CN101084236A - excitatory amino acid prodrugs - Google Patents

Abstract

The present invention relates to synthetic excitatory amino acid prodrugs and methods of making the same. The invention further relates to methods of use thereof, and pharmaceutical compositions comprising the compounds for the treatment of neurological and psychiatric disorders.

Description

Prodrugs of excitatory amino acids

The invention provides synthetic prodrugs of excitatory amino acids (formula I compound) and preparation method thereof.The invention further relates to its using method, and comprise pharmaceutical composition formula I compound, that be used for the treatment of nervous system disorders and mental disorder.

Background of invention

Neural system or mental disorder, for example the treatment of anxiety disorder is relevant with the selectively activate of the excitatory amino acid receptor of metabolic pattern.For example, the U.S. Patent No. 5,688,826 (' 826 patent) that on November 18th, 1997 published discloses (+)-4-amino-2-sulphonyl dicyclo [3.1.0] hexane-4, and the 6-dicarboxylic acid is as active mGluR2 receptor stimulant.In addition, the U.S. Patent No. 5,958,960 of publishing on September 28th, 1999 (" 960 patents) (+)-2-amino-4-fluorine dicyclo [3.1.0] hexane-2 is disclosed, the 6-dicarboxylic acid is as active mGluR2 receptor stimulant.

The invention provides the prodrug forms of mGluR2 receptor agonist compounds, it can improve the interior drug effect of body of each parent drug, and produces higher parent drug oral cavity contact.Compounds represented of the present invention keep the security and the curative effect of previously disclosed mGluR2 receptor stimulant, and strengthen the best progress of oral administration biaavailability.

Series number is that the PCT application of PCT/US01/45866 and PCT/US02/00488 discloses synthetic prodrugs of excitatory amino acids and preparation method thereof.

Summary of the invention

The invention provides compound or its pharmaceutically useful salt of formula I.

Wherein,

A is H-(Q) _p-;

When occurring at every turn, Q independently is selected from aminoacyl;

P is the integer of 1-10;

X is O, S, SO, SO ₂Or CR ³R ⁴

R ³Be fluorine, X ' OR ⁵, SO ₃H, tetrazolium-5-base, CN, PO ₃R ⁶ ₂, hydroxyl or NO ₂, and R ⁴Be hydrogen; Or R ³And R ⁴Represent fluorine separately; Or R ³And R ⁴Together representative=O ,=NOR ⁷Or CR ⁸R ⁹Or R ³Or R ⁴In one represent amino and another representation carboxy; Or R ³Represent N ₃, (CH ₂) mCOOR ^5a, (CH ₂) mPO ₃R ^6a ₂, NHCONHR ^5bOr NHSO ₂R ^5c, and R ⁴Represent hydrogen; Or R ³And R ⁴Representative=CHCOOR together ^5b,=CHPO ₃R ^6a ₂Or=CHCN;

X ' represents key, CH ₂Or CO;

M is the integer of 1-3;

R ⁵, R ^5a, R ^5b, R ^5c, R ⁷, R ⁸And R ⁹Independent is hydrogen atom; Optional (1-6C) alkyl that replaces; Optional (2-6C) thiazolinyl that replaces; Optional (2-6C) alkynyl that replaces; The optional aryl that replaces; The optional heteroaryl that replaces; Non-aromatic carbon ring group; Non-heteroaromatic group; With one or two monocyclic aryl or heteroaryl-condensed non-aromatic monocycle carbon ring group; Or with one or two monocyclic aryl or heteroaryl-condensed non-aromatic monocyclic heterocycles group;

R ⁶And R ^6aIndependent hydrogen or (1-6C) alkyl represented;

R ¹⁰Be hydrogen or fluorine; With

R ¹¹Be hydrogen, fluorine or hydroxyl;

The present invention also provides formula I compound, wherein substituting group such as top definition, but compound is not that wherein X is CR ³R ⁴, R wherein ³Be fluorine, and R ⁴Be hydrogen, p is 1, and Q is the compound of L-alanyl; Or its pharmaceutically useful salt.

The present invention also provides formula I compound, and wherein A is H-(Q) _p-; When occurring at every turn, Q independently is selected from aminoacyl; P is the integer of 1-3; X is O, S, SO, SO ₂Or CR ³R ⁴R ³Be fluorine or hydroxyl, and R ⁴Be hydrogen; Or R ³And R ⁴Representative=O together; R ¹⁰Be hydrogen or fluorine; And R ¹¹Be hydrogen, fluorine or hydroxyl; Or its pharmaceutically useful salt.

The present invention also provides formula I compound, and wherein A is H-(Q) _p-; Q is the L-alanyl; P is 1; X is CR ³R ⁴R ³Be fluorine and R ⁴Be hydrogen; R ¹⁰Be hydrogen; And R ¹¹Be hydrogen; Or its pharmaceutically useful salt.

What deserves to be mentioned is that formula I compound contains at least four unsymmetrical carbons.The present invention includes all stereoisomeric forms in any ratio of formula I compound, comprise enantiomer that each is independent and composition thereof, for example the prodrug forms of disclosed compound in ' 826 patents; as 1SR, 4RS, 5RS; 6RS-4-amino-(2-alkylsulfonyl dicyclo [3.1.0] hexane)-4, the 6-dicarboxylic acid.

Another aspect of the invention provides pharmaceutical preparation, and it comprises in conjunction with the formula I compound of pharmaceutically useful carrier, thinner or excipient or its pharmaceutically useful salt.

Another aspect of the invention provides the method for the metabotropic glutamate receptor that cAMP connects in a kind of patient body of influence, and it comprises the patient who the formula I compound administration of medicine effective quantity is subjected to synthetic excitatory amino acid neurotransmission in needs.The present invention also provides a kind of purposes of formula I compound, is used to make the medicine that influences the metabotropic glutamate receptor of cAMP coupling connection in the patient body.

Another aspect of the invention provides the method for the formula II compound that gives significant quantity, comprises the patient who the formula I compound administration of medicine effective quantity is subjected to synthetic excitatory amino acid neurotransmission in needs.The present invention also provides a kind of purposes of formula I compound, is used to make the medicine of the formula II compound that gives significant quantity.

Another aspect of the invention provides a kind of method of the patient's of treatment nervous system disorders, comprises that formula I compound administration with medicine effective quantity is in the patient of this treatment of needs.The present invention also provides a kind of purposes of formula I compound, is used to make the medicine of treatment patient nervous system disorders.

Another aspect of the invention provides a kind of method of the patient's of treatment mental disorder, comprises that formula I compound administration with medicine effective quantity is in the patient of this treatment of needs.The present invention also provides a kind of purposes of formula I compound, is used to make the medicine of treatment patient mental disorder.

Formula I compound can prepare according to the method for the heterogeneous ring compound that is similar to the known preparation similar of chemical field, or according to novel method preparation described herein.The useful intermediates that this method and being used for prepares above-mentioned formula I compound step below describes, and wherein, except as otherwise noted, otherwise the implication of general group is definition herein.

The invention provides a kind of method of preparation I compound, comprise aminoacyl acidylate formula compound (ii) with corresponding formula III:

Pg ^N-A- (III)

Pg wherein ^NBe nitrogen-protecting group group, A such as above-mentioned definition;

Then, for above-mentioned any one step,, then remove blocking group if use blocking group that functional group is protected;

Then, for above-mentioned any one step: the pharmaceutically useful salt of formula I compound if desired, this formula I compound that then makes the alkali form with provide the acid of pharmaceutically useful counterion to react; Or, make this compound of sour form and pharmaceutically useful cationic alkali reaction is provided for the formula I compound that contains acid moieties; Or for the zwitterionic compound of formula I, the formula I compound of neutralizing acid adduct form or alkali adduct form; Or adopt any other traditional method.

The present invention also provides formula I compound, and wherein X is CH ², R ¹⁰Be fluorine, other variable such as above-mentioned definition.

Detailed Description Of The Invention

Found that The compounds of this invention is the useful prodrug of metabotropic glutamate receptor selective agonist compound, therefore and be used for the treatment of central nervous system disease, as nervous system disorders, nerve degeneration disease for example, and as antipsychotic drug, anxiolytic, medicine withdrawal, thymoleptic, anticonvulsive drug, anodyne and antiemetic.

What deserves to be mentioned is that formula I compound contains at least four unsymmetrical carbons, three are positioned on the cyclopropane ring, and one is the alpha-carbon atom of amino acid group.Therefore, compound of the present invention can be present in optical purity form, racemic form or the non-enantiomer mixture, or therefrom separates.

Amino acid moiety preferably has the natural amino acid configuration, that is, and and with respect to the L-configuration of D-Glycerose.

The present invention includes the pharmaceutically useful salt of formula I compound.These salt can combine existence with the acidity or the basic moiety of molecule, and can be used as acid-adducting salt, and primary, secondary, uncle or quaternary ammonium salt, basic metal or alkaline earth salt exist.Usually, acid-adducting salt can prepare by the reaction of acid with formula I compound.In addition, acid-adducting salt also can prepare like this: make the compound (protected intermediates) of penultimate stride and suitable normal acid-respons generate corresponding salt form, this salt and then prepared in reaction formula I compound or other salt.Basic metal and alkaline earth salt are usually by the metal of hope

The crystal formation of some specific salts brings the specific formulation advantage.Non-crystal amorphous compound can have water absorbability.Sometimes more wish to obtain the medical compounds of crystal formation, because it demonstrates favourable solid state properties.

The acid that is generally used for forming this salt comprises mineral acid, for example hydrochloric acid, Hydrogen bromide, nitric acid, sulfuric acid or phosphoric acid, or organic acid, as organic carboxylic acid, for example oxyacetic acid, toxilic acid, hydroxymaleic acid, FUMARIC ACID TECH GRADE, oxysuccinic acid, tartrate, citric acid, Whitfield's ointment, acetoxybenzoic acid, or organic sulfonic acid, 2-ethylenehydrinsulfonic acid, tosic acid, methylsulfonic acid, naphthalene-2-sulfonic acid, Phenylsulfonic acid or ethyl sulfonic acid.

Except pharmaceutically useful salt, the present invention also comprises other salt.It can be used as the intermediate of purifying compounds, or as the intermediate for preparing other pharmaceutically useful acid-adducting salt, or be used for discriminating, sign or purifying.

Among the present invention, formula I compound comprises its solvate.Especially, formula I compound comprises its hydrate.

And the present invention has also predicted the prodrug of disclosed fluorine cpd among International Application PCT/JP99/03984, PCT/JP99/00324 and the PCT/JP01/05550.Respectively referring to international publication WO/0012464, WO/9938839 and WO/0200605.For example, the present invention has predicted 1S, 2R, 5S, 6S-2-amino-6-fluoro-4-oxo dicyclo [3.1.0] hexane-2,6-dicarboxylic acid; 1S, 2R, 4S, 5S, 6S-2-amino-6-fluoro-4-hydroxyl dicyclo [3.1.0] hexane-2,6-dicarboxylic acid; 1S, 2R, 3R, 5S, 6S-2-amino-3-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid; And 1S, 2R, 3S, 5S, 6S-2-amino-6-fluoro-3-hydroxyl dicyclo [3.1.0] hexane-2,6-dicarboxylic acid.

Various physiological functions have demonstrated and have been subjected to stimulus effects excessive or excitatory amino acid transmission improperly.Believe that formula I compound of the present invention has the ability of the treatment Mammals various nervous system disorderss relevant with this symptom, comprise acute nervous system disorders, as the heart by-pass operation with the brain defective after transplanting, apoplexy, cerebral ischemia, spinal cord injuries receptor, head trauma, term anoxic, cardiac arrest and hypoglycemia nerve injury.Believe that formula I compound has the ability of the various chronic neurological disorders of treatment, dementia, eyes wound and retinopathy, cognitive illnesses and the congenital and drug-induced Parkinson's disease of bringing out as presenile dementia, Huntington Chorea, amyotrophic lateral sclerosis, acquired immune deficiency syndrome (AIDS).The present invention also provides the method for the treatment of these diseases, comprises that the patient to this treatment of needs gives formula I compound or its pharmaceutically useful salt of significant quantity.

The various nervous system disorderss relevant of other of formula I compounds for treating patient of the present invention with the L-glutamic acid dysfunction, comprise muscle spasm, convulsions, migraine, the urinary incontinence, pain, thirst for (for example, Nicotine, opiate, Cocaine, benzene phenodiazine  and alcohol), anxiety and relative disease, vomiting, cerebral edema, chronic pain and tardive dyskinesia through preceding agitation (PDD), psychosis (as schizophrenia), drug tolerance, withdrawal, drug withdrawal and medicine.Formula I compound also can be used as thymoleptic and anodyne.Therefore, the present invention also provides the method for the treatment of these diseases, comprises formula I compound or its pharmaceutically useful salt to patient's effective dosage of this treatment of needs.

Following definition is used to illustrate the meaning and the scope of various terms used herein.Essential Terms used herein have its common meaning.

Term " influence " refers to the agonist of formula II compound as excitatory amino acid receptor.Term " excitatory amino acid receptor " refers to metabotropic glutamate receptor, a kind of acceptor through GTP-conjugated protein and the coupling of cytological effect device.Term " metabotropic glutamate receptor of cAMP coupling connection " refers to suppress through coupling the metabotropic receptor of adenylate cyclase activity.

Term " nervous system disorders " refers to acute and chronic neurodegenerative disorders, comprise heart bypass operation and transplant after brain defective, cerebral ischemia (for example, cardiac arrest cause apoplexy), spinal cord injuries receptor, head trauma, presenile dementia, Huntington Chorea, amyotrophic lateral sclerosis, the acquired immune deficiency syndrome (AIDS) dementia of bringing out, term anoxic, hypoglycemia nerve injury, eye wound and retinopathy, cognitive illnesses and congenital and drug-induced Parkinson's disease.This term also comprises the neurological conditions that other is caused by the L-glutamic acid dysfunction, comprise muscle spasm, migraine, the urinary incontinence, drug tolerance, withdrawal, drug withdrawal and medicine thirst for (that is, opiate. benzene phenodiazine , Nicotine, Cocaine or alcohol), smoking cessation, vomiting, cerebral edema, chronic pain, sleep disease, convulsions, Tourette syndromes, ADHD (Attention Deficit Hyperactivity Disorder) and tardive dyskinesia.

Term " mental disorder " refers to acute and chronic mental illness disease, comprise schizophrenia, anxiety and relative disease (as, panic and pressure-dependent cardiovascular disorder), depressed, bipolarity is unusual, abalienation, compulsive disorder, generalized anxiety disorder, acute pressure disease and panic disease.

The term of Shi Yonging " significant quantity " refers to the amount or the dosage of compound herein, and based on the dosage of patient's single or multiple, its diagnosis or treatment to patient provides the effect that needs.

As those skilled in the art, the attending doctor is easy to determine its significant quantity by the result who uses currently known methods and observe under the analogue.In the significant quantity and dosage determined to drug compound, the attending doctor will consider several factors, includes but not limited to: mammal species; Its stature size, age and total healthy state; The disease specific that relates to; The reaction that relates to degree or severity individual patient of disease; The specific compound of administration; Administering mode; The bioavailability characteristics of drug-delivery preparation; The medicament taking method of selecting; The use of the pharmacotherapy of following; And other correlation circumstance.For example, can contain the about 300mg activeconstituents of the 5mg-that has an appointment usually every day in the dosage.Can carry out the administration of compound by all means, comprise oral, rectum, approach in skin, subcutaneous, intravenously, intramuscular, mouth or nose.In addition, compound can carry out administration by continuous transfusion.

The term of Shi Yonging " patient " refers to Mammals herein, as mouse, cavy, rat, dog or people.Should be appreciated that preferred patient is the people.

The term of Shi Yonging " is treated " (or " treatment ") and is comprised its usually implication of approval herein, it comprise prevention, prevent and treat, check and alleviate, stop or reversing the carrying out of ill disease.Like this, method of the present invention comprises therapeutic administration and preventive administration.

The generalization technics of Shi Yonging has its common implication herein.For example, term " (1-6C) alkyl " means the straight or branched group.(1-6C) example value of alkyl group comprises (1-4C) alkyl, as methyl, ethyl, propyl group, sec.-propyl, butyl and isobutyl-.Term " (2-6C) thiazolinyl " comprises (2-4C) thiazolinyl, as allyl group.Term " (2-6C) alkynyl " comprises (2-4C) alkynyl, as proyl.

Used term " optional replacement " in term " optional (1-6C) alkyl group that replaces ", " choosing (2-6C) alkenyl group that replaces wantonly " and " optional (2-6C) alkynyl group that replaces ", can there be one or more substituting groups in expression herein, preferred 1-3, described substituting group is selected from such atom and group: when it is present in the formula I compound, do not hinder formula I compound to regulate the function of metabotropic glutamate receptor.

Being present in the atom in optional (1-6C) alkyl group that replaces, optional (2-6C) alkenyl group that replaces and optional (2-6C) alkynyl group that replaces and the example of group is, the optional aromatic yl group that replaces, the optional heteroaryl groups that replaces, non-aromatic carbon ring group, non-heteroaromatic group, with one or two monocyclic aryl or heteroaryl-condensed non-aromatic monocycle carbon ring group and with one or two monocyclic aryl or heteroaryl-condensed non-aromatic monocyclic heterocycles group.

Term " heteroaryl " comprises that containing 1-4 is selected from heteroatomic aromatic series 5-6 unit's ring of oxygen, sulphur and nitrogen and contains 1-4 and be selected from heteroatomic 5-6 unit's ring and phenyl ring of oxygen, sulphur and nitrogen or contain 1-4 heteroatomic 5-6 unit that is selected from oxygen, sulphur and nitrogen and encircle the aromatic series bicyclic radicals that condenses composition.The example of heteroaryl is furyl, thienyl,  azoles base, different  azoles base, thiazolyl, isothiazolyl, imidazolyl, pyrimidyl, benzofuryl, benzothienyl, benzimidazolyl-, benzoxazol base, benzothiazolyl and indyl.

Term " aromatic group " comprises phenyl and Ppolynuclear aromatic carbocyclic ring, as naphthyl.

Used term " optional replacement " in term " optional replace heteroaryl group " and " the optional aromatic group that replaces ", can there be one or more substituting groups in expression herein, described substituting group is selected from such atom and group: when it is present in the formula I compound, do not hinder formula I compound to regulate the function of metabotropic glutamate receptor.

Being present in optional heteroaryl group that replaces or the atom in the optional aromatic group that replaces and the example of group is, amino, hydroxyl, nitro, halo, (1-6C) alkyl, (1-6C) alkoxyl group, (1-6C) alkylthio, carboxyl, (1-6C) carbalkoxy, formamyl, (1-6C) alkanoylamino, (1-6C) alkyl sulphonyl, (1-6C) alkyl sulfonyl amino, the optional phenyl that replaces, phenoxy group, thiophenyl, benzenesulfonyl, benzenesulfonyl amino, tolylsulfonyl amino, (1-6C) fluoroalkyl and (1-6C) Fluoroalkyloxy.Object lesson is amino, hydroxyl, fluorine, chloro, bromo, iodo, methyl, methoxyl group, methylthio group, carboxyl, kharophen, methylsulfonyl, nitro, ethanoyl, phenoxy group, thiophenyl, benzenesulfonyl, methanesulfonamido and trifluoromethyl.

The optional aromatic group that replaces be exemplified as the 1-naphthyl, the 2-naphthyl, phenyl, the 2-xenyl, the 3-xenyl, the 4-xenyl, the 2-hydroxy phenyl, the 3-hydroxy phenyl, the 4-hydroxy phenyl, the 2-fluorophenyl, the 3-fluorophenyl, the 4-fluorophenyl, 2, the 4-difluorophenyl, 3, the 4-difluorophenyl, pentafluorophenyl group, the 2-chloro-phenyl-, the 3-chloro-phenyl-, the 4-chloro-phenyl-, 2, the 4-dichlorophenyl, 3, the 4-dichlorophenyl, 3-chloro-4-fluorophenyl, 3, the 5-dichlorophenyl, the 2-bromophenyl, the 3-bromophenyl, the 4-bromophenyl, the 2-aminomethyl phenyl, the 3-aminomethyl phenyl, the 4-aminomethyl phenyl, the 2-p-methoxy-phenyl, the 3-p-methoxy-phenyl, the 4-p-methoxy-phenyl, 2, the 3-Dimethoxyphenyl, 2, the 5-Dimethoxyphenyl, 3, the 4-Dimethoxyphenyl, 3, the 5-Dimethoxyphenyl, the 2-trifluoromethyl, the 3-trifluoromethyl, the 4-trifluoromethyl, 2-fluoro-3-trifluoromethyl, 3-trifluoromethyl-4-fluorophenyl, 3-trifluoromethyl-5-fluorophenyl, 2-fluoro-5-trifluoromethyl, the 2-Phenoxyphenyl, the 3-Phenoxyphenyl, 3-carboxyl phenyl and 4-carboxyl phenyl.

Term " non-aromatic carbon ring group " comprises monocyclic groups, (3-10C) cycloalkyl for example, as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl, the ring octyl group, ring nonyl or ring decyl, with the fused polycycle group, for example 1-adamantyl or 2-adamantyl, 1-decahydro naphthyl (decalyl), 2-decahydro naphthyl, 4a-decahydro naphthyl, dicyclo [3,3,0] suffering-1-base,-2-base or-the 3-base, dicyclo [4,3,0] ninth of the ten Heavenly Stems-the 1-base,-2-base,-3-base or-the 7-base, dicyclo [5,3,0] last of the ten Heavenly stems-the 1-base,-2-base,-3-base,-4-base,-8-base or-9-base and dicyclo [3,3,1] ninth of the ten Heavenly Stems-the 1-base,-2-base,-3-base or-the 9-base.

Term " non-heteroaromatic group " comprises and contains one or two heteroatomic aromatic series 4-7 unit ring, this heteroatoms is selected from oxygen, sulphur and nitrogen, for example azetidine-1-base or-the 2-base, tetramethyleneimine-1-base,-2-base or-the 3-base, piperidines-1-base,-2-base,-3-base or-the 4-base, six hydrogen azepine-1-base,-2-base,-3-base or-the 4-base, trimethylene oxide-2-base or-the 3-base, tetrahydrofuran (THF)-2-base or-the 3-base, tetrahydropyrans-2-base,-3-base or-the 4-base, the hexahydrooxepin-2-base,-3-base or-the 4-base, propylene sulfide-2-base or-the 3-base, tetramethylene sulfide-2-base or-the 3-base, tetrahydric thiapyran-2-base,-3-base or-the 4-base, six hydrogen thia  (hexahydrothiepin)-2-bases,-3-base or-the 4-base, piperazine-1-base or-the 2-base, morpholine-1-base,-2-base or-the 3-base, thiomorpholine-1-base,-2-base or-the 3-base, tetrahydropyrimidine-1-base,-2-base,-4-base or-the 5-base, tetrahydroglyoxaline-1-base,-2-base or-the 4-base, imidazolidine-1-base,-2-base or-the 4-base,  azoles quinoline-2-base,-3-base,-4-base or-the 5-base,  azoles alkane-2-base,-3-base,-4-base or-the 5-base, thiazoline-2-base,-3-base,-4-base or-the 5-base, or thiazolidine-2-Ji,-3-base,-4-base or-the 5-base.

Term " with one or two monocyclic aryl or heteroaryl-condensed non-aromatic monocycle carbon ring group " comprises with phenyl ring or contains the individual oxygen that is selected from of 1-4, the first ring of heteroatomic aromatic series 5-6 condensed (3-10C) cycloalkyl of sulphur and nitrogen, for example indanyl, 1,2,34-tetraline-1-base or-the 2-base, 5,6,7,8-tetrahydroquinoline-5-base,-6-base,-7-base or-the 8-base, 5,6,7,8-tetrahydroisoquinoline-5-base,-6-base,-7-base or-the 8-base, 4,5,6,7-tetrahydro benzo thiophene-4-base,-5-base,-6-base or-the 7-base, dibenzo [2,3,6,7] ring heptan-1-base or-the 4-base, dibenzo [2,3,6,7] ring heptan-4-alkene-1-base or-the 4-base, or 9-fluorenyl.

Term " with one or two monocyclic aryl or heteroaryl-condensed non-aromatic monocyclic heterocycles group " comprises that containing one or two is selected from oxygen, sulphur and nitrogen assorted former in 4-7 unit ring, itself and phenyl ring or contain 1-4 and be selected from oxygen, the aromatic series 5-6 unit ring of sulphur and nitrogen heteroatom condenses, for example 2,3-dihydrobenzopyrans-2-base,-3-base or-the 4-base, xanthene-9-base, 1,2,3,4-tetrahydroquinoline-1-base,-2-base,-3-base or-the 4-base, 9,10-acridan-9-base or-the 10-base, 2,3-thiochroman-2-base,-3-base or-the 4-base, or dibenzothiopyrans-4-base.

Use, by " Pg herein ^N" expression term " nitrogen-protecting group group " refer to be intended to protect or block nitrogen groups carries out undesirable reaction in synthesis step those groups.The selection that suitable used nitrogen-protecting group is rolled into a ball will be depended on the condition of protected reaction subsequently, and it is known for those of ordinary skills.T.W.Greene and P.G.M.Wuts, ProtectiveGroups In Organic Synthesis, the 3rd edition (John Wiley﹠amp; Sons, New York (1999)) in nitrogen-protecting group group commonly used is disclosed.Preferred nitrogen-protecting group is rolled into a ball and is tertbutyloxycarbonyl.

