CN101568336B

CN101568336B - Macrocyclic peptides as hepatitis c virus inhibitors

Info

Publication number: CN101568336B
Application number: CN2007800480628A
Authority: CN
Inventors: A·X·王; B·Z·郑; S·丹德里; N·A·米恩威尔; P·M·斯科拉
Original assignee: Bristol Myers Squibb Co
Current assignee: Bristol Myers Squibb Co
Priority date: 2006-11-16
Filing date: 2007-11-15
Publication date: 2012-07-04
Anticipated expiration: 2027-11-15
Also published as: CN101568336A; ES2356273T3

Abstract

Macrocyclic peptides having the general formula are disclosed. Compositions comprising the compounds and methods for using the compounds to inhibit HCV are also disclosed.

Description

The purposes of Macrocyclic peptides in the preparation hepatitis C virus inhibitors

The cross reference of related application

The application requires the rights and interests of U.S. Provisional Application serial number 60/866,120 (application on November 16th, 2006).

The disclosure is usually directed to antiviral compound; And more specifically relate to the chemical compound that can suppress, comprise the method for this compound compositions and the function that suppresses NS3 protease by NS3 protease (this paper also refers to " the serine protease ") function of hepatitis C virus (HCV) coding.

HCV is main human pathogen, worldwide, estimates to infect 1.7 hundred million people, and be roughly 5 times of 1 type number that HIV infects.The individuality that most of these HCV infect forms serious gradual hepatopathy, comprises liver cirrhosis and hepatocarcinoma.

At present, the most effectively the combination of IFN-and ribavirin is used in the HCV treatment, can in 40% patient, cause lasting effect.Up-to-date clinical effectiveness shows that as the single current system method, the IFN-of PEGization is superior to the IFN-of unmodified.Yet for the experimental therapeutic scheme of the coupling medicine of IFN-that comprises PEGization and ribavirin, most of patient's virus load can not continue to reduce.Thus, the effective therapy for treatment HCV infects also has obvious and unsatisfied needs.

HCV is a positive chain RNA virus.Based on the aminoacid sequence of inferring with in the contrast of the extensive similarity of 5 ' untranslated region, HCV is categorized as the independent kind in the flaviviridae.All members of flaviviridae have the virion of peplos, and this virion contains the positive chain RNA genome, through translating of single successive ORF, and all known virus-specific albumen of this genome encoding.

In nucleotide and in the genomic encoding amino acid sequence of whole HCV, found considerable heterogeneity.Characterize six major gene types, and described the hypotype more than 50.The major gene type distribution worldwide of HCV is different, and the clinical meaning of the genetic heterogeneity of HCV remains unintelligible, although carried out many researchs for genotype in the possible effect aspect pathogenic and the treatment.

The length of strand HCV rna gene group is approximately 9500 nucleotide, and has single ORF (ORF), its about 3000 amino acid whose single big polyproteins of encoding.In infected cell, this polyprotein by cell and virus protease cracking, produces structure and non-structure (NS) property albumen in many sites.With regard to HCV, the generation of adult form non-structural protein (NS2, NS3, NS4A, NS4B, NS5A and NS5B) receives the influence of two kinds of virus proteases.First is in the cracking of NS2-NS3 abutment; Second interior serine protease of N-stub area that is included in NS3; And mediation is in all cracking subsequently in NS3 downstream; Both are with the cis form at the NS3-NS4A cracking site, for remaining NS4A-NS4B NS4B-NS5A, NS5A-NS5B site, are with trans forms.As if NS4A albumen can provide many functions, serves as the cofactor of NS3 protease, and possibly help the film location of NS3 and other rdrp virus component.NS3 albumen and NS4A form complex and are absolutely necessary for effective polyprotein processing, can improve protelytic cracking at all sites.NS3 albumen also demonstrates ribonucleoside triphosphote enzyme and rna helicase enzymatic activity.NS5B is the RNA polymerase that depends on RNA, and it relates in the duplicating of HCV.

The disclosure for example provide with the combination of NS4A protease in can suppress the peptide compounds of the function of NS3 protease.Further, the disclosure has been described and has been given the patient combined therapy, make according to chemical compound of the present disclosure (it can effectively suppress HCV NS3 protease) can with one or two kind of other chemical compound with anti-HCV activity give.

Aspect first, the disclosure provides the chemical compound of formula (I):

Or its officinal salt, wherein

R ¹Be selected from alkoxyl, hydroxyl and-NHSO ₂R ⁷

R ^2aAnd R ^2bBe independently selected from hydrogen and methyl;

R ³Be selected from thiazolinyl, alkyl, aryl, aralkyl, cycloalkyl, (cycloalkyl) alkyl, heterocyclic radical and heterocyclic radical alkyl; Condition is to work as R ⁴When being hydrogen, R then ³It or not heterocyclic radical;

R ⁴Be selected from hydrogen and hydroxyl;

R ⁵Be selected from hydrogen, alkyl and cycloalkyl;

R ⁶Be selected from hydrogen, alkyl, alkoxy carbonyl group, alkylamino radical carbonyl, alkyl-carbonyl, amino carbonyl, aryloxy carbonyl, cycloalkyl oxy carbonyl, di alkylamino group carbonyl, halo alkoxy carbonyl, haloalkyl, halogenated alkyl carbonyl, heterocyclyloxy base carbonyl and (NR ^aR ^b) sulfonyl;

R ⁷Be selected from alkyl, aryl, cycloalkyl, (cycloalkyl) alkyl, heterocyclic radical and-NR ^aR ^bWherein the cycloalkyl moiety of cycloalkyl and (cycloalkyl) alkyl is optional independently is selected from following group replacement by one, two or three: thiazolinyl, alkoxyl, alkoxyalkyl; Alkyl, aralkyl, aryl carbonyl; Cyanic acid, cycloalkenyl group, (cycloalkyl) alkyl; Halogen, halogenated alkoxy, haloalkyl and (NR ^eR ^f) carbonyl; R wherein ^aAnd R ^bBe independently selected from hydrogen, alkoxyl, alkyl, aryl, aralkyl, cycloalkyl, (cycloalkyl) alkyl, haloalkyl, heterocyclic radical and heterocyclic radical alkyl; R wherein ^eAnd R ^fBe independently selected from hydrogen, alkyl, aryl, aralkyl and heterocyclic radical; Wherein the aryl moiety of aryl, aralkyl and heterocyclic radical are optional is replaced by one or two substituent group that is independently selected from alkoxyl, alkyl and halogen; With

Q is C _3-9Saturated or unsaturated chain optionally comprises one to three and is independently selected from O, S (O) _mAnd NR ⁹Hetero atom, wherein m is 0,1 or 2, and R ⁹Be selected from hydrogen, alkoxyl, alkoxy carbonyl group, alkyl, alkyl-carbonyl, alkyl sulphonyl, amino carbonyl, aryl sulfonyl, cycloalkyl, (cycloalkyl) alkyl, cycloalkyl oxy, di alkylamino group carbonyl, di alkylamino group carbonylic alkyl, haloalkyl and heterocyclic radical carbonyl.

In first embodiment aspect first, the disclosure provides the compound or pharmaceutically acceptable salt thereof of formula (I), wherein R ⁴It is hydroxyl.

In second embodiment aspect first, the disclosure provides the compound or pharmaceutically acceptable salt thereof of formula (I), wherein R ¹Be-NHSO ₂R ⁷In the 3rd embodiment aspect first, R ⁷It is cycloalkyl.

In the 4th embodiment aspect first, the disclosure provides the compound or pharmaceutically acceptable salt thereof of formula (I), wherein R ^2aAnd R ^2bBe hydrogen.

In the 5th embodiment aspect first, the disclosure provides the compound or pharmaceutically acceptable salt thereof of formula (I), and wherein Q comprises the heteroatomic C of zero _5-7Unsaturated chain.In the 6th embodiment aspect first, Q comprises the heteroatomic C of zero ₆Unsaturated chain.

Aspect second, the disclosure provides the chemical compound of formula (II):

Or its officinal salt, wherein

R ¹Be-NHSO ₂R ⁷

R ^2aAnd R ^2bBe hydrogen;

R ⁴Be selected from hydrogen and hydroxyl;

R ⁵Be hydrogen;

R ⁶It is alkoxy carbonyl group;

R ⁷Be selected from alkyl, aryl, cycloalkyl, (cycloalkyl) alkyl, heterocyclic radical and-NR ^aR ^bR wherein ^aAnd R ^bBe independently selected from hydrogen, alkoxyl, alkyl, aryl, aralkyl, cycloalkyl, (cycloalkyl) alkyl, haloalkyl, heterocyclic radical and heterocyclic radical alkyl; With

In first embodiment aspect second, the disclosure provides the compound or pharmaceutically acceptable salt thereof of formula (II), wherein R ⁴It is hydroxyl.

In second embodiment aspect second, the disclosure provides the compound or pharmaceutically acceptable salt thereof of formula (II), wherein R ⁷It is cycloalkyl.

In the 3rd embodiment aspect second, the disclosure provides the compound or pharmaceutically acceptable salt thereof of formula (II), and wherein Q comprises the heteroatomic C of zero ₆Unsaturated chain.

Aspect the 3rd, the disclosure provides and has been selected from following chemical compound:

Or its officinal salt.

Aspect the 4th, the disclosure provides the compositions that comprises formula (I) compound or pharmaceutically acceptable salt thereof and pharmaceutically suitable carrier.In first embodiment of fourth aspect, compositions further comprises at least a other chemical compound with anti-HCV activity.In second embodiment aspect the 4th, at least a of other chemical compound is interferon or ribavirin.In the 3rd embodiment aspect the 4th, interferon is selected from the interferon-ALPHA of interferon-ALPHA 2B, PEGization, compound (consensus) interferon, interferon-ALPHA 2A and lymph appearance interferon tau.

In the 4th embodiment aspect the 4th; The disclosure provides compositions; Said composition comprises compound or pharmaceutically acceptable salt thereof, pharmaceutically suitable carrier and at least a other chemical compound with anti-HCV activity of formula (I), wherein other chemical compound at least a be selected from interleukin 2, interleukin 6, interleukin 12, can increase the chemical compound that the response of 1 type helper T lymphocyte forms, interfering RNA, antisense RNA, miaow Kui Mote, ribavirin, inosine 5 '-monophosphate dehydrogenase inhibitor, amantadine and rimantadine.

In the 5th embodiment aspect the 4th; The disclosure provides compositions; Said composition comprises compound or pharmaceutically acceptable salt thereof, pharmaceutically suitable carrier and at least a other chemical compound with anti-HCV activity of formula (I); At least a function that can effectively suppress targeting of other chemical compound wherein; Be used to treat HCV and infect, targeting is selected from HCV metalloproteases, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B albumen, HCV inlet, HCV assembling, HCV outlet, HCV NS5A albumen and IMPDH.

Aspect the 5th, the method that the disclosure provides treatment patient HCV to infect, this method comprises formula (I) compound or pharmaceutically acceptable salt thereof that gives the patient treatment effective dose.In first embodiment aspect the 5th, this method further comprises and gives at least a other chemical compound with anti-HCV activity, and other chemical compound is before giving construction (I) compound or pharmaceutically acceptable salt thereof, give afterwards or with it simultaneously.In second embodiment, at least a in other chemical compound is interferon or ribavirin.In the 3rd embodiment, interferon is selected from interferon-ALPHA, Interferon Alfacon-1, interferon-ALPHA 2A and the lymphoblast interferon tau of interferon-ALPHA 2B, PEGization.

In the 4th embodiment aspect the 5th; The method that the disclosure provides treatment patient HCV to infect; This method comprises formula (I) compound or pharmaceutically acceptable salt thereof and at least a other chemical compound with anti-HCV activity that gives the patient treatment effective dose; Wherein other chemical compound at least a be before giving construction (I) compound or pharmaceutically acceptable salt thereof, give afterwards or with it simultaneously, and at least a other chemical compound is selected from interleukin 2, interleukin 6, interleukin 12, can increase the chemical compound that the response of 1 type helper T lymphocyte forms, interfering RNA, antisense RNA, miaow Kui Mote, ribavirin, inosine 5 '-monophosphate dehydrogenase inhibitor, amantadine and rimantadine.

In the 5th embodiment aspect the 5th; The method that the disclosure provides treatment patient HCV to infect; This method comprises formula (I) compound or pharmaceutically acceptable salt thereof and at least a other chemical compound with anti-HCV activity that gives the patient treatment effective dose; Wherein at least a other chemical compound is before giving construction (I) compound or pharmaceutically acceptable salt thereof, give afterwards or with it simultaneously; Wherein at least a other chemical compound can effectively suppress the function of targeting; HCV infects for treatment, and targeting is selected from HCV metalloproteases, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B albumen, HCV inlet, HCV assembling, HCV outlet, HCV NS5A albumen and IMPDH.

Aspect the 6th, the disclosure provides compositions, its comprise formula (I) compound or pharmaceutically acceptable salt thereof, have a kind of, two kinds, three kinds, four kinds or five kinds of other chemical compounds and pharmaceutically suitable carrier of anti-HCV activity.In first embodiment aspect the 6th, said composition comprises three or four kind of other chemical compound with anti-HCV activity.In second embodiment aspect the 6th, said composition comprises one or two kind of other chemical compound with anti-HCV activity.

Aspect the 7th; The method that the disclosure provides treatment patient HCV to infect; This method comprises formula (I) compound or pharmaceutically acceptable salt thereof that gives the patient treatment effective dose and a kind of, two kinds, three kinds, four kinds or five kinds of other chemical compounds with anti-HCV activity, and other chemical compound is before giving construction (I) compound or pharmaceutically acceptable salt thereof, give afterwards or with it simultaneously.In first embodiment aspect the 7th, this method comprises and gives three or four kind of other chemical compound with anti-HCV activity.In second embodiment aspect the 7th, this method comprises and gives one or two kind of other chemical compound with anti-HCV activity.

Others of the present disclosure can comprise the appropriate combination of embodiment disclosed herein.

Others and embodiment can obtain in the description that this paper provides.

Should be regarded as with law and chemical bonding principle description of the present disclosure consistent.In some cases, possibly remove hydrogen atom, so that hold substituent group at any given position.

Should be appreciated that the chemical compound that the disclosure contained is can suitable those chemical compounds that stably are used as pharmaceutical agent.

All patents, patent application and the list of references quoted in the description are incorporated herein by reference with its integral body in this article.Under inconsistent situation, be as the criterion with the disclosure (comprising definition).

The following term of using in this manual has indicated implication:

The term " thiazolinyl " that this paper uses is meant the straight or branched base of 2 to 6 carbon atoms that contain at least one carbon-to-carbon double bond.

The term " alkoxyl " that this paper uses is meant the alkyl that is connected with parent molecular moiety through oxygen atom.

The term " alkoxyalkyl " that this paper uses is meant by one, two or three substituted alkyl of alkoxyl.

The term " alkoxy carbonyl group " that this paper uses is meant the alkoxyl that is connected with parent molecular moiety through carbonyl.

The term " alkyl " that this paper uses is meant the group derived from the straight or branched saturated hydrocarbons that contains 1 to 6 carbon atom.

The term " alkyl amino " that this paper uses is meant-NHR that wherein R is an alkyl.

The term " alkylamino radical carbonyl " that this paper uses is meant the alkyl amino that is connected with parent molecular moiety through carbonyl.

The term " alkyl-carbonyl " that this paper uses is meant the alkyl that is connected with parent molecular moiety through carbonyl.

The term " alkyl sulphonyl " that this paper uses is meant the alkyl that is connected with parent molecular moiety through sulfonyl.

The term " amino " that this paper uses is meant-NH ₂

The term " amino carbonyl " that this paper uses is meant the amino that is connected with parent molecular moiety through carbonyl.

The term " aryl " that this paper uses is meant phenyl or one of them or two dicyclo condensed ring systems that ring is a phenyl.Dicyclo condensed ring system is by forming with 4 to 6 yuan of fragrance or the condensed phenyl of non-aromatic carbocyclic.Aryl of the present disclosure can be connected with parent molecular moiety through any commutable carbon atom in the group.The representative example of aryl is including, but not limited to indanyl, indenyl, naphthyl, phenyl and tetralyl.Aryl of the present disclosure can be chosen wantonly by one, two, three, four or five and independently be selected from following substituent group replacement: thiazolinyl, alkoxyl, alkoxy carbonyl group, alkyl; Second aryl, aralkyl, aryloxy group, cyanic acid; The cyanic acid alkyl, halogen, halogenated alkoxy, haloalkyl; Heterocyclic radical, heterocyclic radical alkyl, nitro and oxo; Wherein the heterocyclic radical of the aryl moiety of second aryl, aralkyl and aryloxy group, heterocyclic radical and heterocyclic radical alkyl part can further be chosen wantonly by one, two, three, four or five and independently be selected from following substituent group replacement: thiazolinyl; Alkoxyl, alkyl, cyanic acid; Halogen; Halogenated alkoxy, haloalkyl, nitro and oxo.

The term " aralkyl " that this paper uses is meant by one, two or three substituted alkyl of aryl.

The term " aryl carbonyl " that this paper uses is meant the aryl that is connected with parent molecular moiety through carbonyl.

The term " aryloxy group " that this paper uses is meant the aryl that is connected with parent molecular moiety through oxygen atom.

The term " aryloxy carbonyl " that this paper uses is meant the aryloxy group that is connected with parent molecular moiety through carbonyl.

The term " aryl sulfonyl " that this paper uses is meant the aryl that is connected with parent molecular moiety through sulfonyl.

The term " carbonyl " that this paper uses is meant-C (O)-.

The term " cyanic acid " that this paper uses is meant-CN.

The term " cyanic acid alkyl " that this paper uses is meant by one, two or three substituted alkyl of cyanic acid.

The term " cycloalkenyl group " that this paper uses be meant have three to ten four carbon atoms and the heteroatomic non-aromatization of zero, unsaturated monocyclic, dicyclo or the three-loop system of part.The representational example of cycloalkenyl group is including, but not limited to cyclohexenyl group, octahydro naphthyl and norbornene.

The term " cycloalkyl " that this paper uses is meant to have three to ten four carbon atoms and the heteroatomic saturated monocycle of zero, dicyclo or tricyctic hydrocarbon loop systems.The representational example of cycloalkyl is including, but not limited to cyclopropyl, cyclopenta, bicyclo-[3.1.1] heptyl and adamantyl.

The term " cycloalkyl oxy " that this paper uses is meant the cycloalkyl that is connected with parent molecular moiety through oxygen atom.

The term " cycloalkyl oxy carbonyl " that this paper uses is meant the cycloalkyl oxy that is connected with parent molecular moiety through carbonyl.

The term " (cycloalkyl) alkyl " that this paper uses is meant by the alkyl of one, two or three cycloalkyl substituted.

The term " dialkyl amido " that this paper uses is meant-NR ₂, wherein each R is an alkyl.Two R groups can be identical or different.

The term " di alkylamino group carbonyl " that this paper uses is meant the dialkyl amido that is connected with parent molecular moiety through carbonyl.

The term " di alkylamino group carbonylic alkyl " that this paper uses is meant by the alkyl of one, two or three di alkylamino group carbonyl substituted.

Term " halogen " and " halo " that this paper uses are meant F, Cl, Br or I.

The term " halogenated alkoxy " that this paper uses is meant the haloalkyl that is connected with parent molecular moiety through oxygen atom.

The term " halogenated alkoxy alkyl " that this paper uses is meant by one, two or three substituted alkyl of halogenated alkoxy.

The term " halo alkoxy carbonyl " that this paper uses is meant the halogenated alkoxy that is connected with parent molecular moiety through carbonyl.

The term " haloalkyl " that this paper uses is meant by one, two, three or four substituted alkyl of halogen atom.

The term " halogenated alkyl carbonyl " that this paper uses is meant the haloalkyl that is connected with parent molecular moiety through carbonyl.

The term " heterocyclic radical " that this paper uses is meant and contains one, two or three 5,6 or 7 yuan of rings that are independently selected from nitrogen, oxygen and sulfur heteroatom.Five-membered ring has zero to two two keys, and six have zero to three two keys with heptatomic ring.Term " heterocyclic radical " also comprises bicyclic radicals, and wherein heterocyclic ring and 4 to 7 yuan (preferred 4 to 6 yuan) fragrance or non-aromatic carbocycle or another monocyclic heterocycles base condense.Heterocyclic radical of the present disclosure can be connected with parent molecular moiety through carbon atom in the group or nitrogen-atoms.The example of heterocyclic radical is including, but not limited to benzothienyl, furyl, imidazole radicals, indolinyl, indyl; Isothiazolyl ， isoxazolyl, morpholinyl ， oxazolyl, piperazinyl; Piperidyl, pyrazolyl, pyridine radicals, pyrrolidinyl; Pyrrolopyridinyl, pyrrole radicals, thiazolyl, thienyl and tetrahydro-1,4-thiazine base.Heterocyclic radical of the present disclosure can be chosen wantonly by one, two, three, four or five and independently be selected from following substituent group replacement: thiazolinyl, alkoxyl, alkoxy carbonyl group, alkyl; Aryl, aralkyl, aryloxy group, cyanic acid; The cyanic acid alkyl, halogen, halogenated alkoxy, haloalkyl; Second heterocyclic radical, heterocyclic radical alkyl, nitro and oxo; Wherein the heterocyclic radical of the aryl moiety of aryl, aralkyl and aryloxy group, second heterocyclic radical and heterocyclic radical alkyl part can further choose by one, two, three, four or five and independently be selected from following substituent group replacement: thiazolinyl; Alkoxyl, alkyl, cyanic acid; Halogen; Halogenated alkoxy, haloalkyl, nitro and oxo.

The term " heterocyclic radical alkyl " that this paper uses is meant by one, two or three substituted alkyl of heterocyclic radical.

The term " heterocyclic radical carbonyl " that this paper uses is meant the heterocyclic radical that is connected with parent molecular moiety through carbonyl.

The term " heterocyclyloxy base " that this paper uses is meant the heterocyclic radical that is connected with parent molecular moiety through oxygen atom.

The term " heterocyclyloxy base carbonyl " that this paper uses is meant the heterocyclyloxy base that is connected with parent molecular moiety through carbonyl.

The term " hydroxyl " that this paper uses is meant-OH.

The term " nitro " that this paper uses is meant-NO ₂

Term " the NR that this paper uses ^aR ^b" be meant two radicals R that are connected with parent molecular moiety through nitrogen-atoms ^aAnd R ^bR ^aAnd R ^bBe independently selected from hydrogen, alkoxyl, alkyl, aryl, aralkyl, cycloalkyl, (cycloalkyl) alkyl, haloalkyl, heterocyclic radical and heterocyclic radical alkyl.

The term " (NR that this paper uses ^aR ^b) sulfonyl " and be meant through sulfonyl be connected with parent molecular moiety-NR ^aR ^b

Term " the NR that this paper uses ^eR ^f" be meant two radicals R that are connected with parent molecular moiety through nitrogen-atoms ^eAnd R ^fR ^eAnd R ^fBe independently selected from hydrogen, alkyl, aryl, aralkyl and heterocyclic radical; Wherein the aryl moiety of aryl, aralkyl and heterocyclic radical are optional is replaced by one or two substituent group that is independently selected from alkoxyl, alkyl and halogen.

The term " (NR that this paper uses ^eR ^f) carbonyl " and be meant through carbonyl be connected with parent molecular moiety-NR ^eR ^f

The term " oxo " that this paper uses is meant=O.

The term " sulfonyl " that this paper uses is meant SO ₂

Chemical compound of the present disclosure can exist with prodrug form.Term " prodrug " expression that this paper uses can be converted into the chemical compound of parent compound through hydrolysis in blood in the quick body.Prodrug of the present disclosure comprises the ester of the hydroxyl on the parent molecule, the ester of the carboxyl on the parent molecule and the amide of the amine on the parent molecule.

Chemical compound of the present disclosure can exist with pharmaceutical acceptable salt.The salt or the zwitterionic form of term " officinal salt " the expression disclosure chemical compound that this paper uses; It is that water or oil dissolve or dispersible; In reliable medical judgment scope, the tissue that it is suitable for contacting the patient does not have excessive toxicity, zest, allergy or other problem or complication; Match with rational benefit/dangerous ratio, and be effective for their intended purpose.Salt can prepare during the final separation of chemical compound and purification, or comes preparation separately through the reaction of suitable basic functionality and appropriate acid.Representational acid-addition salts comprises acetate, adipate, alginate, citrate, aspartate, benzoate, benzene sulfonate, disulfate, butyrate, camphorate, camsilate; Digluconate, glycerophosphate, Hemisulphate, enanthate, caproate, formates, fumarate; Hydrochlorate, hydrobromate, hydriodate, 2-isethionate, lactate, maleate, sym-toluenesulfonic acid salt; Mesylate, naphthylene sulfonate, nicotinate, 2-naphthalene sulfonate, oxalates, palmitate, pectinic acid salt; Persulfate, 3-phenylpropionic acid salt, picrate, pivalate, propionate, succinate, tartrate; Trichloroacetate, trifluoroacetate, phosphate, glutamate, Glu, bicarbonate, tosilate and hendecane hydrochlorate.The example that can be used to form the acid of pharmaceutically acceptable addition salts comprises for example for example oxalic acid, maleic acid, succinic acid and citric acid of hydrochloric acid, hydrobromic acid, sulphuric acid and phosphoric acid and organic acid of mineral acid.

Base addition salts can prepare during the final separation of chemical compound and purification, through acidic-group and appropriate base for example the carbonate or the bicarbonate of hydroxide, metal cation react, or with ammonia or organic primary, second month in a season or reactive tertiary amine.The cation of officinal salt comprises lithium, sodium, potassium, calcium, magnesium and aluminium cations, and atoxic quaternary ammonium cation, ammonium for example, tetramethylammonium, etamon, methylamine; DMA, trimethylamine, triethylamine, diethylamine, ethamine, TBuA; Pyridine, N, N-dimethylaniline, N-methyl piperidine, N-methylmorpholine, dicyclohexylamine; Procaine, dibenzylamine, N, N-dibenzyl phenethylamine and N, N '-dibenzyl-ethylenediamin.Other the representational organic amine that can be used for forming base addition salts comprises ethylenediamine, ethanolamine, diethanolamine, piperidines and piperazine.

The term " anti-HCV activity " that this paper uses is meant that chemical compound can effectively treat HCV virus.

Term " chemical compound of the present disclosure " and equivalents are meant the chemical compound and its pharmaceutically acceptable enantiomer, diastereomer and salt of formula (I).Equally, for intermediate, its implication comprises their salt under the situation of in context, allowing.

Term " patient " comprises people and other mammal.

Term " pharmaceutical composition " is meant a kind of compositions, and it comprises the combination of chemical compound of the present disclosure and at least a other medicines carrier, and carrier is auxiliary agent, excipient or carrier, diluent for example, antiseptic; Filler, flowing regulator, disintegrating agent, wetting agent, emulsifying agent; Suspending agent, sweeting agent, flavoring agent, spice; Antibacterial, antifungal, lubricant and partitioning agent, this depends on mode of administration and dosage form.Can use and for example list in Remington ' s Pharmaceutical Sciences, 18th ed., Mack Publishing Company, Easton, the component among the PA (1999).

