CN101448781A - Deuterated hepatitis C protease inhibitors - Google Patents

Abstract

A deuterated a-ketoamido steric specific compound of the formula (I), wherein D denotes a deuterium atom on a steric specific carbon atom.

Description

The deuterated hepatitis C protease inhibitors

Cross reference

The application requires the U.S. Provisional Application sequence the 60/782nd of submission on March 16th, 2006, No. 778, the rights and interests that No. the 60/844th, 771, the U.S. Provisional Application sequence of submitting in No. the 60/782nd, 976, the U.S. Provisional Application sequence that on March 16th, 2006 submitted to and on September 15th, 2006.

Background of invention

It is a physianthropy problem of needing solution badly that hepatitis C virus (" HCV ") infects.HCV is acknowledged as the reason of most of non-A non-B hepatitis, and the serum prevalence rate of global population is people such as 3%[A.Alberti according to estimates, " Natural History of Hepatitis C; " J.Hepatology, 31., (Suppl.1), 17-24 page or leaf (1999)].Only in the U.S., nearly four million peoples may infected [MJ.Alter, " The Epidemiology of Viral Hepatitis in theUnited States, Gastroenterol.Clin.North Am., 23, the 437-455 pages or leaves (1994); M.J.Alter " Hepatitis C Virus Infection in the United States, " J.Hepatology, 31., (Suppl.1), 88-91 page or leaf (1999)].

As if be exposed to HCV when first, 20% the infected individual development of only having an appointment is acute clinical hepatitis, and other people infection is spontaneously dissipated.But, in 70% situation almost, virus is set up chronic infection, continues many decades [S.Iwarson, " The Natural Course ofChronic Hepatitis, " FEMS Microbiology Reviews, 14, the 201-204 pages or leaves (1994); D.Lavanchy, " Global Surveillance and Control of Hepatitis C, " J.ViralHepatitis, 6, the 35-47 pages or leaves (1999)].This causes recurrent and carrying out property deterioration liver inflammation usually, often cause even more serious morbid state, for example liver cirrhosis and hepatocellular carcinoma [M.C.Kew, " Hepatitis C and Hepatocellular Carcinoma ", FEMS MicrobiologyReviews, 14, the 211-220 pages or leaves (1994); People such as I.Saito, " Hepatitis C VirusInfection is Associated with the Development of HepatocellularCarcinoma, " Proc.Natl.Acad.Sci.USA, 87, the 6547-6549 pages or leaves (1990)].Unfortunately, there is not general effectively treatment can weaken the progress of chronic hcv.

HCV genome encoding 3010-3033 amino acid whose polyprotein [Q.L.Choo etc., ＂ Genetic Organization and Diversity of the Hepatitis C Virus. ＂ Proc.Natl.Acad.Sci.USA, 88, pp.2451-2455 (1991); N.Kato etc., ＂ MolecularCloning of the Human Hepatitis C Virus Genome From Japanese Patientswith Non-A, Non-B Hepatitis, ＂ Proc.Natl.Acad.Sci.USA, 87, pp.9524-9528 (1990); A.Takamizawa etc., ＂ Structure and Organization of theHepatitis C Virus Genome Isolated From Human Carriers, ＂ J.Virol., 65, pp.1 105-1 1 13 (1991)].HCV unstructuredness (NS) albumen is assumed that virus replication provides essential catalysis means.NS albumen originates from the proteolysis cracking [R.Bartenschlager etc. of polyprotein, ＂ Nonstructural Protein 3 of the Hepatitis C VirusEncodes a Serine-Type Proteinase Required for Cleavage at the NS3/4 andNS4/5 Junctions, ＂ J.Virol., 67, pp.3835-3844 (1993); A.Grakoui etc., ＂ Characterization of the Hepatitis C Virus-Encoded Serine Proteinase:Determination of Proteinase-Dependent Polyprotein Cleavage Sites, ＂ J.Virol., 67, pp.2832-2843 (1993); A.Grakoui etc., ＂ Expression andIdentification of Hepatitis C Virus Polyprotein Cleavage Products, ＂ J.Virol., 67, pp.1385-1395 (1993); L.Tomei etc., ＂ NS3 is a serine protease requiredfor processing of hepatitis C virus polyprotein ＂, J.Virol., 67, pp.4017-4026 (1993)].

HCV NS albumen 3 (NS3) contains serine protease, and this helps to process most of viral enzymes, thereby is regarded as virus replication and infectious necessary.The sudden change of known yellow fever virus NS3 proteolytic enzyme reduces the infectivity [Chambers of virus, people such as TJ, ＂ Evidence that theN-terminal Domain of Nonstructural Protein NS3 From Yellow Fever Virusis a Serine Protease Responsible for Site-Specific Cleavages in the ViralPolyprotein ＂, Proc.Natl.Acad.Sci.USA, 87, pp.8898-8902 (1990)].Preceding 181 amino acid (1027-1207 residue of viral polyprotein) that shown NS3 contain the NS3 serine protease structural domain [people such as C.Lin in the site, whole four downstreams of processing HCV polyprotein, ＂ Hepatitis C Virus NS3 Serine Proteinase:Trans-CleavageRequirements and Processing Kinetics ＂, J.Virol., 68, pp.8147-8157 (1994)].

HCV NS3 serine protease and relevant cofactor NS4A thereof help to process whole viral enzymes, thereby it is necessary to be regarded as virus replication.As if this processing is similar to the processing of being undertaken by human immunodeficiency virus's aspartyl protease, and this proteolytic enzyme also participates in viral enzyme processing.Hiv protease suppresses the processing of viral protein, and it is strong human antiviral agent, shows to disturb the viral life cycle in this stage can obtain the therapeutic activity agent.So HCV NS3 serine protease is an attractive drug discovery target.

At present also without any gratifying whose anti-HCV agent or treatment.Up to date, unique set HCV physics is an interferon therapy.But, Interferon, rabbit has pronounced side effects [M.A.Wlaker etc., ＂ Hepatitis C Virus:An Overview of Current Approaches andProgress, ＂ DDT, 4, pp.518-29 (1999); D.Moradpour etc., ＂ Current andEvolving Therapies for Hepatitis C, ＂ Eur.J.Gastroenterol.Hepatol., 11, pp.1199-1202 (1999); H.L.A.Janssen etc., ＂ Suicide Associated withAlfa-Interferon Therapy for Chronic Viral Hepatitis, ＂ J.Hepatol., 21, pp.241-243 (1994); P.F.Renault etc., ＂ Side Effects of Alpha Interferon, ＂ Seminars in Liver Disease, 9, pp.273-277. (1989)], only in a part (about 25%) case, induce secular mitigation [O.Weiland, ＂ Interferon Therapy in ChronicHepatitis C Virus Infection ＂, FEMS Microbiol.Rev., 14, pp.279-288 (1994)].Nearest PEGization form (PEG-INTRON about Interferon, rabbit

And PEGASYS

) and virazole and PEGization conjugation element (REBETROL

) the introduction of combination therapy, only aspect remission rate, produce a spot of improvement and aspect side effect, only produce part and reduce.And effectively the prospect of whose anti-HCV vaccine remains uncertain.

Thereby, there is demand to more effective whose anti-HCV therapy.This class inhibitor will have the treatment potentiality as proteinase inhibitor, especially as serpin, more specifically as HCV NS 3 proteinase inhibitor.Particularly, this compounds can be used as antiviral agent, in particular as the whose anti-HCV agent.

Deuterium is introduced in recent findings in compound will reduce epimerization speed owing to the isotopic effect of deuterium, thus with respect to its not the deuterated analogue improve the bulk concentration of active isomer.

Summary of the invention

The present invention relates to the compound of deuterated formula (I)

And pharmacologically acceptable salts, prodrug and solvated compounds.D represents D atom in (I).

When relating to formula (I),

D represents D atom;

R ¹Be

Wherein

Be optional monocycle azaheterocyclyl that replaces or the optional many rings azaheterocyclyl that replaces, or the optional many rings azepine heterocycloalkenyl that replaces, wherein unsaturated is away from having R ²¹The ring of part and with-C (O)-N (R ²)-CDR ³-C (O)-C (O)-NR ⁴R ⁵In the ring that part connects;

R ²¹Be Q ³-W ³-Q ²-W ²-Q ¹W wherein ²And W ³Be key independently of one another ,-CO-,-CS-,-C (O) N (Q ⁴)-,-CO ₂-,-O-,-N (Q ⁴)-C (O)-N (Q ⁴)-,-N (Q ⁴)-C (S)-N (Q ⁴)-,-OC (O) NQ ⁴-,-S-,-SO-,-SO ₂-,-N (Q ⁴)-,-N (Q ⁴) SO ₂-,-N (Q ⁴) SO ₂N (Q ⁴)-, and work as W ²And W ³When any one was end group, it was a hydrogen;

Q ¹, Q ²And Q ³Be key independently of one another, the optional aliphatic group that replaces, the optional assorted aliphatic group that replaces, the optional cyclic aliphatic base that replaces, the optional aryl that replaces, the optional heteroaryl that replaces, the optional aralkyl that replaces, or the optional heteroaralkyl that replaces; Or work as Q ³, Q ², or Q ¹When any one was end group, it was a hydrogen, and condition is to work as W ³And W ²Q when all existing ²It or not key; With

R ², R ³And R ⁴Be H or C independently of one another _1-6Alkyl; With

R ⁵Be H, alkyl, cycloalkyl, the optional aryl that is replaced by 1-4 alkyl, alkylaryl, aryl, the optional amino that is replaced by 1 or 2 alkyl; With

R ²¹Be Q ³-W ³-Q ²-W ²-Q ¹W wherein ²And W ³Be key independently of one another ,-CO-,-CS-,-C (O) N (Q ⁴)-,-CO ₂-,-O-,-N (Q ⁴)-C (O)-N (Q ⁴) ,-N (Q ⁴)-C (S)-N (Q ⁴)-,-OC (O) NQ ⁴-,-S-,-SO-,-SO ₂-,-N (Q ⁴)-,-N (Q ⁴) SO ₂-,-N (Q ⁴) SO ₂N (Q ⁴)-and work as W ²And W ³When any one was end group, it was a hydrogen; Q ¹, Q ²And Q ³Be key independently of one another, the optional aliphatic group that replaces, the optional assorted aliphatic group that replaces, the optional cyclic aliphatic base that replaces, the optional aryl that replaces, the optional heteroaryl that replaces, the optional aralkyl that replaces, the optional heteroaralkyl that replaces; Or work as Q ³, Q ², or Q ¹When any one was end group, it was a hydrogen, and condition is to work as W ³And W ²Q when all existing ²It or not key.

In some embodiments, R ¹Be

Wherein

R ⁶And R ⁸Be independently of one another

Key; Or

Optional replacement (1,1-or 1,2-) ring alkylidene group; Or

Optional replacement (1,1-or 1,2-) heterocycle alkylidene group (heterocyclylene); Or

Methylene radical or ethylidene, can be replaced by a substituting group that is selected from the group of forming by following group, described substituting group is the optional aliphatic group that replaces, optional cyclic group that replaces and the optional aromatic group that replaces, wherein methylene radical or ethylidene are further optional is replaced by the aliphatic group substituting group;

R ⁷, R ⁹And R ¹¹Be hydrogen or the optional aliphatic group that replaces independently of one another;

R ¹⁰Be the optional aliphatic group that replaces, optional cyclic group that replaces or the optional aromatic group that replaces;

L is-C (O)-,-OC (O)-,-NR ¹¹C (O)-,-S (O) ₂-,-NR ¹¹S (O) ₂-, or key;

N is O or 1.

In some embodiments, n is 1.

In some embodiments, R ⁶By a methylene radical that is selected from the substituting group replacement of the group of being made up of following group, described substituting group is the optional aliphatic group that replaces, the cyclic group of optional replacement, the optional aromatic group that replaces.

In some embodiments, R ⁶The methylene radical that is replaced by isobutyl-.

In some embodiments, R ⁷Be hydrogen.

In some embodiments, R ⁸By a methylene radical that is selected from the substituting group replacement of the group of being made up of following group, described substituting group is selected from the optional aliphatic group that replaces, and the optional cyclic group that replaces is chosen the aromatic group that replaces wantonly.In some other embodiments, R ⁸It is the methylene radical that the cyclic group that is optionally substituted replaces.Or in some other embodiments, R ⁸The methylene radical that is replaced by cyclohexyl.