Use, by " Pg herein ^C" term " carboxy protective group " of expression is when referring to that other functional group of compound reacts, and is usually used in blocking or protecting a kind of carboxylates derivatives of hydroxy-acid group.Object lesson comprises, for example methyl, ethyl, the tertiary butyl, benzyl, methoxymethyl, trimethyl silyl etc.Other example of this group can be at T.W.Greene and P.G.M.Wuts, Protecting Groupsin Organic Synthesis, the 3rd edition (JohnWiley﹠amp; Sons, New York (1999)) in find.Preferred carboxy protective group is methyl and ethyl.Use traditional method that does not influence the molecule other parts that this ester is decomposed.

The group that term " hydroxy-protective group " expression organic chemistry filed those of skill in the art understand, Greene and P.G.M.Wut. describe in the 2nd chapter.Representational hydroxy-protective group comprises, for example, and the ether group of ether group, replacement, isopropyl ether group, benzene and substituted benzene ether group, benzyl and substituted benzyl ether group, alkyl silyl ether group, ester protecting group etc.The type requirements of used hydroxy-protective group is not strict; as long as the deutero-oh group is stable in the reaction of subsequently other position of intermediate molecule, and selected removing and the residue molecule that comprises any other hydroxy-protective group is not produced interference at the appropriate time.

Term " aminoacyl " refers to that by amino acid derived and next aminoacyl this amino acid is selected from the group of the natural or alpha-non-natural amino acid composition of definition herein.Natural amino acid can be neutrality, positivity or negativity according to the substituting group on the side chain." neutral amino acids " refers to contain the amino acid of uncharged side chain substituents.Exemplary neutral amino acids comprises L-Ala, Xie Ansuan, leucine, Isoleucine, proline(Pro), phenylalanine, tryptophane, methionine(Met), glycine, Serine, Threonine, halfcystine and glutamine and asparagine." positive amino acid " refers to that side chain substituents is electropositive amino acid under the physiological pH.Exemplary positive amino acid comprises Methionin, arginine and Histidine." negative amino acid " refers to that side chain substituents is clean electronegative amino acid under the physiological pH.Exemplary negative amino acid comprises aspartic acid and L-glutamic acid.Preferred amino acids is an a-amino acid.Most preferred amino acid is to be the stereochemical a-amino acid of L-on the alpha-carbon atom.Exemplary natural a-amino acid is Xie Ansuan and Isoleucine, proline(Pro), phenylalanine, tryptophane, methionine(Met), glycine, Serine, Threonine, halfcystine, asparagine, glutamine, Methionin, arginine, Histidine, aspartic acid and L-glutamic acid.

" alpha-non-natural amino acid " refers to the amino acid of free nucleic acid codon.The example of alpha-non-natural amino acid comprises, for example, and the D-isomer of above-mentioned natural a-amino acid; Aib (aminobutyric acid), β Aib (3-aminoisobutyric acid), Nva (norvaline), β-Ala, Aad (2-aminoadipic acid), β Aad (3-aminoadipic acid), Abu (2-aminobutyric acid), Gaba (γ-An Jidingsuan), Acp (6-aminocaprolc acid), Dbu (2, the 4-DAB), the alpha-amino group pimelic acid, TMSA (trimethyl silyl-Ala), alle (not-Isoleucine), Nle (nor-leucine), uncle-Leu, Cit (citrulline), Orn, Dpm (2,2 '-diaminopimelic acid), Dpr (2, the 3-diaminopropionic acid), α-or β Nal, Cha (cyclohexyl-Ala), oxyproline, Sar (sarkosine), the O-methyltyrosine, phenylglycocoll etc.; Cyclic amino acids; N ^a-alkylation amino acid, wherein N ^a-alkylation amino acid is N ^a-(1-10C) alkyl amino acid, for example MeGly (N ^a-methylglycine), EtGly (N ^a-ethyl glycine) and EtAsn (N ^α-ethyl asparagine) and contain the amino acid of two side chain substituents on the alpha-carbon.Exemplary non-natural a-amino acid comprises D-L-Ala, D-leucine and phenylglycocoll.The UNC of biological chemical name joint committee (JCBN) suggestion of IUPAC-IUB is followed in the natural and alpha-non-natural amino acid that herein uses and the name of residue thereof, as it at european journal of biological chemistry (European Journal of Biochemistry), 138,9-37 (1984), set forth in " Nomenclature and Symbolism for Amino Acid andPeptides (Recommendations, 1983) ".Different as the name of the amino acid that uses in this specification sheets and the appended claims and residue thereof and abbreviation and the name of above-mentioned custom, then will clarify different names and abbreviation.

Though all formula I compounds all are useful active mGluR2 receptor stimulant, preferably some compound.Following paragraph has defined preferred kind.

A) Q is glycyl, alanyl, valyl, leucyl, isoleucyl, prolyl, phenylalanyl, tyrosyl, tryptophyl, methionyl, lysyl or seryl.

B) Q is an alanyl.

C) Q is a methionyl.

D) P is 1.

E) P is 2.

F) X is SO ₂

G) X is CR ³R ⁴

H) R ³Be fluorine, and R ⁴Be hydrogen.

I) R ³Be hydroxyl, and R ⁴Be hydrogen.

J) R ³And R ⁴Representative=O together.

K) R ¹⁰Be hydrogen.

L) R ¹⁰Be fluorine.

M) R ¹¹Be hydrogen.

N) compound is a free alkali.

O) compound is a salt.

P) compound is a hydrochloride.

Q) compound is a mesylate.

R) compound is an esilate.

S) compound is a tosylate.

Earlier paragraphs can combine the in addition preferred classes of compounds of definition.

Formula I compound is useful for the treatment mammalian diseases, and preferred Mammals is behaved.

Compound of the present invention can use the several different methods preparation, and the some of them method has been described in the following scheme.The particular order of each step that preparation I compound is required depends on the relative instability of synthetic specific compound, initial compounds and replacement part.For the sake of clarity, some substituting groups in the following scheme may be removed, and this substituting group is not intended to limit by any way the instruction to scheme.Understand substituent R in those of ordinary skill as ability ¹⁵And R ¹⁶Represent suitable side chain, to form the aminoacyl of wishing.

If can not be commercially available, then can prepare necessary starting raw material in the following scheme by being selected from the organic and method heterocyclic chemistry standard technique, this technology and known, structural similitude compound synthetic similar, this method is description to some extent in preparation and embodiment, and also comprises novel method.

Scheme 1

As shown in top scheme 1, formula I compound transforms through enzyme or hydrolytic process in vivo.Especially, the crystalline form of formula I compound can be according to the approach preparation of general introduction in the following scheme 2.

Scheme 2

The formula peptide acyl compound (iii) of diester protection in THF or THF/ water solvent, with suitable alkali, obtains the formula peptide acyl compound (iv) that diacid is protected as lithium hydroxide or sodium hydroxide effect at appropriate solvent.Formula (iv) compound can be carried out deprotection with suitable acid in appropriate solvent.But the hydrochlorate of the corresponding diacid peptide acyl compound shown in this condition production I salt, it can be amorphous solid, or directly is crystalline solid, wherein X " represent corresponding negatively charged ion.For amorphous solid, it subsequently can be by crystallizing out in the appropriate solvent.Thereby introduce cationic substance by for example sodium acetate reagent and can form carboxylate salt.At last, can obtain amphoteric substance with suitable alkaline purification crystal salt compound.

For example, the formula peptide acyl compound (iv) of diacid protection is handled the amorphous solid hydrochloride that can obtain deprotection with hydrogen chloride gas in appropriate solvent.The amorphous hydrochloride compound can crystallize out from acetone and water and generate the crystalline hydrochloride compound then.For the crystalline solid of direct formation, reaction mixture can obtain crystal salt after filtration.Can obtain amphoteric substance with the sodium-hydroxide treatment crystalline hydrochloride.Those of ordinary skills it should be understood that formula I compound can prepare and intermediate shown in not separating through a step.

Scheme 3

Use suitable coupling reagent with formula III compound acylation formula diester (ii), to obtain the peptide acyl compound of formula diester protection (iii).In addition, use the acyl chlorides of formula III compound also can obtain this conversion.

Suitable peptide coupling reagent comprises dicyclohexyl carbodiimide (DCC), 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDC), isobutyl chlorocarbonate, chlorine phosphono diphenyl ester, 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), two (2-oxo-3- oxazolidinyl) phosphonyl chloride and benzotriazole-1-base oxidation three (dimethylamino)  phosphofluoric acid ester.

Scheme 4

In the top scheme 4, with suitable carboxy protective reagent, the diacid compounds as catalysis hydrochloric acid or thionyl chloride and methyl alcohol or Ethanol Treatment formula II has obtained corresponding formula diester (ii).In addition, nitrogen-protection reagent can be used, as BOC earlier ₂O handles formula II compound, with formula (i) compound that obtains nitrogen-protection.Next can be at alkali, exist down as salt of wormwood, use carboxy protective reagent, as, methyl iodide is handled formula (i) compound, and then uses the nitrogen deprotecting regent, and example hydrochloric acid or trifluoroacetic acid processing formula (i) compound are with (ii) compound of acquisition formula.

In addition, those of ordinary skills it should be understood that depends on X, also may need suitable protection reagent.For example, if X represents CR ³R ⁴, R ³Representation hydroxy, and R ⁴Represent hydrogen, those of ordinary skills it should be understood that and carry out needing suitable hydroxy-protective group before above-mentioned any one scheme so.

Formula II compound is known in the art.For example, can be at United States Patent (USP) 5,688, find the preparation of these compounds in 826 (' 826 patents) and 5,958,960 (' 960 patents).

At the route of synthesis of formula II compound, disclosed method before various improvement have been carried out.This improvement comprises sulphur and pure oxidation, and the optical resolution of the various intermediates that describe below.

First kind of improvement relates to that " 826 patents the 8th hurdle, the conversion described in capable and the 7th hurdle of 22-34 since the 33rd hurdle (formula V), relates to the formula VII compound of ' 826 patents

Oxidation to form the formula V compound of ' 826 patents.

Find that in many method for oxidation well known in the prior art, preferably sulphur trioxide-pyridine complex or trifluoroacetic anhydride and DMSO's combines.

Secondly, for the formula III compound that splits ' 826 patents,

Wherein, R ²The representation carboxy group, with reference to the 8th hurdle, preferred (R)-α-Jia Jibianji amine and quinine have been found in capable and the 6th hurdle the 1st row beginning of 3-7.Preferred especially (R)-α-Jia Jibianji amine.

Once more; find that with reference to ' 826 patents the 8th hurdle, 39-53 is capable; X is that the formula III sulfide of sulphur carries out oxidation and forms when X is the formula III compound of alkylsulfonyl in ' 826 patents in ' 826 patents, and preferred alkaline water solution system and hydrogen peroxide combine use with catalyzer.

Following embodiment further illustrates compound of the present invention and synthetic method thereof.This embodiment is not intended in office where face limits the scope of the invention, and also should not understand like this.All tests all are that the direct draught at drying nitrogen or argon gas carries out.Except as otherwise noted, all solvents and reagent are all commercially available and use with purchased form.Exsiccant tetrahydrofuran (THF) (THF) can be obtained by distillation in sodium or the benzophenone sodium ketyl before use.Obtain on the Bruker of the 500MHz AvanceII bay-500, on the Bruker Avance I bay-200 of 200MHz or on the Varian Inova/Varian 300/Varian400 of 500MHz proton magnetic resonance (PMR) ( ¹H NMR) spectrogram.On Agilent MSD/B instrument, use acetonitrile/ammonium acetate aqueous solution to carry out electrospray ionization mass spectrum analysis (ESI) as moving phase.On VG ZAB-2SE instrument, carry out free atom bombardment mass spectroscopy (FABMS).Use VG 70SE or Varian MAT 731 instrument to carry out FDMS analysis (FDMS).Measure specific rotation with Perkin-Elmer 241 polarimeters.Usually use the linear gradient solvent of pointing out in the article to separate in the enterprising circumstances in which people get things ready for a trip spectrum of Waters Prep 500 LC.Usually use thin-layer chromatography (TLC) monitoring reaction to carry out whether fully.Use E.Merck Kieselgel60 F ₂₅₄Plate, 5cm * 10cm, 0.25mm is thick, carries out thin-layer chromatographic analysis.Use UV and chemical measure (plate is immersed in the cerous molybdate ammonium solution [75g ammonium molybdate in 500mL 10% aqueous sulfuric acid and 4g cerous sulfate (IV)], on hot-plate, heats then) to detect spot.According to Still etc., Still, Kahn and Mitra be at J.Org.Chem.43, and the methods of describing in 2923 (1978) are carried out flash chromatography.Go up the ultimate analysis of determining carbon, hydrogen and nitrogen at control device company 440 elemental analysers (ControlEquipment Corporation 440 Elemental Analyzer), or by Universidad Complutense Analytical Center (Facultad deFarmacia, Madrid, Spain) measure.In the opening glass capillary of Gallenkamp warm air bath fusing point instrument or B ü chi fusing point instrument, measure fusing point, and the fusing point that records is uncorrected.

The abbreviation of using among the embodiment, symbol and term have following meanings.

The Ac=ethanoyl

The Anal.=ultimate analysis

The ATR=attenuated total internal reflection

Bn or Bzl=benzyl

The Bu=butyl

The BOC=tertbutyloxycarbonyl

Calcd=calculates

D ₂The O=water-d2

DCC=dicyclohexyl carbodiimide

DCM=1, the 2-methylene dichloride

The DIBAL-H=diisobutyl aluminium hydride

The DMAP=4-dimethyl aminopyridine

The DMF=dimethyl formamide

The DMSO=dimethyl sulfoxide (DMSO)

The DSC=dsc

DEC=N-ethyl-N ' N '-dimethylaminopropyl carbodiimide hydrochloride

The ES=electrospray ionization mass spectrum

The Et=ethyl

EtOH=ethanol

FAB=fast atom bombardment (mass spectroscopy)

The FDMS=FDMS

The FTIR=fourier transform infrared spectrometry

HOAt=1-hydroxyl-7-azepine benzotriazole

The HOBt=1-hydroxybenzotriazole

The HPLC=high performance liquid chromatography

The HRMS=high resolution mass spec

The i-PrOH=Virahol

The IR=infrared spectra

The L=liter

The Me=methyl

MeOH=methyl alcohol

The MPLC=medium pressure liquid chromatography

The MP=fusing point

The MTBE=t-butyl methyl ether

The NBS=N-bromo-succinimide

The NMR=nucleus magnetic resonance

PC-TLC=preparative centrifugation thin-layer chromatography

The Ph=phenyl

The p.o.=oral administration

The i-Pr=sec.-propyl

Rochelle ' s Salt=Seignette salt

The rt=room temperature

The SM=starting raw material

The TBS=t-butyldimethylsilyl

The TEA=triethylamine

The Temp.=temperature

The TFA=trifluoroacetic acid

The THF=tetrahydrofuran (THF)

The TLC=thin-layer chromatography

The t-BOC=tert-butoxycarbonyl

General method A

EDC coupling between amine and N-BOC-(L)-amino acid

With the amino dialkyl (1.0 equivalent) of initiator (scheme 3, formula ii compound) be suspended under the nitrogen atmosphere, in the exsiccant methylene dichloride.Order adds corresponding N-Boc-(L)-amino acid (1.5-2.0 equivalent), EDC (1.5-2.0 equivalent), HOBt (1.5-2.0 equivalent) and dimethyl aminopyridine (DMAP, 0.1-0.2 equivalent).If not explanation is arranged in addition, then stirred reaction mixture reacts completely until judging with TLC under the room temperature.Use the ethyl acetate diluted reaction mixture, and use saturated NaHCO successively ₃The aqueous solution and/or NaHSO ₄The aqueous solution and salt water washing.After dried over sodium sulfate and the vacuum-evaporation, use suitable elutriant (being generally ethyl acetate/hexane) to make with extra care crude product resistates (formula iii compound) with silica gel chromatography.

General method B

Acid anhydrides coupling between amine and N-BOC-(L)-amino acid isobutyric anhydride

At-20 ℃, under the nitrogen atmosphere, in the solution that corresponding N-Boc-(L)-amino acid (1.5 equivalent) forms, add N-methylmorpholine (NMM, 1.5 equivalents, 1mL CH in exsiccant methylene dichloride (10mL) ₂Cl ₂In), drip isobutyl chlorocarbonate (IBCF, 1.5 equivalents, 5mL CH with certain speed afterwards ₂Cl ₂In), so that internal reaction temperature is no more than-15 ℃.Under-20 ℃, the gained reaction mixture was stirred 30 minutes, afterwards with certain speed add-20 ℃, (1S, 2S, 4S, 5R, 6R)-and 2-(2 '-amino-propionamido-)-4-hydroxyl-dicyclo [3.1.0] hexane-2, the solution that 6-diethyl dicarboxylate hydrochloride (1.0 equivalent) forms in methylene dichloride (10mL) is so that internal reaction temperature is no more than-15 ℃.After drip finishing, remove cooling bath and at room temperature stirred reaction mixture judge until TLC and react completely.Use the ethyl acetate diluted reaction mixture, and use saturated NaHCO successively ₃The aqueous solution, NaHSO ₄The aqueous solution and salt water washing.After dried over mgso and the vacuum-evaporation, use suitable elutriant (being generally hexane/ethyl acetate) to make with extra care the crude product resistates with silica gel chromatography.

General method C

Order is removed N-Boc and ester protecting group

Under the room temperature, in 1: 1 mixture of THF/2.5N LiOH (10-20 equivalent), stir corresponding N-Boc diester peptide derivant (formula iii compound, scheme 2) (1.0 equivalent) and reach 4 hours.The dilute with water reactant, and wash with ethyl acetate.Discard organic layer.With 1N HCl (NaCl is added aqueous phase to improve extractibility on demand) water being adjusted to pH is 2, and extracts N-boc dicarboxylic acid product (formula iv compound) fully with ethyl acetate.Merge all organism, salt water washing, MgSO ₄Drying, and vacuum concentration is to dry, with the spumescence solid carboxylicesters product that obtains to wish.Be dissolved in the ethyl acetate and be cooled to 0 ℃.Saturated with anhydrous HCl gas purging reaction mixture up to HCl.0 ℃ is stirred the gained reaction mixture down and reaches 4 hours.Filter under the nitrogen atmosphere, the peptide derivant (formula I compound) of complete deprotection is separated with hydrochloride, or reaction mixture is concentrated into drying, subsequently again with ethyl acetate or Et ₂O grinds and is condensed into white powder.Randomly, in order to remove residual solvent and excessive HCl, in water recombinant products, freeze the hydrochloride product that can obtain to wish with postlyophilization.

Preparation example 1

(1R, 4S, 5S, 6S) 4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-diethyl dicarboxylate

Under the room temperature, with 20 minutes with thionyl chloride (15.5mL, 212.6mmol) drop to (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4 in the slurries that 6-dicarboxylic acid (10g, 42.5mmol, U.S. Patent number 5,688,826) forms, is used the rinsing of 40mL ethanol subsequently in 100mL 2B ethanol.Slurries are heated to reflux and stir and spend the night.With 500MHz ¹HNMR (CD ₃OD) analyze spissated aliquots containig, the result shows that starting raw material and intermediate monoesters are fallen by completely consumed.Gained solution is cooled to room temperature, is concentrated into gluey residuum then.To further adding EtOAc (50mL) in the simmer down to solid gel, dilute with another part 94mL EtOAc then.(70mL) slowly adds in the mixture with 15% aqueous sodium carbonate, and rotates by hand with dissolving gradually, and the gained final pH is 7.95.Before extracting each layer gained yellow soda ash precipitation is filtered.With EtOAc (2 * 100mL) reextraction water layers.With salt solution (organic extract, drying (MgSO that 1 * 100mL) washing merges ₄), filter and vacuum concentration, thereby obtain faint yellow oily thing, its curing obtains the title compound (11.71g, 95% yield) into pale solid.

Recrystallization

With title compound (200mg) mixture heating up to 56 among the EtOAc (800 μ L) ℃, dissolve this moment.56 ℃ are stirred after 15 minutes down, and (1mL) drops in the solution with heptane.Turn off heating.Solution is cooled to 52 ℃, precipitating action takes place this moment.Further slurries have been formed in the dissolved process in cooling with heptane (600 μ L).Before the filtration, at room temperature stirred the gained slurries 1 hour, with heptane (2 * 500 μ L) washing, and 45 ℃ of following dried overnight, thereby obtain 145mg (73% rate of recovery) title compound, be white solid.

mp 80-83℃。

[α] ²⁵D-57.7°(c 1.04，CH ₃OH)。

500MHz ¹HNMR(CD ₃Cl ₃)δ4.31(q，2H，J＝7.0Hz)，4.20(m，2H)，3.78(d，1H，J＝15.0Hz)，3.36(dd，1H，J＝4.0，7.0Hz)，2.93(dd，1H，J＝4.0，7.0Hz)，2.81(d，1H，J＝15.0Hz)，2.46(t，1H，J＝4.0)，1.34(t，3H，J＝7.0)，1.30(t，3H，J＝7.0).

¹³C NMR(125MHz，CD ₃Cl ₃)δ171.68，168.57，63.26，62.42，59.96，56.06，43.78，32.25，22.49，14.31，14.25。

FTIR(ATR)3364.15(s)，1725.95(s)，1304.91(s)，1259.24(s)，1200.84(s)，1104.91(s)，1022.99(s)，896.45(s)，851.21(s)cm ^-1。

C ₁₁H ₁₇NO ₆The analytical calculation value of S: C, 45.35; H, 5.88; N, 4.81.Measured value: C, 45.02; H, 5.75; N, 4.82.

Preparation example 2

(1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-propionamido)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-diethyl dicarboxylate

Under-30 ℃, with 10 minutes with isobutyl chlorocarbonate (30.4mL, 234mmol) drop to the N-Boc-L-L-Ala (43.52g, 230mmol) and N-methylmorpholine (25.5mL is 232mmol) in the solution that forms in the 457mL methylene dichloride.Stirred the gained dilute slurry 30 minutes down at-25--30 ℃, this moment with clock time adding in 25 fens (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the solution that 6-diethyl dicarboxylate (63.90g, 219mmol, preparation example 1) forms in the 213mL methylene dichloride is so that temperature of reaction is no more than-25 ℃.After adding, remove cooling bath and also stirred at ambient temperature 60 minutes, this moment, temperature of reaction reached 19 ℃, and color becomes greenish orange look.With 350mL 1NHCl processing reaction thing and separate each layer.Use saturated NaHCO ₃The aqueous solution (1 * 350mL) and salt solution (1 * 350mL) washing organic layer, drying (Na ₂SO ₄), filter, and vacuum concentration is white foam shape thing (105.2g, 104%).

¹H NMR(300MHz，CDCl ₃)δ：7.62(brs，1H)，4.90(brd，1H，J＝7.1Hz)，4.34-4.10(m，6H)，3.39(ddd，1H，J＝7.2，3.9，1.0Hz)，3.00(dd，1H，J＝7.1，3.9Hz)，2.90(brd，1H，J＝14.9Hz)，2.43(t，1H，J＝4.1Hz)，1.46(s，9H)，1.31(m，9H，)。

¹³C NMR(75 MHz，CD ₃Cl ₃)δ：173.0，168.6，167.6，80.9，76.5，63.3，62.3，59.9，55.7，42.8，31.5，28.2，22.7，16.6，14.0，13.9。MS(ES)m/z 461.0[M-H] ^-。

Preparation example 3

(1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-propionamido)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dicarboxylic acid

Under the room temperature, to (1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-propionamido)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-diethyl dicarboxylate (181.4g, the 392mmol theoretical value, preparation example 2) adding 490mL (980mmol) 2N sodium hydroxide in the solution that in 292mL THF, forms. this two-phase mixture of vigorous stirring is 1.25 hours under the room temperature, react homogeneous phase carry out this moment. with 490mL ethyl acetate diluted mixture thing and separate each layer. and with 490mL ethyl acetate dilution water layer, and with concentrated hydrochloric acid mixture pH is reduced to 1.5. and separate each layer and also use 245mL ethyl acetate reextraction water layer.The dry organic layer (Na of getting up that merges ₂SO ₄), filter and concentrate, be the title compound of white foam shape thing thereby obtain 167.9g (105%).This material just is used for embodiment 1 and 2 without characterizing.

Preparation example 4

(1R, 4S, 5S, 6S)-and 4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

With thionyl chloride (6.2mL, 85.0mmol) drop to quick stirring (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4 is in the suspension that 6-dicarboxylic acid (10.0,42.5mmol, United States Patent (USP) 5,688,826) forms in MeOH (170mL, 5 ℃).After adding reaction mixture slowly is warming up to room temperature, under reflux temperature, is incubated 48 hours afterwards.Volatile matter is removed in decompression, and makes residuum at saturated NaHCO ₃Distribute between solution (200mL) and the ethyl acetate (400mL).Separate each layer, and with ethyl acetate (2 * 400mL) aqueous layer extracted.Use K ₂CO ₃Organic layer and concentrating under reduced pressure that dry merging is got up, thus 8.10g (30.8mmol) title compound obtained with 72% yield.

[α] ²³D＝-84°(c＝0.5，MeOH)。

C ₉H ₁₃NO ₆The analytical calculation value of S: C, 41.06; H, 4.98; N, 5.32.Measured value: C,

40.94；H，4.93；N，5.30。

MS(ES)m/z 264.0[M+H] ⁺.