The phrase " pharmacy is acceptable " that this paper uses refers to those chemical compounds, material, compositions and/or dosage form; In reliable medical judgment scope; It is fit to contact with patient's tissue; Do not have undue toxicity, zest, allergy or other problem or complication, match with rational danger/benefit ratio.

Term " treatment effective dose " is meant is enough to show the for example total amount of each active component of virus load continuous decrease of significant patient's benefit.When being applied to the single-activity component separately, this term is meant independent component.In the time of in being applied to combination, this term is meant the number of combinations of the active component that can cause therapeutic effect, no matter combination gives, gives continuously or gives simultaneously.

Term " treatment " is meant: (i) ward off disease, obstacle or disease occur in the patient, it possibly suffered from this disease, obstacle and/or disease by indication, but does not also make a definite diagnosis; (ii) suppress disease, obstacle or disease, i.e. its development of retardation; And/or (iii) palliate a disease, obstacle or disease, promptly cause the decline of disease, obstacle and/or disease.

Be used to name under the situation of disclosure chemical compound symbol P1 ', P1, P2, P2 that this paper uses ^*, P3 and the bonded amino acid residue of P4 labelled protein enzyme inhibitor relative position (with respect to the binding site of native peptides cracking substrate).Cracking occurs between the P1 and P1 ' of natural substrates, and wherein non-main positions is represented from the C-of the natural cracking site of peptide terminal initial to the terminal aminoacid that extends of N-; And the N-end that main positions identifies from cracking site sends the terminal extension to C-.For example, P1 ' is meant first position (being terminal first position of N-) of the right hand end of the C-end that leaves cracking site; And P1 is from the left side open numbering of the terminal cracking site of C-, and P2 is from terminal second the initial position of C-, or the like).(referring to Berger A.& Schechter I., Transactions ofthe Royal Society London series (1970), B257, 249-264].

Asymmetric center is present in the chemical compound of the present disclosure.For example, chemical compound can comprise the P1 cyclopropyl unit of following formula:

C wherein ₁And C ₂The asymmetric carbon atom of representing 1 and 2 positions of cyclopropyl rings separately.

(1R，2S) (1S，2R)

R ²With carbonyl homonymy R ²With the carbonyl homonymy

(1R，2R) (1S，2S)

R ²With amide groups homonymy R ²With the amide groups homonymy

Should be appreciated that the disclosure comprises having all stereochemical forms or its mixture that suppresses HCV protease ability.

Can also there be different stable structure forms in some chemical compound of the present disclosure, and this form is can be isolating.Reversing unsymmetry is because effect is revolved in the resistance around the asymmetric singly-bound, for example because sterically hindered or ring strain can separate different conformers.The disclosure comprises each conformer and its mixture of these chemical compounds.

Can there be zwitterionic form in some chemical compound of the present disclosure, and the disclosure comprises every kind of zwitterionic form and its mixture of these chemical compounds.

When being used to treat, formula (I) chemical compound that can treat effective dose with original chemicals form with and officinal salt, this can provide the active component of pharmaceutical composition.Correspondingly, the disclosure further provides pharmaceutical composition, and it comprises formula (I) compound or pharmaceutically acceptable salt thereof and one or more pharmaceutically suitable carrier, diluent or excipient of treating effective dose.The chemical compound of formula (I) and its officinal salt are as stated.Carrier, diluent or excipient must be acceptable, with other component of preparation contradiction not, and harmless to its receiver.According to another aspect of the present disclosure, the method for useful in preparing drug formulations is provided, this method comprises: with compound or pharmaceutically acceptable salt thereof and one or more pharmaceutically suitable carrier, diluent or the mixed with excipients of formula (I).

Pharmaceutical preparation can provide with unit dosage form, and per unit dosage contains the active component of quantification in advance.Disease for prevention and treatment HCV mediation; Typical single current system method is: give every day about 0.01 with about 250 milligrams every kilogram (" mg/kg ") body weight between the disclosure chemical compound of dosage level, preferably between every day about 0.05 and about 100mg/kg body weight.Typically, pharmaceutical composition of the present disclosure can give about 1 to about 5 times every day, perhaps transfusion continuously.This form of medication can be used as chronic or acute treatment.Can make up the amount of active ingredients for preparing single dosage form with carrier mass can change, and this depends on the order of severity, administration time, the route of administration of the disease of treating, disease, drainage rate, treatment persistent period and patient's age, sex, weight and the disease of the chemical compound that uses.Preferred unit dose formulations is to contain the daily dose of the above-named active component of this paper or those preparations of sub-doses or its suitable part.Usually, with coming begin treatment less than the low dose of the optimal dose of chemical compound basically.Then, increase dosage slightly, till reaching optimum efficiency in this case.Usually, most preferably, do not cause that to obtain the effective result of antiviral usually any concentration level bad or harmful side effect gives chemical compound.

When compositions of the present disclosure comprises the combination of disclosure chemical compound and one or more other treatment or preventative medicament; Chemical compound and other medicament usually with in the single current system method scheme usually the dosage level between about 10 to 150% of dosage exist, and more preferably between about 10 and 80%.

Pharmaceutical preparation can be suitable for any suitable pathways administration, for example oral (comprising oral cavity or Sublingual), rectum, nose, part (comprising buccal, Sublingual or transdermal), vagina or parenteral (comprise in subcutaneous, Intradermal, intramuscular injection, intraarticular, the synovial membrane, in the breastbone, in the sheath, intralesional, intravenous or intradermal injection or transfusion) approach.This preparation can for example make active component and carrier or excipient composition through the known any method preparation of pharmaceutical field.

Being suitable for oral pharmaceutical preparation can provide with the discrete unit form, for example capsule or tablet; Powder or granule; Solution in water or on-aqueous liquid or suspension; Edible foam or whips; Or oil-in-water liq latex or water-in-oil emulsion.

For example, for the oral administration of tablet or capsule form, can for example ethanol, glycerin, water or the like combine with oral, nontoxic pharmaceutical acceptable inert carriers with the active medicine component.Powder is prepared as follows: chemical compound is milled to the powder of suitable size, with similar ground pharmaceutical carriers for example edible carbohydrate for example starch or mannitol mix.Can also there be flavoring agent, antiseptic, dispersion and coloring agent.

Capsule can be prepared as follows: prepare aforesaid mixture of powders, and be filled in the gel shell of shaping.Can with fluidizer and lubricant for example colloidal silica, Pulvis Talci, magnesium stearate, calcium stearate or solid polyethylene glycol join in the mixture of powders, then fill.When ingestible capsule, can also add disintegrate or solubilizing agent for example agar, calcium carbonate or sodium carbonate, to improve the utilization rate of medicine.

In addition, when requiring or need, can also suitable junction mixture, lubricant, disintegrating agent and colorant combination be advanced in the mixture.Suitable bonding agent comprises starch, gel, and natural sugar is glucose or beta lactose for example, and corn sweetener, natural and paragutta be arabic gum, tragacanth or sodium alginate for example, carmellose, Polyethylene Glycol or the like.The lubricant that is used for these dosage forms comprises enuatrol, sodium chloride or the like.Disintegrating agent includes but not limited to starch, methylcellulose, agar, bentonite, xanthan gum or the like.Following preparation tablet: for example prepare mixture of powders, pelletize or process rod add lubricant and disintegrating agent, and extruding becomes tablet.Be prepared as follows mixture of powders: the chemical compound of suitable pulverizing is mixed with aforesaid diluent or substrate; Optional mix with following: bonding agent is carmellose, alginic acid, gel or polyvinylpyrrolidone for example, and the solution blocker is paraffin hydrocarbon, absorption enhancer for example bentonite, Kaolin or calcium hydrogen phosphate of quaternary salt and/or absorbent for example more for example.Can mixture of powders be carried out pelletize as follows:, and force it to pass through sieve with the bonding agent solution wetted of syrup, gelatinized corn starch, acadia rubber cement or cellulose or polymeric material for example.As the alternative method of pelletize, can make mixture of powders flow through tablet machine, the result is that the rod that not exclusively is shaped is split into granule.By means of adding stearic acid, stearate, Pulvis Talci or mineral oil, can granule is lubricated, to prevent to cling the tablet loose tool.Then lubricated mixture is compressed into tablet.Can also chemical compound of the present disclosure be combined with runny inert carrier, and directly be compressed into tablet, need not experience pelletize or form the rod step.Can provide cleaning or opaque protective coating (the polishing coating by sheet glue-line, sugared coating or the polymerization material coating and the paraffin of sealing is formed).Can dyestuff be joined in these coatings, to distinguish different UDs.

The liquid oral that can prepare dosage unit form is solution, syrup and elixir for example, so that comprise the chemical compound of predetermined quantity to determined number.Can be through compound dissolution be prepared syrup in the aqueous solution of suitable seasoning, and elixir is through using nontoxic excipient to prepare.Can also add solubilizing agent and emulsifying agent (the for example pure and mild polyoxyethylene sorbitol ether of ethoxylation isooctadecane), antiseptic, flavouring additive (for example Oleum menthae or natural sweetener or glucide or other artificial sweetening), or the like.

If suitable, the oral dosage units preparation can be micro-encapsulated.Can also prepare prolongation or sustained release formulation, for example, through in polymer, paraffin or the like, being coated with stain or embedding particulate matter.

Can also be with the chemical compound and its officinal salt of the form giving construction (I) of liposome delivery system, for example small unilamellar vesicle, big unilamellar liposome and MLV.Liposome can for example cholesterol, octadecane amine stearylamine or phosphatidylcholine form by various phospholipid.

The chemical compound of formula (I) also can utilize the monoclonal antibody of the bonded single carrier format of compound molecule to send with its officinal salt.This chemical compound also can combine with the soluble polymer as the drug target carrier.This polymer can comprise polyvinylpyrrolidone, pyran co-polymer, gather hydroxypropyl methacryl amide phenol, gather hydroxyethyl agedoite phenol or by the substituted PEO polylysine of palmityl residue.In addition; This chemical compound can combine with biodegradable polymer; Be used to realize that controlled delivery of pharmaceutical agents discharges, for example polylactic acid, poly epsilon caprolactone lactone (polepsilon caprolactone), poly butyric, poe, gather acetal, gather dihydropyran, the block copolymer of polybutylcyanoacrylate and crosslinked or amphipathic hydrogel.

Can the pharmaceutical preparation that be suitable for transdermal administration be provided with discrete patch form (predetermined a period of time that keeps contacting closely prolongation) with receiver's epidermis.For example, utilize usually at Pharmaceutical Research, 3 (6), 318 (1986) the middle ionotherapies of describing, active component can be sent from paster.

Can the pharmaceutical preparation that be suitable for topical be formulated as ointment, emulsifiable paste, suspension, lotion, powder, solution, ointment, gel, spray, aerosol or oil.

For treatment eyes or other outside organization, for example oral cavity and skin are preferred, with the administered preparation of local ointment or emulsifiable paste.When the preparation ointment, active component can be used with the ointment base that paraffin or water can dissolve each other.Perhaps, can active component be formulated as emulsifiable paste with oil-in-water type cream base or water-in-oil type substrate.

The pharmaceutical preparation that is suitable for the topical administration eyes comprises eye drop, wherein solubilization of active ingredient or be suspended in the suitable carrier, especially aqueous solvent.

The pharmaceutical preparation that is suitable for the topical administration oral cavity comprises lozenge, pastille and collutory.

Being suitable for rectum administered agents preparation can provide with suppository or enema forms.

The pharmaceutical preparation (wherein carrier is a solid) that is suitable for nasal administration comprises having the for example coarse powder of 20 to 500 micrometer range particle diameters, and it is to adopt the mode of snuffing to give, promptly from passing through nasal cavity near sucking fast the powder container of nose.Spray into or the appropriate formulation (wherein carrier is a liquid) of nasal drop form for nose, comprise the water or the oil solution of active component.

The pharmaceutical preparation that is suitable for inhalation comprises particulate or mist agent, and it can utilize aerosol apparatus, nebulizer or the insufflator gageable, the dosage pressurization of various types to produce.

The pharmaceutical preparation that is suitable for vagina administration can provide with the form of vaginal suppository, plug, emulsifiable paste, gel, ointment, foam or spray.

The pharmaceutical preparation that is suitable for parenteral comprises moisture and anhydrous aseptic parenteral solution, and it can contain antioxidant, buffer, antibacterial and solute, makes preparation and intended recipient's blood isoosmotic pressure; With the aseptic suspension of moisture and non-water that can comprise suspending agent and thickening agent.Preparation can also be provided in UD or the multi-dose container, the ampoule and the phial that for example seal, and can be kept under lyophilization (lyophilizing) condition, before using, only need to add aseptic liquid-carrier, for example water is used for injecting at once.Interim injection and suspension can be used sterilized powder, granule and preparation tablets.

Should be appreciated that except the top component of mentioning especially, preparation can comprise this area for other habitual medicament of said preparation type, for example is suitable for those oral preparations and can comprises flavoring agent.

Below table 1 listed can be with some illustrative example of disclosure chemical compound administered compound.In combined therapy, chemical compound of the present disclosure can give with other anti-HCV activity chemical compound, can unite or give separately, or give through chemical compound is combined as compositions.

Table 1

Trade name	Physiology's classification	The type of inhibitor or targeting	Source company
				NIM811	?	The cyclophilin inhibitor	Novartis
Zadaxin (Zadaxin)	?	Immunomodulator	Sciclone
				Suvus	?	Methylene blue	Bioenvision
Actilon(CPG10101)	?	The TLR9 agonist	Coley
				Batabulin(T67)	Anticarcinogen	The 'beta '-tubulin inhibitor	Tularik?Inc.，South?San Francisco，CA
ISIS?14803	Antiviral	Antisense	ISIS?Pharmaceuticals?Inc， Carlsbad，CA/Elan Phamaceuticals?Inc.，New York，NY
				Summetrel	Antiviral	Antiviral	Endo?Pharmaceuticals Holdings Inc.，Chadds Ford，PA
GS-9132(ACH-806)	Antiviral	The HCV inhibitor	Achillion/Gilead
				Pyrazolopyrimidine compound and salt are obtained from WO-2005047288 on May 26th, 2005	Antiviral	The HCV inhibitor	Arrow?Therapeutics?Ltd.
Levovirin (Levovirin)	Antiviral	The IMPDH inhibitor	Ribapharm?Inc.，Costa Mesa，CA
				Merimepodib (VX-497)	Antiviral	The IMPDH inhibitor	Vertex?Pharmaceuticals Inc.，Cambridge，MA
XTL-6865(XTL-002)	Antiviral	Monoclonal antibody	XTL?Biopharmaceuticals Ltd.，R ^ehovot，Isreal
				Telaprevir (VX-950，LY-570310)	Antiviral	The NS3 serpin	Vertex?Pharmaceuticals Inc.，Cambridge，MA/Eli Lilly?and?Co.Inc.， Indianapolis，IN

Trade name	Physiology's classification	The type of inhibitor or targeting	Source company
				?HCV-796	Antiviral	NS5B replicative enzyme inhibitor	Wyeth/Viropharma
?NM-283	Antiviral	NS5B replicative enzyme inhibitor	Idenix/Novartis
				?GL-59728	Antiviral	NS5B replicative enzyme inhibitor	Gene?Labs/Novartis
?GL-60667	Antiviral	NS5B replicative enzyme inhibitor	Gene?Labs/Novartis
				?2’C?MeA	Antiviral	NS5B replicative enzyme inhibitor	Gilead
?PSI?6130	Antiviral	NS5B replicative enzyme inhibitor	Roche
				?R1626	Antiviral	NS5B replicative enzyme inhibitor	Roche
The acid of 2 ' C methyladenosine	Antiviral	NS5B replicative enzyme inhibitor	Merck
				?JTK-003	Antiviral	The RdRp inhibitor	Japan?Tobacco?Inc.， Tokyo，Japan
Levovirin (Levovirin)	Antiviral	Ribavirin	ICN?Pharmaceuticals， Costa?Mesa，CA
				Ribavirin	Antiviral	Ribavirin	Schering-Plough Corporation，Kenilworth， NJ
Ribavirin precursor (Viramidine)	Antiviral	The ribavirin prodrug	Ribapharm?Inc.，Costa Mesa，CA
				?Heptazyme	Antiviral	Ribozyme	Ribozyme Pharmaceuticals Inc， Boulder，CO
?BILN-2061	Antiviral	Serpin	Boehringer Ingelheim Pharma KG，Ingelheim， Germany
				?SCH?503034	Antiviral	Serpin	Schering?Plough
?Zadazim	Immunomodulator	Immunomodulator	SciClone?Pharmaceuticals Inc.，San?Mateo，CA
				?Ceplene	Immunomodulator	Immunomodulator	Maxim?Pharmaceuticals Inc.，San?Diego，CA
?CellCept	Immunosuppressant	HCV IgG immunosuppressant	F..Hoffmann-La Roche LTD，Basel，Switzerland

Trade name	Physiology's classification	The type of inhibitor or targeting	Source company
				Civacir	Immunosuppressant	HCV IgG immunosuppressant	Nabi?Biopharmaceuticals Inc.，Boca?R ^aton，FL
Albuferon-α	Interferon	Albumin IFN-α 2b	Human?Genome?Sciences Inc.，Rockville，MD
				Infergen?A	Interferon	IFN?alfacon-1	InterMune Pharmaceuticals?Inc.， Brisbane，CA
Omega?IFN	Interferon	IFN-ω	Intarcia?Therapeutics
				IFN-β and EMZ701	Interferon	IFN-β and EMZ701	Transition?Therapeutics Inc.，Ontario，Canada
Rebif	Interferon	IFN-β1a	Serono，Geneva， Switzerland
				Roferon?A	Interferon	IFN-α2a	F..Hoffmann-La Roche LTD，Basel，Switzerland
Intron?A	Interferon	IFN-α2b	Schering-Plough Corporation，Kenilworth， NJ
				Intron A and Zadaxin (Zadaxin)	Interferon	IFN-α 2b/ α 1-thymosin	R ^egeneRx Biopharmiceuticals Inc.， Bethesda，MD/ SciClone Pharmaceuticals Inc，SanMateo，CA
Rebetron	Interferon	IFN-α 2b/ ribavirin	Schering-Plough Corporation，Kenilworth， NJ
				Actimmune	Interferon	INF-γ	InterMune?Inc.，Brisbane， CA
Interferon-beta	Interferon	Interferon-beta-1a	Serono
				Multiferon	Interferon	Long-acting IFN	Viragen/Valentis

Trade name	Physiology's classification	The type of inhibitor or targeting	Source company
				Wellferon	Interferon	Lymphoblastoid IFN-α n1	Glaxo?SmithKline?plc， Uxbridge，UK
Omniferon	Interferon	Natural IFN-α	Viragen?Inc.，Plantation， FL
				Pai Luoxin	Interferon	The IFN-α 2a of PEGization	F..Hoffmann-La Roche LTD，Basel，Switzerland
Pai Luoxin and Ceplene	Interferon	The IFN-α 2a/ immunomodulator of PEGization	Maxim?Pharmaceuticals Inc.，San?Diego，CA
				Pai Luoxin and ribavirin	Interferon	The IFN-α 2a/ ribavirin of PEGization	F..Hoffmann-La Roche LTD，Basel，Switzerland
PEG-Intron	Interferon	The IFN-α 2b of PEGization	Schering-Plough Corporation，Kenilworth， NJ
				The PEG-Intron/ ribavirin	Interferon	The IFN-α 2b/ ribavirin of PEGization	Schering-Plough Corporation，Kenilworth， NJ
IP-501	The liver protection	Fibrosis	Indevus?Pharmaceuticals Inc.，Lexington，MA
				IDN-6556	The liver protection	The caspase inhibitor	Idun?Pharmaceuticals Inc.，San?Diego，CA
ITMN-191(R-7227)	Antiviral	Serpin	InterMune Pharmaceuticals?Inc.， Brisbane，CA
				GL-59728	Antiviral	NS5B replicative enzyme inhibitor	Genelabs
ANA-971	Antiviral	The TLR-7 agonist	Anadys
				TMC-465350	Antiviral	Serpin	Medivir/ Tibotec

Chemical compound of the present disclosure can also be used as laboratory reagent.For the design of virus replication test, the checking and the structure biology research of animal experiment system, this chemical compound can help to provide research tool, thereby increases the understanding to the HCV pathogenic mechanism.Further, chemical compound of the present disclosure can be effective to set up or confirm the binding site of other antiviral compound, for example, utilizes competitive inhibition.

Chemical compound of the present disclosure can also be used to treat or prevent the viral pollution of material; Therefore can reduce the laboratory that touches this material or medical worker or patient's risk from viral infection, this material is blood, tissue, operating theater instruments and clothing, experimental apparatus and clothing and blood collecting or infusion set and material for example.

When through synthetic method or through metabolic process (comprising those (in the bodies) that are present in the human or animal body or the process of external existence) preparation, the disclosure comprises the have formula chemical compound of (I).

Be generally used for differentiating that the chemical abbreviations of chemical compound disclosed herein comprises: Bn: benzyl; Boc: tertbutyloxycarbonyl { Me ₃COC (O) }; BSA: bovine serum albumin; CDI: N,N'-carbonyldiimidazole; DBU:1,8-diazabicyclo [5.4.0]-11-7-alkene; CH ₂Cl ₂=DCM: dichloromethane; TBME: t-butyl methyl ether; DEAD: diethylazodicarboxylate; DIAD: diisopropyl azo group dicarboxylic ester; DIEA: diisopropylethylamine; DIPEA: diisopropylethylamine; The 4-DMAP:4-dimethyl aminopyridine; DCC:1, the 3-dicyclohexylcarbodiimide; DMF: dimethyl formamide; DMSO: dimethyl sulfoxine; DPPA: diphenylphosphine acyl azide; Et: ethyl; EtOH: ethanol; EtOAc: ethyl acetate; Et ₂O: diethyl ether; Grubb ' s catalyst: two (tricyclohexyl phosphine) benzal chloride ruthenium (IV); The secondary catalyst of Grubb ' s: tricyclohexyl phosphine [1, two (2,4, the 6-trimethylphenyl)-4 of 3-, 5-glyoxalidine-2-subunit] [benzal] ruthenous chloride (IV); HATU: [O-(7-azepine benzo triazol-1-yl)-N, N, N ', N '-tetramethylurea hexafluorophosphate; HBTU: [O-(1H-BTA-1-yl)-N, N, N ', N '-tetramethylurea hexafluorophosphate; HOBT, I-hydroxybenzotriazole; HOAT:1-hydroxyl-7-azepine BTA; HPLC: HPLC; MS: mass spectrum; Me: methyl; MeOH: methanol; The NMM:N-methyl morpholine; The NMP:N-crassitude; Pr: propyl group; PPA: polyphosphoric acid; TBAF: four-normal-butyl ammonium fluoride; 1,2-DCE or DCE:1,2-dichloroethanes; TFA: trifluoroacetic acid; THF: oxolane.

The initiation material that is used for synthetic disclosure chemical compound known by those skilled in the art, and can easily prepare or commercially available.

The following method of listing below is that the illustrative purpose provides, and is not the scope that is used for limiting claim.Should be realized that, when this chemical compound of preparation, must use habitual protection base to protect functional group, remove the protection base then, chemical compound of the present disclosure is provided.Relevantly known by those skilled in the art according to the details of protection base of using of the present disclosure.

Shown in reaction scheme 1, can use for example dipeptides (1) of intermediate of the present disclosure, the chemical compound of preparation formula (I).In the first step of this method, use for example HCl of acid, at solvent for example in the ether, the nitrogen deprotection that Boc that will (1) protects provides corresponding unhindered amina 2.Use for example HATU of coupling agent subsequently, at solvent for example in the dichloromethane, amine (2) can with aminoacid (3) coupling, tripeptide intermediate (4) is provided.It should be noted that in some cases intermediate for example (3) is commercially available, perhaps this chemical compound can be easily through methods known in the art, prepare with raceme or chirality mode.In the structure of formula (I) chemical compound, the crucial conversion is big cyclization process, and wherein the intermediate of universal architecture (4) changes the intermediate of universal architecture (5) into.In the conventional embodiment of quoting, intermediate (4) is converted into the influence that (5) possibly receive intramolecular olefin metathesis reactions.This kind be reflected at this area by abundant establishment, therefore, developed many olefin metathesis catalysts, and commercially available.For example, diene (4) is converted into macro ring (5) and possibly receives influence with the processing of sufficient amount Grubb ' s first generation olefin metathesis catalyst (at solvent for example in dichloromethane or the dichloroethanes).In some example of the conversion of (4) to (5), possibly need the reacting by heating mixture, so that realize this cyclization process.Then, through two-stage process, the chemical compound that intermediate (5) changes formula (I) into is (7) for example.In the first step of this method, the ester functional group of intermediate (5) is hydrolyzed to corresponding carboxylic acid (6).This conversion can realize through saponification, wherein in the mixture of THF, first alcohol and water with alkali for example Lithium hydrate handle (5).Through shown in the simple coupling reaction of sulfonamide, the acid that obtains (6) can change the chemical compound of formula (I) into.For example, this area is used for a long time to be, for example in the dichloromethane, handles carboxylic acid for example (6) with CDI at solvent, and original position produces reactive intermediate, when handling this reactive intermediate with sulfonamide, (7) is provided, i.e. the chemical compound of formula (I).

Reaction scheme 1

If in above-mentioned last coupling process promptly in (6) to the conversion of (7), R ⁷SO ₂NH ₂Body is a sulfamide derivative, for example R ^aR ^bNSO ₂NH ₂, can use other couling process so.Therein, use the for example two hexamethyldisilane lithiums of alkali (at solvent for example among the THF), at first deprotonation of sulfonamide intermediate (1) (reaction scheme 2).THF solution with the sulfonamide anions that obtains (2) joins in the above-mentioned reactant mixture that contains the activating carboxy acid then.Then this reactant mixture was stirred several hours, the acyl group sulfonamide (7) that needs is provided.

Reaction scheme 2

The chemical compound of formula (I) can also change other formula I chemical compound into, such as this paper description.The example of this method is shown in the reaction scheme 3, and the chemical compound (1) (it carries the Boc group in the P4 position) of its Chinese style I changes the chemical compound (3) (wherein said chemical compound carries urea groups in the P4 position) of formula I into.(1) can carry out with two-stage process to the conversion of (3), at first, at solvent for example in the dichloromethane, with acid for example TFA handle (1), (1) is converted into amine (2).In the presence of monovalent alkali, can handle the amine tfa salt that obtains with isocyanates, the chemical compound (3) of formula I is provided, wherein P3 partly uses the urea end-blocking.Like previous institute note, one skilled in the art will recognize that intermediate (2) can be as the initiation material of preparation formula (I) chemical compound, wherein the P3 group is with amide or carbamate end-blocking.Use forms the standard conditions of said P4 functional group by amine, can obtain the structure of said formula (I) chemical compound.