In some embodiments, R ⁹Be hydrogen.

In some embodiments, L is-CO-.

In some embodiments, R ¹⁰It is optional substituted aromatic group.

In some embodiments, R ¹⁰Be selected from following group:

With

In some embodiments, R ¹⁰It is the optional pyrazinyl (for example 2-pyrazinyl) that replaces.

In some embodiments,

It is the monocycle azaheterocyclyl that replaces.

In some embodiments, Be the pyrrolidyl that the 3-carbon atom is replaced by heteroaryloxy, wherein heteroaryl is further optional is replaced by 1-4 halogen group.

In some embodiments,

Be

In some embodiments,

It is the optional many rings azaheterocyclyl that replaces.

In some embodiments,

Be

Or

In some embodiments,

Be

Or

In some embodiments, R ²Be hydrogen, R ⁴And R ⁵Be hydrogen or cyclopropyl independently of one another.In another embodiment, R ³It is propyl group.In another embodiment, n is 0.In another embodiment, L is-NR ¹¹C (O)-, R ¹¹Be hydrogen.In another embodiment, R ¹⁰It is the optional aliphatic group that replaces.In another embodiment, R ¹⁰It is the tertiary butyl.In another embodiment, this compound is

Or

In some embodiments, R ¹Be

Wherein A is-(CHX ¹) _a-;

B is-(CHX ²) _b-;

A is 0 to 3;

B is 0 to 3, and condition is that a+b is 2 or 3;

X ¹And X ²Independently be selected from hydrogen separately, the optional C that replaces _1-4Aliphatic group, the optional aryl that replaces;

Y ¹And Y ²Be hydrogen independently of one another, the optional aliphatic group that replaces, the optional aryl that replaces, amino or-OQ ⁴Q wherein ⁴Be hydrogen or the optional aliphatic group that replaces independently of one another;

R ²²Be the optional aliphatic group that replaces, the optional cyclic aliphatic base that replaces, the optional heterocycle aliphatic group that replaces, the optional aryl that replaces, or the optional heteroaryl that replaces.In some embodiments, R ²¹It is the optional alkyl-carbonyl that replaces.

Part comprises its whole stereospecific enantiomers, for example

(when A and B are CH ₂The time, Y ¹And Y ²All be H).

In some embodiments, R ²¹Be the aminoalkyl group carbonyl, halogenated alkyl carbonyl, aromatic yl alkyl carbonyl, aromatic yl alkyl carbonyl, alicyclic alkyl-carbonyl, or assorted alicyclic alkyl-carbonyl, it is optional separately by 1-3 substituting group replacement.In some embodiments, R ²¹It is Heterocyclylalkyl-oxygen base carbonylamino-alkyl-carbonyl, heteroaryl-carbonylamino-alkyl-carbonylamino-alkyl-carbonyl, bicyclic aryl-sulfonamido-alkyl-carbonyl, aryl-alkoxyl group-carbonylamino-alkyl-carbonyl, alkyl-carbonylamino-alkyl-carbonyl, aliphatic group-oxygen base carbonylamino-alkyl-carbonyl, alicyclic group-alkyl-amino carbonyl amino-alkyl-carbonyl, heteroaryl-carbonylamino-alkyl-carbonylamino-alkyl-carbonyl, alkyl-amino carbonyl amino-alkyl-carbonyl, or bicyclic aryl-amino carbonyl amino-alkyl-carbonyl, optional separately by 1-3 substituting group replacement.In some embodiments, R ²²Be the optional aliphatic group that replaces, the optional assorted aliphatic group that replaces, the optional alicyclic group that replaces, the optional assorted alicyclic group that replaces, the optional aryl that replaces or the optional heteroaryl that replaces.In some embodiments, R ²²Be the optional phenyl that replaces, the optional naphthyl that replaces, the optional anthryl that replaces, the optional naphthalene that replaces or the optional anthracene that replaces.In some embodiments, X ¹, X ², Y ¹And Y ²Be respectively hydrogen, a and b are respectively 1.

In some embodiments, R ²¹It is the optional alkyl-carbonyl that replaces.

In some embodiments, R ²¹Be the aminoalkyl group carbonyl, halogenated alkyl carbonyl, aromatic alkyl carbonyl, aromatic alkyl carbonyl, alicyclic alkyl carbonyl or heterolipid naphthene base carbonyl, it is optional separately by 1-3 substituting group replacement.

In some embodiments, R ²¹It is Heterocyclylalkyl-oxygen base carbonylamino-alkyl-carbonyl, heteroaryl-carbonylamino-alkyl-carbonylamino-alkyl-carbonyl, bicyclic aryl-sulfonamido-alkyl-carbonyl, aryl-alkoxyl group-carbonylamino-alkyl-carbonyl, alkyl-carbonylamino-alkyl-carbonyl, aliphatic group-oxygen base carbonylamino-alkyl-carbonyl, alicyclic group-alkyl-amino carbonyl amino-alkyl-carbonyl, alicyclic group-alkyl-carbonylamino-alkyl-carbonyl, heteroaryl-carbonylamino-alkyl-carbonylamino-alkyl-carbonyl, alkyl-amino carbonyl amino-alkyl-carbonyl, or bicyclic aryl-amino carbonyl amino-alkyl-carbonyl, optional separately by 1-3 substituting group replacement.

In some embodiments, R ²²Be the optional aliphatic group that replaces, the optional assorted aliphatic group that replaces, the optional alicyclic group that replaces, the optional assorted alicyclic group that replaces, the optional aryl that replaces or the optional heteroaryl that replaces.

In some embodiments, R ²²Be the optional phenyl that replaces, the optional naphthyl that replaces, the optional anthryl that replaces, the optional naphthalene that replaces or the optional anthracene that replaces.

In some embodiments, X ¹, X ², Y ¹And Y ²Be respectively hydrogen, a and b are respectively 1.

In some embodiments, R ²²Be the optional aliphatic group that replaces, the optional assorted aliphatic group that replaces, the optional alicyclic group that replaces, the optional assorted alicyclic group that replaces, the optional aryl that replaces or the optional heteroaryl that replaces.

In one embodiment, compound is

The deuterated counterpart is not slow than it for the epimerization of deuterate compound of the present invention.As shown below, deuterated compound 1 is converted into not deuterated intermediate very slowly, is converted into epimer 2 and 3 then.Epimer 2 and 3 keeps balance then, and this balance has further delayed the epimerization of deuterate compound 1.

Because its epimerization is slow, deuterate compound of the present invention with respect to its not the deuterated analogue can improve the bulk concentration of active isomer.

In some embodiments, deuterium enriched amount is at least 50% in the The compounds of this invention.In some embodiments, deuterium enriched amount is at least 80% in the The compounds of this invention.In some embodiments, deuterium enriched amount is at least 90% in the The compounds of this invention.In some embodiments, deuterium enriched amount is at least 99% in the The compounds of this invention.

The invention still further relates to a kind of pharmaceutical composition, it comprises pharmaceutically acceptable carrier and the compound of formula (I) or the compound of the above arbitrary embodiment.

The invention still further relates to the method that increases medicament active isomer bulk concentration, comprise that patient to needs uses the deuterate isomer of this medicament that is enough to produce pharmaceutically-active amount.

The invention still further relates to the method that increases the compound bioavailability, comprise with D atom and replace in the compound hydrogen atom with three-dimensional carbon atom bonding.In one embodiment, the deuterated compound is the compound of formula (I) or the above arbitrary embodiment.

The invention still further relates to the method that suppresses HCV proteolytic enzyme, comprise with deuterated formula (I) or above arbitrary compound of stating embodiment contacting HCV proteolytic enzyme.

The invention still further relates to the patient's who treats the illness of suffering from HCV infection or the mediation of HCV proteolytic enzyme method, comprise to the patient and use the deuterated formula (I) of pharmacy effective dose or the compound of the above arbitrary embodiment.

The method for preparing optically enriched formula 1 compound also within the scope of the invention, wherein

The carbon atom of α and β carboxyl is a stereocenter;

R ₁Be H independently, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces;

R ' ₁It is deuterium;

R ' ₂Be-NHR ₂Or-OE;

R ₂Be H, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces; With

E is C _1-6Alkyl or phenmethyl;

The method comprising the steps of:

A) form formula 1 compound salt and

B) the described salt of crystallization obtains the compound of enantiomeric excess more than 55%.

In some embodiments, R ₁Be C _1-6Alkyl, R ' ₂Be-NHR ₂, R wherein ₂Be C _1-6Alkyl or C _1-6Cycloalkyl.In some embodiments, R ₁Be propyl group and R ₂It is cyclopropyl.

In some embodiments, this method further comprises with amination reagent formula ii compound

Amination is to obtain the compound of formula iii

Still in some embodiments, amination reagent is a trinitride salt, makes the reduction of intermediate triazo-compound by hydrogenation.

In some embodiments, this method further comprises with the undersaturated formula i compound of oxidising agent oxidation, wherein R ' ₂Be-NHR ₂Or-OE, wherein E is C _1-5Alkyl or the optional phenmethyl that replaces,

To obtain the compound of formula ii.

In some further embodiments, oxidising agent comprises tertbutyl peroxide.In some further embodiments, oxidising agent further comprises chiral reagent.In some further embodiments, oxidising agent is different third Samarium trioxide (III), triphen arsine oxide, the mixture of S-(-) 1,1 '-two-beta naphthal and 4A molecular sieve.In some further embodiments, oxidising agent is included in the perhydrit under the situation that trifluoroacetic anhydride exists.

In some embodiments, this method comprises that further the compound of hydrolyzing type ii to obtain acid, is converted into acid the amide compound of formula ii, wherein R ' then ₂Be-NHR ₂

The method of preparation formula 1 compound also within the scope of the invention,

Wherein:

R ₁Be H, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces;

R ' ₁It is deuterium;

R ₂Be H, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces; With

Formula I compound enantiomeric excess is more than 55%.

The method comprising the steps of:

A) compound of the undersaturated formula i of oxidation

To obtain the compound of formula ii;

B) compound of formula ii reacts with amination reagent

Obtain the compound of formula iii;

C) form formula iii compound and optical activity organic acid salt;

D) the described salt of crystallization obtains enantiomeric excess and is the compound more than 55%.

In some embodiments, the compound of formula 1 be (2S, 3S)-3-amino-3-deuterium-N-cyclopropyl-2-hydroxyl hexanamide.In some embodiments, organic acid is L-tartrate or Septochol.

The method for preparing optically enriched formula 1 compound also within the scope of the invention,

Wherein:

The carbon atom of α and β carboxyl is a stereocenter;

R ₁Be H independently, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces;

R ' ₁Be deuterium, deuterium enriched amount is at least 50%;

R ' ₂Be-NHR ₂Or-OE;

R ₂Be H, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces; With

E is C _1-6Alkyl or phenmethyl.

The method comprising the steps of: a) form the salt of formula I compound, and b) the described salt of crystallization obtains the compound of enantiomeric excess more than 55%.

In some embodiments, R ₁Be C _1-6Alkyl, R ' ₂Be-NHR ₂, R wherein ₂Be C _1-6Alkyl or C _1-6Cycloalkyl.In some embodiments, R ₁Be propyl group and R ₂It is cyclopropyl.

In some embodiments, this method further comprises with amination reagent formula ii compound

Amination is with the step of the compound that obtains formula iii

In some embodiments, amination reagent is a trinitride salt, makes the reduction of intermediate triazo-compound by hydrogenation.

In some embodiments, this method further comprises with the undersaturated formula i compound of oxidising agent oxidation, wherein R ' ₂Be-NHR ₂Or-OE, wherein E is C _1-5Alkyl or the optional phenmethyl that replaces,

To obtain the compound of formula ii.

In some further embodiments, oxidising agent comprises tertbutyl peroxide.In some further embodiments, oxidising agent further comprises chiral reagent.In some further embodiments, oxidising agent is different third Samarium trioxide (III), triphen arsine oxide, the mixture of S-(-) 1,1 '-two-beta naphthal and 4A molecular sieve.In some further embodiments, oxidising agent is included in the perhydrit under the situation that trifluoroacetic anhydride exists.

