH
3 - CH 3
H
3 C
H
3 3 H 3 C CH3 145 146 147 148
OH
3
CH
3
OH
3
OH
3
OH
3 149 150 150A 150B
CH
3
CH
3
CH
3
CH
3 - CH 3
CH
3 CH 3 C3 _H3 Y
H
3 C CH 3
CH
3 151 152 153 154
CH
3
CH
3
CH
3
CH
3 - H 3 ~ H 3
H
3 C
H
3 CH3
H
3 C
OH
3
H
3 0 CH 3 155 156 157 158
OH
3
CH
3
H
3
CH
3
H
3 C H 3 C3
H
3 C I C H
CH
3
CH
3 159 160 161 162
CH
3
CH
3
CH
3
CH
3
OH
3
H
3
OH
3
H
3 C CH 3
OH
3
OH
3
H
3 C OH 3 163 164 165 166 H3C H 3
CH
3
H
3 C CH 3 " H 3
H
3 C OH OH OH OH 167 168 169 170 -520- WO 99/54299 PCT/US99/07945
CH
3
CH
3
OH
3 3
H
3
OH
3 OH 6H OH 6H 171 172 173 174 CH OH
CH
3 OH
CH
3 OH
CH
3 CHi H3C OH OH H 3
CH
3 175 176 177 178
HH
3 HOC
H
3
CH
3
CH
3
H
3 C
H
3
H
3 C HC C 179 180 181 182
CH
3
OH
3
OH
3
OH
3
CH
3 183 184 185 186
CH
3 - CH 3
OH
3
CH
3
OH
3
OH
3
H
3 C 3 H 3 C H CCH3H 3
OH
3 187 188 189 190
CH
3
CH
3
CH
3
CH
3 - CH 3
H
3
H
3 C3 H 3 C H OH 3
H
3 C CH3 191 192 193 194
CH
3
CH
3
OH
3
CH
3
H
3 C H 3 C
CH
3
H
3 C OH 3 195 196 197 198 -521- WO 99/54299 PCT/US99/07945 CF- CH 3 H CH 3 O OH 3 W H 3
H
3 C CH 3
CH
3
CH
3 H3C CH 3 199 200 201 202
CH
3 CH OH 3
H
3 C OH 3 \ OH 3
H
3 Cw i OH OH OH OH 203 204 205 206 S
OH
3 O 3
H
3 O OH 3 \ OH 3 6H OH OH OH 207 208 209 210
OH
3 O H 3 OH OH 3 OH O H 3 OH OH H 3 C H 3 211 212 213 214
CH
3
H
3 C
H
3 0
H
3 , 215 216 217 218
CH
3 219 220 221 222
CH
3 -"" OH 3 - H3C .J, - H 3 ON
H
3 ON H
CH
3
CH
3 223 224 225 226 -522- WO 99/54299 PCT/US99/07945 ' CH3- CH 3
H
3 C
H
3 0
H
3 C
CH
3
H
3 0 OH 3 227 228 229 230
H
3 J H 3 C
CH
3 O3 H 3 231 232 233 234 CH CH CH 3
H
3 C CH 3
CH
3
CH
3
H
3 C CH 3 235 236 237 238 CH H 3 C H3C OH OH OH OH 239 240 241 242 CH C u H 3 C OH 6H OH OH 243 244 245 246 OH OH OH OH OH COH 3 247 248 249 250 3 3 C
-
H
3 0
H
3 O1.K) C 3 0 I 251 252 253 254 -523- WO 99/54299 PCT/US99/07945
CH
3 255 256 257 258
CH
3 CH3
H
3 C H 3 C r
CH
3
CH
3 259 260 261 262
CH
3
CH
3 33 H 3 C H 3 C HT"3 H 3 C
CH
3
H
3 C OH 3 263 264 265 266
H
3 C
H
3 C H3C 3 C
CH
3
CH
3 267 268 269 270 CH CHCH
H
3 C CH 3
CH
3 6H 3
H
3 c
CH
3 271 272 273 274
H
3 C CH
H
3 C OH OH OH OH 275 276 277 278
CH
3
CH
3 H3C OH OH OH OH 279 280 281 282 -524- WO 99/54299 PCT/US99/07945 OH OH OH 6H OH H 3 C OH 3 283 284 285 286 H CH 3 H H 3 C H3C 287 288 289 290
H
3 C
H
3 C
CH
3 H 3 C 291 292 Table 4d 1 2 3 4
H
3 C CH 3
H
3 C CH 3
H
3 C CH 3 5 6 7 8 O HC O HC' OH O HCH 3 C- ~ ,H 3 C S H 3 C_ 0 9 10 11 12 -525- WO 99/54299 PCT/US99/07945
H
3 C
H
3 C
H
3 C O
H
3 0 0 0 0 13 14 15 16 H3 0H 3 C- -0
H
3 C- -0 C H3 O H 3 O 17 18 19 20
H
3 C H 3 C H 3 O
H
3 C "0 21 22 23 24 0 0 6"0 0 25 26 27 28
H
3 C H 3 C I -0 H 3 C 6 0 H 3 C 0o 29 30 31 32
H
3 C H 3 C H 3 C
H
3 C _
H
3 C_
H
3 C_ 33 34 35 -526- WO 99/54299 PCTIUS99/07945 Table 4e T CH 3 - ~ H 3 C
H
3 C -1-OH H 3 C , OH -,OH 1 2 3 4 -- OH -N OH 0 H ~ OH
CH
3 5 6 7 8
CH
3 -- CH 3 ~OH H 3 C- , - IOH y ~ - -OH H 3 C-- N IOH
CH
3
CH
3 9 10 11 12
CH
3
CH
3 H3i OHIO H 3 C Ny */OH O
H
3 C
CH
3
H
3
CCH
3 13 14 15 16
H
3 C --OH H 3 C 11OH _/H _,OH
CH
3
HCCH
3 17 18 19 20
CH
3
CH
3
CH
3 ( O-1H (: ,,, OH [7 ,OH Y ,OH
H
3 C CH 3
CH
3 6H, H 3 C CH 3 21 22 23 24 -527- WO 99/54299 PCTIUS99/07945 OH
H
3 C
H
3 C JOH C OH OH -- OH OH OH OH OH 25 26 27 28
CH
3
"T"CH
3
H~
3 CO .OH C OH HOH OH OH OH OH OH 29 30 31 32 OH O OH OH O
..
OHH .OH OH u .. OH OH OH H 3 C H 3 33 34 35 36
CH
3 - H 3 C
H
3 OH H 3 C OH .H OH 37 38 39 40 SOH OH OH y OH
OH
3 41 42 43 44
CH
3
CH
3 : OH H 3 C OH OH H 3 C -- OH
CH
3
CH
3 45 46 47 48
CH
3
CH
3 -1 OH H 3 C OH H 3 C OH OH
H
3 C OH 3
H
3 C OH 3 49 50 51 52 -528- WO 99/54299 PCT/US99/07945
H
3 C OH H 3 C OH OH OH
OH
3
H
3 C OH 3 53 54 55 56
CH
3
"CH
3 CH C OH C OH OH O H
H
3 C CH 3
CH
3
OH
3
H
3 C CH 3 57 58 59 60 CH H 3 C
H
3 c OH OH OH OH OH OH OH OH 61 62 63 64
CH
3
H
3 C
H
3 C OH C OH H OH OH OH OH OH OH 65 66 67 68 OH ~ OH ~ OH OH O OH O H OH 0..5OH L...OO HC. O OH 3C H 3 69 70 71 72 OH OH HO OH oH 3 CO OCF 69 70 71 72 -529- WO 99/54299 PCT/US99/07945 Table 4f