Reaction scheme 3

The non-limiting method of preparation P2 intermediate and formula (I) chemical compound is shown in the following reaction scheme.Said intermediate, reaction condition and the method that in specific embodiment, provides is applicable to the chemical compound of other substitution pattern widely.For example, the P2 that in formula (I) chemical compound of reaction scheme 4, finds is unitary synthetic, can prepare according to defined synthesis path.Therein, with organometallic reagent for example RMgBr (perhaps alkyl or aryl lithium species, perhaps alkyl or aryl zinc species) handle commercially available N-Boc-4-oxo-L-proline, intermediate (2) is provided, wherein R is carried in the C4 position of proline ³Substituent group and free three grades of hydroxyls.Intermediate (2) changes the chemical compound of formula (I) into then.

Reaction scheme 4

The chemical compound of formula (I) (R wherein ⁴Be hydrogen) can be by corresponding formula (I) chemical compound (R wherein ⁴Be hydroxyl) synthetic.Perhaps, the chemical compound of formula (I) (R wherein ⁴Be hydrogen) can be by being used for preparation formula (I) chemical compound (R wherein ⁴Be hydroxyl) any intermediate synthetic.For example, R wherein ⁴Formula (I) chemical compound that is hydrogen can prepare shown in reaction scheme 5.Said chemical compound (3) can be prepared by corresponding hydroxy analogs (1).By chemical compound for example (1) form chemical compound for example the method for (3) require the reduction or the deoxidation of proline C4 hydroxyl.For the reduction and/or the deoxidation, the particularly tertiary alcohol of alcohol, have considerable definite technology can provide by chemical compound for example (1) form chemical compound for example (3).For example, in following list of references, reported the pure corresponding alkane (method 1 of reaction scheme 5) that directly is reduced to: J.Org.Chem.2001,66,7741.Described alcohol therein and be reduced to alkane, wherein for example handled said alcohol with the Indium-111 chloride of chloro diphenyl silane and catalytic quantity in the dichloroethanes, corresponding alkane is provided at solvent.Said reaction can at room temperature be carried out, but possibly require heating in some cases.By formula (I) chemical compound (R wherein ⁴Be hydroxyl) form formula (I) chemical compound (R wherein ⁴Be hydrogen) other method such as 2 demonstrations of method of reaction scheme 5.Therein, chemical compound for example (1) at first change activation alcohol (2) into and these intermediate are reduced to corresponding alkane (for example using the Barton method of deoxidation).Perhaps, the activation alcohol shown in (2) can be reduced into corresponding alkane, for example uses Reducing agent.These methods of the conversion of (1) in reaction scheme 5 to (3) are well-known to those skilled in the art.

Reaction scheme 5

It should be noted that it is fully to establish in this area that organometallic reagent partly carries out addition (reaction scheme 6) to the ketone of proline derivative 1.For example, Hruby and colleague (J.Org.Chem.2001,66,3593) have described phenyl-magnesium-bromide the intermediate of the universal architecture (1) of reaction scheme 6 have been carried out addition.These discoveries provide following evidence: when using tert-butyl group ester group as C ₂During the protection base of carboxy moiety, obtain the optimum yields of needed 1,2 addition product ((2), reaction scheme 6).In addition, in the X-ray crystal diffraction form about the spatial chemistry result of additive reaction, this work provides clear evidence.Specifically, because above-mentioned RMgBr to ketone (1) addition, obtains single product, wherein C4 hydroxyl and C2 carboxyl are assumed to five-membered ring cis relative orientation on every side.Structure determination deducibility thus, under the condition of the structure (1) of reaction scheme 6, R ³M is α to the surface selectivity that the ketone of (1) carries out addition.That is, organic metal is to reverse side (bottom surface) addition selectively of carbonyl in (1), can provide have show the stereochemical corresponding tertiary alcohol (2).

Reaction scheme 6

The above-mentioned work specification of Hruby the addition (reaction scheme 6) of concrete RMgBr to the derivant of (1).Yet various RMgBrs are included in the disclosure the addition of proline (1).In this area; Describing organometallic reagent (comprising RMgBr) is considerable to the document that ketone carries out addition; And be summarised in the summary, for example: Comprehensive Organic FunctionalGroup Transformations.2 volume: people such as Synthesis:Carbon with one heteroatomattached by a single bond.Alan.R.Katritzky chief editor, nineteen ninety-five; 2.02 chapter, 37 pages.The reaction of this kind also is described among the Comprehensive Organic Synthesis.Barry M Trost chief editor, 1 volume: Additions to C-X pi-bonds (1 part), 1991.

The up-to-date research in this area provides further optimization RMgBr to the condition in the additive reaction of ketone, and these work can be used in the disclosure.For example, Ishihara and colleague (Org.Lett.2005,7 volumes, No.4,573) have described the formation and the application of magnesium salt Ate type complex recently.Magnesium salt Ate type complex R ³MgLi stems from RMgBr and lithium alkylide.Such as Ishihara description, in the reaction to ketone, these complex provide the outstanding productive rate of 1,2 addition product.At one separately in the research, Knochel and colleague (Angew.Chem Int.Ed.2006,45,497) described with the combining of organomagnesium reagent in use solubility group of the lanthanides salt LnCl for example ₃The existence of these group of the lanthanides salt can cause improving for the efficient of 1,2 additive reaction of carbonyl compound.In the simple addition to carbonyl compound, these work and the list of references of quoting have therein formed and have optimized the prior art of grignard reaction, and in the disclosure, serve as important source of data.

It should be noted that also a large amount of organo-metallic compounds can participate in the additive reaction to ketone.Being included in this work is for example aryl lithium, lithium alkylide and heteroaryl lithium reagent of reagent, and it is well-known in that carbonyl moiety is carried out 1,2 mode addition.For example, in Dondoni and colleague's nearest research (J.Org.Chem.2005,70,9257), use BuLi with the benzothiazole lithiumation, and with the C that obtains ₂-lithium species carry out addition with 1,2 mode to lactone.Utilize the benzothiazole of similar lithiumation, can expect the ketone (1) of reaction scheme 6 to be carried out addition, intermediate for example (2a) is provided with 1,2 mode.

One skilled in the art will recognize that derived from the organo-metallic compound of heterocycle Li such as oxazole and thiazole and imidazoles and also can participate in 1,2 additive reaction ketone (1).Have considerable document to define, and this information those skilled in the art can easily obtain for each the employed unique conditions in these heterocyclic ring systems.For example, in additive reaction, use organometallic reagent, require to use the lithium magnesium salt derived from benzoxazole Huo oxazole to ketone.The particular content of these current research of Bayh and colleague is described in J.Org.Chem., in 2005,70,5190.Benzoxazole can provide intermediate for example (2b) to the addition of the ketone (1) of reaction scheme 6.

There is lot of documents to describe to use and all kinds ofly ketone carried out addition derived from heterocyclic organometallic reagent.For example, the formation and its addition to simple lactone of lithiumation benzimidazole has been described in Behinda and colleague's work (Tet.Lett.42,2001,647).Equally, in additive reaction, use the benzimidazole of this lithiumation that intermediate for example (2c) can be provided to the ketone (1) of reaction scheme 6.In addition, the formation and they additive reaction for activating terephthalamide amine of the heteroaromatic compound of a series of lithiumations has been described in Kawasaki and colleague's nearest research (Bioorganic and Medicinal Chem.Lett.13,2003,87).Equally, in additive reaction, use the hetero-aromatic ring intermediate of these lithiumations that intermediate (2d-2k) can be provided to the ketone (1) of reaction scheme 6.

In 1,2 additive reaction, use also relevant with the disclosure derived from the organo-metallic compound of biaryl or heteroaryl-aryl system to ketone (1).The organometallic reagent of this kind can provide intermediate for example (21) and (2m) to the addition of ketone (1).It should be noted that in example of the present disclosure, possibly need synthetic biaryl or heteroaryl organo-metallic compound, in additive reaction, use subsequently the ketone (1) of reaction scheme 6.Those skilled in the art can understand the lot of documents of describing such organo-metallic compound and its precursor preparation.For example, Chinchilla and colleague's up-to-date summary (Chem.Rev.2004,104,2667) has been described metallize heterocyclic preparation and their application.The Essential Chemistry of preparation biaryl or heteroaryl-aryl system is usually used Suzuki class coupling reaction.A series of document descriptions that Gregory Fu delivers the state of the art of this coupling reaction, the further list of references of these lists of references is following: JACS 2004,126,1340; JACS, 2002,124,13662; Angew.Chem.Int.Ed.2002,41, No.11,1945; Angew.Chem.Int.Ed.2002,41, No.20,3910; JACS 2002,122, and 4020; JACS2001,123,10099; Org.Lett.2001, Vol.3, No.26,4295; Angew.Chem.Int.Ed.1998,37, No.24,3387.Except these a series of works, the key summary in this field is to obtain easily, and for example Rossi is at Synthesis 2004, No.15, the summary in 2419.

Embodiment

Now combine to describe the disclosure with some embodiments, these embodiments do not limit the scope of the present disclosure.On the contrary, the disclosure comprises all replacements, improvement and the equivalent that can be included in claims scope.Thus; The following example that comprises specific embodiments can illustrate a practice of the present disclosure; Certainly, these embodiment are purposes that some embodiment is described for example, and the explanation of the most useful and easy understanding of its process and notion aspect is provided.

Solution percent is expressed weight and volume relationship, and solution proportion is expressed volume and volume relationship, except as otherwise noted.On Bruker 300,400 or 500MHz spectrometer, write down nuclear magnetic resonance, NMR (NMR) spectrum; With the chemical shift (δ) of 1,000,000/report.According to Still ' s flash chromatography technology (J.Org.Chem.1978,43,2923), at silica gel (SiO ₂) on carry out flash chromatography.

Embodiment 1

The preparation of sulfonamide

In the THF of chloro sulfonyl isocyanate (1 equivalent) cold (20 ℃) agitating solution, add entry (1 equivalent) (in THF), and the solution that obtains is warming up to 0 ℃, preparation intermediate sulfamoyl chlorine.In this solution, add anhydrous Et ₃N (1 equivalent) then adds necessary secondary amine (1 equivalent).Reactant mixture is heated to room temperature, filters then, rotary evaporation filtrating obtains needed sulfonamide.Make said sulfonamide and carboxylic acid coupling then, needed acyl group sulfonamide is provided.

Embodiment 2

The concrete grammar of preparation acyl group sulfonamide intermediate

To (1R, 2S) 1-t-butoxycarbonyl amino-2-vinyl-cyclopropane-carboxylic acid (217mg, add in THF 1.194mmol) (5mL) solution CDI (290mg, 1.791mmol), and with reaction mixture refluxed heating 45 minutes.In another round-bottomed flask, with LiHMDS (1.0M solution, in hexane, 2.4mL, (330mg, in THF 2.388mmol) (5mL) solution, at room temperature stirred reaction mixture is 1 hour 2.4mmol) to join N-ethyl-methyl sulfonamide.Two reactant mixtures are lumped together, at room temperature stirred 2 hours.Add entry, with the quencher reaction, with EtOAc abstraction reaction solution.Separate organic facies, use MgSO ₄Dry.Evaporating solvent obtains bullion, and it with preparation HPLC purification, is obtained needed N-acyl group sulfonamide.Then N-acyl group sulfonamide is dissolved in 4N HCl De diox (2mL) solution, stirring at room 4 hours.Evaporating liquid obtains the HCl salt of hazel oil form.(112mg, 33% productive rate). ¹HNMR (400Mz, CD ₃OD) δ 1.16 (t, J=7.21Hz, 3H), 1.68 (dd, J=10.03,7.83Hz; 1H), 2.15 (m, 1H), 2.37 (m, 1H), 2.89 (s; 3H), 3.30 (m, 2H), 5.31 (d, J=10.27Hz, 1H); 5.42 (d, J=17.12Hz, 3H), 5.68 (m, 1H) ..LC-MS (retention time: 0.883min.), MS m/z 270 (M+Na ⁺).

Embodiment 3:

Raceme (1R, 2S)/(1S, 2R)-preparation (method A and method B) of 1-amino-2-vinyl cyclopropane-carboxylic acid carbethoxy hydrochloride

Utilize following method A and the racemic chemical compound of naming of each preparation among the B.

Method A

The preparation of the N-benzyl imines of glycine ethyl ester

(303.8g 2.16mole) is suspended in the t-butyl methyl ether (1.6L) with glycine ethyl ester hydrochloride.(231g, 2.16mole) (154.6g, 1.09mole), the use ice-water bath is cooled to 0 ℃ with mixture with anhydrous sodium sulfate to add benzaldehyde.With dropwise added in 30 minutes triethylamine (455mL, 3.26mole), in stirring at room mixture 48 hours.Come the quencher reaction through adding freezing water (1L) then, separate organic layer.Extract water with t-butyl methyl ether (0.5L), use saturated NaHCO ₃The organic facies that the mixture washing of aqueous solution (1L) and saline (1L) merges.Use MgSO ₄Drying solution, vacuum concentration obtains 392.4g N-benzyl imines product goldenrod oil, and it can be directly in next step use. ¹H?NMR(CDCl ₃，300MHz)δ1.32(t，J＝7.1Hz，3H)，4.24(q，J＝7.1Hz，2H)，4.41(d，J＝1.1Hz，2H)，7.39-7.47(m，3H)，7.78-7.81(m，2H)，8.31(s，?1H)。

Raceme N-Boc-(1R, 2S)/(1S, 2R)-preparation of 1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester

With 60 fens clockwise tert-butyl alcohol lithium (84.06g; 1.05mol) dry toluene (1.2L) suspension in dropwise add the N-benzyl imines (100.4g of glycine ethyl ester; 0.526mol) and anti-form-1,4-two bromo-2-butylene (107.0g, the mixture of dry toluene 0.500mol) (0.6L).After adding finished, (TBME 1L) came the dark red mixture of quencher through adding entry (1L) and t-butyl methyl ether.Water phase separated uses TBME (1L) to extract once more.Merge organic facies, add 1N HCl (1L), at room temperature stirred the mixture 2 hours.Separate organic facies, water (0.8L) extracts.Merge water then, (700g) is saturated with salt, adds TBME (1L), and mixture is cooled to 0 ℃.Alkalize to pH value 14 through dropwise adding 10N NaOH, will stirring the mixture then, separate organic layer, (2 * 500mL) extract water with TBME.With the dry (MgSO of the organic extract that merges ₄), be concentrated into the 1L volume.In this unhindered amina solution, add BOC ₂O or two dimethyl dicarbonate butyl esters (131.0g, 0.6mol), in stirring at room mixture 4 days.(50g 0.23mol) joins in the reaction, and backflow mixture 3 hours is cooled to ambient temperature overnight then with two extra dimethyl dicarbonate butyl esters.Reactant mixture is used MgSO ₄Drying, vacuum concentration obtains the 80g bullion.Through flash chromatography (2.5Kg SiO ₂, with 1% to 2%MeOH/CH ₂Cl ₂Eluting) purification residue, obtain 57g (53%) raceme N-Boc-(1R, 2S)/(1S, 2R)-1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester yellow oil, when in refrigerator, leaving standstill, it solidifies: ¹H NMR (CDCl ₃, 300MHz) δ 1.26 (t, J=7.1Hz, 3H), 1.46 (s, 9H), 1.43-1.49 (m, 1H); 1.76-1.82 (br m, 1H), 2.14 (q, J=8.6Hz, 1H), 4.18 (q, J=7.2Hz, 2H); 5.12 (dd J=10.3,1.7Hz, 1H), 5.25 (br s, 1H), 5.29 (dd, J=17.6; 1.7Hz, 1H), 5.77 (ddd, J=17.6,10.3,8.9Hz, 1H); MS m/z254.16 (M-1).

Raceme (1R, 2S)/(1S, 2R) preparation of 1-amino-2-vinyl cyclopropane-carboxylic acid carbethoxy hydrochloride

With N-Boc-(1R, 2S)/(1S, 2R)-1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester (9.39g, 36.8mmol) be dissolved in 4N HCl/ diox (90mL, 360mmol) in, stirring at room 2 hours.Concentrated reaction mixture, provide (1R, 2S)/(1S, 2R)-1-amino-2-vinyl cyclopropane-carboxylic acid carbethoxy hydrochloride, quantitative yield (7g, 100%). ¹H NMR (methanol-d ₄) δ 1.32 (t, J=7.1,3H), 1.72 (dd, J=10.2,6.6Hz, 1H), 1.81 (dd; J=8.3,6.6Hz, 1H), 2.38 (q, J=8.3Hz, 1H), 4.26-4.34 (m, 2H); 5.24 (dd, 10.3,1.3Hz, 1H) 5.40 (d, J=17.2,1H), 5.69-5.81 (m, 1H).

Method B

The preparation of raceme N-Boc-1-amino-2-vinyl cyclopropane-carboxylic acid carbethoxy hydrochloride

At-78 ℃, ((25.0g is in THF 93.53mmol) (112mL) solution for N-dibenzyl imines for 11.55g, the N of the glycine ethyl ester that adding is purchased in THF 102.9mmol) (450mL) solution to potassium tert-butoxide.Reactant mixture is heated to 0 ℃, stirred 40 minutes, be cooled to-78 ℃ then.In this solution, add anti-form-1, (20.0g's 4-two bromo-2-butylene 93.50mmol), stirred the mixture 1 hour at 0 ℃, was cooled to-78 ℃.(11.55g 102.9mmol), is heated to 0 ℃ with mixture, restir one hour, then vacuum concentration immediately to add potassium tert-butoxide.Bullion is received in Et ₂Among the O (530mL), (106mL 106mmol), stirs the biphase mixture that obtains 3.5 hours in room temperature to add 1N HCl aqueous solution.Separate each layer, use Et ₂O (2x) washs water layer, uses saturated NaHCO ₃The aqueous solution alkalization.Use Et ₂O (3x) extracts needed amine, and the organic extract that merges is used brine wash, dry (MgSO ₄), vacuum concentration obtains unhindered amina.(100mL 400mmol) handles, and concentrates with 4N HCl De dioxane solution with this material; Obtain (1R; 2S)/(1S, 2R)-1-amino-2-vinyl cyclopropane-carboxylic acid carbethoxy hydrochloride brown semi-solid (5.3g, 34% productive rate); It is identical with the material that method A obtains, and only has a small amount of unidentified fragrant impurity (8%).

Embodiment 4

N-Boc-(1R, 2S)/(1S, 2R)-fractionation of 1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester

Split A

Aqueous solution (0.1M to sodium phosphate buffer; 4.25 rise (" L "); PH value 8) (is placed in 12 liters of jacketed reactors,, stirs) the middle 511 gram Alcalase 2.4L (approximately 425mL) (Novozymes North America Inc.) that add with 300rpm 39 ℃ of maintenances.When the temperature of mixture reaches 39 ℃,, pH value is adjusted to 8.0 through adding 50%NaOH/ water.Add in then during 40 minutes raceme N-Boc-(1R, 2S)/(1S, 2R)-the 850mL DMSO solution of 1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester (85g).Then reaction temperature was kept 24.5 hours at 40 ℃, during this period, use 50%NaOH/ water, time the name a person for a particular job pH value of mixture 1.5 hours and 19.5 hours is adjusted to 8.0.24.5 after hour, measuring the mapping of ester excessive is 97.2%, with reaction cooled to room temperature (26 ℃), and stirred overnight (16 hours), then measuring the mapping of ester excessive is 100%.With 50%NaOH the pH value of reactant mixture is adjusted to 8.5 then, and (2 * 2L) extract with MTBE with the mixture that obtains.Then the MTBE extract that merges is used 5%NaHCO ₃(3 * 100mL), water (3 * 100mL) washing, vacuum evaporation, obtain enantiomer-pure N-Boc-(1R, 2S)/-1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester light yellow solid (42.55g; Purity: 97%210 nanomoles (" nM ") do not comprise acid; 100% enantiomeric excess (" ee ").

Use 50%H then ₂SO ₄The water layer that is obtained from leaching process is acidified to pH value 2, and (2 * 2L) extract with MTBE.Water (3 * 100mL) washing MTBE extracts, evaporation obtains sour light yellow solid (42.74g; Purity: 99%210nM does not comprise ester).

1R, 2S-ester 1S, 2R-acid

Split B

(the volume: the 10ml/ hole) in the hole of 24 hole plates; In 0.5mL 100 mMs (" mM ") HepsNa buffer (pH value 8.5), add 0.1mL Savinase 16.0L (protease; Be obtained from Bacillus clausii) (Novozymes North America Inc.) and raceme N-Boc-(1R; 2S)/(1S, 2R)-the 0.1mL DMSO solution of 1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester (10mg).With the plate sealing, 40 ℃, cultivation under 250rpm.After 18 hours, measuring the mapping of ester as follows excessive is 44.3%: remove the 0.1mL reactant mixture, and mix preferably with 1mL ethanol; After centrifugal, analyze 10 microlitres (" μ l ") supernatant with chirality HPLC.In remaining reactant mixture, add 0.1mL DMSO, 40 ℃, under 250rpm extra culture dish 3 days, then 4mL ethanol is joined in the hole.After centrifugal, analyze 10 μ l supernatant with chirality HPLC, the mapping of mensuration ester is excessive to be 100%.

Split C

(the volume: the 10ml/ hole) in the hole of 24 hole plates; In 0.5mL 100mM HepsNa buffer (pH value 8.5), add 0.1mL Esperase 8.0L (protease; Be obtained from Bacillushalodurans (Novozymes North America Inc.) and raceme N-Boc-(1R; 2S)/(1S, 2R)-the 0.1mL DMSO solution of 1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester (10mg).With the plate sealing, 40 ℃, cultivation under 250rpm.After 18 hours, measuring the mapping of ester as follows excessive is 39.6%: remove the 0.1mL reactant mixture, and mix preferably with 1mL ethanol; After centrifugal, analyze 10 microlitre supernatant with chirality HPLC.In remaining reactant mixture, add 0.1mL DMSO, 40 ℃, under 250rpm extra culture dish 3 days, then 4mL ethanol is joined in the hole.After centrifugal, analyze 10 μ l supernatant with chirality HPLC, the mapping of mensuration ester is excessive to be 100%.

Carry out sample analysis with following mode:

1) sample preparation: approximately the 0.5mL reactant mixture mixes with the EtOH of 10 volumes preferably.After centrifugal, 10 μ l supernatant are expelled on the HPLC post.

2) transformation assay:

Post: YMC ODS A, 4.6 * 50 millimeters (" mm "), S-5 μ m

Solvent: A, 1mM HCl/ water; B, MeCN

Gradient: 30%B, 1min; 30% to 45%B, 0.5min; 45%B, 1.5min; 45% to 30%B, 0.5min.

Flow velocity: 2ml/min

UV detects: 210nM

Retention time: acid, 1.2min; Ester, 2.8min.

3) quantitative determination is crossed in the mapping of ester:

Post: CHIRACEL OD-RH, 4.6 * 150mm, S-5 μ m

Mobile phase: MeCN/50mM HClO ₄/ water (67/33)

Flow velocity: 0.75ml/min.

UV detects: 210nM.

Retention time:

(1S, 2R)-1-amino-2-vinyl cyclopropane-carboxylic acid, 5.2min;

Raceme (1R, 2S)/(1S, 2R)-1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester, 18.5min and 20.0min;

(1R, 2S)-1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester, 18.5min.

Split D

In 20 liters of jacketed reactors, 5L 0.3M sodium phosphate buffer (pH value 8) is remained on 38 ℃, under 130rpm, stir.4 liters of Alcalase 2.4L (Novozymes North AmericaInc.) and 1 liter of DI water are joined in the reaction vessel.When the temperature of mixture during, pH value is adjusted to 7.8 with 10NNaOH near 38 ℃.During 1 hour, through add funnel with raceme N-Boc-(1R, 2S)/(1S, 2R)-5 liters of DMSO solution of 1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester (500 gram) join in the reaction vessel.Then reaction temperature is adjusted to 48 ℃.After 21 hours, the mapping of ester is excessive to reach 99.3%.Located to stop heating at 24 hours, will react and be cooled to room temperature (about 25 ℃), stirred overnight at leisure.With 10N NaOH the pH value of reactant mixture is adjusted to 8.5 then, and (2 * 4L) extract with MTBE with mixture.Use 5%NaHCO ₃(3 * 400mL) and water (the MTBE extract that 3 * 400mL) washings merge, evaporation, obtain enantiomer-pure N-Boc-(1R, 2S)/-1-amino-light yellow crystallization (259g of 2-vinyl cyclopropane carboxylic acid acetoacetic ester; Purity: 96.9%210nM does not comprise acid; 100%ee).

Split E

In 20 liters of jacketed reactors, 10L 0.1M sodium phosphate buffer (pH value 8) is remained on 40 ℃, under 360rpm, stir.1.5 liters of Alcalase 2.4L (Novozymes NorthAmerica Inc.) are joined in the reaction vessel.When the temperature of mixture during, pH value is adjusted to 8.0 with 10NNaOH near 38 ℃.During 1 hour, through add funnel with raceme N-Boc-(1R, 2S)/(1S, 2R)-2 liters of DMSO solution of 1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester (200 gram) join in the reaction vessel.Then reaction temperature is adjusted to 40 ℃.After 3 hours, pH value is adjusted to 8.0 with 10N NaOH.After 21 hours, with reaction cooled to 25 ℃.With 10N NaOH the pH value of reactant mixture is adjusted to 8.5, and (2 * 5L) extract with MTBE with mixture.Use 5%NaHCO ₃(3 * 500mL) and water (the MTBE extract that 3 * 200mL) washings merge obtains 110 gram yellow oils.With oil under vacuum condition, at room temperature place, obtain enantiomer-pure N-Boc-(1R, 2S)/-1-amino-excellent crystallization (101g of the colourless length of 2-vinyl cyclopropane carboxylic acid acetoacetic ester; Purity: 97.9%210nM does not comprise acid; 100%ee).

The N-Boc-of crystal structure enantiomer-pure (1R, 2S)/-1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester is characteristic (X ray NB#:52795-093, a numbering: 634592N1) with the monocrystalline analysis.Do not form absolute configuration, for want of known chiral centre or than heavy atom.Through the intermolecular hydrogen bonding between amide groups and the carbonylic oxygen atom, form chain structure (N...O 3.159

) along crystalline a-axle center.