Preparation example 5

(1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-3 '-phenyl-propionamido)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, use commercially available acquisition N-BOC-(L)-phenylalanine and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 4) is prepared.Reaction mixture refluxed is spent the night.Use PC-TLC, 4mmSiO ₂Rotor, (10% ethyl acetate/hexane-100% ethyl acetate) is refining.Acquisition 0.85g (88%, 1.67mmol) white foam shape thing.

[α] ²³D＝-35.2°(c＝0.45，CHCl ₃)。

¹H NMR(300MHz，CDCl ₃)δ1.43(9H，s)，2.38-2.40(1H，m)，2.86(1H，d，J＝15.0Hz)，2.91(1H，dd，J＝4.4，7.3Hz)，3.04(2H，d，J＝7.3Hz)，3.35-3.39(1H，m)，3.77(3H，s)，3.84(3H，s)，4.11(1H，d，J＝14.3)，4.30(1H，app.q，J＝7.3)，4.96(1H，bd，J＝6.6Hz)，6.96(1H，bs)，7.22-7.36(5H，m)。

C ₂₃H ₃₀N ₂O ₉S 0.1H ₂The analytical calculation value of O: C, 53.92; H, 5.94; N, 5.47.

Measured value: C, 53.62; H, 5.90; N, 5.28.

MS(ES)m/z 509.16[M-H]-；411.2 [M-Boc] ⁺。

Preparation example 6

(1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-3 ' S-methyl-valeryl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, use commercially available acquisition N-BOC-(L)-Isoleucine and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 4) is prepared.Reaction mixture refluxed is spent the night.Use PC-TLC, 4mmSiO ₂Rotor, (10% ethyl acetate/hexane-100% ethyl acetate) is refining.Acquisition 0.75g (83%, 1.57mmol) white foam shape thing.

[α] ²³D＝-32.65°(c＝0.49，CHCl ₃)。

¹H NMR(300 MHz，CDCl ₃)δ 0.91(3H，t，J＝7.3Hz)，0.93(3H，d，J＝6.6Hz)，1.10-1.18(1H，m)，1.46(9H，s)，1.42-1.52(1H，m)，1.81-1.86(1H，bm)，2.51(1H，t，J＝4.0Hz)，2.95(1H，d，J＝15.0Hz)，3.06(1H，dd，J＝4.4，7.3Hz)，3.43(3H，dd，J＝3.7，7.0Hz)，3.78(3H，s)，3.85(3H，s)，3.82-3.90(1H，m)，4.20(1H，d，J＝14.7Hz)，4.94(1H，d，J＝8.4Hz)，7.15(1H，bs)。

C ₂₀H ₃₂N ₂O ₉The analytical calculation value of S: C, 50.41; H, 6.77; N, 5.88.Measured value: C, 50.32; H, 6.92; N, 5.76.

MS(ES)m/z 475.1[M-H] ^-。

Preparation example 7

(1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-3 '-methyl-butyrylamino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, use commercially available acquisition N-BOC-(L)-Xie Ansuan and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 4) is prepared.Reaction mixture refluxed is spent the night.Use PC-TLC, 4mmSiO ₂Rotor, (10% ethyl acetate/hexane-100% ethyl acetate) is refining.Acquisition 0.41g (47%, 0.89mmol) white foam shape thing.

[α] ²³D＝-35.36°(c＝0.51，CHCl ₃)。

¹HNMR(300 MHz，CDCl ₃)δ0.93(3H，t，J＝7.0Hz)，0.96(3H，d，J＝6.6Hz)，1.46(9H，s)，2.06-2.13(1H，m)，2.50(1H，t，J＝4.0Hz)，2.94(1H，d，J＝15.0Hz)，3.04(1H，dd，J＝4.4，7.3Hz)，3.43(1H，dd，J＝3.3，6.6)，3.78(3H，s)，3.80-3.86(1H，m)，3.86(3H，s)，4.24(1H，d，J＝15.0Hz)，4.94(1H，d，J＝8.1Hz)，7.15(1H，bs)。

C ₁₉H ₃₀N ₂O ₉The analytical calculation value of S: C, 49.34; H, 6.54; N, 6.06.Measured value: C, 49.33; H, 6.44; N, 6.05.

MS(ES)m/z 461.2[M-H] ^-。

Preparation example 8

(1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-4 '-methyl-valeryl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, use commercially available acquisition N-BOC-(L)-leucine monohydrate and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 4) is prepared.Reaction mixture refluxed is spent the night.Use PC-TLC, 4mm SiO ₂Rotor, (10% ethyl acetate/hexane-100% ethyl acetate) purifying.Acquisition 0.85g (94%, 1.78mmol) white foam shape thing.

[α] ²³D＝-46.15°(c＝1.04，CHCl ₃)。

¹HNMR(300MHz，CDCl ₃)δ0.92(3H，d，J＝6.2Hz)，0.95(3H，d，J＝6.6Hz)，1.47(9H，s)，1.42-1.47(1H，m)，1.63-1.67(1H，m)，2.46(1H，t，J＝3.7Hz)，2.87(1H，d，J＝15.0Hz)，3.04(1H，dd，J＝4.4，7.3Hz)，3.41(1H，dd，J＝3.7，7.0)，3.78(3H，s)，3.86(3H，s)，4.00-4.05(1H，m)，4.20(1H，d，J＝15.0Hz)，4.75(1H，d，J＝6.6Hz)，7.43(1H，bs)。

C ₂₀H ₃₂N ₂O ₉The analytical calculation value of S: C, 50.41; H, 6.77; N, 5.88.Measured value: C, 50.30; H, 6.82; N, 5.75.

MS(ES)m/z 475.2[M-H] ^-。

Preparation example 9

(1R, 4S, 5S, 6S)-and 4-(2 ' S, 6-pair-tert-butoxycarbonyl amino-hexanamido)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, use commercially available acquisition N-BOC-Lys (BOC)-OH and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 4) is prepared.Reaction mixture refluxed is spent the night.Use PC-TLC, 4mmSiO ₂Rotor, (10% ethyl acetate/hexane-100% ethyl acetate) purifying.Acquisition 1.04g (93%, 1.76mmol) white foam shape thing.

[α] ²³D＝-32.0°(c＝0.5，CHCl ₃)。

¹H NMR(300MHz，CDCl ₃)δ1.44(9H，s)，1.46(9H，s)，1.39-1.53(3H，m)，1.56-1.65(1H，m)，1.77-1.84(2H，m)，2.50(1H，t，J＝4.4Hz)，2.98-3.20(4H，m)，3.42(1H，dd，J＝3.7，7.0Hz)，3.76(3H，s)，3.86(3H，s)，4.01(1H，dd，J＝7.7，13.2Hz)，4.09-4.19(1H，m)，4.71(1H，t，J＝7.3Hz)，5.13(1H，bs)，7.59(1H，bs)。

C ₂₅H ₄₁N ₃O ₁₁The analytical calculation value of S: C, 50.75; H, 6.98; N, 7.10.Measured value: C, 50.36; H, 6.99; N, 6.87.

MS(ES)m/z 590.2 [M-H] ^-。

Preparation example 10

(1R, 4S, 5S, 6S)-and 4-[2 ' S-tert-butoxycarbonyl amino-4 '-(trityl-formamyl)-butyrylamino]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, use commercially available acquisition N-BOC-(L)-glutamine (Trt)-OH and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 4) is prepared.Reaction mixture refluxed is spent the night.Use PC-TLC, 4mm SiO ₂Rotor, (10% ethyl acetate/hexane-100% ethyl acetate) is refining.Obtain 0.53g (48%, 0.72 mmol) white foam shape thing.

[α] ²³D＝-8.0°(c＝0.50，MeOH)。

¹H NMR(300MHz，CDCl ₃)δ1.42(9H，s)，1.83-1.88(1H，m)，2.03-2.18(1H，m)，2.16(1H，t，J＝4.0Hz)，2.57-2.64(1H，m)，2.60(1H，d，J＝15.0Hz)，2.64-2.80(1H，m)，2.88(1H，dd，J＝4.4，7.3Hz)，3.26(1 H，dd，J＝4.0，7.0Hz)，3.47(3H，s)，3.76-3.90(1H，m)，3.81(3H，s)，4.05(1H，d，J＝15.0Hz)，5.47(1H，bs)，7.02(1H，bs)，7.20-7.35(15H，m)，8.68(1H，bs)。

C ₃₈H ₄₃N ₃O ₁₀The analytical calculation value of S: C, 62.20; H, 5.91; N, 5.73.Measured value: C, 61.83; H, 6.09; N, 5.57.

MS(ES)m/z 731.9[M-H] ^-。

C ₃₈H ₄₃N ₃O ₁₀S[M+Na] ⁺The HRMS calculated value, 756.2567.Measured value, 756.2585.

Preparation example 11

(1R, 4S, 5S, 6S)-4-[(1 '-tert-butoxycarbonyl-tetramethyleneimine-2 ' S-carboxyl)-amino]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, use commercially available acquisition Boc-(L)-proline(Pro) (0.61g, 2.9mmol) and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6 two is imitated dimethyl phthalate (0.5g, 1.9mmol, preparation example 4) and is prepared.With PC-TLC (ethyl acetate/hexane) purifying, obtain 0.87g (99.2%) title compound.

[α] ²³D＝-61.2(c＝0.49，CHCl ₃)。

¹H NMR(300MHz，CDCl ₃)δ1.51(9H，s)，1.75-1.98(3H，m)，2.35-2.5(2H，m)，2.84(1H，d，J＝14.7Hz)，2.9-3.03(1H，m)，3.25-3.4(1H，m)，3.3-4.1(2H，m)，3.76(3H，s)，3.86(3H，s)，4.14-4.31(2H，m)，8.66(1H，s)。MS(ES)m/z 459.2[M-I] ^-。

Preparation example 12

(1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-4 '-methyl sulfenyl-butyrylamino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, use commercially available acquisition Boc-(L)-methionine(Met) (0.71g, 2.9mmol) and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (0.5g, 1.9mmol, preparation example 4) is prepared.With PC-TLC (ethyl acetate/hexane) purifying, obtain 0.85g (90.5%) title compound.

[α] ²³D＝-20.4(c＝0.49，CHCl ₃)。

¹HNMR(300MHz，CDCl ₃)δ1.46(9H，s)，1.69(1H，s)，1.8-2.05(2H，m)，1.9-2.0(1H，m)，1.95-2.3(3H，bs)，2.0-2.2(1H，m)，2.4-2.8(2H，m)，2.48(1H，t，J＝4.0Hz)，2.58(1H，bs)，2.92(1H，d，J＝14.7Hz)，3.01(1H，dd，J＝4.4，7.0Hz)，3.42(1H，dd，J＝3.7，7.3Hz)，3.78(3H，s)，3.87(3H，s)，4.22-4.24(2H，m)，5.06(1H，d，J＝7.7Hz)，7.27(1H，s)。MS(ES)m/z 493.1[M-1] ^-。

Preparation example 13

(1R, 4S, 5S, 6S)-and 4-[2 ' S-tert-butoxycarbonyl amino-3 '-(1-tert-butoxycarbonyl-1H-indol-3-yl)-propionamido]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, use commercially available acquisition Boc-(L)-tryptophane (Boc) (1.1g, 2.8mmol) and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (0.5g, 1.9mmol, preparation example 4) is prepared.With PC-TLC (ethyl acetate/hexane) purifying, obtain 0.7g (56.7%) title compound.

¹HNMR(300MHz，CDCl ₃)δ1.44(9H，s)，1.67(9H，s)，2.37(1H，bs)，2.86(1H，d，J＝15.0Hz)，2.88(1H，t，J＝4.4Hz)，3.15(2H，d，J＝6.6Hz)，3.39(1H，dd，J＝3.7，7.0Hz)，3.73(3H，s)，3.83(3H，s)，4.18(1H，d，J＝14.7Hz)，4.37-4.44(1H，m)，5.01(1H，bd，J＝8.1Hz)，7.11(1H，bs)，7.25-7.59(4H，m)，8.14(1H，bd，J＝8.4Hz)。

[α] ²³D＝-19.6(c＝0.51，CHCl ₃)。

C ₃₀H ₃₉N ₃O ₁₁S1.0C ₄H ₈O ₂The analytical calculation value: C, 55.35; H, 6.42; N, 5.70.

Measured value: C, 54.98; H, 6.09; N, 6.07.

C ₃₀H ₃₉N ₃O ₁₁Na ₁The HRMS calculated value of S, 672.2203.Measured value, 672.2180.

Preparation example 14

(1R, 4S, 5S, 6S)-and 4-[2 ' S-tert-butoxycarbonyl amino-3 '-(4-tert.-butoxy carbonyl oxygen base-phenyl)-propionamido]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, use commercially available acquisition 2S-tert-butoxycarbonyl amino-3-(4-tert.-butoxy carbonyl oxygen base-phenyl)-propionic acid (1.1g, 2.9mmol) and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (0.5g, 1.9mmol, preparation example 4) is prepared.With PC-TLC (ethyl acetate/hexane) purifying, obtain 0.94g (79.0%) title compound.

[α] ²³D＝+4(c＝1.00，CH ₃OH)。

¹H NMR(300MHz，CDCl ₃)δ1.44(9H，s)，1.56(9H，s)，2.44(1H，t，J＝4.0Hz)，2.88(1H，d，J＝14.7Hz)，2.98(1H，dd，J＝4.4，7.3Hz)，3.04(2H，d，J＝7.3Hz)，3.38(1H，dd，J＝4.0，7.3Hz)，3.77(3H，s)，3.83(3H，s)，4.11(1H，d，J＝14.3Hz)，4.22-4.29(1H，app q，J＝7.3Hz)，4.92(1H，bd，J＝7.7Hz)，7.07(1H，bs)，7.1-7.26(4H，m)。

C ₂₈H ₃₈N ₂O ₁₂The HRMS calculated value of SNa, 649.2043.Measured value, 649.2001.

Preparation example 15

(1R, 4S, 5S, 6S)-and 4-(3 '-acetoxyl group-2 ' S-tert-butoxycarbonyl amino-propionyl) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dimethyl dicarboxylate

According to general method A, do not use DMAP, use 3-acetoxyl group-2S-(tert-butoxycarbonyl amino) propionic acid (0.25g, 1.0mmol, preparation example 44) and (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (0.2g, 0.8mmol, preparation example 4) is prepared.With PC-TLC (ethyl acetate/hexane) purifying, obtain 0.19g (48.2%) title compound.

[α] ²³D＝-24(c＝1.0，CH ₃OH)。

¹HNMR(300MHz，CDCl ₃)δ1.47(9H，s)，2.10(3H，s)，2.51(1H，t，J＝4.4Hz)，2.99-3.07(2H，m)，3.43(1H，dd，J＝4.0，7.3Hz)，3.77(3H，s)，3.86(3H，s)，4.14-4.40(4H，m)，5.29(1H，bd，J＝7.3 Hz)，7.64(1H，bs)。

C ₁₉H ₂₈N ₂O ₁₁The HRMS calculated value of SNa, 515.1312.Measured value, 515.1305.

Preparation example 16

(1R, 2S, 4R, 5R, 6R)-and 2-amino-4-fluorine dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

Under the room temperature, (26mL 356mmol) dropped to (1R, 2S with thionyl chloride with 20 fens clock times, 4R, 5R, 6R)-and 2-amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid (14.45g, 71.12mmol, United States Patent (USP) 5,958,960) and in the slurries that in the 202mL dehydrated alcohol, form.Slurries are heated to reflux and stirred 3 hours.Make it be cooled to room temperature and stir and spend the night.With the gained solution for vacuum concentration is residuum, then with the dilution of 136mL ethyl acetate, and rotating by hand with the aqueous sodium carbonate of 306mL 10% and to handle 15 minutes, and be 10 thereby make final pH.Separate each layer, and (1 * 136mL) washs water layer with ethyl acetate.(organic extract, filtration and vacuum concentration that 1 * 136mL) washing merging is got up are the title compound of white solid thereby obtain 17.07g (93%) with salt solution.

[α] ²³D＝+20.37°(c＝1.1，MeOH)。

m.p.＝64-66℃

¹H NMR(400MHz，CDCl ₃)δ1.28(3H，t，J＝7.3Hz)，1.31(3H，t，J＝6.8Hz)，1.34-1.45(1H，m)，1.85(2H，bs)，2.17-2.21(2H，m)，2.32-2.34(1H，m)，2.49(1H，dd，J＝7.8，14.1Hz)，4.24(2H，dq，J＝1.5，7.3Hz)，5.33-5.52(1H，m)。

C ₁₂H ₁₈FNO ₄The analytical calculation value: C, 55.59; H, 7.00; N, 5.40.Measured value: C, 55.29; H, 6.75; N, 5.45.

MS (ES) m/z measured value 260.3[M+H] ⁺

Preparation example 17

(1R, 2S, 4R, 5R, 6R)-and 2-[2 ' S-(tert-butoxycarbonyl amino) propionyl] amino-4-fluorine is two Ring [3.1.0] hexane-2, the 6-diethyl dicarboxylate

Under-22 ℃, with 15 fens clock times under nitrogen atmosphere, N-Boc-L-L-Ala (38.62g, 204mmol) drip N-methylmorpholine (22.44mL in the solution that in the 396mL methylene dichloride, forms, 204mmol), drip isobutyl chlorocarbonate (26.48mL subsequently again, 204mmol), so that temperature of reaction is no more than-18 ℃.Stirred the gained dilute slurry 30 minutes down at-20 ℃, this moment is again with the (1R in 40 fens clock time adding 247mL methylene dichloride, 2S, 4R, 5R, 6R)-2-amino-4-fluorine dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (49.46g, 191mmol, preparation example 16) solution, so that temperature of reaction is no more than-16 ℃.After adding, reactant is removed from cooling bath, and stirred at ambient temperature 70 minutes, this moment, temperature of reaction reached 15 ℃, and color becomes greenish orange look.With 408mL 1NHCl processing reaction thing and stirred 5 minutes, separate each layer then.Use saturated NaHCO ₃The aqueous solution (1 * 408mL) washing organic layer, drying (Na ₂SO ₄), filter, and vacuum concentration is a white foam shape thing (88.16g).

C ₂₀H ₃₁FN ₂O ₇0.1CH ₂Cl ₂The analytical calculation value: C, 55.00; H, 7.16; N, 6.38.

Measured value: C, 55.18; H, 7.18; N, 6.49.

MS (ES) m/z measured value 431.3[M+H] ⁺, 331.2[M+H-Boc] ⁺

Preparation example 18

(1R, 2S, 4R, 5R, 6R)-and 2-[2 ' S-(tert-butoxycarbonyl amino) propionyl] amino-4-fluorine is two Ring [3.1.0] hexane-2, the 6-dicarboxylic acid

Under the room temperature; to (1R; 2S; 4R; 5R, 6R)-2-[2 ' S-(tert-butoxycarbonyl amino) propionyl] amino-4-fluorine dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (88.16g; 191mmol, preparation example 17) adds 238mL (477mmol) 2N sodium hydroxide in the solution that in 238mL THF, forms.With this two-phase mixture vigorous stirring 2.5 hours, reacted homogeneous phase and carry out this moment under the room temperature.With 238mL t-butyl methyl ether diluted mixture thing, mix afterwards, and separate each layer.Advance-go on foot the dilution water layer with 238mL water, and remove by filter particulate matter.(42.9mL, 515mmol) treatment soln is 30 minutes, chooses wantonly to add title compound subsequently as crystal seed, and stirs 1 hour with concentrated hydrochloric acid.Filter the gained slurries, and water (2 * 100mL) washings, 45 ℃ of following vacuum-drying 40 hours, thus obtaining the 72.2g title compound, it is a white solid.A part of solid (69.5g) stirred with 490mL acetone made vaporific solution in 1 hour; Filter and (2 * 100mL) wash with acetone.With this filtrate vacuum concentration is white foam shape thing, and under 45 ℃ of vacuum further dry 16 hours, thereby obtain 61.8g (86%, calibration has 12%wt/wt acetone) title compound.This material just is used for embodiment 14-18 without characterizing.

Preparation example 19

(1S, 2S, 4S, 5R, 6R)-the amino dicyclo of 4-acetoxyl group-2-(tert-butoxycarbonyl) [3.1.0] is own Alkane-2, the 6-diethyl dicarboxylate

Under the nitrogen atmosphere, to (1S, 2S, 4S, 5R, 6R)-and 2-(tert-butoxycarbonyl) amino-4-hydroxy dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (1.50g, 4.20mmol, United States Patent (USP) 5958960), pyridine (0.365mL, 4.62mmol) and DMAP (0.513g, 4.20mmol) add in the solution that in methylene dichloride (40mL), forms diacetyl oxide (0.514mL, 5.0mmol).Stirred 16 hours under the room temperature, with the methylene dichloride dilution, and the aqueous citric acid solution of impouring 10% (50mL).Water (50mL) and salt solution (50mL) washing organic layer.Use MgSO ₄Dry, filtration, and vacuum concentration make title compound, and it is white solid (1.295g, 75%).

LCMS:m/z 400 [M+H] ⁺With m/z 300[M+H-CO ₂Bu] ⁺@RT1.39 minute.

Preparation example 20

(1S, 2S, 4S, 5R, 6R)-and amino dicyclo [3.1.0] hexane-2 of 4-acetoxyl group-2-, 6-dicarboxylic acid two Ethyl ester

With (1S, 2S, 4S, 5R, 6R)-and the amino dicyclo of 4-acetoxyl group-2-(tert-butoxycarbonyl) [3.1.0] hexane-2,6-diethyl dicarboxylate (1.25g, 3.13mmol, preparation example 19) is dissolved in the solution of 95%TFA and methylene dichloride (60mL) formation, and under the room temperature nitrogen atmosphere, stirred 5 minutes.Vacuum is removed the TFA/ methylene dichloride afterwards.Crude product is dissolved in NaHCO ₃(1.00g) in the suspension that forms with methylene dichloride (50mL), and stirred 30 minutes.Filter this suspension, (3 * 25mL) washings, and vacuum concentration obtain 916mg (98%) product, and it is a yellow oil with methylene dichloride.

LCMS:m/z 400[M+H] ⁺@R _T0.92 minute.

Preparation example 21

(1S, 2S, 4S, 5R, 6R)-4-acetoxyl group-2-[2 ' S-(tert.-butoxy) carbonylamino propionyl] Amino dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

Under the nitrogen atmosphere, to (1S, 2S, 4S, 5R, 6R)-amino dicyclo [3.1.0] hexane-2 of 4-acetoxyl group-2-, 6-diethyl dicarboxylate (0.80g, 1.94mmol, preparation example 20) and Boc-L-Ala (0.477g, 2.52mmol) order adds EDC (0.519g in the suspension that forms in anhydrous methylene chloride (50mL), 2.72mmol), HOBt (0.314g, 2.32mmol), the DMAP of catalytic amount (0.024g, 0.19mmol) and triethylamine (1.08mL, 7.76mmol).Stir about is after 15 minutes under the room temperature, and initial white suspension thing dissolves fully.Reactant was stirred 16 hours,, and use saturated NaHCO with the methylene dichloride dilution ₃The aqueous solution (50mL), the 1.0N HCl aqueous solution (3 * 20mL) and saturated brine solution (40mL) washing.Use MgSO ₄Dry organic layer also filters and vacuum-drying.As elutriant, by the crude amide of column chromatography purifying gained, obtain 638mg (75%) product with (4: 1) mixture of ethyl acetate and hexane, it is a white solid.

¹H NMR(CD ₃OD) ^*：δ5.05(1H，d，5.7Hz)，4.09(2H，q，7.3Hz)，4.02(2H，q，7.3Hz)，4.01-3.94(1H，m)，2.65-2.59(2H，m)，2.38(1H，d，14Hz)，2.10-2.03(3H，m)，1.91(3H，s)，1.78(1H，dd，5.9Hz，16Hz)，1.59(1H，brs)，1.34(9H，s)，1.16(6H，2xt，7.3Hz)；

^*The unobservable exchangeable protons of N.B.NMR=2.

LCMS:m/z 471[M+H] ⁺With m/z 371 [M+H-CO ₂ ^tBu] ⁺@R _T1.30 minute.R _f0.50 (80% ethyl acetate: heptane)

Preparation example 22

(1S, 2S, 4S, 5R, 6R)-and 2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

Will (1S, 2S, 4S, 5R, 6R)-and 2-tert-butoxycarbonyl amino-4-hydroxy dicyclo [3.1.0] hexane-2, the solution that 6-diethyl dicarboxylate (US 5958960 for 5.0g, 14.0mmol) forms in ethyl acetate is cooled to 0 ℃.In solution, blast anhydrous HCl (g) until saturated and stirred 0.5 hour.Reaction is warming up to room temperature and stirred 1 hour.Concentrated reaction mixture also makes it at ethyl acetate and H ₂Distribute between the O.Use NaHCO ₃(aq) handle water layer, and use ethyl acetate extraction.Use K ₂CO ₃Dry organism also concentrates, and obtains 2.2g (59.7%) white solid.

[α] ²³D＝-30.8(c＝0.52，CH ₃OH)。

¹HNMR(300 MHz，CDCl ₃)δ1.26(3H，t，J＝7.33Hz)，1.36(3H，t，7.33Hz)，1.60(1H，dd，J＝5.87，15.40Hz)，1.64(1H，t，J＝2.93Hz)，2.27(2H，bs)，2.15-2.29(3H，m)，3.93(1H，bs)，4.13(2H，q，J＝6.97Hz)，4，27-4.36(3H，m)。C ₁₂H ₁₉NO ₅The analytical calculation value: C, 56.02; H, 7.44; N, 5.44.Measured value: C,

55.75；H，7.36；N，5.40。

MS(ES)m/z 258.1[M+H] ⁺。

Preparation example 23

(1S, 2S, 4S, 5R, 6R)-2-(2 '-tert-butoxycarbonyl amino-kharophen)-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, use the Boc-glycine (458mg, 2.62mmol, Aldrich) and (1S, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (450mg, 1.75mmol, preparation example 22) is prepared.Purifying on 35g silicon-dioxide is the hexane/ethyl acetate wash-out of 50/50-20/80 with gradient.Yield: 560mg (77%).