CH
3
-
H
3 C
H
3 -OH3 H 3 C -CH 3
-CH
3 3 OH OH OH OH 3 CH3 1 2 3 4
-CH
3
OH
3
CH
3
H
3
OH
3
OH
3
OH
3 5 6 7 8
OH
3 - CH 3 c
HOC-OH
3
H
3 C CH 3 C H 3 3 OH 3 3 H 3 3 9 10 11 12
OH
3
CH
3 _
OH
3
H
3 C C 3
OH
3 ,OH 3 3 3
-,-OH
3
.OH
3 CH3
H
3 HC3
OCH
3
H
3
H
3
OCH
3 3 13 14 15 16
H
3 0 -OH 3
H
3 0 -OH 3
OH
3
OH
3 H OH 3
H
3 0 OH 3
H
3
H
3
OH
3
OH
3 " C 17 18 19 20
CH
3
CH
3
OH
3
H
3
-CH
3
-H
3 -CH3 3
CH
3
CH
3
OH
3
OH
3
H
3 C CH 3
OH
3
OH
3 H3C CH 3 21 22 23 24 -530- WO 99/54299 PCT/US99/07945
CH
3
H
3 C
H
3 C CH 3
CH
3
-OH
3
-OH
3
|OH
3 Z H 3 V OH 3 OH OH OH OH 3 OH 25 26 27 28
CH
3
OH
3
H
3 C CH
CH
3
CH
3 3 CCH 3
CCH
3 OH OH OH H 3 OH 29 30 31 32 OH OH OH
H
3
OCH
3
CH
3
-CH
3
OH
3 OH c H 3 / OH 3 OH OH
H
3 C OH 3 33 34 35 36
CO
3
H
3
O
3
H
3
H
3 C CH 3
H
3 C CH3 3
CH
3
OH
3
H
3 0 OH 3
CH
3 37 38 39 40
H
3 OH OH 3
H
3
CH
3
OH
3
CH
3
CH
3 41 42 43 44 CH3 CH 3 H H3 CH 3
CH
3
OH
3
H
3 C ~ CH H 3 0 c -~ CHc - H,C .
OH
3
OH
3
OH
3 OH 3
OH
3 CH3 3 45 46 47 48
CH
3
CH
3
H
3
O
3 C CH3 3 H 3 H 3 3
H
3 C HOH 3 H CH 3
CH
3
CH
3
H
3
H
3
OH
3 C H3C 3 H 49 50 51 52 -531- WO 99/54299 PCT/US99/07945
H
3 C 3 H 3
CH
3
CH
3
CH
3
CH
3
H
3 C OH 3
CH
3 O H 3
CH
3
CH
3 53 54 55 56
CH
3
OH
3 CH CH 3
CH
3
CH
3 OH 3
CH
3 O H 3
OH
3
H
3 C CH 3
CH
3
CH
3
H
3 C CH 3 57 58 59 60
SCH
3 CH3 3 H3C C C3
H
3 C -J
OH
3 3
OH
3
OH
3
H
3
CH
3 OH OH OH OH 61 62 63 64 CH OCH 3 CH H 3 CCH3 3y - Ha -T JCH 3 - CH 3 OH H OH H 3 OH 3 65 66 67 68 CH3 O C OH CH3 O CH 3 OH -- CH 3 O
OH
3 CH3 OH OH H 3 C OH 3 69 70 71 72 H H3 C/ H 3 C H 3 C 73 74 75 76
H
3 0
H
3 0 77 78 79 80 -532- WO 99/54299 PCT/US99/07945
H
3 H3C H 3 C H 3 C 81 82 83 84 H3 3 3 H 3 C 85 86 87 88 H3 3 3 H 3 C 89 90 91 92 CH H 3 C
H
3 C CH 3
H
3 C CH 3
CH
3 CHa -.. CH 3
CH
3
CH
3
CH
3 93 94 95 96
CH
3 - H 3
CH
3 HCHC
CH
3
CH
3
CH
3
CH
3 97 98 99 100
CHH
3 C
H
3 C H 3 C 101 102 103 104
CH
3 105 106 107 108 -533- WO 99/54299 PCT/US99/07945
SOH
3 C HO , -CH 3 HO CH 3 HO,3 HO
OH
3
CH
3
OH
3 109 110 111 112
CH
3 HO -OH 3 HO OCH 3 HO HO
OH
3
OH
3
CH
3 113 114 115 116 OH CH H3C" H3C OH H3C OH OH HOH OH HO OH OH 117 118 119 120
SCH
3 OH H 3 C
H
3 C H 3 OH OOH HC OH OH OH OH 121 122 123 124 Table 4g CH 3 H3 C -OH3 H 3 c -OH 3 -H 3
-OH
3 OH OH OH OH 1 2 3 4 C H 3
CH
3
CH
3
CH
3
OH
3
H
3 OCk - -/H 3 0- ~J Q 9 OH OH OH OH 5 6 7 8 -534- WO 99/54299 PCT/US99/07945
H
3
-
C
H
3 3
CH
3 3 CH OH OH OH OH 9 10 11 12 CHI H3C H3 -A / [:A OH OH OH OH 13 14 15 16 - CH H 3 COA -CH 3
H
3 C -CH 3
-CH
3 -- H 3
OCH
3
OCH
3
OCH
3
OCH
3 17 18 19 20 S CH3 CH 3 CH3 CH3 CH3
H
3 CA --
H
3 0 -Ik-I
OH
3 k-i
OCH
3
OCH
3
OCH
3
OCH
3 21 22 23 24 S CH H 3 CJ CH3 OH 3 OH CH
OCH
3
OCH
3
OCH
3
OCH
3 25 26 27 28
CH
3 H3C --
H
3 ,A -=
OCH
3
OCH
3
OCH
3
OCH
3 29 30 31 32 H- CH 3 ^
H
3 CA-OH
H
3 0 -OH -OH -OH
OH
3
OH
3
OH
3
OH
3 33 34 35 36 H3O-OH
H
3 C -OH -OH -H 3 CH 3
OH
3
CH
3
OH
3 37 38 39 40 -535- WO 99/54299 PCT/US99/07945
H
3 C OH H 3 C0- -OH -OH U -O0H H3 C 3
CH
3
OH
3
OH
3
OH
3 41 42 43 44 H3 3 "--3
H
3c -OH
H
3 C OH -OH -OH 45 46 47 48
CH
3
H
3 -OCH3 H 3 C OCH 3 :- OCH 3
OCH
3
CH
3
OH
3
CH
3
OH
3 49 50 51 52
COH
3 C H H3 H3 3
CH
3
CH
3 H 3 53 54 55 56 ~CH
H
3 0 - H 3
H
3 C -OH 3 C 0cH 3
H
3
CH
3
CH
3
OH
3
CH
3 57 58 59 60
H
3 C-0CH 3
H
3
-H
3
-H
3 - H 3 61 62 63 64
CH
3 -H 3
OCH
3
OCH
3
-CH
3
-H
3 I OH \OH OH OH 65 66 67 68 -536- WO 99/54299 PCT/US99/07945
CH
3 HCH 3 CH
CH
3 CH 3 H OH OH OH 69 70 71 72
-CH
3
CH
3
CH
3 - CH 3 OH OH OH OH 73 74 75 76
CH
3 H O OH OH 77 78 79 80
CH
3 -OH 3
-CH
3
-CH
3
-H
3 0 OCH 3
OCH
3
\OCH
3
OCH
3 81 82 83 84
CH
3
CH
3
H
3 H 3
OCH
3
OCH
3
OCH
3
OCH
3 85 86 87 88
-CH
3
CH
3
OH
3 3CH 3 O CH 3 AOO CH 3 00O-H 3 00- H 3
OOH
3 89 90 91 92
CH
3
OCH
3
OCH
3
OCH
3
OCH
3 93 94 95 96
OH
3 -OH -OH OH -OH
OH
3 > OH 3 - OH 3
OH
3 97 98 99 100 -537- WO 99/54299 PCT/US99/07945 CH -OH -OH -OH H
CH
3
OH
3 CH 3 CH 3 101 102 103 104 CH -OH -OH -OH -OH
CH
3 CH 3 OH 3 CH 3 105 106 107 108
CH
3 OH -OH -OOH 109 110 111 112
CH
3
O
c
H
3
CH
3 1 CH 3
CH
3
H
3
OH
3
OH
3
OCH
3 113 114 115 116 CH H
CH
3
CH
3 3 CH 3 H 3
H
3
OH
3
CH
3
CH
3 117 118 119 120 CH Y -OOH 3 -O0H 3 -0 3 -00H 3
OH
3 OH 3 OH 3 OH 3 121 122 123 124
OH
3 Y ,-OCH 3 -O0H 3 30 -00H 3 125 126 127 128 -538- WO 99/54299 PCT/US99/07945 H3CH
H
3 O -kOH -OH I-OH -OH OH OH OH OH OHOH 129 130 131 132 H3H 3 3
-OCH
3 3 3 OH OH OH OH 133 134 135 136
H
3 C -OCH 3
-OCH
3
-HH
3 OCH OCH 3
OCH
3 OCH 137 138 139 140
CH
3
-I
H
3 C OH -OH -OH OH -OH OH OH -OH 141 142 143 144 %CH3
H
3 C -OCH 3 -,C, 3
OHC
3 3
OCH
3 -- OH oOH OH OH 145 146 147 148
OH
3
C
H
CH
3 CH"CH 3 C' -O 3H CH3 3 OCH 3 OCH 3
OH
3 OCH 3 149 150 151 152 -OH -O -O OH -OH OH OH 153 154 155 156 -539- WO 99/54299 PCT/US99/07945 SCH3 -OCH z -OCH 3 -O H 3 O H OH OH OH .OH 157 158 159 160
H
3 C -OCH3 H 3
H
3
-OCH
3
-OCH
3
CH
3
OCH
3
OCH
3
OCH
3
OCH
3 161 162 163 164 CH3
H
3 C OH H OH OH OH OH OH OH OH 165 166 167 168
CH
3 I
H
3 C -OCH 3
-OCH
3 -- OCH 3 OCH 3 OH OH OH OH 169 170 171 172 OCH
OCH
3 OCH 3 OCH 3 173 174 175 176
CH
3
H
3 C OH C OH OH OH 'OH -OH '-OH -OH '--OH 177 178 179 180
CH
3 H3C OCH 3 OCH 3 OCH3 H '-. OH '..-oH OOH 3
OOH
3 OH -OH '-OH -OH 181 182 183 184 -540- WO 99/54299 PCT/US99/07945 HC
OH
3
H
3 C OCH 3
OCH
3
OCH
3
OCH
3 • 3
-OCH
3 - OCH 3
-OCH
3
OC
3 185 186 187 188 OH OH OH OH OH '--OH -OH 'H-OH -OH 189 190 191 192
OCH
3
OCH
3
OCH
3 OCH OH -OH -OH '-OH 193 194 195 196
H
3 C OCH 3
OCH
3
OCH
3
OCH
3
H
3
-OCH
3 / "--OCH 3
-OCH
3
-OCH
3 CHz -OCH3 197 198 199 200 6$OH 6$OCH 3 OH OCH 3 201 202 203 204 OH OH OCH 3
OCH
3 205 206 207 208 OH OH OCH 3
OCH
3 209 210 211 212 -541- WO 99/54299 PCT/US99/07945 OH
OCH
3 O CH3OCHz 213 214 215 216 OCH3
CH
3 CH 3 CH 3 0- o0 (t :1 217 218 219 220 6 3CH 3
CCH
3
CH
3 221 222 223 224 The ability of the compounds of the invention to inhibit neuraminidase in vitro can be determined according to the method described below. Neuraminidase Inhibition Assay: Influenza virus AIN1/PRI8/34 was grown in the allantoic cavity of fertilized eggs and purified by sucrose density gradient centrifugation (Laver, W. G. (1969) in "Fundamental Techniques in Virology" (K. Habel and N. P. Salzman, eds.) pp. 92-86, Academic Press, New York). Influenza virus A/N2ITokyol3/67 was obtained from the tissue culture supernatents of virus grown on MDCK cells. Neuraminidase from B/Memphis/3/89 virus was prepared by digestion of the virus with TPCK-trypsin followed by centrifugation and then purification of the neuraminidase catalytic fragment using sucrose density gradient centrifugation and dialysis as described previously (Air, G. M., Laver, W. G., Luo, M., Stray, S. J., Legrone, G., and Webster, R. G. (1990) Viroloqy 177, 578-587). -542- WO 99/54299 PCT/US99/07945 The neuraminidase inhibition assays used the neuraminidase enzymatic activity associated with the A/N1/PR/8/34 or A/N2/Tokyo/3/67 whole virus, or the B/Memphis/3/89 catalytic head fragment. The whole virus or catalytic fragment was diluted appropriately with 20 mM N-ethylmorpholine, 10 mM. calcium choride, pH 7.5 buffer on the day of the experiment. Neuraminidase inhibition assays were conducted in 20 mM N-ethylmorpholine, 10 mM calcium choride, pH 7.5 buffer with 5% DMSO. Reaction mixtures included neuraminidase, inhibitor (test compound) and 20-30 ,M 4-methylumbelliferyl sialic acid substrate in a total volume of 200 pL and were contained in white 96-well U-shaped plates. Typically, five to eight concentrations of inhibitor were used for each Ki value measurement. The reactions were initiated by the addition of enzyme and allowed to proceed for 30-60 minutes at room temperature. The fluorescence for each well of the plate was measured once each minute during the reaction period by a Fluoroskan II plate reader (ICN Biomedical) equipped with excitation and emission filters of 355 +/- 35 nm and 460 +/- 25 nm, respectively. The plate reader was under the control of DeltaSoft II software (Biometallics) and a Macintosh computer. If the compound exhibited linear reaction velocities during the reaction period, then the reaction velocities for the dose-response study were fit to equation 1 using a nonlinear regression program (Kaleidagraph) to determine the overall Ki value (Segel, I. H. (1975) in Enzyme Kinetics, pp. 105-106, Wiley-lnterscience, New York). (1 - ViNo) = [1]/ {[I] + Ki(1 + [S]/Km)} eqn 1 In equation 1, Vi and Vo represent inhibited and uninhibited reaction velocities, respectively, and Km = 16 - 40,pM depending on the neuraminidase strain tested. For those compounds exhibiting slow-binding inhibition (Morrison, J. F. (1982) Trends Biochem. Sci. 7, 102-105), a second experiment was performed in a manner identical to the first except that neuraminidase and inhibitor were -543- WO 99/54299 PCT/US99/07945 preincubated in the absence of substrate for 2 hours at room temperature prior to initiating the reactions with substrate. Data analysis for the resulting linear velocities was conducted as described above. Equation 2 was used to measure Ki values in the sub-nanomolar range (Morrison, J. F. And Stone, S. R. (1985) Comments Mol. Cell Biophys. 