N-Boc-(1R, 2S)-structure of 1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester:

Crystallization data: experimental:

Chemical formula: C13H21N1O4 Crystallization

Crystallographic system: orthorhombic crystallization source: MTBE

Space group: P2 ₁2 ₁2 ₁Crystallization explanation: colourless bar-shaped

A=5.2902 (1)

α=90 ° crystal sizes (mm): 0.12 * 0.26 * 0.30

B=13.8946 (2)

β=90 ° Data collection

C=19.9768 (3)

γ=90 ° temperature (K): 293

V＝1468.40(4)? θ _max(°)：65.2(Cu?Kα)

Z=4 d _x=1.155gcm ^-3The reflected value of being measured: 7518

The reflected value of cell parameter: the value of 6817 independent reflections: 2390 (R _Int=0.0776)

The scope of cell parameter (°): the reflected value of 2.2-65.2 observation: (I >=2:2284

Absorptance (mm ^-1): 0.700 absorbs correction (T _Min-T _Max): 0.688-1.000

Split F

In 20 liters of jacketed reactors, 5L 0.2M sodium borate buffer liquid (pH value 8) is remained on 45 ℃, under 400rpm, stir.3 liters of DI water and 4 liters of Savinase 16L (EX type) (Novozymes North America Inc.) are joined in the reaction vessel.When the temperature of mixture during, pH value is adjusted to 8.5 with 10N NaOH near 45 ℃.During 40 minutes, through add funnel with raceme N-Boc-(1R, 2S)/(1S, 2R)-2 liters of DMSO solution of 1-amino-2-vinyl cyclopropane carboxylic acid acetoacetic ester (200 gram) join in the reaction vessel.Then reaction temperature is adjusted to 48 ℃.After 2 hours, pH value is adjusted to 9.0 with 10N NaOH.At 18 hours, the mapping of ester is excessive to reach 72%, with 10N NaOH pH value is adjusted to 9.0.At 24 hours, temperature is reduced to 35 ℃.At 42 hours, temperature is risen to 48 ℃, with 10N NaOH the pH value is adjusted to 9.0.Located to stop heating at 48 hours, will react and be cooled to room temperature (about 25 ℃), stirred overnight at leisure.At 66 hours, the pH value of reactant mixture was 8.6.(2 * 4L) extract mixture with MTBE.Use 5%NaHCO ₃(3 * 300mL) and water (the MTBE extract that 3 * 300mL) washings merge, evaporation, obtain enantiomer-pure N-Boc-(1R, 2S)/-the 1-amino-light yellow crystallization of 2-vinyl cyclopropane carboxylic acid acetoacetic ester (101A g; Purity: 95.9%210nM does not comprise acid; 98.6%ee).

Embodiment 5

The preparation of step 1:1 (R)-amino-2 (S)-vinyl cyclopropane-carboxylic acid carbethoxy hydrochlorides

At N ₂In the atmosphere, in room temperature, (8.5g 33.3mmol) stirred 3 hours with 200mL 4N HCl/ diox (Aldrich) with 1 (R)-t-butoxycarbonyl amino-2 (S)-vinyl cyclopropane carboxylic acid acetoacetic ester.Removal of solvent under reduced pressure keeps temperature to be lower than 40 ℃.Obtain 6.57g (～100%) 1 (R)-hazel solid of amino-2 (S)-vinyl cyclopropane-carboxylic acid carbethoxy hydrochlorides. ¹H?NMR(300MHz，CD ₃OD)δ1.31(t，J＝7.0Hz，3H)，1.69-1.82(m，2H)，2.38(q，J＝8.8Hz，1H)，4.29(q，J＝7.0Hz，2H)，5.22(d，J＝10.3Hz，1H)，5.40(d，J＝17.2Hz，1H)，5.69-5.81(m，1H)..MS?m/z?156(M ⁺+1)。

The preparation of step 2:1 (R)-[1-tertbutyloxycarbonyl-4 (R)-hydroxyl pyrrolidine-2 (S)-carboxylic acid amides]-2 (S)-vinyl cyclopropane carboxylic acid acetoacetic esters

With Boc-L-4-hydroxyproline (N-Boc (2S; 4R)-and hydroxyproline) (10g; 43.3mmol) the 400mL dichloromethane stir serosity with N-methylmorpholine (9.3mL; 84.7mmol), HATU (19.5g, 51.3mmol) with 1 (R)-amino-2 (S)-vinyl cyclopropane-carboxylic acid carbethoxy hydrochloride (9.1g, 47.5mmol) sequential processing.Should the golden yellow homogeneous solution in room temperature, at N ₂Stirred 18 hours in the atmosphere, then vacuum concentration obtains brown oil.With it at ethyl acetate and saturated NaHCO ₃Distribute between the aqueous solution.Use the brine wash organic facies, dry (MgSO ₄), vacuum concentration obtains 15g (94%) 1 (R)-[1-tertbutyloxycarbonyl-4 (R)-hydroxyl pyrrolidine-2 (S)-carboxylic acid amides]-2 (S)-vinyl cyclopropane carboxylic acid acetoacetic ester off-white color solids: LC-MS (Xterra HPLC post: 3.0 * 50mm length.Gradient: 100% solvent orange 2 A/0% solvent B to 0% solvent orange 2 A/100% solvent B.The gradient time: 3min.The time of staying (Hold time): 1min.Flow velocity: 5mL/min.Detector wavelength: 220nM.Solvent orange 2 A: 10%MeOH/90%H ₂O/0.1%TFA.Solvent B:10%H ₂O/90%MeOH/0.1%TFA) (retention time: 2.09min), MS m/z369 (M ⁺+ 1).

The preparation of step 3:1 (R)-[4 (R)-hydroxyl pyrrolidine-2 (S)-carboxylic acid amides]-2 (S)-vinyl cyclopropane-carboxylic acid carbethoxy hydrochlorides.

(5.0g, stirring serosity 13.6mmol) was handled 3 hours with 4N HCl/ diox (20mL) with 1 (R)-[1-tertbutyloxycarbonyl-4 (R)-hydroxyl pyrrolidine-2 (S)-carboxylic acid amides]-2 (S)-vinyl cyclopropane carboxylic acid acetoacetic esters.The vacuum concentration reactant mixture obtains 4.5g (97%) 1 (R)-[4 (R)-hydroxyl pyrrolidine-2 (S)-carboxylic acid amides]-2 (S)-vinyl cyclopropane-carboxylic acid carbethoxy hydrochloride white solids: ¹HNMR (300MHz, CD ₃OD) δ 1.26 (t, J=7.14Hz, 3H), 1.46 (dd, J=9.70,5.31Hz, 1H), 1.80 (dd, J=8.23,5.31Hz; 1H), 2.00-2.15 (m, 1H), 2.18-2.30 (m, 1H), 2.45 (dd, J=13.36,7.50Hz, 1H), 3.36-3.48 (m; 1H), 4.11-4.24 (m, 2H), 4.44 (dd, J=10.25,7.68Hz, 1H), 4.58-4.65 (m, 1H); 4.84-4.94 (m, 1H), 5.17 (d, J=1.83Hz, 1H), 5.27-5.42 (m, 1H), 5.67-5.89 (m, 1H).

Embodiment 6

The preparation of cyclopropyl sulfonamide, method A and B

Method A:

, 100mL blasts gaseous ammonia in being cooled to 0 ℃ THF solution, till reaching capacity.In this solution, add the 50mL THF solution of 5g (28.45mmol) cyclopropyl sulfonyl chlorine (buying), solution is heated to ambient temperature overnight, extra stirring one day from Array Biopharma.Enriched mixture till solvent residue 1-2mL, is applied to 30g SiO ₂(with 30% to 60%EtOAc/ hexane eluting) beyond the Great Wall obtains 3.45g (100%) cyclopropyl sulfonamide white solid. ¹HNMR (methanol-d ₄) δ 0.94-1.07 (m, 4H), 2.52-2.60 (m, 1H); ¹³C NMR (methanol-d ₄) δ 5.92,33.01.

Method B:

The preparation of the step 1:N-tert-butyl group-(3-chloro) sulfonyl propyl amine

(3.0mol 315.3mL) is dissolved among the THF (2.5L) with tert-butylamine.Solution is cooled to-20 ℃.Add at leisure 3-chloropropane sulfonic acid chloride (1.5mol, 182.4mL).Reactant mixture is warming up to room temperature, stirred 24 hours.Filtering mixt, vacuum concentrated filtrate.Residue is dissolved in CH ₂Cl ₂(2.0L).The solution that obtains with 1N HCl (1.0L), water (1.0L), saline (1.0L) washing, is used Na ₂SO ₄Dry.Filter, vacuum concentration obtains faint yellow solid, and it is used the hexane crystallization, obtains product white solid (316.0g, 99%).

¹H?NMR(CDCl ₃)δ1.38(s，9H)，2.30-2.27(m，2H)，3.22(t，J＝7.35Hz，2H)，3.68(t，J＝6.2Hz，2H)，4.35(b，1H)。

Step 2: the preparation of cyclopropane sulfonic acid tert-butylamides

At-78 ℃, to the N-tert-butyl group-(3-chloro) sulfonyl propyl amine (2.14g, add in THF 10.0mmol) (100mL) solution n-BuLi (2.5M, in hexane, 8.0mL, 20.0mmol).Made reactant mixture be warmed to room temperature with 1 hour.Vacuum is removed volatile matter.(200mL distributes between 200mL) at EtOAC and water with residue.Separate organic facies, use brine wash, use Na ₂SO ₄Drying is filtered vacuum concentration.With the residue recrystallization, obtain needed product white solid (1.0g, 56%) with hexane.

¹H?NMR(CDCl ₃)δ0.98-1.00(m，2H)，1.18-1.19(m，2H)，1.39(s，9H)，2.48-2.51(m，1H)，4.19(b，1H)。

Step 3: the preparation of cyclopropyl sulfonamide

(110.0g, TFA 0.62mol) (500mL) solution at room temperature stirred 16 hours with cyclopropane sulfonic acid tert-butylamides.Vacuum is removed volatile matter.With the residue recrystallization, obtain needed product white solid (68.5g, 91%) with EtOAC/ hexane (60mL/240mL).

¹H?NMR(DMSO-d ₆)δ0.84-0.88(m，2H)，0.95-0.98(m，2H)，2.41-2.58(m，1H)，6.56(b，2H)。

Embodiment 7

The preparation of the N-tert-butyl group-(1-methyl) cyclopropyl sulfonamide.

The preparation of the step 1a:N-tert-butyl group-(3-chloro) sulfonyl propyl amine.

As stated.

The preparation of the step 1b:N-tert-butyl group-(1-methyl) cyclopropyl sulfonamide.

(4.3g 20mmol) is dissolved among the anhydrous THF (100mL), and is cooled to-78 ℃ with the N-tert-butyl group-(3-chloro) sulfonyl propyl amine.In this solution, add n-BuLi (2.5M is in hexane for 17.6mL, 44mmol) at leisure.Remove the dry ice bath, reactant mixture is warming up to room temperature in during 1.5 hours.Then mixture is cooled to-78 ℃, adds n-BuLi solution (2.5M is in hexane for 20mmol, 8mL).Reactant mixture is heated to room temperature, is cooled to-78 ℃ again in during 2 hours, add pure methyl iodide (5.68g, 40mmol) solution.Make reactant mixture be warming up to ambient temperature overnight, use saturated NH in room temperature ₄Cl (100mL) cancellation.Extract with EtOAc (100mL).With saline (100mL) washing organic facies, dry (MgSO ₄), vacuum concentration obtains yellow oil, and it is used the hexane crystallization, obtains product faint yellow solid (3.1g, 81%): ¹HNMR (CDCl ₃) δ 0.79 (m, 2H), 1.36 (s, 9H), 1.52 (m, 2H), 1.62 (s, 3H), 4.10 (bs, 1H).

The preparation of step 1c:1-methyl cyclopropyl sulfonamide

(1.91g, 10mmol) solution was dissolved among the TFA (30mL), stirring at room reactant mixture 16 hours with the N-tert-butyl group-(1-methyl) cyclopropyl sulfonamide.Solvent removed in vacuo obtains yellow oil, with its with the EtOAc/ hexane (1: 4,40mL) crystallization obtains embodiment 3,1-methyl cyclopropyl sulfonamide white solid (1.25g, 96%): ¹H NMR (CDCl ₃) δ 0.84 (m, 2H), 1.41 (m, 2H), 1.58 (s, 3H), 4.65 (bs, 2H) .C ₄H ₉NO ₂The analytical calculation value of S: C, 35.54; H, 6.71; N, 10.36.Measured value: C, 35.67; H, 6.80; N, 10.40.

Embodiment 9

The preparation of 1-propyl group cyclopropyl sulfonamide

The preparation of the step 1b:N-tert-butyl group-(1-benzyl) cyclopropyl-sulfonamide.

Use the described method of preparation 1-methyl cyclopropyl sulfonamide, prepare this chemical compound, just in second step of this method, use propyl halide to replace methyl iodide.

Embodiment 10

The preparation of the N-tert-butyl group-(1-pi-allyl) cyclopropyl sulfonamide.

According to the method described in the synthetic N-tert-butyl group-(1-methyl) cyclopropyl sulfonamide, just make electrophilic reagent with 1.25 normal allyl bromide, bromoallylenes, obtain this compound N-tert-butyl group-(1-pi-allyl) cyclopropyl sulfonamide with 97% productive rate.This chemical compound just can directly adopt in next one reaction without purification: ¹H NMR (CDCl ₃) δ 0.83 (m, 2H), 1.34 (s, 9H), 1.37 (m, 2H), 2.64 (d, J=7.3Hz, 2H), 4.25 (bs, 1H), 5.07-5.10 (m, 2H), 6.70-6.85 (m, 1H).

The preparation of 1-pi-allyl cyclopropyl sulfonamide.

Method according to described in 1-methyl cyclopropyl sulfonamide synthetic obtains this chemical compound 1-pi-allyl cyclopropyl sulfonamide, productive rate 40% by the N-tert-butyl group-(1-pi-allyl) cyclopropyl sulfonamide.Use SiO ₂The purified chemical compound uses 2%MeOH/CH ₂Cl ₂As eluent: ¹HNMR (CDCl ₃) δ 0.88 (m, 2H), 1.37 (m, 2H), 2.66 (d, J=7.0Hz, 2H), 4.80 (s, 2H), 5.16 (m, 2H), 5.82 (m, 1H); ¹³C NMR (CDCl ₃) δ 11.2,35.6,40.7,119.0,133.6.

Embodiment 15

Cyclopropyl sulfonyl amine tert-butyl group carbamate (preparation C ₁Key intermediate in the-substituted cyclopropyl sulfonamide) preparation

The preparation of step 1:3-chloropropyl sulfonamide

(55g, 310.7mmol) solution is dissolved among the THF (200mL), and with dropwise joining the NH that is cooled to 0 ℃ in 30 minutes with 3-chloropropane sulfonic acid chloride ₄In the OH solution (200mL).Reactant mixture is heated to room temperature, stirred 1 hour, with dichloromethane (4 * 500mL) repeated dispensing water layers.With the dichloromethane layer that 1N HCl (150mL), water (150mL) washing merge, use MgSO ₄Drying is filtered vacuum concentration.Dichloromethane/hexane recrystallization bullion solid with minimum flow obtains 3-chloropropyl sulfonamide white solid (45.3g, 93%). ¹H?NMR(CDCl ₃)δ2.34(m，2H)，3.32(t，J＝7.3Hz，2H)，3.70(t，J＝6.2Hz，2H)，4.83(s，2H)； ¹³CNMR(CDCl ₃)δ27.10，42.63，52.57。

The preparation of step 2:3-chloropropyl sulfonylcarbamic acid tertiary butyl ester

Be cooled to 0 ℃ 3-chloropropyl sulfonamide (30.2g with 30 fens clockwise; 191.5mmol), triethylamine (30.2mL; 217.0mmol) and 4-DMAP (2.40g; 19.6mmol) dichloromethane (350mL) solution in dropwise add two dimethyl dicarbonate butyl esters (47.2g, dichloromethane 216.9mmol) (250mL) solution at leisure.Make reactant mixture be warming up to room temperature, extra stirring 3 hours distributes with 1N HCl (300mL), water (300mL), saline (300mL), uses MgSO ₄Drying is filtered, and vacuum concentration obtains bullion.Grind this material with 70mL 5% dichloromethane/hexane, obtain 3-chloropropyl sulfonylcarbamic acid tertiary butyl ester pale solid (47.2g, 96%): ¹H NMR (CDCl ₃) δ 1.51 (s, 9H), 2.33 (m, 2H), 3.60 (t, J=7.3Hz, 2H), 3.68 (t, J=6.21Hz, 2H); ¹³C NMR (CDCl ₃) δ 26.50,27.95,42.37,50.40,84.76,149.53.

Step 3: the preparation of cyclopropyl sulfonylcarbamic acid tertiary butyl ester

In argon atmospher, n-BuLi (1.6M is in hexane for 74.7mL, 119.5mmol) solution is dissolved among the anhydrous THF (105mL), and is cooled to-78 ℃.With 20-30 minute, in this solution, dropwise add 3-chloropropyl sulfonylcarbamic acid tertiary butyl ester (14g, anhydrous THF solution 54.3mmol) (105mL).Remove the dry ice bath, reactant mixture is warming up to room temperature in during 2 hours.With glacial acetic acid (3.4mL) cancellation reactant mixture, vacuum concentration distributes between dichloromethane (100mL) and water (100mL).With saline (100mL) washing organic facies, dry (MgSO ₄), filtering, vacuum concentration obtains the waxy off-white color solid of cyclopropyl sulfonylcarbamic acid tertiary butyl ester (12.08g, 100%): ¹H NMR (CDCl ₃) δ 1.10 (m, 2H), 1.34 (m, 2H), 1.50 (s, 9H), 2.88 (m, 1H), 7.43 (s, 1H) .. ¹³C NMR (CDCl ₃) δ 6.21,28.00,31.13,84.07,149.82.

Embodiment 16

The preparation of 1-methoxyl group-methyl cyclopropyl sulfonamide

The preparation of step 1:1-methoxy cyclopropyl sulfonylcarbamic acid tertiary butyl ester

To be cooled to-78 ℃, (1.0g 4.5mmol) adds n-BuLi (1.6M is in hexane for 6.4mL, 10.2mmol) in the solution, and stirred reaction mixture 1 hour to be dissolved in the cyclopropyl sulfonylcarbamic acid tertiary butyl ester of THF (30mL).(0.40mL 5.24mmol), makes mixture be warming up to ambient temperature overnight to the pure solution of adding chloromethyl methyl ether at leisure in this solution.Use 1N HCl aqueous solution that the solution pH value is adjusted to 3, use ethyl acetate (4 * 50mL share) to extract then.With the dry (MgSO of the extract that merges ₄), filter, concentrate, obtain the waxy solid of 1-methoxy cyclopropyl sulfonylcarbamic acid tertiary butyl ester (1.20g, 100%), it just need not be further purified and can in next one reaction, directly adopt: ¹H NMR (CDCl ₃) δ 1.03 (m, 2H), 1.52 (s, 9H), 1.66 (m, 2H), 3.38 (s, 3H), 3.68 (s, 2H), 7.54 (s, 1H); ¹³CNMR (CDCl ₃) δ 11.37,28.29,40.38,58.94,73.43,83.61,149.57.

The preparation of step 2:1-methoxy cyclopropyl sulfonamide

(1.14g, 4.30mmol) solution is dissolved in 50%TFA/ dichloromethane (30mL) solution, at room temperature stirs 16 hours with 1-methoxy cyclopropyl sulfonylcarbamic acid tertiary butyl ester.Solvent removed in vacuo is used 80g SiO with residue ₂Chromatographic isolation (with 0% to 60% ethyl acetate/hexane eluting) obtains 1-methoxy cyclopropyl sulfonamide white solid (0.55g, 77%, two step): ¹H NMR (CDCl ₃) δ 0.95 (m, 2H), 1.44 (m, 2H), 3.36 (s, 3H), 3.65 (s, 2H), 4.85 (s, 2H); ¹³C NMR (CDCl ₃) δ 11.17,40.87,59.23,74.80; LRMS m/z183 (M ⁺+ NH ₄).

Embodiment 17

The preparation of 1-cyclopropyl methyl cyclopropyl sulfonamide

The preparation of step 1:1-cyclopropyl methyl cyclopropyl sulfonylcarbamic acid tertiary butyl ester

According to the method described in the synthetic 1-methoxy cyclopropyl sulfonylcarbamic acid tertiary butyl ester, just make electrophilic reagent with 1.10 normal cyclopropyl methyl bromides, obtain 1-cyclopropyl methyl cyclopropyl sulfonylcarbamic acid tertiary butyl ester with 92% productive rate.This chemical compound just can directly adopt in next one reaction without purification: ¹H NMR (CDCl ₃) δ 0.10 (m, 2H), 0.51 (m, 2H), 0.67 (m, 1H), 1.10 (m, 2H), 1.49 (s, 9H), 1.62 (m, 2H), 1.87 (d, J=7.0Hz, 2H).

The preparation of step 2:1-cyclopropyl methyl cyclopropyl sulfonamide

Method according to described in 1-methoxy cyclopropyl sulfonamide synthetic obtains this chemical compound, productive rate 65% by 1-cyclopropyl methyl cyclopropyl sulfonylcarbamic acid tertiary butyl ester.Use SiO ₂The purified chemical compound, use 0% to 60% ethyl acetate/hexane as eluent: ¹HNMR (CDCl ₃) δ 0.15 (m, 2H), 0.51 (m, 2H), 1.01 (m, 2H), 1.34 (m, 3H), 1.86 (d, J=7.0Hz, 2H), 4.83 (s, 2H); ¹³C NMR (CDCl ₃) δ 4.65,7.74,11.26,35.62,41.21; LRMS m/z 193 (M ⁺+ NH ₄).

Embodiment 19

The preparation of 1-(3,5-dimethyl isoxazole-4-yl) carbamoyl cyclopropane sulfonamide

The preparation of step 1:1-(3,5-dimethyl isoxazole-4-yl) carbamoyl cyclopropane sulfonylcarbamic acid tertiary butyl ester

According to the method described in the synthetic 1-methoxy cyclopropyl sulfonylcarbamic acid tertiary butyl ester, only to use 1.20 normally 3,5-dimethyl isoxazole-4-isocyanates is made electrophilic reagent, obtains the bullion chemical compound with 100% productive rate.This chemical compound just can directly adopt in next one reaction without purification.

The preparation of step 2:1-(3,5-dimethyl isoxazole-4-yl) carbamoyl cyclopropane sulfonamide

By 1.62g (4.52mmol) 1-(3; 5-dimethyl isoxazole-4-yl) carbamoyl cyclopropane sulfonamide tert-butyl group carbamate; Use the 4N HCl/ diox of 30mL (120mmol), stirred overnight concentrates; With the chromatographic separation of Biotage 40M post (with 0% to 5% ethanol/methylene eluting), (580mg) obtains this chemical compound with 50% productive rate: ¹H NMR (methanol-d ₄) δ 1.57 (m, 2H), 1.61 (m 2H), 2.15 (s, 3H), 2.30 (s, 3H), 4.84 (s, 3H); ¹³C NMR (methanol-d ₄) δ 9.65,10.94,15.01,46.11,114.82,159.45,165.55,168.15; LRMS m/z260 (M ⁺+ H).

Embodiment 20

Prepare the cyclobutyl sulfonamide by the cyclobutyl bromine

To the 30mL anhydrous diethyl ether (Et that is cooled to-78 ℃ 5.0g (37.0mmol) cyclobutyl bromine ₂O) add the pentane solution of 44mL (74.8mmol) 1.7M tert-butyl lithium in the solution, solution was heated to-35 ℃ at leisure with 1.5 hours.This mixture is joined through conduit in the 100mL hexane solution (being cooled to-40 ℃) of the new distillatory sulfonic acid chloride of 5.0g (37.0mmol) at leisure, be heated to 0 ℃ with 1 hour, carefully vacuum concentration.This mixture is dissolved in Et again ₂Among the O, with some freezing water washings once, dry (MgSO ₄), concentrate carefully.This mixture is dissolved among the 20mL THF again, dropwise joins the saturated NH of 500mL ₃Among/the THF, stirred overnight.Vacuum concentrated mixture obtains the bullion yellow solid, with the CH of minimum flow ₂Cl ₂/ hexane (contain 1-2 and drip MeOH) recrystallization obtains 1.90g (38%) cyclobutyl sulfonamide white solid. ¹HNMR (CDCl ₃) δ 1.95-2.06 (m, 2H), 2.30-2.54 (m, 4H), 3.86 (p, J=8Hz, 1H), 4.75 (brs, 2H); ¹³C NMR (CDCl ₃) δ 16.43,23.93,56.29.HRMSm/z (M-H) ^-C ₄H ₈NSO ₂Value of calculation: 134.0276, measured value 134.0282.

Embodiment 21

The preparation of cyclopenta sulfonamide

The ethereal solution of 18.5mL (37.0mmol) 2M cyclopenta-magnesium chloride is dropwise joined in the 100mL hexane solution (being cooled to-78 ℃) of the new distillatory sulfonic acid chloride of 3.0mL (37.0mmol) (being obtained from Aldrich).With 1 hour with mixture heated to 0 ℃, vacuum concentration carefully then.This mixture is dissolved in Et again ₂Among the O (200mL), with some freezing water (200mL) washings once, dry (MgSO ₄), concentrate carefully.This mixture is dissolved among the 35mL THF again, dropwise joins the saturated NH of 500mL ₃Among/the THF, stirred overnight.Vacuum concentrated mixture obtains the bullion yellow solid, through 50g filtered through silica gel residue, uses the 70%EtOAc-hexane as eluent, then concentrated solution.CH with minimum flow ₂Cl ₂/ hexane (contain 1-2 and drip MeOH) obtains 2.49g (41%) cyclopenta sulfonamide white solid with the residue recrystallization. ¹H?NMR(CDCl ₃)δ1.58-1.72(m，2H)，1.74-1.88(m，2H)，1.94-2.14(m，4H)，3.48-3.59(m，1H)，4.80(bs，2H)； ¹³C?NMR(CDCl ₃)δ25.90，28.33，63.54；MS?m/e148(M-H) ^-。

Embodiment 22

The preparation of cyclohexyl sulfonamide

The ethereal solution of 18.5mL (37.0mmol) 2M cyclohexyl chlorination magnesium (TCI Americas) is dropwise joined in the 100mL hexane solution (being cooled to-78 ℃) of the new distillatory sulfonic acid chloride of 3.0mL (37.0mmol).With 1 hour with mixture heated to 0 ℃, vacuum concentration carefully then.This mixture is dissolved in Et again ₂Among the O (200mL), with some freezing water (200mL) washings once, dry (MgSO ₄), concentrate carefully.This mixture is dissolved among the 35mL THF again, dropwise joins the saturated NH of 500mL ₃Among/the THF, stirred overnight.Vacuum concentrated mixture obtains the bullion yellow solid, through 50g filtered through silica gel residue, uses the 70%EtOAc-hexane as eluent, concentrates then.CH with minimum flow ₂Cl ₂/ hexane (contain 1-2 and drip MeOH) obtains 1.66g (30%) cyclohexyl sulfonamide white solid with the residue recrystallization: ¹H NMR (CDCl ₃) δ 1.11-1.37 (m, 3H), 1.43-1.56 (m, 2H), 1.67-1.76 (m, 1H), 1.86-1.96 (m, 2H), 2.18-2.28 (m, 2H), 2.91 (tt, J=12,3.5Hz, 1H), 4.70 (bs, 2H); ¹³CNMR (CDCl ₃) δ 25.04,25.04,26.56,62.74; MS m/e 162 (M-1) ^-

Embodiment 23

The preparation of neopentyl sulfonamide

According to the method for preparing cyclohexyl sulfonamide, 49mL (37mmol) 0.75M neopentyl magnesium chloride (Alfa) (in diethyl ether) is changed into the neopentyl sulfonamide white solid of 1.52g (27%). ¹H NMR(CDCl ₃)δ1.17(s，9H)，3.12(s，2H)，4.74(brs，2H)； ¹³CNMR(CDCl ₃)δ29.46，31.51，67.38；MS?m/e?150(M-1) ^-。

Embodiment 24

The preparation of cyclobutylmethyl sulfonamide

The 150mL acetone soln backflow of 12.3g (83mmol) cyclobutylmethyl bromine (Aldrich) and 13.7g (91mmol) sodium iodide is spent the night, then be cooled to room temperature.Leach inoganic solids, respectively at ambient temperature and 150torr, distill out acetone and cyclopropyl methyl iodide (8.41g, 46%) at 80 ℃.