¹HNMR(400MHz，CDCl ₃)δ1.28(3H，t，J＝7.3Hz)，1.29(3H，t，J＝7.3Hz)，1.48(9H，s)，1.57(2H，m)，2.21(1H，m)，2.41(1H，dd，J＝2.9，5.8Hz)，2.81(1H，d，J＝15.6Hz)，3.76(1H，d，J＝5.8Hz)，4.10-4.18(3H，m)，4.26(2H，q，J＝7.3Hz)，4.33(1H，m)，5.1 0(1H，bs)，6.86(1H，bs)。

C ₁₉H ₃₀N ₂O ₈The analytical calculation value: C, 55.06; H, 7.30; N, 6.76.Measured value: C, 55.24; H, 7.50; N, 6.76.

MS(ES)m/z 41 5.2[M+H] ⁺，437.2[M+Na] ⁺。

Preparation example 24

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-tert-butoxycarbonyl amino-3 '-methylbutyryl amino)-4-hydroxyl Base-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, use the Boc-L-Xie Ansuan (569mg, 2.62mmol, Sigma) and (1S, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (450mg, 1.75mmol, preparation example 22) is prepared.Purifying crude product on 35g silicon-dioxide; With gradient is the hexane/ethyl acetate wash-out of 70/30-20/80.Yield: 694mg (87%) white foam shape thing.

¹HNMR(400MHz，CDCl ₃)δ0.95(3H，d，J＝6.8Hz)，1.00(3H，d，J＝6.8Hz)，1.29(6H，t，J＝7.3Hz)，1.45(9H，s)，1.52-1.60(2H，m)，2.08(1H，m)，2.20(1H，m)，2.42(1H，m)，2.81(1H，d，J＝15.6Hz)，3.86(1H，dd，J＝6.3，8.8Hz)，4.14(2H，q，J＝7.3Hz)，4.2 1-4.32(4H，m)，5.00(1H，d，J＝8.3Hz)，6.63(1H，s)。

C ₂₂H ₃₆N ₂O ₈The analytical calculation value: C, 57.88; H, 7.95; N, 6.14.Measured value: C, 57.87; H, 8.03; N, 6.12.

MS(ES)m/z 457.2 [M+H] ⁺，479.2 [M+Na] ⁺。

Preparation example 25

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-tert-butoxycarbonyl amino-4 '-methylpent amido)-4-hydroxyl Base-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, use the Boc-L-leucine (606mg, 2.62mmol, Chemlog) and (1S, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (450mg, 1.75mmol, preparation example 22) is prepared.Purifying crude product on 35 g silicon-dioxide; With gradient is the hexane/ethyl acetate wash-out of 70/30-20/80.Yield: 689mg (84%) white foam shape thing.

¹HNMR(400 MHz，CDCl ₃)δ0.96(6H，m)，1.28(3H，t，J＝7.3Hz)，1.29(3H，t，J＝7.3Hz)，1.46(9H，s)，1.45-1.73(5H，m)，2.20(lH，m)，2.42(1H，m)，2.80(1H，d，J＝16.1Hz)，4.07-4.35(7H，m)，4.81(1H，bd)，6.86(1H，bs)。

C ₂₃H ₃₈N ₂O ₈0.1H2O the analytical calculation value: C, 58.48; H, 8.15; N, 5.93.Measured value: C, 58.22; H, 7.94; N, 5.92.

MS(ES)m/z 471.2[M+H] ⁺，493.2[M+Na] ⁺。

Preparation example 26

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-tert-butoxycarbonyl amino-3 ' S-methylpent amido)-4- Hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, use the Boc-L-Isoleucine (606mg, 2.62mmol, Aldrich) and (1S, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (450mg, 1.75mmol, preparation example 22) is prepared.Purifying crude product on 35 g silicon-dioxide is the hexane/ethyl acetate wash-out of 70/=30-20/80 with gradient.Yield: 731mg (89%).

¹HNMR(400MHz，CDCl ₃)δ0.92(3H，t，J＝7.3Hz)，0.97(3H，d，J＝6.8Hz)，1.17(1H，m)，1.29(6H，t，J＝6.8Hz)，1.46(9H，s)，1.53(1H，dd，J＝5.8，15.6Hz)，1.58(1H，t，J＝2.4Hz)，1.74(1H，m)，1.83(1H，m)，2.20(1H，m)，2.43(1H，m)，2.80(1H，d，J＝15.6Hz)，3.88(1H，dd，J＝7.3，8.8Hz)，4.12-4.32(6H，m)，4.98(1H，d，J＝7.3Hz)，6.60(1H，s)。

C ₂₃H ₃₈N ₂O ₈0.2H ₂The analytical calculation value of O: C, 58.25; H, 8.16; N, 5.91.

Measured value: C, 58.17; H, 8.11; N, 5.91.

MS(ES)m/z 471.2 [M+H] ⁺，493.2 [M+Na] ⁺。

Preparation example 27

(1S, 2S, 4S, 5R, 6R)-2-[2 '-(2-tert-butoxycarbonyl amino-kharophen)-acetyl ammonia Base)-and 4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, (474mg is 2.04mmol) with (1S, 2S to use Boc-Gly-Gly, 4S, 5R, 6R)-and 2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate hydrochloride (400mg, 1.36mmol, do not use alkaline purification in the preparation example 22) be prepared, but following exception is arranged.Do not use DMAP.Add 1 equivalent triethylamine.The purifying crude product is used eluent ethyl acetate on 35g silicon-dioxide.Yield: 517mg (81%).

[α] ²³D＝-18.18(c＝0.55，MeOH)。

¹HNMR(300MHz，CDCl ₃)δ1.26(3H，t，J＝6.8Hz)，1.27(1H，t，J＝7.3Hz)，1.28(3H，t，J＝6.8Hz)，1.46(9H，s)，1.59(1H，t，J＝3.4Hz)，1.62(1H，dd，J＝6.3，16.1Hz)，2.21(1H，dd，J＝2.9，5.8Hz)，2.46(1H，dd，J＝2.4，5.8Hz)，2.73(1H，d，J＝15.6Hz)，3.78-3.91(3H，m)，4.01-4.15(3H，m)，4.24(2H，q，J＝6.8Hz)，4.32(1H，d，J＝5.8Hz)，5.28(1H，b)，6.87(1H，bt，J＝4.9Hz)，7.39(1H，bs)。

C ₂₁H ₃₄N ₃O ₉HRMS (mass spectrum) calculated value, 472.2295.Measured value, 472.2303.

HPLC:16.755 minute.Post: symmetrical C18,3.5um, 4.6 * 150 mm. λ=230nM.

Flow velocity: 1mL/ minute.Gradient: 10%-50%ACN/ contains the water of 0.1%TFA, 25 minutes time.

Preparation example 28

(1S, 2S, 4S, 5R, 6R)-2-[2 '-(2S-tert-butoxycarbonyl amino-propionamido)-acetyl ammonia Base)-and 4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, (502mg is 2.04mmol) with (1S, 2S to use Boc-Ala-Gly, 4S, 5R, 6R)-and 2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate hydrochloride (400mg, 1.36mmol, do not use alkaline purification in the preparation example 22) be prepared, but following exception is arranged.Do not use DMAP.Add 1 equivalent triethylamine.The purifying crude product is used eluent ethyl acetate on 35g silicon-dioxide.Yield: 500mg (76%).

[α] ²³D＝-31.37(c＝0.55，MeOH)。

¹HNMR (300MHz, CDCl ₃) δ 1.27 (3H, t, J=6.8Hz), 1.28 (3H, t, J=6.8Hz), 1.42 (3H, d, J=6.8Hz), 1.45 (9H, s), 1.58 (1H, t, J=3.4Hz), 1.64 (1H, dd, J=5.8,15.6Hz), 2.23 (1H, dd, J=3.4,5.8Hz), 2.50 (1H, dd, J=2.9,6.3Hz), 2.69 (1H, d, J=15.6Hz), the ABq of doublet (2H, ν A=3.87, ν B=3.98, J _AB=17.1Hz, J _d=6.3Hz), 3.92 (1H, m), 4.07-4.16 (3H, m), 4.24 (2H, q, J=7.3Hz), 4.32 (1H, b), 5.08 (1H, b), 6.84 (1H, bt, J=4.88 Hz), 7.14 (1H, bs).

C ₂₂H ₃₆N ₃O ₉The HRMS calculated value, 486.2452.Measured value, 486.2451.

Preparation example 29

(1S, 2S, 4S, 5R, 6R)-2-(2 '-tert-butoxycarbonyl amino-3 '-phenyl-propionamido)-4-hydroxyl Base-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, use the Boc-L-phenylalanine (772mg, 2.91mmol) and (1S, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (480mg, 1.94mmol, preparation example 22) is prepared.The silica filler that concentrate crude product mixture, be dissolved in methylene dichloride, usefulness ethyl acetate/DCM (1: 1) passes through one section weak point fast, and vacuum concentration obtains yellow oil.Be further purified with silica gel chromatography,, obtain 852mg (87%) title compound with 30% ethyl acetate/hexane to 80% ethyl acetate/hexane wash-out.

[α] ²³D＝-23.66(c＝0.93，MeOH)。

¹HNMR(400 MHz，CD ₃0D)δ1.22-1.30(6H，m)，1.35(9H，s)，1.59(1H，t，J＝2.9Hz)，1.70(1H，dd，J＝5.5，15.4Hz)，2.06(1H，m)，2.40(1H，d，J＝15.4Hz)，2.63(1H，dd，J＝2.9，5.9Hz)，2.76(1H，dd，J＝9.2，13.9Hz)，4.08-4.33(6H，m)，7.20-7.29(5H，m)。

MS measured value 505.0[M+H] ⁺

C ₂₆H ₃₆N ₂O ₈The HRMS calculated value, 505.2550.Measured value, 505.2559.

Preparation example 30

(1S, 2S, 4S, 5R, 6R)-2-[2 '-tert-butoxycarbonyl amino-4 '-(trityl-carbamyl Base)-butyrylamino]-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

Except that not using the DMAP, according to general method A, (1.40g is 2.86mmol) with (1S with Boc-L-glutamine (trityl)-OH, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (490mg, 1.90mmol, preparation example 22) is prepared.Concentrate crude product mixture, be dissolved in methylene dichloride, with ethyl acetate the silica filler by one section weak point fast, and vacuum concentration obtains yellow oil.Be further purified with silica gel chromatography,, obtain 1.3g (94%) title compound with 30% ethyl acetate/hexane-100% ethyl acetate/hexane wash-out.

[α] ²³D＝-21.28(c＝0.94，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ 1.18-1.26(6H，m)，1.44(9H，s)，1.62-1.74(3H，m)，1.91-2.00(1H，m)，2.05(1H，m)，2.38-2.48(3H，m)，2.54(1H，m)，4.00-4.27(6H，m)，7.18-7.30(15H，m)。

MS measured value 728.2[M+H] ⁺

C ₄₁H ₄₉N ₃O ₉The HRMS calculated value, 728.3547.Measured value, 728.3533.

Preparation example 31

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S, 6 '-two tert-butoxycarbonyl amino-caproyl amino)-4-hydroxyl Base-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

Except that DMAP not being added the reaction mixture, according to general method A, (910mg is 2.63mmol) with (1S, 2S with Boc-L-Methionin (Boc)-OH, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (450mg, 1.75mmol, preparation example 22) be prepared.Concentrate crude product mixture, be dissolved in methylene dichloride, with ethyl acetate the silica filler by one section weak point fast, and vacuum concentration obtains yellow oil.Be further purified with silica gel chromatography,, obtain 800mg (78%) title compound with ethyl acetate/hexane (1: 1) wash-out.

[α] ²³D＝-30.19(c＝0.53，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ1.21-1.24(6H，m)，1.26-1.77(27H，m)，2.07(1H，m)，2.42(1H，d，J＝15.8Hz)，2.59(1H，dd，J＝2.6，5.5Hz)，3.03(1H，t，J＝6.2Hz)，3.99(1H，m)，4.07-4.28(5H，m)。

MS measured value 586.1[M+H] ⁺

C ₂₈H ₄₇N ₃O ₁₀The HRMS calculated value, 586.3340.Measured value, 586.3348.

Preparation example 32

(1S, 2S, 4S, 5R, 6R)-2-[2 '-tert-butoxycarbonyl amino-3 '-(trityl-carbamyl Base) propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

Except that DMAP not being added the reaction mixture, according to general method A, (1.35g is 2.84mmol) with (1 S, 2S with Boc-L-asparagine (trityl)-OH, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (476mg, 1.85mmol, preparation example 22) be prepared.Concentrate crude product mixture, be dissolved in methylene dichloride, with ethyl acetate the silica filler by one section weak point fast, and vacuum concentration obtains yellow oil.Be further purified with silica gel chromatography,, obtain 1.07g (81%) title compound with 20% ethyl acetate/hexane-50% ethyl acetate/hexane wash-out.

[α] ²³D＝-25.45(c＝0.55，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ1.19-1.26(6H，m)，1.45(9H，s)，1.61-1.69(2H，m)，2.05(1H，m)，2.44(1H，d，J＝15.4Hz)，2.51-2.72(3H，m)，4.02-4.21(4H，m)，4.25(1H，d，J＝5.5Hz)，4.36(1H，dd，J＝4.8，8.8Hz)，7.18-7.30(15H，m)。

MS measured value 714.1[M+H] ⁺

C ₄₀H ₄₇N ₃O ₉The HRMS calculated value, 714.3391.Measured value, 714.3380.

Preparation example 33

(1S, 2S, 4S, 5R, 6R)-2-[2 '-tert-butoxycarbonyl amino-3 '-(1 '-tert-butoxycarbonyl- 1H-indoles-3 '-yl)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, use Boc-L-tryptophane (Boc)-OH (1.18g, 2.91mmol) and (1S, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (520mg, 1.90mmol, preparation example 22) is prepared.Concentrate crude product mixture, be dissolved in methylene dichloride, with ethyl acetate the silica filler by one section weak point fast, and vacuum concentration obtains yellow oil.Be further purified with silica gel chromatography,, obtain 1.2g (92%) title compound with ethyl acetate/hexane DCM (1: 1) wash-out.

[α] ²³D＝-12.5(c＝0.96，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ1.18-1.27(6H，m)，1.34(9H，s)，1.62(IH，t，J＝2.9Hz)，1.67(9H，s)，1.74(IH，dd，J＝5.9，15.2Hz)，2.07(1H，m)，2.46(1H，d，J＝15.7Hz)，2.60(1H，d，J＝2.9，5.9Hz)，2.89(1H，dd，J＝9.3，15.0Hz)，3.16(1H，dd，J＝4.9，15.2Hz)，4.04-4.12(2H，m)，4.18-4.24(2H，m)，4.27(1H，d，J＝5.4Hz)，4.43(1H，dd，J＝5.4，9.3Hz)，7.26(2H，m)，7.52(1H，s)，7.64(1H，d，J＝7.3Hz)，8.10(1H，d，J＝8.3Hz)。

MS measured value 644.8[M+H] ⁺, 666.8[M+Na] ⁺

C ₃₃H ₄₄N ₃O ₁₀The HRMS calculated value, 666.3002.Measured value, 666.2988.

Preparation example 34

(1S, 2S, 4S, 5R, 6R)-and 2-(1 '-tert-butoxycarbonyl-tetramethyleneimine-2 ' S-carbonyl)-4-hydroxyl-two Ring [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, use the Boc-L-proline(Pro) and (1S, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (preparation example 22) is prepared.The thick material of purifying on the 110g of ISCO system silicon-dioxide is the eluent ethyl acetate of 80% ethyl acetate/hexane-100% with gradient, obtains 699mg (84%) title compound.

[α] ²³D＝-52.53(c＝0.99，MeOH)。

¹H NMR(400MHz，CD ₃OD)δ1.26(6H，m)，1.44(9H，s)，1.62(1H，m)，1.73(1H，dd，J＝5.4，15.3Hz)，1.84(1H，m)，1.97(1H，m)，2.07(4H，m)，2.41(1H，d，J＝15.3Hz)，2.50(1H，m)，2.59(1H，m)，4.12(2H，m)，4.20(1H，q，J＝7.4Hz)，4.27(1H，d，J＝5.4Hz)。

MS measured value 454.9[M+H] ⁺, 476.8 [M+Na] ⁺

C ₂₂H ₃₄N ₂O ₈The HRMS calculated value of Na, 477.2213. measured value, 477.2210.

Preparation example 35

(1S, 2S, 4S, 5R, 6R)-2-[2 ' S-tert-butoxycarbonyl amino-3 '-(4-tert.-butoxy carboxylic oxygen base- Phenyl)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, use 2S-tert-butoxycarbonyl amino-3-(4-tert.-butoxy carbonyl oxygen base-phenyl)-propionic acid and (1S, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (preparation example 22) prepares title compound.The thick material of purifying on the 110g of Isco system silicon-dioxide is the ethyl acetate/hexane wash-out of 60% ethyl acetate/hexane-90% with gradient, obtains 1.09g (91%) title compound.

[α] ²³D＝-12(c＝1.0，MeOH)。

¹HNMR(400 MHz，CD ₃OD)δ1.25(6H，m)，1.37(9H，s)，1.52(9H，s)，1.68(1H，dd，J＝5.4，15.3Hz)，2.06(1H，m)，2.38(1H，d，J＝15.3Hz)，2.64(1H，m)，2.79(1H，dd，J＝8.4，13.9Hz)，3.05(1H，dd，J＝5.4，13.9Hz)，4.20(6H，m)，7.04(2H，d，J＝8.4Hz)，7.26(2H，d，J＝8.4Hz)。

MS measured value 621.8[M+H] ⁺, 643.8[M+Na] ⁺

C ₃₁H ₄₄N ₂O ₁₁The HRMS calculated value of Na, 643.2843.Measured value, 643.2845.

Preparation example 36

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-tert-butoxycarbonyl amino-4 '-methyl sulfenyl-butyryl radicals ammonia Base)-and 4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, use (Boc-L-methionine(Met)) and (1S, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (preparation example 22) prepares title compound.The thick material of purifying on the 110g of Isco system silicon-dioxide is the eluent ethyl acetate of 80% ethyl acetate/hexane-100% with gradient, obtains 1.0g (92%) title compound.

[α] ²³D＝-30(c＝1.0，MeOH)。

¹H NMR(400MHz，CD ₃OD)δ1.26(6H，m)，1.43(9H，s)，1.84(6H，m)，2.06(1H，m)，2.2(1H，m)，2.48(1H，m)，2.61(1H，m)，3.37(1H，m)，2.49(1H，m)，4.2(6H，m)。

MS measured value 488.8[M+H] ⁺, 510.8[M+Na] ⁺

C ₂₂H ₃₆N ₂O ₈The HRMS calculated value of SNa, 511.2090.Measured value, 511.2071.

Preparation example 37

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-tert-butoxycarbonyl amino-propionyl amino)-4-hydroxyl- Dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method A, use commercially available acquisition N-BOC-(L)-L-Ala and (1S, 2S, 4S, 5R, 6R)-2-amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (preparation example 22) is prepared.Reaction mixture refluxed is spent the night.With preparation HPLC, the SiO2 post of 1 * 500g, (10% ethyl acetate/hexane-100% ethyl acetate) purifying.Acquisition 3.0g (90%, 7.00mmol) white foam shape thing.

[α] ²³D＝-32.31°(c＝0.37，CHCl ₃)。

¹H NMR(300MHz，CDCl ₃)δ1.28(6H，t，J＝7.3Hz)，1.34(3H，d，J＝7.3Hz)，1.46(9H，s)，1.51-1.58(2H，m)，1.64(1H，s)，2.19(1H，dd，J＝3.3，6.2Hz)，2.40(1H，dd，J＝2.9，6.2Hz)，2.80(1H，d，J＝15.8Hz)，4.10-4.36(6H，m)，4.92(1H，bs)，6.94(1H，bs)。

C ₂₀H ₃₂N ₂O ₈0.1H ₂The analytical calculation value of O: C, 55.83; H, 7.54; N, 6.51.

Measured value: C, 55.57; H, 7.64; N, 6.44.

MS(ES)m/z 429.2[M+H] ⁺，329.1[M-Boc] ⁺。

Preparation example 38

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-amino-propionyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane -2,6-diethyl dicarboxylate hydrochloride

Under 0 ℃; with anhydrous HCl gas purging (1S; 2S; 4S, 5R, 6R)-2-(2 ' S-tert-butoxycarbonyl amino-propionyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (2.95g; 6.88mmol, preparation example 37) and the solution that forms in ethyl acetate (30mL) is saturated by HCl until this solution.0 ℃ is stirred the gained reaction mixture down, judges up to TLC to react completely.Purge reaction mixture 30 minutes to remove excessive HCl gas with N2.Under the vacuum gained suspension is concentrated into drying, obtain 2.47g (98%, the 6.78mmol) semicarbazide hydrochloride of Xi Wanging.Need not to be further purified.

[α] ²³D＝28.0°(c＝0.50，MeOH)。

¹HNMR(300MHz，CDCl ₃)δ1.24(3H，t，J＝7.0Hz)，1.28(3H，t，J＝7.3Hz)，1.50(3H，d，J＝7.3Hz)，1.61(1H，t，J＝2.9Hz)，1.76(1H，dd，J＝5.9，15.8Hz)，2.10(1H，dd，J＝3.3，5.9Hz)，2.43(1H，d，J＝15.4Hz)，2.60(1H，dd，J＝2.9，6.2Hz)，3.90(1H，q，J＝7.0，13.9Hz)，4.15(2H，q，J＝7.3Hz)，4.14-4.31(3H，m)。

C ₁₅H ₂₄N ₂O ₆HCl0.7H ₂The analytical calculation value of O: C, 47.73; H, 7.05; N, 7.42.Calculated value: C, 47.96; H, 6.91; N, 7.04.

MS(ES)m/z 329.1[M+H] ⁺。

Preparation example 39

(1S, 2S, 4S, 5R, 6R)-2-[2 ' S-(2S-tert-butoxycarbonyl amino-propionyl amino) propionyl Base is amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

Except that not using the DMAP; according to general method A; use (1S, 2S, 4S; 5R; 6R)-and 2-(2 ' S-amino-propionyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate hydrochloride (0.2g, 0.55mmol; preparation example 38) and Boc-(L)-L-Ala (0.16g 0.82mmol) is prepared.With PC-TLC (ethyl acetate/hexane) purifying, obtain 0.13g (47.3%) title compound.

[α] ²³D＝-46.2(c＝0.52，CHCl ₃)。

¹H NMR(300 MHz，CDCl ₃)δ1.27(3H，t，J＝7.0Hz)，1.28(3H，t，J＝7.0Hz)，1.36(3H，d，J＝7.0Hz)，1.37(3H，d，J＝7.0Hz)，1.44(9H，s)，1.58-1.65(2H，m)，2.19(1H，dd，J＝3.0，5.9Hz)，2.46(1H，dd，J＝2.6，5.9Hz)，2.70(1H，d，J＝15.4Hz)，4.09-4.33(7H，m)，4.48(1H，app p，J＝7.0Hz)，5.05(1H，bd，J＝6.6Hz)，6.79(1H，bd，J＝7.7Hz)，7.26(1H，s)。

C ₂₃H ₃₈N ₃O ₉The HRMS calculated value, 500.2608.Measured value, 500.2598.

Preparation example 40

(1S, 2S, 4S, 5R, 6R)-2-[2 ' S-(2-tert-butoxycarbonyl amino-acetylamino)-propionyl Base is amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

Except that not using the DMAP; according to general method A; (0.29g is 1.6mmol) with (1S, 2S to use the Boc-glycine; 4S; 5R, 6R)-2-(2 ' S-amino-propionyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate hydrochloride (0.4g; 1.1mmol, preparation example 38) be prepared.With PC-TLC (ethyl acetate/hexane) purifying, obtain 0.14g (26.2%) title compound.

[α] ²³D＝-14(c＝1.00，CDCl ₃)。

¹H NMR(300 MHz，CDCl ₃)δ1.27(3H，t，J＝7.3Hz)，1.29(3H，t，J＝7.3Hz)，1.38(3H，d，J＝7.0Hz)，1.45(9H，s)，1.57-1.65(2H，m)，2.19(1H，dd，J＝3.3，5.9Hz)，2.44(1H，dd，J＝2.9，5.9Hz)，2.74(1H，d，J＝15.8Hz)3.70-3.86(2H，m)，4.08-4.34(6H，m)，4.56(1H，app p，J＝7.0Hz)，5.31(1H，bs)，6.88(1H，bd，J＝7.0Hz)，7.50(1H，s)。

C ₂₂H ₃₆N ₃O ₉The HRMS calculated value, 486.2452.Measured value, 486.2444.

Preparation example 41

(1S, 2S, 4S, 5R, 6R)-2-[2 '-(2S-tert-butoxycarbonyl amino-4-methyl-pentanoyl ammonia Base)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method B; use N-BOC-(the L)-leucine monohydrate and the (1S of commercially available acquisition; 2S; 4S; 5R, 6R)-2-(2 ' S-amino-propionyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate hydrochloride (0.2g; 0.55mmol, preparation example 38) be prepared.Use PC-TLC, the SiO of 4mm ₂Rotor, (10% ethyl acetate/hexane-100% ethyl acetate) purifying.Acquisition 0.54g (62%, 1.00mmol) white foam shape thing.

