HK1173438A

HK1173438A - Processes and intermediates

Info

Publication number: HK1173438A
Application number: HK13100325.1A
Authority: HK
Inventors: Gerald J. Tanoury; Minzhang Chen; John E. Cochran; Adam Looker; Valdas Jurkauskas
Original assignee: Vertex Pharmaceuticals Incorporated
Priority date: 2009-04-27
Filing date: 2010-04-27
Publication date: 2013-05-16

Description

Processes and intermediates

Cross-referencing

This application is PCT at 12/430,207 filed on 27/4/2009, which is a continuation-in-part application of us serial No. 11/506,550 filed on 18/8/2006, which claims priority from us codex 35, chapter 119(e) of the united states code, us serial No. 60/709,964 filed on 19/8/2005 and 60/810,042 filed on 1/6/2006, each of which is incorporated herein in its entirety.

Technical Field

The present invention relates to processes and intermediates for the preparation of protease inhibitors, particularly serine protease inhibitors.

Background

Hepatitis c virus ("HCV") infection is a human medical problem of concern. HCV is considered the causative agent of most cases of non-A, non-B Hepatitis, with an estimated global human seroprevalence of 3% (A. Alberti et al, "Natural History of Hepatitis C (Natural History of Hepatitis C)," J. hepatology, 31(suppl.1), pp.17-24 (1999)). There may be approximately 4 million people infected in the united states alone. (M.J.Alter et al, "The Epidemiology of viral Hepatitis in The United States", "gastroenterol.Clin.North Am., 23, 437-455 (1994); M.J.Alter" Hepatitis C Virus Infection in The United States "," J.Heatology, 31(Suppl.1), pp.88-91 (1999)).

Only about 20% of infected individuals develop acute clinical hepatitis after the first exposure to HCV, while others appear to resolve the infection spontaneously. However, in almost 70% of cases, virus establishment can persist for decades as a chronic infection. (S.Iward, "The Natural horse of Chronic Hepatitis (Natural process of chronic Hepatitis)," FEMS Microbiology Reviews, 14, 201 pages 204 (1994); D.Lavanchy, "Global scientific and Control of Hepatitis C)," J.visual Hepatitis, 6, 35-47 (1999)). Prolonged chronic infection can lead to recurrent and progressive worsening of liver inflammation, which often leads to more severe diseases such as cirrhosis and hepatocellular carcinoma. (M.C.Kew, "Hepatitis Cand Hepatocellular Carcinoma (Hepatitis C and Hepatocellular Carcinoma)", FEMSMIC microbiology Reviews, 14, 211-. Unfortunately, there is no widely effective treatment for chronic HCV to progressively debilitate.

Compounds described as protease inhibitors, in particular serine protease inhibitors, useful for the treatment of HCV infections are disclosed in WO 02/18369. Also disclosed in this publication are processes and intermediates for preparing these compounds which result in racemization of certain stereogenic carbon centers. See, e.g., pages 223-22. However, there is still a need for economical processes for preparing these compounds.

Summary of The Invention

In one aspect, the present invention provides processes and intermediates for preparing bicyclic pyrrolidine derivatives of formula 1, which are useful for preparing protease inhibitors.

In formula 1, R₃Are acid protecting groups which can be removed under acidic, basic or hydrogenation conditions. Under acidic conditions, R₃Is, for example, tert-butyl; under alkaline conditions, R₃Is for example methyl or ethyl; under hydrogenation conditions, R₃Is, for example, benzyl.

Another aspect of the invention includes processes and intermediates for preparing compounds of formula 2, which are also useful for preparing protease inhibitors.

In the formula 2, the first and second groups,

R₄is H, optionally substituted aliphatic, optionally substituted cycloaliphatic, optionally substituted aryl, optionally substituted aralkyl or optionally substituted heteroaralkyl;

R′₄is H, optionally substituted aliphatic, optionally substituted aryl, optionally substituted aralkyl or optionally substituted heteroaralkyl; and

R′₅is optionally substituted aliphatic, optionally substituted cycloaliphatic, optionally substituted aryl, optionally substituted aralkyl or optionally substituted heteroaralkyl; or

R′₄And R'₅Together with the atoms to which they are attached may form a 3-to 7-membered optionally substituted cycloaliphatic ring.

The processes and intermediates described herein are also useful in processes for preparing the protease inhibiting compounds of formula 3 shown below.

With reference to the formula 3, the following description is given,

R₁is RW-, P₂-、P₃-L₂-P₂-or P₄-L₃-P₃-L₂-P₂-；

P₂-is

P₃-L₂-P₂Is that

P₄-L₃-P₃-L₂-P₂Is that

W is a bond, -CO-, -O-CO-, -NR^X-、-NR^X-CO-, -O-or-S-;

t is-C (O) -, -O-C (O) -, -NHC (O) -, -C (O) -or-SO₂-；

R is H, optionally substituted aliphatic, optionally substituted cycloaliphatic, optionally substituted heterocycloaliphatic, optionally substituted aryl, or optionally substituted heteroaryl;

R₅is H, aliphatic, cycloaliphatic, heterocycloaliphatic, aryl or heteroaryl; each of which is optionally substituted, in addition to H, with one or more substituents each independently selected from Group J, wherein Group J comprises halo, cycloaliphatic, aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl, nitro, cyano, amido, amino, sulfonyl, sulfinyl, thio (sulfenyl), sulfone, ureido, thioureido, sulfamoyl, sulfonamide, oxo, carboxyl, carbamoyl, cycloaliphatic oxy, heterocycloaliphatic oxy, aryloxy, heteroaryloxy, aralkoxy, heteroarylalkoxy, alkoxycarbonyl, alkylcarbonyloxy, and hydroxy;

R₆is optionally substituted aliphatic, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted phenyl; or R₅And R₆Together with the atoms to which they are attached, may form a 5-to 7-membered optionally substituted monocyclic heterocycle, or a 6-to 12-membered optionally substituted bicyclic heterocycle, wherein each heterocycle optionally contains an additional moiety selected from-O-, -S-, or-NR^X-a heteroatom of (a);

R₇and R₇' are each independently H, optionally substituted aliphatic, optionally substituted heteroalkyl, optionally substituted heteroaryl, or optionally substituted phenyl; or R₇And R₇', together with the atoms to which they are attached, may form a 3-to 7-membered cycloaliphatic or heterocycloaliphatic ring; or

R₇And R₆Together with the atoms to which they are attached, may form a 5-to 7-membered optionally substituted monocyclic heterocycle, a 5-to 7-membered optionally substituted monocyclic aryl, a 6-to 12-membered optionally substituted bicyclic heterocycle or a 6-to 12-membered optionally substituted bicyclic aryl, wherein each heterocycle or aryl ring optionally contains an additional moiety selected from-O-, -S-or-NR^X-a heteroatom of (a); or

When R is₅And R₆When they form a ring together with the atom to which they are attached, R₇And by R₅And R₆The ring system formed may form an 8-to 14-membered optionally substituted bicyclic fused ring system, wherein the bicyclic fused ring system may be further fused with an optionally substituted phenyl to form an optionally substituted 10-to 16-membered tricyclic fused ring system;

R₈is H or a protecting group;

R^Xis H, aliphatic, cycloaliphatic, (cycloaliphatic) aliphatic, aryl, araliphatic, heterocycloaliphatic, (heterocycloaliphatic) aliphatic, heteroaryl, carboxy, thio, sulfinyl, sulfonyl, (aliphatic) carbonyl, (cycloaliphatic) carbonyl, ((cycloaliphatic) aliphatic) carbonyl, arylcarbonyl, (araliphatic) carbonyl, (heterocycloaliphatic) carbonyl, ((heterocycloaliphatic) aliphatic) carbonyl, (heteroaryl) carbonyl, or (heteroarylaliphatic) carbonyl;

R₂is- (NH-CR)₄′R₅′-C(O)-C(O))-NHR₄Or- (NH-CR)₄′R₅′-CH(OH)-C(O))-NHR₄；

R₄Is H, optionally substituted aliphatic, optionally substituted cycloaliphatic, optionally substituted heterocycloaliphatic, optionally substituted aryl, optionally substituted aralkyl or optionally substituted heteroaralkyl; and

R′₄and R'₅Each independently is H, optionally substituted aliphatic, optionally substituted cycloaliphatic, optionally substituted heterocycloaliphatic, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaralkyl or optionally substituted heteroaralkyl; or R₄' and R₅', together with the atoms to which they are attached, may form a 3-to 7-membered optionally substituted cycloaliphatic ring.

In some embodiments, the process for preparing the compound of formula 3 comprises the step of carboxylation of azabicyclooctane of formula 6,

6

wherein R' is C_1-5Alkyl to give cis-and trans-octahydrocyclopenta [ c ] s of formula 7]Racemic mixtures of pyrrole-1-carboxylic acid.

In some embodiments, P₂、P₃And P₄Each independently is a bond, H, optionally substituted aliphatic, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkoxy, optionally substituted alkylthio, optionally substituted aralkoxy, optionally substituted aralkylthio, optionally substituted mono-or dialkylamino, optionally substituted mono-or diarylamino, or optionally substituted mono-or diheteroarylamino.

In some embodiments, L is₂And L₃Each independently is a bond, -C (O) -or-SO₂-。

In some embodiments, R₅Is C_1-6Alkyl radical, C_3-10Cycloalkyl radical, C_3-10cycloalkyl-C_1-12Alkyl radical, C_6-10Aryl radical, C_6-10aryl-C_1-6Alkyl radical, C_3-10Heterocyclic group, C_6-10heterocyclyl-C_1-6Alkyl radical, C_5-10Heteroaryl or C_5-10heteroaryl-C_1-6An alkyl group; each of which is optionally substituted with 1-3 substituents each independently selected from Group J; and at R₅Up to 3 aliphatic carbon atoms in the group consisting of O, NH, S, SO and SO₂Independently replaced in a chemically stable arrangement.

In some further embodiments, R₅Is that

In some embodiments, R₇' is H; r₇Is C_1-6Alkyl radical, C_3-10Cycloalkyl radical, C_3-10cycloalkyl-C_1-12Alkyl radical, C_6-10Aryl radical, C_6-10aryl-C_1-6Alkyl radical, C_3-10Heterocyclic group, C_6-10heterocyclyl-C_1-6Alkyl radical, C_5-10Heteroaryl or C_5-10heteroaryl-C_1-6An alkyl group; r₁Is optionally substituted with 1-3 substituents each independently selected from Group J; and at R₁Up to 3 aliphatic carbon atoms in the group consisting of O, NH, S, SO and SO₂Are replaced in a chemically stable arrangement.

In some further embodiments, R₇Is that

In still some further embodiments, R₇And R₇', together with the atoms to which they are attached, form

In some embodiments, R is C_6-10Aryl radical, C_6-10aryl-C_1-12Aliphatic, C_3-10Cycloalkyl radical, C_3-10Cycloalkenyl radical, C_3-10cycloalkyl-C_1-12Aliphatic, C_3-10Cycloalkenyl radical-C_1-12Aliphatic, C_3-10Heterocyclic group, C_3-10heterocyclyl-C_1-12Aliphatic, C_5-10Heteroaryl or C_5-10heteroaryl-C_1-12Aliphatic; each of which is optionally substituted with 1-3 substituents each independently selected from GroupJ.

In some further embodiments, R is

In still some further embodiments, R is

And R is₁₀Is H, C_1-12Aliphatic, C_6-10Aryl radical, C_6-10aryl-C_1-12Aliphatic, C_3-10Cycloalkyl radical, C_3-10Cycloalkenyl radical, C_3-10cycloalkyl-C_1-12Aliphatic, C_3-10Cycloalkenyl radical-C_1-12Aliphatic, C_3-10Heterocyclic group, C_3-10heterocyclyl-C_1-12Aliphatic, C_5-10Heteroaryl or C_5-10heteroaryl-C_1-12Aliphatic.

In still some further embodiments, R is

In some further embodiments, R is

In some embodiments, the carboxylation step of the process for preparing the compound of formula 3 comprises forming the 2-anion of the compound of formula 6 in the presence of a complexing agent,

the 2-anion is then treated with carbon dioxide to produce a racemic mixture of the formula 7 trans-/cis-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid

In some further embodiments, the 2-anion of the compound of formula 6 is prepared by treating the compound of formula 6 with a strong lithium base in the presence of a complexing agent and an aprotic solvent.

In still some further embodiments, the base used to prepare the 2-anion is sec-butyllithium.

In still further embodiments, the complexing agent used to prepare the 2-anion is tetramethylethylenediamine, tetraethylethylenediamine, tetramethyl-1, 2-cyclohexyldiamine, sparteine, or 3, 7-bis (C)_1-6Alkyl) -3, 7-diazabicyclo [3.3.1]Nonanes, e.g. 3, 7-di (n-propyl) -3, 7-diazabicyclo [3.3.1]Nonane.

In still further embodimentsIn one embodiment, the complexing agent is tetramethylethylenediamine, tetraethylethylenediamine, tetramethyl-1, 2-cyclohexyldiamine, or 3, 7-bis (C)_1-6Alkyl) -3, 7-diazabicyclo [3.3.1]Nonane.

In still further embodiments, the complexing agent is D-sparteine.

In some embodiments, the trans-/cis-ratio in the racemic mixture of the compound of formula 7 is 1: 2.

In some embodiments, the trans-/cis-ratio in the racemic mixture of the compound of formula 7 is 40: 60.

In still further embodiments, the trans-/cis-ratio in the racemic mixture of the compound of formula 7 is 1: 1.

In still further embodiments, the trans-/cis-ratio is 60: 40.

In still further embodiments, the trans-/cis-ratio is 80: 20.

In still further embodiments, the trans-/cis-ratio is 90: 10.

In still further embodiments, the trans-/cis-ratio is greater than 98: 2.

In some other embodiments, the method of preparing the compound of formula 3 further comprises balancing the compound of formula 7 in the presence of a suitable base

To produce a trans-predominantly cis-racemic acid of formula 8

Wherein the trans-/cis-ratio is greater than 80: 20.

In some other embodiments, the process for preparing the compound of formula 3 further comprises balancing a trans-/cis-mixture of the compound of formula 7 in the presence of a suitable base to produce a trans-dominant-cis racemic acid of formula 8, wherein the trans-/cis-ratio is greater than 90: 10.

In some other embodiments, the process for preparing the compound of formula 3 further comprises balancing the trans-/cis-mixture of formula 7 in the presence of a suitable base to produce a trans-dominant-cis racemic acid of formula 8, wherein the trans-/cis-ratio is greater than 98: 2.

In some further embodiments, the base used to balance the 7 trans-/cis-mixture is lithium hexamethyldisilazide, lithium diisopropylamide, or lithium 2, 2, 6, 6-tetramethylpiperidine.

In some further embodiments, the base is lithium hexamethyldisilazide.

In some further embodiments, the base is sec-butyllithium and the complexing agent is 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] nonane to provide a racemic mixture of trans-/cis-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid of formula 7, wherein the trans-/cis-ratio is greater than 90: 10.

In some further embodiments, the trans-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid is trans-N-tert-butoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid.

In some other embodiments, the process for preparing the compound of formula 3 further comprises isolating racemic trans-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid to produce (1S, 2S, 3R) trans-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid.

In some further embodiments, the separation of the racemic mixture of compounds comprises the steps of i) forming a salt with an optically active base; and ii) crystallizing the salt formed by step i) to provide the optically active salt of formula 9.

In some further embodiments, the optically active base used to separate the racemic mixture of compounds is (R) alpha-aminoethylbenzene.

In some further embodiments, the optically active base is (S)1, 2, 3, 4-tetrahydro-1-naphthylamine.

In some further embodiments, the process for preparing the compound of formula 3 further comprises reacting a carboxylic acid of formula 9 with a compound containing R₃Esterification of a compound of the group; and a step of removing the-COOR' protecting group to give a compound of formula 1,

wherein R is₃Is an optionally substituted alkyl or aralkyl group.

In still some further embodiments, R₃Is a tert-butyl group.

In some embodiments, the method of preparing a compound of formula 3 further comprises reacting an amino-ester of formula 1 with R₁COOH is reacted in the presence of a coupling reagent to yield the compound of formula 1 a.

In some embodiments, the amino-ester of formula 1 is substituted with R₁The reaction between COOH may further be carried out in addition to the coupling agentIn the presence of histamine, glycine or lysine.

In some further embodiments, R₁Is P₂-。

In some further embodiments, R₁Is P₃-L₂-P₂-。

In some further embodiments, R₁Is P₄-L₃-P₃-L₂-P₂-。

In some further embodiments, R₁Is RW-.

In some embodiments, the method of preparing a compound of formula 3 further comprises the steps of: hydrolyzing an ester of the compound of formula 1 a; to provide a carboxylic acid and reacting the carboxylic acid thus obtained with a compound containing R₂Reacting the compound of the group in the presence of a coupling reagent to produce a compound of formula 3, wherein R₂Is- (NH-CR)₄′R₅′-CH(OH)C(O))-NHR₄。

In some further embodiments, R₄Is H, optionally substituted aliphatic, optionally substituted cycloaliphatic, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aralkyl or optionally substituted heteroaralkyl;

R₄' is H, optionally substituted aliphatic, optionally substituted aryl, optionally substituted aralkyl or optionally substituted heteroaralkyl; and

R₅' is H, optionally substituted aliphatic, optionally substituted cycloaliphatic, optionally substituted aryl, optionally substituted aralkyl or optionally substituted heteroaralkyl; or

R₄' and R₅', together with the atoms to which they are attached, form a 3-to 7-membered optionally substituted cycloaliphatic ring.

In some further embodiments, R₂Is that

The invention further relates to a process for the preparation of a compound of formula 4

In some embodiments, the method of preparing the compound of formula 4 comprises the steps of:

i) providing N-alkoxycarbonyl-3-azabicyclo [3.3.0] octane;

ii) a 2-anion which forms N-alkoxycarbonyl-3-azabicyclo [3.3.0] octane in the presence of a chelating agent;

iii) treating the anion of step ii) with carbon dioxide to produce a cis-/trans-mixture of N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid;

iv) treating the mixture of step iii) with a strong base to produce substantially pure trans-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid;

v) forming a salt of a carboxylic acid with an optically active amine;

vi) crystallizing the salt;

vii) esterifying the salt provided in step vi);

viii) removal of the N-alkoxycarbonyl group to yield (1S, 3aR, 6aS) -tert-butyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid tert-butyl ester;

ix) contacting the bicyclic ring of step viii) with a protected amino acid of formula 26,

wherein Z is an amine protecting group, in the presence of a coupling reagent to produce an amide-ester of formula 27;

x) removing the protecting group Z from the amide-ester of step ix) to yield an amino compound of formula 28;

xi) reacting an amino compound of formula 28 with a protected amino acid of formula 29

Reacting in the presence of a coupling reagent to produce the tripeptide of formula 30;

xii) removing the protecting group Z from the tripeptide of formula 30 to yield a free amino-tripeptide of formula 31;

xiii) reacting the amino-tripeptide of formula 31 with pyrazine-2-carboxylic acid in the presence of a coupling reagent to yield an amide-tripeptide ester of formula 33;

xiv) hydrolyzing the ester of the amide-tripeptide ester of formula 33 to yield the amide-tripeptide acid of formula 34;

xv) reacting an amide-tripeptide acid of formula 34 with an aminohydroxy-amide of formula 18

Reacting in the presence of a coupling reagent to produce a hydroxy-tetrapeptide of formula 35; and

xvi) oxidizing the hydroxy group of formula 35 to produce the compound of formula 4.

In some embodiments, the oxidizing agent used in step xvi) described above is sodium hypochlorite, and the oxidation is carried out in the presence of 2, 2, 6, 6-tetramethylpiperidinyloxy radical (TEMPO).

In some other embodiments, the oxidizing agent used in step xvi) described above is 1, 1-dihydro-1, 1, 1-triacetoxy-1, 2-benziodoxazol-3 (1H) -one (benzodioxool-3 (1H) -one).

In some further embodiments, the method further comprises dissolving the compound of formula 4 in an organic solvent to obtain a solution thereof, and then adding an acid to the solution. Suitable organic solvents may be any solvent in which the compound of formula 4 is dissolved, such as dichloromethane. The acid may be any acid, inorganic or organic, such as acetic acid or propionic acid.

In still some further embodiments, the method further comprises concentrating the solution of the compound of formula 4 to obtain the compound in solid form. Such concentration methods may be, for example, distillation of the solvent under reduced pressure (e.g., vacuum) by natural evaporation of the solvent. The solid form in which the compound of formula 4 is obtained may be, for example, crystalline or semi-crystalline, may be of a higher purity than before dissolution in an organic solvent, and then concentrated under acid conditions.

Thus, the present invention also relates to a method for purifying the compound of formula 4.

In some embodiments, the method comprises first dissolving the compound of formula 4 in an organic solvent to obtain a solution thereof, adding an acid to the solution of the compound of formula 4, and then concentrating the solution of the compound of formula 4 to obtain the compound in solid form. Examples of suitable organic solvents, acids, and solid forms have been provided above.