30mL anhydrous diethyl ether (diethyl ether) solution that is cooled to-78 ℃ 4.0g (21.98mmol) cyclobutylmethyl iodine is joined in the cyclohexane solution of 17mL (21.98mmol) 1.3M s-butyl lithium through conduit, stirred this solution 5 minutes.The 110mL hexane solution that in this mixture, adds the new distillatory sulfonic acid chloride of 3.0g (21.98mmol) be cooled to-78 ℃ through conduit, with 1 hour with this mixture heated to room temperature, vacuum concentration carefully then.This mixture is dissolved in the diethyl ether again, with some freezing water washings once, dry (MgSO ₄), filter, concentrate carefully.This mixture is dissolved among the 30mL THF again, dropwise joins the saturated NH of 500mL ₃Among/the THF, stirred overnight.Vacuum concentrated mixture obtains the bullion yellow solid, with dichloromethane/hexane (contain 1-2 and the drip methanol) recrystallization of minimum flow, obtains 1.39g (42%) cyclobutylmethyl sulfonamide white solid. ¹H?NMR(CDCl ₃)δ1.81-2.03(m，4H)，2.14-2.28(m，2H)，2.81-2.92(m，1H)，3.22(d，J＝7Hz，2H)，4.74(brs，2H)； ¹³C?NMR(CDCl ₃)δ19.10，28.21，30.64，60.93；MS?m/e?148(M-1) ^-。

Embodiment 25

The preparation of cyclopropyl carbinyl sulfonamide

Use the method for preparing the cyclobutylmethyl sulfonamide, prepare the cyclopropyl carbinyl sulfonamide (referring to JACS 1981, p.442-445) by cyclopropyl carbinyl bromine (Aldrich). ¹H?NMR(CDCl ₃)δ0.39-0.44(m，2H)，0.67-0.76(m，2H)，1.13-1.27(m，1H)，3.03(d，J＝7.3Hz，2H)，4.74(brs，2H)； ¹³C?NMR(CDCl ₃)δ4.33，5.61，59.93；MS?m/e134(M-1)。

Embodiment 26

The preparation of 2-thienyl sulphonyl amine

Use Justus Liebigs Ann.Chem., 501,1933, method p.174-182 is by 2-thienyl sulphonyl chlorine (buying from Aldrich) preparation.

Embodiment 27

The preparation of 4-bromobenzene sulfonamide

Handle the 4-bromine sulfonic acid chloride that is purchased with saturated ammonia/THF, preparation 4-bromophenyl sulfonamide.

Embodiment 28

The preparation of cyclopropane sulfonic acid (1-(R)-amino-2-(S)-vinyl-cyclopropane carbonyl) amide HCl salt

The preparation of step 1:1 (R)-t-butoxycarbonyl amino-2 (S)-vinyl-cyclopropane-carboxylic acid

(3.28g adds LiOH (1.27g, water 53.0mmol) (14mL) suspension in THF 13.2mmol) (7mL) and methanol (7mL) solution to 1 (R)-t-butoxycarbonyl amino-2 (S)-vinyl-cyclopropane carboxylic acid acetoacetic ester.At room temperature stir the mixture and spend the night, with 1N NaOH (15mL) and water (20mL) cancellation.Mixture with ethyl acetate (20mL) washing obtains extracts organic facies with 20mL 0.5NNaOH.With the water-soluble liquid phase that 1N HCl acidify merges, till pH value 4, (3 * 40mL) extract with ethyl acetate.With the organic extract that brine wash merges, dry (MgSO ₄), filter, concentrate, obtain title compound white solid (2.62g, 87%). ¹HNMR: (DMSO-d ₆) δ 1.22-1.26 (m, 1H), 1.37 (s, 9H), 1.50-1.52 (m, 1H), 2.05 (q, J=9Hz; 1H), 5.04 (d, J=10Hz, 1H), 5.22 (d, J=17Hz, 1H); 5.64-5.71 (m, 1H), 7.18,7.53 (s, NH (rotamer), 12.4 (br s, 1H)); MS m/z228 (M ⁺+ H).

Step 2: the preparation of cyclopropane sulfonic acid (1-(R)-t-butoxycarbonyl amino-2-(S)-vinyl cyclopropane carbonyl)-amide

In nitrogen atmosphere, with step 1 product (2.62g, 11.5mmol) and CDI (2.43g, THF 15.0mmol) (40mL) solution reflux 50 minutes.This solution is cooled to room temperature, and (1.82g is in THF 15.0mmol) (10mL) solution to transfer to the cyclopropyl sulfonamide through sleeve pipe.(2.40mL 16.1mmol), continues to stir 20 hours in the solution that obtains, to add DBU.With 1N HCl cancellation mixture, to pH value 1, vacuum concentration THF.(2 * 50mL) extract suspension, dry (Na with ethyl acetate ₂SO ₄) organic extract that merges, filter, concentrate.Through carrying out purification, obtain title compound (2.4g) white solid with hexane-ethyl acetate (1: 1) recrystallize.Through Biotage 40S column purification mother solution (9% acetone/dichloromethane eluting), obtain second batch of title compound (1.1g).Merge two batch of materials (total recovery 92%). ¹H NMR (DMSO-d ₆) δ 0.96-1.10 (m, 4H), 1.22 (dd, J=5.5,9.5Hz, 1H), 1.39 (s, 9H), 1.70 (t; J=5.5Hz, 1H), 2.19-2.24 (m, 1H), 2.90 (m, 1H), 5.08 (d, J=10Hz, 1H); 5.23 (d, J=17Hz, 1H), 5.45 (m, 1H), 6.85,7.22 (s, NH (rotamer); MS m/z 331 (M ⁺+ H).

Step 3: the preparation of cyclopropane sulfonic acid (1-(R)-amino-2-(S)-vinyl-cyclopropane carbonyl) amide HCl salt

(3.5g, dichloromethane 10.6mmol) (35mL) and TFA (32mL) solution at room temperature stirred 1.5 hours with step 2 product.Vacuum is removed volatile matter, residue is suspended in the 1N HCl/ diethyl ether (20mL) vacuum concentration.This method repeats once.Grind the mixture that obtains with pentane, filter, obtain the hygroscopic off-white color solid of title compound (2.60g, 92%). ¹HNMR：(DMSO-d ₆)δ1.01-1.15(m，4H)，1.69-1.73(m，1H)，1.99-2.02(m，?1H)，2.38(q，J＝9Hz，1H)，2.92-2.97(m，1H)，5.20(d，J＝11Hz，1H)，5.33(d，J＝17Hz，1H)，5.52-5.59(m，1H)，9.17(br?s，3H)；MS?m/z?231(M ⁺+H)。

Embodiment 29

(1S, 4R, 6S, 14S, 18R)-and 7-cis-14-t-butoxycarbonyl amino-18-hydroxyl-2,15-dioxo-3,16-diaza tricyclic [14.3.0.0 ^4,6] preparation of 19-7-alkene-4-carboxylic acid, embodiment 29

The preparation of step 1:1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (S)-carboxylic acid methyl ester

With 2 (S)-t-butoxycarbonyl amino-8-nonenoic acid (buying) (3.5g from RSP Amino Acids; 12.9mmol) 200mL DCM solution with 4 (R)-hydroxyl pyrrolidines-2 (S)-carboxylic acid methyl ester hydrochloride (2.15g; 11.8mmol), N-methylmorpholine (4.25mL; 38.6mmol) and HATU (5.37g, 14.1mmol) sequential processing.In room temperature, at N ₂Stirred reaction mixture is 3 days in the atmosphere, then vacuum concentration.Residue is distributed between the buffer (biphthalate) of ethyl acetate and pH value 4.Use saturated NaHCO ₃The solution washing organic facies, dry (MgSO ₄), vacuum concentration obtains bullion.Carry out flash chromatography (50% ethyl acetate/hexane to 100% ethyl acetate), obtain 1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (the S)-carboxylic acid methyl ester water white oils of 4.7g (～100%): ¹H NMR (500MHz, CD ₃OD) δ 1.33-1.50 (m, 8H), 1.46 (s, 9H), 1.57 (m, 1H), 1.72 (m, 1H) 2.08 (m; 2H), 2.28 (m, 1H), 3.72 (s, 3H) 3.75-3.87 (m, 2H); 4.36 (m, 1H), 4.51 (bs, 1H), 4.57 (t, J=8.2Hz, 1H); 4.95 (d, J=10.4Hz, 1H), 5.01 (m, 1H), 5.83 (m, 1H); MS m/z399 (M ⁺+ 1).

Step 2:1-{ [1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (S) carbonyls]-(1R)-amino } preparation of-2 (S)-vinyl-cyclopropane carboxylic acid acetoacetic ester

(4.7g 11.8mmol) is dissolved in THF (80mL), methanol (20mL) and the water (40mL) with 1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (S)-carboxylic acid methyl ester.Adding powder Lithium hydrate (5.6g, 233mmol).In room temperature, at N ₂Stirred pale yellow syrup 16 hours in the atmosphere, then vacuum concentration.Residue is distributed between ether and water.Remove the ether phase, handle water with 1N HCl, till pH value 4.Extract this acid solution with EtOAc (3x).With the dry (MgSO of the EtOAc extract that merges ₄), vacuum concentration obtains 4.36g (96%) 1-(2 (S)-t-butoxycarbonyl amino-8-nonene acyl group)-4 (R)-hydroxyl-pyrrolidine-2 (S)-carboxylic acid white solids.Then this acid is dissolved among the DMF of 150mL, add (1R, 2S)-1-amino-2-vinyl cyclopropane-carboxylic acid carbethoxy hydrochloride (2.61g, 13.6mmol), N-methylmorpholine (2.5mL, 22.6mmol) and HATU (5.2g, 13.7mmol).In room temperature, at N ₂Stirred reaction mixture is 16 hours in the atmosphere, then vacuum concentration.Residue is distributed between the buffer (biphthalate) of ethyl acetate and pH value 4.Use saturated NaHCO ₃The solution washing organic facies, dry (MgSO ₄), vacuum concentration obtains bullion.Carry out flash chromatography (60%-80% ethyl acetate/hexane), obtain 6.0g (98%) 1-{ [1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (S) carbonyls]-(1R)-amino-2 (S)-vinyl-cyclopropane carboxylic acid acetoacetic ester white solid: ¹HNMR (500MHz, CD ₃OD) δ 1.25 (t, J=7.2Hz, 3H), 1.33-1.80 (m, 10H), 1.46 (s, 9H), 2.09 (m, 3H), 2.25 (m; 2H), 3.76 (m, 2H), 4.14 (m, 2H), 4.27 (dd, J=8.5,5.2Hz, 1H), 4.50 (m; 2H), 4.94 (d, J=10.1Hz, 1H), 5.01 (dd, J=17.1,1.8Hz, 1H), 5.11 (dd, J=10.4; 1.8Hz, 1H), 5.30 (d, J=15.6Hz, 1H), 5.80 (m, 2H), 8.57 (s, 1H); MS m/z 522 (M ⁺+ 1).

Step 3: (1S, 4R, 6S, 14S, 18R)-and 7-cis-14-t-butoxycarbonyl amino-18-hydroxyl-2,15-dioxo-3,16-diaza tricyclic [14.3.0.0 ^4,6] preparation of 19-7-alkene-4-carboxylic acid, ethyl ester

With 1-{ [1-(2 (S)-tertbutyloxycarbonyl-amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (S) carbonyls]-(1R)-amino } (800mg, 2L dichloromethane solution 1.53mmol) is used N to-2 (S)-vinyl cyclopropane-carboxylic acid, ethyl esters ₂Purged 0.5 hour.(Strem) (64mg 0.075mmol), uses N to [benzal]-ruthenous chloride (IV) to add tricyclohexyl phosphine [1, two (2,4,6-trimethyl-phenyl)-4 of 3-, 5-glyoxalidine-2-subunit] then ₂Purged mixture again 10 minutes.Should reflux 2 hours by greenish orange color homogeneous solution, obtain dark orange solution.Reactant mixture is cooled to room temperature, and vacuum concentration obtains orange oil.Carry out flash chromatography (ethyl acetate), obtain 460mg (61%) (1S, 4R, 6S, 14S, 18R)-7-cis-14-t-butoxycarbonyl amino-18-hydroxyl-2,15-dioxo-3,16-diaza tricyclic [14.3.0.0 ^4,6]-19-7-alkene 4-carboxylic acid, ethyl ester gray solid. ¹HNMR(500MHz，CDCl ₃)δ1.19(t，J＝7.2Hz，3H)，1.42(s，9H)，1.22-1.8(m，8H)，1.87(m，2H)，2.03-2.22(m，4H)，2.63(m，1H)，3.65(m，1H)，4.09(m，3H)，4.45(m，1H)，4.56(s，1H)，4.82(m，1H)，5.23(m，1H)，5.51(s，1H)，7.16(s，1H)；MS?m/z?494(M ⁺+1)。

Step 4: (1S, 4R, 6S, 14S, 18R)-and 7-cis-14-t-butoxycarbonyl amino-18-hydroxyl-2,15-dioxo-3,16-diaza tricyclic [14.3.0.0 ^4,6]-19-7-alkene-4-carboxylic acid

To (1S, 4R, 6S, 14S, 18R)-and 7-cis-14-t-butoxycarbonyl amino-18-hydroxyl-2,15-dioxo-3,16-diaza tricyclic [14.3.0.0 ^4,6]-19-7-alkene-4-carboxylic acid, ethyl ester (493mg, adding powder Lithium hydrate in THF 1.0mmol) (4mL), methanol (1mL) and water (2mL) solution (480mg, 20mmol), and in room temperature, at N ₂Stirred this pale yellow syrup 16 hours in the atmosphere.Vacuum concentrated mixture then, and residue distributed between ether and water.Remove the ether phase, handle water with 1N HCl, till pH value 4.Extract this acid solution three times with EtOAc.With the dry (MgSO of the EtOAc extract that merges ₄), vacuum concentration obtains 460mg (98%) embodiment 18, (1S, 4R, 6S, 14S, 18R)-and 7-cis-14-t-butoxycarbonyl amino-18-hydroxyl-2,15-dioxo-3,16-diaza tricyclic [14.3.0.0 ^4,6]-19-7-alkene-4-carboxylic acid gray solid. ¹HNMR(500MHz，CD ₃OD)δppm?1.26(t，J＝7.2Hz，3H)，1.35-1.52(m，15H)，1.57-1.68(m，3H)，1.79(m，1H)，2.04(m，1H)，2.16-2.41(m，3H)，3.80(dd，J＝10.7，4.3Hz，1H)，3.88(m，1H)，4.38(dd，J＝8.9，3.1Hz，1H)，4.55(m，2H)，5.39(t，J＝9.8Hz，1H)，5.58(m，1H)；MS?m/z?466(M ⁺+1)。

Embodiment 30

(4-cyclopropane sulfonyl amino carbonyl-18-hydroxyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl)-preparation of carbamic acid tertiary butyl ester

Step 1:1-{ [1-(2-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4-(tert-butyl group-dimethyl-silanyloxy base)-pyrrolidine-2-carbonyl]-amino }-preparation of 2-vinyl cyclopropane carboxylic acid acetoacetic ester

To 1-{ [1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (S) carbonyls]-(1R)-amino }-2 (S)-vinyl-cyclopropane carboxylic acid acetoacetic ester (1.5g; 2.87mmol) the 10mLDMF mixture in add imidazoles (0.25g; 3.67mmol) and the tert-butyl group-dimetylsilyl chlorine (516mg, 3.44mmol).With mixture stirring at room two days.The vacuum concentration reactant mixture is dissolved in residue in the ethyl acetate then.With this solution of water washing, use dried over mgso, vacuum concentration obtains the bullion solid.With purification by flash chromatography (with 20% ethyl acetate/hexane eluting), obtain 1.43g (78%) 1-{ [1-(2-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4-(tert-butyl group-dimethyl-silanyloxy base)-pyrrolidine-2-carbonyl]-amino }-2-vinyl cyclopropane carboxylic acid acetoacetic ester white solid.

¹H?NMR(300MHz，CD ₃OD)δ0.10(s，6H)，0.89(s，9H)，1.22(m，3H)，1.31-1.48(m，16H)，1.50-1.75(m，3H)，2.06(m，3H)，2.11-2.33(m，2H)，3.70(m，2H)，4.03-4.19(m，2H)，4.21(m，1H)，4.45(t，J＝7.87Hz，1H)，4.59(m，1H)，4.91(d，J＝9.15Hz，1H)，4.98(d，J＝17.20Hz，1H)，5.08(dd，J＝10.25，1.83Hz，1H)，5.27(dd，J＝17.38，1.65Hz，1H)，5.65-5.87(m，2H)；MS?m/z?636(M ⁺+1)。

Step 2:14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] preparation of 19-7-alkene-4-carboxylic acid, ethyl ester

To 1-{ [1-(2-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4-(tert-butyl group-dimethyl-silanyloxy base)-pyrrolidine-2-carbonyl]-amino }-2-vinyl-cyclopropane carboxylic acid acetoacetic ester (1.63g; 2.56mmol) the 640mL dichloromethane solution in add 215mg (0.26mmol) tricyclohexyl phosphine [1; 3-two (2; 4,6-three [benzal] ruthenous chloride (IV).With mixture reflux 15 minutes.The vacuum concentration residue is then used purification by flash chromatography, with 30% ethyl acetate/hexane eluting.For sample is further decoloured, once more with the bullion chromatographic isolation,, obtain 1.5g (96%) 14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 with 50% ether/hexane eluting ^4,6] 19-7-alkene-4-carboxylic acid, ethyl ester white solid. ¹HNMR(500MHz，CD ₃Cl)δ0.06(s，3H)，0.07(s，3H)，0.86(s，9H)，1.18-1.24(m，6H)，1.34-1.64(m，14H)，1.86-1.96(m，3H)，2.02-2.09(m，1H)，2.11-2.17(m，1H)，2.19-2.28(m，1H)，2.57-2.63(m，1H)，3.50-3.54(m，1H)，3.71(dd，J＝10.22，6.26Hz，1H)，4.06-4.17(m，2H)，4.52-4.58(m，2H)，4.75(d，J＝8.55Hz，1H)，5.21(t，J＝9.92Hz，1H)，5.35(d，J＝7.63Hz，1H)，5.45-5.50(m，1H)，6.94(s，1H)；MS?m/z?608(M ⁺+1)。

Step 3:14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3, the preparation of 16-diaza-three ring [14.3.0.04,6] 19-7-alkene-4-carboxylic acid

To 14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-4-carboxylic acid, ethyl ester (1.5g, and adding powder lithium hydroxide monohydrate in the solution of THF 2.47mmol) (4mL), methanol (1mL) and water (2mL) mixed solvent system (1.0g, 50mmol).In room temperature, at N ₂Stirred pale yellow syrup 4 hours in the atmosphere.Vacuum concentrated mixture then, and residue distributed between ether and water.Remove the ether phase, handle water with 1N HCl, till pH value 4.Extract this acid solution (three times) with EtOAc.With the dry (MgSO of the EtOAc extract that merges ₄), vacuum concentration obtains 1.2g (84%) 14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-4-carboxylic acids white solid. ¹H?NMR(300?MHz，CD ₃OD)0.12(s，6H)，0.89(s，9H)，1.23-1.64(m，17H)，1.70-1.87(m，1H)，1.90-2.49(m，6H)，3.70-3.80(m，1H)，3.83-3.90(m，1H)，4.28-4.36(m，1H)，4.47-4.55(m，1H)，4.65(s，1H)，5.30-5.39(m，1H)，5.53-5.62(m，1H)；MS?m/z?580(M ⁺+1)。

Step 4: [18-(tert-butyl group-dimethyl-silanyloxy base)-4-cyclopropane sulfonyl amino carbonyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl]-preparation of carbamic acid tertiary butyl ester

With 14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] (500mg 0.86mmol) is dissolved among the 25mL THF 19-7-alkene-4-carboxylic acid, and (180mg 1.12mmol) handles with CDI.(take every caution against error, use the glass drying oven of oven dry, avoid moistening, the N2 atmosphere keeps dry).After reaction mixture refluxed 2 hours, it is cooled to room temperature, with the cyclopropyl sulfonamide (135mg, 1.12mmol) and DBU (170mg, 1.12mmol) sequential processing.Reactant mixture stirring at room 4 hours, is removed THF through rotary evaporation.Residue is distributed between the buffer of ethyl acetate and pH value 4.Dry (MgSO ₄) organic facies, vacuum concentration obtains bullion.Use purification by flash chromatography (with 33% ethyl acetate/hexane eluting) then, obtain 300mg (51%) [18-(tert-butyl group-dimethyl-silanyloxy base)-4-cyclopropane sulfonyl amino carbonyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl]-carbamic acid tertiary butyl ester white solid. ¹H?NMR(300MHz，CD ₃OD)δ1H?0.07(s，3H)，0.08(s，3H)，0.85(s，9H)，0.87-1.49(m，21H)，1.73-1.95(m，3H)，2.08-2.16(m，1H)，2.25-2.36(m，2H)，2.42-2.56(m，1H)，2.85-2.93(m，1H)，3.65-3.74(dd，J＝10.61，3.66Hz，1H)，3.89(d，J＝10.25Hz，1H)，4.34(m，J＝9.70，9.70Hz，1H)，4.43(t，J＝7.87Hz，1H)，4.57(s，1H)，4.94-5.01(m，1H)，5.10(d，J＝8.78Hz，1H)，5.66-5.75(m，1H)，6.55(s，1H)，10.13(s，1H)；MS?m/z?683(M ⁺+1)。

Step 5: embodiment 19, (4-cyclopropane sulfonyl amino carbonyl-18-hydroxyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl)-preparation of carbamic acid tertiary butyl ester

To [18-(tert-butyl group-dimethylsilyl oxygen base)-4-cyclopropane sulfonyl amino carbonyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl]-the carbamic acid tertiary butyl ester (330mg, add in 25mL THF mixture 0.48mmol) tetrabutylammonium fluoride (150mg, 0.54mmol).Reactant mixture stirring at room 18 hours, is then removed THF through rotary evaporation.Residue is distributed between ethyl acetate and water.Dry (MgSO ₄) organic facies, vacuum concentration obtains bullion.Through grinding purification, obtain 200mg (73%) (4-cyclopropane sulfonyl amino carbonyl-18-hydroxyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 then with hexane ^4,6] 19-7-alkene-14-yl)-the carbamic acid tertiary butyl ester, embodiment 19, white solid. ¹H?NMR(500MHz，CD ₃Cl)δ1.87-1.64(m，21H)，1.70-1.98(m，3H)，2.15-2.56(m，5H)，2.85-2.94(m，1H)，3.71(d，J＝13.91Hz，1H)，4.10-4.26(m，2H)，4.51(t，J＝7.87Hz，1H)，4.62(s，1H)，4.98(m，1H)，5.06(d，J＝8.78Hz，1H)，5.64-5.71(m，1H)，6.72(s，1H)，10.24(s，1H)；MS?m/z?569(M ⁺+1)。

Use the method described in the embodiment 29 and 30, prepare the alcohol intermediate of following macro ring:

Use the method described in the embodiment 29 and 30, can prepare the alcohol intermediate of following macro ring:

Embodiment 31

The preparation of embodiment 31,2 (S)-t-butoxycarbonyl amino-3-penta-4-thiazolinyl thiohydracrylic acid

Step 1: in room temperature, (3.36g adds triethylamine (10.8mL) and 1-bromine penta-4-alkene (3.19g, 21mmol, 1.5 equivalents) in methanol 0.014mol) (166mL) solution, at room temperature the solution stirring that obtains is spent the night to N-Boc-cysteine methyl ester.Vacuum concentrated mixture with purification by flash chromatography (hexane, ethyl acetate gradient), provides 1.76g (41%) needed thioether with the residual mixture that obtains then. ¹H?NMR(500MHz，CDCl ₃)δ1.43(s，9H)，1.64(m，?2H)，2.11(m，2H)，2.51(m，2H)，2.95(m，2H)，3.75(s，3H)，4.51(m，1H)，4.95-5.03(m，2H)，5.34(m，1H)，5.80(1H，m)；MS?m/z?304(M ⁺+1)。

Step 2: (9.51g 31.4mmol) joins in the mixture of 1M LiOH/ water (200mL) and THF (200mL), at room temperature with the mixture stirred overnight that obtains with the thioether product of step 1.Use 1N hydrochloric acid with the reactant mixture acidify then, the mixture that obtains is extracted several times with ethyl acetate.The united extraction thing is used dried over mgso, and vacuum concentration provides needed acid, and embodiment 20, and it can use by ortho states in next one reaction.

Embodiment 32

Embodiment 32, the preparation of N-tertbutyloxycarbonyl-3-(4-pentenyl sulfenyl)-L-valine

The preparation of step 1:N-tertbutyloxycarbonyl-3-(4-pentenyl sulfenyl)-L-valine methyl ester

At room temperature, to 7.12g (48mmol, 1.0eq) 100mL 1 of L-3-trolovol; Add 9.60ml (96mmol in the solution of 4-diox and 25mL water; 2.0eq) the 10N sodium hydrate aqueous solution, then dropwise add 12.00mL (101mmol, 2.1eq) 5-bromo-1-amylene with a few minutes.At room temperature the mixture that obtains was stirred 68 hours.At this moment, (57mmol, 1.2eq) two dimethyl dicarbonate butyl esters at room temperature stirred the mixture 6 hours in addition to add 12.50g.Vacuum concentrated mixture, residue is soluble in water.Wash aqueous mixture with diethyl ether, use 1N hydrochloric acid to be adjusted to pH value 3, then use ethyl acetate extraction.With the extract that brine wash merges, use anhydrous magnesium sulfate drying, filter vacuum concentration.

(12.20g) is dissolved in the 120mL anhydrous dimethyl sulfoxide with bullion.In this solution, add 10.50g (76mmol) potassium carbonate and 4.70mL (76mmol) iodomethane, at room temperature the mixture that obtains was stirred 24 hours.The dilute with water reactant mixture is used ethyl acetate extraction.The extract that water (2X) and brine wash merge is used anhydrous sodium sulfate drying, filters vacuum concentration.Carry out silica gel column chromatography (eluting: the 2-10% ethyl acetate/hexane), 8.54g N-tertbutyloxycarbonyl-3-(4-pentenyl sulfenyl)-L-valine methyl ester water white oil is provided.NMR (300MHz, CDCl ₃): δ 5.76 (d of d of t, 1H, J=17.2,10.3,6.6Hz), 5.35 (br d, 1H, J=9.0Hz); 5.05-4.94 (m, 2H), 4.27 (br d, 1H, J=9.0Hz), 3.73 (s, 3H); 2.52 (m, 2H), 2.13 (quartet, 2H, J=7.3Hz), 1.61 (quintet, 2H; J=7.3Hz), 1.43 (s, 9H), 1.35 (s, 3H), 1.33 (s, 3H).