[α] ²³D＝-61.2°(c＝0.49，MeOH)。

¹HNMR(300 MHz，CDCl ₃)δ0.93(3H，d，J＝6.2Hz)，0.94(3H，d，J＝6.2Hz)，1.25(3H，t，J＝7.3Hz)，1.27(3H，t，J＝7.3Hz)，1.36(3H，d，J＝7.0Hz)，1.43(9H，s)，1.53(1H，d，J＝9.5Hz)，1.55-1.69(5H，m)，2.18(1H，dd，J＝2.9，5.9Hz)，2.44(1H，dd，J＝2.9，6.2Hz)，2.70(1H，d，J＝15.8Hz)，4.06(1H.bs)，4.13(4H，q，J＝7.3Hz)，4.18-4.28(2H，m)，4.31(1H，d，J＝5.9Hz)，4.41-4.45(1H，m)，4.85(1H，bs)，6.57(1H，d，J＝7.3Hz)，6.97(1H，bs)。

C ₂₆H ₄₃N ₃O ₉0.1H ₂The analytical calculation value of O: C, 57.46; H, 8.01; N, 7.73.

Measured value: C, 57.18; H, 8.00; N, 7.64.

C ₂₆H ₄₃N ₃O ₉[M+Na] ⁺The HRMS calculated value, 564.2897.Calculated value, 564.2922.

Preparation example 42

(1S, 2S, 4S, 5R, 6R)-2-[2 '-(2S-tert-butoxycarbonyl amino-3-methyl-butyryl radicals ammonia Base)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

According to general method B, use N-BOC-(L)-Xie Ansuan and (1S, the 2S of commercially available acquisition; 4S, 5R, 6R)-2-(2 ' S-amino-propionyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate hydrochloride (0.2g, 0.55mmol, preparation example 38) is prepared.Use PC-TLC, the SiO of 4mm ₂Rotor, (10% ethyl acetate/hexane-67% ethyl acetate) purifying.Acquisition 0.36g (43%, 0.68mmol) white foam shape thing.

[α] ²³D＝-65.5°(c＝0.58，MeOH)。

¹H NMR(300MHz，CDCl ₃)δ 0.91(3H，d，J＝7.0Hz)，0.97(3H，d，J＝6.6Hz)，1.27(3H，t，J＝7.3Hz)，1.28(3H，t，J＝7.0Hz)，1.37(3H，d，J＝17.3Hz)，1.44(9H，s)，1.54-1.61(3H，m)，2.12-2.20(2H，m)，2.42(1H，dd，J＝2.6，5.9Hz)，2.70(1H，d，J＝15.4Hz)，3.90(1H，t，J＝6.6Hz)，4.13(2H，q，J＝7.0Hz)，4.20-4.29(2H，m)，4.31(1H，d，J＝5.9Hz)，4.40-4.48(1H，m)，4.93(1H，bs)，6.41(1H，d，J＝7.3Hz)，6.91(1H，bs)。

C ₂₅H ₄₁N ₃O ₉0.2CH ₂Cl ₂The analytical calculation value: C, 55.58; H, 7.66; N, 7.72.

Measured value: C, 55.76; H, 7.27; N, 7.49.

MS(ES)m/z 528.3 [M+H] ⁺。

Preparation example 43

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-acetylaminohydroxyphenylarsonic acid propionyl amino)-4-hydroxyl-dicyclo [3.1.0] Hexane-2, the 6-diethyl dicarboxylate

Under 0 ℃, stir (1S, 2S, 4S, 5R, 6R)-and 2-(2 ' S-amino-propionyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate hydrochloride (0.30g, 0.82mmol, preparation example 38) is at CH ₂Cl ₂The solution that forms (30mL), add earlier in order simultaneously triethylamine (0.16g, 1.6mmol), be then Acetyl Chloride 98Min. (0.10g, 1.18mmol).Reaction mixture stirred spend the night and allow its intensification,, and use NaHSO with ethyl acetate (700mL) diluted reaction mixture ₄The aqueous solution and salt water washing.With the dried over mgso organic layer.Use PC-TLC, the SiO of 4mm ₂The rotor purifying, (10% ethyl acetate/hexane-100% ethyl acetate), obtain 0.24g (79%, 0.65mmol) white foam shape thing.

[α] ²³D＝-48°(c＝0.5，MeOH)。

¹H NMR(300 MHz，CDCl ₃)δ1.27(3H，t，J＝7.3Hz)，1.28(3H，t，J＝7.3Hz)，1.37(3H，d，J＝7.0Hz)，1.55-1.62(2H，m)，1.99(3H，s)，2.19(1H，dd，J＝3.3，6.6Hz)，2.40(1H，dd，J＝2.6，5.9Hz)，2.77(1H，d，J＝15.8Hz)，4.07(2H，q，J＝7.0Hz)，4.21-4.31(2H，m)，4.33(1H，d，J＝6.2Hz)，4.43-4.47(1H，m)，6.13(1H，bs)，7.06(1H，bs)。

C ₁₇H ₂₆N ₂O ₇The analytical calculation value: C, 55.13; H, 7.08; N, 7.56.Calculated value: C, 55.05; H, 7.12; N, 7.29.

C ₁₇H ₂₆N ₂O ₇[M+Na] ⁺The HRMS calculated value, 393.1638.Measured value, 393.1644.

Preparation example 44

3-acetoxyl group-2S-(tert-butoxycarbonyl amino) propionic acid

(2.8g, 27.2mmol) (5.8g, (3.7g is 25.2mmol) at 1: 1 dioxane: in the solution that forms in the water 25.2mmol) to add the O-acetyl-L-serine with the heavy carbonic di tert butyl carbonate with N-methylmorpholine.Stirring reaction 24 hours makes it distribute between ethyl acetate and water then.With the ethyl acetate extraction aqueous solution and discard organism.Use NaHSO ₄The aqueous solution with pH regulator to 0-1.Ethyl acetate extraction, Na ₂SO ₄Drying, filtration and vacuum concentration.Get product 2.6g (41.7%) through the flash chromatography purifying.The gained material promptly is used for preparation example 15 without characterizing.

Preparation example 45

(1S, 5R)-3-(tert-butoxycarbonyl amino)-3-(tert-butoxycarbonyl)-6-oxabicyclo [3.1.0] Hexane

In 30 minutes with the Methylpropanedioic acid tert-butyl ester among the THF (385mL) (129g, 0.75mmol) solution add LiH among the THF (900mL) (14.9g, 1.875mol) and N, N-dimethyl allylidene urea (DMPU, 155g, 1.2mol) in the slurries of Xing Chenging, and the maintenance temperature is 0-5 ℃.Reaction mixture is heated to 65 ℃, and in 5.5 hours, adds suitable-1 among the THF (100mL), 4-two chloro-2-butylene (95%, 100g, 0.8mol, 1.08 equivalents) solution, keeping temperature is 63-67 ℃.Under 65 ℃ reaction mixture was stirred 4 hours.Reaction mixture is handled to such an extent that 1-(methoxycarbonyl)-1-(tert-butoxycarbonyl) encircles penta-3-alkene through water/MTBE.

¹H NMR(300MHz，CDCl ₃)δ5.57(s，2H，CH＝CH)，3.71(s，3H，CH ₃)，2.95(s，4H，2CH ₂)，1.42(s，9H，C(CH ₃) ₃)。

IR (film) 1734 (C=O), 1646,1268,1149cm ^-1

The reaction soln that will contain 1-(methoxycarbonyl)-1-(tert-butoxycarbonyl) ring penta-3-alkene is cooled to 10-15 ℃, and (1.3L is in cooling solution 1.3mol) (10-15 ℃) in 30 minutes it to be joined 1N NaOH.25 ℃ of following stirring reaction solution were also used the GC test monitoring in 24 hours.Hydrolysis reaction fully after, MTBE (645mL) joined in the reaction mixture and with solution stirring 5 minutes.Leave standstill each layer and separation.Abandon organic layer.Add 1.5M NaHSO ₄Solution (1470mL) is so that water layer acidifying (pH2-3).Add MTBE (1.3L) and separate each layer.With MTBE (385mL) aqueous layer extracted, and the organic layer that combines with the 5%LiCl solution washing.The vacuum concentration organic layer is also used heptane (780mL) dilution.Solution concentration to about 500mL, was stirred the gained slurries 1 hour under the room temperature.Cross filter solid, use heptane (250mL) washing and 35 ℃ of following vacuum-dryings, obtain 103.29g (61% yield) 1-(carboxylic acid)-1-(tert-butoxycarbonyl) ring penta-3-alkene, it is a white solid.

mp＝119℃。

¹HNMR(300MHz，CDCl ₃)δ5.61(s，2H，CH＝CH)，3.00(s，4H，2CH ₂)，1.46(s，9H，C(CH ₃) ₃)。

IR：3800-3000(br，COOH)，1741(CO ₂R)，1705(CO ₂H)，1283)，1149cm ^-1。

Under the nitrogen atmosphere, in having the 1L flask of mechanical stirrer, with 1-(carboxylic acid)-1-(tert-butoxycarbonyl) ring penta-3-alkene (50g, 236mmol) be dissolved in 850mL70: 30 toluene: among the MTBE, with thionyl chloride (33.6g, 283mmol, 1.2 equivalents) join in the reaction mixture that is stirred, keeping temperature is 23 ℃.Reaction soln is cooled to 0-5 ℃ and dripped triethylamine (32.2g, 318mmol, 1.35 equivalents) in 30 minutes.Reaction mixture is warming up to 23 ℃ and stirred 1 hour.Reaction mixture is dropped in the deionized water (625mL) fast, and keeping temperature is 20-25 ℃.Separate each layer and also use the NaHCO of 500mL 1M ₃The solution washing organic layer.Concentrate organic layer and separate 1-(chloroformyl)-1-(tert-butoxycarbonyl) ring penta-3-alkene, it is a weak yellow liquid.

¹HNMR(300 MHz，CDCl ₃)δ5.61(s，2H，CH＝CH)，3.04(app q，J＝15.1Hz，4H，2CH ₂)，1.49(s，9H，C(CH ₃) ₃)。

IR (film) 1796 (COCl), 1743 (CO ₂R), 1274,1233,1158cm ^-1

With the TBuA hydrosulfate in the deionized water (700mL) (0.81g, 2.4mmol) solution add sodiumazide (20.16g, 310mmol) in.In 45 minutes, the solution that contains 1-(chloroformyl)-1-(tert-butoxycarbonyl) ring penta-3-alkene in the MTBE/ toluene is added in the nitrine solution.23 ℃ of following stirred reaction mixtures 3 hours are analyzed until the GC of reaction soln and to be determined that 1-(chloroformyl)-1-(tert-butoxycarbonyl) ring penta-3-alkene is consumed.Separate each layer and use 1MNaHCO ₃(540mL) and deionized water (540mL, 270mL then) washing organic layer.Use NaSO ₄Dry organic layer also filters.Vacuum concentration obtains 1-(acid azide)-1-(tert-butoxycarbonyl) ring penta-3-alkene, and it is an oily matter.

¹H NMR(300MHz，CDCl ₃)δ5.58(s，2H，CH＝CH)，2.96(app t，J＝2.3Hz，4H，2CH ₂)，1.46(s，9H，C(CH ₃) ₃)。

IR (film) 2137 (CON ₃), 1720 (CO ₂R), 1246 (s, 1185,1136cm ^-1).

Under 95 ℃, in 1 hour, 1-(acid azide)-1-(tert-butoxycarbonyl) ring penta-3-alkene is joined in the 110mL toluene.The speed control of the nitrogen of emitting with this understanding to drip.In adition process, from reaction, distill out MTBE.95 ℃ of following stirring reactions 1 hour, and make its cool overnight to 23 ℃.The vacuum concentration solvent obtains 1-(isocyanate group)-1-(tert-butoxycarbonyl) ring penta-3-alkene, and it is an oily matter.

¹HNMR(300 MHz，CDCl ₃)δ5.67(s，2H，CH＝CH)，3.01(d，J＝15.6Hz，2H)，2.61(d，J＝15.6Hz，2H)，1.50(s，9H，C(CH ₃) ₃)。

IR (film) 2258 (NCO), 1732 (CO ₂R), 1157cm ^-1

Under the nitrogen atmosphere with the trimethyl carbinol (35g, 471mmol) add potassium tert.-butoxide (1M, among the THF, 471mL, 471mmol).Reaction soln is cooled to 0-5 ℃, adds the toluene solution that contains 1-(isocyanate group)-1-(tert-butoxycarbonyl) ring penta-3-alkene in 60 minutes, keeping temperature is 0-10 ℃.Make reaction be warming up to 23 ℃, stirred 2 hours, measure the disappearance of isocyanic ester initial substance with GC.Under 15 ℃, reaction mixture is added in the mixture of deionized water (1.2L) and MTBE (1.2L).Stirred solution 20 minutes also separates each layer.Through distilling to concentrate organic layer is become about 250mL.Add heptane (500mL), and overall solution volume is concentrated into 250mL.Gained pulpous state solution is cooled to 0 ℃, stirred 2 hours, filter.With cold heptane (2 * 100mL) washing leaching cakes and vacuum-drying, obtain 34.54g (52%, by 1-(carboxylic acid)-1-(tert-butoxycarbonyl) ring penta-3-alkene calculated yield) 1-(tert-butoxycarbonyl amino)-1-(tert-butoxycarbonyl) ring penta-3-alkene, it is a white solid.

m.p.87-89℃。

¹HNMR(500 MHz，CDCl ₃)δ5.63(s，2H，CH＝CH)，5.1(bs， ¹H，NH)，2.99(d，J＝17.2Hz，2H)，2.57(d，J＝16.0Hz，2H)，1.46(s，9H)，1.44(s，9H)。

¹³C NMR(CDCl ₃)δ173.3，154.9，127.7，81.1，64.5，44.8，28.3，27.8。IR(KBr)：3451，2981，2932，1712，1489，1369，1154cm ^-1。

MS (FIA) m/e (% relative intensity) 284.2 (M ⁺+ 1,56), 228.2 (73), 172.1 (97), 128.0 (100).

With the 1-in the 100mL ethyl acetate (tert-butoxycarbonyl amino)-1-(tert-butoxycarbonyl) ring penta-3-alkene (20.00g, 71mmol) solution adds TBuA hydrosulfate (4.08,10mmol, 0.17 monoperoxyphthalic acid magnesium hydrate (MMPA equivalent),, 52.4g, 4.76% active oxygen=2.1 equivalents) and the mixture of deionized water (100mL) in.With mechanical stirrer stirring reaction 19 hours.In 30 minutes, the sodium sulfite solution in the deionized water (100L) is added in the reaction mixture.With 100mL ethyl acetate diluted mixture thing and separate each layer.Spend IONS OF H ₂O (100mL), 2N NaOH (2 * 100mL) and deionization H ₂O (2 * 100mL) washing organic layers.Under 75-90 ℃ normal atmosphere, distill organic layer, reach 40mL until volume.Solution is cooled to 75 ℃ and add heptane (120mL).It is crystal seed that solution is cooled to 65 ℃ of also optional products with hope.Make the gained slurries be cooled to room temperature, in ice bath, cooled off 1 hour then.Filtration product is washed with 4: 1 cold heptane of 80mL: EtOAc, afterwards 40 ℃ of following vacuum-dryings, gets 15.45g (73% yield) target product, is white solid.Compound can be further from 2: 1 Virahol: recrystallization purifying the water.

m.p.130-133℃

¹HNMR(500MHz，CDCl ₃)δ5.0(bs，1H)，3.58(s，2H)，2.41(d，J＝15.3Hz，2H)，2.20(d，J＝15.2Hz，2H)，1.43(s，9H)，1.40(s，9H)。

¹³C NMR(CDCl ₃)δ171.3，154.3，81.2，79.5，62.8，57.0，38.6，28.3，27.7。

IR(KBr)：3453，2982，2932，1726，1708，1489，1369，1293，1156，840cm ^-1。

MS (FLA) m/e (% relative intensity) 300.3 (M ⁺+ 1,65), 244.4 (68), 188.2 (100) 144.1 (99).

Preparation example 46

(1S, 3R)-1,3-two (methyl-benzyl amino) propane dihydrochloride

Under the nitrogen atmosphere, (98%ee, 121g 1mol) are heated to 100 ℃ with (R)-α-Jia Jibianji amine.With dripped in 70 minutes dibromopropane (25.4mL, 50.5g, 250mmol).Mixture was heated 3 hours in addition, be cooled to 80 ℃ afterwards.With 10 minutes dropping dense (50%) NaOH solution.Add water (30mL) dissolved solids, in 30 minutes mixture is cooled to room temperature.Add MTBE (100mL).Add 100mL water dissolution throw out, and separate each layer.With 50mL salt water washing organic layer, drying (Na ₂SO ₄) and vacuum concentration, obtain the faint yellow oily thing of 126.0g.Under 70 ℃ (head temperature), use this oily matter of pillar vacuum distilling of 12-inch, to remove excessive (R)-α-Jia Jibianji amine.In having the 2L flask of agitator, residuum at the bottom of the still (the thick diamines of 68g) is dissolved in the 1L Virahol.With dripped in 10 minutes dense HCl (12M, 45mL, 540mmol).Add the 100mL Virahol in addition, mix fully to guarantee underflow liquid.Mixture was stirred 90 minutes and filtered.Use the washed with isopropyl alcohol filter cake, 40 ℃ of following vacuum-dryings obtain 64.32g (72% yield is based on dibromopropane) title compound, are white solid.

¹HNMR(500MHz，DMSO-d ₆)δ9.93(bs，2H)，9.47(bs，2H)，7.56(d，J＝7.0Hz，4H)，7.37(m，6H)，4.27(m，2H)，2.84(m，2H)2.48(m，2H)，2.1(m，2H)，1.56(d，J＝6.7Hz，6H)。

Preparation example 47

(1S)-1-(tert-butoxycarbonyl amino)-1-(tert-butoxycarbonyl)-ring penta-4-alkene-3-ketone

While stirring with (1R, 3R)-1,3-two (methyl-benzyl amino) propane dihydrochloride (24.65g, 69.3mmol, preparation example 46) is dissolved in the 150mL water.In solution, add 35mL (175mmol) 5N NaOH solution, add 150mL MTBE subsequently again.Stir after 15 minutes and to separate each layer, and with 100mL MTBE aqueous layer extracted.With the organic layer that the water washing of 100mL salt merges, use Na ₂SO ₄Dry also vacuum concentration obtains 18.06g (92% yield) 1R, and 3R-two (methyl-benzyl amino) propane is colorless oil.

In having the 500mL four-hole boiling flask of overhead, at N ₂Under the air-flow, with the 1R of 20.59g (73.5mmol), 3R-1,3-two (methyl-benzyl amino) propane is dissolved in respectively among 30mL exsiccant MTBE and the THF.Solution is cooled to-45 ℃ and in 17 minutes Dropwise 5 9mL (147mmol) n-butyllithium solution (2.5M is in the hexane).-45 ℃ were stirred yellow solution 2 hours down.With 28 fens clock times will (1S, 3S, 5R)-3-(tert-butoxycarbonyl amino)-3-(tert-butoxycarbonyl)-6-oxabicyclo [3.1.0] hexane (10.00g, 33.4mmol) add, and keeps temperature to be lower than-40 ℃ by the drips of solution in 55mL exsiccant THF.Funnel with extra 5mL THF rinsing dropping.-45 ℃ of following stirred reaction mixtures 18 hours are afterwards by dripping aqueous sulfuric acid (75mL) the cancellation reaction of 4N.Remove cooling bath after adding acid.Separate each layer, and with 50mL MTBE aqueous layer extracted.Use 50mL water and 50mL salt water washing blended organic layer respectively, use Na ₂SO ₄Drying, and vacuum concentration, it is thick 578242 to obtain 9.78g, is white solid.This solid is dissolved among the hot MTBE of 33mL, and adds heptane (66mL) in batches.Make the solution of stirring be cooled to room temperature, under 0 ℃, stirred the mixture 1 hour afterwards.Filter this solid, with 2: 1 cold heptane of 2 * 10mL: MTBE washing, and, obtain 8.52g (85% yield) (1S 35 ℃ of following vacuum-dryings 4 hours, 3R)-and 1-(tert-butoxycarbonyl amino)-1-(tert-butoxycarbonyl)-3-hydroxycyclopent-4-alkene, be white solid.

m.p.111-112℃。

Chirality HPLC analyzes: 99.7%ee.

[α] _D＝+114(c 1，MeOH)。

¹HNMR(500 MHz，CDCl ₃)δ 6.1(bs，1H)，5.9(bs，1H)，5.55(d，J＝5.0Hz，1H)，4.8(m，1H)，4.44(d，J＝10.5Hz，1H)，2.87(dd，J＝14.5，7.5Hz，1H)，2.00(d，J＝14.5Hz，1H)，1.45(s，9H)，1.42(s，9H)。IR(KBr)：3413，2983，1703，1491，1370，1309，1255，1155，1055cm ^-1。MS (FIA) m/e (% relative intensity) 300.3 (M ⁺+ 1,15), 226.2 (29), 170.1 (100), 126.1 (89), 108.3 (20).

With 2,2,6,6 ,-tetramethyl--1-piperidines oxygen base (free radical) (TEMPO) (0.84,5.3mmol) and KBr (0.63g, 5.3mmol, in the 2mL water) add (1S, 3R)-(20g is 66.8mmol) in the solution that forms in MTBE (200mL) for 1-(tert-butoxycarbonyl amino)-1-(tert-butoxycarbonyl)-3-hydroxycyclopent-4-alkene.Reaction mixture is cooled to 0 ℃, drips and contain NaHCO ₃NaOCl (8.4g) (3.14%, 240g, 100mmol) solution keeps temperature to be lower than 5 ℃.0 ℃ of following stirring reaction 1 hour also is heated to room temperature.Separate each layer, with MTBE (2 * 200mL) aqueous layer extracted.With the organic layer that the 200mL 1NHCl solution washing that contains 2.21g KI merges, use 10%Na afterwards ₂SO ₃Solution (200mL) washing.Water (2 * 200mL) washing organic layers, and vacuum concentration is to dry.Under 50 ℃, crude product is dissolved among the MTBE (60mL), and adds heptane (200mL) crystallization 1 hour.In 2 hours mixture is cooled to 0 ℃, filters then.With the cold heptane of 100mL: MTBE (65: 35) washing leaching cake and vacuum-drying, 16.99g (89% yield) title compound, be white solid.

m.p.116-18℃。

[α] _D＝+123(c1，MeOH)。

¹H NMR(500MHz，CDCl ₃)δ7.4(bs，1H)，6.32(d，J＝5.5Hz，1H)，5.6(bs，1H)，2.87(d，J＝18.2Hz，1H)，2.9(d，J＝18.2Hz，1H)，1.43(s，18H)。

¹³C NMR(CDCl ₃)δ206.1，170.2，160.8，154.9，136.1，84.5，66.1，46.6，28.9，28.4。

IR(KBr)：3419，2983，1722，1487，1730，1300，1259，1151，1012cm ^-1。

MS (FIA) m/e (% relative intensity) 254.2 (M ⁺+ 1,11), 242.3 (18), 228.2 (13), 186.1 (76), 143.2 (11), 242.3 (100).

Preparation example 48

Bromination 1-(2-tert.-butoxy-2-oxoethyl) tetramethylene sulfide

In the 22L flask, divide clock time with 30-60, with the monobromo-acetic acid tert-butyl ester (2.44L, 16.52 mole, 1 equivalent) adding tetramethylene sulfide (2.19L, 24.8 moles, 1.5 equivalent) in the solution that in acetone (11.38L), forms, during to keep temperature with water-bath be 15-25 ℃.Stirring reaction 22 hours is used ¹H NMR analytical sample reacts completely determining.Filtering precipitate with acetone (2L) washing, and 28-33 ℃ of following vacuum-drying 3 days, gets 4.328Kg (92.5% yield) title compound. ¹H NMR(500MHz，DMSO d-6)δ4.40(s，1H)，3.51(m，2H)，3.48(m，2H)，2.23(m，2H)，2.13(M，2H)，1.42(s，9H)。

Preparation example 49

(1S, 2S, 5R, 6R)-and 2-(tert-butoxycarbonyl amino)-4-oxo-dicyclo [3.1.0] hexane-2,6- The di-tert-butyl dicarboxylate

Under the nitrogen atmosphere, with 10 fens clock times with KOtBu (42mL, 42mmol, 1M solution among the THF, 2.5 equivalent) join under 0 ℃, bromination 1-(2-tert.-butoxy-2-oxoethyl) tetramethylene sulfide (11.9g, 42mmol, 2.5 equivalents in the acetonitrile (30mL), preparation example 48) in the solution, keep temperature to be lower than 5 ℃.Stir this milk sap, cooled off 1.5 hours.Drip trifluoroethanol (6.9g, 69mmol, 4.1 equivalents).In 5 minutes, add (1S)-1-(tert-butoxycarbonyl amino)-1-(tert-butoxycarbonyl)-ring penta-4-alkene-3-ketone (5g, 16.8mmol, preparation example 47) and trifluoroethanol (13.3g, 132mmol, 7.9 equivalents) solution in acetonitrile (20mL), keeping temperature is 3-5 ℃.0-5 ℃ of following stirred solution 4.5 hours.In the refrigerative reaction mixture, add MTBE (155mL) and H ₂O (80mL).Separate each layer, use H ₂O (50mL) washs organic layer, uses 20% salt solution (50mL) washing afterwards.Through the normal pressure distillation organic layer is concentrated into about 30mL.Add heptane (100mL) and concentrated solution.Adding heptane again reaches 93 ℃ until the vapor temperature of overhead product in addition on demand.Add THF (65mL), get target product solution.