2, 347 368). V = A{sqrt{(Ki' + It -Et)^2 + 4Ki'Et} - (Ki' + It - Et)] eqn. 2 In equation 2, V = velocity; A = akcat[S]/2(Km + [S]); a is a factor to convert fluorescence units to molar concentrations; Ki' = Ki(1 + [S]/Km); It = total inhibitor concentration and Et = total active concentration of neuraminidase. The compounds of the invention inhibit influenza A neuraminidase and influenza B neuraminidase with Ki values between about 0.1 nanomolar and about 500 micromolar. Preferred compounds of the invention invention inhibit influenza A neuraminidase and influenza B neuraminidase with Ki values between about 0.1 nanomolar and about 3.5 micromolar. The ability of the compounds of the invention to inhibit plaque formation in cell culture can be determined by the method described below. Cell Culture Plaque Formation Inhibition Assay Cell Cultures: MDCK cells obtained from the American Type Culture Collection were grown in Dulbecco's Modified Eagle Medium (DMEM) high glucose (GibcoBRL) supplemented with 10% fetal calf serum (JRH Biosciences), 40 mM HEPES buffer (GibcoBRL) and antibiotics (GibcoBRL). Cells were routinely cultured in flasks or roller bottles at 37 0 C and 5% CO 2 . At confluence cells were reduced to a density of 500,000 cells in a ml using trypsin/EDTA (GibcoBRL) treatment of the monolayer followed by cell centrifugation, resuspension, and -544- WO 99/54299 PCT/US99/07945 dilution into growth media. Cells were planted at a volume to surface area ratio of 1 ml over 1 cm 2 of growth surface. Plaque Assay Protocol: On MDCK cell confluent 6 well plates growth media was removed and the cells were overlaid with 1.5 ml of assay media (DMEM with 1% fetal calf serum, 40 mM HEPES buffer and antibiotics) containing pre-mixed virus (influenza A/Tokyo/3/67 [H2N2]) (40 -100 plaque forming units) and 2x concentration test compound. The plates were placed on a rocker and incubated for 2 hours at room temperature. During the virus adsorption period agar overlay media was prepared. In a microwave oven 2X agarose (final concentration of 0.6% agarose) in overlay media (DMEM with 40 mM HEPES buffer) was melted and then placed in a 48 0 C water bath for temperature equilibration. After the virus adsorption period was completed 1.5 ml agar over media was added and mixed with the 1.5 ml virus and test compound containing media per well. Cultures were incubated at 350C for the period required for plaque development, usually several days. Plaques were fixed with 3.7% formalin in PBS for 20 minutes followed by removal of the agar overlay and staining with 0.1% crystal violet in distilled water for 15 minutes. Plaques were counted and EC 50 concentration determined from multiple concentrations of the tested compound using regression analysis. Viral Stocks: Stocks were prepared in MDCK confluent roller bottles incubated at 370C in DMEM supplemented with 1% FCS, 40mM HEPES buffer, and antibiotics. Bottles were inoculated with a multiplicity of infection of approximately 0.1 plaque forming unit for each cell. Roller bottles were harvested after the cytopathic effect of the virus was observed to be complete. Stocks were prepared from the supernatant resulting from the low speed centrifugation of the media and cell lysate. Stocks were titered and stored at -800C. -545- WO 99/54299 PCT/US99/07945 Compounds of the invention provided plaque formation inhibition for influenza virus A/N2/Tokyo in MDCK cells with EC50 values between about 100 micromolar and about 1 nanomolar. Preferred compounds of the invention provided plaque formation inhibition for influenza virus A/N2/Tokyo in MDCK cells with EC50 values between about 1 micromolar and about 1 nanomolar. The compounds of the invention can be tested for in vivo antiviral activity using the method described below. In Vivo Antiviral Efficacy Method Female BALB/c mice were placed under anesthesia (sevoflurane) and inoculated intranasally (IN) with 0.1 ml of influenza A VR-95 (Puerto Rico PR8 34) at 10-2 (diluted from frozen stock). This viral concentration consistently produced disease in mice within 5 days of inoculation. Animals were treated 4h. pre-infection and 4h. post-infection, andperiodically thereafter, with one of the following therapies: no treatment; test compound (100, 25, 6.25, 1.39 mg/kg/day BID, PO); or vehicle (sterile water BID, PO). A group of ten animals (designated as control) was inoculated with 0.9% saline. Percent survival was determined. On day five, lungs were harvested, weighed and assigned scores of 0,1, 2, 3 or 4 based on percentage consolidation (0; 10-20; 25-50; 50-75; 75-100%, respectively). In addition, each lung pair was image analyzed to determine objective lung consolidation percentages. The compounds of the present invention can be used in the form of salts derived from inorganic or organic acids. These salts include but are not limited to the following: acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfonate (isethionate), lactate, maleate, methanesulfonate, nicotinate, 2 -546- WO 99/54299 PCT/US99/07945 naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3 phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, p toluenesulfonate and undecanoate. Also, basic nitrogen-containing groups can be quaternized with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chloride, bromides, and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl, and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides, and others. Water or oil-soluble or dispersible products are thereby obtained. Examples of acids which may be employed to form pharmaceutically acceptable acid addition salts include such inorganic acids as hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid and such organic acids as oxalic acid, maleic acid, succinic acid and citric acid. Other salts include salts with alkali metals or alkaline earth metals, such as sodium, potassium, lithium, calcium or magnesium or with ammonium or N(R**) 4 salts (where R** is loweralkyl). In addition, salts of the compounds of this invention with one of the naturally occurring amino acids are also contemplated. Preferred salts of the compounds of the invention include hydrochloride, methanesulfonate, sulfonate, phosphonate and isethionate. The compounds of the formula 1, II and III of this invention can have a substituent which is an acid group (for example, -CO 2 H, -SO 3 H, -SO 2 H, -PO 3
H
2 ,
-PO
2 H). Compounds of the formula 1, II and III of this invention having a substituent which is an ester of such an acidic group are also encompassed by this invention. Such esters may serve as prodrugs. The prodrugs of this invention are metabolized in vivo to provide the above-mentioned acidic substituent of the parental compound of formula 1, II or II. Prodrugs may also -547- WO 99/54299 PCT/US99/07945 serve to increase the solubility of these substances and/or absorption from the gastrointestinal tract. These prodrugs may also serve to increase solubility for intravenous administration of the compounds. Prodrugs may also serve to increase the hydrophobicity of the compounds. Prodrugs may also serve to increase the oral bioavailability of the compounds by increasing absorption and/or decreasing first-pass metabolism. Prodrugs may also serve to increase tissue penetration of the compounds, thereby leading to increased activity in infected tissues and/or reduced rate of clearance. Such esters contemplated by this invention include: alkyl esters, especially loweralkyl esters, including, but not limited to, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, t-butyl, n-pentyl esters and the like; alkoxyalkyl esters, especially, loweralkoxyloweralkyl esters, including, but not limited to, methoxymethyl, 1-ethoxyethyl, 2-methoxyethyl, isopropoxymethyl, t-butoxymethyl esters and the like; alkoxyalkoxyalkyl esters, especially, alkoxyalkoxy-substituted loweralkyl esters, including, but not limited to, 2-methoxyethoxymethyl esters and the like; aryloxyalkyl esters, especially, aryloxy-substituted loweralkyl esters, including, but not limited to, phenoxymethyl esters and the like, wherein the aryl group is unsubstituted or substituted as previously defined herein; haloalkoxyalkyl esters, especially, haloalkoxy-substituted loweralkyl esters, including, but not limited to, 2,2,2-trichloroethoxymethyl esters and the like; alkoxycarbonylalkyl esters, especially, loweralkoxycarbonyl-substituted loweralkyl esters, including, but not limited to, methoxycarbonylmethyl esters and the like; -548- WO 99/54299 PCT/US99/07945 cyanoalkyl esters, especially, cyano-substituted loweralkyl esters, including, but not limited to, cyanomethyl, 2-cyanoethyl esters and the like; thioalkoxymethyl esters, especially, lowerthioalkoxy-substituted methyl esters, including, but not limited to, methylthiomethyl, ethylthiomethyl esters and the like; alkylsulfonylalkyl esters, especially, loweralkylsulfonyl-substituted loweralkyl esters, including, but not limited to, 2-methanesulfonylethyl esters and the like; arylsulfonylalkyl esters, especially, arylsulfonyl-substituted loweralkyl esters, including, but not limited to, 2-benzenesulfonylethyl and 2 toluenesulfonylethyl esters and the like; acyloxyalkyl esters, especially, loweralkylacyloxy-substituted loweralkyl esters, including, but not limited to, formyloxymethyl, acetoxymethyl, pivaloyloxymethyl, acetoxyethyl, pivaloyloxyethyl esters and the like; cycloalkylcarbonyloxyalkyl esters including, but not limited to, cyclopentanecarbonyloxymethyl, cyclohexanecarbonyloxymethyl, cyclopentanecarbonyloxyethyl, cyclohexanecarbonyloxyethyl esters and the like; arylcarbonyloxyalkyl esters including, but not limited to, benzoyloxymethyl esters and the like; (alkoxycarbonyloxy)alkyl esters, especially, (loweralkoxycarbonyloxy) substituted loweralkyl esters, including, but not limited to, methoxycarbonyloxymethyl, ethoxycarbonyloxymethyl, 1 (methoxycarbonyloxy)ethyl, 2-(ethoxycarbonyloxy)ethyl esters and the like; (cycloalkyloxycarbonyloxy)alkyl esters, especially, (cycloalkyloxycarbonyloxy)-substituted loweralkyl esters, including, but not limited -549- WO 99/54299 PCT/US99/07945 to, cyclohexyloxycarbonyloxymethyl, cyclopentyloxycarbonyloxyethyl, cyclohexyloxycarbonyloxypropyl esters and the like; oxodioxolenylmethyl esters including, but not limited to, (5-phenyl-2-oxo 1,3-dioxolen-4-yl)methyl, [5-(4-methylphenyl)-2-oxo-1,3-dioxolen-4-yl]methyl, [5 (4-methoxyphenyl)-2-oxo-1,3-dioxolen-4-yl]methyl, [5-(4-fluorophenyl)-2-oxo-1,3 dioxolen-4-yl]methyl, [5-(4-chlorophenyl)-2-oxo-1,3-dioxolen-4-yl]methyl, (2-oxo 1,3-dioxolen-4-yl)methyl, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl, (5-ethyl-2 oxo-1,3-dioxolen-4-yl)methyl, (5-propyl-2-oxo-1,3-dioxolen-4-yl)methyl, (5 isopropyl-2-oxo-1,3-dioxolen-4-yl)methyl, (5-butyl-2-oxo-1,3-dioxolen-4-yl)methyl esters and the like; phthalidyl esters wherein the phenyl ring of the phthalidyl group is unsubstituted or substituted as defined previously herein, including, but not limited to, phthalidyl, dimethylphthalidyl, dimethoxyphthalidyl esters and the like; aryl esters including, but not limited to, phenyl, naphthyl, indanyl esters and the like; arylalkyl esters, especially, aryl-substitued loweralkyl esters, including, but not limited to, benzyl, phenethyl, 3-phenylpropyl, naphthylmethyl esters and the like, wherein the aryl part