The invention further features compounds of formula 1a,

wherein R is₁Is P₂-；

P₂-is

R₅Is H, aliphatic, cycloaliphatic, heterocycloaliphatic, aryl or heteroaryl; each optionally substituted, in addition to H, with one or more substituents each independently selected from Group J consisting of: halo, cycloaliphatic, aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl, nitro, cyano, amido, amino, sulfonyl, sulfinyl, thio, sulfone, ureido, thioureido, sulfamoyl, sulfonamide, oxo, carboxy, carbamoyl, cycloaliphatic oxy, heterocycloaliphatic oxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, alkoxycarbonyl, alkylcarbonyloxy, and hydroxy;

R₈is H or a protecting group; and

R₃is an optionally substituted alkyl group.

In some embodiments, R₃Is a tert-butyl group.

In some other embodiments, P₂-is

In some further embodiments, P₂-is

The invention further relates to compounds of formula 1a as shown above, wherein

R₁Is P₃-L₂-P₂-；

P₃-L₂-P₂-is

R₆is optionally substituted aliphatic, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted phenyl; or R₅And R₆Together with the atoms to which they are attached, may form a 5-to 7-membered optionally substituted monocyclic heterocycle, or a 6-to 12-membered optionally substituted bicyclic heterocycle, wherein each heterocycle optionally contains a substituent selected from-O-, -S-, or-NR^X-a further heteroatom of (a);

R₇is H, optionally substituted aliphatic, optionally substituted heteroalkyl, optionally substituted heteroaryl, or optionally substituted phenyl; or

R₇And R₆Together with the atoms to which they are attached, may form a 5-to 7-membered optionally substituted monocyclic heterocycle, a 5-to 7-membered optionally substituted monocyclic aryl, a 6-to 12-membered optionally substituted bicyclic heterocycle or a 6-to 12-membered optionally substituted bicyclic aryl, wherein each heterocycle or aryl ring optionally contains a substituent selected from-O-, -S-or-NR^X-a further heteroatom of (a); or

R₈is H or a protecting group; and

R₃is an optionally substituted alkyl group.

In some embodiments, R₃Is a tert-butyl group.

In some embodiments, P₃-L₂-P₂-is

The compounds 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] nonane and 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] non-9-one are also within the scope of the present invention.

In one aspect, the invention includes a process for preparing a racemic mixture of cis-and trans-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acids of formula 7

The process comprises the step of carboxylation of an azabicyclooctane of formula 6,

wherein R' is C_1-5An alkyl group.

In one embodiment, the carboxylation step comprises forming the 2-anion of the compound of formula 6 in the presence of a complexing agent,

and treating the 2-anion with carbon dioxide to produce a racemic mixture of the formula 7 trans-/cis-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid.

In another embodiment, the 2-anion is prepared by treating a compound of formula 6 with a strong lithium base in the presence of a complexing agent and an aprotic solvent.

In a further embodiment, the base is sec-butyllithium.

In a further embodiment, the complexing agent is tetramethylethylenediamine, tetraethylethylenediamine, tetramethyl-1, 2-cyclohexyldiamine, sparteine, or 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] nonane.

In one embodiment, the complexing agent is tetramethylethylenediamine, tetraethylethylenediamine, tetramethyl-1, 2-cyclohexyldiamine, or 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] nonane.

In one embodiment, the trans-/cis-ratio is 1: 1.

In another embodiment, the trans-/cis-ratio is 60: 40.

In another embodiment, the trans-/cis-ratio is 80: 20.

In another embodiment, the trans-/cis-ratio is 90: 10.

In yet another embodiment, the trans-/cis-ratio is greater than 98: 2.

In one embodiment, the complexing agent is D-sparteine.

In one embodiment, the lithium base is sec-butyllithium and the complexing agent is 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] nonane to provide a mixture of racemic trans-/cis-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid of formula 7, wherein the trans-/cis-ratio is greater than 90: 10.

In a further embodiment, the trans-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid is trans-N-tert-butoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid.

In one aspect, the invention includes a process for preparing a compound of formula 4

4

The method comprises the following steps:

i) providing N-alkoxycarbonyl-3-azabicyclo [3.3.0] octane;

v) forming a salt with an optically active amine;

vi) crystallizing the salt;

vii) esterifying the acid provided in step vi);

ix) reacting the bicyclic amino ester of step viii) with a protected amino acid of formula 26,

x) removing the protecting group Z of step ix) the amide-ester to yield the amino compound of formula 28;

xii) removing the protecting group Z of the tripeptide of formula 30 to yield a free amino-tripeptide of formula 31;

In one embodiment, the oxidizing agent used in step xvi) is sodium hypochlorite and the oxidation is carried out in the presence of 2, 2, 6, 6-tetramethylpiperidinyloxy free radical (TEMPO).

In another embodiment, the oxidizing agent used in step xvi) is 1, 1-dihydro-1, 1, 1-triacetoxy-1, 2-benziodozol-3 (1H) -one.

In one embodiment, the method further comprises dissolving the compound of formula 4 in an organic solvent to obtain a solution of the compound of formula 4, and then adding an acid to the solution.

In a further embodiment, the organic solvent is dichloromethane and the acid is acetic acid.

In another embodiment, the method further comprises concentrating the solution of the compound of formula 4 to obtain the compound in solid form.

In one aspect, the invention includes a method of purifying a compound of formula 4, the method comprising:

4

i) dissolving the compound of formula 4 in an organic solvent to obtain a solution of the compound of formula 4,

ii) adding an acid to the solution of the compound of formula 4, and

iii) concentrating the solution of the compound of formula 4 to obtain the compound in solid form.

In one embodiment, the organic solvent is dichloromethane and the acid is acetic acid.

In one aspect, the invention includes a process for preparing a compound of formula 8

The process comprises reacting a 6 azabicyclooctane

Carboxylation to obtain a racemic mixture of cis-and trans-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acids of formula 7

Balancing the trans-/cis-mixture of the compounds of formula 7 in the presence of a suitable base to produce a trans-dominant-cis racemic acid of formula 8, wherein the trans-/cis-ratio in the compound of formula 8 is greater than 80: 20 and R' is C_1-5An alkyl group.

In one embodiment, the process further comprises balancing a trans-/cis-mixture of the compounds of formula 7 in the presence of a suitable base to produce a trans-predominately cis racemic acid of formula 8, wherein the trans-/cis-ratio is greater than 90: 10.

In another embodiment, the process further comprises balancing the trans-/cis-mixture of formula 7 in the presence of a suitable base to produce a trans-predominately cis racemic acid of formula 8, wherein the trans-/cis-ratio is greater than 98: 2.

In one embodiment, the base is lithium hexamethyldisilazide, lithium diisopropylamide, or lithium 2, 2, 6, 6-tetramethylpiperidine.

In further embodiments, wherein the base is lithium hexamethyldisilazide.

In one aspect, the invention includes a process for preparing (1S, 2S, 3R) trans-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid comprising carboxylating an azabicyclooctane of formula 6

To give a racemic mixture of cis-and trans-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acids of formula 7

Balancing the trans-/cis-mixture of the compound of formula 7 in the presence of a suitable base to produce a trans-dominant-cis racemic acid of formula 8

Wherein the trans-/cis-ratio in the compound of formula 8 is greater than 80: 20; and isolation of racemic trans-N-alkoxycarbonyl-octahydrocyclopenta [ c]Pyrrole-1-carboxylic acid mixtures wherein R' is C_1-5An alkyl group.

In one embodiment, the separation comprises the steps of:

i) forming a salt with an optically active base; and

ii) crystallizing the salt formed by step i) to provide the optically active salt of formula 9.

In a further embodiment, the optically active base is (R) α -aminoethylbenzene.

In a further embodiment, the optically active base is (S)1, 2, 3, 4-tetrahydro-1-naphthylamine.

In one aspect, the invention includes a process for preparing a compound of formula 1, the process comprising carboxylating an azabicyclooctane of formula 6

Wherein the trans-/cis-ratio in the compound of formula 8 is greater than 80: 20; forming a salt with an optically active base; crystallizing the salt formed in the previous step to provide the optically active salt of formula 9

The carboxylic acid of formula 9 is substituted with a compound containing R₃Esterification of a compound of the group; and removal of the-COOR' protecting group to yield the compound of formula 1

Wherein R' is C_1-5Alkyl radical, R₃Is an optionally substituted alkyl or aralkyl group.

In one embodiment, R₃Is a tert-butyl group.

The method comprises the following steps:

i) providing a compound of formula 7 prepared by the process described in claim 30;

ii) forming a salt with an optically active amine;

iii) crystallizing the salt;

iv) esterifying the acid provided in step iii);

v) removal of the N-alkoxycarbonyl group to yield (1S, 3aR, 6aS) -tert-butyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid tert-butyl ester;

vi) contacting the bicyclic amino ester of step v) with a protected amino acid of formula 26,

26

vii) removing the protecting group Z from the amide-ester of step vi) to produce an amino compound of formula 28;

viii) reacting an amino compound of formula 28 with a protected amino acid of formula 29

Reacting in the presence of a coupling reagent to produce a tripeptide of formula 30;

ix) removing the protecting group Z from the tripeptide of formula 30 to yield a free amino-tripeptide of formula 31;

x) reacting the amino-tripeptide of formula 31 with pyrazine-2-carboxylic acid in the presence of a coupling reagent to yield an amide-tripeptide ester of formula 33;

xi) hydrolyzing the ester of the amide-tripeptide ester of formula 33 to yield the amide-tripeptide acid of formula 34;

xii) reaction of an amide-tripeptide acid of formula 34 with an amino hydroxy-amide of formula 18

xiii) oxidizing the hydroxy group of formula 35 to yield the compound of formula 4.

The method comprises the following steps:

i) providing N-alkoxycarbonyl-3-azabicyclo [3.3.0] octane;

v) forming a salt with an optically active amine;

vi) crystallizing the salt;

vii) esterifying the acid provided in step vi);

xv) reacting an amide-tripeptide acid of formula 34 with an aminohydroxy-amide of formula 43

Reacting in the presence of a coupling reagent to produce a hydroxy-tetrapeptide of formula 36; and

xvi) oxidizing the hydroxy group of formula 36 to produce the compound of formula 4.

The method comprises the following steps:

i) providing N-alkoxycarbonyl-3-azabicyclo [3.3.0] octane;

v) forming a salt with an optically active amine;

vi) crystallizing the salt;

vii) esterifying the acid provided in step vi);

xv) reacting an amide-tripeptide acid of formula 34 with an aminohydroxy-amide of formula 44

Reacting in the presence of a coupling reagent to produce a hydroxy-tetrapeptide of formula 45; and

xvi) oxidizing the hydroxy group of formula 45 to produce the compound of formula 4.

In one embodiment, the method further comprises concentrating the solution of the compound of formula 4 to obtain the compound in solid form.

In one aspect, the invention includes compounds of formula 4 prepared by any of the methods described herein.

In one aspect, the invention includes compounds of formula 7 prepared by a process for preparing a racemic mixture of cis-and trans-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acids of formula 7 as described herein.

In one aspect, the invention includes a compound of formula 8 prepared by a process for preparing a compound of formula 8 as described herein.

In one aspect, the invention includes compounds of formula 9 prepared by the separation process of a racemic trans-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid mixture described herein.

In one aspect, the invention includes a compound of formula 1 prepared by a process for preparing a compound of formula 1 by carboxylation of an azabicyclooctane of formula 6as described herein.

Detailed Description

I. Definition of

For the purposes of the present invention, the chemical elements are specified according to the periodic table of the elements (CAS version, handbook of Chemistry and Physics, 75 th edition). In addition, the general principles of Organic Chemistry are described by Thomas Sorrell in Organic Chemistry, University scientific Books, Sausaltio (1999), and by M.B. Smith and J.March in advanced Organic Chemistry, 5 th edition, John Wiley & Sons, New York (2001), the entire contents of which are incorporated herein by reference.

As described herein, the compounds of the invention may be optionally substituted with one or more substituents, such as those generally set forth above, or exemplified by a particular class, subclass, or species of the invention.

The term "aliphatic" as used herein encompasses the terms alkyl, alkenyl and alkynyl, each of which is optionally substituted as set forth below.

As used herein, an "alkyl" group refers to a saturated aliphatic hydrocarbon group containing 1 to 8 (e.g., 1 to 6 or 1 to 4) carbon atoms. The alkyl group may be straight-chain or branched. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-heptyl, or 2-ethylhexyl. Alkyl groups may be substituted (i.e., optionally substituted) with one or more substituents selected from group J ("GroupJ"), consisting of: halo, cycloaliphatic (e.g., cycloalkyl or cycloalkenyl), heterocycloaliphatic (e.g., heterocycloalkyl or heterocycloalkenyl), aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl (e.g., (aliphatic) carbonyl, (cycloaliphatic) carbonyl, or (heterocycloaliphatic) carbonyl), nitro, cyano, amido (e.g., (cycloalkylalkyl) carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl) carbonylamino, (heterocycloalkylalkyl) carbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylaminoalkylaminocarbonyl, cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl, arylaminocarbonyl, or heteroarylaminocarbonyl), amino (e.g., aliphatic amino, cycloaliphatic amino, or heterocycloaliphatic amino), sulfonyl (e.g., aliphatic-SO-carbonyl)₂-), sulfinyl, thio, sulfone, ureido, thioureido, sulfamoyl,Sulfonamide, oxo, carboxy, carbamoyl, cycloaliphatic oxy, heterocycloaliphatic oxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, alkoxycarbonyl, alkylcarbonyloxy, and hydroxy. Some examples of substituted alkyl groups include, without limitation, carboxyalkyl (such as HOOC-alkyl, alkoxycarbonylalkyl, and alkylcarbonyloxyalkyl), cyanoalkyl, hydroxyalkyl, alkoxyalkyl, acylalkyl, aralkyl, (alkoxyaryl) alkyl, (sulfonylamino) alkyl (such as (alkyl-SO)₂-amino) alkyl), aminoalkyl, amidoalkyl, (cycloaliphatic) alkyl or haloalkyl.

As used herein, an "alkenyl" group refers to an aliphatic carbon group containing 2 to 8 (e.g., 2 to 6 or 2 to 4) carbon atoms and at least one double bond. Like alkyl groups, alkenyl groups may be straight-chain or branched. Examples of alkenyl groups include, but are not limited to, allyl, isoprenyl, 2-butenyl, and 2-hexenyl. The alkenyl Group may be optionally substituted with one or more substituents selected from Group J, such as halo, cycloaliphatic (e.g., cycloalkyl or cycloalkenyl), heterocycloaliphatic (e.g., heterocycloalkyl or heterocycloalkenyl), aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl (e.g., (aliphatic) carbonyl, (cycloaliphatic) carbonyl, or (heterocycloaliphatic) carbonyl), nitro, cyano, amido (e.g., (cycloalkylalkyl) carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl) carbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylaminoalkylaminocarbonyl, cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl, arylaminocarbonyl or heteroarylaminocarbonyl), amino (e.g., aliphatic amino, cycloaliphatic amino, heterocycloaliphatic amino, or aliphatic sulfonylamino), Sulfonyl (e.g. alkyl-SO)₂-, cycloaliphatic-SO₂-or aryl-SO₂-), sulfinyl, thio, sulfone, ureido, thioureido, sulfamoyl, sulfonamide, oxo, carboxy, carbamoyl, cycloaliphatic oxy, heterocycloaliphatic oxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, alkoxycarbonyl, alkylcarbonyloxy or hydroxy. Without limitationSome examples of substituted alkenyl include cyanoalkenyl, alkoxyalkenyl, acylalkenyl, hydroxyalkenyl, aralkenyl, (alkoxyaryl) alkenyl, (sulfonylamino) alkenyl (such as (alkyl-SO)₂-amino) alkenyl), aminoalkenyl, amidoalkenyl, (cycloaliphatic) alkenyl, or haloalkenyl.

As used herein, an "alkynyl" group refers to an aliphatic carbon group containing 2 to 8 (e.g., 2 to 6 or 2 to 4) carbon atoms and at least one triple bond. The alkynyl group may be linear or branched. Examples of alkynyl groups include, but are not limited to, propargyl and butynyl. Alkynyl may be optionally substituted with one or more substituents selected from GroupJ, such as aroyl, heteroaroyl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, nitro, carboxy, cyano, halo, hydroxy, sulfo, mercapto, thio (such as aliphatic or cycloaliphatic thio), sulfinyl (such as aliphatic sulfinyl or cycloaliphatic sulfinyl), sulfonyl (such as aliphatic-SO)₂-, aliphatic amino-SO₂Or cycloaliphatic-SO₂-), acylamino (e.g.aminocarbonyl, alkylaminocarbonyl, alkylcarbonylamino, cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl, cycloalkylcarbonylamino, arylaminocarbonyl, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl) carbonylamino, (cycloalkylalkyl) carbonylamino, heteroaralkylcarbonylamino, heteroarylcarbonylamino or heteroarylaminocarbonyl), ureido, thioureido, sulfamoyl, sulfonamido, alkoxycarbonyl, alkylcarbonyloxy, cycloaliphatic, heterocycloaliphatic, aryl, heteroaryl, acyl (e.g. (cycloaliphatic) carbonyl or (heterocycloaliphatic) carbonyl), amino (e.g. aliphatic amino), sulfone, oxo, carboxyl, carbamoyl, (cycloaliphatic) oxy, (heterocycloaliphatic) oxy or (heteroaryl) alkoxy.

As used herein, "amido" encompasses both "aminocarbonyl" and "carbonylamino". These terms, when used alone or in combination with another group, when used terminally refer to an amido group such as-N (R)^X)-C(O)-R^Yor-C (O) -N (R)^X)₂(ii) a When used inWhen substituted, they refer to amide groups such as-C (O) -N (R)^X) -or-N (R)^X) -C (O) -, wherein R^XAnd R^YAs defined below. Examples of acylamino groups include alkylamido (such as alkylcarbonylamino or alkylaminocarbonyl), (heterocycloaliphatic) acylamino, (heteroaralkyl) acylamino, (heteroaryl) acylamino, (heterocycloalkyl) alkylamido, arylamido, aralkylamido, (cycloalkyl) alkylamido or cycloalkylamido.

As used herein, an "amino" group refers to-NR^XR^YWherein R is^XAnd R^YEach independently is hydrogen, aliphatic, cycloaliphatic, (cycloaliphatic) aliphatic, aryl, araliphatic, heterocycloaliphatic, (heterocycloaliphatic) aliphatic, heteroaryl, carboxy, thio, sulfinyl, sulfonyl, (aliphatic) carbonyl, (cycloaliphatic) carbonyl, ((cycloaliphatic) aliphatic) carbonyl, arylcarbonyl, (araliphatic) carbonyl, (heterocycloaliphatic) carbonyl, ((heterocycloaliphatic) aliphatic) carbonyl, (heteroaryl) carbonyl, or (heteroarylaliphatic) carbonyl, each of which is as defined herein and is optionally substituted. Examples of the amino group include an alkylamino group, a dialkylamino group, or an arylamino group. When the term "amino" is not a terminal group (e.g., alkylcarbonylamino), with-NR^X-represents. R^XHave the same meaning as defined above.

As used herein, an "aryl" group, used alone or as part of a larger moiety such as "aralkyl", "aralkoxy", or "aryloxyalkyl", refers to a monocyclic ring (e.g., phenyl); bicyclic (e.g., indenyl, naphthyl, tetrahydronaphthyl, tetrahydroindenyl); and tricyclic (e.g., fluorenyl, tetrahydrofluorenyl, or tetrahydroanthracenyl, anthracenyl) ring systems, wherein the monocyclic ring system is aromatic or at least one ring is aromatic in a bicyclic or tricyclic ring system. Bicyclic and tricyclic groups include benzo-fused 2-to 3-membered carbocyclic rings. For example, the benzo-fused group includes two or more C' s_4-8Phenyl fused to a carbocyclic moiety. Aryl is optionally substituted with one or more substituents, such as aliphatic (e.g., alkyl, alkenyl, or alkynyl); cycloaliphatic; (cycloaliphatic) aliphatic; (ii) heterocycloaliphatic; (heterocycloaliphatic) aliphatic; an aryl group; a heteroaryl group; an alkoxy group; (cycloaliphatic)) An oxy group; (heterocycloaliphatic) oxy; an aryloxy group; a heteroaryloxy group; (araliphatic) oxy; (heteroarylaliphatic) oxy; aroyl; a heteroaroyl group; an amino group; oxo (on a non-aromatic carbocyclic ring of a benzo-fused bicyclic or tricyclic aryl); a nitro group; a carboxyl group; an amido group; acyl (e.g., aliphatic; (cycloaliphatic) carbonyl; (cycloaliphatic) aliphatic; (araliphatic) carbonyl; (heterocycloaliphatic) aliphatic; or (heteroarylaliphatic) carbonyl); sulfonyl (e.g. aliphatic-SO)₂-or amino-SO₂-) according to the formula (I); sulfinyl (e.g., aliphatic-S (O) -or cycloaliphatic-S (O)); thio (e.g., aliphatic-S-); a cyano group; a halo group; a hydroxyl group; a mercapto group; a sulfone group; a urea group; a thiourea group; a sulfamoyl group; a sulfonamide group; or a carbamoyl group. Alternatively, the aryl group may be unsubstituted.