Preparation example 50

(1S, 2S, 5R, 6R)-and 2-(tert-butoxycarbonyl amino)-4-oxo-dicyclo [3.1.0] hexane-2,6- The di-tert-butyl dicarboxylate

With the saturated K of 1.7L ₂CO ₃The aqueous solution join 0 ℃, 2.2L CH ₂Cl ₂In bromination 1-(2-tert.-butoxy-2-oxoethyl) tetramethylene sulfide (757g, 2.67mol, preparation example 48) solution in, keep temperature to be lower than 10 ℃.This two-phase mixture stirring after 1.5 hours, is added the NaOH solution of 223mL 50% in batches, keep temperature to be lower than 5 ℃.Stirred the mixture 3 hours and filtration.Use CH ₂Cl ₂This salt of rinsing.Separate each layer, and use 600mL CH ₂Cl ₂Aqueous layer extracted.Use solid K ₂CO ₃Dry organic layer and the vacuum concentration that merges gets 533.3g (98% yield) (2-tert.-butoxy-2-oxoethyl) tetramethylene sulfide, is light yellow oil.When refrigerator storage, this oily matter crystallization is a white solid.

mp 48-50℃。

¹H NMR(500 MHz，CDCl ₃)δ3.12(m，2H)，3.00(m，2H)，2.85(s，1H)，2.41(m，2H)，1.84(m，2H)，1.38(s，9H)。

Under 474mL (6.5mol, 10 equivalents) trifluoroethanol added 0 ℃, 650mL CH ₂Cl ₂In (1S)-1-(tert-butoxycarbonyl amino)-1-(tert-butoxycarbonyl)-ring penta-4-alkene-(194g is 653mmol) in the solution for 3-ketone.Drip 325mL CH with 40 fens clock times ₂Cl ₂In (2-tert.-butoxy-2-oxoethyl) tetramethylene sulfide (396g, 1.96mol) solution keep temperature to be lower than 10 ℃.Remove ice bath after 1 hour.Add deionized water (680mL) and separate each layer.Use 400mLCH ₂Cl ₂Aqueous layer extracted.With the organic layer that the water washing of 500mL salt merges, use Na ₂SO ₄Drying, and vacuum concentration get the faint yellow oily solid of 587g.Solid is dissolved in 400mLCH ₂Cl ₂In, with 5: 1: 1 hexane: methyl tertiary butyl ether: CH ₂Cl ₂Be elutriant, through the silica gel packed column wash-out of 1.6Kg, getting cumulative volume is the elutriant of 13.2L.Concentrate this elutriant, get the 398.7g white solid.Solid is dissolved in 70: 30 the hexane that 3L refluxes: among the MTBE.Solution is cooled to ambient temperature overnight, on ice bath, cooled off 1 hour then.Cross filter solid, with cold solvent (about 700mL) rinsing, 35 ℃ of following vacuum-dryings, must 173g (64% yield) (1S, 2S, 5R, 6R)-and 2-(tert-butoxycarbonyl amino)-4-oxo-dicyclo [3.1.0] hexane-2, the 6-di-tert-butyl dicarboxylate is white solid.

mp 144-46℃。

[α] _D＝+30.5(c1，CHCl ₃)。

¹H NMR(500MHz，CDCl ₃)δ5.36(d，1H)，2.88(m，1H)，2.64(dd，J＝5.2，3.2Hz，1H)，2.37(d，J＝2.7Hz，1H)，2.23(bs，1H)，1.45(s，9H)，1.43(s，18H)。

¹³C NMR(125 MHz，CDCl ₃)δ 206.2，171.2，168.5，155.3，83.4，82.7，80.7，61.2，43.2，36.0，34.3 28.4，28.2，28.0，25.3。

IR(CHCl ₃)：2982，1744，1719，1485，1394，1309cm ^-1。

MS (ES+) m/e (% relative intensity) 412.2 (M ⁺+ 1,79), 356.2 (50), 300.1 (97),

276.1(68)，244.1(100)。

C ₂₁H ₃₃NO ₇(411.29) analytical calculation value: C, 61.30; H, 8.08; N, 3.40.

Measured value: C, 61.32; H, 8.04; N, 3.51.

Preparation example 51

(1S, 2S, 4S, 5R, 6R)-2-(tert-butoxycarbonyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane- 2, the 6-di-tert-butyl dicarboxylate

Under the nitrogen atmosphere, the solution of preparation example 49 is cooled to 0 ℃, and drips 3-sec-butyl lithium borohydride (L-Selectride ^TM, 1M, among the THF, 21mL, 21mmol).0 ℃ of following stirred reaction mixture 45 minutes.Drip the sodium carbonate solution (31mL) of 2M, keep temperature to be lower than 8 ℃.Be added in the 30%H that forms in the 20mL water ₂O ₂(7.15g 63mmol), keeps temperature to be lower than 15 ℃ to solution.Stir after 10 minutes, add MTBE (210mL) and deionized water (100mL).Separate each layer, and use saturated Na ₂SO ₃Solution (40mL) and 1M NaHSO ₄Solution (40mL) washing organic layer.Under normal pressure, the distillation of organic layer volume is concentrated into about 90mL.Continue distillation and add heptane to keep constant volume.When the overhead product vapor temperature reaches 93 ℃, solution is cooled to 70 ℃ and add 7mL THF.Solution is cooled to 0 ℃ and stirred 1 hour.Cross filter solid, with cold heptane (10mL) washing, 50 ℃ of following vacuum-dryings, obtain 4.68g (67% yield) (1S, 2S, 4S, 5R, 6R)-2-(tert-butoxycarbonyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-di-tert-butyl dicarboxylate is white solid.

m.p.187-88℃。

¹H NMR(CDCl ₃，500MHz)δ 5.31(bs，1H)，4.38(d，J _AB＝10.5Hz，1H)，4.30(dd，J＝11.0，6.0Hz，1H)，2.68(d，J＝15.3Hz，1H)，2.17(m，1H)，2.07(m，1H)，1.58(m，J＝15.2Hz，1H)，1.45(s，9H)，1.44(s，9H)，1.43(s，9H)。

¹³C NMR(75MHz，CDCl ₃)δ175.2，170.5，155，83.2，81.3，80.1，73.7，66.9，43.1，36.0，34.4，29.3，28.3，28.1，27.4，22.1。

IR(CHCl ₃)：3445，2982，1714，1485，1361cm ^-1。

MS (ES+) m/e (% relative intensity) 414.2 (M ⁺+ 1,58), 358.1 (75), 302.1 (78), 246.0 (100).

[α] _D＝-28.6(c1，MeOH)。

Preparation example 52

(1R, 2S, 4R, 5R, 6R)-2-(2 ' S-2 '-(tert-butoxycarbonyl amino) propionyl) amino-4-fluoro- Dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

In 2 hours, with CH ₂Cl ₂(230mL) (1S, 2S, 4S, 5R, 6R)-2-(tert-butoxycarbonyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-di-tert-butyl dicarboxylate (25.00g, 60.5mmol, preparation example 51) solution join-78 ℃, CH ₂Cl ₂In Deoxofluor (105mL) (16.05g, 72.6mmol, the 1.2 equivalents) solution.Stirred reaction mixture 1.5 hours adds 1.66g Deoxofluor in addition again ^TMStirred solution 30 minutes also is warming up to-10 ℃.Splash into saturated NaHCO in 20 minutes ₃(105mL) solution keeps temperature to be lower than 5 ℃.Add the saturated NaHCO of 160mL ₃Solution is with the pH regulator to 7 of water layer.Mixture is warming up to room temperature and separates each layer.Use 100mL CH ₂Cl ₂Aqueous layer extracted.With the organic layer that the water washing of 250mL salt merges, use Na afterwards ₂SO ₄Dry.Remove under the vacuum and desolvate and heptane (75mL) is added in the crude product.With gained mixture heating up to 50 ℃, all dissolve until all solids, and at room temperature stirred 24 hours.In ice bath mixture being cooled to 0 ℃ also filters.With cold heptane rinsing product and vacuum-drying, obtain 20.23g (80% yield) (1R, 2S, 4R, 5R, 6R)-2-(tert-butoxycarbonyl amino)-4-fluoro-dicyclo [3.1.0] hexane-2, the 6-di-tert-butyl dicarboxylate is white solid.

m.p.140-143℃。

[α] _D＝+20.6(c1，CHCl ₃)。

¹HNMR(CDCl ₃，500MHz)δ 5.45(dd，J _H-F＝56，J _H-H＝4.8Hz，1H)，5.28(bs，1H)，3.00(m，1H)，2.23(bs，1H)，2.11(m，1H)，2.08(m，1H)，1.46(s，9H)，1.45(s，9H)，1.43(s，9H)，1.37(m，1H)。

¹³C NMR(CDCl ₃，75MHz)δ177.7，170.5，155.0，94.4，92.0，82.2，81.5，80.2，64.7，37.7，33.2，29.7，29.3，28.3，28.1，27.8，20.4。MS (ES+) m/e (% relative intensity) 416.2 (M ⁺+ 1,66), 360.1 (67), 304.1 (100), 248.0 (60).

IR(CHCl ₃)：3444，2981，1715，1485，1369cm ^-1。

Be not with under the refrigerative condition, with 10 minutes with SOCl ₂(21.29g, 13.05mL, 0.179 mole, 5 equivalents) and drop to (1R, 2S, 4R, 5R, 6R)-2-(tert-butoxycarbonyl amino)-4-fluoro-dicyclo [3.1.0] hexane-2,6-di-tert-butyl dicarboxylate (14.87g, 0.036 mole) (abs is 149mL) in the middle solution that forms, to prepare the solution of slow backflow at EtOH.The solution backflow is spent the night.From reaction, remove under the vacuum and desolvate.Residuum is dissolved among the EtOAc (150mL), and in 5-10 minute, drips 10% Na while stirring ₂CO ₃(75mL) solution.Separate each layer and use EtOAc (50mL) aqueous layer extracted.(organic extract that 1 * 50mL) washing merges is used Na with salt solution ₂SO ₄Drying is filtered and with product, (1R, 2S, 4R, 5R, 6R)-and 2-amino-4-fluoro-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate vacuum concentration is a thick liquids, it leaves standstill becomes solid (11.02g).

Under-22 ℃ the nitrogen atmosphere, with N-methylmorpholine (22.44mL, 204mmol) add to N-Boc-L-L-Ala (38.62g, 204mmol) in the solution that in the 396mL methylene dichloride, forms, afterwards with 15 minutes dropping isobutyl chlorocarbonate (26.48mL, 204mmol), thus make temperature of reaction be no more than-18 ℃.-20 ℃ were stirred the gained dilute slurry 30 minutes down, wherein add (1R with 40 fens clockwise then, 2S, 4R, 5R, 6R)-2-amino-4-fluoro-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (49.46g, 191mmol) the solution that in the 247mL methylene dichloride, forms, thus make temperature of reaction be no more than-16 ℃.Removing this reaction from cooling bath also at room temperature stirred 70 minutes.Add 408mL 1N hydrochloric acid, stirred 5 minutes, and separate each layer.With saturated sodium bicarbonate solution (1 * 408mL) washing organic layer, drying (Na ₂SO ₄), filter and with product, (1R, 2S, 4R, 5R, 6R)-2-(2 ' S-2 '-(tert-butoxycarbonyl amino) propionyl) amino-4-fluoro-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate vacuum concentration is a white foam shape thing (88.16g).

Under the room temperature; 46.7mL (93.4mmol) 2N sodium hydroxide is added to slightly (1R; 2S; 4R; 5R; 6R)-and 2-(2 ' S-2 '-(tert-butoxycarbonyl amino) propionyl) amino-4-fluoro-dicyclo [3.1.0] hexane-2, in the solution that 6-diethyl dicarboxylate (17.5g, 37.3mmol theoretical value) forms in the 46.6mL tetrahydrofuran (THF).This two-phase mixture of vigorous stirring is until mixing under the room temperature, and then stirs 1 hour (amounting to 3 hours).With 46mL t-butyl methyl ether diluted mixture thing, remix is 10 minutes afterwards, separates each layer then.In separating funnel, add 93mL water, add the dense HCl of 8.4mL (101mmol) again.Optional adding in this acidic solution (1R, 2S, 4R, 5R, 6R)-and 2-(2 ' S-2 '-(tert-butoxycarbonyl amino) propionyl) amino-4-fluoro-dicyclo [3.1.0] hexane-2,6-dicarboxylic acid crystal seed adds above-mentioned water layer afterwards again.Slowly add water layer earlier, so just form appropriate thick slurries.Improve again this moment and add speed (time of interpolation always is 40 minutes).Water (16mL) rinsing dropping funnel.Stir the gained slurries 2 hours, filtration, water (2 * 32mL) washings, and 45 ℃ of following vacuum-dryings to constant weight, obtain 13.9g (99%) title compound, be white solid.

Preparation example 53

(1R, 4S, 5S, 6S)-and 4-amino-2,2-dioxo-2 λ ⁶-thia-ring [3.1.0] hexane-4, the 6-diethyl dicarboxylate

Under the room temperature, with 20 fens clock times with thionyl chloride (15.5mL, 212.6mmol) drop to (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4 in the solution that 6-dicarboxylic acid (10g, 42.5mmol, United States Patent (USP) 5,688,826) forms, is used the rinsing of 40mL ethanol afterwards in 100mL 2B ethanol.Slurries are heated to reflux and stir and spend the night.Gained solution is cooled to room temperature and the gluey residuum of simmer down to.In residuum, add ethyl acetate (50mL), subsequently with other 94mL ethyl acetate dilution.Slowly the aqueous sodium carbonate (70mL) of adding 15% is so that it dissolves gradually in mixture while rotate by hand, and the gained final pH is 7.95.Filtration also separates each layer.With ethyl acetate (2 * 100mL) aqueous layer extracted.With salt solution (organic extract, drying (MgSO that 1 * 100mL) washing merges ₄), filter and vacuum concentration, obtain the title compound of faint yellow oily thing, it is cured as the solid (11.71g, 95% yield) of off-white look.

m.p.80-83℃。

[α] ²⁵D-57.7(c 1.04，CH ₃OH)。

500MHz ¹H NMR(CDCl ₃)δ 4.31(q，2H，J＝7.0Hz)，4.20(m，2H)，3.78(d，1H，J＝5.0Hz)，3.36(dd，1H，J＝4.0，7.0Hz)，2.93(dd，1H，J＝4.0，7.0Hz)，2.81(d，1H，J＝15.0Hz)，2.46(t，1H，J＝4.0)，1.34(t，3H，J＝7.0)，1.30(t，3H，J＝7.0)。

¹³C NMR(125MHz，CD ₃C1 ₃)δ 171.68，168.57，63.26，62.42，59.96，56.06，43.78，32.25，22.49，14.31，14.25。

FTIR(ATR)3364.15(s)，1725.95(s)，1304.91(s)，1259.24(s)，1200.84(s)，1104.91(s)，1022.99(s)，896.45(s)，851.21(s)cm ^-1。

C ₁₁H ₁₇NO ₆The analytical calculation value of S: C, 45.35; H, 5.88; N, 4.81.Measured value: C, 45.02; H, 5.75; N, 4.82.

Preparation example 54

(1R, 4S, 5S, 6S)-and 4-(2 ' S-4 '-methylthio group-2 '-(tert-butoxycarbonyl) amino butyryl radicals) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-diethyl dicarboxylate

Under-22 ℃ the nitrogen atmosphere, with N-methylmorpholine (14.4mL, 130.9mmol) add to N-Boc-L-methionine(Met) (32.64g, 130.9mmol) in the settled solution that in the 110mL methylene dichloride, forms, afterwards with 7 minutes dropping isobutyl chlorocarbonate (17mL, 130.9mmol), so that temperature of reaction remains on-22 ℃.Drip the dilute slurry that finishes then to form gained.-22--26 ℃ following the stirring 30 minutes.In 15 minutes, to wherein add (1R, 4S, 5S, 6S)-4-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the solution that 6-diethyl dicarboxylate (35.65g, 122.4mmol, preparation example 53) forms in the 107mL methylene dichloride is used the rinsing of 36mL methylene fluoride afterwards.Removing this reaction from cooling bath also at room temperature stirred 70 minutes.In solution, add 51mL 5N hydrochloric acid, separate each layer then.With methylene dichloride (2 * 107mL) reextraction water layers.With saturated sodium bicarbonate solution (organic layer, drying (MgSO that 2 * 107mL) washings merge ₄), filter and vacuum concentration, obtain 65.82g (103% weight yield) title compound, be white foam shape thing.

[α] ²⁵D-12.7(c 1.2，CH ₃OH)。

500 MHz ¹H NMR(CDCl ₃)δ7.53(s，1H)，5.06(d，1H，J＝8.0Hz)，4.34-4.20(m，6H)，3.41(dd，1H，J＝4.0，7.0)，2.97-2.89(m，2H)，2.64-2.59(m，2H，J＝4.0)，2.12-1.89(m，5H)，1.47(s，9H)，1.32(t，6H，J＝7.0)。

¹³C NMR(125MHz，CD ₃OD)δ 172.53，169.03，167.88，156.00，80.62，63.45，62.56，60.20，55.33，52.78，42.81，31.52，31.38，30.12，28.49，22.69，15.44，14.23，14.143。

FTIR(ATR)3341.88(w)，2979.38(s)，1733.03(s)，1674.92(s)，1514.58(s)，1315.80(s)，1255.15(s)，1161.47(s)，1142.63(s)，1025.68(s)，854.85(s)，763.53(s)cm ^-1。

Preparation example 55

(1R, 4S, 5S, 6S)-and 4-(2 ' S-4 '-methylthio group-2 '-(tert-butoxycarbonyl) amino butyryl radicals) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylic acid list sodium salt

Under the room temperature, 141mL (282mmol) 2N sodium hydroxide is added to (1R, 4S, 5S, 6S)-and 4-(2 ' S-4 '-methylthio group-2 '-(tert-butoxycarbonyl) amino butyryl radicals) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4 is in the solution that 6-diethyl dicarboxylate (58.95g, 112.8mmol theoretical value, preparation example 54) forms in the 141mL tetrahydrofuran (THF).Under the room temperature with mixture vigorous stirring 2 minutes.With 141mL t-butyl methyl ether diluting soln, separate each layer then.Further dilute water layer with 141mL water, and the dropping concentrated hydrochloric acid reduces to 4.46 with pH.Stir and obtained dilute slurry in 10 minutes.The denseer hydrochloric acid of adding is reduced to 1.4 with pH and (is used the dense HCl of 17mL altogether, 204mmol) in slurries.Stir after 2 hours, filter this slurries.(2 * 118mL) washing leaching cakes are delivered to the pan of a steelyard weighing before 45 ℃ of following vacuum-dryings 1 hour to water.Under 45 ℃ dry again 16 hours then, under 58 ℃ dry 5 hours again, obtain 52.96g (96% weight yield) title compound, be white solid.

258 ℃ of mp (decomposition).

[α] ²³D-25.2(c 1.03，H ₂O)。

500MHz ¹H NMR(D ₂O)δ4.07-4.01(m，2H)，3.45-3.43(m，1H)，3.11(d，1H，J＝15.0Hz)，2.85(m，1H)，2.71(s，3H)，2.47-2.35(m，3H)，1.96-1.90(m，4H)，1.78-1.72(m，1H)，1.28(s，9H)。

¹³C NMR(125 MHz，CD ₃OD)δ 174.45，173.58，172.80，157.46，81.71，61.41，55.04，53.24，42.29，31.71，30.85，29.41，27.79，23.65，14.41。FTIR(ATR)3287.62(s)，1698.00(s)，1528.91(s)，1327.36(s)，1283.74(s)，1245.90(s)，1174.81(s)，1109.06(s)，1053.05(s)，874.27(s)，808.95(s)cm ^-1。

Preparation example 56

(1R, 4S, 5S, 6S)-and 4-(2 ' S-4 '-methylthio group-2 '-(tert-butoxycarbonyl) amino butyryl radicals) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dicarboxylic acid

Under the room temperature, 397mL (795mmol) 2N sodium hydroxide is added to (1R, 4S, 5S, 6S)-and 4-(2 ' S-4 '-methylthio group-2 '-(tert-butoxycarbonyl) amino butyryl radicals) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4 is in the solution that 6-diethyl dicarboxylate (166.15g, 318mmol, preparation example 54) forms in the 480mL tetrahydrofuran (THF).With mixture vigorous stirring 2 minutes, reaction this moment became homogeneous phase and carries out under the room temperature, form clarifying faint yellow/green solution.Restir is 2 hours under the room temperature.With 480mL t-butyl methyl ether diluting soln, separate each layer then.Water layer is dropped to concentrated hydrochloric acid, and (71.5mL is 858mmol) in the solution that forms in water (960mL).Add ethyl acetate (500mL), add remaining water layer afterwards, obtain milk sap, use further dilution of ethyl acetate (460mL) again.Milk sap was stirred 40 minutes, and (2 * 250mL) wash for filtration and water.Separate each layer of filtrate and use ethyl acetate (500mL) reextraction water layer.With salt solution (75mL) washing blended organic layer, drying (MgSO ₄), filter and vacuum concentration, obtain 125.26g (84% calibration yield) title compound, be white foam shape thing.

Preparation example 57

2-bromo-2-fluoro-2-(3-oxocyclopentyl) ethyl acetate

Under-20 ℃, the 2.99g that 8.45mL (50.4mmol) chlorotriethyl silane is added in the anhydrous acetonitrile (100mL) activates in Zn (45.8mmol) suspension, and stirs 5 minutes.Add 8.0mL (57.3mmol) 2,2-two bromo-2-ethyl fluoroacetates, and under-20 ℃, stirred the mixture 90 minutes.Add 1.86mL (22.9mmol) 2-cyclopentenes-1-ketone, and the reaction mixture stirring is spent the night, make temperature slowly rise to room temperature.Add HCl 1N (125mL) and EtOAc (100mL).Use saturated NaHCO ₃(2 * 150mL), water (2 * 150mL) and salt solution (2 * 150mL) washing organic layers, use anhydrous MgSO ₄Drying is filtered and concentrating under reduced pressure.With EtOAc/ hexane (1: 8) is elutriant, with column chromatography purifying residuum, obtains title compound (5.36g, 88% total recovery), is colorless oil, is non-enantiomer mixture.

¹HNMR(300MHz，CDCl ₃)：1.34-1.41(m，6H)，1.91-1.95(m，1H)，2.04-2.68(m，11H)，3.12-3.31(m，2H)，4.33-4.43(m，4H)。

The activation of Zn: the dense HCl of 10mL is added in the suspension that 100g Zn powder forms in water (900mL).Under the room temperature mixture was stirred 20 minutes.The water outlet of inclining, and water (3 * 250mL), acetone (3 * 250mL) and ether (2 * 100mL) washing residuums.Under 35 ℃, the residuum drying under reduced pressure is spent the night.

Preparation example 58

(1RS, 5RS, 6RS)-6-fluoro-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester

Under 0 ℃, 13.0mL (75.0mmol) ethyl diisopropyl amine is added in 2.0g (7.5mmol) 2-bromo-2-fluoro-2-(3-oxocyclopentyl) ethyl acetate (preparation example 57) solution among the DMF (8mL), and at room temperature mixture is stirred and spend the night.Add HCl 1N (20mL), water (15mL) and EtOAc (75mL) solution.The extraction organic layer is used saturated NaHCO ₃(2 * 100mL), water (2 * 100mL) and salt solution (2 * 100mL) washing, use anhydrous MgSO ₄Drying is filtered and concentrating under reduced pressure.With EtOAc/ hexane (1: 4) is elutriant, and with column chromatography purifying residuum, obtain anti-: the title compound mixture (1.17g, 84% yield) along being 5: 1 is colorless oil.

¹HNMR(300MHz，CDCl ₃)：1.33(t，3H，J＝7.1Hz)，2.19-2.34(m，3H)，2.41-2.49(m，1H)，2.59(d，1H，J＝6.6Hz)，2.71-2.76(m，1H)，4.29(q，2H，J＝7.1Hz)。

Preparation example 59

(1RS, 2SR, 5RS, 6RS)-2-spiral shell-5 '-glycolylurea-6-fluoro-dicyclo [3.1.0] hexane-6-carboxylic acid

Under ice bath, will (1RS, 5RS, 6RS)-(0.55mL, mixture 0.55mmol) stirred 10 minutes in ethanol (1mL) 6-fluoro-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester (0.1g, 0.54mmol, preparation example 58) with 1N NaOH.Dripping 1N HCl in mixture is 1 until pH, and it is distributed between EtOAc and salt solution.With EtOAc aqueous phase extracted twice, and use MgSO ₄The dry organic layer that merges, concentrating under reduced pressure then.Under 60 ℃, with residuum, (NH ₄) ₂CO ₃(0.31g, 3.2mmol) and KCN (0.11g, 1.62mmol) mixture stirs in (2mL) at EtOH and water (1: 1) and spends the night.In ice bath, cool off, use 1N KHSO ₄Processing makes the mixture acidifying.Removal of solvent under reduced pressure, and residuum is dissolved among the MeOH, filter and vacuum concentration.Thick product can use without just being further purified.