of the arylalkyl group is unsubstituted or substituted as previously defined herein; dialkylaminoalkyl esters, especially dialkylamino-substituted loweralkyl esters, including, but not limited to, 2-(N,N-dimethylamino)ethyl, 2-(N,N diethylamino)ethyl ester and the like (heterocyclic)alkyl esters, especially, heterocyclic-substituted loweralkyl esters wherein the heterocycle is a nitrogen-containing heterocycle, including, but not limited to, (heterocyclic)methyl esters and the like, wherein the heterocyclic part of the (heterocyclic)alkyl group is unsubstituted or substituted as previously defined herein; and -550- WO 99/54299 PCT/US99/07945 carboxyalkyl esters, especially, carboxy-substituted loweralkyl esters, including, but not limited to carboxymethyl esters and the like; and the like. Preferred prodrug esters of acid-containing compounds of the Formula 1, II or III are loweralkyl esters, including, but not limited to, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, t-butyl, n-pentyl esters and benzyl esters wherein the phenyl ring is unsubstituted or substituted as previously defined herein. Methods for the preparation of prodrug esters of compounds of the Formula 1, II or III are well-known in the art and include: reacting the acid with the corresponding halide (for example, chloride or acyl chloride) and a base (for example, triethylamine, DBU, N,N dimethylaminopyridine and the like) in an inert solvent (for example, DMF, acetonitrile, N-methylpyrrolidone and the like); reacting an activated derivative of the acid (for example, an acid chloride, sulfonyl chloride, monochlorophosphonate and the like) with the corresponding alcohol or alkoxide salt; and the like. Other examples of prodrugs of the present invention include esters of hydroxyl-substituted compounds of formula I, II or III which have been acylated with a blocked or unblocked amino acid residue, a phosphate function, a hemisuccinate residue, an acyl residue of the formula RloC(O)- or RIoC(S) wherein R 1 00 is hydrogen, lower alkyl, haloalkyl, alkoxy, thioalkoxy, alkoxyalkyl, thioalkoxyalkyl or haloalkoxy, or an acyl residue of the formula Ra-C(Rb)(Rd)-c(O)- or Ra-C(Rb)(Rd)-C(S) - wherein Rb and Rd are independently selected from hydrogen or lower alkyl and R a is -N(Re)(Rf), -ORe or -SR e wherein Re and R f are independently selected from hydrogen, lower alkyl and haloalkyl, or an amino-acyl residue having the formula Rio'NH(CH 2
)
2
NHCH
2 C(O)- or -551- WO 99/54299 PCT/US99/07945 Ri'NH(CH 2
)
2
OCH
2 C(O)- wherein R 1
'
01 is hydrogen, lower alkyl, (aryl)alkyl, (cycloalkyl)alkyl, acyl, benzoyl or an a-amino acyl group. The amino acid esters of particular interest are of glycine and lysine; however, other amino acid residues can also be used, including any of the naturally occuring amino acids and also including those wherein the amino acyl group is -C(O)CH 2
NR
02
R
10 3 wherein R 1 0 2 and R 1 0 3 are independently selected from hydrogen and lower alkyl, or the group
-NR
1 02
R
103 , where R 1 02 and R 1 03 , taken together, forms a nitrogen containing heterocyclic ring. Other prodrugs include a hydroxyl-substituted compound of formula I, II or Ill wherein the hydroxyl group is functionalized with a substituent of the formula -CH(R'4)OC(O)Ri' 0 5 or -CH(R 104
)OC(S)R
10 5 wherein R 105 is lower alkyl, haloalkyl, alkoxy, thioalkoxy or haloalkoxy and R 1 04 is hydrogen, lower alkyl, haloalkyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl or dialkylaminocarbonyl. Such prodrugs can be prepared according to the procedure of Schreiber (Tetrahedron Lett. 1983, 24, 2363) by ozonolysis of the corresponding methallyl ether in methanol followed by treatment with acetic anhydride. The preparation of esters of hydroxyl-substituted compounds of formula formula I, II or III is carried out by reacting a hydroxyl-substituted compound of formula formula I, II or III, with an activated amino acyl, phosphoryl, hemisuccinyl or acyl derivative. Prodrugs of hydroxyl-substituted-compounds of the invention can also be prepared by alkylation of the hydroxyl substituted compound of formula formula I, II or III, with (halo)alkyl esters, transacetalization with bis-(alkanoyl)acetals or condensation of the hydroxyl group with an activated aldehyde followed by acylation of the intermediate hemiacetal. In preparing prodrugs it often is necessary to protect other reactive functional groups, in order to prevent unwanted side reactions. After protection of -552- WO 99/54299 PCT/US99/07945 the reactive groups the desired group can be functionalized. The resulting functionalized product is then deprotected, to remove the protecting groups that were added to prevent unwanted side reactions. This will provide the desired prodrug. Suitable reaction conditions for preparing protecting groups are well known in the art. One source for reaction conditions is found in T.H. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 2nd edition, John Wiley & Sons, New York (1991). This invention also encompasses compounds of the Formula 1, II or III which are esters or prodrugs and which are also salts. For example, a compound of the invention can be an ester of a carboxylic acid and also an acid addition salt of an amine or nitrogen-containing substituent in the same compound. The compounds of the present invention are useful for inhibiting neuraminidase from disease-causing microorganisms which comprise a neuraminidase. The compounds of the invention are useful (in humans, other mammals and fowl) for treating or preventing diseases caused by microorganisms which comprise a neuraminidase The compounds of the present invention are useful for inhibiting influenza A virus neuraminidase and influenza B virus neuraminidase, in vitro or in vivo (especially in mammals and, in particular, in humans). The compounds of the present invention are also useful for the inhibition of influenza viruses, orthomyxoviruses, and paramyxoviruses in vivo, especially the inhibition of influenza A viruses and influenza B viruses in humans and other mammals. The compounds of the present invention are also useful for the treatment of infections caused by influenza viruses, orthomyxoviruses, and paramyxoviruses in vivo, especially the human diseases caused by influenza A and influenza B viruses. The compounds of the present invention are also useful for the prophylaxis of infections caused by influenza viruses, orthomyxoviruses, and paramyxoviruses in vivo in humans and other mammals, especially the prophylaxis of influenza A -553- WO 99/54299 PCT/US99/07945 and influenza B viral infections; and, in particular, the prophylaxis of influenza A and influenza B viral infections in human subjects who are at high risk of developing other respiratory diseases concurrent with or as a consequence of influenza virus infections, or who suffer from chronic respiratory illness, such as asthma, emphysema, or cystic fibrosis. Total daily dose administered to a human or other mammal host in single or divided doses may be in amounts, for example, from 0.001 to 300 mg/kg body weight daily and more usually 0.1 to 10 mg/kg body weight daily. Dosage unit compositions may contain such amounts of submultiples thereof to make up the daily dose. 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. It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination, and the severity of the particular disease undergoing therapy. Administration of a compound of this invention will begin before or at the time of infection or after the appearance of established symptoms and/or the confirmation of infection. The compounds of the present invention may be administered orally, parenterally, sublingually, intranasally, by intrapulmonary administration, by inhalation or insufflation as a solution, suspension or dry powder (for example, in a spray), or rectally, in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired. The -554- WO 99/54299 PCT/US99/07945 term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection, or infusion techniques. Injectable preparations, for example, sterile injectable aqueous or oleagenous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-propanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. Suppositories for rectal administration of the drug can be prepared by mixing the drug with a suitable nonirritating excipient such as cocoa butter and polyethylene glycols which are solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum and release the drug. Solid dosage forms for oral administration may include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound may be admixed with at least one inert diluent such as sucrose lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., lubricating agents such as magnesium stearate. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents. Tablets and pills can additionally be prepared with enteric coatings. Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert -555- WO 99/54299 PCT/US99/07945 diluents commonly used in the art, such as water. Such compositions may also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents. The compounds of the present invention can also be administered in the form of liposomes. As is known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by mono- or multi-lamellar hydrated liquid crystals that are dispersed in an aqueous medium. Any non-toxic, physiologically acceptable and metabolizable lipid capable of forming liposomes can be used. The present compositions in liposome form can contain, in addition to a compound of the present invention, stabilizers, preservatives, excipients, and the like. The preferred lipids are the phospholipids and phosphatidyl cholines (lecithins), both natural and synthetic. Methods to form liposomes are known in the art. See, for example, Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, N.Y. (1976), p. 33 et seq. While the compounds of the invention can be administered as the sole active pharmaceutical agent, they can also be used in combination with one or more anti-infective agents and/or other agents used to treat other acute or chronic respiratory ailments. Other agents to be administered in combination with a compound of the present invention include: an influenza vaccine; other influenza inhibitors such as, for example, amantadine, rimantadine, ribavirin, and the like; another influenza neuraminidase inhibitor, such as, for example, zanamivir or GS 4104 and the like; agents used to treat respiratory bacterial infections and bronchitis, such as, for example, erythromycin, clarithromycin, azithromycin and the like; and agents used to treat asthma, such as, for example, zileuton, albuterol (salbutamol), salmeterol, formoterol, ipratropium bromide, inhaled steroids and the like, or anti-inflammatory agents for treating asthma such as, for example, beclomethasone dipropionate, fluticasone propionate, -556- WO 99/54299 PCT/US99/07945 budesonide, triamcinolone acetonide, flunisolide, cromolyn, zafirlukast, montelukast used in combination with a compound of the present invention. When administered as a combination, the therapeutic agents can be formulated as separate compositions which are given at the same time or different times, or the therapeutic agents can be given as a single composition. The foregoing is merely illustrative of the invention and is not intended to limit the invention to the disclosed compounds. Variations and changes which are obvious to one skilled in the art are intended to be within the scope and nature of the invention which are defined in the appended claims. -557-