In some embodiments, R ' ₂Be-OE.In some embodiments, R ' ₂Be-NHR ₂

In some embodiments, this method comprises that further the compound of hydrolyzing type ii to obtain acid, is converted into acid the amide compound of formula ii, wherein R ' then ₂Be-NHR ₂

In some embodiments, this method further comprises the compound of oxidation-type iv

To obtain the compound of formula ii.In some cases, oxidation is undertaken by using Manganse Dioxide.

In some embodiments, this method further comprises the compound of reduction-type V

To obtain the compound of formula iv.In some cases, use

Reducing compound is used the water-d2 cancellation then.As known in the art, "

" refer to compound [(CH ₃OCH ₂OCH ₂) ₂AlH ₂] Na, its commercially available acquisition is generally toluene solution (for example, 70%W/W).For

More information, can be referring to people such as for example Bates R.W, Tetrahedron, 1990,46,4063.

The method of preparation I compound also within the scope of the invention,

Wherein:

R ₁Be H, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces;

R ' ₁It is deuterium;

R ₂Be H, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces; With

Formula I compound enantiomeric excess is more than 55%.

The method comprising the steps of:

A) compound of the undersaturated formula I of oxidation

To obtain the compound of formula ii;

B) compound with formula ii reacts with amination reagent

To obtain the compound of formula iii;

C) form formula iii compound and optical activity organic acid salt;

D) the described salt of crystallization obtains enantiomeric excess and is the compound more than 55%.

In some embodiments, the compound of formula I be (2S, 3S)-3-amino-3-deuterium-N-cyclopropyl-2-hydroxyl hexanamide.In some embodiments, organic acid is L-tartrate or Septochol.

Detailed Description Of The Invention

I. definition

A. term

As used herein, term " aliphatic group " comprises alkyl, thiazolinyl and alkynyl.

As used herein, term " alkyl " group refers to contain the representative examples of saturated aliphatic alkyl of 1-8 (for example 1-6 or 1-4) carbon atom.Alkyl group can be a straight or branched.The example of alkyl group includes but not limited to methyl, ethyl, propyl group, sec.-propyl, butyl, isobutyl-, sec-butyl, the tertiary butyl, n-pentyl, n-hexyl and 2-ethylhexyl.Alkyl group can be chosen wantonly by one or more substituting groups and replace, and described substituting group is for example alkoxyl group; cycloalkyloxy; the heterocycle alkoxyl group; aryloxy; heteroaryloxy; aralkoxy; assorted aralkoxy; amino; nitro; carboxyl; cyano group; halo; hydroxyl; sulfo-; sulfydryl; alkyl alkylthio base (alkylsulfanyl); alkyl sulphinyl (alkylsulfinyl); alkyl sulphonyl; aminocarboxyl; alkyl-carbonyl-amino; cycloalkyl amino carbonyl; cycloalkyl alkyl carbonyl amino; aryl-amino-carbonyl; aromatic alkyl carbonyl amino; Heterocyclylalkyl-carbonylamino; Heterocyclylalkyl-alkyl-carbonyl-amino; heteroaryl carbonylamino or heteroaralkyl carbonylamino; urea; thiocarbamide; sulfamyl; sulphonamide; alkoxyl group glycosyl or alkyl-carbonyl oxygen base.

As used herein, " thiazolinyl " group refers to contain the aliphatic carbons group of 2-8 (for example, 2-6 or 2-4) carbon atom and at least one two key.Similar to alkyl group, alkenyl group can be straight or branched.The example of alkenyl group includes but not limited to allyl group, prenyl, crotyl and 2-hexenyl.Alkenyl group can be chosen wantonly by one or more substituting groups and replace, and described substituting group is for example alkoxyl group; cycloalkyloxy; the heterocycle alkoxyl group; aryloxy; heteroaryloxy; aralkoxy; assorted aralkoxy; amino; nitro; carboxyl; cyano group; halo; hydroxyl; sulfo-; sulfydryl; alkyl alkylthio base; alkyl sulphinyl; alkyl sulphonyl; aminocarboxyl; alkyl-carbonyl-amino; cycloalkyl amino carbonyl; cycloalkyl-alkyl-carbonyl-amino; aryl-amino-carbonyl; aromatic alkyl carbonyl amino; Heterocyclylalkyl-carbonylamino; Heterocyclylalkyl-alkyl-carbonyl-amino; the heteroaryl carbonylamino; the heteroaralkyl carbonylamino; urea; thiocarbamide; sulfamyl; sulphonamide; alkoxyl group glycosyl or alkyl-carbonyl oxygen base.

As used herein, " alkynyl " group refers to contain 2-8 (for example, 2-6 or 2-4) carbon atom and at least one triple-linked aliphatic carbons group.Alkynyl group can be straight or branched.The example of alkynyl group includes but not limited to propargyl, butynyl.Alkynyl group can be chosen wantonly by one or more substituting groups and replace, and described substituting group is for example alkoxyl group; cycloalkyloxy; the heterocycle alkoxyl group; aryloxy; heteroaryloxy; aralkoxy; assorted aralkoxy; amino; nitro; carboxyl; cyano group; halo; hydroxyl; sulfo-; sulfydryl; alkyl alkylthio base; alkyl sulphinyl; alkyl sulphonyl; aminocarboxyl; alkyl-carbonyl-amino; cycloalkyl amino carbonyl; cycloalkyl-alkyl-carbonyl-amino; aryl-amino-carbonyl; aromatic alkyl carbonyl amino; Heterocyclylalkyl-carbonylamino; Heterocyclylalkyl-alkyl-carbonyl-amino; the heteroaryl carbonylamino; the heteroaralkyl carbonylamino; urea; thiocarbamide; sulfamyl; sulphonamide; alkoxyl group glycosyl or alkyl-carbonyl oxygen base.

As used herein, " amino " group refers to-NR ^XR ^Y, each R wherein ^XAnd R ^YBe hydrogen, alkyl, cycloalkyl, (cycloalkyl) alkyl, aryl, aralkyl, Heterocyclylalkyl, (Heterocyclylalkyl) alkyl, heteroaryl or heteroaralkyl independently.When term " amino " was not end group (for example alkyl-carbonyl-amino), it was by-NR ^X-expression.R ^XImplication is identical with above definition.

As used herein, " aryl " group refers to phenyl, naphthyl or has the benzo-fused group of 2 to 3 rings.For example benzo-fused group comprises and one or two C _4-8Alicyclic moiety condensed benzene, for example 1,2,3,4-tetralyl, indanyl, dihydro indanyl or fluorenyl.Aryl can be chosen wantonly by one or more substituting groups and replace; described substituting group (comprises carboxyalkyl for for example alkyl; hydroxyalkyl and haloalkyl, for example trifluoromethyl); thiazolinyl; alkynyl; cycloalkyl; (cycloalkyl) alkyl; Heterocyclylalkyl; (Heterocyclylalkyl) alkyl; aryl; heteroaryl; alkoxyl group; cycloalkyloxy; Heterocyclylalkyl oxygen base; aryloxy; heteroaryloxy; aralkyl oxy; heteroarylalkyl oxygen base; aroyl; 4-hetaroylpyrazol; amino; nitro; carboxyl; alkoxy carbonyl; the alkyl-carbonyl oxygen base; aminocarboxyl; alkyl-carbonyl-amino; cycloalkyl amino carbonyl; (cycloalkyl) alkyl-carbonyl-amino; aryl-amino-carbonyl; aromatic alkyl carbonyl amino; (Heterocyclylalkyl) carbonylamino; (Heterocyclylalkyl) alkyl-carbonyl-amino; the heteroaryl carbonylamino; the heteroaralkyl carbonylamino; cyano group; halo; hydroxyl; acyl group; sulfydryl; alkyl alkylthio base; sulphur oxygen base; urea; thiocarbamide; sulfamyl; sulphonamide; oxo or formamyl.

As used herein, the alkyl group that " aralkyl " group refers to be replaced by aromatic yl group (C for example _1-4Alkyl group)." alkyl " and " aryl " as defined herein.The example of aromatic alkyl group is a phenmethyl.

As used herein, alicyclic radical comprises cycloalkyl, cycloalkenyl group and cycloalkynyl radical.

As used herein, " cycloalkyl " group refers to have 3-10 (for example 4-8 carbon atom) aliphatic carbocycle.The example of group of naphthene base comprises cyclopropyl, cyclopentyl, cyclohexyl, suberyl, adamantyl, norbornene (norbornyl), cube alkyl (cubyl), octahydro indenyl, decahydro naphthyl, dicyclo [3.2.1] octyl group, dicyclo [2.2.2] octyl group, dicyclo [3.3.1] nonyl and dicyclo [3.3.2.] nonyl.As used herein, " cycloalkenyl group " group refers to have the non-aromatic carbocyclic ring with the individual carbon atom of 3-10 (for example 4-8) of one or more pairs of keys.The example of cycloalkenyl groups comprises cyclopentenyl, 1,4-hexamethylene two-alkene-Ji, cycloheptenyl, cyclooctene base, six hydrogen indenyls, octahydro naphthyl, dicyclo [2.2.2] octenyl and dicyclo [3.3.1] nonene base.Cycloalkyl or cycloalkenyl groups can be chosen wantonly by one or more substituting groups and replace; described substituting group (comprises carboxyalkyl for for example alkyl; hydroxyalkyl and haloalkyl, for example trifluoromethyl); thiazolinyl; alkynyl; cycloalkyl; (cycloalkyl) alkyl; Heterocyclylalkyl; (Heterocyclylalkyl) alkyl; aryl; heteroaryl; alkoxyl group; cycloalkyloxy; the heterocycle alkoxyl group; aryloxy; heteroaryloxy; aralkoxy; assorted aralkoxy; aroyl; 4-hetaroylpyrazol; amino; nitro; carboxyl; alkoxy carbonyl; the alkyl-carbonyl oxygen base; aminocarboxyl; alkyl-carbonyl-amino; cycloalkyl amino carbonyl; (cycloalkyl) alkyl-carbonyl-amino; aryl-amino-carbonyl; aromatic alkyl carbonyl amino; (Heterocyclylalkyl) carbonylamino; (Heterocyclylalkyl) alkyl-carbonyl-amino; the heteroaryl carbonylamino; the heteroaralkyl carbonylamino; cyano group; halo; hydroxyl; acyl group; sulfydryl; alkyl alkylthio base; sulphur oxygen base; urea; thiocarbamide; sulfamyl; sulphonamide; oxo or formamyl.

As used herein, the heterolipid cyclic group refers to Heterocyclylalkyl, heterocycloalkenyl and heterocycle alkynyl.As used herein, " Heterocyclylalkyl " group is meant that () saturated rings structure for example, 4 to 8-units, wherein one or more annular atomses are the heteroatomss such as N, O or S in 3 to 10-units.The example of Heterocyclylalkyl is piperidyl, piperazinyl, THP trtrahydropyranyl, tetrahydrofuran base, dioxolanyl, oxazolidinyl, isoxazole alkyl, morpholinyl, octahydro benzofuryl, octahydro benzopyranyl, octahydro benzo thiapyran base, octahydro indyl, octahydro indyl, decahydroquinolyl, octahydro benzo [b] thienyl, 2-oxa--dicyclo [2.2.2] octyl group, 1-aza-bicyclo [2.2.2] octyl group, 3-aza-bicyclo [3.2.1] octyl group and 2,6-two oxa-s-three ring [3.3.1.03,7] nonyl.As used herein, " heterocycloalkenyl " group is meant 3 to 10-units (for example, 4 to 8-units) the non-aromatic ring structure with one or more pairs of keys, and wherein one or more annular atomses are the heteroatomss such as N, O or S.Heterocyclylalkyl or heterocycloalkenyl group can be chosen wantonly by one or more substituting groups and replace; described substituting group is for for example: alkyl (comprises carboxyalkyl; hydroxyalkyl and haloalkyl, for example trifluoromethyl); thiazolinyl; alkynyl; cycloalkyl; (cycloalkyl) alkyl; Heterocyclylalkyl; (Heterocyclylalkyl) alkyl; aryl; heteroaryl; alkoxyl group; cycloalkyl oxy; Heterocyclylalkyl oxygen base; aryloxy; heteroaryloxy; aralkyl oxy; heteroaralkyl oxygen base; aroyl; 4-hetaroylpyrazol; amino; nitro; carboxyl; alkoxy carbonyl; the alkyl-carbonyl oxygen base; aminocarboxyl; alkyl-carbonyl-amino; cycloalkyl amino carbonyl; (cycloalkyl) alkyl-carbonyl-amino; aryl-amino-carbonyl; aromatic alkyl carbonyl amino; (Heterocyclylalkyl) carbonylamino; (Heterocyclylalkyl) alkyl-carbonyl-amino; the heteroaryl carbonylamino; the heteroaralkyl carbonylamino; cyano group; halogen; hydroxyl; acyl group; sulfydryl; alkyl alkylthio base; sulphur oxygen base; urea; thiocarbamide; sulfamyl; sulfoamido; oxo or formamyl.In some cases, Heterocyclylalkyl or heterocycloalkenyl originally on one's body substituting group can be ring-type (optional contain one or more heteroatomss), so that resulting substituted heterocycle alkyl or heterocycloalkenyl are spiro system, for example