Preparation example 60

(1R, 2S, 5R, 6R)-2-(2 ' R-tert-butoxycarbonyl amino-propionyl amino)-6-fluoro-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate and (1S, 2R, 5S, 6S)-2-(2 ' R-tert-butoxycarbonyl ammonia Base-propionyl amino)-and 6-fluoro-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

Isomer A isomer B

With 1.59g (5.0mmol) Ba (OH) ₂Add to 0.32g (1.5mmol) (1R ^*, 2S ^*, 5R ^*, 6R ^*In the solution that)-2-spiral shell-5 '-glycolylurea-6-fluoro-dicyclo [3.1.0] hexane-6-carboxylic acid (preparation example 59) and water (10mL) form, and under 105 ℃, the mixture stirring is spent the night.Under 0 ℃, with 1NHCl solution being acidified to pH is 1, then concentrating under reduced pressure.With absolute anhydrous EtOH dissolving residuum and concentrating under reduced pressure several times until the solid complete drying.Under 0 ℃, with 0.37mL (5.0mmol) SOCl ₂Add in the residuum that is dissolved among the anhydrous EtOH, reflux then and stirred 5 hours.Use saturated NaHCO ₃Alkalizing solution, and add EtOAc (25mL).Separate organic layer, with more EtOAc (2 * 25mL) aqueous layer extracted.Use anhydrous MgSO ₄The organic layer of dry mixed filters and concentrating under reduced pressure.Residuum is dissolved in the 20mL mixture of DCM-DMF (4: 1), 0.73g (1.9mmol) HATU, 0.26g (1.9mmol) HoAt, 0.35g (1.8mmol) L-Ala.Adding 2.7mL (15.3mmol) diisopropyl ethyl amine also spends the night the mixture stirring under Ar atmosphere.Add DCM (15mL), separation is also used saturated NaHCO ₃(2 * 25mL), water (2 * 25mL) and salt solution (2 * 25mL) washing.Use anhydrous MgSO ₄Dry organic layer, filtration and concentrating under reduced pressure.With EtOAc/ hexane (1: 2) is elutriant, with column chromatography purifying residuum, obtains title compound (0.33g, 51% total recovery), and it is 1: 1 a non-enantiomer mixture, is colorless oil.

Use chirality HPLC, separate non-enantiomer mixture according to following analytical procedure: ChiralpakAD 10m, 4.6 * 250mm; Elutriant: the 10%IPA in the hexane; Flow velocity: 1.0mL/ minute; UV:215nm.Isomer A retention time=5.9 minutes.Isomer B retention time=9.2 minutes.

Isomer A:(1R, 2S, 5R, 6R, 2 ' R)-2-(2 '-tert-butoxycarbonyl amino-propionyl amino)-6-fluoro-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

¹HNMR(300 MHz，CDCl ₃)：1.18(t，3H，J＝7.1Hz)，1.23(t，1H，J＝7.1Hz)，1.36(s，9H)，1.56-1.68(m，1H)，2.03-2.11(m，2H)，2.21-2.31(m，1H)，2.37-2.45(m，1H)，2.61-2.63(m，1H)，4.09-4.16(m，5H)，5.10(bd，1H，J＝6.6 Hz)，7.11(bs，1H)。

¹³C-NMR (75MHz, CDCl ₃): 13.9,14.0,17.7,24.9 (d, J=1.0Hz), 28.1,31.5 (d, J=10.9Hz), 34.9 (d, J=9.4Hz), 36.4 (d, J=8.8Hz), 49.1,61.5,61.8,67.4,77.2,81.5 (d, J=242.6Hz), 155.4,168.6 (d, J=25.1Hz), 171.8 and 172.7ppm.

Isomer B:(1S, 2R, 5S, 6S, 2 ' R)-2-(2 '-tert-butoxycarbonyl amino-propionyl amino)-6-fluoro-dicyclo [3.1.0] hexane-2, the 6-diethyl dicarboxylate

¹HNMR(300 MHz，CDCl ₃)：1.13(t，3H，J＝7.1Hz)，1.18(t，1H，J＝7。1Hz)，1.31(s，9H)，1.51-1.60(m，1H)，1.98-2.07(m，2H)，2.15-2.22(m，1H)，2.26-2.39(m，1H)，2.55-2.60(m，1H)，4.07-4.12(m，5H)，5.20(sd，1H)，7.22(bs，1H)。

¹³C-NMR (75 MHz, CDCl ₃): 13.7,13.8,18.0,24.7,28.0,31.4 (d, J=10.9Hz), 34.5 (d, J=8.9 Hz), 36.3 (d, J=9.4Hz), 49.1,61.4,61.7,67.3,77.2,81.4 (d, J=242.6Hz), 155.2,168.5 (d, J=25.1 Hz), 171.7 and 172.8ppm.

Preparation example 61

(1R, 2S, 5R, 6R)-and 2-amino-6-fluoro-dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

Placing 6N HCl (2mL) to reflux the isomer A in 30mg (0.07mmol) preparation example 60 spends the night.Removal of solvent under reduced pressure with the ether washing, is dissolved among the MeOH (1mL), and adds propylene oxide (propilene oxide) (2mL).Stir under the room temperature and spend the night.Incline and desolvate, wash residuum, and use the Ar air stream drying, obtain title compound (12mg, 85%), be white solid with ether.

[α]D＝-25.0(c＝0.80mg/mL，H ₂O)。

Preparation example 62

(1S, 2R, 5S, 6S)-and 2-amino-6-fluoro-dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

Isomer B with preparation example 60 is a starting raw material, and the method according to preparation example 61 prepares title compound basically.

[α]D＝21.7(c＝0.46mg/mL，H ₂O)。

Preparation example 63

(6S)-4-thia dicyclo [3.1.0] oneself-2-alkene-6-carboxylic acid, ethyl ester

On 5 liters three-necked bottle, agitator is installed, thermopair, a tetrafluoroethylene adds alkene pipe and a N ₂Inlet, 2000mL thiophene (d=1.05g/ml packs into, begin to be 25.0mol, along with the solution of ethyl diazoacetate in thiophene adds and increases to 45mol, based on the total amount of the ethyl diazoacetate that adds and not proofread and correct EDA usefulness be 17.1 equivalents, be 19.0 equivalents) based on gauged ethyl diazoacetate.Under the nitrogen atmosphere, to wherein adding the sad rhodium [Rh of 0.968g ₂(oct-anoate) ₄] (1.24mmoles is 0.0472mol% based on the ethyl diazoacetate of school usefulness not, is 0.052mol% based on the mole number of pure ethyl diazoacetate, Johnson Mathey:lot.No.059255001).Suspension is heated to 46 ℃, and stirred 10 minutes down, thereby be dissolved as green solution at 46 ℃.Through positive-displacement pump, and adding 300g ethyl diazoacetate in this solution (90% is pure, and 2.63 moles, 2.37 moles of pure product, 1.00 equivalents, Aldrich: lot number (LotNo.) 17603PI) be dissolved in formed solution in the 1600mL thiophene.It is 8 hours that adding speed should make total joining day; Adding speed suppresses the formation of ethyl maleate and ethyl fumarate slowly.Do not have ethyl diazoacetate when residue (about 30 minutes), with dark amber reactant (3,985g) be cooled to 23 ℃.Reaction mixture is divided into several parts, and more part (3,240g/3,985g=81.3%) direct simmer down to oily matter.1.5 holder, under 23 ℃, make crude product, make the product degassing and remove remaining thiophene by changeing distillation device.Then 120 ℃ and 1.3 the holder under product of distillation.Target product (faint yellow) is collected as two batches of cuts, 155.0g and 32.2g.Analyzing definite two batches of usefulness separately through HPLC is 78% and 76%.Above-mentioned overhead product is carried out crystallization: it is dissolved in methyl alcohol (2mL/g overhead product) and is cooled to-10 ℃, and can not choose and to solution in add crystal seed if observe crystal growth this moment.In case crystal begins growth, just mixture further is cooled to-45 ℃ and stirred 2-3 hour, filtration is also washed with cold (45 ℃) methyl alcohol (1 * 1mL/g overhead product).24 ℃ of these materials of following vacuum-drying obtain title compound, are white-near-white solid, rate of recovery 80-85%, usefulness＞98%.

Preparation example 64

(4S, 6S)-4-hydroxyl-2-thia dicyclo [3.1.0] hexane-6-hydroxy acid ethyl ester

In 15-20 minute, under 0 ℃, nitrogen atmosphere, (6S)-4-thia dicyclo [3.1.0] oneself-2-alkene-6-carboxylic acid, ethyl ester (22.2g, 131mmol) add in the solution that in the 136mL tetrahydrofuran (THF), forms borine-THF complex compound (98mL, 98mmol).0 ℃ is stirred after 30 minutes down, makes reaction be warming up to 15 ℃ and stir until HPLC and judge react completely (1.5-2 hour).Reaction is cooled to 0 ℃, and is that keeping temperature is 0 ℃ in 7 the buffered soln with 10-15 branch clock time its 1N pH that is transferred to 111mL pre-cooled (0 ℃).Branch adds the Sodium peroxoborate monohydrate for five times in mixture (15.6g, 157mmol) solid is so that temperature remains below 20 ℃.Make solution be warming up to room temperature and stirred 1 hour, add 222mL water subsequently.Stir after 2 hours, add Sulfothiorine pentahydrate (9.7g) the cancellation superoxide that is dissolved in the 24mL water, stirred afterwards 10 minutes.With ethyl acetate (2 * 222mL) extraction mixtures.(1 * 222mL) washing blended organic extract, (1 * 222mL) washs, and vacuum concentration is to dry to use salt solution afterwards with saturated sodium bicarbonate aqueous solution.Thick product is dissolved in 1, in the 2-ethylene dichloride (1mL/1g is oil slightly), and goes up silicagel column (4g silica gel/1g pulping and the thick oil that is seated in 15% ethyl acetate-heptane).With 15% ethyl acetate-heptane wash-out pillar, until product on TLC as seen, this moment solvent is become 50% ethyl acetate-heptane.All are contained the partially mixed of target product, and vacuum concentration is an oily matter.Total recovery is 55-65%.

Preparation example 65

(6S)-4-oxo-2-thia dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester

In 30 minutes, under-70 ℃, methyl-sulphoxide (33.4mL, 471mmol) slowly add trifluoroacetic anhydride (33.2mL, 235mmol) solution that in the 73mL methylene dichloride, forms (temperature remains below-66 ℃) in the solution that in the 194mL methylene dichloride, forms.Stir after 20 minutes, with 60 fens clock times to wherein add (4S, 6S)-(34.1g, the 181mmol) solution that forms in the 194mL methylene dichloride is so that temperature remains below-60 ℃ to 4-hydroxyl-2-thia dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester.Stir after 1 hour, (75.7mL 543mmol) handles reactant 35 minutes, so that temperature remains below-50 ℃ with triethylamine.Restir reaction 1 hour is removed cooling bath and add the hydrochloric acid of 400mL2N this moment.After being warming up to 0 ℃, separate each layer, and with 2N hydrochloric acid (1 * 300mL), the 1N sodium bicarbonate aqueous solution (1 * 670mL) and water (1 * 300mL) washing organic layer is used dried over sodium sulfate afterwards, filter, and vacuum concentration is red oil that it leaves standstill to produce and solidifies.Allow unmodified packed column (2g/1g has filled the initial alcohol of methylene dichloride) on the crude product, and with methylene dichloride (200-300mL) wash-out.Collect all parts that contain product and concentrated, obtain target product, be the orange/brown solid.Usually yield corrected is 85-90%.

Preparation example 66

(6S, 11S)-8,10-dioxo-2-thia spiral shell [dicyclo [3.1.0] hexane-4,5 '-imidazolidine]-6-carboxylic Acid

With volatile salt (2.46g, 25.6mmol) and potassium cyanide (0.817mg 12.5mmol) is blended in the 19.9mL methyl alcohol, stirs 30 minutes.(2.39g, 12.8mmol) solution-treated mixture is heated to 30 ℃ with reaction, and stirs 23 hours with (the 6S)-4-oxo in the 19.9mL methyl alcohol-2-thia dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester.Evaporate volatile matter, residuum is dissolved in the 2.75N sodium hydroxide (13.1mL), stirred 1 hour.After the dilution of 13.1mL water, with concentrated hydrochloric acid pH is reduced to 3.1, and optional so that (6S, 11R)-8, [dicyclo [3.1.0] hexane-4,5 '-imidazolidine]-the 6-carboxylic acid is a crystal seed to 10-dioxo-2-thia spiral shell.PH is reduced to 1.0, suspension is cooled to 0 ℃, and stirred 1.25 hours.Collect brown solid,, and spend the night 40 ℃ of following vacuum-dryings with cold water (2.3mL and 0.8mL) washing, obtain 2.00g (55% corrected purity) (6S, 11R)-8,10-dioxo-2-thia spiral shell [dicyclo [3.1.0] hexane-4,5 '-imidazolidine]-6-carboxylic acid.Handle with 50mL ethyl acetate dilution filtrate and with 18mL sodium-chlor.Stir after 15 minutes, separate each layer, further use ethyl acetate (2 * 50mL) washing water layers.With dried over sodium sulfate blended organic layer, filter, vacuum concentration is slurries (about 5mL), to wherein adding t-butyl methyl ether (25mL), stirs afterwards and spends the night.Collect solid, with the t-butyl methyl ether washing, and 40 ℃ of following vacuum-dryings 2 hours, obtain 0.64g (10% corrected purity) title compound, it is 1: 1 mixture of non-mapping glycolylurea.Second batch of glycolylurea and first glycolylurea are mixed, carry out following step.

Split

In the solution that racemic tartaric acid (15g, 65.7mmol, about 6: 1 of the ratio of first urea in the non-mapping second) forms in 300mL ethanol and 75mL water, add (R)-phenyl glycinol (9.0g, 65.7mmol) in.With mixture heating up to about 80 ℃ to promote dissolving.This dark solution is slowly cooled off, and 40-45 ℃ can be observed precipitation.These slurries further are cooled to 0 ℃ and kept 1-1.5 hour.Collect solid, with 4: 1 ethanol: water (1 * 60mL is precooled to 0 ℃) washing (stirring simultaneously), 65 ℃ following vacuum-drying 12-24 hour.Usually the productive rate of the salt that splits is 37-45%, and observe＞98%de and＞98%ee.The salt that splits is dissolved in 6 volumes (mL/g) water, handles with 1.1 normal dense HCl afterwards.Slurries are cooled to 0 ℃ and stirred 1 hour, filter afterwards, with the cold rinse of 1 volume, and 60 ℃ of following vacuum-dryings.Common title compound productive rate＞90%, and %de and %ee＞99%.

Preparation example 67

(6S, 11S)-2,2,8,10-four oxos-2-thia spiral shell [dicyclo [3.1.0] hexane-4,5 '-imidazolidine]-6- Carboxylic acid

In 6.8mL water, 0.7mL50% aqueous sodium hydroxide solution and 186mg (0.74mmol) wolframic acid, add (6S, 11S)-8,10-dioxo-2-thia spiral shell [dicyclo [3.1.0] hexane-4,5 '-imidazolidine]-6-carboxylic acid (3.4g, 14.9mmol).Gained solution is heated to 50 ℃, and in 66 minutes, (7.7mL handles slowly 74.5mmol) with 35% hydrogen peroxide.47-48 ℃ of following stirring reaction 5 hours, be cooled to 0 ℃ subsequently afterwards, use the skim diatomite filtration, and with cold water (1 * 2mL) rinsing.Filtrate is heated to 50 ℃, handles making pH=1.5 with concentrated hydrochloric acid.Making slurries be cooled to room temperature and stir spends the night.After being cooled to 0 ℃, filter slurries, usefulness cold water (2 * 2mL) washings, 55 ℃ of following vacuum-dryings get 3.19g (82%) title compound to constant weight, are white solid:

[α] ²⁵ _D-48.6(c 1.19，1N NaOH)。

275 ℃ of mp (grey), 295 ℃ (brown).

500MHz ¹H NMR(DMSO-d ₆)δ13.15(br s，1H)，10.99(s，1H)，8.13(s，1H)，3.85(d，1H，J＝15.0Hz)，3.74(dd，1H，J＝7.0，4.0Hz)，3.03(d，1H，J＝15.5Hz)，2.80(dd，1H，7.0，4.0Hz)，2.39(t，1H，J＝4.0Hz)。 ¹³C NMR(125MHz，DMSO-d ₆)δ174.39，169.87，156.35，62.67，52.59，44.16，31.69，21.92；FTIR(KBr)3317(s)，3250(s)，3211(s)，3086(w)，1791(s)，1742(s)，1713(s)，1327(s)，1192(s)，1140(s)cm ^-1。

C ₈H ₈N ₂O ₆The analytical calculation value of S: C, 36.93; H, 3.10; N, 10.77.Measured value: C, 36.76; H, 3.07; N, 10.60.

Preparation example 68

(1R, 4S, 5S, 6S)-4-[(2 ' S)-(2 ' amino)-propionyl] amino-(the 2-sulphonyl dicyclo [3.1.0] of mixing Hexane)-4,6-dicarboxylic acid

In stainless steel Parr reactor, add (6S, 11S)-2,2,8,10-four oxos-2-thia spiral shell [dicyclo [3.1.0] hexane-4,5 '-imidazolidine]-6-carboxylic acid (2.50g, 9.60mmol) and 2N sodium hydroxide (24.0mL, 48.0mmol).Mixture heating up to 95 ℃ and stir 21 hours after, again mixture is cooled to room temperature and handles with activated carbon (1.25g).Use the diatomite filtration mixture, and, use H again distillate simmer down to 17g ₂O dilute weight 24g.With dense HCl pH is reduced to 6.5, and with mixture heating up to 62 ℃.After with dense HCl pH being reduced to 2.5, crystallization takes place.Make suspension be cooled to 30 ℃ earlier,, cool the temperature to 5 ℃ more again with pH regulator to 1.7.Suspension is collected solid and is used cold H after keeping 18 hours under this temperature ₂O (2 * 2.9mL) washings.45 ℃ of these white solids of following vacuum-drying obtain title compound (1.81g, 80%).Make title compound pulping and at 3-4 hour internal heating to 85 ℃ in 10 volume water, be cooled to room temperature, stirred 2-3 hour, filter, and water (1 * 1 volume) washing.The rate of recovery＞95%.

Preparation example 69

(6S)-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester

Under the room temperature, (134g 585mmol) dripped 87.4mL (585mmol) 1 in the suspension that forms in the 486mL acetonitrile, 8-diazabicyclo [5.4.0] 11 carbon-7-alkene to bromination (ethoxy carbonyl methyl) dimethyl sulfonium with 15 fens clock times.Stir after 1 hour, handled yellow mixture 10 minutes with 40g (487mmol) 2-cyclopentenes-1-ketone.The mixture stirring is spent the night, add 480mL tertiary butyl ethyl ether again, (1 * 240mL) washing, (1 * 240ml) washes water layer with methyl tertiary butyl ether to use 1N hydrochloric acid subsequently.With salt solution (organic extract, drying (MgSO that 1 * 400mL) washing merges ₄), filter, and vacuum concentration obtains thick (6S)-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester, is orange solids (84.8g).Can be by distillation (～138 ℃ make solidified overhead product pulping, filtration, drying in heptane come the thick material of purifying 10mmHg), subsequently.

Preparation example 70

(±) (6S)-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid

Under the room temperature, in the solution that thick (6S)-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester (30.2g, 180mmol do not proofread and correct) forms, add 89mL (178mmol) 2N sodium hydroxide in 30mL ethanol.Stir after 80 minutes, with t-butyl methyl ether (1 * 90mL) washing reaction mixture, and make pH=1.0 with concentrated hydrochloric acid (18mL) processing water layer.With 15g sodium-chlor treating mixture, use ethyl acetate (3 * 90mL) washings afterwards.Organic extract (the Na of dry mixed ₂SO ₄), filtering, vacuum concentration obtains 23.8g (94%, do not proofread and correct) title compound, is the near-white solid.

Preparation example 71

(±) (6S)-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid N-benzyl-alpha-methyl-benzyl amine salt

Under reflux temperature, thick (±) (6S)-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid (11.9g, 84.9mmol, suppose 100% usefulness) at 119mL6: 1 ethyl acetate: add in the solution that forms in the ethanol 18g (85.1mmol) (S)-N-benzyl α-Jia Jibianji amine.After the dissolving, mixture is cooled to 52 ℃ and optional add crystal seed subsequently under this temperature.Be cooled to room temperature, and behind the restir 13.5h, collect crystal and with 6: 1 ethyl acetate: (2 * 48mL) wash ethanol.Vacuum-drying, obtain 10.8g (36%, the 77%de) salt of Chai Fening, it is a solid.

Analyze to determine the de of salt by the chirality GC of deutero-methyl ester, it is prepared as follows: the salt that 150mg is split is dissolved in the 5mL methylene dichloride, with (2 * 1mL) washings of 1N sulfuric acid.Dry organic layer filters, and dilutes with 2mL methyl alcohol, and handles with the 1mL 2M trimethyl silyl diazomethane in the hexane.Stir under the room temperature after 15 minutes, vacuum concentrated mixture obtains being suitable for the methyl ester that chirality GC analyzes.GC condition: 30m * 0.25mm * 0.25 μ β-DEX, 325 posts, 140 ℃ of oven temperatures, Hai Zaiqi @1mL/ minute, 250 ℃ of following FID detected, 1 μ L part 1: 100, Shi Yang @1mg/mL is in methylene dichloride.

Preparation example 72

(6S)-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester

To 46.3g (132mmol) (6S)-add in 2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid N-solution that benzyl-alpha-the methyl-benzyl amine salt forms in the 200mL ethyl acetate

198mL (198mmol) 2N sodium hydroxide.Behind the good mixing, separate and be equipped with layer, and (1 * 200mL) washs water layer with ethyl acetate.Handle water layer with 18mL (211mmol) concentrated hydrochloric acid and 100g sodium-chlor.Stirred the mixture 30 minutes, and used ethyl acetate (2 * 200mL) washings afterwards.Dry organism (the MgSO that merges ₄), filtering, vacuum concentration obtains the acid that 18.3g (99%) splits [(+) (6S)-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid], is white solid.

Next, the thick fractionation acid product that 10g (71mmol) is above-mentioned is dissolved in the 42mL ethanol, and dropping 4mL (71mmol) vitriol oil is handled.With mixture heating up to 45 ℃ and stirred 75 minutes.After being cooled to room temperature, adding 45mL water, and add 20mL ethyl acetate and 12g sodium bicarbonate together.After stirring several minutes, with ethyl acetate (2 * 50mL) purging compounds.Dry organism (the MgSO that merges ₄), filtering, vacuum concentration gets thick (6S)-2-oxo dicyclo [3.1.0] hexane of 11g (92%)-6-carboxylic acid, ethyl ester, is white solid.By 6: 1/ heptane: crystallization got title compound in the t-butyl methyl ether (3.5mL/g substrate), and productive rate is about 80%, and by chirality GC analysis＞98%ee.

Preparation example 73

(6S)-6-(ethoxy carbonyl) dicyclo [3.1.0] oneself-2-alkene-2-yl acetate

Use the Dean-Stark device, with (6S)-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester (380.1g in the pseudoallyl acetic ester (2.26L), 2.26mol) and sulfuric acid (18M, 6.3mL, 0.11mol) mixture heating up refluxed 2.5 hours, this moment, GC analyze to show, title compound with (6S)-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester is 9: 1 a mixture.Distill removed the 950mL solvent in 1 hour after, GC shows that product/initial substance ratio is 17: 1.Add other pseudoallyl acetic ester (900mL) and dense H ₂SO ₄(3.15mL), the restir mixture is 15 hours under reflux temperature, and this moment, GC showed 27: 1 product/initial substances.After other 1.35L solvent is removed in distillation, mixture is cooled to room temperature, uses MTBE (2L), H then ₂O (250mL) and saturated NaHCO ₃The aqueous solution (800ml) dilution.Separate each layer, with salt solution (400mL) washing organic layer.With the water layer that MTBE (400mL) extraction merges, the dry organic layer (Na that merges ₂SO ₄), filter and be concentrated into dark red/brown oil (540g).With thick oil content is that two five equilibriums are also used quick SiO ₂Layer (every crowd of 713g) filters, with 10: 1/ heptane: eluent ethyl acetate.To partly combine and concentrate from the product that closes of two packed columns, obtain target mixture, be yellow oil (460g, 97%; After with NMR solvent being proofreaied and correct is 90%).Make the analytical pure sample of title compound with the silica gel column chromatography of ethyl acetate/hexane (1: 5) wash-out, be colorless oil.

[α] ²⁵ _D+185(c 1.48，CHCl ₃)。

500MHz ¹H NMR(CDCl ₃)δ5.19-5.18(m，1H)，4.12(q，1H，J＝7.0Hz)，4.11(q，1H，J＝7.0 Hz)，2.74-2.69(m，1H)，2.48-2.43(m，2H)，2.22-2.19(m，1H)，2.16(s，3H)，1.39(dd，1H，J＝2.5，2.5Hz)，1.25(t，3H，J＝7.0Hz)。

¹³C NMR(125 MHz，CDCl ₃)δ 173.37，169.01，152.26，111.56，61.28，32.47，32.40，29.72，24.97，21.67，14.95。

FTIR(CHCl ₃)3026(m)，2985(m)，1724(s)，1272(s)，1187(s)cm ^-1。

C ₁₁H ₁₈NO ₄[M+NH ₄] ⁺ES HRMS calculated value 228.1236, measured value 228.1252.