Claims (56)
1. A compound of the formula: Y R9 R8 Rio R 6 R 2 -X N1 NN R ' R R Rs R 3 or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein R 1 is selected from the group consisting of (b) -CO 2 H, (b) -CH 2 CO 2 H, (c) -SO 3 H, (d) -CH 2 SO 3 H, (e) -SO 2 H, (g) -CH 2 SO 2 H, (g) -P0 3 H 2 , (h) -CH 2 PO 3 H 2 , (i) -PO 2 H, (j) -CH 2 PO 2 H, (I) tetrazolyl, (I) -CH 2 -tetrazolyl, (m) -C(=0)-NH-S(O) 2 -R 11 , (0) -CH 2 C(=O)-NH-S(O) 2 -R 11 , (0) -SO 2 N(T-R 11 )R 12 and (p) -CH 2 SO 2 N(T-R 1 )R 12 wherein T is selected from the group consisting of (i) a bond, (ii) -C(=O)-, (iii) -C(=O)O-, (iv) -C(=O)S-, (v) -C(=O)NR 36 -, (vi) -C(=S)O-, (vii) -C(=S)S-, and (viii) -C(=S)NR 36 -, R 11 is selected from the group consisting of (i) C1-C 1 2 alkyl, (ii) C 2 -C 12 alkenyl, (iii) cycloalkyl, (iv) (cyclo alkyl)alkyl, (w) (cycloalkyl)alkenyl, (vi) cycloalkenyl, (vii) (cycloalkenyl)alkyl, (ix) (cycloalkenyl)alkenyl, (ix) aryl, (x) (aryl)alkyl, (xi) (aryl)alkenyl, -558- WO 99/54299 PCT/US99/07945 (xvii) heterocyclic, (xiii) (heterocyclic)alkyl and (xviii) (xiv) (heterocyclic)alkenyl; and R 12 and R 36 are independently selected from the group consisting of (i) hydrogen, (ii) Cl-C12 alkyl, (iii) C2-C12 alkenyl, (iv) cycloalkyl, (v) (cycloalkyl)alkyl, (vi) (cycloalkyl)alkenyl, (vii) cycloalkenyl, (viii) (cycloalkenyl)alkyl, (ix) (cycloalkenyl)alkenyl, (x) aryl, (xi) (aryl)alkyl, (xii) (aryl)alkenyl, (xiii) heterocyclic, (xiv) (heterocyclic)alkyl and (xv) (heterocyclic)alkenyl; X is selected from the group consisting of (a) -C(=O)-N(R*)-, (b) -N(R*)-C(=O)-, (c) -C(=S)-N(R*)-, (d) -N(R*)-C(=S)-, (e) -N(R*)-SO 2 -, and (f) -SO 2 -N(R*)- wherein R* is hydrogen, C 1 -C 3 loweralkyl or cyclopropyl; R 2 is selected from the group consisting of (a) hydrogen, (b) C 1 -C 6 alkyl, (c) C2-C6 alkenyl, (d) C3-C6 cycloalkyl, (e) Cs-C6 cycloalkenyl, (f) halo C1-C6 alkyl and (g) halo C2-C6 alkenyl; or R 2 -X- is Y 1 2 ( /,y/N- Y wherein Y 1 is -CH 2 -, -0-, -S- or -NH- and Y 2 is -C(=O)- or -C(Raa)(Rbb) - wherein Raa and Rbb are indepedently selected from the group consisting of hydrogen, C1-C3 loweralkyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, aminomethyl, 1-aminoethyl, 2-aminoethyl, thiolmethyl, 1-thiolethyl, 2-thiolethyl, methoxymethyl, N-methylaminomethyl and methylthiomethyl; R 3 and R 4 are independently selected from the group consisting of (a) hydrogen, (b) cycloalkyl, (c) cycloalkenyl, (d) heterocyclic, (e) aryl and -559- WO 99/54299 PCTIUS99/07945 ()-Z-R 14 wherein Z is (v) -C(=S)-, (vi) -C(=NR 1 5 )-, (vii) -C(R 37 a) (OR 7 c)_, (Viii) C(R 37 a) (SR 37 c)_, (ix) -C(R 37a) (N(R 37 b) (R 37 c))-, (x -C(R 37 a)( R 37 b)-o-, (xi) -C(R 37a)(R 37 b )-N(R 37 c)_, (xii) -C(R 37a)(R 37 b )-N(O)(R 37 c)_, (xiii) -C(R 37 a)(R 37 b )-N(OH)-, (xiv) -O(R 37 a)( R 37 b)_S_, (xv) -C(R 37 a)(R 37 b)-S(O)-, (xvi) -C(R 37 a)(R 37 b)-S(O) 2 -, (xviii) -C(R 3 7 a)( R 37 b)_CQ=O)_, (xviii) -C(R 37 1)(R 37 b)C(=S)-, (xxi) -C(R 37 a)(R 37 b)_C(=NR R 1 5)-, (xx) -C(R 37 a) (OR 37 C)-C(=O)-, (xxi) -C(R 37 a) (SR 37 c)_C(=O)_, (xxii) -C(R 37 a)(O R 37 c)c (=S)-, (xxiii) -C(R 37 a)(SR 37 c)_C(=S)_, (xxiv) -C(=O)-C(R 37 a)(OR 37 c)-, (xxv) -C(=O)-C(R 37 a)(S R 37 c)_, (xxvi) -O(=S)-C(R 37 a)(O R 37 c)-, (xxvii) -C(=S)-C(R 37 a)(SR 37 c)_, (xxviii) -C (R 37 a)(O R3 7 c)_C(R 37 a)(OR 37 c)-, (xxix) -C(R 37a)(SR 37 c)_C (R 37 a) (OR R 3 7C)_, (xxx) -C(R 37a) (OR 37c)_C (R 37 a) (SR 3 C, (xxxi) -C(R 37a)(SR 37 rc)_C(R 37 a)(SR z) (xxxii) -C(=O)-C(0O)-, (xxxiii) -C(=S)-C(=S)-, (xxxiv) -C(0)-O-. (xxxv) -C(0)-S-, (xxxvi) -C(=S)-O-, (xxxvii) -C(=S)-S-, (xxxviii) -C(=O)-N(R 37 a)_, (xxxix) -C(=S)-N(R 37 a)_, (Xl) -C(R 3a)(R 37b,)_C(=O)-N(R 37 a)_, -560- WO 99/54299 PCT/US99/07945 (xli) -C(R 37 a) (R 37 b)_C (=S)-N ( R 37 a)_, (xiii) -C(R 37 a) (R 37 b)_C(=O0)-O-, (xlv) -C(R 37 a) (R 37 b)_C(=S)_S_, (xlvi) -C(R 37 a) (R 37 b )-N(R 37 b)-c(=o)-, (x lix ) C (R 3 7 a )(R 3 7 b ) S C (= O ) , (I) C (R 3 7 a )(R 3 7 b ) O C (= S ) , R 3 a (liii) -CR3a( 7)NR37)C=)NR3a_ (liv) -CR3a)R3bN( 7) (O- (lv) -CR3a R3b-( 7)c(O- (lvi) -C(R 37 a) (R 37 b)-N (R 37 b)_C(=S)-O-, (lvii) -CR3a R3b)NR3b_(S__ (lviii) -CR3a( 7)oC=)NR3a_ (lix) -CR31( 7)SC=)NR3a_ (lxv) -C( (l)( 7b_-(O)S xvi) -CR3a R1b__(S__ (lxix) -CR3a R37)SC=)S or (lxx) -C(R 37 a) (R 37 b)_ C(R 37 a) (OR 37 c)-; R 1 4 ijS (i) hydrogen, (ii) 0 1 -C 12 alkyl, (iii) haloalkyl, (iv) hydroxyalkyl, -561- WO 99/54299 PCT/US99/07945 (v) thiol-substituted alkyl, (vi) R 37 cO-substituted alkyl, (vii) R 3 7 cS-substituted alkyl, (viii) aminoalkyl, (ix) (R 37 c)NH-substituted alkyl, (x) (R 37 a)(R 37 c)N-susbstituted alkyl, (xi) R 37 aO-(O=)C-substituted alkyl, (xii) R 37 aS-(O=)C-substituted alkyl, (xiii) R3 7 aO-(S=)C-substituted alkyl, (xix) R 37 aS-(S=)C-substituted alkyl, (xx) (R 37 aO) 2 -P(=O)-substituted alkyl, (xvi) cyanoalkyl, (xxi) C2-C12 alkenyl, (xviii) haloalkenyl, (xix) C2-C12 alkynyl, (xxii) cycloalkyl, (xxi) (cycloalkyl)alkyl, (xxii) (cycloalkyl)alkenyl, (xxiv) (cycloalkyl)alkynyl, (xxiv) cycloalkenyl, (xxv) (cycloalkenyl)alkyl, (xxvii) (cycloalkenyl)alkenyl, (xxvii) (cycloalkenyl)alkynyl, (xxviii) aryl, (xxx) (aryl)alkyl, (xxx) (aryl)alkenyl, (xxxi) (aryl)alkynyl, (xxxii) heterocyclic, (xxxiii) (heterocyclic)alkyl, (xxxiv) (heterocyclic)alkenyl or (xxxv) (heterocyclic)alkynyl, with the proviso that R 1 4 is other than hydrogen when Z is -C(R 37 a)(R 3 7 b) - N (R 3 7 b)-C(=O)-O-, -C(R 3 7 a)(R 37 b)-N ( R 37 b)-C(=S)-O-, -C(R37a)(R37b)-N(R37b)-C(=O)-S -, -C(R37a)(R37b)-N(R37b)-C(=S)-S - , -C(R37a)(R37b)-o-C(=O)-o - , -C(R37a)(R37b)-O-C(=S)-O - , -C(R37a)(R37b)_-SC(=O)-o - , -C(R37a)(R37b)-S-C(=S)-O - , -562- WO 99/54299 PCT/US99/07945 -C(R37a)(R37b)_oC(=O)-S - , -C(R37a)(R37b)-O-C(=S)-S - , -C(R37a)(R37b)-S-C(=O)-S - or -C(R 37 a)(R 37 b)-S-C(=S)-S-; R 37 a, R 37 b, R 4 7 , and R 48 at each occurrence are independently selected from the group consisting of (i) hydrogen, (ii) C1-C12 alkyl, (iii) haloalkyl, (iv) hydroxyalkyl, (v) alkoxyalkyl, (vi) C2-C12 alkenyl, (vii) haloalkenyl, (viii) C2-012 alkynyl, (ix) cycloalkyl, (x) (cycloalkyl)alkyl, (xi) (cycloalkyl)alkenyl, (xii) (cycloalkyl)alkynyl, (xiii) cycloalkenyl, (xiv) (cycloalkenyl)alkyl, (xv) (cycloalkenyl) alkenyl, (xvi) (cycloalkenyl)alkynyl, (xvii) aryl, (xviii) (aryl)alkyl, (xix) (aryl)alkenyl, (xx) (aryl)alkynyl, (xxi) heterocyclic, (xxii) (heterocyclic)alkyl, (xxiii) (heterocyclic)alkenyl and (xxiv) (heterocyclic)alkynyl; R 37 c at each occurrence is independently selected from the group consisting of (i) hydrogen, (ii) C1-C12 alkyl, (iii) haloalkyl, (iv) C2-C12 alkenyl, (v) haloalkenyl, (vi) C2-C12 alkynyl, (vii) cycloalkyl, (viii) (cycloalkyl)alkyl, (ix) (cycloalkyl)alkenyl, (x) (cycloalkyl)alkynyl, (xi) cycloalkenyl, (xii) (cycloalkenyl)alkyl, (xiii) (cycloalkenyl)alkenyl, -563- WO 99/54299 PCT/US99/07945 (xiv) (cycloalkenyl)alkynyl, (xv) aryl, (xvi) (aryl)alkyl, (xvii) (aryl)alkenyl, (xviii) (aryl)alkynyl, (xix) heterocyclic, (xx) (heterocyclic)alkyl, (xxi) (heterocyclic)alkenyl, (xxiv) (heterocyclic)alkynyl, (xxiii) -C(=O)-R 14 , (xxiv) -C(=S)-R 4 , (xxv) -S(O) 2 -R 1 4 and (xxvi) hydroxyalkyl; or when Z is -C(R 37 )(R 37 b)-N(R37c)-, then N(R 37 c) and R 1 4 when taken together are an azido group; or when Z is -C(R 3 7 a)(R 37 b)-N(O)(R 37 c) - , then N(O)(R 37 c) and R 1 4 when taken together are an N-oxidized 3-7 membered heterocyclic ring having at least one N-oxidized ring nitrogen atom; or when Z is -C(R 3 7 a)(OR 3 7 c) - , -C(R 37 a)(SR 37 c) - or -C(R 37 a)(N(R 37 b)(R 37 c)) - , then R 37a , R 1 4 and the carbon atom to which they are bonded when taken together form a cyclopentyl, cyclopentenyl, cyclohexyl or cyclohexenyl ring; R i s 5 is selected from the group consisting of (i) hydrogen, (ii) hydroxy, (iii) amino, (iv) C1-C12 alkyl, (v) haloalkyl, (vi) C2-C12 aikenyl, (vii) haloalkenyl, (viii) cycloalkyl, (ix) (cycloalkyl)alkyl, (x) (cycloalkyl)alkenyl, (xi) cycloalkenyl, (xii) (cycloalkenyl)alkyl, (xiii) (cycloalkenyl)alkenyl, (xiv) aryl, (xv) (aryl)alkyl, (xvi) (aryl)alkenyl, (xvii) heterocyclic, (xviii) (heterocyclic)alkyl and (xix) (heterocyclic)alkenyl; -564- WO 99/54299 PCT/US99/07945 or R 3 and R 4 taken together, with the atom to which they are attached, form a carbocyclic or heterocyclic ring having from 3 to 8 ring atoms; R 5 is selected from the group consisting of (a) hydrogen, (b) -CH(R 38 ) 2 , (c) -O-R 40 , (d) C2-C4 alkynyl, (e) cyclopropyl, (f) cyclobutyl, (g) -C(=Q 1 )-R 17 , and (h) -N(R 19 ) 2 wherein Q 1 is O, S, or N(R18); R 17 and R 1 8 are independently selected, at each occurrence, from the group consisting of hydrogen, methyl, and ethyl; R' 9 , R 3 8 , and R 4 0 are independently selected, at each occurrence, from the group consisting of (i) hydrogen, (ii) C01-C12 