Or

As used herein, " heteroaryl " group is meant the monocyclic, bicyclic or tricyclic structure with 5-15 annular atoms, wherein one or more annular atomses be such as the heteroatoms of N, O, S or B and wherein two the ring or tricyclic structure in one or more rings are aromatic rings.Some examples of heteroaryl are pyridyl, furyl, pyrryl, thienyl, thiazolyl, oxazolyl, imidazolyl, indyl, 2,3-indolinyl, quinolyl, 1,2-dihydroquinoline base, 1,2,3,4-tetrahydric quinoline group, tetrazyl, benzofuryl, 2,3-dihydro benzo furyl, benzothiazolyl, xanthene, thioxanthene, thiodiphenylamine, indoline and benzo [1,3] dioxole.Heteroaryl can be chosen wantonly by one or more substituting groups and replace; described substituting group (comprises carboxyalkyl for for example alkyl; hydroxyalkyl and haloalkyl, for example trifluoromethyl); thiazolinyl; alkynyl; cycloalkyl; (cycloalkyl) alkyl; Heterocyclylalkyl; (Heterocyclylalkyl) alkyl; aryl; heteroaryl; alkoxyl group; cycloalkyl oxy; Heterocyclylalkyl oxygen base; aryloxy; heteroaryloxy; aralkyl oxy; heteroaralkyl oxygen base; aroyl; 4-hetaroylpyrazol; amino; nitro; carboxyl; alkoxy carbonyl; the alkyl-carbonyl oxygen base; aminocarboxyl; alkyl-carbonyl-amino; cycloalkyl amino carbonyl; (cycloalkyl) alkyl-carbonyl-amino; aryl-amino-carbonyl; aromatic alkyl carbonyl amino; (Heterocyclylalkyl) carbonylamino; (Heterocyclylalkyl) alkyl-carbonyl-amino; the heteroaryl carbonylamino; the heteroaralkyl carbonylamino; cyano group; halogen; hydroxyl; acyl group; sulfydryl; alkyl alkylthio base; sulphur oxygen base; urea; thiocarbamide; sulfamyl; sulfoamido; oxo or formamyl.

As used herein, " heteroaralkyl " group is meant alkyl-alkyl (for example, the C that is replaced by heteroaryl groups _1-4Alkyl group)." alkyl " and " heteroaryl " as above defines.

As used herein, " circular part " comprises cycloalkyl, Heterocyclylalkyl, cycloalkenyl group, heterocycloalkenyl, aryl or heteroaryl, and each before defined freely.

As used herein, " acyl group " group be meant formyl radical or alkyl-C (=O)-, wherein " alkyl " is as previous definition.The example of carboxyl groups is ethanoyl and valeryl.

As used herein, " formamyl " group is meant to have-O-CO-NR ^XR ^YOr-NR ^X-CO-O-R ^ZKnot

The group of structure, wherein R ^XAnd R ^YAs above definition, R ^ZBe alkyl, aryl, aralkyl, Heterocyclylalkyl, heteroaryl or heteroaralkyl.

As used herein, " carboxyl " and " sulfo group " group refers to respectively-COOH and-SO ₃H.

As used herein, " alkoxyl group " group is meant alkyl-O-group, and wherein " alkyl " is as previous definition.

As used herein, " sulphur oxygen base " is meant-O-SO-R ^XOr-SO-O-R ^X, R wherein ^XAs above definition.As used herein, " sulfane base " refers to-S-R ^X, R wherein ^XAs above definition.

As used herein, " sulfinyl " refers to-S (O)-R ^X, R wherein ^XHas definition above.

As used herein, " alkylsulfonyl " refers to-S (O) ₂-R ^X, R wherein ^XHas definition above.

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

As used herein, " sulfamyl " group refers to structure-S (O) ₂-NR ^xR ^yOr-NR ^x-S (O) ₂-R ^z, R wherein ^x, R ^yAnd R ^zDefinition as mentioned.

As used herein, " sulphonamide " group refers to structure-NR ^X-S (O) ₂-NR ^xR ^y, R wherein ^x, R ^yAnd R ^zDefinition as mentioned.

As used herein, " urea " group refers to structure-NR ^x-CO-NR ^yR ^z, " thiocarbamide " group refers to structure-NR ^X-CS-NR ^YR ^Z, R ^x, R ^yAnd R ^zDefinition as mentioned.

As used herein, " guanidine radicals " group refers to structure-N=C (NR ^xR ^y) N (R ^xR ^y), R wherein ^xAnd R ^zAs above definition.

As used herein, " amidino groups " group refers to structure-C (NR ^x) N (R ^xR ^y), R wherein ^xAnd R ^yAs above definition.

As used herein, term " oximido " refers to structure-C=N-OR ^X, R wherein ^XAs above definition.

As used herein, significant quantity is defined as and produces the required amount of result of treatment in being treated the patient, determines according to patient's age, surface-area, body weight and situation usually.Mutual relationship between animal and human's class dosage (based on the milligram quantities of every square metre of body surface area) is by people such as Freireich, Cancer Chemother.Rep., and 50:219 (1966) is described.Body surface area can generally be determined according to patient's height and body weight.Referring to, Scientific Tables for example, GeigyPharmaceuticals, Ardsley, New York, 537 (1970).

As used herein, " patient " refers to comprise the mankind by Mammals.

As used herein, antagonist refers to still can not activate with receptors bind the molecule of this receptor.Binding site on itself and endogenic ligand or the substrate competition acceptor suppresses the ability of signal in acceptor conduction and the endogenic ligand bonded cell thus.

Phrase " optional replacement " can exchange with phrase " replacement or unsubstituted " and use.As described herein, compound of the present invention can be chosen wantonly by one or more substituting groups and replace, described substituting group for for example above exemplified common those, or as an example particular category of the present invention, subclass and kind.Except as otherwise noted, variable R 1 in the formula (I), R ², R ³, R ⁴And R ⁵Each concrete group can choose wantonly by one or more substituting groups as herein described and replace.The substituting group of each concrete group is further optional to be replaced by one to three halogen, cyano group, alkoxyl group, hydroxyl, nitro, haloalkyl and alkyl.For example, alkyl can be replaced by alkyl alkylthio base, and alkyl alkylthio base is optional to be replaced by one to three halogen, oxo, cyano group, alkoxyl group, hydroxyl, nitro, haloalkyl and alkyl.As additional embodiments, alkyl can be replaced by (cycloalkyl) carbonylamino, and the cycloalkyl moiety of (cycloalkyl) carbonylamino is optional to be replaced by one to three halogen, cyano group, hydroxyl, nitro, haloalkyl and alkyl.

Usually, term " replacement ", no matter whether its front has term " randomly ", all refers to replace hydrogen atom group with the atomic group of specified substituent in given structure.Specified substituent defines and as mentioned hereinafter to the description of compound and embodiment.Except as otherwise noted, randomly the group of Qu Daiing has substituting group in the part that respectively can replace of this group, when the more than one substituting group that is selected from special groups when more than one position in any given structure replaced, the substituting group of each position can be identical or different.Ring substituents, for example Heterocyclylalkyl can encircle with another, and for example the cycloalkyl bonding forms the spiral shell bicyclic ring system, and for example, two rings have a common atom.It will be recognized by those skilled in the art that the desired substituent combination of the present invention is to form those combinations stable or compound that chemistry is feasible.

As used herein, phrase " stable or chemistry feasible " refers to preferably experience its recovery, purification condition when its condition of producing, detecting of experience, and when being used for one or more purpose disclosed herein, basically the compound that can not change.In certain embodiments, stable compound or the feasible compound of chemistry be meant at 40 ℃ or low temperature more, lacks and keep the compound that do not change basically at least one week under moisture or other the chemically reactive conditions.

Except as otherwise noted, structure described herein also mean comprise this structure whole isometrys (for example, enantiomer, diastereomer and geometrical isomer (or conformer)) form, the for example R of each asymmetric center and S configuration, (Z) and (E) double bond isomer, and (Z) and (E) conformer.Therefore, the mixture of the independent three-dimensional chemical isomer of The compounds of this invention and enantiomer, diastereomer and geometrical isomer (or conformer) all within the scope of the invention.Except as otherwise noted, whole tautomeric forms of The compounds of this invention within the scope of the invention.

In addition, except as otherwise noted, structure described herein also means and comprises only at the compound that has difference aspect one or more isotopic enrichment atoms.For example, the compound with structure of the present invention is used except that replacing the hydrogen with deuterium or tritium ¹³C or ¹⁴C enrichment carbon atom replaces carbon within the scope of the invention.This compounds is useful analysis tool or probe in biological example is measured.

The N-oxide compound of each formula (I) compound or pharmacy acceptable salt are also within the scope of the invention.For example, imidazoles or pyrazoles center ring or contain the substituent azo-cycle atom of heterocyclic radical of nitrogen can be at the oxygenant that is fit to, for example metachloroperbenzoic acid or H ₂O ₂Form oxide compound under the situation about existing.

Be essentially tart formula (I) compound (having carboxyl or phenolic hydroxyl group group) and can form pharmacy acceptable salt, for example sodium, potassium, calcium or golden salt.With pharmaceutically acceptable amine, for example the salt of ammoniacal liquor, alkylamine, hydroxyalkyl amine and N-methyl glycamine formation also within the scope of the invention.The compound usable acid of formula (I) is handled and is formed acid salt.This type of sour example comprises hydrochloric acid, Hydrogen bromide, hydroiodic acid HI, sulfuric acid, methylsulfonic acid, phosphoric acid, to bromophenyl sulfonic acid, carbonic acid, succsinic acid, citric acid, phenylformic acid, oxalic acid, propanedioic acid, Whitfield's ointment, oxysuccinic acid, fumaric acid, xitix, toxilic acid, acetate and other mineral acids well-known in the art and organic acid.Acid salt can prepare to produce acid salt (for example hydrochloride) by formula (I) compound of handling free alkali form with the acid (for example hydrochloric acid) of capacity.Acid salt can be transformed back its free alkali form by handling described salt with the dilution alkali aqueous solution (for example, sodium hydroxide, sodium bicarbonate, salt of wormwood or ammoniacal liquor) that is fit to.The compound of formula (I) can also be for example to be non-chiral compound, racemic mixture, optically active compound, the diastereomer of purifying or the mixture of diastereomeric compound.

B. abbreviation

Following abbreviation has following implication.If abbreviation is definition not, it has its implication of accepting usually.

BEMP=2-tertiary butyl imido grpup-2-diethylamino-1,3-dimethyl perhydro-1,3,2-diaza phosphorane (phosphorine)

The Boc=tert-butoxycarbonyl

BOP=benzotriazole-1-base oxygen base three (dimethylamino) phosphofluoric acid phosphonium salt

Bd=is wide bimodal

Bs=is wide unimodal

The CDI=carbonyl dimidazoles

D=is bimodal

Bimodal bimodal of dd=

The DIC=DIC

The DMF=dimethyl formamide

The DMAP=dimethyl aminopyridine

The DMSO=dimethyl sulfoxide (DMSO)

EDCI=ethyl-1-(3-dimethylaminopropyl) carbodiimide

The eq=equivalent

The EtOAc=ethyl acetate

The g=gram

The HOBT=1-hydroxybenzotriazole

DIPEA=Hunig ' s alkali=diisopropylethylamine

The L=liter

The m=multiplet

The M=mole

The max=maximum

The meq=milliequivalent

The mg=milligram

The mL=milliliter

The mm=millimeter

The mmol=mmole

MOC=methoxyl group oxygen base carbonyl

N=is normal

N/A=does not obtain

The ng=nanogram

The nm=nanometer

The OD=optical density(OD)

PEPC=1-(3-(1-pyrrolidyl) propyl group)-3-ethyl carbodiimide

PP-HOBT=piperidines-piperidines-I-hydroxybenzotriazole

Psi=pound/square inch

The Ph=phenyl

The q=quartet

The quint.=quintet

The rpm=rotations per minute

S=is unimodal

The t=triplet

The TFA=trifluoroacetic acid

The THF=tetrahydrofuran (THF)

The tlc=thin-layer chromatography

μ L=microlitre

The UV=ultraviolet ray

II compound of the present invention

Usually, deuterate compound of the present invention can by known in the art synthetic they not the method for deuterate form synthesize, except in building-up process, using deuterate starting material or reaction reagent.The example of applicable method comprises and is described in U.S. Patent application the 60/711st, No. 530; WO02/18369; WO 07/022459; Advanced Organic Chemistry, second edition, the 204th page, J.March, McGraw Hill, New York, NY, 1997, and Synthesis of A:Elemesand Ragnarsosson, J.of Chem.Soc, Perkin1, those in 1996,537.