Non-limiting examples of substituted aryl groups include haloaryl groups (e.g., mono-, di- (e.g., p, m-dihaloaryl) or (trihalo) aryl); (carboxy) aryl (such as (alkoxycarbonyl) aryl, ((aralkyl) carbonyloxy) aryl, or (alkoxycarbonyl) aryl); (amido) aryl (such as (aminocarbonyl) aryl, ((alkylamino) alkyl) aminocarbonyl) aryl, (alkylcarbonyl) aminoaryl, (arylaminocarbonyl) aryl or (((heteroaryl) amino) carbonyl) aryl); aminoaryl groups (such as ((alkylsulfonyl) amino) aryl or ((dialkyl) amino) aryl); (cyanoalkyl) aryl; (alkoxy) aryl; (sulfamoyl) aryl (e.g., (sulfamoyl) aryl); (alkylsulfonyl) aryl; (cyano) aryl; (hydroxyalkyl) aryl; ((alkoxy) alkyl) aryl; (hydroxy) aryl, ((carboxy) alkyl) aryl; ((dialkyl) amino) alkyl) aryl; (nitroalkyl) aryl; ((alkylsulfonyl) amino) alkyl) aryl; ((heterocycloaliphatic) carbonyl) aryl; ((alkylsulfonyl) alkyl) aryl; (cyanoalkyl) aryl; (hydroxyalkyl) aryl; (alkylcarbonyl) aryl; an alkylaryl group; (trihaloalkyl) aryl; p-amino-m-alkoxycarbonylaryl; p-amino-m-cyanoaryl; p-halo-m-aminoaryl; or (meta- (heterocycloaliphatic) -ortho- (alkyl)) aryl.

As used herein, an "araliphatic" group, such as "aralkyl", refers toAliphatic radicals substituted by aryl radicals (e.g. C)_1-4Alkyl groups). "aliphatic", "alkyl" and "aryl" are defined herein. An example of an araliphatic such as aralkyl is benzyl.

As used herein, an "aralkyl" group refers to an alkyl group substituted with an aryl group (e.g., C)_1-4Alkyl groups). Both "alkyl" and "aryl" are defined above. An example of an aralkyl group is benzyl. An aralkyl group is optionally substituted with one or more substituents such as aliphatic (e.g., substituted or unsubstituted alkyl, alkenyl, or alkynyl groups including carboxyalkyl, hydroxyalkyl, or haloalkyl such as trifluoromethyl), cycloaliphatic (e.g., substituted or unsubstituted cycloalkyl or cycloalkenyl), (cycloalkyl) alkyl, heterocycloalkyl, (heterocycloalkyl) alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, amido (e.g., aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkylalkyl) carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl) carbonylamino, (heterocycloalkylalkyl) carbonylamino, and the like, Heteroarylcarbonylamino or heteroarylalkylcarbonylamino), cyano, halo, hydroxy, acyl, mercapto, alkylthio, sulfone, ureido, thioureido, sulfamoyl, sulfonamido, oxo, or carbamoyl.

As used herein, a "bicyclic ring system" includes 8-to 12- (e.g., 9, 10, or 11) membered structures forming 2 rings, wherein the 2 rings have at least one common atom (e.g., 2 common atoms). Bicyclic ring systems include bicyclic aliphatic (e.g., bicycloalkyl or bicycloalkenyl), bicyclic heteroaliphatic, bicyclic aryl, and bicyclic heteroaryl.

As used herein, "cycloaliphatic" groups encompass "cycloalkyl" and "cycloalkenyl," each of which is optionally substituted as set forth below.

As used herein, a "cycloalkyl" group refers to a saturated carbocyclic mono-or bicyclic (fused or otherwise) ring of 3 to 10 (e.g., 5 to 10) carbon atomsBridge) the loops. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, norbornyl, cubic alkyl, octahydro-indenyl, decahydro-naphthyl, bicyclo [3.2.1]Octyl, bicyclo [2.2.2]Octyl, bicyclo [3.3.1]Nonyl, bicyclo [3.3.2.]Decyl, bicyclo [2.2.2]Octyl, adamantyl, azacycloalkyl or ((aminocarbonyl) cycloalkyl. "cycloalkenyl" groups, as used herein, refer to non-aromatic carbocyclic rings of 3 to 10 (e.g., 4 to 8) carbon atoms having one or more double bonds. Examples of cycloalkenyl include cyclopentenyl, 1, 4-cyclohex-di-alkenyl, cycloheptenyl, cyclooctenyl, hexahydro-indenyl, octahydro-naphthyl, cyclohexenyl, cyclopentenyl, bicyclo [2.2.2]Octenyl or bicyclo [3.3.1]Nonenyl. Cycloalkyl or cycloalkenyl groups may be optionally substituted by one or more substituents selected from Group J, such as aliphatic (e.g. alkyl, alkenyl or alkynyl), cycloaliphatic, (cycloaliphatic) aliphatic, heterocycloaliphatic, (heterocycloaliphatic) aliphatic, aryl, heteroaryl, alkoxy, (cycloaliphatic) oxy, (heterocycloaliphatic) oxy, aryloxy, heteroaryloxy, (araliphatic) oxy, (heteroarylaliphatic) oxy, aroyl, heteroaroyl, amino, acylamino (e.g. (aliphatic) carbonylamino, (cycloaliphatic) carbonylamino, (aryl) carbonylamino, (araliphatic) carbonylamino, (heterocycloaliphatic) carbonylamino, (heteroaryl) carbonylamino or ((heteroarylaliphatic) carbonylamino), nitro, carboxy (e.g. HOOC-), Alkoxycarbonyl or alkylcarbonyloxy), acyl ((e.g. (cycloaliphatic) carbonyl, (cycloaliphatic) aliphatic) carbonyl, (araliphatic) carbonyl, (heterocycloaliphatic) carbonyl, ((heterocycloaliphatic) aliphatic) carbonyl or (heteroarylaliphatic) carbonyl), cyano, halo, hydroxy, mercapto, sulfonyl (e.g. alkyl-SO)₂And aryl-SO₂-), sulfinyl ((e.g., alkyl-S (O)) -, thio (e.g., alkyl-S-), sulfone, ureido, thioureido, sulfamoyl, sulfonamide, oxo, or carbamoyl.

As used herein, "cyclic moiety" includes cycloaliphatic, heterocycloaliphatic, aryl, or heteroaryl, each of which has been defined above.

As used herein, the term "heterocycloaliphatic" encompasses heterocycloalkyl and heterocycloalkenyl, each of which is optionally substituted as set forth below.

As used herein, a "heterocycloalkyl" group refers to a 3-10 membered mono-or bicyclic (fused or bridged) (e.g., 5-to 10-membered mono-or bicyclic) saturated ring structure in which one or more ring atoms is a heteroatom (e.g., N, O, S or a combination thereof). Examples of heterocycloalkyl include piperidinyl, piperazinyl, tetrahydropyranyl, tetrahydrofuranyl, 1, 4-dioxolanyl, 1, 4-dithianyl, 1, 3-dioxolanyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiomorpholinyl, octahydrobenzofuranyl, octahydrochromenyl, octahydrothiochromenyl, octahydroindolyl, octahydropyridinyl, decahydroquinolinyl, octahydrobenzo [ b ] thienyl, 2-oxa-bicyclo [2.2.2] octyl, 1-aza-bicyclo [2.2.2] octyl, 3-aza-bicyclo [3.2.1] octyl, and 2, 6-dioxa-tricyclo [3.3.1.03, 7] nonyl. Monocyclic heterocycloalkyl groups can be fused to a phenyl moiety such as tetrahydroisoquinoline. A "heterocycloalkenyl" group, as used herein, refers to a mono-or bicyclic (e.g., 5-to 10-membered mono-or bicyclic) non-aromatic ring structure having one or more double bonds in which one or more ring atoms is a heteroatom (e.g., N, O or S). Monocyclic and bicyclic heteroaliphats are numbered according to standard chemical nomenclature.

The heterocycloalkyl or heterocycloalkenyl radical may be optionally substituted by one or more substituents selected from the Group J, such as aliphatic (e.g. alkyl, alkenyl or alkynyl), cycloaliphatic, (cycloaliphatic) aliphatic, heterocycloaliphatic, (heterocycloaliphatic) aliphatic, aryl, heteroaryl, alkoxy, (cycloaliphatic) oxy, (heterocycloaliphatic) oxy, aryloxy, heteroaryloxy, (araliphatic) oxy, (heteroarylaliphatic) oxy, aroyl, heteroaroyl, amino, acylamino (e.g. (aliphatic) carbonylamino, (cycloaliphatic) carbonylamino, (aryl) carbonylamino, (araliphatic) carbonylamino, (heterocycloaliphatic) carbonylamino, ((heterocycloaliphatic) carbonylamino, (heteroaryl) carbonylamino or (heteroarylaliphatic) carbonylamino), nitro, carboxy (e.g. HOOC-), Alkoxycarbonyl or alkylcarbonyloxy), acyl ((e.g., (cycloaliphatic) carbonyl, ((cycloaliphatic) aliphatic) carbonyl, (araliphatic) carbonyl, (heterocycloaliphatic) carbonyl, ((heterocycloaliphatic) aliphatic) carbonyl, or (heteroarylaliphatic) carbonyl), nitro, cyano, halo, hydroxy, mercapto, sulfonyl (e.g., alkylsulfonyl or arylsulfonyl), sulfinyl (e.g., alkylsulfinyl), thio (e.g., alkylthio), sulfone, ureido, thioureido, sulfamoyl, sulfonamide, oxo, or carbamoyl)).

A "heteroaryl" group, as used herein, refers to a monocyclic, bicyclic, or tricyclic ring system having 4-15 ring atoms, wherein one or more ring atoms is a heteroatom (e.g., N, O, S or a combination thereof), wherein the monocyclic ring system is aromatic, or at least one ring is aromatic in the bicyclic or tricyclic ring system. Heteroaryl groups include benzo-fused ring systems having 2-3 rings. For example, benzo-fused groups include benzo groups fused to 1 or 2 4-8 membered heterocycloaliphatic moieties (e.g., indolizinyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo [ b ] furanyl, benzo [ b ] thienyl, quinolinyl, or isoquinolinyl). Examples of some heteroaryl groups are azetidinyl, pyridyl, 1H-indazolyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, tetrazolyl, benzofuryl, isoquinolyl, benzothiazolyl, xanthene, thioxanthene, phenothiazine, indoline, benzo [1, 3] dioxole, benzo [ b ] furyl, benzo [ b ] thienyl, indazolyl, benzimidazolyl, benzothiazolyl, pyrrolyl (puryl), cinnolinyl, quinolinyl, quinazolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, isoquinolinyl, 4H-quinolizinyl, benzo-1, 2, 5-thiadiazolyl or 1, 8-naphthyridinyl.

Monocyclic heteroaryl includes, without limitation, furyl, thienyl, 2H-pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1, 3, 4-thiadiazolyl, 2H-pyranyl, 4-H-pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazolyl, pyrazinyl or 1, 3, 5-triazinyl. Monocyclic heteroaryls are numbered according to standard chemical nomenclature.

Bicyclic heteroaryls include, without limitation, indolizinyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo [ b ] furanyl, benzo [ b ] thienyl, quinolinyl, isoquinolinyl, indolizinyl, isoindolyl, indolyl, benzo [ b ] furanyl, benzo [ b ] thienyl, indazolyl, benzimidazolyl, benzothiazolyl, purinyl, 4H-quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1, 8-naphthyridinyl, or pteridinyl. Bicyclic heteroaryls are numbered according to standard chemical nomenclature.

Heteroaryl is optionally substituted with one or more substituents, such as aliphatic (e.g., alkyl, alkenyl, or alkynyl); cycloaliphatic; (cycloaliphatic) aliphatic; (ii) heterocycloaliphatic; (heterocycloaliphatic) aliphatic; an aryl group; a heteroaryl group; an alkoxy group; (cycloaliphatic) oxy; (heterocycloaliphatic) oxy; an aryloxy group; a heteroaryloxy group; (araliphatic) oxy; (heteroarylaliphatic) oxy; aroyl; a heteroaroyl group; an amino group; oxo (on a non-aromatic carbocyclic or heterocyclic ring of a bicyclic or tricyclic heteroaryl); a carboxyl group; an amido group; acyl (e.g., aliphatic; (cycloaliphatic) carbonyl; (cycloaliphatic) aliphatic; (araliphatic) carbonyl; (heterocycloaliphatic) aliphatic; or (heteroarylaliphatic) carbonyl); sulfonyl (such as aliphatic sulfonyl or aminosulfonyl); sulfinyl (e.g., aliphatic sulfinyl); sulfur radicals (such as aliphatic sulfur radicals); a nitro group; a cyano group; a halo group; a hydroxyl group; a mercapto group; a sulfone group; a urea group; a thiourea group; a sulfamoyl group; a sulfonamide group; or a carbamoyl group. Alternatively, the heteroaryl group may be unsubstituted.

Non-limiting examples of substituted heteroaryl groups include (halo) heteroaryl groups (e.g., mono-and di- (halo) heteroaryl); (carboxy) heteroaryl (e.g., (alkoxycarbonyl) heteroaryl); a cyanoheteroaryl group; aminoheteroaryl groups (such as ((alkylsulfonyl) amino) heteroaryl and ((dialkyl) amino) heteroaryl); (amido) heteroaryl (e.g., aminocarbonylheteroaryl, ((alkylcarbonyl) amino) heteroaryl, ((((alkyl) amino) alkyl) aminocarbonyl) heteroaryl, (((heteroaryl) amino) carbonyl) heteroaryl, ((heterocycloaliphatic) carbonyl) heteroaryl, or ((alkylcarbonyl) amino) heteroaryl); (cyanoalkyl) heteroaryl; (alkoxy) heteroaryl; (sulfamoyl) heteroaryl (e.g., (aminosulfonyl) heteroaryl); (sulfonyl) heteroaryl ((e.g., (alkylsulfonyl) heteroaryl); (hydroxyalkyl) heteroaryl; (alkoxyalkyl) heteroaryl; (hydroxy) heteroaryl; (carboxy) alkyl) heteroaryl; ((dialkyl) amino) alkyl) heteroaryl; (heterocycloaliphatic) heteroaryl; (cycloaliphatic) heteroaryl; (nitroalkyl) heteroaryl; ((alkylsulfonyl) amino) alkyl) heteroaryl; (alkylsulfonyl) alkyl) heteroaryl; (cyanoalkyl) heteroaryl; (acyl) heteroaryl (e.g., (alkylcarbonyl) heteroaryl; (alkyl) heteroaryl; (haloalkyl) heteroaryl; (trihaloalkylheteroaryl) (e.g., (trihaloalkylheteroaryl).

"Heteroaraliphatic" (e.g., heteroaralkyl), as used herein, refers to an aliphatic group (e.g., C) substituted with a heteroaryl group_1-4Alkyl groups). "aliphatic", "alkyl" and "heteroaryl" have been defined above.

A "heteroaralkyl" group, as used herein, refers to an alkyl group (e.g., C) substituted with a heteroaryl group_1-4Alkyl groups). Both "alkyl" and "heteroaryl" have been defined above. Heteroarylalkyl is optionally substituted with one or more substituents, such as alkyl (including carboxyalkyl, hydroxyalkyl and haloalkyl such as trifluoromethyl), alkenyl, alkynyl, cycloalkyl, (cycloalkyl) alkyl, heterocycloalkyl, (heterocycloalkyl) alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkylalkyl) carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl) carbonylamino, heteroarylcarbonylamino, heteroaralylcarbonylamino, cyano, halo, hydroxy, acyl, mercapto, halo, cyano, hydroxy, acyl, mercapto, cyano, hydroxy, mercapto, hydroxy, mercapto, hydroxy, Alkylthio, sulfone, ureido, thioureido, sulfamoyl, sulfonamide, oxo, or carbamoyl.

As used herein, an "acyl" group refers to acyl or R^X-C (O) - (e.g. alkanes)radical-C (O) -, also known as "alkylcarbonyl"), wherein R^XAnd "alkyl" have been defined above. Acetyl and pivaloyl are examples of acyl groups.

As used herein, "aroyl" or "heteroaroyl" refers to aryl-C (O) -or heteroaryl-C (O) -. The aryl and heteroaryl portions of the aroyl or heteroaroyl groups are optionally substituted as defined hereinbefore.

As used herein, "alkoxy" refers to an alkyl-O-group, wherein "alkyl" has been previously defined.

As used herein, "carbamoyl" refers to a compound having the structure-O-CO-NR^XR^Yor-NR^X-CO-O-R^ZWherein R is^XAnd R^YAs already defined above, R^ZMay be aliphatic, aryl, araliphatic, heterocycloaliphatic, heteroaryl or heteroarylaliphatic.

As used herein, "carboxy" when used as a terminal group refers to-COOH, -COOR^X、-OC(O)H、-OC(O)R^X(ii) a or-OC (O) -or-C (O) O-when used as an internal group.

As used herein, a "haloaliphatic" group refers to an aliphatic group substituted with 1-3 halogens. For example, the term haloalkyl includes the group-CF₃。

As used herein, a "mercapto" group refers to-SH.

As used herein, a "sulfo" group, when used at the terminus, refers to-SO₃H or-SO₃R^Xor-S (O) when used internally₃-。

As used herein, a "sulfonamide" group, when used at the terminus, refers to the structure-NR^X-S(O)₂-NR^YR^ZWhen used internally, is referred to as-NR^X-S(O)₂-NR^Y-, wherein R^X、R^YAnd R^ZAs already defined above.

As used herein, a "sulfonamide" group, when used at a terminus, refers to a knotstructure-S (O)₂-NR^XR^Yor-NR^X-S(O)₂-R^Z(ii) a or-S (O) when used internally₂-NR^X-or-NR^X-S(O)₂-, wherein R^X、R^YAnd R^ZAs defined hereinabove.

As used herein, "thio" when used terminally means-S-R^XWhen used internally, refers to-S-, wherein R^XAs already defined above. Examples of sulfur radicals include aliphatic-S-, cycloaliphatic-S-, aryl-S-, and the like.

As used herein, "sulfinyl" when used terminally means-S (O) -R^XWhen used internally refers to-S (O) -, wherein R^XAs already defined above. Exemplary sulfinyl groups include aliphatic-s (o) -, aryl-s (o) -, (cycloaliphatic (aliphatic)) -s (o) -, cycloalkyl-s (o) -, heterocycloaliphatic-s (o) -, heteroaryl-s (o) -, and the like.

As used herein, a "sulfonyl" group, when used terminally, refers to-S (O)₂-R^XWhen used internally, means-S (O)₂-, wherein R^XAs already defined above. Exemplary sulfonyl groups include aliphatic-S (O)₂-, aryl-S (O)₂-, ((cycloaliphatic (aliphatic)) -S (O))₂-, cycloaliphatic-S (O)₂-, heterocycloaliphatic-S (O)₂-, heteroaryl-S (O)₂-, (cycloaliphatic (amido (aliphatic))) -S (O)₂-and the like.

As used herein, "sulfone group" when used at the terminus refers to-O-SO-R^Xor-SO-O-R^XWhen used internally means-O-S (O) -or-S (O) -O-, wherein R^XAs already defined above.

As used herein, a "halogen" or "halo" group refers to fluorine, chlorine, bromine, or iodine.

As used herein, "alkoxycarbonyl," which is encompassed by "carboxy," used alone or in combination with another group, refers to a group such as alkyl-O-c (O) -.

As used herein, an "alkoxyalkyl" group refers to an alkyl group such as alkyl-O-alkyl-, where alkyl has been defined above.

As used herein, a "carbonyl" group refers to-C (O) -.

As used herein, an "oxo" group refers to ═ O.

As used herein, an "aminoalkyl" group refers to the structure (R)^X)₂N-alkyl-.

As used herein, a "cyanoalkyl" group refers to the structure (NC) -alkyl-.

As used herein, a "ureido" group refers to the structure-NR^X-CO-NR^YR^ZThe "thioureido" group, when used at the terminus, refers to the structure-NR^X-CS-NR^YR^ZWhen used internally, is referred to as-NR^X-CO-NR^Y-or-NR^X-CS-NR^Y-, wherein R^X、R^YAnd R^ZAs already defined above.

As used herein, a "guanidine" group refers to the structure-N ═ C (N (R)^XR^Y))N(R^XR^Y) or-NR^X-C(＝NR^X)NR^XR^YWherein R is^XAnd R^YAs already defined above.

As used herein, an "amidino" group refers to the structure-C ═ (NR)^X)N(R^XR^Y) Wherein R is^XAnd R^YAs already defined above.

Generally, the term "ortho" refers to the position of a substituent on a group comprising two or more carbon atoms, wherein the substituent is attached to an adjacent carbon atom.

Generally, the term "geminal" refers to the position of a substituent on a group comprising two or more carbon atoms, wherein the substituent is attached to the same carbon atom.