alkyl, (iii) haloalkyl, (iv) C2-C12 alkenyl, (v) haloalkenyl, (vi) cycloalkyl, (vii) (cycloalkyl)alkyl, (viii) (cycloalkyl)alkenyl, (ix) cycloalkenyl, (x) (cycloalkenyl)alkyl, (xi) (cycloalkenyl)alkenyl, (xii) aryl, (xiii) (aryl)alkyl, (xiv) (aryl)alkenyl, (xv) heterocyclic, (xvi) (heterocyclic)alkyl and (xvii) (heterocyclic)alkenyl; Y is selected from the group consisting of (a) hydrogen, (b) C1-C5 alkyl, (c) Cj-C5 haloalkyl, (d) C2-C5 alkenyl, (e) C2-05 haloalkenyl, (f) C2-05 alkynyl, (g) C3-05 cycloalkyl, (h) C3-C5 cycloalkyl-Cl-to-C 3 -alkyl, (i) C5 cycloalkenyl, (j) C5 cycloalkenyl-Cl-to C 3 -alkyl, (k) C5 cycloalkenyl-C 2 -to-0 3 -alkenyl, (I) -(CHR 39 )nOR 20 , (m) -CH(OR 20 ) -565- WO 99/54299 PCT/US99/07945 CH 2 (OR 20 ), (n) -(CHR 39 )nSR 21 , (0) -(CHR 3 9 )nCN, (p) -(CHR3 9 )nN 3 , (q) phenyl, (r) halo-substituted phenyl, (s) -(CHR 39 )nC(=Q 2 )R 22 , (t) -(CHR 39 )nN(=Q 3 ), (u) -N(O)=CHCH 3 , (v) -(CHR 39 )nNR 23 R 24 , (w) halo and (x) a heterocyclic ring having from 3 to 6 ring atoms; wherein n is 0, 1, or 2; Q 2 is O, S, NR 25 , or CHR 26 ; and Q 3 is NR 41 , or CHR 4 2 ; R 20 at each occurrence is independently (i) hydrogen, (ii) methyl, (iii) ethyl, (iv) n-propyl, (v) isopropyl, (vi) C1-C3 haloalkyl, (vii) vinyl, (viii) propenyl, (ix) isopropenyl, (x) allyl, (xi) C2-C3 haloalkenyl, (xii) amino, (xiii) -NHCH 3 , (xiv) -N(CH 3 ) 2 , (xv) -NHCH 2 CH 3 , (xvi) -N(CH 3 )(CH 2 CH 3 ), (xvii) -N(CH 2 CH 3 ) 2 or (xviii) -N(=CH 2 ); R 2 1 is hydrogen, (ii) methyl, (iii) ethyl, (iv) n-propyl, (v) isopropyl, (vi) C1-C3 haloalkyl, (vii) vinyl, (viii) propenyl, (ix) isopropenyl, (x) allyl or (xi) C2-C3 haloalkenyl; R 22 is (i) hydrogen, (ii) methyl, (iii) ethyl, (iv) n-propyl, (v) isopropyl, (vi) hydroxy, (vii) thiol, (viii) methoxy, (ix) ethoxy, (x) n-propoxy, (xi) isopropoxy, (xii) cyclopropyloxy, (xiii) methylthio, (xiv) ethylthio, (xv) n-propylthio, (xvi) isopropylthio, (xvii) cyclopropylthio, (xviii) vinyl, (xix) propenyl, (xx) isopropenyl, (xxi) allyl, (xxii) -N(R28a)(R 28 b), (xxv) -CH 2 R 29 , (xxiv) aminomethyl, (xxv) hydroxymethyl, -566- WO 99/54299 PCT/US99/07945 (xxvi) thiolmethyl, (xxvii) -NHNH 2 , (xxviii) -N(CH 3 )NH 2 or (xxix) -NHNH(CH 3 ); R 2 3 and R 3 9 are independently hydrogen or methyl; R 41 and R 4 2 are independently hydrogen, methyl, or ethyl; R 24 is selected from the group consisting of (i) hydrogen, (ii) C1-C4 alkyl, (iii) C2-C4 alkenyl, (iv) C2-C4 alkynyl, (v) cyclopropyl, (vi) -C(=Q 4 )-R 30 , (v) -OR 31 , and (vi) -N(R 32 ) 2 , wherein Q 4 is O, S, or N(R 33 ); R 25 is hydrogen, hydroxy, methyl, ethyl, amino, -CN, or -NO 2 ; R 2 6 group is hydrogen, methyl or ethyl; R 28 a hydrogen, hydroxy, methyl, ethyl, amino, -NHCH 3 , -N(CH 3 ) 2 , methoxy, ethoxy, or -CN; R 28 b is hydrogen, methyl or ethyl; or R 28 a , R 2 8 b and the nitrogen to which they are bonded taken together represent azetidinyl; R 29 group is hydrogen, hydroxy, thiol, methyl, ethyl, amino, methoxy, ethoxy, methylthio, ethylthio, methylamino or ethylamino; R 30 group is hydrogen, methyl, ethyl, -OR 34 , -SR 34 , -N(R 35 ) 2 , -NHOH, -NHNH 2 , -N(CH 3 )NH 2 , or -N(CH 2 CH 3 )NH2; R 31 and R 3 2 substituents, at each occurrence, are independently hydrogen, methyl or ethyl; R 3 3 group is hydrogen, hydroxy, methyl, ethyl, amino, -CN, or -NO 2 ; -567- WO 99/54299 PCT/US99/07945 R34 group is methyl or ethyl; R 35 group is independently hydrogen, methyl or ethyl; with the proviso that when Q 2 is CHR 2 6 then R 22 is selected from the group consisting of hydrogen, -CH 3 , -C 2 H 5 , -C 3 H 7 , -OCH 3 , -SCH 3 , -O-0 2 H 5 , and -S-C 2 H 5 , and with the proviso that when R 3 and R 4 are each hydrogen, then Y is other than hydrogen; R 6 and R 7 are independently selected from the group consisting of (c) hydrogen, (b) C1-C12 alkyl, (c) C2-012 alkenyl, (d) cycloalkyl, (e) (cycloalkyl)alkyl, (f) (cycloalkyl)alkenyl, (g) cycloalkenyl, (h) (cyclo alkenyl)alkyl, (j) (cycloalkenyl)alkenyl, (j) aryl, (k) (aryl)alkyl, (I) (aryl)alkenyl, (m) heterocyclic, (o) (heterocyclic)alkyl and (0) (heterocyclic)alkenyl; and R 8 , R 9 , and R 1 0 are independently selected from the group consisting of (a) hydrogen, (b) C1-C6 alkyl, (c) C2-C6 alkenyl, (d) C 3 -C6 cycloalkyl, (e) C3-C6 cycloalkenyl, and (f) fluorine, with the proviso that the total number of atoms, other than hydrogen, in each of R 8 , R 9 , and R 1 o, is 6 atoms or less. -568- WO 99/54299 PCT/US99/07945
2. The compound according to Claim 1 having the relative stereochemistry depicted by the formula: R 9 R Rio R 6 26 R2-X/ '...R "N R I R3 R 2 ,7, N R 1 R 7 \ wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
3. The compound according to Claim 1 having the relative stereochemistry depicted by formula : R 9 R 8 R 1 o R R2-X " N6 R4 R R5 R 3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 0 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same. -569- WO 99/54299 PCT/US99/07945
4. The compound according to Claim 1 wherein -X-R 2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2 -NH-SO 2 - or R 2 -SO 2 -NH wherein R 2 is 01-03 loweralkyl, halo C1-C3 loweralkyl, 02-03 alkenyl or halo C2-C3 alkenyl or -X-R 2 is Y1 -< Y y2 y2S N -/ -b) wherein Y 1 is -CH 2 -, -0-, -S- or -NH- and Y 2 is -C(=0)- or -C(Raa)( Rbb) - wherein Raa and Rbb are independently selected from the group consisting of hydrogen, C-C3 loweralkyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, aminomethyl, 1-aminoethyl, 2-aminoethyl, thiolmethyl, 1-thiolethyl, 2-thiolethyl, methoxymethyl, N-methylaminomethyl and methylthiomethyl; R 3 and R 4 are independently selected from hydrogen, heterocyclic and -Z-R 14 wherein Z and R 14 are as defined therein and wherein one of R 3 and R 4 is other than hydrogen; R 5 is hydrogen or loweralkyl; R 6 and R 7 are independently hydrogen or loweralkyl; R 8 and R 9 are independently hydrogen, fluoro or loweralkyl; R 10 is hydrogen, fluoro or loweralkyl; and Y is C2-C5 alkenyl, C2-C5 haloalkenyl, -C(=Q 2 )R 22 , -N(=Q 3 ), -N(O)=CHCH 3 , -NR 23 R 24 or a heterocyclic ring having from 3 to 6 ring atoms, wherein R 2 2 , R 23 , R 24 , Q 2 and Q3 are as defined therein. -570- WO 99/54299 PCT/US99/07945
5. The compound according to Claim 4 having the relative stereochemistry depicted by the formula: SR 9 R 8 6 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 o, X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
6. The compound according to Claim 4 having the relative stereochemistry depicted by formula : . R 9 R8 Rio R 2 1 R 2 -X N "R 1 R4R R" R3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
7. The compound according to Claim 1 wherein wherein -571- WO 99/54299 PCT/US99/07945 -X-R 2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2 -NH-SO 2 - or R 2 -SO 2 -NH wherein R 2 is 01-03 Ioweralkyl, halo C1-C3 loweralkyl, C2-C3 alkenyl or halo C2-C3 alkenyl or -X-R 2 iS 0 Y 1 _ 2 Y wherein Y 1 is -CH 2 - and Y 2 is -C(=O)- or -C(Raa)( Rbb) - wherein R aa and Rbb are independently selected from the group consisting of hydrogen, C1-C3 loweralkyl, hydroxymethyl, 1-hydroxyethyl and 2-hydroxyethyl; R 3 and R 4 are independently selected from hydrogen, heterocyclic and -Z-R 1 4 wherein Z and R 14 are as defined therein and wherein one of R 3 and R 4 is other than hydrogen; R 5 is hydrogen or loweralkyl; R 6 and R 7 are independently hydrogen or loweralkyl; R 8 and R 9 are independently hydrogen or loweralkyl; R 1 0 is hydrogen or loweralkyl; and Y is C2-C5 alkenyl, C2-05 haloalkenyl, -C(=Q 2 )R 22 , -N(=Q 3 ), -N(O)=CHCH 3 or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds, wherein R 22 , Q 2 and Q 3 are as defined therein. -572- WO 99/54299 PCT/US99/07945
8. The compound according to Claim 7 having the relative stereochemistry depicted by the formula: R 9 R8 Rio R 6 2-x R2- X,,, .. N "R 4 R 7 \R 5 R 3 R 3 wherein R', R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 0 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
9. The compound according to Claim 7 having the relative stereochemistry depicted by formula : . R 9 R8 Rio R 6 R 2 -X N "R 1 R4 R 7 5 R 3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
10. A compound according to Claim 1 wherein -573- WO 99/54299 PCT/US99/07945 -X-R 2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2 -NH-SO 2 - or R 2 -SO 2 -NH wherein R 2 is 01-03 loweralkyl, halo C1-C3 loweralkyl, 02-03 alkenyl or halo C1-C3 alkenyl or -X-R 2 is O Y 2 C Y wherein Y 1 is -OH 2 - and Y 2 is -C(=O)- or -C(Raa)( Rbb) - wherein Raa and Rbb are independently selected from the group consisting of hydrogen, C1-C3 loweralkyl, hydroxymethyl, 1-hydroxyethyl and 2-hydroxyethyl; R 3 and R 4 are independently selected from hydrogen, heterocyclic and -Z-R 14 wherein Z and R 14 are as defined therein and wherein one of R 3 and R 4 is other than hydrogen; R 5 is hydrogen or loweralkyl; R 6 and R 7 are independently hydrogen or loweralkyl; R 8 and R 9 are independently hydrogen or loweralkyl; R 10 is hydrogen or loweralkyl; and Y is 02-05 alkenyl, 02-C5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. -574- WO 99/54299 PCT/US99/07945
11. The compound according to Claim 10 having the relative stereochemistry depicted by the formula: R 9 R 8 Rio R6 R2 -X , N "R R4 R7 \ 5 R R R 3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
12. The compound according to Claim 10 having the relative stereochemistry depicted by formula : SR 9 R8 Rio R 6 R2-mX . Nl "*'",t R4 R Rs\ R 3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
13. A compound according to Claim 1 wherein R 1 is -CO 2 H; -575- WO 99/54299 PCT/US99/07945 -X-R 2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2 -NH-SO 2 - or R 2 -SO 2 -NH wherein R 2 is C 1 -0 3 loweralkyl or halo- C 1 -C 3 Ioweralkyl; R 3 and R 4 are independently selected from hydrogen, heterocyclic and -Z-R 14 wherein Z and R 14 are as defined therein and wherein one of R 3 and R 4 is other than hydrogen; R * 5 is hydrogen or loweralkyl; R 6 and R 7 are hydrogen independently hydrogen or loweralkyl; R 8 and R 9 are hydrogen independently hydrogen or loweralkyl; R 10 is hydrogen or loweralkyl; and Y is C2-C5 alkenyl, C 2 .- C 5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds.