Step 2: embodiment 32, the preparation of N-tertbutyloxycarbonyl-3-(4-pentenyl sulfenyl)-L-valine

At room temperature, the 50mL aqueous solution that in the 200mL tetrahydrofuran solution of 8.52g (25.7mmol) N-tertbutyloxycarbonyl-3-(4-pentenyl sulfenyl)-L-valine methyl ester, adds 1.10g (26.2mmol) lithium hydroxide monohydrate.At room temperature the mixture that obtains was stirred 65 hours.In reactant mixture, add 28mL 1.00N hydrochloric acid then.With diethyl ether diluted mixture thing, water (3X) and brine wash are used anhydrous sodium sulfate drying, filter, and vacuum concentration obtains 8.10gN-tertbutyloxycarbonyl-3-(4-pentenyl sulfenyl)-L-valine water white oil.NMR (300MHz, CDCl ₃): δ 5.75 (d of d of t, 1H, J=17.2,10.3,6.6Hz), 5.40 (br s, 1H); 5.05-4.94 (m, 2H), 4.28 (br s, 1H), 2.56 (m, 2H); 2.13 (quartet, 2H, J=7.3Hz), 1.63 (quintet, 2H, J=7.3Hz); 1.44 (s, 9H), 1.39 (s, 3H), 1.37 (s, 3H).

Embodiment 33

Embodiment 33, the preparation of 5-allyloxy-2 (S)-(t-butoxycarbonyl amino) valeric acid

Step 1: the preparation of pyrrolidine-5-ketone-2 (S)-carboxylic acid isopropyl esters

Dean-Rodney Stark couch the separator that use to change (through be full of 4 Soxhlet extractor of molecular sieve carries out condensate and refluxes); In nitrogen atmosphere; With L-pyroglutamic acid (Aldrich; 25.0g; 195mmol) (3.71g, solution 19.5mmol) refluxed 6 hours in isopropyl alcohol (40mL) with the p-methyl benzenesulfonic acid monohydrate.After being cooled to room temperature, use the ether diluting reaction, with saturated sodium bicarbonate aqueous solution, saturated NaCl solution washing then, dry (MgSO ₄), evaporation obtains colourless serosity.Place post crystallization.In hexane, grind the crystal residue, 31.9g (96%) pyrrolidine-5-ketone-2 (S)-carboxylic acid isopropyl esters white prism be provided: ¹H NMR (300MHz, the δ 6.35 of chloroform-D) (br s, 1H), 5.04 (sept..1H, J=6.2Hz), 4.18 (dd, 1H, J=8.4,5.3Hz), 2.51-2.28 (m, 3H), 2.27-2.12 (m, 1H), 1.24 (d, 6H, J=6.2Hz) ..LCMS m/z 172 (M+H) ⁺

The preparation of step 2:1-(tertbutyloxycarbonyl)-pyrrolidine-5-ketone-2 (S)-carboxylic acid isopropyl esters

At room temperature, at N ₂In the atmosphere, with pyrrolidine-5-ketone-2 (S)-carboxylic acid isopropyl esters (product of step 26A, 31.9g, 188mmol), two dimethyl dicarbonate butyl esters (48.6g, 225mmol) and DMAP (2.30g, acetonitrile 8.8mmol) (300mL) solution stirring 30 minutes.With reactive evaporation to about 100mL, with the ether dilution, with 1N HCl, saturated NaCl solution washing then, drying (MgSO ₄), evaporation obtains 1-(tertbutyloxycarbonyl) pyrrolidine-5-ketone-2 (S) carboxylic acid isopropyl esters light yellow oil, 50.1g (99%): ¹H NMR (300MHz, and the δ 5.06 of chloroform-D) (sept.1H, J=6.2Hz), 4.53 (dd, 1H, J=9.5; 2.9Hz), 2.66-2.40 (m, 2H), 2.36-2.22 (m, 1H); 2.03-1.93 (m, 1H), 1.47 (s, 9H), 1.26 (d; 3H, J=6.2Hz), 1.24 (d, 3H, J=6.2Hz) ..LCMS m/z 272 (M+H) ⁺

The preparation of step 3:2 (S)-(t-butoxycarbonyl amino)-5-hydroxypentanoic acid isopropyl esters

With 1.5 hours to 1-(tertbutyloxycarbonyl) pyrrolidine-5-ketone-2 (S)-carboxylic acid isopropyl esters (product of step 26B, 49.5g, the sodium borohydride of adding～1g share in methanol 183mmol) (300mL) solution (10.0g, 263mmol).Other stirring reaction is 10 minutes in nitrogen atmosphere.Dilute with water extracts with ether, with the organic fraction that saturated NaCl solution washing merges, and dry (MgSO ₄), evaporation obtains light yellow oil.Carry out flash chromatography (silica gel, 20-30% ethyl acetate/hexane), obtain 31.8g (64%) 2 (S)-colourless serosity of (t-butoxycarbonyl amino)-5-hydroxypentanoic acid isopropyl esters: ¹H NMR (300MHz, the δ 5.16 of chloroform-D) (br d, 1H, J=7.3Hz), 5.03 (sept., 1H, J=6.2Hz); 4.28 (br d, 1H, J=6.2Hz), 3.67 (br dd, J=10.2,5.5Hz), 1.94-1.79 (m; 2H), 1.76-1.67 (m, 1H), 1.66-1.56 (m, 2H), 1.43 (s, 9H); 1.25 (d, 3H, J=6.2Hz), 1.23 (d, 3H, J=6.2Hz) .LCMS m/z276 (M+H) ⁺

Step 4: the preparation of isopropyl-5-allyloxy-2 (S)-(t-butoxycarbonyl amino) valerate

In nitrogen atmosphere, with 2 (S)-(t-butoxycarbonyl amino)-5-hydroxypentanoic acid isopropyl esters (product of step 26C, 17.6g, 63.9mmol), the allyl methyl carbonic ester (24.0mL, 213mmol), Pd ₂(dba) ₃(1.62g, 1.78mmol) and BINAP (4.42g, THF 7.10mmol) (150mL) degassing mixture refluxed 3 hours.After being cooled to room temperature, use the ether diluting reaction, through diatomite filtration, evaporation obtains the crineous serosity.Residue is carried out flash chromatography (silica gel, 30% ether/hexane), obtains 5-allyloxy-2 (S)-heavy-gravity water white oil of (t-butoxycarbonyl amino) valeric acid isopropyl esters, 16.3g (81%): ¹H NMR (300MHz, the δ 5.88 (ddt, 1H, 17.4,10.4,5.5) of chloroform-D), 5.28 (m, 1H), 5.22-5.11 (m; 1H), 5.02 (sept., 1H, J=6.2Hz), 4.21 (br t, 1H, J=6.7Hz), 3.94 (dt, 2H; J=5.9,1.5Hz), 3.42 (t, 2H, J=5.9Hz), 1.901.82 (m, 1H), 1.75 1.57 (m, 3H); 1.42 (s, 9H), 1.21 (d, 3H, J=6.2Hz), 1.19 (d, 3H, J=6.2Hz) ..LCMS m/z 316 (M+H) ⁺

Step 5: embodiment 33, the preparation of 5-allyloxy-2 (S)-(t-butoxycarbonyl amino) valeric acid

At room temperature; In nitrogen atmosphere, with 5-allyloxy-2 (S)-(t-butoxycarbonyl amino) valeric acid isopropyl esters (product of step 26D, 16.1g; 51.1mmol) and lithium hydroxide monohydrate (4.19g, mixture 102mmol) stirred 16 hours in THF/ water (100mL/20mL).Dilute with water reaction, with the ether washing, the pH value of aqueous solution fraction is adjusted to～4, extract with ether, with the organic fraction that saturated NaCl washing merges, drying (MgSO ₄), evaporation obtains 5-allyloxy-2 (S)-(t-butoxycarbonyl amino) valeric acid pale yellow syrup: ¹H NMR (300MHz, the δ 5.89 (ddt, 1H, J=17.4,10.4,5.5) of chloroform-D), 5.25 (dd, 1H; J=17.4,1.6Hz), 5.17 (dd, 1H, J=10.4,1.6Hz), 4.30 (br d, 1H; J=6.2), 3.96 (dt, 2H, J=5.9,1.5Hz), 3.46 (t, 2H, J=5.9Hz); 1.961.86 (m, 1H), 1.851.77 (m, 1H), 1.751.64 (m, 2H), 1.43 (s, 9H) ..LCMS m/z 274 (M+H) ⁺

Embodiment 34

The conventional method of preparation embodiment 34

Be prepared as follows embodiment 23: the DMF solution of the threonine of N-trityl as protecting group is joined in the DMF solution (being cooled to-15 ℃) of sodium hydride.-15 ℃ of stirred reaction mixtures 30 minutes, then add 5-bromo-1-amylene, with the mixture heated that obtains to-5 ℃.-5 ℃ of maintenance reactant mixtures 3 days, then come the quencher reaction through adding 1N HCl aqueous solution, use aforesaid standard extraction methods to carry out post processing.Utilize the standard colour chart method, obtain the embodiment 23 of pure formula.

Embodiment 35:

Embodiment 35, the preparation of N-tertbutyloxycarbonyl-O-(4-pentenyl)-L-serine

The preparation of step 1:N-tertbutyloxycarbonyl-O-(4-pentenyl)-L-serine methyl ester

At room temperature, (50mmol 1.0eq) adds 2.00g (50mmol, 1.0eq) 60% sodium hydride/mineral oil in the 500mL anhydrous dimethyl sulfoxide solution of N-tertbutyloxycarbonyl-L-serine to 10.26g.At room temperature stirred this mixture 0.5 hour, till stopping to emit gas.(50mmol, 1.0eq) 5-bromo-1-amylene then add other 2.00g (50mmol, 1.0eq) 60% sodium hydride/mineral oil immediately in the solution that obtains, to add 6.00mL.Then reactant mixture was at room temperature stirred 16 hours.With 2000mL water diluted mixture thing, through adding 50mL 1.00N hydrochloric acid pH value is adjusted to 3-4, use ethyl acetate extraction.Water (2X) and brine wash organic facies are used anhydrous sodium sulfate drying, filter vacuum concentration.Remove relict mineral oil, the material that obtains is dissolved in the dilute sodium hydroxide aqueous solution.Use the hexane wash aqueous solution, then use hydrochloric acid to be adjusted to pH value 4, use ethyl acetate extraction.Water (2X) and brine wash extract are used anhydrous sodium sulfate drying, filter vacuum concentration.

(7.70g) is dissolved in the 100mL anhydrous dimethyl sulfoxide with bullion.In this solution, add 7.80g (56mmol) potassium carbonate and 3.50mL (56mmol) iodomethane, at room temperature the mixture that obtains was stirred 24 hours.The dilute with water reactant mixture is used ethyl acetate extraction.The extract that water (2X) and brine wash merge is used anhydrous sodium sulfate drying, filters vacuum concentration.Carry out silica gel column chromatography (eluting: the 2-10% ethyl acetate/hexane), 6.70g N-tertbutyloxycarbonyl-O-(4-pentenyl)-L-serine methyl ester water white oil is provided.NMR (300MHz, CDCl ₃): δ 5.78 (dof d of t, 1H, J=17.2,10.2,6.6Hz), 5.34 (br d, 1H, J=8.0Hz); 5.03-4.92 (m, 2H), 4.40 (m, 1H), 3.81 (d of d, 1H, J=9.5,2.9Hz); 3.74 (s, 3H), 3.61 (d ofd, 1H, J=9.5,3.5Hz), 3.42 (m, 2H); 2.06 (quartet, 2H, J=7.3Hz), 1.61 (quintet., 2H, J=7.3Hz), 1.44 (s, 9H).

Step 2: embodiment 35, the preparation of N-tertbutyloxycarbonyl-O-(4-pentenyl)-L-serine

At room temperature, the 100mL aqueous solution that in the 500mL tetrahydrofuran solution of 6.65g (23mmol) N-tertbutyloxycarbonyl-O-(4-pentenyl)-L-serine methyl ester, adds 1.95g (46mmol) lithium hydroxide monohydrate.At room temperature the mixture that obtains was stirred 40 hours.In reactant mixture, add 46mL 1.00N hydrochloric acid then.With ethyl acetate diluted mixture thing, water (3X) and brine wash are used anhydrous sodium sulfate drying, filter, and vacuum concentration obtains 6.30g N-tertbutyloxycarbonyl-O-(4-pentenyl)-L-serine water white oil...NMR (300MHz, CDCl ₃): δ 5.77 (dof d of t, 1H, J=17.2,10.2,6.6Hz), 5.37 (br d, 1H, J=8.0Hz); 5.03-4.92 (m, 2H), 4.42 (m, 1H), 3.87 (d of d, 1H, J=9.5,2.6Hz); 3.63 (d of d, 1H, J=9.5,4.0Hz), 3.45 (t, 2H, J=6.6Hz), 2.07 (quartets; 2H, J=7.3Hz), 1.64 (quintet, 2H, J=7.3Hz), 1.44 (s, 9H).

Embodiment 36

Embodiment 36, (S)-and the butyro-preparation of 4-allyloxy-2-(t-butoxycarbonyl amino)

At 0 ℃, to sodium hydride (913mg, add in DMF mixture 22.8mmol) the N-t-Boc-L-homoserine (2g, 9.13mmol).Stirred this reactant mixture 15 minutes at 0 ℃, then add allyl bromide, bromoallylene (1.38g, 11.4mmol).Mixture is warming up to room temperature, stirred 2 hours.Then with its vacuum concentration.The dilute with water residue is with hexane and the washing of ether order.Remove organic layer, water layer is adjusted to pH value 3 carefully with 1N HCl.With this acidic aqueous solution of ethyl acetate extraction.Dry (MgSO ₄) organic facies, vacuum concentration, obtain 2.2g (93%) (S)-4-allyloxy-2-(t-butoxycarbonyl amino) butanoic acid water white oil. ¹H?NMR(300MHz，CD ₃OD)δ1.42(s，9H)，1.80-1.90(m，1H)，2.04-2.16(m，1H)，3.50-3.54(m，2H)，3.97(d，J＝4.39Hz，2H)，4.23(dd，J＝8.78，4.39Hz，1H)，5.15(d，J＝10.25Hz，1H)，5.26(dd，J＝17.38，1.65Hz，1H)，5.84-5.97(m，1H)。

Embodiment 37

Embodiment 37, (S)-and the preparation of 2-(tertbutyloxycarbonyl)-3-(2-nitro-N-(penta-4-thiazolinyl) phenyl sulfonamido) propanoic acid

The preparation of amino-2 (S) of step 1:3--(t-butoxycarbonyl amino) methyl propionate

In the 50mL dichloromethane mixture of i (Boc-DAP-OH) (3.0g 14.7mmol), add 5mL methanol.In this solution, add at leisure (trimethyl silyl) Azimethylene. (2M, in ether, 7.9mL, 15.8mmol).In stirring at room mixture 2 hours, till all solids dissolving, this solution became light yellow.Then it is concentrated, obtain amino-2 (S) of 3.2g (99%) 3--(t-butoxycarbonyl amino) methyl propionate ii water white oil. ¹H?NMR(CD ₃OD，300MHz)δ?1.46(s，9H)，2.82-3.00(m，2H)，3.71(s，3H)，4.14(brs，1H)。

The preparation of 2 (S)-(t-butoxycarbonyl amino)-3-(2-nitrobenzophenone sulfonamido) methyl propionate iii:

To amino-2 (S) of 3--(t-butoxycarbonyl amino) methyl propionate ii (1.6g, add in DCM 7.3mmol) (50mL) mixture DIPEA (1.64mL, 9.4mmol) with the 2-nitrobenzene sulfonyl chloride (1.62g, 7.3mmol).Mixture was at room temperature stirred 2 hours.Then it is concentrated, is dissolved in the ethyl acetate, then with it with saturated sodium bicarbonate, brine wash, use dried over mgso.With its filtration, concentrate then, obtain the yellow foam of 2.9g (98%) 2 (S)-(t-butoxycarbonyl amino)-3-(2-nitrobenzophenone sulfonamido) methyl propionate iii. ¹H?NMR(CD ₃OD，300MHz)δ1.41(s，9H)，3.36-3.51(m，2H)，3.71(s，3H)，4.22(m，1H)，7.80-7.90(m，3H)，8.07-8.10(m，1H)。

The preparation of 2 (S)-(t-butoxycarbonyl amino)-3-(2-nitro-N-(penta-4-thiazolinyl) phenyl sulfonamido) methyl propionate iv:

To 2 (S)-(t-butoxycarbonyl amino)-3-(2-nitrobenzophenone sulfonamido) methyl propionate iii (150mg, add in the mixture of 3mL DMF 0.37mmol) potassium carbonate (102mg, 0.74mmol).In this mixture of stirring at room 20 minutes, then add 5-bromo-1-amylene (65 μ L, 0.55mmol).Reactant mixture was at room temperature stirred 2 days.Filter then, concentrate,, obtain 75mg (43%) 2 (S)-(tert-butoxycarbonyl is amino)-3-(2-nitro-N-(penta-4-thiazolinyl) phenyl sulfonamido) methyl propionate iv yellow solid through silica gel chromatography purification (with 25% ethyl acetate/hexane eluting). ¹H?NMR(CD ₃OD，300MHz)δ1.42(s，9H)，1.54-1.64(m，2H)，1.97(q，J＝7.20Hz，2H)，3.37(m，2H)，3.57-3.80(m，2H)，3.72(s，3H)，4.42(dd，J＝8.60，5.31Hz，1H)，4.91-5.01(m，2H)，5.69-5.79(m，1H)，7.75-7.85(m，3H)，8.04(m，1H)；MS?m/z?372(M ⁺+1-Boc)。

The preparation of embodiment 37,2 (S)-(t-butoxycarbonyl amino)-3-(2-nitro-N-(penta-4-thiazolinyl) phenyl sulfonamido) propanoic acid v:

(500mg 1.06mmol) is dissolved in the following mixed solvent system: THF (4ml), methanol (1mL) and water (2mL) with (S)-2-(tertbutyloxycarbonyl)-3-(2-nitro-N-(penta-4-thiazolinyl) phenyl sulfonamido)-propanoic acid methyl ester iv.Adding powder Lithium hydrate (250mg, 10.4mmol).This pale yellow syrup of stirring at room 15 hours, vacuum concentration then.Residue is distributed between ether and water.Remove the ether phase, handle water with 1N HCl, till pH value 4.With this acid solution of ethyl acetate extraction four times.With the dry (MgSO of the ethyl acetate extract that merges ₄), vacuum concentration obtains 430mg (89%) 2-(t-butoxycarbonyl amino)-3-(2-nitro-N-(penta-4-thiazolinyl) phenyl sulfonamido) propanoic acid (embodiment 26) yellow oil. ¹H?NMR(CD ₃OD，300MHz)δ1.38(s，9H)，1.51-1.60(m，2H)，1.89-1.98(m，2H)，3.28-3.32(m，2H)，3.59-3.64(dd，J＝14.95，9.46Hz，1H)，3.71-3.74(m，1H)，4.33(dd，J＝9.61，4.43Hz，1H)，4.87-4.94(m，2H)，5.63-5.72(m，1H)，7.71-7.77(m，3H)，8.01(dd，J＝7.48，1.37Hz，1H)；MS?m/z?358(M ⁺+1-Boc)。

Embodiment 86

(1S, 4R, 6S, 14S, 18R)-[7-cis-4-cyclopropane sulfonyl amino carbonyl-12-cyclopropyl-18-hydroxyl-2,15-dioxo-3,12,16-three azepines-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl] preparation of carbamic acid tertiary butyl ester (embodiment 86)

Synthesizing of step 1:N-(penta-4-thiazolinyl) cyclopropylamine.

Use to add funnel, with 5 minutes time, (15.75g, 50mL methanol solution 106mmol) join cyclopropylamine, and (20.6g was in 200mL methanol solution 361mmol) with 5-bromine amylene.This solution of stirring at room 72 hours, refluxed 1 hour this moment.Distillation for removing methanol and excessive cyclopropylamine.Residue (hydrobromate of product) is distributed between ether and 4N NaOH.With ether (2x) washing water.With the dry (MgSO of the ether extract that merges ₄), filter, concentrate, obtain 8g (60%) N-(penta-4-thiazolinyl) cyclopropylamine yellow oil: ¹H NMR (500MHz, CDCl ₃) δ 0.31-0.36 (and m, 2H) 0.40-0.46 (m, 2H) 1.53-1.63 (m, 2H) 1.87 (brs, 1H) 2.05-2.10 (m, 2H) 2.10-2.14 (m, 1H) 2.69 (t, J=7.32Hz, 2H) 4.91-5.07 (m, 2H) 5.72-5.88 (m, 1H).

Synthesizing of step 2:2 (S)-(t-butoxycarbonyl amino)-3-[N-cyclopropyl-N-(penta-4-thiazolinyl) amino] propanoic acid

(668mg, 20mL acetonitrile mixture 5.30mmol) join N-tertbutyloxycarbonyl-L-serine β-lactone, and (1.0g is in 40mL acetonitrile serosity 5.30mmol) with N-(penta-4-thiazolinyl) cyclopropylamine.At N ₂In the atmosphere, in this mixture of stirring at room 5 days, then vacuum concentration obtains bullion 2 (S)-(t-butoxycarbonyl amino)-3-[N-cyclopropyl-N-(penta-4-thiazolinyl) amino] propanoic acid.This material just can use in step 3 without purification.LC-MS (Phenomenex 10 micromoles (" μ m ") C18HPLC post: 3.0 * 50mm length.Gradient: 100% solvent orange 2 A/0% solvent B to 0% solvent orange 2 A/100% solvent B.The gradient time: 3min.The time of staying: 1min.Flow velocity: 4mL/min.Detector wavelength: 220nM.Solvent orange 2 A: 10%MeOH/90%H ₂O/0.1%TFA.Solvent B:10%H ₂O/90%MeOH/0.1%TFA.(retention time: 2.50min), MS m/z 313 (M ⁺+ 1).

Synthesizing of step 3:1 (R)-[1-[2 (S)-(t-butoxycarbonyl amino)-3-[N-cyclopropyl-N-(penta-4-thiazolinyl) amino] propiono]-4 (R)-hydroxyl pyrrolidine-2 (S)-carboxylic acid amides]-2 (S)-vinyl cyclopropane carboxylic acid acetoacetic esters.

With 2 (S)-(t-butoxycarbonyl amino)-3-[N-cyclopropyl-N-(penta-4-thiazolinyl) amino] propanoic acid (1.47g; 4.71mmol) 20mL DCM solution with 1 (R)-[4 (R)-hydroxyl pyrrolidine-2 (S)-carboxylic acid amides]-2 (S)-vinyl cyclopropane-carboxylic acid carbethoxy hydrochlorides (in embodiment 5 preparation) (1.44g; 4.71mmol), N-methylmorpholine (1.80mL; 16.34mmol) and HATU (2.14g, 5.53mmol) sequential processing.In room temperature, at N ₂Stirred reaction mixture is 3 hours in the atmosphere, then vacuum concentration.Residue is soluble in water, add 1N HCl, till pH value=5.Extract aqueous solution with EtOAc (3x).Use saturated NaHCO ₃The solution washing organic facies, dry (MgSO ₄), vacuum concentration obtains bullion.Carry out flash chromatography (50% ethyl acetate/hexane to 100% ethyl acetate), obtain 1.55g (58%) 1 (R)-[1-[2 (S)-(t-butoxycarbonyl amino)-3-[N-cyclopropyl-N-(penta-4-thiazolinyl) amino] propiono]-4 (R)-hydroxyl pyrrolidine-2 (S)-carboxylic acid amides]-2 (S)-vinyl cyclopropane carboxylic acid acetoacetic ester white foams:

LC-MS (Phenomenex-Luna S10 HPLC post: 3.0 * 50mm length.Gradient: 100% solvent orange 2 A/0% solvent B to 0% solvent orange 2 A/100% solvent B.The gradient time: 2min.The time of staying: 1min.Flow velocity: 4mL/min.Detector wavelength: 220nM.Solvent orange 2 A: 10%MeOH/90%H ₂O/0.1%TFA.Solvent B:10%H ₂O/90%MeOH/0.1%TFA).(retention time: 1.38min), MS m/z 564 (M ⁺+ 1).

Synthesizing of step 4:1 (R)-[1-[2 (S)-(t-butoxycarbonyl amino)-3-[N-cyclopropyl-N-(penta-4-thiazolinyl) amino] propiono]-4 (R)-(the silica-based oxygen base of tert-butyl group dimethyl) pyrrolidine-2 (S)-carboxylic acid amides]-2 (S)-vinyl cyclopropane carboxylic acid acetoacetic esters.

To 1 (R)-[1-[2 (S)-(t-butoxycarbonyl amino)-3-[N-cyclopropyl-N-(penta-4-thiazolinyl) amino] propiono]-4 (R)-hydroxyl pyrrolidine-2 (S)-carboxylic acid amides]-2 (S)-vinyl cyclopropane carboxylic acid acetoacetic ester (1.55g; 2.75mmol) the mixture of 10mL DMF in add imidazoles (0.47g; 6.88mmol) and tert-butyldimethylsilyl chloride (826mg, 5.50mmol).At this mixture of stirring at room 18 hours, vacuum concentration, and between ethyl acetate and water, distribute.Use the dried over mgso organic facies, vacuum concentration obtains the off-white color solid.Carry out flash chromatography (with dichloromethane eluent ethyl acetate then); Obtain 1 (R)-[1-[2 (S)-(t-butoxycarbonyl amino)-3-[N-cyclopropyl-N-(penta-4-thiazolinyl) amino] propiono]-4 (R)-(the silica-based oxygen base of tert-butyl group dimethyl) pyrrolidine-2 (S)-carboxamide groups]-2 (S)-vinyl cyclopropane carboxylic acid acetoacetic ester white solids (1.75g, 94%):

LC-MS (Phenomenex 10 μ m C18 HPLC posts: 3.0 * 50mm length.Gradient: 100% solvent orange 2 A/0% solvent B to 0% solvent orange 2 A/100% solvent B.The gradient time: 2min.The time of staying: 1min.Flow velocity: 5mL/min.Detector wavelength: 220nM.Solvent orange 2 A: 10%MeOH/90%H ₂O/0.1%TFA.Solvent B:10%H ₂O/90%MeOH/0.1%TFA).(retention time: 2.51min), MS m/z 677 (M ⁺+ 1).

Step 5: (1S, 4R, 6S, 14S, 18R)-and 7-cis-14-t-butoxycarbonyl amino-18-(the silica-based oxygen base of tert-butyl group dimethyl)-2,15-dioxo-3,12,16-three aza-tricycle [14.3.0.0 ^4,6] 19-7-alkene-4-carboxylic acid, ethyl ester synthetic.