The terms "terminal" and "internal" refer to the position of a group within a substituent. When the group appears no further to bond chemicallyThe group is terminal when it forms the end of a substituent to which the remainder is bonded. Carboxyalkyl, i.e. R^XO (O) C-alkyl is an example of a carboxyl group used at the end. A group is internal when it appears in the middle of a substituent, the end of which is bonded to the rest of the chemical structure. Alkylcarboxy (such as alkyl-C (O) -O-or alkyl-O-C (O) -) and alkylcarboxylaryl (such as alkyl-C (O) -O-aryl-or alkyl-O-C (O) -aryl-) are examples of carboxyl groups for the interior.

As used herein, "cyclic" groups include mono-, bi-and tri-cyclic ring systems, such as cycloaliphatic, heterocycloaliphatic, aryl or heteroaryl, each of which has been defined above.

As used herein, "bridged bicyclic ring system" refers to a bicyclic heterocyclic aliphatic ring system or bicyclic cycloaliphatic ring system in which the rings are bridged. Examples of bridged bicyclic ring systems include, but are not limited to, adamantyl, norbornyl, bicyclo [3.2.1]Octyl, bicyclo [2.2.2]Octyl, bicyclo [3.3.1]Nonyl, bicyclo [3.2.3]Nonyl, 2-oxabicyclo [2.2.2]Octyl, 1-azabicyclo [2.2.2]Octyl, 3-azabicyclo [3.2.1]Octyl and 2, 6-dioxa-tricyclo [3.3.1.0^3，7]Nonyl radical. The bridged bicyclic ring system may be optionally substituted with one or more substituents such as alkyl (including carboxyalkyl, hydroxyalkyl and haloalkyl such as trifluoromethyl), alkenyl, alkynyl, cycloalkyl, (cycloalkyl) alkyl, heterocycloalkyl, (heterocycloalkyl) alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkylalkyl) carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl) carbonylamino, (heterocycloalkylalkyl) carbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylamino, cyano, halo, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof, Hydroxyl, acyl, mercapto, alkylthio, sulfone, ureido, thioureido, sulfamoyl, sulfonamide, oxo, or carbamoyl.

As used herein, "aliphatic chain" refers to a branched or straight chain aliphatic group (e.g., alkyl, alkenyl, or alkynyl). The linear aliphatic chain has the structure- (CH)₂)_v-, where v is 1 to 6. A branched aliphatic chain is a straight aliphatic chain substituted with one or more aliphatic groups. The branched aliphatic chain has the structure- (CHQ)_v-, wherein Q is hydrogen or an aliphatic group; however, in at least one instance Q should be an aliphatic group. The term aliphatic chain includes alkyl, alkenyl and alkynyl chains, wherein alkyl, alkenyl and alkynyl are defined above.

The phrase "optionally substituted" is used interchangeably with the phrase "substituted or unsubstituted". As described herein, the compounds of the present invention may be optionally substituted with one or more substituents, such as those generally indicated above, or exemplified by particular classes, subclasses, and species of the invention. The variable R is as described herein₁、R₂And R₃And other variables, covering specific groups such as alkyl and aryl. Unless otherwise stated, the variable R₁、R₂And R₃Each of the specific groups of (a), and the other variables contained therein, may be optionally substituted with one or more substituents described herein. Each substituent of a particular group is further optionally substituted with 1-3 of: halo, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl, cycloaliphatic, heterocycloaliphatic, heteroaryl, haloalkyl, and alkyl. For example, the alkyl group may be substituted with an alkylthio group, which may be optionally substituted with 1 to 3 of: halo, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl, haloalkyl, and alkyl. As another example, the cycloalkyl moiety of the (cycloalkyl) carbonylamino group can be optionally substituted with 1-3 of the following: halo, cyano, alkoxy, hydroxy, nitro, haloalkyl, and alkyl. When 2 alkoxy groups are bound to the same atom or adjacent atoms, 2 alkoxy groups may form a ring together with the atom to which they are bound.

Generally, the term "substituted", whether or not after the term "optional", refers to a radical replacement of a hydrogen radical in a given structure with a radical designated a substituent. Specific substituents are described in the above definitions and in the following description of the compounds and examples thereof. Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents at each position may be the same or different. A ring substituent, such as heterocycloalkyl, may be joined to another ring, such as cycloalkyl, to form a spiro-bicyclic ring system, e.g., the two rings share a common atom. Combinations of substituents contemplated by the present invention are those that result in the formation of stable or chemically feasible compounds.

The phrase "stable or chemically feasible," as used herein, refers to a compound that is not substantially altered when subjected to conditions that allow its preparation, detection, and preferably its recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, a stable compound or chemically feasible compound is a compound that does not substantially change when stored for at least one week at a temperature of 40 ℃ or less in the absence of moisture or other chemically reactive conditions.

As used herein, an effective amount is defined as the amount required to deliver a therapeutic effect to a treated patient, typically depending on the age, body surface area, weight, and condition of the patient. Animal-to-human dose correlations (based on milligrams per square meter of body surface) are described in Freireich et al, cancer chemither. 219(1966). The body surface area can be roughly determined by the height and weight of the patient. See, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley, New York, 537 (1970). As used herein, "patient" refers to a mammal, including a human.

Unless otherwise stated, the structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational) forms of the structure; for example, the R and S configurations of each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Thus, the individual stereochemical isomers as well as enantiomers, diastereomers and geometries of the compounds of the present inventionMixtures of isomers (or conformers) are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention. In addition, unless otherwise stated, the structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, except for replacement of hydrogen by deuterium or tritium, or replacement of carbon by deuterium or tritium¹³C-or¹⁴Compounds having this structure other than C-rich carbon substitution are within the scope of the invention. Such compounds may be used, for example, as analytical tools or probes in bioassays.

As used herein, EDC is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide, HOBt is 1-hydroxybenzotriazole, HOSuc is N-hydroxysuccinimide, THF is tetrahydrofuran, TFA is trifluoroacetic acid, DCM is dichloromethane, DMAP is 4-dimethylaminopyridine, DIPEA is diisopropylethylamine, DMF is dimethylformamide, TFA is trifluoroacetic acid, CBZ is benzyloxycarbonyl, TEMPO is 2, 2, 6, 6-tetramethylpiperidinyloxy.

As used herein, the term "a" or "an" refers to,¹h NMR stands for proton nuclear magnetic resonance and TLC for thin layer chromatography.

Process and intermediates

In one embodiment, the present invention provides processes and intermediates for preparing compounds of formula 1, as outlined in scheme I.

Scheme I

Referring to scheme I, 3-azabicyclo [3.3.0] octane of formula 5 (R.Griot, Helv.Chim.acta., 42, 67, (1959) is converted to a suitable alkyl carbamate of formula 6, wherein R' is, e.g., t-butyl or isobutyl, using known methods, see, e.g., T.W.Greene and P.G.M.Wuts, Protective Groups in Organic Synthesis, 3 rd edition, John Wiley and Sons, Inc. (1999).

Carboxylation of N-alkoxycarbonyl-3-azabicyclo [3.3.0] octane of the formula 6 is achieved by first forming the 2-anion of the formula 6 in the presence of a chelating agent (for the formation of analogous anions see, for example, Daniel. J. Pippel et al, J. Org. chem., 1998, 63, 2; Donald J. Gallagher et al, J. Org. chem., 1995, 60(22), 7092. conk 7093; Shawn T. Kerrick et al, J.am. chem. Soc., 1991, 113(25), 9708. cona 9710; Donald J. Gallagher et al, J. Org. chem., 1995, 60(25), 8148. cona 8154; and Peter Be et al, J.am. soc., chem. Soc., 1994, 116(8), 3231. 3239. carbamic acid alkyl ester of the formula 6. carbamic acid anion of the formula 6 is shown by the scheme for example using a strong lithium-ethylene diamine, tetraethyl diamine (3-tetramethyl diamine, 7-t. ethylene diamine, or tetraethyl diamine (shown in the formula 1, 7), 7-diazabicyclo [3.3.1] nonane) in a suitable aprotic solvent. Suitable aprotic solvents include, for example, t-butyl methyl ether, tetrahydrofuran and dimethoxyethane. The 2-anion of formula 6 can then be treated with carbon dioxide to give a racemic mixture of trans-/cis-2-carboxylic acid of formula 7, wherein the trans-/cis-ratio is 30: 70, 40: 60, 50: 50, 60: 40, 80: 20, 90: 10, 95: 5 or greater than 98: 2.

In some embodiments, the complexing agent may be optically active, such as, for example, an optical isomer of sparteine. Optically active complexing agents can induce asymmetric carboxylation to yield products having about 10% to about 95% enantiomeric excess (e.e.) (see, e.g., Beak et al, j.org.chem., 1995, 60, 8148-. Equilibrating the trans-/cis-mixture in the presence of a suitable base to obtain a trans-predominant acid of formula 8, wherein the trans-/cis-ratio is 80: 20, 90: 10, 95: 5, or greater than 98: 2. Suitable bases include, for example, lithium hexamethyldisilazide, lithium diisopropylamide, or lithium 2, 2, 6, 6-tetramethylpiperidine.

In another embodiment, the carboxylic acid of formula 8 is obtained directly using 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] nonane as the complexing diamine, with trans-/cis-ratios of the isomers of 90: 10, 95: 5 or greater than 98: 2, avoiding an equilibration step.

A racemic mixture of the compound of formula 8 can be separated to provide a single enantiomer of formula 9. Known methods for the isolation of racemic amino acids may be used, including, but not limited to, crystallization of optically active amine salts, preparation of 2-carboxylic acid esters with optically active alcohols followed by crystallization or chromatographic separation, and preparation of optically active N-alkoxycarbonyl derivatives followed by crystallization or chromatography. In one embodiment, the (R) α -aminoethylbenzene or (S) 1-amino-1, 2, 3, 4-tetrahydronaphthalene salt of the compound of formula 8 is crystallized to yield the amine salt of formula 9.

The free acid of the salt of formula 9 obtained by extraction with, for example, aqueous sodium hydrogen sulfate is extracted with, for example, di-tert-butyl dicarbonate (Boc)₂O) esterification to give the ester of formula 10. Removal of the-COOR' protecting group under known conditions (e.g., methanesulfonic acid in an organic solvent such as t-butyl methyl ether or tetrahydrofuran) provides the compound of formula 1.

In another embodiment, bicyclic pyrrolidinyl compounds of formula 3 (as exemplified by compound 17 shown below) may be prepared generally according to scheme II.

Scheme II

Referring to scheme II, the camphorimine of formula 12 is prepared by reacting tert-butyl glycinate of formula 11 with (1S) - (-) camphor in the presence of a Lewis acid such as boron trifluoride etherate. The Michael addition of an amine of formula 12 to methyl cyclopentenecarboxylate gives an adduct of formula 13. The individual isomers of compound 13 shown are obtained by recrystallization of the crude product from a mixture of isopropanol and water. Removal of the camphorimine with hydroxylamine in the presence of sodium acetate followed by cyclization affords the lactam ester of formula 14. Optionally, the reaction mixture may be treated with succinic anhydride to facilitate recovery of the desired product of formula 14 and the camphor derivative of formula 15. The lactam of formula 14 is converted to its benzyloxycarbonyl derivative of formula 16 by treatment with a base such as sodium hydride followed by benzyl chloroformate. Reduction of the lactam of formula 16 with a hydride reducing agent such as borane-dimethylsulfide-piperidine provides the carbamate of formula 17. Removal of the benzyloxycarbonyl protecting group can be achieved under reducing conditions (such as hydrogen in the presence of a palladium catalyst such as palladium hydroxide) to give the desired bicyclic pyrrolidinylester of formula 17. Isolation of the ester of formula 17 is optionally achieved by formation of a salt, such as the oxalate salt of formula 1 a.

The invention further provides a preparation method of the compound of the formula 2. A specific example of a compound of formula 2, wherein R'₄Is H, R'₅Is n-propyl, R₄Is cyclopropyl, shown below as formula 18.

In one aspect, compound 18 can be prepared as outlined in scheme III.

Scheme III

In scheme III, the methoxymethylamide of formula 20 Cbz-norvaline is prepared by reacting Cbz-norvaline of formula 19 with methoxymethylamine in the presence of a coupling reagent such as EDC. Reduction of a compound of formula 20 with a hydride reagent such as lithium aluminum hydride or diisobutylaluminum hydride at a temperature of-20 ℃ to 10 ℃ provides the norvaline compound of formula 21. The preparation of the corresponding cyanohydrin of formula 22 is achieved by reacting a compound of formula 21 with an alkali metal cyanide, such as potassium cyanide, in the presence of an alkali metal thiosulfite, such as sodium thiosulfite. The compound of formula 22 is hydrolyzed in the presence of HCl in a suitable solvent such as dioxane at an elevated temperature of about 50 ℃ to 110 ℃ to produce the corresponding 3-amino-2-hydroxyhexanoic acid (not shown), which is converted to the Cbz derivative of formula 23 by reaction with Cbz-hydroxysuccinimide. The cyclopropyl amide of formula 24 is prepared from compound 23 by reaction with cyclopropylamine in the presence of a coupling reagent such as EDC. Removal of the Cbz group is achieved under known reducing conditions (such as hydrogen in the presence of a palladium catalyst) to give the compound of formula 18.

In another embodiment, the cyclopropylamide of formula 18 is prepared using a paselini (Passerini) reaction (see, e.g., a. doemling et al, angelw. chem., 2000, 112, 3300-.

Scheme IV

Referring to scheme IV, Cbz-valine 21 is reacted with a cyclopropylisocyanide of formula 25 (available from Oakwood Products, Inc., West Columbia, SC 29172, USA) in the presence of trifluoroacetic acid, optionally in the presence of an asymmetric catalyst to provide a cyclopropylamide of formula 24. See, e.g., Schreiber et al, org.lett.2004, 6, 4231. The intermediate trifluoroacetate (not shown) is hydrolyzed in isolation to afford compound 24 directly. Removal of the Cbz protecting group is achieved under reducing conditions as previously described to give the compound of formula 18.

In another embodiment, hydroxy-acid compounds of formula 23 can be prepared according to the methods described in U.S. patent nos. 6,020,518, 6,087,530, and 6,639,094, each of which is incorporated herein by reference in its entirety.

Although the methods shown in schemes III and IV above illustrate the synthesis of a particular compound (a compound of formula 18), the methods in schemes III and IV can be used to produce other compounds of formula 2.

In another embodiment, the invention further provides processes and intermediates for preparing compounds of formula 4, as illustrated in scheme V.

Scheme V

Referring to scheme V, bicyclic amino esters of formula 1 wherein R₃Is t-butyl, with a protected amino acid of formula 26 (wherein Z is an amine protecting group, which may be different from that used to remove R₃Removal of the protecting group under acidic, basic, or hydrogenation conditions) in the presence of a coupling reagent to provide an amide-ester of formula 27. Removing the protecting group Z from the amide-ester of formula 27 to provide an amine-ester compound of formula 28.

Reacting an amino group containing compound of formula 28 with a protected amino acid 29 in the presence of a coupling reagent gives a tripeptide of formula 30.

Removal of the protecting group Z from the tripeptide of formula 30 provides a free amino-tripeptide of formula 31.

Reacting the amino-tripeptide of formula 31 with pyrazine-2-carboxylic acid of formula 32 in the presence of a coupling reagent gives an amide-tripeptide ester of formula 33.

Hydrolysis of the ester of the amide-tripeptide ester of formula 33 provides the amido-tripeptide acid of formula 34.

Reacting the amido-tripeptide acid of formula 34 with an amino-hydroxyamide of formula 18 in the presence of a coupling reagent gives a hydroxy-peptide of formula 35.

In the final step, the hydroxy group of the compound of formula 35 is oxidized to provide the compound of formula 4.

Oxidation of compound 35 can be achieved with a variety of known oxidizing agents, such as: chromic acid in acetone; Dess-Martin oxidizer (Dess-Martin periodinane) (1, 1-dihydro-1, 1, 1-triacetoxy-1, 2-benziodoxazol-3 (1H) -one); sodium hypochlorite in the presence of TEMPO, and optionally an alkali metal halide such as sodium bromide.

In some embodiments, the hydroxyl configuration of 35 is a mixture of R and S isomers in a ratio of about 90: 10 to about 10: 90, typically in a ratio of about 60: 40 to about 40: 60.

In another embodiment, the hydroxy group of compound 35 has the R configuration with an enantiomeric excess of about 90% ee.

In a further embodiment, the hydroxy group of compound 35 has the S configuration with an enantiomeric excess of about 90% ee.

Any intermediate obtained as described herein may be used with or without isolation from the reaction mixture. Can be connected with proper RW-, P₂-、P₃-L₂-P₂Or P₄-L₃-P₃-L₂-P₂-partial derivatization of the desired protease inhibitor. The coupling of the amine to such moieties can be carried out using the corresponding carboxylic acid or reactive equivalent thereof under standard amide bonding or coupling conditions. Typical coupling reactions include a suitable solvent, an amine concentration ranging from about 0.01 to 10M, preferably from about 0.1 to about 4.0M, the necessary carboxylic acid, a base, and a peptide coupling reagent.

If the amine is used without isolation, the coupling can be carried out in situ in the solvent of the reaction mixture used to prepare the amine, or in a different solvent. The necessary carboxylic acid may be added to the reaction mixture to maintain the reactants at a temperature in the range of about 0 ℃ to 100 ℃, preferably about 20 ℃ to about 40 ℃. The base and peptide coupling reagent are then added to the mixture, which is maintained at a temperature in the range of about 0 ℃ to about 60 ℃, preferably about 20 ℃ to about 40 ℃. The base is typically a tertiary amine base such as triethylamine, diisopropylethylamine, N-methylmorpholine, DBU, DBN, N-methylimidazole, preferably triethylamine or diisopropylethylamine. The amount of base used is generally up to about 20 equivalents per equivalent of amine, preferably at least about 3 equivalents of base. Examples of peptide coupling reagents include DCC (dicyclohexylcarbodiimide), DIC (diisopropylcarbodiimide), di-p-toluoyl carbodiimide, BDP (1-benzotriazolediethylphosphate-1-cyclohexyl-3- (2-morpholinoethyl) carbodiimide), EDC (1- (3-dimethylaminopropyl) -3-ethyl-carbodiimide hydrochloride), cyanuric fluoride, cyanuric chloride, TFFH (tetramethylfluoromethamidine hexafluorophosphate), DPPA (diphenylphosphoryl azide), BOP (benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate), HBTU (O-benzotriazol-1-yl-N, N, N ', N' -tetramethyluronium hexafluorophosphate), TBTU (O-benzotriazol-1-yl-N, n, N ', N' -tetramethyluronium tetrafluoroborate), TSTU (O- (N-succinimidyl) -N, N, N ', N' -tetramethyluronium tetrafluoroborate), HATU (N- [ (dimethylamino) -1-H-1, 2, 3-triazolo [4, 5, 6] -pyridin-1-ylmethylene ] -N-methylmethanaminium hexafluorophosphate N-oxide), BOP-Cl (bis (2-oxo-3-oxazolidinyl) phosphinic acid chloride), PyBOP ((1-H-1, 2, 3-benzotriazol-1-yloxy) -tris (pyrrolidinyl) phosphonium tetrafluorophosphate), BrOP (bromotris (dimethylamino) phosphonium hexafluorophosphate), DEPBT (3- (diethoxyphosphoryloxy) -1, 2, 3-benzotriazin-4 (3H) -one) or PyBrOP (bromotris (pyrrolidinyl) phosphonium hexafluorophosphate). EDC, HOAT, BOP-Cl and PyBrOP are preferred peptide coupling reagents. The amount of peptide coupling reagent ranges from about 1.0 to about 10.0 equivalents. Optional reagents useful for the amide bond-forming reaction include DMAP (4-dimethylaminopyridine) or active ester reagents such as HOBT (1-hydroxybenzotriazole), HOAT (hydroxyazabenzotriazole), HOSu (hydroxysuccinimide), HONB (endo-N-hydroxy-5-norbornene-2, 3-dicarboxamide) in amounts ranging from about 1.0 to about 10.0 equivalents.

Alternatively, the amine may be substituted with a reactive equivalent of R₁Carboxylic acid treatment, e.g. RW-C (═ O) X¹、P₂-C(＝O)X¹、P₃-L₂-P₂-C(＝O)X¹Or P₄-L₃-P₃-L₂-P₂-C(＝O)X¹Wherein C (═ O) X¹Are groups that are more reactive than COOH in the coupling reaction. -C (═ O) X¹Examples of groups include wherein X¹Is a group of Cl, F, OC (═ O) R (R is, for example, aliphatic or aryl), -SH, -SR, -SAr or-SeAr.