14. The compound according to Claim 13 having the relative stereochemistry depicted by the formula: R 9 R 8 Rlo R6 R 2 - X .. N *R1 R4 R RS\ R R R 3 wherein R 1 , R 2 , R 3 , R 4 , Rs, R 6 , R 7 , R 8 , R 9 , R 1 0 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same. -576- WO 99/54299 PCT/US99/07945
15. The compound according to Claim 13 having the relative stereochemistry depicted by formula : SR 9 R8 6 Rio R R 2 -X N "R 1 R4 R 7 R 5 R 3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
16. The compound according to Claim 1 wherein R 1 is -CO 2 H; -X-R 2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2 -NH-SO 2 - or R 2 -SO 2 -NH wherein R 2 is 01-03 loweralkyl or halo- C 1 -C 3 loweralkyl; R 4 is hydrogen or loweralkyl and R 3 is heterocyclic or -Z-R 14 wherein Z and R 1 4 are as defined therein; R 5 is hydrogen; R 6 and R 7 are hydrogen; R 8 and R 9 are hydrogen; R 1 0 is hydrogen; and Y is C 2 -C 5 alkenyl, C 2 .C 5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. -577- WO 99/54299 PCT/US99/07945
17. The compound according to Claim 16 having the relative stereochemistry depicted by the formula: R 9 R8 Rio R 6 2 R X . . N R R R4 R 7 \R5 R 3 wherein R', R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 0 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
18. The compound according to Claim 16 having the relative stereochemistry depicted by formula : R 9 R8 Rio R 6 R 2 -X N "R RR R R 3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 0 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
19. The compound according to Claim 1 wherein R' is -CO 2 H; -578- -X-R 2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2 -NH-SO 2 - or R 2 -SO 2 -NH wherein R 2 is 01-C3 loweralkyl or halo C 1 -C 3 loweralkyl; R 4 is hydrogen or loweralkyl and R 3 is (a) heterocyclic, (b) alkyl, (d) cycloalkyl, (d) cycloalkylalkyl, (e) alkenyl, (f) alkynyl, (g) -C(=O)-R 14 (h) -C(R 3 7 a)(OR 37 c)-R1 4 or (i) -C(R 37 a)(R 37 b)-N(O)(R 3 7 c)R 14 wherein R 14 is (j) alkyl, (ii) cycloalkyl, (iii) cycloalkylalkyl, (iv) alkenyl, (v) haloalkyl, (vi) haloalkenyl, (vii) aryl, (viii) arylalkyl, (ix) heterocyclic, (x) (heterocyclic)alkyl, (xi) hydroxyalkyl, (xii) alkoxyalkyl, (xiii) cyanoalkyl, (xiv) (R 37 aO)-(O=)C-substituted alkyl or (xv) (R 37 aO) 2 -P(=O)-substituted alkyl; R 3 7 a and R 3 7 b are independently selected from the group consisting of (i) hydrogen, (ii) loweralkyl and (iii) loweralkenyl; and R 3 7 c is hydrogen, (ii) loweralkyl or (iii) loweralkenyl; R 5 is hydrogen; R 6 and R 7 are hydrogen; R 8 and R 9 are hydrogen; R 1 0 is hydrogen; and Y is C 2 -C 5 alkenyl, C2.-05 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. -579-
20. The compound according to Claim 19 having the relative stereochemistry depicted by the formula: R 9 8 Ri o ~R R2- X 1...N " R 1 R4 R 7 \5 R 3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
21. The compound according to Claim 19 having the relative stereochemistry depicted by formula : R 9 R 8 R 0 R 6 2-N "/R 1 R4 R R\ 5 R 3 wherein R 1 , R 2 , R 3 , R 4 , R s , R 6 , R 7 , R , R 9 , R 1 0 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
22. The compound according to Claim 1 wherein R 1 is -CO 2 H; -580- -X-R 2 is R 2 -C(=O)-NH-, R 2 -NH-C(=O)-, R 2 -NH-SO 2 - or R 2 -SO 2 -NH wherein R 2 is 01-03 loweralkyl or halo C1-C3 loweralkyl; R 4 is hydrogen and R 3 is (a) heterocyclic, (b) alkyl or (c) -C(R 37 a)(OR 37 c) R 14 wherein R 1 4 is (ii) alkyl, (ii) cycloalkyl, (iii) cycloalkylalkyl, (iv) alkenyl, (v) haloalkyl, (vi) haloalkenyl, (vii) aryl, (viii) arylalkyl, (ix) heterocyclic, (x) (heterocyclic)alkyl, (xi) hydroxyalkyl, (xii) alkoxyalkyl, (xiii) cyanoalkyl, (xiv) (R 3 7 aO)-(O=)C-substituted alkyl or (xv) (R 37 aO) 2 -P(=O)-substituted alkyl; R 3 7 a and R 3 7 b are independently selected from the group consisting of (i) hydrogen, (ii) loweralkyl and (iii) loweralkenyl; and R 3 7 c is hydrogen, (ii) C1-C3 loweralkyl or (iii) allyl; R 5 is hydrogen; R 6 and R 7 are hydrogen; R 8 and R 9 are hydrogen; R 10 is hydrogen; and Y is C2-C5 alkenyl, C2-C5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. -581-
23. The compound according to Claim 22 having the relative stereochemistry depicted by the formula: R 9 8 R R R10o- R6 R2-X,,,,.. N/ "",,,R' R4R 7 \R R3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 0 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
24. The compound according to Claim 22 having the relative stereochemistry depicted by formula : R 9 R Ro 10 R6 R 2 - xN R N "R 4RR 7 \R5 R3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , Ro 10 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
25. The compound according to Claim 1 wherein R 1 is-CO 2 H; -582- -X-R 2 is R 2 -C(=O)-NH- or R 2 -SO 2 -NH- wherein R 2 is C 1 -C 3 Ioweralkyl or halo C 1 -C 3 loweralkyl; R 4 is hydrogen and R 3 is (a) heterocyclic, (b) alkyl or (c) -C(R 37 a)(OR 37 c) R 14 wherein R 1 4 is (i) loweralkyl, (ii) loweralkenyl, (iii) hydroxy-substituted loweralkyl or (iv) alkoxy-substituted loweralkyl; R 3 7 a is (i) hydrogen, (ii) loweralkyl or (iii) loweralkenyl; and R 37 c is (i) hydrogen, (ii) C 1 -C 3 loweralkyl or (iii) allyl; R 5 is hydrogen; R 6 and R 7 are hydrogen; R 8 and R 9 are hydrogen; R 1 0 is hydrogen; and Y is C 2 -C 5 alkenyl, C 2 -C 5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds. -583-
26. The compound according to Claim 25 having the relative stereochemistry depicted by the formula: R 9 R 8 Rl o R 6 R2-X .'"/N R 1 R4 R 7R5 R3 wherein R', R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 0 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
27. The compound according to Claim 25 having the relative stereochemistry depicted by formula : R9 R R2-1 RN "R 4 R 7 \ 5 R 3 wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 o, X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
28. The compound according to Claim 1 wherein R 1 is -CO 2 H; -584- -X-R 2 is R 2 -C(=O)-NH- or R 2 -SO 2 -NH- wherein R 2 is Cl0-03 Ioweralkyl or halo C1-C3 loweralkyl; R 4 is hydrogen and R 3 is -C(R 37 a)(OR 3 7 c)-R1 4 wherein R 1 4 is loweralkyl or loweralkenyl; R 3 7 a is loweralkyl or loweralkenyl; and R 3 7 c is hydrogen, C-C3 loweralkyl or allyl; R 5 is hydrogen; R 6 and R 7 are hydrogen; R 8 and R 9 are hydrogen; R 1 0 is hydrogen; and Y is C2-C5 alkenyl, C2-C5 haloalkenyl or a heterocyclic ring having 5 ring atoms and also containing one or two double bonds.
29. The compound according to Claim 28 having the relative stereochemistry depicted by the formula: R 9 R 8 Y 5 R XR, 'R R 3 -585- wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
30. The compound according to Claim 29 having the relative stereochemistry depicted by formula : R9 8 R 4 0 R RR 2 RXN "Rz 1 R3 wherein R', R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1 0 , X and Y are as defined therein and wherein R 3 and R 4 are not both the same.
31. A compound selected from the group consisting of: (±)-(2R,3S,5R, 1'R)-2-(1-Acetamido-2-ethyl-2-hydroxy)butyl-3-(cis-propen- 1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-( 1 -Acetamido-2-hydroxy-2-methyl)pentyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-ethyl-2-hydroxy)pentyl-3-(cis-propen- 1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt; -586- (±)-(2R,3R,5R, 1'R,2'R)-2-(1-Acetamido-2,3-dihydroxy)propyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen-l yl)-pyrrolidine-5-carboxylic Acid ;. (-)-(2R,3S,5R, 1 'S)-2-(1-Acetamido-2-ethyl)butyl-3-(cis-propen.-1 -yl)-pyrrolidine-5 carboxylate Ammonium Salt; (±)-(2R,3S,5R, 1 'R,2'R)-2-(1 -Acetamido-2,3-dimethoxy)propyl-3-(cis-propen-l -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R, 3S, 5R, 1' R,2'S)-2-(1 -Acetamido-2-methoxy-2-vinyl)ethyl-3-(cis-propen -1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S, 5 R, 1 'S)-2-(1 -Acetamido-2-ethyl)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R,1'S,3'S)-2-(1 -Acetamido-2-(N-isopropyl-N-methylamino-N oxide))ethyl-3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1 'S,3'S)-2-(1-Acetamido-2-(N-ethyl-N-methylamino-N-oxide))ethyl 3-(cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R, 3S, 5R, 1'R,2'S)-2-(1 -Acetamido-2-methoxy)butyl-3-(cis-propen- 1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R,1'R,2'S)-2-(1 -Acetamido-2-methoxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid; (±)-(2R,3R,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy)butyl-3-(pyrazol-3-yl) pyrrolidine-5-carboxylic Acid; -587- (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Aceta mid o-2-hyd roxy) butyl-3-(cis-p ro pen- 1 -yI) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1 'R,2'R)-2-(1 -Acetamido-1 -(3,6-dihydro-2-H-pyran-2-yI))propyl-3 (cis-propen- 1 -yi)-pyrrolid ine-5-carboxylic Acid ; (±)-(2R, 3S,5R, 1 'R,2'S)-2-( 1 -Acetamido-2-methoxy-2-alyI)ethyl-3-(cis-.propen-1 yI)-pyrrolidine-5-carboxylic Acid; (±)-(2R,3S,5R, I 'R,2'S,3'S)-2-(1 -Aceta mid o-2-hyd roxy-3-methy1) pe nty 1-3- (cis propen-1 -yi)-pyrrolidine-5-carboxylic Acid ; (±)-(2 R, 3S,, 1 'R,2'S)-2-( 1 -Aceta mid o-2-meth oxy-4-vi nyl) buty- 3- (cisprope n-l yI)-pyrrolidine-5-carboxylic Acid ; (±)-(2R, 3S,5R, I 'R,2'S)-2-( 1 -Acetam id o-2- hyd roxy-3-cya no) propy 1-3-(cis-prope n . 1 -yI)-pyrrolidine-5-carboxylic Acid ; (±)-(2R, 3S ,5R, 1' R,2'S)-2-( 1 -Acetamido-1 -(3,6-d ihyd ro-2-H-pyran-2-yi))methyl-3 (cis-propen-1 -yI)-pyrrolid ine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamido-2,3-dimethoxy)propyl-3-(cs-propen-1 -yI) pyrrolidine-5-carboxylic Acid ; (±)-(2 R, 33,5R, 1' R,2'S)-2-( I -Aceta mid o-2- hyd roxymeth yl-2- hyd roxy) pe nty1- 3- (cis propen- 1 -yI)-pyrrolid ine-5-carboxylic Acid ; *(±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamido-2-ethoxy)pentyl-3-(cis-propen-1 -yl) pyrrolid ine-5-carboxylic Acid; (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-dimethyl)butyl-3-(cis-propen. 1 -yi)-pyrrolidine-5-carboxylic Acid -588- (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-ethoxy-3-vinyl)propyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R,1 'R,2'S)-2-(1-Acetamido-2-hydroxy-2-(propeny-2-yl))ethyl-3-(cis propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R,1 'R,2'S)-2-(1-Acetamido-2-hydroxy)hexyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid; (±)-(2R,3S, 5R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-(cis-propen-1-yl)-pyrrolidine 5-carboxylic Acid; (±)-(2R,3S,5R,1'R,2'S)-2-(1-Acetamido-2-hydroxy)butyl-3-vinyl-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy)pentyl-3-vinyl-pyrrolidine-5 carboxylic Acid; (±)-(2R,3S,5R,1'R,2'R)-2-(1-Acetamido-2-hydroxyethyl-2-hydroxy)pentyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'R)-2-(1 -Acetamido-2-hydroxy)butyl-3-vinyl-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'R)-2-(1 -Acetamido-2-methoxy)pentyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid; (±)-(2R,3S,5R, 1'R,2'R)-2-(1 -Acetamido-2-hydroxy)pentyl-3-vinyl-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R, 1'R)-2-(1 -Acetamido-2-hydroxy)ethyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid; -589- (±)-(2R,3S, 5 R, 1 'S)-2-(1-acetamido-3-methyl)butyl-3-(cis-2-chloro-vin-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R, 3 R,5R, 1 'S)-2-(1-acetamido-3-methyl)butyl-3-(pyrazol-3-yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3S,5R,1'S,3'R)-2-(1 -Acetamido-3-hydroxy)pentyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R, 3 R,5R,1'S)-2-(1-Acetamido-3-methyl)butyl-3-(thiazol-4-yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R,3R, 5 R,1'S)-1- t-Butoxycarbonyl-2-(1-acetamido-3-methyl)butyl-3-(thiazol 2-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R,1'S)-2-(1-Acetamido-3-methyl)butyl-3-vinyl-pyrrolidine-5-carboxylic Acid ; (±)-(2R, 3S,5R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-(2,2-difluoro-vin- 1 -yl) pyrrolidine-5-carboxylic Acid; (±)-(2R, 3R, 5R, 1 'S)-2-(1-acetamido-3-methyl)butyl-3-(pyrazol-3-yl)-pyrrolidine-5 carboxylic Acid; (±)-(2R, 3R,5R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-(isoxazol-3-yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R, 3R, 5R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-(isoxazol-5-yl)-pyrrolidine-5 carboxylic Acid ; (±)-(2R, 3R, 5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(imidazol-2-yl)-pyrrolidine-5 Carboxylic Acid; -590- (±)-(2R,3R,5R, 1 'S)-2-(1 -Acetamido-3-methyl)butyl-3-(imidazol-4-yl)-pyrrolid ine-5 carboxylic Acid ; and (±)-(2S,3R,5R, 1 'S)-2-(1 -acetamido-3-methyl)butyl-3-amino-pyrrolidine-5 carboxylic Acid; or a pharmaceutically acceptable salt, ester or prodrug thereof.