Whole publications that this paper quotes are introduced its full content by reference.

The compound of formula I can use known method preparation, and is for example illustrational at following schema I.

Schema I

When relating to schema I, with formula iAcid and deuterated formula IiAmmonia alcohol-acid amides under the situation that condensation reagent exists reaction so that formula to be provided IiiHydroxyl-acid amides, described condensation reagent is for example EDCI and HOSu.In some embodiments, exist IiMiddle deuterium (D) the enriching quantity percentage ratio that shows is more than 10%.In other embodiments, enriching quantity is 10% to 99.95%, 40% to 99.95%, 50% to 99.95%, 60% to 99.95%, 80% to 99.95%, 90% to 99.95%, 93% to 99.95%, 97% to 99.95% or 99 to 99.95%, or 99.95% or higher.With the oxygenant oxidation that is fit to IiiSo that the compound of formula I to be provided.The oxidising agent that is fit to comprises, for example Dess-Martin periodo alkane (periodinane) or TEMPO and clorox.

The formula that in schema I, shows IiDeuterate amino-alcohol-acid amides can be by using known method preparation, and for example, following schema II illustrational.

Schema II

When relating to schema II, with formula IvGlycine imine (iminic) ester be converted into formula ViiThe deuterate sultam can carry out (Y.Elemes and U.Ragnarsson, J.Chem.Soc, Perkin I, 1996 according to former described method, 6, (people such as L.Lankiewicz, J.Chem.Soc, Perkin I p.537. as discussed previously, the 1994,17,2503rd page), handle formula continuously with bronsted lowry acids and bases bronsted lowry ViiCompound, handle intermediate amino acid (not shown) with the benzyloxy carbonyl chloride subsequently protected deuterate amino acid be provided ViiiUnder the situation that condensation reagent CDI exists, ViiiProvide formula with the reaction of methoxyl group methylamine IxThe Weinreb acid amides.With for example diisobutyl aluminium hydride or lithium aluminium hydride reduction ViAldehyde is provided xUse and previous described those similar methods (referring to, for example, WO02/18369), with aldehyde xBe converted into cyanalcohol Xi, obtain protected hydroxyl-amino acid thus XiiWith acid XiiBe converted into protected acid amides Xiii, with its deprotection so that amino-acid amides to be provided Ii

Alternatively, can prepare deuterate amino-acid amides ii, the wherein R that in schema I, describes ₂Be H, for example, illustrational as schema III institute.

Schema III

When relating to schema III, with two (2-methoxy ethoxy) sodium aluminum hydride reduction propargyl alcohols Xiv, make the reaction mixture cancellation so that the deuterated vinyl carbinol to be provided with water-d2 subsequently XvUse the Manganse Dioxide oxidation XvAldehyde is provided Xvi, under the situation of sodium phosphate and the existence of 2-methyl-2-butene, with Textone (NaClO ₂) it further is oxidized to acid XviiAcid XviiReact under the situation that N-methylmorpholine exists with isobutyl chloroformate (ICBF), with rear center body mixed anhydride and amine HNR ₄R ₅Reaction acid amides is provided XviiiUnder the situation of trifluoroacetic acid and tosic acid existence, use perhydrit (UHP) epoxidation XviiiSo that epoxide to be provided Xix XixProvide the intermediate triazo-compound with the reaction of sodiumazide Xx, it carries under the situation that palladium exists at carbon subsequently and is reduced to racemize ammonia alcohol by catalytic hydrogenation XxiRacemize ammonia alcohol XxiCan prepare optically active derivatives or form salt by using for example chiral chromatography of known method, split by the organic solvent crystallization subsequently with optical activity acid HA.The optical activity organic acid that is suitable for preparing salt comprises; L-tartrate for example; L MALIC ACID; (S)-amygdalic acid; (1S)-(+)-the 10-camphorsulfonic acid; (-) 2; 2:4; 6-two-O-sec.-propyl diene-2-ketone-L-gulonic acid hydrate; N-ethanoyl-L-leucine; Septochol; (+)-O; O '-dibenzoyl-D-tartrate; O; O '-two-(4-toluyl)-D-tartrate; S-(+) 1,1-dinaphthyl-2-2-phosphoric acid; L-lactic acid; the D-glyconic acid; lactobionic acid; dipivalyl-L-tartrate; S-(+)-O-ethanoyl amygdalic acid and S-(-) 2-(phenylamino methanoyl) propionic acid.The example that is used for the suitable organic solvent of recrystallization comprises N,N-DIMETHYLACETAMIDE, ethyl acetate and acetone.

Can be by conventional analytical procedure, for example NMR and mass spectrograph come thus obtained deuterate compound is differentiated.Can use NMR to determine the structure of compound, and mass spectrograph can be used for by with compound and its not the deuterate form relatively come to determine the amount of D atom in the compound.

The deuterated analogue is more unstable than it usually for deuterate compound of the present invention, and the differentiation tendency is littler.Therefore, they needing can be used for the application of the specific steric configuration of The compounds of this invention.For example, the deuterate compound of formula (I) can be used for treating or prevents that HCV from infecting or the illness of other HCV proteolytic enzyme mediations, because they can suppress HCV proteolytic enzyme.Can measure their inhibition activity by conventional enzyme inhibition test, wherein some are described in publication cited above.Referring to, Perni for example, people such as R.B., Antimicrobial Agents and Chemotherapy, 2006 (march), 50 (3): 899-909.

In addition, the deuterate compound of formula (I) can be used as the biology instrument and studies its not pharmacological characteristics of deuterate form.Therefore, these purposes also within the scope of the invention.

Embodiment 1. preparation (1S; 3aR; 6aS)-2-((S)-2-((S)-2-cyclohexyl-2-(pyrazine-2-carboxamido) kharophen)-3,3-dimethyl butyrate acyl group)-N-((S)-1-(cyclopropyl amino)-1,2-dioxo-3-deuterium-oneself-the 3-yl) octahydro ring penta [c] pyrroles-1-carboxylic acid amides

Step a: preparation

By currently known methods, for example be described in Y.Elemes and U.Ragnarsson, J.of Chem.Soc, Perkin1,1996,6,537; People such as W.Oppolzer, Helv.Chim.Acta., 1994, those methods of 25:2363 use corresponding unsubstituted sultam and propyl iodide to prepare deuterated sultam (the i.e. compound that shows in following schema Vi).

Then with the compound of 17.32g Vi(45.8mmol) and 229mL THF pack into and be equipped with magnetic stirring bar and N ₂In the 500mL round-bottomed flask of inlet mouth.Gained solution is cooled to-78 ℃, in 1 hour with syringe pump add n-BuLi (31.5mL, the 1.6M solution in hexane, 50.3mmol).Before adding HPMA (56mL) solution, with gained yellow solution slaking 30 minutes, (13.4mL 137mmol) added wherein with n-Prl in 30 minutes.In 8 hours with mixture heating up to room temperature, in mixture, adding D then ₂Before the O (50mL), mixture is cooled to-20 ℃.Use EtOAc (400mL) extractive reaction mixture then, with organic layer at MgSO ₄Last dry, concentrate and obtain the 61.3g raw oil.Carry out chromatography on 500g silica gel, with 2:1 heptane/EtOAc wash-out, the cutting belt of enrichment method (rich cut) obtains the 20.35g white solid subsequently.Then with white solid from EtOH (210mL) recrystallization, obtain compound into white crystal solid 15.39g ViiBy ¹It is 93% that H NMR measures the deuterium that mixes.

Step b: preparation (S)-2-(benzyloxy carbonylamino)-2-contains deuterium valeric acid, viii

Compound with step a Vii(15.39g 32.1mmol) packs among THF (100mL) and the 1NHCl (50mL).Spend the night in stirring at room gained emulsion, vacuum concentration is to provide heavy-gravity oil then.Oil is dissolved among the THF (100mL) then, in solution, add entry (25mL) and LiOH (3.08g, 128mmol).This solution stirred once more in room temperature spend the night, concentrate then and remove THF.The pale yellow solution that residue is muddy.With its water (25mL) dilution, and use CH ₂Cl ₂Extraction (three times, each 50mL).Water diluted with THF (200mL) and be cooled to 0 ℃, stir fast simultaneously, in 15 minutes, dropwise add CBZ-Cl (7.6mL, 54mmol).After 1 hour, vacuum is removed the THF solvent 0 ℃ of stirring, and residue is by adding 1N HCl (50mL) acidifying.With this solution with EtOAc extraction (3 times, each 100mL), with organic phase at Na ₂SO ₄The last dry and concentrated oil that obtains.Residue is dissolved in EtOAc (25mL) and the heptane (150mL), adds crystal seed, and in stirred overnight at room temperature.Go up the collection solid at frit (frit), with heptane (30mL) drip washing, the air-dry 5.65g (70%) that obtains is at the compound shown in the above schema ViiiBy ¹It is 93% that H NMR measures the deuterium that mixes.

Step c: preparation (S)-phenmethyl 1-(methoxyl group (methyl) amino)-1-oxo-2-deuterium penta-2-aminocarbamic acid ester

To the N-methylmorpholine (700 μ L) that adds 3.0eq in the flask that 1.0g (S)-2-(benzyloxy carbonylamino)-2-in the 20mL methylene dichloride contains deuterium valeric acid (3.97mmol) that contains that remains on 0 ℃, 1.5eq N, 0-dimethyl oxammonium hydrochloride (581mg) and 1.5eq EDCI (1.14g).With reaction mixture from 0 ℃ to stirred overnight at room temperature.Use the methylene dichloride diluted reaction mixture then, with HCl (1N) and salt water washing.With organic layer at MgSO ₄Last dry.Crude mixture obtains the acid amides (title compound) of 814mg purifying by flash chromatography (ethyl acetate of the 15-75% in hexane) purifying.ES+＝296.1，ES-＝295.2。With ¹H NMR spectrum is confirmed its structure.

Steps d: preparation (S)-phenmethyl 1-oxo-2-deuterium penta-2-aminocarbamic acid ester

The method that use is described in WO 02/18369, the amino acid that Cbz-among the step c is protected is converted into title compound.Particularly.(810mg, the lithium borohydride solution (1.0M) that slowly adds 1.7eq in flask 2.75mmol) (4.67mL) to 1.0eq (S)-phenmethyl 1-(methoxyl group (methyl) the amino)-1-oxo-2-deuterium penta-2-aminocarbamic acid ester in the anhydrous THF of 10mL of containing that remains on 0 ℃ (in ice bath).After stirring about 10 minutes, remove ice bath, continue reaction 1 hour.Pass through to add 5mL KHSO at 0 ℃ ₄(10%) solution makes the reaction cancellation.Come diluting soln by adding 10mL HCl (1N) then.Mixture was stirred 30 minutes, use dichloromethane extraction then 3 times.Organic phase is merged, and with HCl (1N) solution, water and salt water washing.Then with organic phase at MgSO ₄Last dry, the evaporation volatile matter.In next step, use this aldehyde.ES+＝237.1，ES-＝235.2。

Step e: the preparation phenmethyl (3S)-1-(cyclopropyl amino)-2-hydroxyl-1-oxo-3-deuterium oneself-3-aminocarbamic acid ester

The cyclopropyl isonitrile prepares according to schema shown below.