Acids and amines known in the art for use hereinProtecting groups (see, e.g., t.w.greene)&M. G.M Wutz, "Protective Groups in Organic Synthesis," third edition, John Wiley&Sons, Inc (1999), and earlier versions of the book. Examples of suitable acid protecting groups include t-butoxy, benzyloxy, allyloxy, and methoxymethoxy. Examples of suitable amine protecting groups include methyl 9-fluorenylcarbamate, t-butyl carbamate, benzyl carbamate, trifluoroacetamide, and p-toluenesulfonamide. Rw-, P-compounds of various chemical groups known to be useful as protease inhibitors₂-、P₃-L₂-P₂Or P₄-L₃-P₃-L₂-P₂-a moiety. Examples of such groups are reported in the following publications: WO 97/43310, US 20020016294, WO 01/81325, WO 02/08198, WO 01/77113, WO 02/08187, WO 02/08256, WO02/08244, WO 03/006490, WO 01/74768, WO 99/50230, WO 98/17679, WO 02/48157, US 20020177725, WO 02/060926, US 20030008828, WO 02/48116, WO 01/64678, WO 01/07407, WO 98/46630, WO 00/59929, WO 99/07733, WO 00/09588, US 20020016442, WO 00/09543, WO 99/07734, US6,018,020, US6,265,380, US6,608,027, US 20020032175, US 20050080017, WO 98/22496, US5,866,684, WO 02/079234, WO 00/31129, WO 99/38888, WO 99/64442, WO 2004072243 and WO 02/18369, which is incorporated herein by reference in its entirety.

Although only a single stereoisomer of the compound of formula 4 is illustrated in scheme V, the present invention is intended to include all stereoisomers of formula 4 depicted in table I. All of these stereoisomers can be prepared in the same manner using reagents containing carbon atoms of different stereoconfigurations, e.g.

Example III

The following preparation examples are set forth in order to provide a more thorough understanding of the present invention. These examples are for illustrative purposes only and should not be construed as limiting the scope of the invention in any way.

Preparation 1: 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] nonane

Method 1

To a three-necked 5L flask equipped with a mechanical stirrer, thermocouple, condenser and addition funnel were added 1-propyl-4-piperidone (100g, 0.71mol), paraformaldehyde (50g, 1.67mol) and ethanol (2.0L) under nitrogen with stirring. Acetic acid (90mL, 1.56mol) was added and the mixture was warmed to 40 ℃. Propylamine (64mL, 0.78mol) was dissolved in ethanol (500mL) in a separate flask. This solution was added to the above mixture over 7-8 hours. The mixture was stirred at 40 ℃ for a further 1.5 hours and then cooled to ambient temperature. Passing the mixture through CelitePad filtration and rinsing of the Celite with ethanol (2X 100mL each). The solution was concentrated in vacuo and diethylene glycol (1.0L) was added. Potassium hydroxide (160g) was dissolved in water (190mL) in a separate flask. The solution was added to the diethylene glycol mixture while stirring, and then the mixture was warmed to 85 ℃. Hydrazine monohydrate (96mL) was added over 2 hours and the resulting mixture was stirred at 85 ℃ for an additional 1 hour. The mixture was warmed to a bath temperature of 160 ℃ under nitrogen, while the distillate was collected in a Dean-Stark trap. The lower aqueous phase was returned to the reaction flask while the upper product phase was collected. This process was repeated until the product no longer distilled as an azeotrope with water. During this process the tank temperature (pot temperature) changed from 135 to 160 ℃. The collected upper phase fractions were combined and dissolved in heptane (160 mL). The solution was washed with water (2 times 120mL each) and the combined aqueous phases were extracted with heptane (2 times 100mL each). The combined organic phases were concentrated to give the title compound.

¹H NMR(DMSO-d₆，500MHz)：δ2.60(dd，J＝10.88，2.04Hz，4H)，2.23(dd，J＝10.88，4.58Hz，4H)，2.12(t，J＝7.74Hz，4H)，1.91-1.84(m，2H)，1.44-1.35(m，6H)，0.85(t，J＝7.25Hz，6H)

Method 2

In a four-necked 12L flask equipped with a mechanical stirrer, a thermocouple and a condenser, acetic acid (260mL, 4.67mol) was added to a mixture of 1-propyl-4-piperidone (300g, 2.12mol), paraformaldehyde (150g, 5.00mol) and ethanol (6.00L) under a nitrogen atmosphere. The heterogeneous mixture was warmed to 40 ℃ and a solution of propylamine (192mL, 2.34mol) in ethanol (1.50L) was added over a period of 7.5 hours. After the addition was complete, the mixture was maintained at 40 ℃ for 1.5 hours. The mixture was cooled to 22-25 ℃ and filtered. The collected solids were washed with ethanol (2 times 200mL each) and the combined filtrates were concentrated to about 1.0L by vacuum distillation (90mmHg, 50-55 ℃). Diethylene glycol (2.60L) was added followed by a solution of potassium hydroxide (477g) in water (570 mL). The reaction mixture was heated to 85 ℃ and hydrazine monohydrate (279mL) was added over 2 hours. Heating was continued for 1 hour at 85 ℃ after the addition was complete, then the mixture was heated to 155 ℃ while the distillate was collected, which formed 2 layers. The lower layer was periodically returned to the reaction mixture. Heating was continued at 155 ℃ and 165 ℃ until distillation of the upper layer ceased. The upper product layer was diluted with heptane (480mL) and washed with water (2X, 240mL each). The combined aqueous phases were extracted with heptane (2X, 300mL each). The combined heptane extracts were concentrated to provide the title compound as a light yellow liquid.

Preparation 2: (S) -3-amino-N-cyclopropyl-2-hydroxyhexanamide (18)

A250 mL round bottom flask equipped with an overhead stirrer, addition funnel, thermocouple, and nitrogen/hydrogen inlet was purged with nitrogen for a few minutes. The protected amino-hydroxy acid (10.0g, 0.035mol) and N-hydroxysuccinimide (9.0g, 0.078mol, 2.2 molar equivalents) were added to the flask followed by 105mL of DMF. The mixture was stirred at 20 ± 5 ℃ until a clear solution was obtained (approximately 15 minutes). The flask was cooled to-9.8 ℃ (ice/acetone bath). EDC. HCl (13.6g, 0.071mol, 2.0 mol equivalent) was added in one portion to the flask. The flask contents were allowed to stir at-5. + -. 5 ℃ for 3 hours. The reaction flask contents were cooled to-10 ± 3 ℃ and cyclopropylamine (4.89g, 0.085mol, 2.4 molar equivalents) was added via the addition funnel while maintaining the temperature range at 5 ± 3 ℃. The reaction mixture was allowed to stir at 5 ± 5 ℃ for 60 minutes, then slowly warmed to room temperature and stirred overnight. The reaction mixture was transferred to a larger round bottom flask and quenched at room temperature by the addition of water (270 mL). The DMF/aqueous layer was extracted with 3 portions of 35-40 deg.C EtOAc (150mL), and the combined EtOAc extracts were washed with water (2X, 300mL each) followed by 10% NaHCO₃The solution (300mL) was washed and finally washed with water (300 mL). The EtOAc layer was concentrated at atmospheric pressure and heptane (100mL) was added. Continuing at 80The product was crystallized from solution by distillation at + -5 deg.C and addition of additional heptane (50 mL). The mixture was held at 85 ℃ for 2 hours, slowly cooled to room temperature, and held for 1 hour. The product was vacuum filtered and dried at 30 ℃ overnight at 25mmHg to give the crude product (12.86 g). An 11.44g portion of the crude product was placed in a 250mL round bottom flask, 50mL MTBE was added, and the thick slurry was stirred at room temperature for 3 hours. The product was filtered and the filter cake was washed with MTBE (50 mL). A sample of the dried product (6.4g) was taken for weight% determination (92.2 wt%) and HPLC A% (100A%).

A 1.0L step-wise (Buchi) hydrogenation vessel equipped with an overhead stirrer, ballast tank, thermocouple and nitrogen/hydrogen inlet was purged with nitrogen for several minutes. The protected amino-hydroxyamide (49.9g, 0.156mol, prepared as described above) was reacted with 20% Pd (OH)₂Carbon (2.85g, 0.002mol, 50% by weight water) was added to the vessel followed by 700mL of MeOH. The mixture was stirred at 40 ℃ until the starting material dissolved (approximately 15 minutes). The vessel and ballast tank were purged 2 times to 40psig with nitrogen, vented to atmospheric pressure with nitrogen, and pressurized to 40psig 2 times with hydrogen, each time vented to atmosphere. The ballast tank is finally pressurized to 400psig and the vessel is pressurized to 30psig via the ballast tank. The hydrogenation vessel was maintained at 40 ℃ and 30psig hydrogen (conditioned via ballast tanks) for 2 hours. The vessel was vented to atmospheric pressure with nitrogen and the slurry was sampled for HPLC analysis of the residual starting material (1.8%; both diastereomer limits 0.5%). The vessel was re-purged with hydrogen and re-pressurized to 30psig and held at 40 ℃ for an additional 30 minutes. The vessel was vented to atmospheric pressure with nitrogen and a sample of the slurry was analyzed by HPLC for residual amino-amide (1.1%; both diastereomer limits 0.5%). The vessel was re-purged with hydrogen and re-pressurized and held at 40 ℃ for an additional 40 minutes. The vessel was vented to atmospheric pressure and kept under a nitrogen atmosphere overnight.

HPLC analysis of the remaining protected amino-hydroxyamide was performed on the sample (no detection; both diastereomer limits ≦ 0.5%). During stirring overnight a portion of the product crystallized out of solution and another 300mL of MeOH was added to dissolve the product. The slurry was warmed to 45 ℃ to ensure dissolution and then passed through Celite at 45 ℃And (4) bed filtration. The wet cake was rinsed with MeOH (250mL) and the filtrate was distilled to a volume of about 150mL at atmospheric pressure. Ethyl acetate (300mL) was added and distillation continued at atmospheric pressure, again to a volume of 150 mL. The procedure was repeated 2 more times. Heptane (150mL) was added to the 75 ℃ flask and the contents allowed to cool to room temperature and finally to 5 ℃ in an ice/water bath. The crystallized product was collected and the wet cake was washed with heptane (75mL) and dried overnight at 40 ℃ under reduced pressure. The free amino-amide was isolated as an off-white solid (21.2g, 0.114mol, 73.1% yield) with an HPLC purity of 98.5A% and a weight/weight determination of 94.2 wt%.

Example 1: n-tert-butoxycarbonyl-3-azabicyclo [3.3.0] octane (6)

Method 1

To a 2L 3-necked round bottom flask equipped with a mechanical stirrer, 500mL addition funnel, and thermometer was added 3-azabicyclo [3.3.0] under nitrogen]Nonane hydrochloride (100g, 0.677mol), potassium carbonate (187g, 1.35mol), tert-butyl methyl ether (220mL) and water (160mL) with stirring. The mixture was cooled to 14-16 ℃. Boc was added to a separate 500mL Erlenmeyer flask₂O (di-tert-butyl dicarbonate) (145g, 0.644mol) and tert-butyl methyl ether (190 mL). The mixture was stirred until complete dissolution was obtained. The solution was poured into an addition funnel and added to the reaction mixture above, maintaining the reaction temperature below 25 ℃. Water (290mL) was added to dissolve the solids and the mixture was stirred for 10-15 minutes. After separation of the lower aqueous phase, the organic phase was treated with 5% aq₄(2 washes, 145mL each) followed by water (145 mL). The organic phase was concentrated and methyl tert-butyl ether (1.3L) was added to give a solution of the title compound in tert-butyl methyl ether. See, e.g., r.griot, helv.chim.acta, 42, 67 (1959).

Method 2

Potassium carbonate (187g, 1.35mol) in water (160mL) was added to 3-azabicyclo [3.3.0]Octane hydrochloride (100g, 0.677mol) and tert-butyl methyl ether (220mL) and the resulting mixture was cooled to 14-16 ℃. Addition of Boc₂A solution of O (145g, 0.644mol) in tert-butyl methyl ether (190mL) was maintained at a temperature below 35 ℃. After the addition, the mixture was stirred for 1 hour and then filtered. The solid was washed with MTBE (50 mL). The phases were separated and the organic phase was treated with 5% aq₄(2 times, 145mL each) and water (145mL) and concentrated to 300mL in vacuo. MTBE (300mL) was added and the mixture was concentrated to remove water to less than 550 ppm. The concentrate was diluted with MTBE (400mL) to provide a solution of the title compound in MTBE.

Example 2: rac-2- (tert-Butoxycarbonyl) octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid (7)

Method 1

The solution of example 1, method 1 was charged to a 5L 4-necked flask equipped with a mechanical stirrer, addition funnel, ReactIR probe, and thermometer. Reacting 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1]]Nonane (183g, 0.88mol) was added to the flask. Data collection was started on the ReactIR instrument and the solution was allowed to cool to-72 to-75 ℃. Sec-butyllithium (600mL, 1.6M in cyclohexane) was slowly added to the reaction mixture, maintaining the reaction temperature below-69 ℃. The addition was monitored with a ReactIR instrument at 1698cm for 3 consecutive scans (2 min intervals)^-1Absorbance had disappeared and 1654cm^-1The addition was stopped after the absorbance stopped increasing. The solution was stirred at-75 to-72 ℃ for 3 hours. Introducing CO₂A10% mixture under nitrogen was carefully charged into the reaction mixture, maintaining the reaction temperature below-70 ℃. In CO₂The absorbance appears in the ReactIR spectrum (2350 cm)^-1) And then stopping the inflation. Heating the mixture to 0-5 deg.C, adding30% by weight NaHSO₄Solution (1.4L). The mixture was warmed to 22-25 ℃ and stirred for 30 minutes. The aqueous phase was separated and the organic phase was washed with water (700 mL). The aqueous phase was decanted and the organic phase was concentrated to provide the title compound.

Method 2

In a flask equipped with mechanical stirrer, addition funnel, ReactIR probe and thermometer, 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1]A solution of nonane (183g, 0.87mol) in MTBE (300mL) was added N-tert-butoxycarbonyl-3-azabicyclo [3.3.0] example 1, method 2]The mixture was cooled to-75 to-72 ℃ in a solution of octane. Sec-butyllithium solution (510mL, 1.6M) was added, maintaining the reaction temperature below-70 ℃ until 1698cm^-1Absorbance had disappeared and 1654cm^-1The absorbance stopped increasing. The solution was stirred at-75 to-72 ℃ for 3 hours. The reaction mixture is treated with N₂10% CO in₂Aerating and keeping the reaction temperature below-70 ℃. When CO is present₂The absorbance appears in the ReactIR spectrum (2339 cm)^-1) The inflation is stopped. Heating the mixture to 0-5 deg.C, adding 30 wt% NaHSO₄Solution (1.4L), the mixture was warmed to 22-25 ℃ and then stirred for 30 minutes. The phases were separated and the aqueous phase was checked to ensure the pH was below 3. The organic phase was washed with water (700mL) and then concentrated to 300 mL. Ethyl acetate (1.7L) was added and the mixture was concentrated to 300mL 2 times to give a solution of the title compound in ethyl acetate.

Example 3: (1S, 3aR, 6aS) -2- (tert-Butoxycarbonyl) octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid (S) -1, 2, 3, 4-tetrahydronaphthalene-1-ammonium (9a)

Method 1

Ethyl acetate (2.3L) was added to the residue of method 1 of example 2 using CeliteThe pad filters the mixture. (S) -1, 2, 3, 4-tetrahydro-1-naphthylamine (56.7g, 0.385mol) was added, and the solution was stirred at 22-25 ℃ for 3-4 hours. The mixture was filtered and the solid was rinsed with ethyl acetate (200 mL). The solid was dried under vacuum at 20-30 ℃ for 4 hours to give the title compound.

To a 3-neck RBF equipped with a temperature controller, mechanical stirrer, reflux condenser and nitrogen bubbler was added (S) -1, 2, 3, 4-tetrahydro-1-naphthylammonium salt (88.98g, 0.22mol), ethyl acetate (712mL) and 2-propanol (666 mL). The mixture was warmed to 70-75 ℃ with stirring. The mixture was stirred for 15-30 minutes and then cooled to-5 to-10 ℃ over 1 hour. The resulting slurry was filtered and the solid was rinsed with cold ethyl acetate (180mL) to give the title compound as a white solid.

Method 2

An ethyl acetate solution of racemic N-tert-butoxycarbonyl-3-azabicyclo [3.3.0] octane-2-carboxylic acid, example 2, method 2, was added to a solution of (S) -1, 2, 3, 4-tetrahydro-1-naphthylamine (56.7g, 0.385mol) in ethyl acetate (300 mL). The mixture was stirred at 22-25 ℃ for 3-4 hours, then filtered and the solid washed with ethyl acetate (200 mL). The product was dried under vacuum at 20-30 ℃ for 4 hours to give the title compound.

A mixture of the salt (89.0g) prepared as above, ethyl acetate and 2-propanol was warmed to 70-75 ℃ until complete dissolution. The mixture was cooled to-5 to-10 ℃ over 2 hours and stirred for 3-4 hours. The mixture was filtered and the product was dried at 35-40 ℃ to give the title compound.

Example 4: (R) -1-Phenylethylammonium (1S, 3aR, 6aS) -2- (tert-Butoxycarbonyl) octahydrocyclopenta [ c ] pyrrole-1-carboxylate (9b)

To a solution of racemic N-tert-butoxycarbonyl-3-azabicyclo [3.3.0] octane-2-carboxylic acid (4.66g) in ethyl acetate (100mL) was added (R) - α -methylbenzylamine (56.7g), and the solution was stirred at 22-25 ℃ for 16 hours. The mixture was filtered and the solid was rinsed with ethyl acetate. The solid was dried under vacuum at 20-30 ℃ for 4 hours to give the title compound.

Example 5: (1S, 3aR, 6aS) -tert-butyl octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid tert-butyl ester oxalate

Method 1

(S) -1, 2, 3, 4-tetrahydro-1-naphthylammonium salt (81.7g, 0.203mol), prepared according to method 1 of example 3, tert-butyl methyl ether (400mL) and 5% NaHSO₄-H₂A mixture of O (867mL, 0.304mol) was stirred for 30 minutes until all the solid dissolved. The organic phase was washed with water (334mL) and then concentrated to 259 mL. T-butyl methyl ether (334mL) was added and the solution was reconcentrated to 259 mL. The addition-concentration process was repeated 2 more times. After final concentration, t-BuOH (158mL) and dimethylaminopyridine (5.04g, 41.3mmol) were added. Addition of Boc₂A solution of O (67.6g, 0.31mol) in t-butyl methyl ether (52.0 mL). After stirring at ambient temperature for 5 hours, tert-butyl methyl ether (158mL) and 5% aqueous NaHSO were added₄-H₂O (260mL), and the resulting mixture was stirred. The organic phase was washed with 5% aqueous NaCl (2X, 260mL each). The organic phase was concentrated to 320mL and tetrahydrofuran (320mL) was added. The organic phase was again concentrated to 320mL and tetrahydrofuran (320mL) was added. After further concentration to 320mL, methanesulfonic acid (80.1g, 0.62mol) was added and the solution was stirred at ambient temperature for 4.5 h. Adding the reaction mixture to K₂CO₃30% aqueous solution (571mL) and stirred. The aqueous phase was extracted with isopropyl acetate (320 mL). The combined organic phases were concentrated to 320mL and isopropyl acetate (320mL) was added. The organic solution was further concentrated to 320 mL. The organic phase was washed with water (320 mL). Isopropyl acetate (320mL) was added to the organic phase and the solution was concentrated to 192 mL.A second addition of isopropyl acetate (320mL) concentrated the organic solution to 192 mL. A solution of oxalic acid (24.1g, 267mmol) in isopropyl acetate (448mL) was added to the organic solution over 2 hours. The mixture was stirred for 2-4 hours and the slurry was filtered. The white solid was rinsed with isopropyl acetate (100mL) and dried under vacuum at 35-40 ℃ to give the title compound.

Method 2

(S) -1, 2, 3, 4-tetrahydro-1-naphthylammonium salt (148g, 0.609mol), tert-butyl methyl ether (726mL), prepared according to the method of example 3, method 2 and 5% NaHSO were stirred₄-H₂A mixture of O (1.58L, 0.913mol) until all solids were dissolved. The phases were separated and the organic phase was washed with water (726 mL). The organic phase was concentrated to about 400 mL. Tert-butyl methyl ether (726mL) was added and the mixture was concentrated to 590 mL. The addition of tert-butyl methyl ether and concentration was repeated to give a final volume of 350 mL. Dimethylaminopyridine (8.42g, 68.9mmol) and tert-butanol (260mL) were added followed by Boc addition over 0.5 h₂A solution of O (112g, 0.52mol) in MTBE (88 mL). The mixture was stirred at 22-25 ℃ for 5 hours. A solution of 5% sodium bisulfate in water was added and the mixture was stirred for 0.5 hour. The organic phase was washed with 5% sodium chloride (2 times 440mL each) and concentrated to 270 mL. Tetrahydrofuran (540mL) was added and the mixture was concentrated to 270 mL; this procedure was repeated 2 more times to give a final volume of 270 mL. Methanesulfonic acid (67mL) was added over 0.5 hour while maintaining the temperature below 30 deg.C and the mixture was stirred at 22-25 deg.C for 12 hours. The mixture was added to a 30% aqueous solution of potassium carbonate (478mL) while maintaining the temperature at 22-25 ℃. The mixture was filtered, the phases separated and the aqueous phase was extracted with isopropyl acetate (2 times 540mL each). The organic phase was concentrated to 270mL and then evaporated 2 times with isopropyl acetate (540mL) to give a final volume of 540 mL. The organic phase was washed with water (2 times 540mL) and then evaporated 2 times with isopropyl acetate (320mL) to give a final volume of 320 mL. Additional isopropyl acetate (429mL) was added followed by a solution of oxalic acid (40.4g, 0.448mol) in t-butyl methyl ether (321mL) over 2 hours, maintaining a temperature of 22-25 ℃. The mixture was stirred at 22-25 ℃ for 3 hours and then filtered. The filter cake was washed with isopropyl acetate (100mL) and the product was dried at 35-40 deg.CAir dried to give the title compound as a white solid.