32. A compound selected from the group consisting of: (±)-(2R,3S,5R, 1'R)-2-(1 -Acetamido-2-ethyl-2-hydroxy)butyl-3-(cis-propen- 1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-2-methyl)pentyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-( 1 -Acetamido-2-ethyl-2-hydroxy)pentyl-3-(cis-propen- 1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy)pentyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid Trifluoroacetic Acid Salt; (±)-(2R,3R,5R, 1'R,2'R)-2-(1 -Acetamido-2,3-dihydroxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxy-4-vinyl)butyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ;. (-)-(2R, 3S, 5R, 1 'S)-2-(1-Acetamido-2-ethyl)butyl-3-(cis-propen-1-yl)-pyrrolidine-5 carboxylate Ammonium Salt; (±)-(2R,3S,5R, 1'R,2'R)-2-(1 -Acetamido-2,3-dimethoxy)propyl-3-(cis-propen- 1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-methoxy-2-vinyl)ethyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; -591- (±)-(2R,3S,5R, 1 'S)-2-(1 -Aceta mid o-2-ethyl) butyl-3-(cis-p rope n-l -yi)-pyrrolidine-5 carboxylic Acid (±)-(2R,3S,5R, 1 'S,3'S)-2-(1 -Acetamido-2-(N-isopropyl-N-methylamino-N oxide))ethyl-3-(cis-propen- 1 -yI)-pyrrolidine-5-carboxylic Acid ; (±)-(2 R, 3S.5 R, 1'S, 3'S)-2-( 1 -Acetamido-2-(N-ethyl-N-methylamino-N-oxide))ethyl 3-(cis-propen-1 -yi)-pyrrolid ine-5-carboxylic Acid ; (±)-(2R, 3S ,5R, 1 'R,2'S)-2-( 1 -Acetamid o-2-methoxy) butyl-3-(cis-p rope n- 1 -yI) pyrrolidine-5-carboxylic Acid ; (±)-(2 R,3S ,5R, 1' R,2'S)-2-( 1 -Aceta mid o-2-m eth oxy) pe ntyi-3- (cis-p ropen- 1l-yi) pyrrolid ine-5-carboxylic Acid ; (±)-(2 R, 3R, 5R, 1' R,2'S)-2-( 1 -Aceta mid o-2-hyd roxy) butyl-3-(pyrazol-3-yi) pyrro lid ine-5-carboxylic Acid ; (±)-(2R, 3S, SR, 1 'R,2'S)-2-( 1 -Acetamid o-2-hyd roxy)butyl-3-(cis-p rope n- 1 -yI) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1 'R,2'R)-2-(1 -Acetamido-1 -(3,6-dihydro-2-H-pyran-2-yl))propyl-3 (cis-propen-1 -yI)-pyrrolid ine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamido-2-methoxy-2-ally)ethyl-3-(cis-propen-1 yI)-pyrrolidine-5-carboxylic Acid; (±)-(2R,3S,5R, 1'R,2'S,3'S)-2-(1 -Acetamido-2-hydroxy-3-methyl)pentyl-3-(cis propen-1 -yI)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamido-2-methoxy-4-vinyl)butyl-3-(cis-propenl1 yI)-pyrrolidine-5-carboxylic Acid; (±)-(2R,3S,5R, 1 'R,2'S)-2-(1 -Acetamid o-2- hyd roxy-3-cya no) propyl-3-(cis-p rope n 1-yI)-pyrrolidine-5-carboxylic Acid -592- (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-1 -(3,6-dihydro-2-H-pyran-2-yl))methyl-3 (cis-propen-1 -yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2,3-dimethoxy)propyl-3-(cis-propen-1 -yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1-Acetamido-2-hydroxymethyl-2-hydroxy)pentyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-ethoxy)pentyl-3-(cis-propen-1-yl) pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-3-dimethyl)butyl-3-(cis-propen 1-yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-ethoxy-3-vinyl)propyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; (±)-(2R,3S,5R, 1'R,2'S)-2-(1 -Acetamido-2-hydroxy-2-(propeny-2-yl))ethyl-3-(cis propen-1-yl)-pyrrolidine-5-carboxylic Acid ; and (±)-(2R,3S,5R,1'R,2'S)-2-(1 -Acetamido-2-hydroxy)hexyl-3-(cis-propen-1 yl)-pyrrolidine-5-carboxylic Acid ; or a pharmaceutically acceptable salt, ester or prodrug thereof.
33. A pharmaceutical composition for inhibiting influenza neuraminidase comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of Claim 1.
34. A pharmaceutical composition for treating an influenza infection comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of Claim 1. -593-
35. A pharmaceutical composition for preventing an influenza infection comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of Claim 1.
36. A pharmaceutical composition for inhibiting influenza neuraminidase comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of Claim 31.
37. A pharmaceutical composition for treating an influenza infection comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of Claim 31.
38. A pharmaceutical composition for preventing an influenza infection comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of Claim 31.
39. A method for inhibiting neuraminidase from a disease-causing microorganism comprising administering to a human or other mammal in need thereof, a therapeutically effective amount of a compound of Claim 1.
40. The method of Claim 39 wherein the disease-causing microorganism is a virus.
41. The method of Claim 40 wherein the virus is influenza virus.
42. A method for treating a disease caused by a microorganism which has a neuraminidase, comprising administering to a human or other mammal in need thereof, a therapeutically effective amount of a compound of Claim 1.
43. The method of Claim 42 wherein the disease-causing microorganism is a virus.
44. The method of Claim 43 wherein the virus is influenza virus. -594-
45. A method for preventing a disease caused by a microorganism which has a neuraminidase, comprising administering to a human or other mammal in need thereof, a therapeutically effective amount of a compound of Claim 1.
46. The method of Claim 45 wherein the disease-causing microorganism is a virus.
47. The method of Claim 46 wherein the virus is influenza virus.
48. A method for inhibiting neuraminidase from a disease-causing microorganism comprising administering to a human or other mammal in need thereof, a therapeutically effective amount of a compound of Claim 31.
49. The method of Claim 48 wherein the disease-causing microorganism is a virus.
50. The method of Claim 49 wherein the virus is influenza virus.
51. A method for treating a disease caused by a microorganism which has a neuraminidase, comprising administering to a human or other mammal in need thereof, a therapeutically effective amount of a compound of Claim 31.
52. The method of Claim 51 wherein the disease-causing microorganism is a virus.
53. The method of Claim 52 wherein the virus is influenza virus.
54. A method for preventing a disease caused by a microorganism which has a neuraminidase, comprising administering to a human or other mammal in need thereof, a therapeutically effective amount of a compound of Claim 31.
55. The method of Claim 54 wherein the disease-causing microorganism is a virus. -595-
56. The method of Claim 55 wherein the virus is influenza virus. -596-
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US6522598A | 1998-04-23 | 1998-04-23 | |
| US09065225 | 1998-04-23 | ||
| PCT/US1999/007945 WO1999054299A1 (en) | 1998-04-23 | 1999-04-12 | Pyrrolidines as inhibitors of neuraminidases |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| AU3554599A true AU3554599A (en) | 1999-11-08 |
Family
ID=22061195
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| AU35545/99A Abandoned AU3554599A (en) | 1998-04-23 | 1999-04-12 | Pyrrolidines as inhibitors of neuraminidases |
Country Status (17)
| Country | Link |
|---|---|
| EP (1) | EP1315698A1 (en) |
| JP (1) | JP2002512224A (en) |
| CN (1) | CN1328546A (en) |
| AR (1) | AR018196A1 (en) |
| AU (1) | AU3554599A (en) |
| BG (1) | BG104962A (en) |
| BR (1) | BR9909870A (en) |
| CA (1) | CA2329422A1 (en) |
| CO (1) | CO5011122A1 (en) |
| HU (1) | HUP0101224A3 (en) |
| IL (1) | IL138601A0 (en) |
| NO (1) | NO20005301L (en) |
| PL (1) | PL343678A1 (en) |
| SK (1) | SK15092000A3 (en) |
| TR (1) | TR200003065T2 (en) |
| WO (1) | WO1999054299A1 (en) |
| ZA (1) | ZA200005238B (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ATE322163T1 (en) | 1996-06-14 | 2006-04-15 | Biocryst Pharm Inc | SUBSTITUTED CYCLOPENTAN DERIVATIVES USEFUL AS NEURAMIDASE INHIBITORS |
| SI1040094T1 (en) | 1997-12-17 | 2009-10-31 | Biocryst Pharm Inc | Substituted cyclopentane and cyclopentene compounds useful as neuraminidase inhibitors |
| US6455571B1 (en) * | 1998-04-23 | 2002-09-24 | Abbott Laboratories | Inhibitors of neuraminidases |
| WO2000028328A1 (en) | 1998-11-05 | 2000-05-18 | Biocryst Pharmaceuticals, Inc. | New cyclopentane and cyclopentene compounds and use for detecting influenza virus |
| WO2001080892A1 (en) * | 2000-04-25 | 2001-11-01 | Sankyo Company, Limited | Preventives for influenza |
| US6518299B1 (en) | 2000-10-20 | 2003-02-11 | Biocryst Pharmaceuticals, Inc. | Substituted pyrrolidine compounds useful as neuraminidase inhibitors |
| WO2002081441A1 (en) * | 2001-04-03 | 2002-10-17 | Abbott Laboratories | Process for the preparation of substituted pyrrolidine neuraminidase inhibitors |
| JP2005511573A (en) * | 2001-11-02 | 2005-04-28 | グラクソ グループ リミテッド | 4- (5-membered) -heteroarylacylpyrrolidine derivatives as HCV inhibitors |
| WO2003037895A1 (en) * | 2001-11-02 | 2003-05-08 | Glaxo Group Limited | 4-(6-membered)-heteroaryl acyl pyrrolidine derivatives as hcv inhibitors |
| EP1554274B1 (en) * | 2002-10-24 | 2008-12-24 | Glaxo Group Limited | 1-acyl-pyrrolidine derivatives for the treatment of viral infections |
| CN104402754B (en) * | 2014-11-25 | 2016-03-02 | 广东东阳光药业有限公司 | As the compound of neuraminidase inhibitor and the application in medicine thereof |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4035961A1 (en) * | 1990-11-02 | 1992-05-07 | Thomae Gmbh Dr K | CYCLIC IMINODERIVATES, MEDICAMENTS CONTAINING SUCH COMPOUNDS AND METHOD FOR THE PRODUCTION THEREOF |
| DE10299052I1 (en) * | 1995-02-27 | 2003-05-22 | Gilead Sciences Inc | New selective inhibitors of viral or bacterial neuraminidases |
| ATE322163T1 (en) * | 1996-06-14 | 2006-04-15 | Biocryst Pharm Inc | SUBSTITUTED CYCLOPENTAN DERIVATIVES USEFUL AS NEURAMIDASE INHIBITORS |
| ZA988469B (en) * | 1997-09-17 | 1999-03-17 | Biocryst Pharm Inc | Pyrrolidin-2-one compounds and their use as neuraminidase inhibitors |
-
1999
- 1999-04-12 SK SK1509-2000A patent/SK15092000A3/en unknown
- 1999-04-12 IL IL13860199A patent/IL138601A0/en unknown
- 1999-04-12 CA CA002329422A patent/CA2329422A1/en not_active Abandoned
- 1999-04-12 CN CN99807610A patent/CN1328546A/en active Pending
- 1999-04-12 BR BR9909870-9A patent/BR9909870A/en not_active IP Right Cessation
- 1999-04-12 HU HU0101224A patent/HUP0101224A3/en unknown
- 1999-04-12 AU AU35545/99A patent/AU3554599A/en not_active Abandoned
- 1999-04-12 JP JP2000544640A patent/JP2002512224A/en not_active Withdrawn
- 1999-04-12 PL PL99343678A patent/PL343678A1/en unknown
- 1999-04-12 EP EP99917414A patent/EP1315698A1/en not_active Withdrawn
- 1999-04-12 WO PCT/US1999/007945 patent/WO1999054299A1/en not_active Application Discontinuation
- 1999-04-12 TR TR2000/03065T patent/TR200003065T2/en unknown
- 1999-04-21 CO CO99024047A patent/CO5011122A1/en unknown
- 1999-04-23 AR ARP990101905A patent/AR018196A1/en not_active Application Discontinuation
-
2000
- 2000-09-28 ZA ZA200005238A patent/ZA200005238B/en unknown
- 2000-10-20 NO NO20005301A patent/NO20005301L/en not_active Application Discontinuation
- 2000-11-17 BG BG104962A patent/BG104962A/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| HUP0101224A2 (en) | 2001-08-28 |
| TR200003065T2 (en) | 2001-02-21 |
| IL138601A0 (en) | 2001-10-31 |
| BG104962A (en) | 2001-07-31 |
| BR9909870A (en) | 2000-12-19 |
| CO5011122A1 (en) | 2001-02-28 |
| SK15092000A3 (en) | 2001-05-10 |
| AR018196A1 (en) | 2001-10-31 |
| CA2329422A1 (en) | 1999-10-28 |
| ZA200005238B (en) | 2001-12-04 |
| EP1315698A1 (en) | 2003-06-04 |
| NO20005301D0 (en) | 2000-10-20 |
| PL343678A1 (en) | 2001-08-27 |
| NO20005301L (en) | 2000-12-08 |
| WO1999054299A1 (en) | 1999-10-28 |
| CN1328546A (en) | 2001-12-26 |
| HUP0101224A3 (en) | 2002-12-28 |
| JP2002512224A (en) | 2002-04-23 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MK5 | Application lapsed section 142(2)(e) - patent request and compl. specification not accepted |