Aldehyde product coupling with cyclopropyl isonitrile and steps d obtains title compound then, as people such as J.E.Semple, and Org.Lett., 2000,2 (18), p.2769; Lumma W., J.Org.Chem., 1981,46,3668 " described in.ES+＝322.1。

Step f: preparation (3S)-3-amino-N-cyclopropyl-3-deuterium-2-hydroxyl hexanamide

Under the situation that hydrogen exists, pass through to use carbon-containing palladium catalyst that the Cbz compound hydrogenolysis of step e is obtained title compound.That show in following schema is step c, d, e and f.

Step g: preparation (1S; 3aR; 6aS)-2-((S)-2-((S)-2-cyclohexyl-2-(pyrazine-2-carboxamido) kharophen)-3,3-dimethyl butyrate acyl group)-N-((3S)-1-(cyclopropyl amino)-3-deuterium-2-hydroxyl-1-oxo oneself-3-yl) octahydro ring penta [c] pyrroles-1-carboxylic acid amides

At coupling agent, for example prepare title compound by the hydroxyl glutamine product of step f and the sour condensation that is fit under the situation that EDCI and HOSu exist.Particularly; to (the 1S that contains the 1.2eq in 20mLDMF; 3aR; 6aS)-2-((S)-2-((S)-2-cyclohexyl-2-(pyrazine-2-carboxamido) kharophen)-3; 3-dimethyl butyrate acyl group) diisopropylamine (980 μ L) of adding 2.5eq in the flask of octahydro ring penta [c] pyrroles-1-carboxylic acid (1.59g), the N-hydroxy benzotriazole hydrate (411mg) of 1.2eq and the EDCI (558mg) of 1.3eq.After 15 minutes, in mixture, add (3S)-3-amino-N-cyclopropyl-3-deuterium-2-hydroxyl hydrochloric acid hexanamide (500mg) of 1.0eq in stirring at room.After other 24 hours, reaction mixture is diluted in the 400mL ethyl acetate.With the organic phase of HCl (1N), water, saturated sodium bicarbonate solution, salt solution purging compound, then in the MgSO4 drying.Coming the purification of crude product to obtain the title compound that 1.31g is a white solid by chromatography (the 70-100% ethyl acetate in hexane) on the silicon-dioxide.ES+＝683.6，ES-＝682.2。Use NMR ¹H confirms its structure.

Step h: preparation (1S; 3aR; 6aS)-2-((S)-2-((S)-2-cyclohexyl-2-(pyrazine-2-carboxamido) kharophen)-3,3-dimethyl butyrate acyl group)-N-((S)-1-(cyclopropyl amino)-1,2-dioxo-3-deuterium-oneself-the 3-yl) octahydro ring penta [c] pyrroles-1-carboxylic acid amides

Title compound passes through with suitable oxygenant, and for example the product of Dess Martin periodo alkane or TEMPO and hypochlorite oxidation step g prepares.Particularly; in room temperature to the 1.31g (1S that contains in the 40mL methylene dichloride; 3aR; 6aS)-add 1.06g Dess Martin periodo alkane in the flask of 2-((S)-2-((S)-2-cyclohexyl-2-(pyrazine-2-carboxamido) kharophen)-3,3-dimethyl butyrate acyl group)-N-((3S)-1-(cyclopropyl amino)-3-deuterium-2-hydroxyl-1-oxo oneself-3-yl) octahydro ring penta [c] pyrroles-1-carboxylic acid amides.Stir after 2 hours, add 50mL sodium bisulfite (1N), mixture was stirred 30 minutes.Separate two-phase, wash organic phase twice with water, use the salt water washing again, dry on Na2SO4.Coming the purification of crude product to obtain the title compound that 1.07g is a white solid by chromatography (the 20-100% ethyl acetate in hexane) on the silicon-dioxide.ES+＝681.5，ES-＝680.0。Use ¹H NMR spectrum is confirmed its structure.

Determine that by MS the deuterium that mixes is 93%.Determine that by chirality positive HPLC the ratio of diastereomer is higher than 99%d.e..

The reaction of following schema step display g and h.

Embodiment 2: and preparation (2S, 3S)-3-amino-3-deuterium-N-cyclopropyl-2--hydroxyl hydrochloric acid hexanamide

Schema shown below is for example understood the overall synthetic of title compound.Each step as detailed below.

Step 1: preparation 3-deuterium-(E)-and oneself-2-alkene-1-alcohol

In the 250mL three neck round-bottomed flasks that are equipped with mechanical stirrer and reflux exchanger, add 2-oneself-1-alcohol (10g, 0.1 mole) and THF (100mL, 10 volumes).The gained mixture is cooled to 0 ± 5 ℃, then under nitrogen atmosphere 0 ℃ to 20 ℃ slowly add Red-Al (in toluene 65%, 32mL, 1.6eq).The gained mixture is warmed to 25 ℃, and stirred 5 hours.Reaction mixture is cooled to-5 ± 5 ℃, between 0 ℃ to 15 ℃, dropwise adds D ₂O (8.2g, 4eq).IPAC (50mL, 5 volumes) and saturated NH pack in the gained mixture ₄Cl solution (50mL, 5 volumes).After stirring the mixture 10 minutes, the white solid that forms is leached.Separate organic layer from filtrate, water layer extracts with IPAC (30mL, 3 volumes).Organic layer merges, water (30mL, 3 volumes) washing and at MgSO ₄Last dry, concentrate and obtain the product (compound 2) that 9.8g is colourless oil.Raw product 2 is not done and is further purified and is used for next step.

¹H?NMR(500MHz，CDCl ₃)δ5.66(t，1H，J＝5.0Hz)，4.12(d，2H，J＝5.0Hz)，2.04(t，2H，J＝5.0Hz)，1.38-1.46(m，2H)，0.93(t，3H，J＝5.0Hz)。

Step 2: preparation 3-deuterium-(E)-and oneself-the 2-olefine aldehydr

In room temperature to being equipped with the containing of mechanical stirrer at CH ₂Cl ₂Activatory MnO packs in the 250mL three neck round-bottomed flasks of the 3-deuterium-2-hexenol in (150mL, 15 volumes) (10g, 0.1 mole) ₂(87g, 10eq).After brute force stirs 1 hour, add other a part of MnO ₂(16g 2eq), continues vibration 4 hours.Reaction soln is filtered by Celite pad.Solvent removed in vacuo (25 ℃ 100mmHg) obtain the crude aldehyde product that 8.8g is a light yellow oil (compound 3).This raw product is not further purified and is used for next step.

¹H?NMR(500MHz，CDCl ₃)δ?9.54(d，1H，J＝IOO?Hz)，6.14(s，1H)，2.34(m，2H)，1.55-1.60(m，2H)，1.00(t，3H，J＝5.0Hz)。

Step 3: preparation 3-deuterium-(E)-and oneself-the 2-olefin(e) acid

3-deuterium-2-hexene-1-the aldehyde (10g, 0.1 mole) of in the 500mL three neck round-bottomed flasks that are equipped with mechanical stirrer and reflux exchanger, packing into, uncle-BuOH (90mL, 9 volumes) and 2-methyl-2-butene (30mL, 3 volumes).The NaClO in water (200mL) that in 30 minutes, in gained solution, adds prepared fresh ₂(27.4g, 3eq) and NaH ₂PO ₄(62.9g, solution 4eq).With reaction mixture stirring at room 2 hours.Reaction soln is cooled to 0 ℃, and adds saturated Na ₂SO ₃The aqueous solution becomes colourless up to reaction color.Stop to stir, separate organic layer, with EtOAc (3 volume x3) aqueous layer extracted.Organic layer is merged, and vacuum concentration becomes 3 volumes up to cumulative volume.With 1N NaOH (3 volume x3) extraction gained solution, abandon remaining organic layer.With 6N HCl the water layer that merges is acidified to pH and becomes 1.0.Use CH ₂Cl ₂(3 volume x5) extracts this solution.With the organic layer that merges at MgSO ₄Last dry, concentrate and obtain the product that 8.7g is a white solid (compound 4).

¹H?NMR(500MHz，CDCl ₃)δ5.84(s，1H)，2.23(t，2H，J＝5.0Hz)，1.51-1.55(m，2H)，0.98(t，3H，J＝5.0Hz)。

Step 4: preparation 3-deuterium-(E)-N-cyclopropyl oneself-2-alkene acid amides

In the 250mL three neck round-bottomed flasks that are equipped with mechanical stirrer and reflux exchanger, be enclosed in CH ₂Cl ₂2-hexenoic acid-3d in (100mL, 10 volumes) (10g, 0.09 mole), and IBCF (13g, 1.1eq).Gained solution is cooled to 0 ℃, by controlled temperature 0 ℃ to 20 ℃ slowly add NMM (13.2g, 1.5eq).Then, mixture is heated to room temperature and stirred 1 hour.In gained solution, add cyclopropylamine (5.9g, 1.2eq), with this solution stirring 2 hours.With 1N NaOH (3 volume x2), 1N HCl (3 volume x2) and salt brine solution (3 volume) and water (3 volume) washing reaction mixture.With organic layer at MgSO ₄Last dry, concentrate the raw product that obtains to oil.Raw product is dissolved in the heptane (5 volume), is cooled to-78 ℃ in the time of stirring.The solid of filtering-depositing, drying obtain the product that 8.7g is a white solid (compound 5).

¹H?NMR(500MHz，DMSO)δ7.92(s，1H)，5.78(s，1H)，2.66-2.68(m，1H)，2.08(t，2H，J＝5.0Hz)，1.38-1.42(m，2H)，0.87(t，3H，J＝5.0Hz)，0.63(t，2H，J＝3.0Hz)，0.40(t，2H，J＝3.0Hz)。

Step 5: preparation 3-deuterium-N-cyclopropyl-3-propyl group oxyethane-Z-carboxylic acid amides

0 ℃ in 30 minutes to being equipped with the containing of mechanical stirrer at CH2Cl ₂(E)-N-cyclopropyl in (100mL, 10 volumes) oneself-2-alkene acid amides-3d (being the product of step 4) (10g, 0.06 mole), perhydrit (25g, 4eq) and right-TsOH (12.3g is added in CH in 250mL three neck round-bottomed flasks 1eq) ₂Cl ₂Trifluoroacetic anhydride in (50mL, 5 volumes) (40.9g, 3eq).Reaction mixture is heated to 40 ± 5 ℃, and stirred 3 hours.After being cooled to 0 ℃, make the reaction mixture cancellation, stirred 30 minutes by slow adding 6N NaOH (100mL, 10 volumes).Wash with the organic layer separation and with salt solution (5 volume) and water (5 volume).With the washing organic layer at MgSO ₄Last dry, evaporating solvent obtains the 9.7g epoxy product (being compound 6) for light yellow oil.This raw product is not further purified and is used for next step.

¹H?NMR(500MHz，DMSO)δ8.01(s，1H)，3.09(s，1H)，2.63-2.65(m，1H)，1.39-1.54(m，4H)，0.91(t，3H，J＝5.0Hz)，0.60(t，2H，J＝3.0Hz)，0.45(t，2H，J＝3.0Hz)。

Step 6: preparation 3-nitrine-3-deuterium-N-cyclopropyl-2-hydroxyl hexanamide

To (the 100mL that contains that is equipped with mechanical stirrer and reflux exchanger at MeOH, 10 volumes) epoxide-3d6 (10g in, 0.06 mole) and anhydrous magnesium sulfate (14.1g, and a sodiumazide of adding in 250mL three neck round-bottomed flasks 2.0eq) (15.3g, 4.0eq).With gained mixture heating up to 65 ± 5 ℃ and stirred 5 hours.Reaction mixture is cooled to room temperature and adds IPAC (100mL, 10 volumes), mixture was stirred other 10 minutes.Mixture is passed through Pad filters to remove insoluble salt, and the gained settled solution is concentrated into 3 volumes.In gained solution, add IPAC (170mL, 17 volumes), mixture was stirred other 10 minutes.Once more solution is passed through

Pad filters and to obtain product, and trinitride-3d (compound 7) is the settled solution in IPAC (approximately 200mL), and it is not further purified and is used for next step.