Example 6: (1S, 3aR, 6aS) -2- ((S) -2- (benzyloxycarbonylamino) -3, 3-dimethylbutyryl) octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid tert-butyl ester (27)

Method 1

A3-L3-necked round bottom flask equipped with an overhead stirrer, condenser, thermocouple, and nitrogen outlet was purged with nitrogen for several minutes. In a separate flask, sulfuric acid (46.2mL, 0.867mol) was diluted with 442mL of water. The solution was allowed to cool slightly. Cbz-L-tert-leucine dicyclohexylamine salt (330.0g, 0.739mol) was added to the reaction flask. Tert-butyl methyl ether (1620mL) was added to the reactor and the mixture was stirred to suspend the salt. The acid solution prepared above was added to the reactor over about 10 minutes, maintaining the temperature at 20 ± 5 ℃. The mixture was stirred at room temperature for about 1 hour, then slowly diluted with water (455 mL). Agitation was stopped and the layer was allowed to settle. The lower (aqueous) phase was removed to provide 1100mL of a colorless solution at pH 1. Additional water (200mL) was added to the organic phase remaining in the flask. The mixture was stirred at room temperature for about 1 hour. Agitation was stopped and the layer was allowed to settle. The lower (aqueous) phase was removed to provide 500mL of a colorless solution at pH 2. The organic phase was heated to about 35 ℃, diluted with DMF (300mL), and concentrated under reduced pressure to a point where distillation was significantly slowed, leaving about 500mL of concentrate. The concentrate was transferred to a 1-L Schott bottle without rinsing. The concentrate (clear colorless solution) weighed 511.6 g. Based on the solution assay and the weight of the solution, the solution contained 187.2g (0.706mol) of Cbz-L-tert-leucine.

To a 5-L4-necked round bottom flask equipped with an overhead stirrer, thermocouple, addition funnel, and nitrogen inlet was added HOBT. H₂O (103.73g, 0.678mol, 1.20 mol equivalent), EDC & HCl (129.48g, 0.675mol, 1.20 mol equivalent), and DMF (480 mL). The slurry was cooled to 0-5 ℃. Warp beamA36.6% by weight solution of the acid of Cbz-L-tert-leucine in DMF (491.3g, 0.745mol, 1.32 molar equivalents) was added to the reaction mixture for 47 minutes while maintaining the temperature at 0-5 ℃. The reaction mixture was stirred for 1 hour 27 minutes. A solution of 3-azabicyclo (3.3.0) octane-2-carboxylic acid tert-butyl ester in isopropyl acetate (28.8% by weight, 414.3g, 0.564mol) was added over 53 minutes while maintaining the reaction temperature at 0-5.1 ℃. The reaction mixture was warmed to 20 ± 5 ℃ over about 1 hour. 4-methylmorpholine (34.29g, 0.339mol, 0.60 mol equiv) was added over 5 minutes. The reaction mixture was stirred for 16 hours, then isopropyl acetate (980mL) was added to the reaction solution. A solution of histamine-2 HCl (41.58g, 0.226mol, 0.40 molar equivalents) in water (53.02g) was added to the reaction mixture over 4 minutes followed by 4-methylmorpholine (45.69g, 0.45mol, 0.80 molar equivalents). The reaction mixture was sampled after 3.5 hours. Water (758mL) was added and the mixture was stirred for about 20 minutes and then allowed to settle for 11 minutes. The phases were separated. The aqueous phase was extracted with isopropyl acetate (716mL) and the organic phases were combined. 1N aq. HCl was prepared by adding 37 wt% hydrochloric acid (128.3mL) to water (1435 mL). The organic phase is washed with 1N hydrochloric acid for about 20 minutes. By making K₂CO₃(171g, 1.23mol, 2.19 mol equiv.) in water (1540mL) to prepare 10 wt.% aq.K₂CO₃And (3) solution. The organic phase is treated with 10% by weight aq₂CO₃The solution was washed for about 20 minutes. The final clear, very slightly yellow organic solution (weighing 1862.1g) was sampled and subjected to solution testing. The solution contained 238.3g (0.520mol) of the title compound product, based on solution determination and solution weight.

¹H NMR(DMSO-d₆，500MHz)：δ7.37ppm(5H，s)，7.25-7.33ppm(1H，m)，5.03ppm(2H，s)，4.17ppm(1H，d)，3.98ppm(1H，d)，3.67-3.75ppm(2H，m)，2.62-2.74ppm(1H，m)，2.48-2.56ppm(1H，m)，1.72-1.89ppm(2H，m)，1.60-1.69ppm(1H，m)，1.45-1.58ppm(2H，m)，1.38ppm(9H，s)，1.36-1.42ppm(1H，m)，0.97ppm(9H，s).

Method 2

A solution of potassium carbonate (73.3g) in water (220mL) was added to a suspension of tert-butyl (1S, 2S, 5R) 3-azabicyclo [3.3.0] octane-2-carboxylate oxalate (80.0g) in isopropyl acetate (400mL) while maintaining a temperature of about 20 ℃. The mixture was stirred for 0.5 h, the phases were separated and the organic phase was washed with 25% w/w aqueous potassium carbonate (80mL) to give a solution of the free base. In a separate flask, aqueous sulfuric acid (400mL, 0.863M) was added to a suspension of Cbz-tert-leucine dicyclohexylamine salt (118.4g) in tert-butyl methyl ether (640mL) while maintaining a temperature of about 20 ℃. The mixture was stirred for 0.5 h, the phases were separated and the organic phase was washed with water (200 mL). The phases were separated, N-methylmorpholine (80mL) was added to the organic phase, which was concentrated to 80mL at 40 ℃ under reduced pressure to give the free acid as a solution in N-methylmorpholine. The solution was added to a mixture of EDC & HCl (50.8g) HOBt hydrate (40.6g) in N-methylmorpholine (280mL) at 0-10 ℃. The mixture was stirred at about 5 ℃ for 1 hour. A solution of the above tert-butyl 3-azabicyclo [3.3.0] octane-2-carboxylate was added at 0-20 ℃ followed by N-methylmorpholine (32 mL). The mixture was stirred for 6 hours, then diluted with isopropyl acetate (600mL) followed by 1N HCl (400 mL). After stirring for 0.5 h, the phases were separated and the organic phase was washed with 25% w/w aqueous potassium carbonate (400mL) and water (80 mL). The mixture was stirred for about 1 hour and the phases separated to give a solution of the title compound in isopropyl acetate.

Method 3

(1S, 2S, 5R) 3-azabicyclo [3.3.0] octane-2-carboxylic acid tert-butyl ester oxalate (1.0 equiv.) was suspended in isopropyl acetate (6 vol.) at 20-25 deg.C, and potassium carbonate (3.0 equiv.) in water (3.5 vol.) was added. The mixture was stirred for 3 hours, then the phases were separated. The organic phase was washed with water (2 volumes).

Cbz-tert-leucine dicyclohexylamine salt (1.05 eq) was suspended in isopropyl acetate (6 vol) at 20-25 ℃ and sulfuric acid (1.3 eq) in water (5 vol) was added. The mixture was stirred for 30 minutes, the phases were separated and the organic phase was washed 2 times with water (2.5 volumes each).

The 2 solutions were combined and then cooled to 0-5 ℃. HOBt hydrate (1.1 equiv.) and EDC (1.1 equiv.) were suspended in the mixture and the mixture was stirred for 6 hours. The mixture was washed with water (5 volumes) and the resulting organic phase was treated with L-lysine (1 equivalent) and N-methylmorpholine (NMM) (2 equivalents) at 20-25 ℃ to destroy excess activated ester. The mixture was then washed with 5% potassium carbonate (5 vol), 1N hydrochloric acid (5 vol), 5% potassium carbonate (5 vol) and 2 times water (5 vol each) to give a solution of the title compound in isopropyl acetate.

Example 7: (1S, 3aR, 6aS) -2- ((S) -2-amino-3, 3-dimethylbutyryl) -octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid tert-butyl ester (28)

Method 1

The 1L staged hydrogenator was purged 3 times with nitrogen. Reacting (1S, 3aR, 6aS) -2- ((S) -2- (benzyloxycarbonylamino) -3, 3-dimethylbutyryl) octahydrocyclopenta [ c)]A12.8 wt.% solution of tert-butyl pyrrole-1-carboxylate (prepared by the method of example 6, method 1) in isopropyl acetate (39.39g, 0.086mol) was added in 307.8g portions to the reactor. Isopropyl acetate (100mL) was added to the reactor. Preparation of 50% Water and Wet 20% Pd (OH)₂Carbon (3.97g) in isopropyl acetate (168mL) was added to the reactor and agitation was started. The reactor was pressurized to 30psig with nitrogen and vented down to atmospheric pressure. This was repeated 2 times. The reactor was pressurized to 30psig with hydrogen and vented down to atmospheric pressure. This was repeated 2 times. The reactor was pressurized to 30psig with hydrogen and stirred at ambient temperature for 1 hour. The mixture was filtered through a buchner funnel with Whatman #1 filter paper to remove the catalyst. The filter cake was washed with isopropyl acetate (80 mL). This procedure was repeated 2 more times with 617g and 290.6g of a 12.8 wt% solution of the starting Cbz compound. The 3 times hydrogenated material was combined and distilled under reduced pressure (28 "Hg). The obtained solution (468.68g) was assayed for the title compound.

¹H NMR(DMSO-d₆，500MHz)：δ3.96ppm(1H，d)，3.67ppm(1H，dd)，3.53ppm(1H，dd)，3.19ppm(1H，s)，2.66-2.75ppm(1H，m)，2.49-2.53ppm(1H，m)，1.75-1.92ppm(2H，m)，1.66-1.74ppm(1H，m)，1.48-1.60ppm(4H，m)，1.38ppm(9H，s)，1.36-1.42ppm(1H，m)，0.91ppm(9H，s)

Method 2

The Cbz derivative 27 solution from example 6, method 2, was charged to 20% Pd (OH) in a hydrogenation unit₂Water (50%, 12.2 g). The apparatus was pressurized to 30psi with hydrogen and then stirred at about 20 ℃ for 2 hours. The mixture was filtered to remove the catalyst and the filter cake was washed with isopropyl acetate (160 mL). The combined filtrates were evaporated 2-3 times with about 4 volumes of heptane at 40 ℃ to remove isopropyl acetate. The resulting slurry was cooled to 0 ℃, filtered, and the product was dried in vacuo to give the title compound.

Method 3

(1S, 3aR, 6aS) -2- ((S) -2-amino-3, 3-dimethylbutyryl) -octahydrocyclopenta [ c ] of method 3 of example 6]Solution of pyrrole-1-carboxylic acid tert-butyl ester in isopropyl acetate 20% Pd (OH)₂(2% by weight load, 50% wet) the mixture was hydrogenated at 2bar and 20-25 ℃ for 2 hours. The catalyst was removed by filtration and washed with isopropyl acetate (2 volumes). The filtrate was concentrated to 10 volumes at 40 ℃ under reduced pressure to give a solution of the title compound in isopropyl acetate.

Example 8: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2- (benzyloxycarbonylamino) -2-cyclohexylacetylamino) -3, 3-dimethylbutyryl) octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid tert-butyl ester (30)

Method 1

To a 3L 3-necked round bottom equipped with an overhead stirrer, thermocouple, addition funnel, nitrogen outlet and ice/water bathHOBt & H is added into the flask₂O (51.74 g; 0.338mol, 1.05 molar equivalents), EDC & HCl (64.8 g; 0.338mol, 1.05 molar equivalents), followed by DMF (197.1g, 208.8mL) was added and agitation was started. The slurry was cooled to 0-5 deg.C and then a solution of acid 29(98.45 g; 0.338mol, 1.05 molar equivalents) in DMF (172.4 g; 182.9mL) was prepared and charged to the addition funnel. It was added to the batch over about 30 minutes, maintaining the temperature at 0-5 ℃. Once the addition was complete the reaction mixture was stirred at 0-5 ℃ for 2 hours. A solution of amine 28 in isopropyl acetate (450g solution; containing 104.4g of acid 29, 0.322mol) was added to the addition funnel and added dropwise over 1 hour, maintaining the temperature at 0-5 ℃. Sample analysis indicated incomplete reaction and additional EDC hydrochloride (3.89g) was added. After 3 hours, sample analysis showed 1.8% amine 28 remaining. Preparation of HOBT. H₂A slurry of O (2.59 g; 0.0169mol) and EDC & HCl (3.24 g; 0.0169mol) in DMF (10.44mL) was cooled to 0-5 ℃. A solution of acid 29(4.92 g; 0.169mol) in DMF (10.44mL) was prepared and EDC. HCl and HOBT in DMF were added over 30 minutes, maintaining the reaction temperature at 0-5 ℃. The mixture was stirred at 0-5 ℃ for 1 hour and then added to the original mixture maintained at 0-5 ℃. The mixture was stirred at about 25 ℃ for 14 hours. A solution of histamine-2 HCl (11.84 g; 0.064mol) in water (8.9mL) was prepared and added to the reaction mixture over 5-10 minutes. 4-methylmorpholine (13.01 g; 0.129mol) was fed into the batch over about 10 minutes, maintaining the batch temperature at 20. + -. 5 ℃. The reaction mixture was diluted with isopropyl acetate (443mL) then water (585 mL). A solution of potassium carbonate (57.8g) in water (585mL) was added and the mixture was stirred for 0.5 h. The layers were separated and the aqueous layer was extracted 2 times with isopropyl acetate (2 times 235mL each). The combined organic phases were washed with 18% aqueous HCl in water (585mL) followed by NaHCO in water (585mL)₃(43.25g) washing. The layers were separated to give a pale yellow solution of product 30 in isopropyl acetate weighing 1159.3g (1275mL) containing 16.0 w/w% 30 in isopropyl acetate.

¹H NMR(DMSO-d₆，500MHz)：δ7.74(1H，d)，7.36(5H，m)，7.34-7.26(1H，m)，5.01(2H，s)，4.51(1H，d)，4.02(1H，t)，3.96(1H，d)，3.73(1H，m)，3.66(1H，m)，3.68(1H，m)，2.53(1H，m)，1.86-1.76(2H，m)，1.70-1.30(10H，m)，1.39(9H，s)，1.15-0.85(5H，m)，0.96(9H，s).

Method 2

A solution of Cbz acid 29(59.62g) in N-methylpyrrolidone (126mL) was added to a suspension of EDC.HCL (39.23g) HOBt hydrate (31.34g) in N-methylpyrrolidone (221mL) while maintaining the temperature at about 0 ℃. After addition, the mixture was stirred at about 0 ℃ for 1.5 hours. A solution of amine 28(63.24g, prepared according to example 7, method 2) in isopropyl acetate (632mL) was added to the mixture, maintaining the temperature at about 0 ℃. After addition the mixture was allowed to warm to room temperature and stirred for 5 hours. A solution of potassium carbonate (20.17g) in water (316mL) was added while maintaining the temperature at about 20 ℃. The mixture was stirred vigorously for 0.5 hour. The phases were separated and the organic phase was vigorously stirred with potassium carbonate (105.3g) in water (316 mL). The organic phase was separated, washed with 1N HCl (316mL) and then water (158mL) to give the title compound 30 as a 12.7% w/w solution in isopropyl acetate.

Method 3

To a solution of (1S, 3aR, 6aS) -2- ((S) -2-amino-3, 3-dimethylbutyryl) -octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid tert-butyl ester (1 eq) in isopropyl acetate (10 vol) was added NMP (5 vol), followed by EDC (1.15 eq), HOBT hydrate (1.0 eq) and (S) -2- (benzyloxycarbonylamino) -2-cyclohexylacetic acid (29, 1.05 eq) and the suspension was stirred at 20-25 ℃ for 4 h. The mixture was washed with 5% potassium carbonate (5 vol). A mixture of glycine (1 eq), NMM (2 eq) and water (1 vol) was added and the mixture was stirred for 4 hours. The mixture was then washed with 5% potassium carbonate (5 vol), 1N hydrochloric acid (5 vol), 5% potassium carbonate (5 vol) and 2 times water (5 vol each) to give a solution of the title compound in isopropyl acetate.

Example 9: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2-amino-2-cyclohexylacetylamino) -3, 3-dimethylbutanoyl) octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid tert-butyl ester (31)

Method 1

A60-gallon Hastelloy (Hastelloy) hydrogenation reactor was charged with a solution of Cbz peptide 30(15.1kg) in isopropyl acetate (109 kg). The solution was reduced to 68L at 50 ℃ under vacuum. The mixture was then cooled to 25. + -. 5 ℃ and MeOH (15.4kg) was added. The mixture was flowed into a vessel and the reactor was dried. Adding Pd (OH) into the dry reactor₂C (20%, 1.51 kg). The solution containing the Cbz peptide 30 was added to the reactor and covered with hydrogen (30 psi). The reaction was stirred at 150-. After completion, a slurry of activated carbon (0.97kg) in isopropyl acetate (6.8kg) was added in portions and the mixture was stirred for 15 minutes. Passing the mixture through Celite(2.0kg) was filtered through a Sparkler filter and through a 0.1-um cartridge filter. The reactor was rinsed with isopropyl acetate (33.0kg) and the rinse was combined with the reaction mixture. The system was additionally rinsed with a mixture of isopropyl acetate (25.6kg) and MeOH (5.73 kg). The combined organics were reduced to 30L under vacuum at 65 ℃. The solution was cooled to 20-30 ℃ and heptane (30.8kg) was added. Distillation was started again and the mixture was reduced to 30L. The procedure was repeated for a total of 4 additions of heptane (above) and solvent reductions (above). The mixture was cooled to 0-5 ℃, the product was filtered and washed with heptane (12.6 kg). The wet solid (14.0kg) was dried under vacuum at 15-20 ℃ to constant weight to give the title compound.

¹H NMR(DMSO-d₆，500MHz)：δ7.97(1H，d)，4.49(1H，d)，3.96(1H，d)，3.76(1H，m)，3.67(1H，m)，3.05(1H，d)，2.70(1H，m)，2.53(1H，m)，1.87-1.77(2H，m)，1.7-1.3(10H，m)，1.39(9H，s)，1.2-0.85(5H，m)，0.96(9H，s).

Method 2

A solution of Compound 30 from method 1, example 8 was charged to 50% wet 20 wt% Pd (OH) in a pressure reactor₂Carbon (3.16 g). The reactor was pressurized with hydrogen at 30psi and the mixture was stirred for about 1 hour. The catalyst was filtered, the filter was washed with isopropyl acetate, and the combined organics were distilled to about 65 mL. The mixture was evaporated several times with heptane (316mL) until analysis indicated < 0.5% isopropyl acetate. The resulting slurry was diluted to 320mL and then warmed to reflux. The solution was allowed to cool slowly to about 5 ℃, the suspension was stirred for 1 hour and then filtered. The filter cake was washed with about 65mL heptane and the product was dried under vacuum at 30 ℃ to give the title compound as a white solid.

Method 3

(1S, 3aR, 6aS) -2- ((S) -2- ((S) -2- ((S) -2- (benzyloxycarbonylamino) -2-cyclohexylacetylamino) -3, 3-dimethylbutyryl) octahydrocyclopenta [ c ] of method 3 of example 9]A solution of tert-butyl pyrrole-1-carboxylate in isopropyl acetate, 20% Pd (OH) added₂(2% by weight load, 50% wet) the mixture was hydrogenated at 2bar and 20-25 ℃ for 2 hours. The catalyst was removed by filtration and washed with isopropyl acetate (1 vol). The solvent was exchanged by 2 times reflux distillation with heptane (8.6 vol). The mixture was cooled to 78 ℃ over 1 hour and then to 22 ℃ over 2 hours. After 1 hour at 22 ℃ the suspension was filtered, the filter cake was washed with heptane (3.2 volumes) and the product was dried under vacuum at 30 ℃ with a nitrogen purge to give the title compound.

Example 10: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid tert-butyl ester (33)

Method 1

To a 100mL round bottom flask was added pyrazine-2-carboxylic acid 32(1.6070g, 12.95mmol) and DMF (4 mL). The slurry was stirred at 20-25 ℃. Meanwhile, a CDI solution was prepared by combining CDI (2.1012g, 12.96mmol, 1 molar equivalent) and DMF (8.80g, 9.3mL) in a 25mL flask. Mild heating (30 ℃) helped dissolve. The CDI solution was cooled to 20-25 ℃ and added to the slurry of pyrazine-2-carboxylic acid. Stirring was continued for 1.5 hours to ensure complete activation of the acid, while carbon dioxide was produced as a by-product. At the same time, amine 31(5.0002g, 10.78mmol) was dissolved in DMF (14.15g, 15mL) and gently warmed to 30 ℃ to aid in material dissolution. The solution was allowed to cool to 20-25 ℃. The activated pyrazine solution was also cooled to about 15 ℃. A solution of compound 31 is added to the activated pyrazinecarboxylic acid while maintaining the temperature at 30 ℃ for about 1 hour. The solution was allowed to cool to 20-25 ℃ and then potassium carbonate (0.25g) in water (100mL) was added at 0 ℃. The mixture was filtered and washed with water (4 times 50mL each). The filter cake was dried under vacuum, starting at 20-25 ℃ and warmed to 30 ℃ after 24 hours until the filter cake was constant weight to give the title compound.