To 1 (R)-[1-[2 (S)-(t-butoxycarbonyl amino)-3-[N-cyclopropyl-N-(penta-4-thiazolinyl) amino] propiono]-4 (R)-(the silica-based oxygen base of tert-butyl group dimethyl) pyrrolidine-2 (S)-carboxamide groups]-2 (S)-vinyl cyclopropane carboxylic acid acetoacetic ester (1.45g; 2.14mmol) the 1L dichloromethane solution in add 181mg (0.21mmol) Grubb ' s second filial generation catalyst [(1; 3-two-(2; 4, the 6-trimethylphenyl)-2-imidazolidine subunit) dichloro (phenylmethylene) (tricyclohexyl phosphine) ruthenium].With mixture reflux 1 hour.The catalyst of adding second portion (50mg, 0.058mmol), in the stirring at room mixture overnight.The vacuum concentration residue then through purification by flash chromatography, with 50% ether/hexane eluting, obtains 0.84g (62%) (1S; 4R, 6S, 14S, 18R)-7-cis-14-t-butoxycarbonyl amino-18-(the silica-based oxygen base of tert-butyl group dimethyl)-2; 15-dioxo-3,12,16-three aza-tricycle [14.3.0.0 ^4,6] 19-7-alkene-4-carboxylic acid, ethyl ester white solid: LC-MS (Phenomenex 10 μ m C18 HPLC posts: 3.0 * 50mm length.Gradient: 100% solvent orange 2 A/0% solvent B to 0% solvent orange 2 A/100% solvent B.The gradient time: 2min.The time of staying: 1min.Flow velocity: 5mL/min.Detector wavelength: 220nM.Solvent orange 2 A: 10%MeOH/90%H ₂O/0.1%TFA.Solvent B:10%H ₂O/90%MeOH/0.1%TFA).(retention time: 2.43min), MS m/z 649 (M ⁺+ 1).

Step 6: (1S, 4R, 6S, 14S, 18R)-and 7-cis-14-t-butoxycarbonyl amino-18-(the silica-based oxygen base of tert-butyl group dimethyl)-2,15-dioxo-3,12,16-three aza-tricycle [14.3.0.0 ^4,6] 19-7-alkene-4-carboxylic acid synthetic.

To (1S, 4R, 6S, 14S, 18R)-and 7-cis-14-t-butoxycarbonyl amino-18-(the silica-based oxygen base of tert-butyl group dimethyl)-2,15-dioxo-3,12,16-three aza-tricycle [14.3.0.0 ^4,6] 19-7-alkene-4-carboxylic acid, ethyl ester (0.84g, and adding powder lithium hydroxide monohydrate in THF 1.30mmol) (30mL), methanol (15mL) and water (4mL) solution (0.31g, 12.90mmol).In room temperature, at N ₂In the atmosphere with the pale yellow syrup stirred overnight that obtains.Vacuum concentrated mixture then, and between hexane/ether (1: 1) and water, distribute.Remove organic facies, handle water with 1N HCl, till pH value 5.Extract this acid solution (three times) with EtOAc.Dry (MgSO ₄) the EtOAc extract that merges, vacuum concentration, obtain 0.495g (61%) (1S, 4R, 6S, 14S, 18R)-7-cis-14-t-butoxycarbonyl amino-18-(the silica-based oxygen base of tert-butyl group dimethyl)-2,15-dioxo-3,12,16-three aza-tricycle [14.3.0.0 ^4,6] 19-7-alkene-4-carboxylic acids white solid: LC-MS (Phenomenex 10 μ mC18HPLC posts: 3.0 * 50mm length.Gradient: 100% solvent orange 2 A/0% solvent B to 0% solvent orange 2 A/100% solvent B.The gradient time: 2min.The time of staying: 1min.Flow velocity: 5mL/min.Detector wavelength: 220nM.Solvent orange 2 A: 10%MeOH/90%H ₂O/0.1%TFA.Solvent B:10%H ₂O/90%MeOH/0.1%TFA).(retention time: 2.36min), MS m/z621 (M ⁺+ 1).

Step 7: (1S, 4R, 6S, 14S, 18R)-[7-cis-4-cyclopropane sulfonyl amino carbonyl-12-cyclopropyl-18-(the silica-based oxygen base of tert-butyl group dimethyl)-2,15-dioxo-3,12,16-three azepines-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl] carbamic acid tertiary butyl ester synthetic.

Will (1S, 4R, 6S, 14S, 18R)-and 7-cis-14-t-butoxycarbonyl amino-18-(the silica-based oxygen base of tert-butyl group dimethyl)-2,15-dioxo-3,12,16-three aza-tricycle [14.3.0.0 ^4,6] (490mg 0.79mmo1) is dissolved among the 15mL THF 19-7-alkene-4-carboxylic acid, and (179mg 1.10mmol) handles with CDI.(take every caution against error, use the glass drying oven of oven dry, avoid moistening, N keeps dry ₂Atmosphere).After reaction mixture refluxed two hours, it is cooled to room temperature, with the cyclopropyl sulfonamide (134mg, 1.10mmol) and DBU (168mg, 1.10mmol) sequential processing.After stirred overnight at room temperature, remove THF through rotary evaporation.Residue is soluble in water, add 1N HCl, till pH value=5.Extract aqueous solution with EtOAc (3x).With the dry (MgSO of the EtOAc extract that merges ₄), vacuum concentration obtains bullion.Through quick column purification,, obtain 300mg (53%) (1S, 4R with 3% ethanol/methylene eluting; 6S, 14S, 18R)-[7-cis-4-cyclopropane sulfonyl amino carbonyl-12-cyclopropyl-18-(the silica-based oxygen base of tert-butyl group dimethyl)-2; 15-dioxo-3,12,16-three azepines-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl] carbamic acid tertiary butyl ester white solid: LC-MS (Phenomenex 10 μ m C18HPLC posts: 3.0 * 50mm length.Gradient: 100% solvent orange 2 A/0% solvent B to 0% solvent orange 2 A/100% solvent B.The gradient time: 2min.The time of staying: 1min.Flow velocity: 5mL/min.Detector wavelength: 220nM.Solvent orange 2 A: 10%MeOH/90%H ₂O/0.1%TFA.Solvent B:10%H ₂O/90%MeOH/0.1%TFA).(retention time: 2.40min), MS m/z 724 (M ⁺+ 1).

Step 8: (1S, 4R, 6S, 14S, 18R)-[7-cis-4-cyclopropane sulfonyl amino carbonyl-12-cyclopropyl-18-hydroxyl-2,15-dioxo-3,12,16-three azepines-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl] carbamic acid tertiary butyl ester (embodiment 86) synthetic.

To chemical compound (1S, 4R, 6S, 14S, 18R)-[7-cis-4-cyclopropane sulfonyl amino carbonyl-12-cyclopropyl-18-(the silica-based oxygen base of tert-butyl group dimethyl)-2,15-dioxo-3,12,16-three azepines-three rings [14.3.0.0 ^4,6] 19-7-alkene-14-yl] the carbamic acid tertiary butyl ester (250mg, add in the mixture of 15mL THF 0.35mmol) tetrabutyl ammonium fluoride (129mg, 0.46mmol).Mixture was at room temperature stirred 18 hours.Rotary evaporation is removed THF, between ethyl acetate and water, distributes residue.Dry (MgSO ₄) organic facies, vacuum concentration obtains bullion.Grind with hexane and to carry out purification, provide 200mg (94%) (1S, 4R, 6S, 14S, 18R)-[7-cis-4-cyclopropane sulfonyl amino carbonyl-12-cyclopropyl-18-hydroxyl-2,15-dioxo-3,12,16-three azepines-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl] carbamic acid tertiary butyl ester white solid: LC-MS (retention time: 2.32min), MS m/z 610 (M ⁺+ 1).

(Phenomenex 10 μ m C18HPLC posts: 3.0 * 50mm length.Gradient: 100% solvent orange 2 A/0% solvent B to 0% solvent orange 2 A/100% solvent B.The gradient time: 3min.The time of staying: 1min.Flow velocity: 4mL/min.Detector wavelength: 220nM.Solvent orange 2 A: 10%MeOH/90%H ₂O/0.1%TFA.Solvent B:10%H ₂O/90%MeOH/0.1%TFA).(retention time: 2.32min), MS m/z 610 (M ⁺+ 1).

Embodiment 87: the preparation of chemical compound 1

Scheme 1

Step 1:2-((1R, 2S)-1-(cyclopropyl sulfuryl amino formoxyl)-2-vinyl cyclopropyl carbamoyl)-4-Phenylpyrrolidine-1-carboxylic acid (2S, 4S)-preparation of tertiary butyl ester

To (2S; 4S)-(0.711g 2 for Boc-4-Phenylpyrrolidine-2-carboxylic acid; 44mmol) (0.948g is 7.33mmol) with cyclopropane sulfonic acid (1-(R)-amino-2-(S)-vinyl-cyclopropane carbonyl) amide HCl salt (0.561g for (from Chem-Impex International, Inc buys), diisopropylethylamine; 2.44mmol) dichloromethane (24ml) mixture in add HATU (1.21g, 3.18mmol).After the stirred reaction mixture 14 hours, it is used 5%NaHCO ₃Aqueous solution (50mL) and 5% aqueous solution of citric acid (50mL) washing.Extract each water layer in proper order with 2 * 25mL dichloromethane.Use MgSO ₄The dry organic layer that merges filters, and concentrates.The brown viscous oil that obtains is passed through flash column chromatography purification (SiO ₂, 97: 3, dichloromethane: methanol); Obtain 2-((1R; 2S)-1-(cyclopropyl sulfuryl amino formoxyl)-2-vinyl cyclopropyl carbamoyl)-4-Phenylpyrrolidine-1-carboxylic acid (2S, 4S)-the foamed solid of tertiary butyl ester brown (0.973g, 79% productive rate). ¹H?NMR(CD ₃OD，500MHz)δ0.91(br?s，1H)，1.03(d，J＝6.4Hz，1H)，1.16-1.23(m，1H)，1.31-1.38(m，1H)，1.44(dd，J＝9.6，5.6Hz，1H)，1.48(s，4H)，1.52(s，5H)，1.89(t，J＝6.4Hz)，2.09(q，J＝8.4Hz，0.4H)，2.19(q，J＝8.6Hz，0.6H)，2.85-2.90(m，0.4H)，2.92-2.97(m，0.6H)，3.44(t，J＝9.6Hz，1H)，3.65(p，J＝8.0Hz，1H)，3.93-4.01(m，1H)，4.29(dd，J＝10.1，6.7Hz，1H)，5.12(d，J＝10.1Hz，1H)，5.30(d，J＝18.0Hz，1H)，5.75-5.82(m，1H)，7.24(t，J＝6.9Hz，1H)，7.29-7.33(m，5H)；MS?m/z?504(M+Na)。

Step 2: (2S, 4S)-N-(preparation of (1R, 2S)-1-(cyclopropyl sulfuryl amino formoxyl)-2-vinyl cyclopropyl)-4-Phenylpyrrolidine-2-carboxylic acid amides

(0.900g adds TFA (5mL) in dichloromethane 1.79mmol) (5mL) solution to the product of the step 1 of embodiment 1.After at room temperature stirring 30 minutes, concentrated reaction mixture, vacuum drying slightly.The brown viscous oil that obtains is dissolved in the dichloromethane (10mL) again, and dropwise joins in 1N HCl/ diethyl ether (10mL) solution.Formation white solid deposition is collected through vacuum filtration.Use the diethyl ether washing leaching cake, obtain 0.720mg (91% productive rate) (2S, 4S)-N-((1R, 2S)-1-(cyclopropyl sulfuryl amino formoxyl)-2-vinyl cyclopropyl)-4-Phenylpyrrolidine-2-carboxylic acid amides.MS?m/z?404(MH ⁺)。

Step 3: (S)-1-((2S, 4S)-2-((1R, 2S)-1-(cyclopropyl sulfuryl amino formoxyl)-2-vinyl cyclopropyl carbamoyl)-4-Phenylpyrrolidine-1-yl)-preparation of 1-oxo ninth of the ten Heavenly Stems-8-alkene-2-aminocarbamic acid tertiary butyl ester

To (S)-2-(t-butoxycarbonyl amino) ninth of the ten Heavenly Stems-8-olefin(e) acid (buying) from RSP Amino Acids (140mg, add in 20mL DMF solution 0.516mmol) step 2 of embodiment 1 product (227mg, 0.516mmol).In this solution, add HATU (236mg, 0.620mmol), then add N-methylmorpholine (182mg, 1.80mmol).In this mixture of stirring at room 4 hours, then vacuum concentration was removed DMF.Residue is being distributed (keeping water between pH value 3 to pH value 5) between 0.1N HCl and the ethyl acetate.Dry (MgSO ₄) organic facies, filter, concentrate, obtain 300mg bullion brown oil.Flash chromatography is with 1-5% methanol/CH ₂Cl ₂Eluting; Obtain 230mg (68%) (S)-1-((2S; 4S)-2-((1R, 2S)-1-(cyclopropyl sulfuryl amino formoxyl)-2-vinyl cyclopropyl carbamoyl)-4-Phenylpyrrolidine-1-yl)-1-oxo ninth of the ten Heavenly Stems-8-alkene-2-aminocarbamic acid tertiary butyl ester water white oil.MS?m/z?657(MH ⁺)。

Step 4: the preparation of chemical compound 1

(124mg 0.189mmol) is dissolved in CH with the product of step 3 ₂Cl ₂In, be placed on N ₂In the atmosphere, (16mg 0.019mmol) handles with Grubb ' s second filial generation RCM catalyst (tricyclohexyl phosphine [1,3-two (2,4,6-trimethyl-phenyl)-4,5-glyoxalidine-2-subunit] [benzal]-ruthenous chloride (IV)).After refluxing 18 hours, orange/red solution is cooled to room temperature, vacuum concentration obtains the 125mg brown oil.Flash chromatography is with 1-5% methanol/CH ₂Cl ₂Eluting, (～60% purity is utilized to obtain 60mg bullion macro ring product ¹H NMR and LC/MS assessment).Utilize preparation HPLC, obtain 22mg pure products off-white color solid. ¹H?NMR(500MHz，CDCl ₃)δppm0.90-1.00(m，1H)，1.05-1.14(m，1H)，1.14-1.22(m，1H)，1.23-1.84(m，10H)，1.41(s，9H)，1.88-2.00(m，2H)，2.14-2.29(m，2H)，2.39-2.49(m，1H)，2.50-2.60(m，1H)，2.86-2.96(m，1H)，3.91-4.01(m，2H)，4.05-4.14(m，1H)，4.39-4.48(m，1H)，4.48-4.56(m，1H)，4.97-5.04(m，1H)，5.13(d，J＝8.24Hz，1H)，5.70(q，J＝8.95Hz，1H)，6.72(s，1H)，7.36-7.24(m，5H)，10.14(s，1H)..MS?m/z?629(MH ⁺)。

Embodiment 88: the preparation of chemical compound 2

With 2 (S)-t-butoxycarbonyl amino-8-nonenoic acid (buying) (3.5g from RSP Amino Acids; 12.9mmol) the 200mL dichloromethane solution with 4 (R)-hydroxyl pyrrolidines-2 (S)-carboxylic acid methyl ester hydrochloride (2.15g; 11.8mmol), N-methylmorpholine (4.25mL; 38.6mmol) and HATU (5.37g, 14.1mmol) sequential processing.At room temperature, at N ₂Stirred reaction mixture is 3 days in the atmosphere, then vacuum concentration.Residue is distributed between the buffer (biphthalate) of ethyl acetate and pH value 4.Use saturated NaHCO ₃The solution washing organic facies, dry (MgSO ₄), filtering, vacuum concentration obtains bullion.Carry out flash chromatography (50% ethyl acetate/hexane to 100% ethyl acetate), obtain 1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (the S)-carboxylic acid methyl ester water white oils of 4.7g (～100%). ¹H NMR (500MHz, CD ₃OD) δ 1.33-1.50 (m, 8H), 1.46 (s, 9H), 1.57 (m, 1H), 1.72 (m, 1H) 2.08 (m; 2H), 2.28 (m, 1H), 3.72 (s, 3H) 3.75-3.87 (m, 2H); 4.36 (m, 1H), 4.51 (bs, 1H), 4.57 (t, J=8.2Hz, 1H); 4.95 (d, J=10.4Hz, 1H), 5.01 (m, 1H), 5.83 (m, 1H); LC-MS (method A, retention time: 3.01min), MS m/z 399 (M ⁺+ 1).

Step 2:1-{ [1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (S) carbonyls]-(1R)-amino } preparation of-2 (S)-vinyl-cyclopropane carboxylic acid acetoacetic ester.

(4.7g 11.8mmol) is dissolved in THF (80mL), methanol (20mL) and the water (40mL) with 1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (S)-carboxylic acid methyl ester.Adding powder Lithium hydrate (5.6g, 233mmol).At room temperature, at N ₂Stirred pale yellow syrup 16 hours in the atmosphere, then vacuum concentration.Residue is distributed between ether and water.Remove the ether phase, handle water with 1N HCl, till pH value 4.With this acid solution of ethyl acetate extraction (three times).With the dry (MgSO of the ethyl acetate extract that merges ₄), filtering, vacuum concentration obtains 1-(2 (S)-t-butoxycarbonyl amino-8-nonene acyl group)-4 (R)-hydroxyl-pyrrolidine-2 (the S)-carboxylic acid white solids of 4.36g (96%).Then this acid is dissolved among the DMF of 150mL, add (1R, 2S)-1-amino-2-vinyl cyclopropane-carboxylic acid carbethoxy hydrochloride (2.61g, 13.6mmol), N-methylmorpholine (2.5mL, 22.6mmol) and HATU (5.2g, 13.7mmol).At room temperature, at N ₂Stirred reaction mixture is 16 hours in the atmosphere, then vacuum concentration.Residue is distributed between the buffer (biphthalate) of ethyl acetate and pH value 4.Use saturated NaHCO ₃The solution washing organic facies, dry (MgSO ₄), filtering, vacuum concentration obtains bullion.Carry out flash chromatography (60%-80% ethyl acetate/hexane), obtain 6.0g (98%) 1-{ [1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (S) carbonyls]-(1R)-amino-2 (S)-vinyl-cyclopropane carboxylic acid acetoacetic ester white solid. ¹H NMR (500MHz, CD ₃0D) δ 1.25 (t, J=7.2Hz, 3H), 1.33-1.80 (m, 10H), 1.46 (s, 9H), 2.09 (m, 3H), 2.25 (m; 2H), 3.76 (m, 2H), 4.14 (m, 2H), 4.27 (dd, J=8.5,5.2Hz, 1H), 4.50 (m; 2H), 4.94 (d, J=10.1Hz, 1H), 5.01 (dd, J=17.1,1.8Hz, 1H), 5.11 (dd, J=10.4; 1.8Hz, 1H), 5.30 (d, J=15.6Hz, 1H), 5.80 (m, 2H), 8.57 (s, 1H); LC-MS (method A, retention time: 3.21min), MS m/z 522 (M ⁺+ 1).

Step 3:1-{ [1-(2-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4-(tert-butyl group-dimethyl-silanyloxy base)-pyrrolidine-2-carbonyl]-amino }-preparation of 2-vinyl cyclopropane carboxylic acid acetoacetic ester.

To 1-{ [1-(2 (S)-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4 (R)-hydroxyl-pyrrolidine-2 (S) carbonyls]-(1R)-amino }-2 (S)-vinyl-cyclopropane carboxylic acid acetoacetic ester (1.5g; 2.87mmol) the 10mLDMF mixture in add imidazoles (0.25g; 3.67mmol) and the tert-butyl group-dimetylsilyl chlorine (516mg, 3.44mmol).Mixture was at room temperature stirred two days.The vacuum concentration reactant mixture is dissolved in residue in the ethyl acetate then.With this solution of water washing, use dried over mgso, filter, vacuum concentration obtains the bullion solid.With purification by flash chromatography (with 20% ethyl acetate/hexane eluting), obtain 1.43g (78%) 1-{ [1-(2-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4-(tert-butyl group-dimethyl-silanyloxy base)-pyrrolidine-2-carbonyl]-amino }-2-vinyl cyclopropane carboxylic acid acetoacetic ester white solid. ¹H NMR (300MHz, CD ₃OD) δ 0.10 (s, 6H), 0.89 (s, 9H), 1.22 (m, 3H), 1.31-1.48 (m, 16H), 1.50-1.75 (m; 3H), 2.06 (m, 3H), 2.11-2.33 (m, 2H), 3.70 (m, 2H), 4.03-4.19 (m, 2H); 4.21 (m, 1H), 4.45 (t, J=7.87Hz, 1H), 4.59 (m, 1H), 4.91 (d; J=9.15Hz, 1H), 4.98 (d, J=17.20Hz, 1H), 5.08 (dd, J=10.25,1.83Hz; 1H), 5.27 (dd, J=17.38,1.65Hz, 1H), 5.65-5.87 (m, 2H) .LC-MS (method A, retention time: 4.00min); MS m/z 636 (M ⁺+ 1).

Step 4:14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] preparation of 19-7-alkene-4-carboxylic acid, ethyl ester

To 1-{ [1-(2-t-butoxycarbonyl amino-ninth of the ten Heavenly Stems-8-alkene acyl)-4-(tert-butyl group-dimethyl-silanyloxy base)-pyrrolidine-2-carbonyl]-amino }-2-vinyl-cyclopropane carboxylic acid acetoacetic ester (1.63g; 2.56mmol) the 640mL dichloromethane solution in add 215mg (0.26mmol) tricyclohexyl phosphine [1; 3-two (2; 4,6-three [benzal] ruthenous chloride (IV).With mixture reflux 15 minutes.The vacuum concentration residue is then used the flash column chromatography purification, with 30% ethyl acetate/hexane eluting.For sample is further decoloured, once more with the bullion chromatographic isolation,, obtain 1.5g (96%) 14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 with 50% ether/hexane eluting ^4,6] 19-7-alkene-4-carboxylic acid, ethyl ester white solid. ¹HNMR(500MHz，CD ₃Cl)δ0.06(s，3H)，0.07(s，3H)，0.86(s，9H)，1.18-1.24(m，6H)，1.34-1.64(m，14H)，1.86-1.96(m，3H)，2.02-2.09(m，1H)，2.11-2.17(m，1H)，2.19-2.28(m，1H)，2.57-2.63(m，1H)，3.50-3.54(m，1H)，3.71(dd，J＝10.22，6.26Hz，1H)，4.06-4.17(m，2H)，4.52-4.58(m，2H)，4.75(d，J＝8.55?Hz，1H)，5.21(t，J＝9.92Hz，1H)，5.35(d，J＝7.63Hz，1H)，5.45-5.50(m，1H)，6.94(s，1H)；

LC-MS (method A, retention time: 3.88min), MS m/z 608 (M ⁺+ 1).

Step 5:14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] preparation of 19-7-alkene-4-carboxylic acid

To 14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-4-carboxylic acid, ethyl ester (1.5g, and adding powder lithium hydroxide monohydrate in the solution of THF 2.47mmol) (4mL), methanol (1mL) and water (2mL) mixed solvent system (1.0g, 50mmol).In room temperature, at N ₂Stirred pale yellow syrup 4 hours in the atmosphere.Vacuum concentrated mixture then, and residue distributed between ether and water.Remove the ether phase, handle water with 1N HCl, till pH value 4.With this acid solution of ethyl acetate extraction (three times).With the dry (MgSO of the ethyl acetate extract that merges ₄), filtering, vacuum concentration obtains 1.2g (84%) 14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-4-carboxylic acids white solid. ¹H NMR (300MHz, CD ₃OD) 0.12 (s, 6H), 0.89 (s, 9H), 1.23-1.64 (m, 17H); 1.70-1.87 (m, 1H), 1.90-2.49 (m, 6H), 3.70-3.80 (m, 1H); 3.83-3.90 (m, 1H), 4.28-4.36 (m, 1H), 4.47-4.55 (m, 1H); 4.65 (s, 1H), 5.30-5.39 (m, 1H), 5.53-5.62 (m, 1H); LC-MS (method A, retention time: 3.69min), MS m/z580 (M ⁺+ 1).

Step 6: [18-(tert-butyl group-dimethyl-silanyloxy base)-4-cyclopropane sulfonyl amino carbonyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl]-preparation of carbamic acid tertiary butyl ester.

With 14-t-butoxycarbonyl amino-18-(tert-butyl group-dimethyl-silanyloxy base)-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] (500mg 0.86mmol) is dissolved among the 25mL THF 19-7-alkene-4-carboxylic acid, and (180mg 1.12mmol) handles with CDI.(take every caution against error, use the glass drying oven of oven dry, avoid moistening, N keeps dry ₂Atmosphere).After reaction mixture refluxed 2 hours, it is cooled to room temperature, with the cyclopropyl sulfonamide (135mg, 1.12mmol) and DBU (170mg, 1.12mmol) sequential processing.Reactant mixture was at room temperature stirred 4 hours, remove THF through rotary evaporation.Residue is distributed between the buffer of ethyl acetate and pH value 4.Dry (MgSO ₄) organic facies, filtering, vacuum concentration obtains bullion.Use purification by flash chromatography (with 33% ethyl acetate/hexane eluting) then, obtain 300mg (51%) [18-(tert-butyl group-dimethyl-silanyloxy base)-4-cyclopropane sulfonyl amino carbonyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl]-carbamic acid tertiary butyl ester white solid. ¹HNMR (300MHz, CD ₃OD) δ 0.07 (s, 3H), 0.08 (s, 3H), 0.85 (s, 9H), 0.87-1.49 (m, 21H), 1.73-1.95 (m; 3H), and 2.08-2.16 (m, 1H), 2.25-2.36 (m, 2H), 2.42-2.56 (m, 1H), 2.85-2.93 (m, 1H); 3.65-3.74 (dd, J=10.61,3.66Hz, 1H), 3.89 (d, J=10.25Hz, 1H), 4.34 (m, J=9.70; 9.70Hz, 1H), 4.43 (t, J=7.87Hz, 1H), 4.57 (s, 1H), 4.94-5.01 (m, 1H); 5.10 (d, J=8.78Hz, 1H), 5.66-5.75 (m, 1H), 6.55 (s, 1H), 10.13 (s, 1H); LC-MS (method A, retention time: 3.81min), MSm/z 683 (M ⁺+ 1).

Step 7: (4-cyclopropane sulfonyl amino carbonyl-18-hydroxyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl)-preparation of carbamic acid tertiary butyl ester.

To [18-(tert-butyl group-dimethylsilyl oxygen base)-4-cyclopropane sulfonyl amino carbonyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl]-the carbamic acid tertiary butyl ester (330mg, add in 25mL THF mixture 0.48mmol) tetrabutylammonium fluoride (150mg, 0.54mmol).Reactant mixture was at room temperature stirred 18 hours, then remove THF through rotary evaporation.Residue is distributed between ethyl acetate and water.Dry (MgSO ₄) organic facies, filtering, vacuum concentration obtains bullion.Through grinding its purification, obtain 200mg (73%) (4-cyclopropane sulfonyl amino carbonyl-18-hydroxyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 then with hexane ^4,6] 19-7-alkene-14-yl)-carbamic acid tertiary butyl ester white solid. ¹HNMR (500MHz, CD ₃Cl) δ 1.87-1.64 (m, 21H), 1.70-1.98 (m, 3H), 2.15-2.56 (m, 5H), 2.85-2.94 (m, 1H); 3.71 (d, J=13.91Hz, 1H), 4.10-4.26 (m, 2H), 4.51 (t, J=7.87Hz; 1H), 4.62 (s, 1H), 4.98 (m, 1H), 5.06 (d, J=8.78Hz; 1H), 5.64-5.71 (m, 1H), 6.72 (s, 1H), 10.24 (s, 1H); LC-MS (method A, retention time: 2.85min), MS m/z 569 (M ⁺+ 1).