¹H?NMR(500MHz，DMSO)δ7.91(s，1H)，6.00(d，1H，J＝5.0Hz)，4.03(d，1H，J＝5.0Hz)，2.66-2.67(m，1H)，1.30-1.58(m，4H)，0.88(t，3H，J＝5.0Hz)，0.60(t，2H，J＝3.0Hz)，0.48(t，2H，J＝3.0Hz)。

Step 7: preparation 3-amino-3-deuterium-N-cyclopropyl-2-hydroxyl hexanamide

In hydrogenation reactor to be equipped with containing of mechanical stirrer in the 500mL high-pressure hydrogenation device of the trinitride-3d7 in IPAC (200mL, 0.05 mole) that previous step obtains, pack into Pd/C (10%Pd, water 50%, 0.8g).Charge into nitrogen (1.0atm) to gained solution, discharge three times, charge into hydrogen (3.0atm) then and discharge three times.Charge into hydrogen (3atm) to gained solution, stirred 5 hours.After the release hydrogen, use nitrogen purge solution 5 minutes.In gained solution, add MeOH (30ml, 3 volumes), reaction mixture is heated to 50 ± 5 ℃.By Celite pad the reaction mixture filtration is obtained settled solution.By coming separated product up to residue 3 volume solution at 20 ± 5 ℃ of concentrated solutions.By solid collected by filtration, washing (IPAC, 3 volumes), it is white crystal solid title compound (compound 8) that drying obtains 7.7g. ¹H NMR (500MHz, DMSO) δ 7.70 (s, 1H), 5.31 (s, 2H), 3.68 (s, 1H), 2.64-2.66 (m, 1H), 1.10-1.50 (m, 4H), 0.82 (t, 3H, J=5.0Hz), 0.59 (t, 3H, J=3.0Hz), 0.45 (t, 3H, J=3.0Hz).

Step 8: preparation (2S, 3S)-3-amino-3-deuterium-N-cyclopropyl-2-hydroxyl Septochol hexanamide

To being equipped with mechanical stirrer and containing (100mL at THF, racemize (the 2S of the step 7 10v)), 3S)-and the Septochol of packing in the 250mL three neck round-bottomed flasks of 3-amino-3-deuterium-N-cyclopropyl-2-hydroxyl hexanamide (10g, 0.05 mole) (15.7g, 0.75eq).Reaction mixture is heated to 65 ± 5 ℃ and stirred 1 hour.In 1 hour, the gained uniform mixture is cooled to 23 ± 2 ℃, held it in same temperature range 1 hour.By filtering the solid of collecting precipitation, with THF (50mL, 5 volumes) washing, drying obtains the salt compound that 12.4g is a white solid (compound 9).The enantiomeric ratio of product (ER)=2:98.

Step 9: preparation (2S, 3S)-3-amino-3-deuterium-N-cyclopropyl-2-hydroxyl hydrochloric acid hexanamide

In the 250mL three neck round-bottomed flasks that are equipped with mechanical stirrer, add deoxycholate salt and (come from step 8) and 2-propyl alcohol (62mL, 5 volumes).Gained solution is heated to 75 ± 5 ℃, slowly is added in IPA (12mL, 3eq) 5 to the 6N HCl in, powerful simultaneously the stirring.Gained solution was stirred 1 hour in identical temperature, be cooled to 23 ± 2 ℃ then.Reaction mixture was kept 1 hour in identical temperature.By filtering the solid of collecting precipitation, with 2-propyl alcohol washing (36mL, 3 volumes), drying obtains the title compound that 3.0g is a white solid (enantiomeric ratio (ER)=0:100).By MS and ¹H NMR determines that the deuterium that mixes is more than 99%.

¹H?NMR(500MHz，DMSO)δ8.07(s，1H)，7.97(s，3H)，6.25(d，1H，J＝5.0Hz)，4.16(d，1H，，J＝5.0Hz)，2.67-2.70(m，1H)，1.33-1.46(m，4H)，0.84(t，3H，J＝5.0Hz)，0.61(t，3H，J＝3.0Hz)，0.53(t，3H，J＝3.0Hz)。

Embodiment 3: the test of measuring epimerization speed

Deuterated The compounds of this invention epimerization is slow, and is as follows:

According to following experimental measurement epimerization speed.Particularly, 100 μ L substratum (damping fluid, rat plasma, dog plasma or human plasma) are joined in the 96 hole depth plates.In blood plasma, add 10 μ L then and contain test compounds (1S; 3aR; 6aS)-2-((S)-2-((S)-2-cyclohexyl-2-(pyrazine-2-carboxamido) kharophen)-3; 3-dimethyl butyrate acyl group)-N-((S)-1-(cyclopropyl amino)-1; 2-dioxo-3-deuterium-own-3-yl) octahydro ring penta [c] pyrroles's-1-carboxylic acid amides (1 μ M or 10 μ M) acetonitrile solution; simultaneously by using the fluid operated workstation (Hamden of TomTec; CT USA) joins (2mL) in 96 deep-well plates with 1200 μ L ethyl acetate.Then culture plate is closely hidden, with its vortex vibration 20 minutes before centrifugal 10 minutes with 3000rpm.After centrifugal, use TomTec that 900 μ L supernatant are transferred in new V font 96 deep-well plates, then at 20 ℃ under nitrogen (flow velocity 60L/min) dry 30 minutes.Residue is rebuild with 100 μ L, and this solution is transferred to once more in the glass insert of 96 orifice plates.20 μ L are rebuild injection of solution to go among the LC-MS/MS to determine the amount of epimer.The LC-MS/MS spectrometer uses ChiralPak AD post (4.6 x 150mm, 10 μ m), and the mixture of Virahol and normal heptane (10:90,50:50 or 90:10) is a moving phase, and Virahol is a cleaning solvent.The deuterate analogue of use test compound in the MS spectrometer also, described analogue contains 11 D atom (C in cyclohexyl groups ₃₆H ₄₂D ₁₁N ₇O ₆, MW 690.47).

The quality of test compounds (M+H is 681.36 m/z), and its not the quality of deuterate analogue (having identical or different chiral configuration) at the deuteriocarbon center (M+H is 680.36 m/z).Their LC-MS/MS spectrum shows the fragment of 323.30 (having deuterium) and 322.30 (not deuterate).

With two kinds of concentration (1 μ M and 10 μ M) and in identical substratum, (be damping fluid, rat plasma, dog plasma and human plasma), the epimerization speed ratio of deuterated formula (I) compound exhibits of test its deuterate form damping fluid, rat plasma, dog plasma are not slow; Slow many of epimerization speed in human plasma.For example, in human plasma and with 1 μ M or 10 μ M, the deuterate compound is about 30% to isomerization at 180 minutes interpolations, and deuterate form epimerization almost 40% not.In addition, in human plasma, the deuterate compound is with linear velocity epimerization 180 minutes, and initial 60 minutes interpolations being presented at before its horizontal stable of deuterate form are not index speed to isomerization.

Embodiment 4: be determined at IC in the HCV replicon cell ₅₀Test

In this test, use (1S, 3aR, 6aS)-2-((S)-2-((S)-2-cyclohexyl-2-(pyrazine-2-carboxamido) kharophen)-3,3-dimethyl butyrate acyl group)-N-((S)-1-(cyclopropyl amino)-1,2-dioxo-3-deuterium-oneself-the 3-yl) octahydro ring penta [c] pyrroles-1-carboxylic acid amides and (5S, 8S)-3-(5-chloro-2, the 4-Dimethoxyphenyl)-7-((S)-2-(2-cyclohexyl acetyl ammonia)-3,3-dimethyl butyrate acyl group)-N-((S)-1-(cyclopropyl amino)-1,2-dioxo-3-deuterium base-3-yl)-1-oxa--2,7-diaza spiro [4.4] ninth of the ten Heavenly Stems-2-alkene-8-carboxylic acid amides, as Lin, people such as C, J.Biol.Chem., 2004,279:17508-17514; Lin, people such as K, Antimicrob.Agents Chemother., 2004, described in the 48:4784-4792.

The Huh-7 cell that carries self-replicating, subgene group HCV replicon of Conl strain is improved Eagle ' s substratum (DMEM) at Dulbecco ' s, 10% heat inactivation foetal calf serum (FBS), the 2mM L-glutaminate, and nonessential amino acid (JRH Biosciences, Lenexa, KS) and 0.25mg/ml G418 (Invitrogen, Carlsbad, CA) the middle cultivation.The subgene group HCV replicon neomycin phosphotransferase of also encoding, it can make the Huh-7 cell selective growth under the situation that G418 exists with respect to HCV replicon feminine gender of the Huh-7 cell that contains the HCV replicon.Use 4 parametric line matches (SoftMax Pro) in HCV Conl sub-genome duplication daughter cell (19), to measure and make HCV rna level minimizing 50% (IC in the replicon cell ₅₀) or 90% (IC ₉₀), or cell survival reduces by 50% (CC ₅₀) the concentration of test compounds.The replicon cell is contained the test compounds of 2%FBS and .5% DMSO (not having G418) DMEM dilution 37 ℃ of cultivations with usefulness.(QIAGEN, Valencia CA) extract total cell RNA, with copy number quantitative, real-time, multiple reverse transcription-PCR (QRT-PCR, or Taqman) test determination HCV RNA to use the RNeasy-96 test kit.Use is measured the cytotoxicity of compound in HCV replicon cell based on the cell survival test of tetrazolium under identical test is set.

The result shows two kinds of Ki that test compounds had all less than 50nM, IC ₅₀(above 5 days) are less than 10.0 μ M.

Other embodiments

Should be appreciated that though invention has been described in conjunction with detailed specification sheets, the description of preamble is used for explaining, rather than limits the scope of the present invention that is limited by the claims scope.Other aspects, advantage and modification are all within the scope of the appended claims.

Claims (72)

1. the deuterium enriched α-keto acyl ammoniate of following formula

Wherein

D represents D atom;

R ¹Be

Wherein

Be optional monocycle azaheterocyclyl that replaces or the optional many rings azaheterocyclyl that replaces, or the optional many rings azepine heterocycloalkenyl that replaces, wherein unsaturated is away from having R ²¹The ring of part and with-C (O)-N (R ²)-CDR ³-C (O)-C (O)-NR ⁴R ⁵In the ring that part connects;

R ²¹Be Q ³-W ³-Q ²-W ²-Q ¹Wherein

W ²And W ³Be key independently of one another ,-CO-,-CS-,-C (O) N (Q ⁴)-,-CO ₂-,-O-,-N (Q ⁴)-C (O)-N (Q ⁴)-,-N (Q ⁴)-C (S)-N (Q ⁴)-,-OC (O) NQ ⁴-,-S-,-SO-,-SO ₂-,-N (Q ⁴)-,-N (Q ⁴) SO ₂-,-N (Q ⁴) SO ₂N (Q ⁴)-, and work as W ²And W ³When any one was end group, it was a hydrogen;

Q ¹, Q ²And Q ³Be key independently of one another, the optional aliphatic group that replaces, the optional assorted aliphatic group that replaces, the optional cyclic aliphatic base that replaces, the optional aryl that replaces, the optional heteroaryl that replaces, the optional aralkyl that replaces, or the optional heteroaralkyl that replaces; Or work as Q ³, Q ², or Q ¹When any one was end group, it was a hydrogen, and condition is to work as W ³And W ²Q when all existing ²It or not key; With

R ², R ³With R4 be H or C independently of one another _1-6Alkyl; With

R ⁵Be H, alkyl, cycloalkyl, the optional aryl that is replaced by 1-4 alkyl, alkylaryl, aryl, the optional amino that is replaced by 1 or 2 alkyl.

2. the compound of claim 1, wherein R ²¹Be Wherein

R ⁶And R ⁸Be independently of one another

Key; Or

Optional replacement (1,1-or 1,2-) ring alkylidene group; Or

Optional replacement (1,1-or 1,2-) heterocycle alkylidene group; Or

Methylene radical or ethylidene, replaced by a substituting group that is selected from the group of forming by following group, described substituting group is the optional aliphatic group that replaces, optional cyclic group that replaces and the optional aromatic group that replaces, and wherein methylene radical or ethylidene are further optional is replaced by the aliphatic group substituting group;

R ⁷, R ⁹And R ¹¹Be hydrogen or the optional aliphatic group that replaces independently of one another;

R ¹⁰Be the optional aliphatic group that replaces, optional cyclic group that replaces or the optional aromatic group that replaces;

L is-C (O)-,-OC (O)-,-NR ¹¹C (O)-,-S (O) ₂-,-NR ¹¹S (O) ₂-, or key;

N is O or 1.

3. the compound of claim 2, wherein n is 1.