¹H NMR(DMSO-d₆，500MHz)：δ9.19ppm(1H，d，J＝1.3Hz)，8.90ppm(1H，d，J＝2.5Hz)，8.76ppm(1H，dd，J＝2.4Hz，1.5Hz)，8.50ppm(1H，d，J＝9.2Hz)，8.22ppm(1H，d，J＝9.0Hz)，4.68ppm(1H，dd，J＝9.1Hz，6.6Hz)，4.53ppm(1H，d，J＝9.0Hz)，3.96ppm(1H，d，J＝4.2Hz)，3.73ppm(1H，dd，J＝10.5Hz，7.5Hz)，3.68ppm(1H，dd，J＝10.6ppm，3.4ppm)，2.68-2.74ppm(1H，m)，2.52-2.58ppm(1H，m)，1.70-1.88ppm(3H，m)，1.51-1.69ppm(7H，m)，1.31-1.44ppm(2H，m)，1.39ppm(9H，s)，1.00-1.19ppm(4H，m)，0.97ppm(9H，s)，0.91-0.97ppm(1H，m).

Method 2

Oxalyl chloride (11.29mL) was added to a solution of pyrazine-2-carboxylic acid 32 and N-methylmorpholine (59.28mL) in dichloromethane (150mL) at about 30 ℃. The mixture was stirred for 0.5 h, then a solution of amine 31(50.0g) in dichloromethane (150mL) was added at about 30 ℃. After 0.5 h, the mixture was washed with water (250 mL). The aqueous phase was extracted with dichloromethane (100mL) to give a solution of the title compound in dichloromethane which was used directly in the next step (example 11, method 2).

Example 11: (1S) -2- ((S) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid (34)

Method 1

Concentrated HCl (150g, 0.015mol, 1.2 molar equivalents) was slowly added to a stirred solution of pyrazinyl peptide 33(50.0g) in formic acid (100.0g) at 0 ℃. After 3.3 hours, the reaction mixture was diluted with 166.5g of ice water. Dichloromethane (100mL) was added and the reaction stirred for 10 minutes to dissolve the product. The phases were separated and the aqueous layer was extracted with dichloromethane (100 mL). The combined organic phases were washed with water (75mL) and then concentrated to about 1/3 volumes at 50 ℃, 1 atm. Toluene (100mL) was added at room temperature and the homogeneous solution was evaporated under vacuum at 56 ℃ or less to about 1/3 volumes. The mixture was cooled to 20-25 ℃ and a precipitate formed. Heptane (75mL) was added slowly and the slurry was stirred for 10-15 minutes. The slurry was filtered and the filter cake was washed with heptane (50 mL). The solid was dried under vacuum at 20-25 ℃ to give compound 34.¹H NMR(500MHz，DMSO-d6)0.88-1.20(m，5H)，0.99(s，9H)，1.31-1.91(m，12H)，2.52(m，1H)，2.61(m，1H)，3.70(m，2H)，4.09(d，J＝4.2Hz，1H)，4.57(d，J＝9.0Hz，1H)，4.72(dd，J＝9.1，6.6Hz，1H)，8.28(d，J＝9.0Hz，1H)，8.52(d，J＝9.2Hz，1H)，8.77(dd，J＝2.4，1.5Hz，1H)，8.91(d，J＝2.5Hz，1H)，9.22(d，J＝1.3Hz，1H)，12.55(br s，1H).

Method 2

The dichloromethane solution of the starting compound 33 of example 10, method 2, was cooled to 0-5 ℃ and concentrated HCl (200mL) was then added while maintaining the temperature < 10 ℃. The mixture was stirred for 3 hours, then diluted with water (200mL) while maintaining a temperature < 10 ℃. The phases were separated and the aqueous phase was extracted with dichloromethane (100 mL). The combined organic phases were washed with water (100mL) and the aqueous phase was extracted with dichloromethane. The combined organic extracts were refluxed in a reverse Dean-Stark trap to azeotropize water. The mixture was concentrated to a minimum volume by distillation, then diluted with toluene (500mL) and then concentrated to 250mL by distillation at atmospheric pressure. The mixture was slowly cooled to 20 ℃ over about 6 hours. The resulting slurry was filtered and the filter cake was washed with toluene (100mL) and then dried in a vacuum oven at about 45 ℃ to provide the title compound as a light yellow powder containing about 17% toluene.

Example 12: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) -N- ((3S) -1- (cyclopropylamino) -2-hydroxy-1-oxohex-3-yl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide (35)

Method 1

A500 mL 3-necked round bottom flask equipped with an overhead stirrer, condenser, thermocouple, and nitrogen outlet was purged with nitrogen for a few minutes. Peptide-acid 34(25.0g, 0.049mol), EDC-HCl (10.35g, 0.054mol, 1.1 molar equivalent) and HOBt-H₂O (8.27g, 0.054mol, 1.1 mol equivalent) was added to the flask followed by 175mL of dichloromethane. The mixture was stirred at room temperature for 1 hour, then a suspension of hydroxyamide-amine 18(11.1g, 0.054mol, 1.1 molar equivalents) in dichloromethane (75mL) was added over 20 minutes while maintaining the temperature below 10 ℃. After the addition was complete, N-methylmorpholine (5.94mL, 0.054mol, 1.1 molar equivalent) was added in 2 portions. The mixture was allowed to warm to room temperature and stirred for 3 hours. NaHCO added to 200mL of water₃(8.0g) the reaction was quenched. The phases were separated and the organic layer was washed with water (175mL), 0.5N aq hcl (200mL), water (3 times 200mL each) and saturated NaCl (200mL) to give a 16% by weight solution of the title compound 35 in dichloromethane.

Method 2

N-methylmorpholine (38.19mL, 347.3mmol) was added to a mixture of peptide-acid 34(100.0g, 89.2 wt%, 173.7mmol), HOBt hydrate (26.79g, 87.6 wt%, 173.7mmol), EDCI (36.62g, 191.04mmol) and hydroxyamide-amine 18 in dichloromethane over 30 minutes while maintaining a temperature of 0-5 ℃. After the addition, the mixture was warmed to 20 ℃ and stirred for 5 hours. The mixture was then diluted with water (500mL) and stirred for about 0.5 hours. The phases were separated and the organic phase was washed with 1N HCl (500mL), 5 wt% aqueous sodium bicarbonate (500mL) to give a solution of the title compound in dichloromethane with 98.5% AUC purity, 95% solution yield.

Method 3

Peptidic acid 34(1.00 equiv.), EDCI (1.10 equiv.), HOBt hydrate (1.00 equiv.), and hydroxylamine 18. HCl (1.05 equiv.) were suspended in CH₂Cl₂(5 volumes) the mixture was cooled to 0-5 ℃. NMM (2.0 equivalents) was added over 30-60 minutes while maintaining the reaction temperature below 5 ℃. The reaction mixture was warmed to 20-25 ℃ over 30 minutes and stirred for 5 hours. The reaction was washed with water (5 vol), 1N HCl (5 vol) and 5 wt% aqueous NaHCO₃(5 vol) wash to provide the title compound in CH₂Cl₂The solution of (1).

Example 13: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) -N- ((S) -1- (cyclopropylamino) -1, 2-dioxohex-3-yl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide (4)

Method 1

A500 mL 3-necked round bottom flask equipped with an overhead stirrer, condenser, thermocouple, and nitrogen outlet was purged with nitrogen several times. Hydroxy amide peptide amide 35(128.64g, 16-17 wt%, 20.6g and 30mmol of 35) was dissolved in dichloromethaneThe solution was added to a reaction flask followed by 15% w/w aq. NaBr (13mL) and 7.5% w/w aq. NaHCO₃(52 mL). The solution was cooled to 5 ± 3 ℃ in an ice bath. TEMPO (0.7g) dissolved in dichloromethane (3mL) was added to the reaction mixture. In a separate Erlenmeyer flask, a 10-13% NaOCl solution (23.25mL, titer 108mg/mL, 2.51g, 33.7mmol, 1.12 molar equivalents) was diluted with water (70 mL). The NaOCl solution was added to the reaction mixture via an addition funnel at a rate to maintain the temperature below 8 ℃. The reaction mixture was allowed to stir at 5. + -. 3 ℃ for 1 hour. The layers were separated and the organic layer was washed with 10% (w/w) aq₂SO₃Quench (100mL) and wash with water (100 mL). The organic phase was reduced to dryness under reduced pressure and the solid was triturated with ethyl acetate (100mL) and filtered on a buchner funnel to give the title compound.

Method 2

TEMPO (1.09g, 6.95mmol) is added to a solution of 35 in dichloromethane of example 12, method 2, followed by a solution of sodium bicarbonate (21.89g, 260.5mmol) in water (400mL) and the mixture is cooled to 0-5 ℃. A solution of sodium hypochlorite (122.17g, 11.64 wt%, 191.04mmol) was added over 2 hours while maintaining the temperature at 0-5 ℃. The mixture was stirred at 0-5 ℃ for 1 hour, then the phases were separated. The organic phase was washed with water (500mL), 1 wt% aqueous sodium bisulfite (500mL), and water (500mL), then polish filtered (polish filter). The mixture was distilled at 38-42 ℃ with 710mm Hg to a volume of about 320 mL. Ethyl acetate (44mL) was added followed immediately by 1.5g of 4 seed crystals and the mixture was stirred at 38-42 ℃ for 15 minutes. Ethyl acetate (800mL) was added over 3 hours while maintaining the temperature at 38-42 ℃. The mixture was then distilled at 38-42 ℃ and 200-. Additional ethyl acetate (200mL) was added over 0.5 hour. The resulting slurry was cooled to 20-25 ℃ over 1 hour and stirred at the same temperature for an additional 1 hour. The mixture was filtered and the filter cake was washed with ethyl acetate (2 times 300mL each) and dried under vacuum at 45-55 ℃ with a nitrogen bleed (bleed) to give the title compound 4 as a white solid.

Method 3

TEMPO (0.06 eq) of 35 CH of example 12, method 3₂Cl₂Within the solution, the solution was stirred at 20-25 ℃ until all TEMPO had dissolved. To this solution NaHCO was added₃(1.5 equiv.) solution in water (4 vol.). The resulting biphasic mixture was cooled to 0-5 ℃. While maintaining the reaction temperature at 0-5 ℃, 10-13 wt% NaOCl solution (1.10 eq) was added over 2-3 hours and the mixture was stirred for an additional 1 hour. Separating the layers, and subjecting the organic layer to H at 0-5 deg.C₂O (5 vol.), 1 wt.% Na₂SO₃(5 vol.) and H₂O (5 vol) wash. Glacial acetic acid (0.12 eq) was added to compound 4 in CH₂Cl₂To stabilize compound 4 in solution.

Example 14: recrystallization of the compound of formula 4.

The solution of compound 4 from example 13, method 3 was filtered through Celite and the filtrate solution was reduced to 3.1-3.3 volumes by vacuum distillation below 20 ℃. After distillation, the solution was brought to 38-42 ℃ and then EtOAc (0.80 vol) was added followed by seeding with compound 4 (1.5 wt%, relative to 34 of example 12). The resulting mixture was stirred at 38-42 ℃ for 15 minutes. EtOAc (8 vol) was added to the mixture over 3 hours while maintaining the temperature at 38-42 ℃. The total volume of the slurry was then reduced to 3.9-4.1 volumes by vacuum distillation at 38-42 ℃. To the mixture was added EtOAc (2 vol) over 30 min while maintaining the batch temperature at 38-42 ℃. The resulting slurry was then cooled to 20-25 ℃ over 1 hour and stirred at 20-25 ℃ for an additional 1 hour. The slurry was filtered. The filter cake was washed with EtOAc (2X 3 volumes each) and dried under vacuum at 45-55 deg.C for 6 hours with a nitrogen purge.

Adding 2.2-2.4 volumes of CH to the dried filter cake₂Cl₂To a total volume of 3.1-3.3 volumes. The mixture was brought to 38-42 ℃ to obtain a homogeneous solution. EtOAc (0.80 vol) was added followed by seeding with compound 4 (1.5 wt%, relative to 34 for example 12). The resulting mixture was stirred at 38-42 ℃ for 15 minutes. EtOAc (8 vol) was added to the mixture over 3 hours while maintaining the temperature at 38-42 ℃. Then reducing the total volume of the slurry by vacuum distillation at 38-42 deg.CDown to 3.9-4.1 volumes. To the mixture was added EtOAc (2 vol) over 30 min while maintaining the batch temperature at 38-42 ℃. The resulting slurry was then cooled to 20-25 ℃ over 1 hour and stirred at 20-25 ℃ for an additional 1 hour. The slurry was filtered. The filter cake was washed with EtOAc (2 times 3 volumes each) and dried under vacuum at 45-55 ℃ for 12 hours with a nitrogen purge to give purified compound 4.¹H NMR (500MHz, CDCl3)0.78(m, 2H), 0.87(m, 2H), 0.91(s, 9H), 0.91(t [ shaded ]]，3H)，0.98(m，4H)，1.08(m，1H)，1.20(m，4H)，1.29(m，1H)，1.40(m，1H)，1..42(m，2H)，1.46(m，1H)，1.48(m，1H)，1.60(m，1H)，1.70(m，1H)，1.79(m，1H)，1.83(m，2H)，1.88(m，1H)，1.94(m，1H)，2.67(m，1H)，2.89(bs，1H)，2.96(bs，1H)，3.63(d，1H)，3.99(d，1H)，4.70(s，1H)，4.82(d，1H)，4.89(t，1H)，5.65(bs，1H)，7.74(bs，1H)，8.00(bs，1H)，8.06(bs，1H)，8.29(bs，1H)，8.60(s，1H)，8.77(s，1H)，9.42(s，1H).

Example 15: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) -N- ((S) -1- (cyclopropylamino) -1, 2-dioxo-hex-3-yl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide (4)

Step a: trans-N-cyclopropyl-2-hexanamide (37)

A flask equipped with an overhead stirrer, thermometer and addition funnel was placed under a nitrogen atmosphere, then (E) -hex-2-enoic acid (89.8g, 787mmol), EDCI (158.3g, 826mmol), HOBt (112.0g, 826mmol) and IPAc (890mL) were added, followed by cooling to 0. + -. 5 ℃. NMM (99.1mL, 1.6mol) was added to the addition funnel, which was then addedThe reaction mixture was added while maintaining the temperature at 0. + -. 5 ℃. The mixture was stirred for 30 min, then cyclopropylamine (60.0mL, 866mmol) was added and the reaction allowed to warm to 25. + -. 5 ℃ over 16 h. The reaction mixture was washed by adding hydrochloric acid (500mL, 1.0N), the mixture was vigorously stirred for 30 minutes, and then allowed to stand for 30 minutes; the layers were separated and the washing procedure was repeated. Sodium hydroxide (500mL, 1.0N) was added, then the mixture was stirred vigorously for 30 minutes, and then allowed to stand for 30 minutes; the layers were separated and the caustic wash procedure was repeated. Water (500mL) was added, and then the mixture was stirred vigorously for 30 minutes, and then allowed to stand for 30 minutes; the layers were separated and the washing procedure was repeated. The combined organic phases were concentrated under reduced pressure to 1/3 original volume, followed by the addition of IPAc (600 mL); this was repeated 2 times, at which time a white precipitate formed. The slurry was then concentrated to 2/3 original volume at atmospheric pressure and then cooled to 50 ± 5 ℃. N-heptane (890mL) was added slowly while the reaction was cooled to-5. + -. 5 ℃ and held at this temperature for 4 hours. The solid was filtered, washed with cold n-heptane (2 × 250mL) and dried to afford compound 37 as a fine white solid.¹H NMR(500MHz，d₆-DMSO)7.89(s，1H)，6.58(dt，J＝15.2，7.0Hz，1H)，5.80(dt，J＝15.2，1.3Hz，1H)，2.70-2.65(m，1H)，2.12-2.06(m，2H)，1.44-1.37(m，2H)，0.88(t，J＝7.3Hz，3H)，0.64-0.60(m，2H)，0.42-0.38(m，2H).

Step b: n-cyclopropyl-3-propyloxirane-2-carboxamide (38) or (2S, 3R) -N-cyclopropyl-3-propyloxirane-2-carboxamide (39)

Method 1

A flask equipped with an overhead stirrer, thermometer and addition funnel was placed under a nitrogen atmosphere, followed by addition of t-butyl hydroperoxide (TBHP; 95mL, 5.5M, 522mmol) and tetrahydrofuran (THF; 200 mL). The reaction was cooled to-20. + -. 5 ℃ and n-butyllithium (n-BuLi; 235mL, 2.5M, 587mmol) was added to the addition funnelSlowly adding the mixture, and keeping the reaction temperature to be lower than-5 +/-5 ℃. After the addition was complete, the reaction was warmed to 0. + -. 5 ℃ and Compound 37(19.80g, 130mmol) in THF (20mL) was added, the temperature was maintained at 0. + -. 5 ℃ and then the temperature was raised to 25. + -. 5 ℃ and the reaction stirred for 12 h. After this time IPAc (200mL) and saturated aqueous sodium bisulfite (200mL) were added and the reaction stirred for 60 min. The layers were separated and the aqueous layer was extracted with IPAc (2X 75mL each). The combined organic phases were washed with sodium sulfate (Na)₂SO₄) Dried, filtered and concentrated under reduced pressure to provide the title compound.¹H NMR(500MHz，DMSO-d6)：0.43(m，2H)，0.59(m，2H)，0.91(t，J＝7Hz，3H)，1.32-1.59(m，4H)，2.62(m，1H)，2.96(m，1H)，3.10(d，J＝2Hz，1H)，7.99(br s，1H).

Method 2 (stereospecificity)

A flask equipped with a stirring rod, a thermometer and an addition funnel was placed under a nitrogen atmosphere, and then samarium (III) isopropoxide (Sm (O-i-Pr) was added₃430mg, 1.3mmol), triphenylarsine oxide (Ph)₃As ═ O; 420mg, 1.3mmol), S- (-)1, 1' -bis-2-naphthol ((S) -BINOL), 370mg, 1.3mmol),Molecular sieves (13g) and THF (20mL) were then stirred at 25. + -. 5 ℃ for 30 minutes. T-butyl hydroperoxide (2.8mL, 5.5M, 16mmol) was then added. The mixture was stirred at 25. + -. 5 ℃ for 30 minutes, then compound 37(2.0g, 13mmol) in THF (2.0mL) was added. The reaction was stirred for 14 hours, after which time the reaction had reached 95% completion as determined by HPLC. The reaction mixture was diluted with ethyl acetate (100mL) and quenched by the addition of 10% citric acid solution (50 mL). The organic phase was separated and filtered through Celite. Distillation of the solvent under reduced pressure gave compound 39 as a pale yellow oil. Of the product¹H NMR is essentially the same as for racemic compound 38.

Method 3

To a reactor equipped with a mechanical stirrer and containing CH at 0 DEG C₂Cl₂A3-necked 250mL round bottom flask of compound 37(10.0g, 65.3mmol) and Urea Hydrogen Peroxide (UHP) (25.0g, 4.0 equivalents) in (100mL, 10 volumes) was charged with trifluoroacetic anhydride (41.1g, 27.2mL, 3.0 equivalents). The reaction mixture was heated to 35. + -. 5 ℃ and stirred for 2 hours. After allowing the reaction mixture to cool to room temperature, another aliquot of trifluoroacetic anhydride (13.7g, 9.0mL, 1.0 equiv.) was added. The reaction mixture was heated to 35. + -. 5 ℃ and stirred for a further 3 hours. The reaction mixture was then cooled again to 0 ℃ by slow addition of saturated NaHCO₃(5 vol.) quench and stir for 30 min. Separating the organic layer with CH₂Cl₂The aqueous layer was extracted (50mL, 5 volumes). The combined organic layers were dried and evaporated to give the crude product, N-cyclopropyl-3-propyloxirane-2-carboxamide (38), as a pale yellow oil. The crude product was used in the next step without further purification.

Step c: 3-azido-N-cyclopropyl-2-hydroxyhexanamide (40)

A flask equipped with an overhead stirrer, thermometer and reflux condenser was placed under a nitrogen atmosphere, and then compound 38(20.0g, 118mmol), sodium azide (NaN) and sodium azide were added₃(ii) a 31.0g, 473mmol), magnesium sulfate (MgSO₄(ii) a 14.0g, 118mmol) and methanol (MeOH; 200 mL). The mixture was heated to 65. + -. 5 ℃ for 2 hours and then cooled to 25. + -. 5 ℃ and filtered through a pad of Celite 545. The solvent was removed under reduced pressure to yield a thick oil, which was taken up in IPAc (250mL) and then washed with water (3X 250 mL). The organic phase was washed with sodium sulfate (Na)₂SO₄) Dried, filtered and concentrated under reduced pressure to afford compound 40 as a white solid.¹H NMR(500MHz，d₆-DMSO)7.87(s，1H)，5.97(d，J＝6.0，1H)，4.02(dt，J＝6.0，3.8Hz，1H)，2.70-2.65(m，1H)，1.60-1.20(m，4H)，0.88(t，J＝7.0Hz，3H)，0.63-0.58(m，2H)，0.51-0.46(m，2H).