Step 8:

At 0 ℃, to (4-cyclopropane sulfonyl amino carbonyl-18-hydroxyl-2,15-dioxo-3,16-diaza-three ring [14.3.0.0 ^4,6] 19-7-alkene-14-yl)-the carbamic acid tertiary butyl ester (1.00g, and the solid Dess-Martin reagent of a share of adding in dichloromethane 1.76mmol) (50mL) solution (1.64g, 3.87mmol).Remove after the ice bath, at room temperature stirred the mixture 3 hours, with 2-propanol (2mL) cancellation, extra stirring 1 hour, vacuum concentration.Use the ethyl acetate grinding residues, filter concentrated filtrate.Use the dichloromethane grinding residues, filter.Filtrating is concentrated into dried.Through quick column purification residue, at first use 2: 1,1: 1 hexane-acetone eluting then, obtain needed product white solid (670mg, 67%).LC-MS (retention time: 2.42min, method B), MS m/z 567 (M ⁺+ H).

Step 9:

In 70 ℃ of oil baths, under high vacuum condition, will contain magnesium bromide etherate (183mg, 2 neck flasks heating 0.71mmol) 4 hours.After being cooled to room temperature, and the adding tert-butyl group (6S, 13aS, 14aR, 16aS, Z)-14a-(cyclopropyl sulfuryl amino formoxyl)-2,5; 16-trioxy--1,2,3,5,6,7,8; 9,10,11,13a, 14,14a; 15,16, and 16a-16 hydrogen cyclopropyl [e] pyrrolo-[1,2-a] [1,4] diaza cyclopentadecylene-6-aminocarbamic acid esters (40mg, 0.071mmol).Use ice-cooled flask.Add anhydrous THF (2mL) through syringe.With the faint yellow serosity that forms stirred overnight energetically at room temperature.In refrigerative this flask, dropwise add with dry ice-propanone phenyl lithium (1.8M, in THF, 0.39mL, 0.71mmol).Under this temperature, stirred the mixture 2 hours, and then bath was replaced with ice bath, stirred in addition 1 hour.Use saturated NH ₄Cl cancellation mixture is used ethyl acetate extraction.With the isolating organic layer of brine wash, use MgSO ₄Drying is filtered, and concentrates.With hexane (5mL) grinding residues, filter.Concentrated filtrate through preparation HPLC purification, obtains the needed product white solid of 9.5mg (21%). ¹H NMR (CD ₃OD) δ 1.01-1.21 (m, 4H), 1.28-1.39 (m, 2H), 1.45 (s, 9H), 1.49-1.52 (m, 4H); 1.63-1.72 (m, 2H), 1.80-1.91 (m, 2H), 1.98-2.04 (m, 1H), 2.30-2.35 (m, 1H); 2.44-2.55 (m, 2H), 2.70-2.74 (m, 1H), 2.94-2.97 (m, 1H), 4.06-4.08 (m, 1H); 4.31-4.39 (m, 2H), 4.60-4.63 (m, 1H), 5.21-5.29 (m, 1H), 5.65-5.71 (m; 1H), and 7.31-7.34 (m, 1H), 7.38-7.42 (m, 2H), 7.63-7.65 (m, 2H); LC-MS (retention time: 2.74min, method B), MS m/z 627 (M ⁺+ H-H ₂O), 571 (M ⁺+ H-H ₂The O-t-butyl).

Embodiment 89: the preparation of chemical compound 3

Use and prepare chemical compound 3 with chemical compound 2 identical methods.In 70 ℃ of oil baths, under high vacuum condition, will contain magnesium bromide etherate (183mg, 2 neck flasks heating 0.71mmol) 4 hours.After being cooled to room temperature, and the adding tert-butyl group (6S, 13aS, 14aR, 16aS, Z)-14a-(cyclopropyl sulfuryl amino formoxyl)-2,5; 16-trioxy--1,2,3,5,6,7,8; 9,10,11,13a, 14,14a; 15,16, and 16a-16 hydrogen cyclopropyl [e] pyrrolo-[1,2-a] [1,4] diaza cyclopentadecylene-6-aminocarbamic acid esters (40mg, 0.071mmol).Use ice-cooled flask.Add anhydrous THF (2mL) through syringe.With the faint yellow serosity that forms stirred overnight energetically at room temperature.

At-78 ℃, to contain the 4-bromo biphenyl (165mg, dropwise add in another 2 neck flask (flask B) of THF 0.71mmol) (4mL) solution n-BuLi (2.5M, in hexane, 0.28mL, 0.71mmol), under this temperature with the solution stirring that forms 1 hour.

At-78 ℃, change among the flask A that is cooled to-78 ℃ through the inclusions of sleeve pipe with flask B.The little yellow solution stirring that under this temperature, will form 2 hours is then done-acetone bath with the ice bath replacement, stirs in addition 1 hour.Use saturated NH ₄Cl cancellation mixture is used ethyl acetate extraction.With the isolating organic layer of brine wash, use MgSO ₄Drying is filtered, and concentrates.With hexane (5mL) grinding residues, filter.Concentrated filtrate through preparation HPLC purification, obtains the needed product white solid of 4.1mg (8%). ¹H NMR (CD ₃OD) δ 0.87-1.10 (m, 4H), 1.24-1.38 (m, 2H), 1.45 (s, 9H), 1.49-1.52 (m, 4H), 1.70-1.84 (m; 4H), and 1.99-2.10 (m, 1H), 2.22-2.35 (m, 1H), 2.40-2.60 (m, 2H), 2.70-2.80 (m, 1H); 2.90-2.99 (m, 1H), 4.08-4.11 (m, 1H), 4.31-4.41 (m, 2H), 4.61-4.69 (m, 1H), 5.23-5.32 (m; 1H), and 5.61-5.70 (m, 1H), 7.35-7.38 (m, 1H), 7.43-7.48 (m, 2H), 7.62-7.70 (m, 6H); LC-MS (retention time: 2.74min, method B), MS m/z 703 (M ⁺+ H-H ₂O), 647 (M ⁺+ H-H ₂The O-t-butyl).

Biological study

In the disclosure, use HCV NS3/4A protease compound enzyme to test and test, and prepare as follows, operate and verify based on the HCV replicon of cell:

The generation of recombinant HCV NS3/4A protease complex

Be described below, form HCV NS3 protease complex derived from BMS strain, H77 strain or J4L6S strain.Form these purified recombinant body proteins, be used for homology test (seeing below), thereby provide active compound of the present disclosure how to suppress the indication of HCV NS3 proteolytic activity.

From Dr.T.Wright, San Francisco Hospital obtains the serum of HCV infected patient.Found full-length cDNA (complement DNA) template by dna fragmentation (reverse transcription-pcr (RT-PCR) by serum RNA (ribonucleic acid) obtains), use based on the selected primer of homology between other genotype 1a strain through the HCV genome (BMS strain) of design.By the mensuration of whole genome sequence, according to people's such as Simmonds classification, with genotype 1a distribute to the HCV self reliance type (referring to P Simmonds, KA Rose; S Graham, SW Chan, F McOmish; BCDow, EA Follett, PL Yap and H Marsden; J.Clin.Microbiol., 31 (6), 1493-1503 (1993)).The aminoacid sequence of non-structural region, NS2-5B, shown with the homogeny of HCV genotype 1a (H77)＞97% and with the homogeny of genotype 1b (J4L6S) 87%.Obtain infective clone body H77 (1a genotype) and J4L6S (1b genotype) by R.Purcell (NIH), and sequence is disclosed in, and (AAB67036 is referring to Yanagi among the Genbank; M., Purcell, R.H.; Emerson; S.U. and Bukh, J.Proc.Natl.Acad.Sci.U.S.A.94 (16), 8738-8743 (1997); AF054247, referring to Yanagi, M., St Claire, M., Shapiro, M., Emerson, S.U., Purcell, R.H. and Bukh, J, Virology 244 (1), 161-172 (1998)).

H77 and J4L6S strain are used for the preparation of recombinant NS3/4A protease complex.The recombinant HCV NS3/4A protease complex (amino acid/11 027 to 1711) of these strains is carried out dna encoding; According to the described method of people such as P.Gallinari carry out (referring to Gallinari P, Paolini C, Brennan D; Nardi C; Steinkuhler C, De Francesco R.Biochemistry.38 (17): 5620-32, (1999)).In brief, in 3 of NS4A coding region ' end adds three lysine solubilising afterbodys.Convert the locational cysteine of P1 of NS4A-NS4B cracking site (amino acid/11 711) into glycine, to avoid the proteolysis cracking of lysine label.In addition, at amino acid position 1454, utilize PCR to cause the sudden change of cysteine to serine, to prevent the self-dissolving cracking in NS3 helicase zone.Clone modification D NA fragment in pET21b bacterial expression vector (Novagen), and according to people such as P.Gallinari institute description scheme (referring to Gallinari P, Brennan D; Nardi C, Brunetti M, Tomei L; Steinkuhler C; De Francesco R., J Virol.72 (8): 6758-69 (1998)) remodeling (Invitrogen) is expressed the NS3/4A complex with e. coli strains BL21 (DE3).In brief, at 20 ℃, with the isopropyl ss-D-1-thiogalactoside (IPTG) of 0.5 mM (mM) with NS3/4A protease complex induced expression 22 hours.Typical fermentation (1 liter (L)) obtains about 10 gram (g) moistening cell ointment.Cell is resuspended in (the dissolving buffer is made up of following: 25mM N-(2-ethoxy) piperazine-N '-(2 ethane sulfonic aicd) (HEPES) in dissolving buffer (10mL/g); PH value 7.5; 20% glycerin, 500mM sodium chloride (NaCl), 0.5%Triton X-100; 1 mcg/ml (" μ g/mL ") lysozyme, 5mM magnesium chloride (MgCl ₂), 1 μ g/ml DnaseI, 5mM beta-mercaptoethanol (β ME), protease inhibitor-do not contain ethylenediaminetetraacetic acid (EDTA) (Roche)), 4 ℃ of homogenize and cultivated 20 minutes.At 4 ℃, with the homogenate supersound process, and through under 235000g, clarifying in ultra centrifugal 1 hour.Imidazoles is joined in the supernatant, and the final concentration to 15mM is adjusted to 8.0 with pH value.The crude protein extract is loaded on nitrilotriacetic acid nickel (Ni-NTA) post, and this post is with buffer B (25mM HEPES, pH 8.0,20% glycerin, 500mM NaCl, 0.5%Triton X-100,15mM imidazoles, 5mM β ME) pre-equilibration.Flow velocity filling sample with 1mL/min.Buffer C (identical with buffer B, as just to contain 0.2%TritonX-100) with 15 column volumes washs this post.Buffer D (C is identical with buffer, just contains the 200mM imidazoles) eluted protein with 5 column volumes.

Collection contains the part of NS3/4A protease complex, and is loaded on the desalting column Superdex-S200, and this post is with buffer D (25mM HEPES, pH 7.5,20% glycerin, 300mM NaCl, 0.2%Triton X-100,10mM β ME) pre-equilibration.Flow velocity filling sample with 1mL/min.Collection contains the fraction of NS3/4A protease complex, and is concentrated into about 0.5mg/ml.Conclude that through SDS-PAGE and mass spectral analysis the purity of NS3/4A protease complex (derived from BMS, H77 and J4L6S strain) is greater than 90%.Enzyme-80 ℃ of storages, is thawed on ice, dilute before in the test buffer, using.

The FRET peptide test of monitoring HCV NS3/4A proteolytic activity

The purpose of this in vitro tests be measure disclosure chemical compound to HCV NS3 protease complex (derived from BMS strain, H77 strain or J4L6S strain, inhibition as stated).This test provides active compound of the present disclosure how to suppress the index of HCV NS3 proteolytic activity.

In order to monitor HCV NS3/4A proteinase activity, use NS3/4A peptide substrate.This substrate is that (resonance energy shifts depsipeptide substrate (Resonance Energy TransferDepsipeptide Substrate) to RET S1; AnaSpec, Inc.cat#22991) (FRET peptide) is described in Anal.Biochem.240 (2): 60-67 (1996) by people such as Taliani.For HCV NS3 protease, the sequence of this peptide just has ester bond rather than amido link at cracking site loosely based on the natural cracking site of NS4A/NS4B.This peptide also contains near the fluorescence donor EDANS of an end of peptide with near the receptor DABCYL of another end.Shift (RET) through the intermolecular resonance energy between donor and receptor, fluorescence that can the quencher peptide, but when NS3 protease cracking peptide, product is from the release of RET quencher stage, and the fluorescence of donor becomes high-visible.

Under the condition that does not have or exist disclosure chemical compound, cultivate peptide substrate with one in three recombinant NS3/4A protease complex.Through using the formation of Cytofluor Series 4000 monitoring fluorescence reaction in real time product, measure the inhibition effect of chemical compound.

Reagent is following: obtain HEPES and glycerin (ultrapure) from GIBCO-BRL.Obtain dimethyl sulfoxine (DMSO) from Sigma.Obtain beta-mercaptoethanol from Bio Rad.

Test buffer: 50mM HEPES, pH value 7.5; 0.15M NaCl; 0.1%Triton; 15% glycerin; 10mM β ME.Substrate: 2 μ M final concentrations (being obtained from the 2mM stock solution, in DMSO)-20 ℃ of storages.HCV NS3/4A protease type 1a (1b), 2-3nM final concentration (being obtained from 5 μ M stock solutions, in 25mM HEPES (pH value 7.5), 20% glycerin, 300mM NaCl, 0.2%Triton-X100,10mM β ME).For the chemical compound that has near the ultimate potentiality of test, through in the test buffer, adding 50 μ g/ml bovine serum albumin (Sigma), and final protease concentration is reduced to 300pM, can make test sensitiveer.

In 96 hole polystyrene black plates (being obtained from Falcon), make an experiment.25 μ l NS3/4A protease complex (in the test buffer), 50 μ l chemical compound of the present disclosure (in 10%DMSO/ test buffer) and 25 μ l substrate (in the test buffer) are contained in each hole.On the identical test plate, prepare tester (not containing chemical compound).Multienzyme complex was mixed 1 minute with chemical compound or tester solution, then cause enzymatic reaction through adding substrate.Use CytofluorSeries 4000 (Perspective Biosystems) will test the plate reading immediately.Instrument is set, thereby reads the emission of 340nm and exciting of 490nm at 25 ℃.Reaction was carried out about 15 minutes usually.

Suppress with the following equation calculated percentage:

100-[(δF _inh/δF _con)×100]

Wherein δ F is the change in fluorescence in the range of linearity of curve.Nonlinear curve fitting is used for inhibition-concentration data, and through utilizing Excel XLfit software, user's formula y=A+ ((B-A)/(1+ ((C/x) ^D))) calculates 50% valid density (IC ₅₀Value).

The specificity test

Carry out the specificity test, with the external selectivity of explanation disclosure chemical compound when suppressing HCV NS3/4A protease (comparing) with other serine or cysteine proteinase class.

With respect to various serine stretch protein enzymes, measure the specificity of disclosure chemical compound: human neutrophil Elastase (HNE), Pancreas Sus domestica gland Elastase (PPE) and human pancreas Chymotrypsin and a kind of thiol protease: human hepatic tissue protease B.In all cases; Use the previous 96 hole plate formula schemes of describing (use and have amino-methyl-coumarin (AMC) particular substrate of fluorescence) (PCT patent application No.WO 00/09543), carry out some improvement for the serine stretch protein enzyme test for every kind of enzyme.All enzymes are from Sigma, and EMDbiosciences buys, and substrate is obtained from Bachem, Sigma and EMDbiosciences.

According to the potential energy of chemical compound, compound concentration changes between 100 to 0.4 μ M.Cause each enzyme test through substrate being joined in the enzyme inhibitor (at room temperature cultivated in advance 10 minutes, and on cytofluor, measure, be hydrolyzed into 15% conversion ratio).

The final condition of each test is following:

50mM three (hydroxymethyl) aminomethane hydrochloride (Tris-HCl) pH value 8,0.5M sodium sulfate (Na ₂SO ₄), 50mM NaCl, 0.1mM EDTA, 3%DMSO, 0.01%Tween-20 (containing 5 μ M LLVY-AMC) and 1nM Chymotrypsin.

50mM Tris-HCl (pH value 8.0), 50mM NaCl, 0.1mM EDTA, 3%DMSO, 0.02%Tween-20,5 μ M succ-AAPV-AMC and 20nM HNE or 8nM PPE;

100mM NaOAC (sodium acetate) pH value 5.5,3%DMSO, 1mM TCEP (three (2-carboxyethyl) phosphonium salt hydrochlorate), 5nM cathepsin B (before using, activatory proenzyme material in the buffer that contains 20mM TCEP) and 2 μ M Z-FR-AMC (in water, diluting).

The percentage ratio that uses computes to suppress:

[1-((UV _inh-UV _blank)/(UV _ctl-UV _blank))]×100

Nonlinear curve fitting is used for inhibition-concentration data, through utilizing Excel XLfit computed in software 50% valid density (IC ₅₀Value).

The generation of HCV replicon

Utilize the method described in following to set up the complete cell system of HCV replicon: Lohmann V, Korner F, Koch J, Herian U, Theilmann L, Bartenschlager R., Science 285 (5424): 110-3 (1999).This system can make us estimate the effect of HCV protease chemical compound to the HCV rna replicon.In brief, and use Lohmann article (accession number: the HCV strain 1b sequence AJ238799), through Operon Technologies; Inc. (Alameda; CA) synthetic HCV cDNA uses standard molecular biological technique, then at plasmid pGem9zf (+) (Promega; Madison, WI) middle assembling total length replicon.Replicon is made up of following: (i) HCV 5 ' UTR; 12 aminoacid of itself and capsid protein condense; (ii) neomycin phosphotransferase gene (neo) (iii) is obtained from the IRES of encephalomyocarditis virus (EMCV) and (iv) HCV NS3 to NS5B gene and HCV 3 ' UTR.With DNA s linearisation,, use T7 MegaScript transcript reagent box (Ambion, Austin, TX) vitro synthesized RNA transcription product with ScaI according to the explanation of manufacturer.With the in vitro transcription product transfection of cDNA in human liver cancer cell line HUH-7.In the presence of selected marker thing neomycin (G418), obtain selection to cell that can structural expression of HCV replicon.Resultant as time passes cell line is endowed the characteristic of positive and negative chain RNA product and protein product.

HCV replicon FRET test

Carry out HCV replicon FRET test, with the inhibition effect of monitoring chemical compound that the disclosure is described to the HCV virus replication.Make the HUH-7 cell of structural expression of HCV replicon be grown in the improved Eagle culture medium of Dulbecco (DMEM) (Gibco-BRL) in, this culture medium contain 10% hyclone (FCS) (Sigma) with 1mg/ml G418 (Gibco-BRL).In 96 hole tissue culture sterile plates, before the darkness cell is sowed (1.5 * 10 ⁴Individual cells/well).In dilution plate (in test, the 0.5%DMSO final concentration), containing 4%FCS, 1: 100 penicillin/streptomycin (Gibco-BRL), 1: prepare chemical compound among the DMEM of 100L-glutamine and 5%DMSO and do not have the tester of chemical compound.Chemical compound/DMSO mixture is joined in the cell, cultivated 4 days at 37 ℃.After 4 days, at first use ALMA blue (Trek Diagnotstic Systems) to estimate the cytotoxicity of cell, with CC ₅₀Registration.Through in the medium of cultured cell, adding 1/10 ^ThThe ALMA of volume is blue, measures the toxicity (CC of chemical compound ₅₀).After 4 hours, read the fluorescence signal in each hole,, use CytofluorSeries 4000 (Perspective Biosystems) with the excitation wavelength of 530nm and the emission wavelength of 580nm.Use PBS (PBS) (3 times, 150 μ l) cleaning down plate then.With the solubility test agent dissolves of cell with 25 μ l; This reagent contains HCV protease substrate (the plain enzyme cell culture solubilising reagent (Promega#E153A) of 5X cell fluorescence); With distilled water diluting to 1X; NaCl joins the 150mM ultimate density, and FRET peptide substrate (top enzyme test is described) is diluted to 10 μ M ultimate densities (using the 2mM raw material, in 100%DMSO).Then plate is put into Cytofluor 4000 instruments, this instrument is excited/the 490nm emission to 340nM surely, 21 circulation automatic modes are carried out the plate reading with kinetics model.According to IC ₅₀The described method of pH-value determination pH is carried out EC ₅₀PH-value determination pH.

HCV replicon luciferase indication test

As the test of next, in replicon luciferase indication test, proved the EC that obtains by replicon FRET test ₅₀The pH-value determination pH data.The use of replicon luciferase indication test at first is described (Krieger N, Lohmann V, and BartenschlagerR, J.Virol.75 (10): 4614-4624 (2001)) by people such as Krieger.3 of the humanization form of utilization embedding cDNA coding Renilla luciferase gene and direct and luciferase gene ' hold condensed catenation sequence improved FRET and tested described replicon constructs.Use is arranged in the Ascl restriction site (the direct upper reaches of neomycin marker gene) of nuclear, and this embedding is introduced in the replicon constructs.Also can be in the position 1179 (serine is to isoleucine) cause adaptive mutation (Blight KJ, Kolykhalov, AA, Rice, CM, Science 290 (5498): 1972-1974).Form the stable cell lines of this HCV replicon constructs of structural expression, as stated.Test described method according to having following improved HCV replicon FRET, set up luciferase indication test.At 37 ℃/5%CO ₂After 4 days, use Promega Dual-Glo luciferase pilot system, the Renilla uciferase activity of analysis of cells in the incubator.From each hole of containing cell, remove medium (100 μ l).In remaining 50 μ l media, add 50 μ l Dual-Glo luciferase reagent, at room temperature plate was shaken 10 minutes to 2 hours.Then Dual-Glo Stop & Glo reagent (50 μ l) is joined in each hole, at room temperature shook plate once more 10 minutes to 2 hours.Use the fluorescence program, on Packard TopCount NXT with the plate reading.

The percentage ratio that uses computes to suppress:

This value is processed figure, use XLfit to analyze, obtain EC ₅₀Value.

Note,, can find the structure of chemical compound in this article through patent working example numbering and the patent compound number that use is shown in Table 2.

Table 2

The embodiment numbering	Compound number	IC50(nM)	EC50(nM)
				87	1	18	639
88	2	9	389
				89	3	2	14

It is that significantly the disclosure is not limited to the illustrative embodiment of aforesaid explanation to those skilled in the art, and under the condition that does not deviate from necessary characteristic of the present disclosure, it can be contained with other particular form.Therefore expectation; Embodiment is considered to explain illustrative in all fields; Rather than restrictive, should therefore be intended to comprise the implication of the equivalent that falls into claim and all changes in the scope with reference to additional claim rather than previous embodiment.

Claims

1. the chemical compound of formula (I):

Or its officinal salt, wherein

R ¹Be selected from alkoxyl, hydroxyl and-NHSO ₂R ⁷

R ^2aAnd R ^2bBe independently selected from hydrogen and methyl;

R ³Be selected from thiazolinyl, alkyl, aryl, aralkyl, cycloalkyl, (cycloalkyl) alkyl, heterocyclic radical and heterocyclic radical alkyl;

R ⁴It is hydroxyl;

r ⁵Be selected from hydrogen, alkyl and cycloalkyl;

R ⁷Be selected from alkyl, aryl, cycloalkyl, (cycloalkyl) alkyl, heterocyclic radical and-NR ^aR ^bWherein the cycloalkyl moiety of cycloalkyl and (cycloalkyl) alkyl is optional is independently selected from following group replacement by one, two or three: thiazolinyl, alkoxyl, alkoxyalkyl; Alkyl, aralkyl, aryl carbonyl; Cyanic acid, cycloalkenyl group, (cycloalkyl) alkyl; Halogen, halogenated alkoxy, haloalkyl and (NR ^eR ^f) carbonyl; And R wherein ^aAnd R ^bBe independently selected from hydrogen, alkoxyl, alkyl, aryl, aralkyl, cycloalkyl, (cycloalkyl) alkyl, haloalkyl, heterocyclic radical and heterocyclic radical alkyl; And R wherein ^eAnd R ^fBe independently selected from hydrogen, alkyl, aryl, aralkyl and heterocyclic radical; Wherein the aryl moiety of aryl, aralkyl and heterocyclic radical are optional is replaced by one or two substituent group that is independently selected from alkoxyl, alkyl and halogen; With

Q is C _3-9Saturated or unsaturated chain optionally contains one to three and is independently selected from O, S (O) _mAnd NR ⁹Hetero atom, wherein m is 0,1 or 2, and R ⁹Be selected from hydrogen, alkoxyl, alkoxy carbonyl group, alkyl, alkyl-carbonyl, alkyl sulphonyl, amino carbonyl, aryl sulfonyl, cycloalkyl, (cycloalkyl) alkyl, cycloalkyl oxy, di alkylamino group carbonyl, di alkylamino group carbonylic alkyl, haloalkyl and heterocyclic radical carbonyl.

2. the chemical compound of claim 1, it is the chemical compound of formula (II):

Or its officinal salt, wherein

R ¹Be-NHSO ₂R ⁷

R ^2aAnd R ^2bBe hydrogen;

R ⁴It is hydroxyl;

R ⁵Be hydrogen;

R ⁶It is alkoxy carbonyl group;

3. the compound or pharmaceutically acceptable salt thereof of claim 1, wherein R ¹Be-NHSO ₂R ⁷

4. claim 2 or 3 compound or pharmaceutically acceptable salt thereof, wherein R ⁷It is cycloalkyl.

5. claim 1,2 or 3 compound or pharmaceutically acceptable salt thereof, wherein Q contains the heteroatomic C of zero ₆Unsaturated chain.

6. claim 1 or 2 chemical compound are selected from following

Or its officinal salt.

7. pharmaceutical composition, it comprises each compound or pharmaceutically acceptable salt thereof and pharmaceutically suitable carrier of claim 1-6.

8. the compositions of claim 7 further comprises at least a other chemical compound with anti-HCV activity.

9. the compositions of claim 8, wherein at least a other chemical compound is interferon or ribavirin.

10. the compositions of claim 8, wherein at least a other chemical compound be selected from interleukin 2, interleukin 6, interleukin 12, can increase the chemical compound that the response of 1 type helper T lymphocyte forms, interfering RNA, antisense RNA, miaow Kui Mote, ribavirin, inosine 5 '-monophosphate dehydrogenase inhibitor, amantadine and rimantadine.

11. each compound or pharmaceutically acceptable salt thereof of claim 1-6 is used for treating the purposes of the medicine that patient HCV infects in preparation, this purposes comprise give the patient treatment effective dose claim 1-6 each compound or pharmaceutically acceptable salt thereof.

12. the purposes of claim 11 further comprises giving at least a other chemical compound with anti-HCV activity, this chemical compound is before giving each compound or pharmaceutically acceptable salt thereof of claim 1-6, give afterwards or with it simultaneously.

13. the purposes of claim 12, wherein at least a other chemical compound is interferon or ribavirin.