4. the compound of claim 2, wherein R ⁶By a methylene radical that is selected from the substituting group replacement of the group of being made up of following group, described substituting group is the optional aliphatic group that replaces, the cyclic group of optional replacement, the optional aromatic group that replaces.

5. the compound of claim 2, wherein R ⁶The methylene radical that is replaced by isobutyl-.

6. the compound of claim 2, wherein R ⁷Be hydrogen.

7. the compound of claim 2, wherein R ⁸By a methylene radical that is selected from the substituting group replacement of the group of being made up of following group, described substituting group is selected from the optional aliphatic group that replaces, and the optional cyclic group that replaces is chosen the aromatic group that replaces wantonly.

8. the compound of claim 7, wherein R ⁸It is the methylene radical that the cyclic group that is optionally substituted replaces.

9. the compound of claim 8, wherein R ⁸The methylene radical that is replaced by cyclohexyl.

10. the compound of claim 2, wherein R ⁹Be hydrogen.

11. the compound of claim 2, wherein L is-CO-.

12. the compound of claim 2, wherein R ¹⁰It is optional substituted aromatic group.

13. the compound of claim 12, wherein R ¹⁰Be selected from following group:

With

14. the compound of claim 12, wherein R ¹⁰Be the optional pyrazinyl that replaces.

15. the compound of claim 14, wherein R ¹⁰It is the 2-pyrazinyl.

16. the compound of claim 2, wherein It is the monocycle azaheterocyclyl that replaces.

17. the compound of claim 16, wherein

Be the pyrrolidyl that the 3-carbon atom is replaced by heteroaryloxy, wherein heteroaryl is further optional is replaced by 1-4 halogen group.

18. the compound of claim 16, wherein

Be

Or

19. the compound of claim 2, wherein

It is the optional many rings azaheterocyclyl that replaces.

20. the compound of claim 19, wherein

Be

Or

21. the compound of claim 20, wherein

Be

Or

22. the compound of claim 2, wherein R ²Be hydrogen, R ⁴And R ⁵Be hydrogen or cyclopropyl independently of one another.

23. the compound of claim 2, wherein R ³It is propyl group.

24. the compound of claim 2, wherein n is 0.

25. the compound of claim 2, wherein L is-NR ¹¹C (O)-, R ¹¹Be hydrogen.

26. the compound of claim 2, wherein R ¹⁰It is the optional aliphatic group that replaces.

27. the compound of claim 26, wherein R ¹⁰It is the tertiary butyl.

28. the compound of claim 2, wherein this compound is

Or

29. the compound of claim 19, wherein

Be Wherein

Wherein A is-(CHX ¹) _a-;

B is-(CHX ²) _b-;

A is 0 to 3;

B is 0 to 3, and condition is that a+b is 2 or 3;

X ¹And X ²Independently be selected from hydrogen separately, the optional C that replaces _1-4Aliphatic group, the optional aryl that replaces;

Y ¹And Y ²Be hydrogen independently of one another, the optional aliphatic group that replaces, the optional aryl that replaces, amino or-OQ ⁴Q wherein ⁴Be hydrogen or the optional aliphatic group that replaces independently of one another;

R ²²Be the optional aliphatic group that replaces, the optional cyclic aliphatic base that replaces, the optional heterocycle aliphatic group that replaces, the optional aryl that replaces, or the optional heteroaryl that replaces.

30. the compound of claim 29, wherein R ²¹It is the optional alkyl-carbonyl that replaces.

31. the compound of claim 30, wherein R ²¹Be the aminoalkyl group carbonyl, halogenated alkyl carbonyl, aromatic yl alkyl carbonyl, aromatic yl alkyl carbonyl, alicyclic alkyl-carbonyl, or assorted alicyclic alkyl-carbonyl, it is optional separately by 1-3 substituting group replacement.

32. the compound of claim 31, wherein R ²¹It is Heterocyclylalkyl-oxygen base carbonylamino-alkyl-carbonyl, heteroaryl-carbonylamino-alkyl-carbonylamino-alkyl-carbonyl, bicyclic aryl-sulfonamido-alkyl-carbonyl, aryl-alkoxyl group-carbonylamino-alkyl-carbonyl, alkyl-carbonylamino-alkyl-carbonyl, aliphatic group-oxygen base carbonylamino-alkyl-carbonyl, alicyclic group-alkyl-amino carbonyl amino-alkyl-carbonyl, heteroaryl-carbonylamino-alkyl-carbonylamino-alkyl-carbonyl, alkyl-amino carbonyl amino-alkyl-carbonyl, or bicyclic aryl-amino carbonyl amino-alkyl-carbonyl, optional separately by 1-3 substituting group replacement.

33. the compound of claim 29, wherein R ²¹Be Wherein

R ⁶And R ⁸Be independently of one another

Key; Or

Optional replacement (1,1-or 1,2-) ring alkylidene group; Or

Optional replacement (1,1-or 1,2-) heterocycle alkylidene group; Or

Methylene radical or ethylidene, replaced by a substituting group that is selected from the group of forming by following group, described substituting group is the optional aliphatic group that replaces, optional cyclic group that replaces and the optional aromatic group that replaces, and wherein methylene radical or ethylidene are further optional is replaced by the aliphatic group substituting group;

R ⁷, R ⁹And R ¹¹Be hydrogen or the optional aliphatic group that replaces independently of one another;

R ¹⁰Be the optional aliphatic group that replaces, optional cyclic group that replaces or the optional aromatic group that replaces;

L is-C (O)-,-OC (O)-,-NR ¹¹C (O)-,-S (O) ₂-,-NR ¹¹S (O) ₂-, or key;

N is O or 1.

34. the compound of claim 29, wherein R ²²Be the optional aliphatic group that replaces, the optional assorted aliphatic group that replaces, the optional alicyclic group that replaces, the optional assorted alicyclic group that replaces, the optional aryl that replaces or the optional heteroaryl that replaces.

35. the compound of claim 34, wherein R ²²Be the optional phenyl that replaces, the optional naphthyl that replaces, the optional anthryl that replaces, the optional naphthalene that replaces or the optional anthracene that replaces.

36. the compound of claim 29, wherein X ¹, X ², Y ¹And Y ²Be respectively hydrogen, a and b are respectively 1.

37. the compound of claim 36, wherein R ²²Be the optional aliphatic group that replaces, the optional assorted aliphatic group that replaces, the optional alicyclic group that replaces, the optional assorted alicyclic group that replaces, the optional aryl that replaces or the optional heteroaryl that replaces.

38. the compound of claim 37, wherein R ²²Be the optional phenyl that replaces, the optional naphthyl that replaces, the optional anthryl that replaces, the optional naphthalene that replaces or the optional anthracene that replaces.

39. the compound of claim 38, wherein R ²¹It is Heterocyclylalkyl-oxygen base carbonylamino-alkyl-carbonyl, heteroaryl-carbonylamino-alkyl-carbonylamino-alkyl-carbonyl, bicyclic aryl-sulfonamido-alkyl-carbonyl, aryl-alkoxyl group-carbonylamino-alkyl-carbonyl, alkyl-carbonylamino-alkyl-carbonyl, aliphatic group-oxygen base carbonylamino-alkyl-carbonyl, alicyclic group-alkyl-amino carbonyl amino-alkyl-carbonyl, heteroaryl-carbonylamino-alkyl-carbonylamino-alkyl-carbonyl, alkyl-amino carbonyl amino-alkyl-carbonyl, or bicyclic aryl-amino carbonyl amino-alkyl-carbonyl, optional separately by 1-3 substituting group replacement.

40. the compound of claim 29, wherein

Be

41. the compound of claim 40, wherein the structure of this compound is

42. the compound of claim 1, wherein deuterium enriched amount is at least 50% in the compound.

43. the compound of claim 42, wherein deuterium enriched amount is at least 80% in the compound.

44. the compound of claim 43, wherein deuterium enriched amount is at least 90% in the compound.

45. the compound of claim 44, wherein deuterium enriched amount is at least 99% in the compound.

46. a pharmaceutical composition, it comprises pharmaceutically acceptable carrier and claim 1,28,41,44 or 45 each compounds.

47. a method that increases medicament active isomer bulk concentration comprises that patient to needs uses the deuterate isomer of this medicament that is enough to produce pharmaceutically-active amount.

48. the method for claim 47, wherein the deuterate isomer of medicament is a claim 1,28,41,44 or 45 each compounds.

49. a method that increases the compound bioavailability comprises with D atom and replaces in the compound hydrogen atom with three-dimensional carbon atom bonding.

50. the method for claim 49, wherein thus obtained deuterate compound is a claim 1,28,41,44 or 45 each compounds.

51. a method that suppresses HCV proteolytic enzyme comprises with claim 1,28,41,44 or 45 each compound contact HCV proteolytic enzyme.

52. a treatment suffers from, and HCV infects or the patient's of the illness of HCV proteolytic enzyme mediation method, comprises the claim 1 of using pharmacy effective dose to the patient, 28,41,44 or 45 each compounds.

53. a method for preparing optically enriched formula 1 compound, wherein

The carbon atom of α and β carboxyl is a stereocenter;

R ₁Be H independently, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces;

R ' ₁Be that deuterium is so that the enriching quantity of deuterium is at least 50%;

R ₂Be-NHR ₂Or-OE;

R ₂Be H, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces; With

E is C _1-6Alkyl or phenmethyl;

Comprise step:

A) form formula 1 compound salt and

B) the described salt of crystallization obtains the compound of enantiomeric excess more than 55%.

54. the method for claim 53, wherein R ₁Be C _1-6Alkyl, R ' ₂Be-NHR ₂, R wherein ₂Be C _1-6Alkyl or C _1-6Cycloalkyl.

55. the method for claim 54, wherein R ₁Be propyl group and R ₂It is cyclopropyl.

56. the method for claim 53 wherein further comprises with amination reagent formula ii compound

Amination is to obtain the compound of formula iii

57. the method for claim 56, wherein amination reagent is a trinitride salt, makes the reduction of intermediate triazo-compound by hydrogenation.

58. the method for claim 56 wherein further comprises with the undersaturated formula i compound of oxidising agent oxidation, wherein R ' ₂Be-NHR ₂Or-OE, wherein E is C _1-5Alkyl or the optional phenmethyl that replaces,

To obtain the compound of formula ii

59. the method for claim 58, wherein oxidising agent comprises tertbutyl peroxide.

60. the method for claim 59, wherein oxidising agent further comprises chiral reagent.

61. the method for claim 58, wherein oxidising agent is different third Samarium trioxide (III), triphen arsine oxide, the mixture of S-(-) 1,1 '-two-beta naphthal and 4A molecular sieve.

62. the method for claim 58, wherein oxidising agent is included in the perhydrit under the situation that trifluoroacetic anhydride exists.

63. the method for claim 62, wherein R ' ₂Be-OE.

64. the method for claim 62, wherein R ' ₂Be-NHR ₂

65. the method for claim 58, wherein the compound of hydrolyzing type ii is converted into acid the amide compound of formula ii, wherein R ' then to obtain acid ₂Be-NHR ₂

66. the method for claim 58 wherein further comprises the compound of oxidation-type iv

To obtain the compound of formula ii.

67. the method for claim 66, wherein oxidation is undertaken by using Manganse Dioxide.

68. the method for claim 66 further comprises the compound of reduction-type V

To obtain the compound of formula iv.

69. the method for claim 68 is wherein used

Reducing compound is used the water-d2 cancellation then.

70. the method for preparation I compound

Wherein:

R ₁Be H, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces;

R ' ₁It is deuterium;

R ₂Be H, the optional aliphatic group that replaces, the optional alicyclic group that replaces, the optional araliphatic base that replaces, optional assorted aliphatic group that replaces or the optional assorted araliphatic base that replaces; With

Formula I compound enantiomeric excess is more than 55%.

Comprise step:

A) compound of the undersaturated formula i of oxidation

To obtain the compound of formula ii;

B) compound with formula ii reacts with amination reagent

To obtain the compound of formula iii;

C) form formula iii compound and optical activity organic acid salt;

D) the described salt of crystallization obtains enantiomeric excess and is the compound more than 55%.

71. the method for claim 70, the compound of its Chinese style I be (2S, 3S)-3-amino-3-deuterium-N-cyclopropyl-2-hydroxyl hexanamide.

72. the method for claim 71, wherein organic acid is L-tartrate or Septochol.