Step d: 3-amino-N-cyclopropyl-2-hydroxyhexanamide (41)

Compound 40(15.1g, 71.3mmol), Pd/C (1.5g, 5 wt%, 50% wet), and MeOH (150mL) were added to the pressure vessel, which was then purged with nitrogen for 5 minutes. The vessel was sealed, pressurized to 1bar with nitrogen and then released 3 times. The same procedure was repeated with hydrogen. After the 3 rd purge with hydrogen, 3bar of 3 hydrogen was added to the vessel at a pressure of 3 bar. The reaction was stirred and maintained at a temperature of 25 ± 5 ℃ for 14 hours, after which the reaction mixture was filtered through a pad of Celite 545 and the solvent was removed to afford crude compound 41(8.48g) as a yellow solid. Acetonitrile (ACN; 150mL) was added to the material and the reaction was heated to reflux, at which time all solids dissolved. The mixture was then cooled to 25 ± 5 ℃, the white needles formed were collected, washed with cold ACN and dried to afford purified compound 41.¹H NMR(500MHz，d₆-DMSO)：8.05(br s，3H)，4.20(d，J＝3.2，1H)，3.42-3.34(m，1H)，2.71-2.65(m，1H)，1.51-1.20(m，4H)，1.17(d，J＝6.5Hz，1H)，0.83(t，J＝7.6Hz，3H)，0.64-0.60(m，2H)，0.54-0.49(m，2H).

Step e: 3-amino-N-cyclopropyl-2-hydroxyhexanamide deoxycholate (42).

To a 3-neck 250mL round bottom flask equipped with a mechanical stirrer and containing racemic 3-amino-N-cyclopropyl-2-hydroxyhexanamide 41(10.0g, 53.69mmol) in THF (100mL) was added deoxycholic acid (15.8g, 40.27mmol, 0.75 equiv). The reaction mixture was stirred at 65. + -. 5 ℃ for 2 hours. The resulting homogeneous mixture was allowed to cool to a temperature of 22-25 ℃ over 1 hour and then maintained at that temperature range for an additional 4 hours. The precipitated solid was collected by filtration, washed with THF (10mL), and dried overnight to give 12.2g of 3-amino-N-cyclopropyl-2-hydroxyhexanamide deoxycholate 42 as a white solid.

Step f: 3-amino-N-cyclopropyl-2-hydroxyhexanamide hydrochloride (43).

To a 3-neck 250mL round bottom flask equipped with a mechanical stirrer and containing a mixture of deoxycholate (42) in 2-propanol (62mL) was added a 5-6N HCl solution in isopropanol (66mL, 3 equiv) with stirring. The resulting solution was heated at 75 + -5 deg.C for 1 hour, allowed to cool to a temperature of 22-25 deg.C over 1 hour, and then maintained at that temperature range for an additional 4 hours. The precipitated solid was collected by filtration, washed with 2-propanol (12mL, 1 volume) and dried overnight to give 7.2g of hydrochloride salt 43 as a white solid.

Step g: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) -N- ((2S, 3S) -1- (cyclopropylamino) -2-hydroxy-1-oxohex-3-yl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide (36)

Will CH₂Cl₂(6 volumes) are added to a vessel and cooled to 0-5 ℃. Carboxylic acid 34(1.00 eq.), HOBt activator (1.10 eq.), EDCI (1.10 eq.), andamine-hydrochloride 43(1.10 equivalents) was added to the same vessel in this particular order. NMM (2.0 equivalents) was added over 30-60 minutes while maintaining the reaction temperature below 5 ℃. The reaction mixture was warmed to 20-25 ℃ over 30 minutes and stirred for an additional 12 hours. The reaction was washed with water (5 vol), 1N HCl (5 vol) and 5 wt% aqueous NaHCO₃(5 volumes) washing to give chiral hydroxy-peptide 36 in CH₂Cl₂The solution of (1).¹H NMR(500MHz，DMSO-d6).0.47(m，2H)，0.58(m，2H)，0.78(t，J＝7.2Hz，3H)，0.80-1.83(m，21H)，0.94(s，9H)，2.58(m，1H)，2.65(m，1H)，2.70(m，1H)，3.64(dd，J＝3.2，10.6Hz，1H)，3.77(dd，J＝7.8.10.2Hz，1H)，3.80(dd，J＝4.0，5.8Hz 1H)，4.04(m，1H)，4.26(d，J＝3.0Hz，1H)，4.56(d，J＝9.2Hz，1H)，4.69(dd，J＝6.4，9.0Hz，1H)，5.56(d，J＝5.8Hz，1H)，7.58(d，J＝9.1Hz，1H)，7.69(d，J＝4.6Hz，1H)，8.23(d，J＝9.2Hz，1H)，8.51(d，J＝9.1Hz，1H)，8.77(dd，J＝1.5，2.4Hz，1H)，8.91(d，J＝2.5Hz，1H)，9.19(d，J＝1.4Hz，1H).

Step h: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) -N- ((S) -1- (cyclopropylamino) -1, 2-dioxohex-3-yl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide (4)

The final CH containing chiral hydroxy-peptide 36 of step g of this example was reacted at 20-25 deg.C₂Cl₂Adding NaHCO into the solution₃(1.5 equiv.) in water (4 vol). TEMPO (0.06 eq.) was added to the mixture. The resulting biphasic mixture was cooled to 0-5 ℃. While maintaining the reaction temperature at 0-5 deg.C, 10-18 wt% NaOCl solution (1.10 equivalents) was added over 2-3 hours and stirred for an additional 1 hour. The layers were separated and the organic layer was washed with aqueous 5 wt% NaCl solution (5 vol.) and aqueous 1 wt% Na at 0-5 deg.C₂SO₃And aqueous 5% by weightNaCl solution (5 volumes) wash. Glacial acetic acid (0.12 eq) was added and the resulting compound 4 was in CH₂Cl₂The solution in (1) continues to the next step.

Step i: double recrystallization of (1S, 3aR, 6aS) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) -N- ((S) -1- (cyclopropylamino) -1, 2-dioxohex-3-yl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide (4).

The organic layer of step h of this example was reduced to 3.1-3.3 volumes by vacuum distillation at a temperature less than or equal to 20 ℃. After distillation, the solution was brought to 38-42 ℃. EtOAc (0.80 vol) was added followed by seed crystals of compound 4 (1.5 wt% relative to carboxylic acid 34 added in step g of this example). The resulting mixture was stirred at 38-42 ℃ for 15 minutes. EtOAc (8 vol) was added to the mixture over 3 hours while maintaining the temperature at 38-42 ℃. The total volume of the slurry was then reduced to 3.9-4.1 volumes by vacuum distillation at 30-40 ℃. EtOAc (2 vol) was added to the mixture over 30 min while maintaining the batch temperature at 38-42 ℃. The resulting slurry was then cooled to 20-25 ℃ over 1 hour and stirred at 20-25 ℃ for an additional 1 hour. The slurry was filtered. The filter cake was washed with EtOAc (2X 3 vol) and dried under vacuum at 45-55 ℃ for 5 h with a nitrogen purge.

The filter cake was cooled to ambient temperature, transferred to a flask where it was dissolved in CH₂Cl₂(3.7 vol.). Compound 4 is in CH₂Cl₂The solution in (a) was filtered and then transferred into the reactor. Using CH for flask₂Cl₂(1.3 vol) wash, filter wash and transfer to batch. The organic layer is reduced to 3.6-3.8 volumes by vacuum distillation at 20 ℃ or less. After distillation, the solution was brought to 38-42 ℃. EtOAc (0.93 vol) was added followed by seed crystals of compound 4 (1.73 wt% relative to carboxylic acid 34 added in step g of this example). The resulting mixture was stirred at 38-42 ℃ for 15 minutes. EtOAc (9.4 vol) was added to the mixture over 3 hours while maintaining the temperature at 38-42 ℃. The total volume of the slurry was then reduced to 4 by vacuum distillation at 30-40 ℃.5-4.7 volume. EtOAc (2.3 vol) was added to the mixture over 30 min while maintaining the batch temperature at 38-42 ℃. The resulting slurry was then cooled to 20-25 ℃ over 1 hour and stirred at 20-25 ℃ for an additional 1 hour. The slurry was filtered. The filter cake was washed with EtOAc (2X 3.5 vol) and dried under vacuum at 45-55 ℃ for 8 h with a nitrogen purge. NMR data: see example 14.

Example 16: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) -N- ((S) -1- (cyclopropylamino) -1, 2-dioxo-hex-3-yl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide (4)

Step a: preparation of (2R, 3S) -3-amino-N-cyclopropyl-2-hydroxyhexanamide (44).

Racemic compound 24 (using the compound shown in scheme IV (paragraph [0151 ]]-[0152]) By HPLC) to its (2S, 3S) and (2R, 3S) diastereomers. The (2R, 3S) diastereomer of compound 24 (35.5g) was dissolved in methanol (355mL) and palladium hydroxide on carbon (Pearlman' S catalyst, 2.13g) was added. The system was flushed with nitrogen and then hydrogen was bubbled through for 2 hours. The solution was filtered and the solvent was then distilled. Ethyl acetate (250mL) was added 2 times and the mixture was again distilled to minimum volume. Ethyl acetate was again added and the solution was concentrated to about 100 mL. Heptane (100mL) was added and the resulting slurry was stirred at 0 ℃ for 30 minutes. Compound 44 was collected by filtration, washed with cold heptane and dried overnight at 40 ℃.¹H NMR[(2R，3S)24](500MHz，DMSO-d6)：0.43(m，2H)，0.57(m，2H)，0.86(t，J＝7.2Hz，3H)，1.12-1.55(m，4H)，3.76(m，1H)，3.82(m，1H)，5.00(m，2H)，5.35(d，J＝6.2Hz，1H)，6.58(br d，J＝6.2Hz，1H)，7.23-7.40(m，5H)，7.74(br d，J＝4.8Hz，1H).¹H NMR[44](500MHz，DMSO-d6)：0.47(m，2H)，0.59(m，2H)，0.86(t，J＝7.1Hz，3H)，1.10-1.45(m，4H)，2.65(m，1H)，2.79(m，1H)，3.63(d，J＝3.4Hz，1H)，7.72(br d，J＝3.8Hz，1H).

Step b: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) -N- ((2R, 3S) -1- (cyclopropylamino) -2-hydroxy-1-oxohex-3-yl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide (45)

Compound 44(4.0g, 7.8mmol) was dissolved in dichloromethane (35mL), the solution cooled to 10 deg.C and 1-hydroxybenzotriazole hydrate (1.36g, 8.9mmol) added. After stirring for 30 minutes, the temperature was further lowered to 0 ℃. N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (1.72g, 9.0mmol) was added and stirring was continued for 1 h. Compound 34(1.77g, 9.5mmol) was added and stirring was continued at 0 ℃ for 30 min. The mixture was allowed to warm to room temperature and stirred overnight. The mixture was then washed with aqueous sodium bicarbonate, dilute hydrochloric acid and brine in that order. The organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was dissolved in ethyl acetate, concentrated 3 more times, then dissolved in-4 volumes and cooled to 0 ℃. Heptane was added dropwise until the solid began to thicken, then the rate of addition was increased until a total of 50mL heptane was added. After stirring at 0 ℃ for 30 minutes, the product was collected by filtration. After drying overnight at 40 ℃, compound 45 was isolated as a white solid.¹H NMR(500MHz，DMSO-d6)0.43(m，2H)，0.57(m，2H)，0.82(t，J＝7.2Hz，3H)，0.8-1.8(m，21H)，0.93(s，9H)，2.58(m，1H)，2.65(m，1H)，2.63(dd，J＝3.2Hz，10.6，1H)，3.73(dd，J＝7.7，10.4Hz，1H)，3.81(dd，J＝2.7，5.9Hz，1H)，3.98(m，1H)，4.20(d，J＝3.0Hz，1H)，4.54(d，J＝9.3Hz，1H)，4.68(dd，J＝6.4，9.1Hz，1H)，5.50(d，J＝5.8Hz，1H)，7.32(d，J＝9.2Hz，1H)，7.59(d，J＝4.3Hz，1H)，8.21(d，J＝9.2Hz，1H)，8.50(d，J＝9.2Hz，1H)，8.76(dd J＝1.6，2.3Hz，1H)，8.90(d，J＝2.4Hz，1H)，9.19(d，J＝1.5Hz，1H).

Step c: (1S, 3aR, 6aS) -2- ((S) -2- ((S) -2-cyclohexyl-2- (pyrazine-2-carboxamido) acetylamino) -3, 3-dimethylbutyryl) -N- ((S) -1- (cyclopropylamino) -1, 2-dioxohex-3-yl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide (4)

Compound 45(2.0g, 2.9mmol) was dissolved in dichloromethane (14mL), to which was added a solution of sodium bromide (144mg) in water (1.0mL), followed by a solution of sodium bicarbonate (300mg) in water (4.0 mL). The mixture was cooled to 0 ℃ and TEMPO (53mg, 0.34mmol) was added. A commercial bleaching solution (3.3mL) containing sodium hypochlorite, further diluted with water (9.6mL), was added dropwise over the course of 10 minutes. The mixture was stirred at 5 ℃ for 1.5 h, then the organic layer was separated, washed with 10% aqueous sodium sulfite (8mL) then brine. The resulting solution was dried over magnesium sulfate, filtered, and then concentrated to dryness. Compound 4 showed the same spectral properties as those reported in example 14 after crystallization from dichloromethane/ethyl acetate.

Claims

1. A process for preparing a racemic mixture of cis-and trans-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acids of formula 7

wherein R' is C_1-5An alkyl group.

2. The process of claim 1, wherein the carboxylation step comprises forming the 2-anion of the compound of formula 6 in the presence of a complexing agent,

and treating the 2-anion with carbon dioxide to produce a racemic mixture of formula 7 trans-/cis-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid

3. The process of claim 2, wherein the 2-anion is prepared by treating the compound of formula 6 with a strong lithium base in the presence of a complexing agent and an aprotic solvent.

4. The process of claim 3, wherein the base is sec-butyllithium.

5. The method of claim 4, wherein the complexing agent is tetramethylethylenediamine, tetraethylethylenediamine, tetramethyl-1, 2-cyclohexyldiamine, sparteine, or 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] nonane.

6. The process of claim 2 wherein the complexing agent is tetramethylethylenediamine, tetraethylethylenediamine, tetramethyl-1, 2-cyclohexyldiamine, or 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] nonane.

7. The process of claim 2, wherein the trans-/cis-ratio is 1: 1.

8. The process of claim 2, wherein the trans-/cis-ratio is 60: 40.

9. The process of claim 2, wherein the trans-/cis-ratio is 80: 20.

10. The process of claim 2, wherein the trans-/cis-ratio is 90: 10.

11. The process of claim 2 wherein the trans-/cis-ratio is greater than 98: 2.

12. The method of claim 2, wherein the complexing agent is D-sparteine.

13. The process of claim 3 wherein the lithium base is sec-butyllithium and the complexing agent is 3, 7-dipropyl-3, 7-diazabicyclo [3.3.1] nonane to provide a mixture of racemic trans-/cis-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid of formula 7 wherein the trans-/cis-ratio is greater than 90: 10.

14. The process of claim 13 wherein the trans-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid is trans-N-tert-butoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid.

15. A process for preparing a compound of formula 4

The method comprises the following steps:

i) providing N-alkoxycarbonyl-3-azabicyclo [3.3.0] octane;

v) forming a salt with an optically active amine;

vi) crystallizing the salt;

vii) esterifying the acid provided in step vi);

29

xvi) oxidizing the hydroxy group of formula 35 to produce the compound of formula 4

16. The process of claim 15, wherein the oxidizing agent used in step xvi) is sodium hypochlorite and the oxidation is carried out in the presence of 2, 2, 6, 6-tetramethylpiperidinyloxy radical (TEMPO).

17. The process of claim 15 wherein the oxidizing agent used in step xvi) is 1, 1-dihydro-1, 1, 1-triacetoxy-1, 2-benziodoxazol-3 (1H) -one.

18. The method of claim 15, further comprising dissolving the compound of formula 4 in an organic solvent to obtain a solution of the compound of formula 4, and then adding an acid to the solution.

19. The process of claim 18, wherein the organic solvent is dichloromethane and the acid is acetic acid.

20. The method of claim 18, further comprising concentrating the solution of the compound of formula 4 to obtain the compound in solid form.

21. A method of purifying a compound of formula 4, the method comprising:

ii) adding an acid to the solution of the compound of formula 4, and

22. The process of claim 21, wherein the organic solvent is dichloromethane and the acid is acetic acid.

23. A process for preparing a compound of formula 8

The process comprises carboxylating an azabicyclooctane of formula 6

Balancing the trans-/cis-mixture of the compound of formula 7 in the presence of a suitable base to produce a trans-dominant-cis racemic acid of formula 8, wherein the trans-/cis-ratio in the compound of formula 8 is greater than 80: 20, and R' is C_1-5An alkyl group.

24. The process of claim 23 further comprising balancing a trans-/cis-mixture of the compound of formula 7 in the presence of a suitable base to produce a trans-predominately cis racemic acid of formula 8 wherein the trans-/cis-ratio is greater than 90: 10.

25. The process of claim 23, further comprising balancing the trans-/cis-mixture of formula 7 in the presence of a suitable base to produce a trans-predominately cis racemic acid of formula 8, wherein the trans-/cis-ratio is greater than 98: 2.

26. The process of claim 25, wherein the base is lithium hexamethyldisilazide, lithium diisopropylamide, or lithium 2, 2, 6, 6-tetramethylpiperidine.

27. The process of claim 26, wherein the base is lithium hexamethyldisilazide.

28. A process for preparing (1S, 2S, 3R) trans-N-alkoxycarbonyl-octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid comprising carboxylating an azabicyclooctane of formula 6

29. The method of claim 28, wherein the separating comprises the steps of:

i) forming a salt with an optically active base; and

ii) crystallizing the salt formed in step i) to provide an optically active salt of formula 9

9

30. The process of claim 29 wherein the optically active base is (R) α -aminoethylbenzene.

31. The process of claim 29 wherein the optically active base is (S)1, 2, 3, 4-tetrahydro-1-naphthylamine.

32. A process for preparing a compound of formula 1, the process comprising carboxylating an azabicyclooctane of formula 6

With a compound containing R₃Compounds of the group esterify a carboxylic acid of formula 9; and removal of the-COOR' protecting group to produceCompounds of formula 1

33. The method of claim 32, wherein R₃Is a tert-butyl group.

34. A process for preparing a compound of formula 4

The method comprises the following steps:

ii) forming a salt with an optically active amine;

iii) crystallizing the salt;

iv) esterifying the acid provided in step iii);

vi) reacting the bicyclic amino ester of step v) with a protected amino acid of formula 26,

xiii) oxidizing the hydroxy group of formula 35 to produce the compound of formula 4

35. A process for preparing a compound of formula 4

The method comprises the following steps:

i) providing N-alkoxycarbonyl-3-azabicyclo [3.3.0] octane;

v) forming a salt with an optically active amine;

vi) crystallizing the salt;

vii) esterifying the acid provided in step vi);

xvi) oxidizing the hydroxy group of formula 36 to produce the compound of formula 4

36. A process for preparing a compound of formula 4

The method comprises the following steps:

i) providing N-alkoxycarbonyl-3-azabicyclo [3.3.0] octane;

v) forming a salt with an optically active amine;

vi) crystallizing the salt;

vii) esterifying the acid provided in step vi);

xvi) oxidizing the hydroxy group of formula 45 to produce the compound of formula 4

37. The process of claim 34, 35 or 36, wherein the oxidizing agent used in step xvi) is sodium hypochlorite and the oxidation is carried out in the presence of 2, 2, 6, 6-tetramethylpiperidinyloxy radical (TEMPO).

38. The process of claim 34, 35 or 36, wherein the oxidizing agent used in step xvi) is 1, 1-dihydro-1, 1, 1-triacetoxy-1, 2-benziodoxazol-3 (1H) -one.

39. The method of claim 34, 35 or 36, further comprising dissolving the compound of formula 4 in an organic solvent to obtain a solution of the compound of formula 4, and then adding an acid to the solution.

40. The process of claim 39, wherein the organic solvent is dichloromethane and the acid is acetic acid.

41. The method of claim 39, further comprising concentrating the solution of the compound of formula 4 to obtain the compound in solid form.

42. A compound of formula 4 prepared by the process of claim 15, 34, 35 or 36

43. A compound of formula 7 prepared by the process of claim 1

44. A compound of formula 8 prepared by the process of claim 23

45. A compound of formula 9 prepared by the process of claim 28

46. A compound of formula 1 prepared by the process of claim 32