Preparation example 74

(3S, 1R, 6R)-7-oxa--5-oxo three rings [4.1.0.0＜2,4 〉] heptane-3-carboxylic acid, ethyl ester

With 2.02L 1, (6S)-6-(ethoxy carbonyl) dicyclo [3.1.0] in the 4-dioxane oneself-2-alkene-2-yl acetate (212.2g, 1.01mol) and 2,3-two chloro-5,6-dicyano-1, and the 4-benzoquinones (252.0g, 1.11mol) mixture heating up is to refluxing, and stirred 17 hours, this moment GC analyze to show be converted into fully (6S)-4-oxo dicyclo [3.1.0] oneself-2-alkene-6-carboxylic acid, ethyl ester.Mixture is cooled to room temperature and uses THF (564mL) dilution.After mixture is cooled to 8 ℃, add 1 with 30 fens clock times, and 8-diazabicyclo [5.4.0] 11 carbon-7-alkene (377mL, 2.52mol), so that temperature of reaction remains below 10 ℃.Mixture is cooled to 5 ℃ then, and (70wt%, in the water, 210mL 1.51mol), keeps temperature of reaction to be lower than 9 ℃ to add tertbutyl peroxide in 50 minutes.After 50 minutes, also (2 * 800mL) wash brown filter cake to the filtering reaction thing with MTBE with the mixture restir.In filtrate, add 1.20L 1NHCl, behind the good mixing, separate each layer.Use saturated NaHCO successively ₃(1.20L) aqueous solution, saturated Na ₂S ₂O ₃(1.20L) aqueous solution and salt solution (600mL) washing organic layer.Solution drying (Na ₂SO ₄) after, the orange mud shape of simmer down to settling, and dilute with the 200mL heptane.Evaporate volatile matter, make orange solids, grind, filter, with other heptane (2 * 175mL) washing leaching cakes with the 350mL heptane.With the solid vacuum-drying that is collected 17 hours, make 138.7g (75%) title compound under the room temperature, be pale brown look solid.Make the analytical pure sample of title compound by crystallization among the MTBE, be white solid.

[α] ²⁵ _D+ 2.3 (c 1.20, CHCl ₃) ,+8.4 ° (c 1.28, acetone); Mp 129-130 ℃.

500MHz ¹H NMR(CDCl ₃)δ 4.16(q，2H，J＝7.0Hz)，3.99(t，1H，J＝2.5Hz)，3.24-3.23(m，1H)，2.96-2.94(m，1H)，2.21-2.19，(m，1H)，2.08(t，1H，J＝3.0Hz)，1.26(t，3H，J＝7.0Hz)。

¹³C NMR(125 MHz，CDCl ₃)δ201.19，168.84，62.42，57.04，51.25，31.16，30.54，29.60，14.79。

FTIR(KBr)3087(w)，3059(w)，3051(w)，3007(w)，2993(w)，2963(w)，1753(s)，1719(s)，1273(s)，1191(s)，1009(m)，848(m)cm ^-1。

C ₉H ₁₀O ₄The analytical calculation value: C, 59.34; H, 5.53.Measured value: C, 59.32; H, 5.43.

Preparation example 75

(6S)-4-oxo dicyclo [3.1.0] oneself-2-alkene-6-carboxylic acid, ethyl ester

Though the common original position of title compound is used for preparation (3S, 1R, 6R)-7-oxa--5-oxo three ring [4.1.0.0＜2,4 〉] heptane-3-carboxylic acid, ethyl ester, but also can obtain the analytical pure sample of title compound: filter the reaction mixture that contains this compound, and the evaporation filtrate is to obtain brown solid.This solid is suspended in the ethyl acetate again filtering suspension liquid and concentrating filtrate.Residuum through silica gel and ethyl acetate/hexane (1: 5-1: 2) chromatography makes title compound, and it is come out by recrystallization in the hot ethyl acetate, again by the condition of front through chromatography, obtain title compound, be white solid.

[α] ²⁵ _D-268(c 1.17，CHCl ₃)。

mp 97-98℃。

500MHz ¹H NMR(CDCl ₃)δ 7.60(ddd，1H，J＝5.5，2.5，0.75Hz)，5.73(dd，1H，J＝5.0，0.5Hz)，4.15(q，2H，J＝7.0Hz)，2.96-2.94(m，1H)，2.63-2.61(m，1H)，2.60(t，1H，J＝2.5Hz)，1.26(t，3H，J＝7.0Hz)。

¹³C NMR(125 MHz，CDCl ₃)δ 203.96，168.61，160.33，130.29，62.03，46.53，30.72，29.62，14.82。

FTIR(KBr)3080(m)，2996(m)，1717(s)，1695(s)，1266(s)，1291(m)，1191(s)，1179(s)cm ^-1。

C ₉H ₁₀O ₃The analytical calculation value: C, 65.05; H, 6.07.Measured value: C, 64.97; H, 6.01.

Preparation example 76

(4S, 6S)-4-hydroxyl-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester

Use sodium-acetate (36.1g successively, 0.44mol), sodium iodide (65.8g, 0.44mol) and acetate (27.5mL, 0.48mol) handle (3S under the agitation condition, in the 667mL acetone, 1R, 6R)-7-oxa--5-oxo three rings [4.1.0.0＜2,4 〉] and heptane-3-carboxylic acid, ethyl ester (36.3g, 0.20mol).Stirred the mixture under 30 ℃ 15 hours, vacuum is removed acetone afterwards, and the residue brown solid makes solid at ethyl acetate (323mL) and H ₂Distribute between the O (323mL).Separate each layer, with ethyl acetate (3 * 323mL) washing water layers.Use saturated Na successively ₂S ₂O ₃(364mL) aqueous solution and saturated NaHCO ₃(364mL) organism of solution washing merging.With the each wash water solution of ethyl acetate (323mL) reextraction.Dry organism (the Na that merges ₂SO ₄), filter and simmer down to red-brown oil, it is dissolved in the 300mL ethanol.The evaporation volatile matter obtains target product, and it is red-brown oil (41.8g, 114%).Through ethyl acetate/hexane (1: 2～2: 1) silica gel column chromatography, afterwards again by recrystallization among the MTBE, obtain the analytical pure sample of title compound, be white solid.

[α] ²⁵ _D+3.9(c 1.39，CHCl ₃)，+6.0°(c 1.69，MeOH)。

mp 81-82℃。

500MHz ¹HNMR(CDCl ₃)δ 4.60(br s，1H)4.1 6(q，2H，J＝7.0Hz)，2.66(dd，1H，J＝5.0，4.0Hz)，2.42-2.40(m，1H)，2.34(dd，1H，J＝19.0，5.5Hz)，2.24，(br d，1H，J＝3.0Hz)，2.07(d，1H，J＝19.0Hz)，1.91(t，1H，J＝3.0Hz)，1.27(t，3H，J＝7.0Hz)。

¹³C NMR(125MHz，CDCl ₃)δ209.74，170.07，69.04，62.32，43.47，36.89，34.95，26.14，14.83。

FTIR(CHCl ₃)3607(w)，3447(w)，3025(m)，2985(w)，1739(s)，1728(s)，1270(s)，1187(s)cm ^-1。

C ₉H ₁₂O ₄The analytical calculation value: C, 58.69; H, 6.57.Calculated value: C, 58.48; H, 6.63.

Preparation example 77

2-[((1R)-1 phenylethyl) amino] (2S, 4S, 6R)-2-cyano group-4-hydroxyl dicyclo [3.1.0] oneself Alkane-6-carboxylic acid, ethyl ester

To ethanol (332mL) and H ₂(4S among the O (332mL), 6S)-4-hydroxyl-2-oxo dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester (68.2g, because ethanol impurity, calibrated is 60.0g, 0.326mol) add (R)-methyl-benzyl amine (46.3mL in the solution, 0.359mol) and NaCN (20.8g 0.424mol), keeps temperature between 20-25 ℃.Then with 10 fens clock times add dense HCl (35.3mL, 0.424mol), during keep above-mentioned temperature of reaction.Dark brown mixture was stirred 1 hour, and optional afterwards is that crystal seed causes crystallization with the title compound.Suspension was stirred 1 hour, add H afterwards ₂O (664mL).With suspension restir 1.75 hours, collect title compound, it is a brown solid, uses H ₂O (332mL) washing.Make the wet cake of air from strainer and passed through 25 minutes, then this material is directly used in nitrile hydrolysis (the heavy 145g of wet cake).Though the temperature in the process of vacuum drying is higher than 25 ℃, then title compound decomposes rapidly, vacuum-drying small sample at room temperature and do not produce decomposition.

[α] ²⁵ _D+81.6(c 1.18，CHCl ₃)。

mp 70-72℃。

500MHz ¹H NMR(CDCl ₃)δ 7.39(d，2H，J＝7.0Hz)，7.26-7.16(m，3H)，4.31(d，1H，J＝5.0Hz)，4.22(q，1 H，J＝6.5Hz)，3.93-3.85(m，2H)，2.33(d，1H，J＝15.0Hz)，2.01(br t，1H，J＝4.5Hz)，1.64(dd，1H，J＝15.0，5.0Hz)，1.55-1.54(m，1H)，1.40-1.39(m，4H)，1.17(t，3H，J＝7.0Hz)。

¹³C NMR(125MHz，CDCl ₃)δ170.54，144.85，128.61，127.45，127.38，121.88，72.17，61.02，60.66，56.57，45.82，36.70，34.45，25.83，21.75，14.22。

FTIR(KBr)3568(m)，3489(m)，3285(m)，2923(m)，2228(w)，1712(s)，1298(m)，I 197(m)cm ^-1。

C ₁₈H ₂₃N ₂O ₃[M+H] ⁺FAB HRMS calculated value 315.1709, measured value 315.1704.

Preparation example 78

2-[((1R)-and the 1-phenylethyl) amino] (2S, 4S, 6R)-and 4-hydroxyl dicyclo [3.1.0] hexane-2,6-two Carboxylic acid

2-[((1R in DMSO (220mL))-and the 1-phenylethyl) amino] (2S, 4S, 6R)-slowly add 30%H in 2-cyano group-4-hydroxyl dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester wet cake (0.326mmol theoretical value) solution ₂O ₂(44.5mL 0.426mol), keeps temperature to be lower than 27 ℃.Cool the temperature to 19 ℃, (52.3mL, 0.262mol), keeping temperature is 22-27 ℃ at first carefully to add 5N NaOH lentamente with 15 fens clock times.Need to use suitable volumetrical ice bath processing reaction heat.Stirring in the said temperature scope should be brown, homogeneous mixture is after 20 minutes, and HPLC demonstration initial substance is consumed, has generated amide intermediate.The reaction mixture restir after 1.5 hours, is added Na ₂SO ₃(13.7g 0.109mol), and stirred the mixture 15 minutes, tested that it is negative to superoxide this moment with starch-iodide test paper.Add 3N NaOH (291mL, 0.873mol) after, with mixture heating up to 85 ℃ and stirred 18 hours.The homogeneous brown mixture is cooled to 30 ℃, adds dense HCl and make pH reduce to 3.6, keeping temperature is 30-35 ℃.PH is that crystallization in 3.6 o'clock begins, and restir suspension is 15 minutes afterwards, then pH is reduced to 2.5.With the mixture restir after 10 minutes, be cooled to 2 ℃ and stirred 2 hours, collect gray solid afterwards and use cold H ₂O (400mL) and EtOH (300mL) washing.With the solid vacuum-drying of collecting 17 hours, obtain 42.9g (43%, calculate) title compound under 45 ℃ by the initial substance of preparation example 18.In order further to process the title compound that makes in the reaction, in the following manner by reclaiming in the mother liquor.Ethanol part in the mother liquid evaporation combines residuum with aqueous solution part in the mother liquor.Remove H under reduced pressure ₂O (485mL) uses 70mL5N NaOH and 5mL50%NaOH with mother liquor pH regulator to 12.9.With n-BuOH (3 * 800mL) washing solns, with dense HCl with pH regulator to 2.5, and concentrated solution.Dilute residuum and evaporate (2 *) volatile matter with EtOH (100mL).Dilute residuum with EtOH (150mL), and wash the brown solid that contains other title compound and salt with EtOH (75mL), 50 ℃ of following vacuum-drying to weight are 102g.The two batches of title compounds all are used in subsequently the esterification.

[α] ²⁵ _D+4.5(c 1.41，1N NaOH)。

220 ℃ of mp (becoming grey) by near-white, 280 ℃ (brown).

500MHz ¹H NMR(D ₂O，KOD)δ7.39(d，2H，J＝7.0Hz)，7.19-7.04(m，5H)，3.92(d，1H，J＝5.0Hz)，3.67(q，1H，J＝7.0Hz)，1.76(d，1H，J＝15.0Hz)，1.54-1.52(m，1H)，1.37(dd，1H，J＝15.0，5.0Hz)，1.15(d，3H，J＝6.5Hz)，1.12(dd，1H，J＝6.0，3.0Hz)，0.92(t，1H，J＝3.3Hz)。

¹³C NMR(125MHz，D ₂O，KOD)δ185.82，182.96，148.01，131.31，129.97，129.78，74.99，73.84，58.78，46.91，38.05，35.02，27.34，27.15。

FTIR(KBr)3366(m)，3072(s)，2886(s)，1696(m)，1611(m)，1560(m)，1455(m)，1377(m)，1278(m)，1202(m)，1188(m)cm ^-1。

C ₁₆H ₁₉NO ₅The analytical calculation value: C, 62.94; H, 6.27; N, 4.59.Measured value: C, 62.70; H, 6.21; N, 4.67.

Preparation example 79

2-[((1R)-and the 1-phenylethyl) amino] (2S, 4S, 6R)-2-formamyl-4-hydroxyl dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester

Though the common original position of title compound is used to prepare 2-[((1R)-the 1-phenylethyl) amino] (2S, 4S, 6R)-4-hydroxyl dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid, but also can separate compound, just, therefore can bring some loss of yield owing to be attended by the hydrolysis of ester in the nitrile hydrolytic process.In the separation, as long as 2-[((1R)-the 1-phenylethyl) amino] (2S, 4S, 6R)-and 2-cyano group-4-hydroxyl dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester is consumed, and the reaction mixture that just makes the nitrile hydrolysis is at CH ₂Cl ₂And H ₂Distribute between the O.Organic layer (the MgSO that is dried ₄) and concentrate after, residuum with EtOAc/ hexane (2: 1)-EtOAc wash-out purifying, obtains title compound through silica gel column chromatography, is white foam shape thing.

[α] ²⁵ _D+61.3(c 1.20，CHCl ₃)。

500MHz ¹HNMR(CDCl ₃)δ 7.32-7.20(m，5H)，7.19(br d，1H，J＝4.0Hz)，5.49(br d，1H，J＝4.0Hz)，4.88(d，1H，J＝11.5Hz)，4.24(dd，1H，J＝11.5，6.0Hz)，4.06-4.00(m，2H)，3.77(q，1H，J＝7.0Hz)，2.21(d，1H，J＝15.0Hz)，2.18-2.15(m，2H)，1.71(br s，1H)，1.54(dd，1H，J＝14.5，6.0 Hz)，1.38，(d，3H，J＝6.5Hz)，1.32(t，1H，J＝3.3Hz)，1.24(t，3H，J＝7.0Hz)。

¹³C NMR(125 MHz，CDCl ₃)δ 180.42，171.47，146.05，128.97，127.43，126.48，73.16，70.76，61.08，56.00，42.82，35.97，35.67，26.13，21.53，14.34。

FTIR(CHCl ₃)3441(m)，3345(m)，2975(w)，1725(s)，1665(s)，1288，1186(m)cm ^-1。

C ₁₈H ₂₄N ₂O ₄The analytical calculation value: C, 65.04; H, 7.28; N, 8.43.Measured value: C, 65.41; H, 7.58; N, 8.32.

Preparation example 80

2-[((1R)-and the 1-phenylethyl) amino] (2S, 4S, 6R)-2-(ethoxy carbonyl)-4-hydroxyl dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester

Under the room temperature, through the 2-[((1R of dropping funnel in 48mL ethanol)-the 1-phenylethyl) amino] (2S, 4S, 6R))-4-hydroxyl dicyclo [3.1.0] hexane-6-carboxylic acid (4g, (11.2mL 157mmol), remains on gentle reflux state so that react 13mmol) to add Acetyl Chloride 98Min. in the suspension.Reflux down gained solution restir 16 hours, be cooled to room temperature after, vacuum concentration is a solid residue.Slowly handle solid with the sodium bicarbonate in the 100mL water (6.6g) solution, use ethyl acetate (2 * 100mL) washings afterwards.Organism drying (the MgSO that merges ₄), filter and vacuum concentration, obtain 4.7g (99%) title compound, it is a solid.Through silica gel column chromatography, use CH ₂Cl ₂/ MeOH (95: 5) wash-out is afterwards again by Et ₂Crystallize out among the O, obtain the analytical pure sample of title compound, be white solid.

[α] ²⁵ _D+52.5(c 1.30，CHCl ₃)。

mp 73-74℃。

500MHz ¹H NMR(CDCl ₃)δ 7.29-7.14(m，5H)，4.25(dq，1H，11.0，7.0Hz)，4.18(dd，1H，J＝9.5，5.5Hz)，4.10(dq，1H，J＝11.0，7.0Hz)，3.92(dq，1H，J＝11.0，7.0Hz)3.82(dq，1H，J＝11.0Hz，7.0Hz)，3.67(q，1H，J＝7.0Hz)，2.73(d，1H，J＝9.5Hz)，2.15-2.12(m，2H)，2.01-1.99(m，1H)，1.89(dd，1H，J＝6.0，3.0Hz)，1.61(dd，1H，J＝15.0，6.0Hz)，1.36(t，1H，J＝3.5Hz)，1.33-1.30(m，6H)，1.18(t，3H，J＝7.0Hz)。

¹³C NMR(125 MHz，CDCl ₃)0δ178.11，171.59，146.32，128.41，127.07，126.85，73.33，70.15，62.07，60.75，56.66，44.72，36.78，33.61，26.24，20.07，14.37，14.23。

FTIR(KBr)3492(s)，3303(m)，3055(w)，2981(w)，2896(w)，1722(s)，1705(s)，1289(m)，1251(m)，1177(m)cm ^-1。

C ₂₀H ₂₇NO ₅The analytical calculation value: C, 66.46; H, 7.52; N, 3.88.Measured value: C, 66.42; H, 7.44; N, 3.92.

Preparation example 81

2-[((1R)-and the 1-phenylethyl) amino] (2S, 4R, 6R)-2-(ethoxy carbonyl)-4-fluorine dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester

Under-20 ℃, with 15 fens clock times, to CH ₂Cl ₂2-[((1R (690mL))-and the 1-phenylethyl) amino] (2S, 4S, 6R))-(59.0g 0.163mol) adds Deoxo-Fluor  (45.1mL to 2-(ethoxy carbonyl)-4-hydroxyl dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester in the solution, 0.245mol), keep temperature to be-15--20 ℃.Stirred the mixture under this temperature 20 minutes, 0 ℃ of following restir 15 minutes slowly adds 15% Na afterwards ₂CO ₃The aqueous solution (650mL), during keep temperature to be lower than 10 ℃.Separate and be equipped with layer, and use CH ₂Cl ₂(150mL) reextraction water layer.Blended organic layer drying (Na ₂SO ₄) and the concentrated brown oil (73g) that becomes.On layer of silica gel,, obtain title compound, be yellow oil (49.7g, 84%) with EtOAc/ heptane (1: 6) wash-out purifying.

[α] ²⁵ _D+36.2(c 1.30，CHCl ₃)。

500MHz ¹H NMR(CDCl ₃)δ7.29-7.14(m，5H)，5.22(ddt，1H，J＝8.0，4.5Hz，JHF＝56.0Hz)，4.16(dq，1H，J＝11.0，7.0Hz)，4.05(dq，1H，11.0，7.0Hz)，3.96(dq，1H，10.5，7.0Hz)，3.85(dq，10.5，7.0Hz)，3.66(q，1H，6.5Hz)，2.45(dd，1H，J＝14.0，8.0Hz)，2.16-2.12(m，1H)，1.95(t，1H，J＝3.5Hz)，1.81(dt，1H，J＝3.5Hz，J _HF＝3.5Hz)，1.51(ddd，1H，J＝14.0，8.0Hz，J _HF＝22.0Hz)，1.32(d，3H，J＝6.5Hz)，1.27(t，3H，J＝7.0Hz)，1.21(t，3H，J＝7.0Hz)。

¹³C NMR(125MHz，CDCl ₃)δ 175.29，171.66，146.21，128.45，127.03，126.90，92.65(d，J _CF＝182 Hz)，68.68(d，J _CF＝4.9 Hz)，61.70，60.92，56.13，38.60(d，J _CF＝23.0Hz)，33.07(d，J _CF＝7.6Hz)，32.23(d，J _CF＝22.0Hz)，26.26，20.22(d，J _CF＝3.9Hz)，14.41，14.24。

FTIR(CHCl ₃)3028(w)，2983(w)，1724(s)，1705(s)，1293(m)，1242(m)，1190(m)，1037(m)，1013(m)cm ^-1。

C ₂₀H ₂₆FN0 ₄The analytical calculation value: C, 66.10; H, 7.21; N, 3.85.Measured value: C, 66.02; H, 7.00; N, 3.95.

Preparation example 82

1R, 2S, 4R, 5R, 6R-2-amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate

With 2-[((1R)-the 1-phenylethyl) amino] (2S, 4S, 6R))-2-(ethoxy carbonyl)-4-fluorine dicyclo [3.1.0] hexane-6-carboxylic acid, ethyl ester (68.4g, 0.188mol), dense HCl (15.7mL, 0.188mol) and 10%Pd/C (exsiccant, 13.7g) mixture in EtOH (400mL) was placed 18 hours down in nitrogen atmosphere (50psi).Filter out catalyzer, distill out overhead product, obtain title compound, for the near-white foam (59.2g, 106% since EtOH impurity calibrated be 97%).By crystallizing out among the EtOH/MTBE, obtain the analytical pure sample of title compound, be white solid.

[α] ²⁵ _D+55.6(c 1.17，CHCl ₃)。

mp 86-88℃。

500MHz ¹H NMR(CDCl ₃)δ9.20(br s，2H)，5.50(ddt，1H，J＝8.0，4.5Hz，J _HF-56.0Hz)，4.31(q，1H，J＝7.0Hz)，4.20-4.07(m，3H)，2.88(t，1H，J＝3.0Hz)，2.71(dd，1H，J＝14.5，8.0Hz)，2.48-2.43(m，2H)，2.16(ddd，1H，J＝14.5，7.5Hz，J _HF＝22.0Hz)，1.34(t，3H，J＝7.0Hz)，1.25(t，3H，J＝7.0Hz)

¹³C NMR(125 MHz，CDCl ₃)δ171.12，169.41，91.94(d，J _CF＝189Hz)，63.85，63.66(d，J _CF＝3.8Hz)，61.73，34.55(d，J _CF＝26.4Hz)，31.58(d，J _CF＝7.8Hz)，30.80(d，J _CF＝-24.1Hz)，20.22，14.31，14.21。

FTIR(KBr)3353(m)，3173(w)，2745(m)，1729(s)，1547(m)，1294(m)，1269(m)，1195(m)，1011(m)cm ^-1。

C ₁₂H ₁₈FNO ₄：C，48.74；H，6.48；N，4.74。Measured value: C, 48.80; H, 6.41; N, 4.76.

Preparation example 83

1R, 2S, 4R, 5R, 6R-2-amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid

With 3N NaOH (251mL, 0.753mol) solution slowly adds to 1R, 2S, 4R, 5R, 6R-2-amino-4-fluorine dicyclo [3.1.0] hexane-2 in the 6-dicarboxylate hydrochlorate (the thick material of 59.2g, 0.188mol theoretical value), keeps temperature to be lower than 26 ℃.Mixture stirred after 10 minutes becomes homogeneous phase.Stirred the mixture under the room temperature 1.25 hours, and with dense HCl pH slowly reduced to pH2.8 afterwards, during to keep temperature be 20-26 ℃.PH is 2.8 times, and mixture begins crystallization, and stirred suspension is 10 minutes under this pH, with dense HCl pH is reduced to 2.1 afterwards.Behind the restir 15 minutes, add i-PrOH (67mL) and suspension is cooled to 0 ℃, stirred 2 hours.Collect solid, with the cold H of 37mL ₂O/i-PrOH (4: 1) washing.With the solid vacuum-drying of collecting 18 hours, obtain title compound under 40 ℃, be white solid (33.1g, 87%, calculate) by the initial substance of preparation example 23.

Preparation example 84

1R, 2S, 4R, 5R, 6R-2-amino-4-fluorine dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid is slurries again

Stir down, with 1 hour, with H ₂1R among the O (165mL), 2S, 4R, 5R, 6R-2-amino-4-fluorine dicyclo [3.1.0] hexane-2, (33.0g, 0.162mmol) suspension is warming up to 89 ℃ to the 6-dicarboxylic acid, and adds i-PrOH (41mL).Reflux then (83 ℃) stirred 5 minutes, be cooled to room temperature again and stirred 4 hours.Collect product, use i-PrOH/H ₂(1: 4,40mL) and i-PrOH (25mL) washing, 40 ℃ of following vacuum-drying 18 hours obtained title compound to O, is white solid (30.6g, 93%).

Preparation example 85

1R, 2S, 4R, 5R, 6R-2-amino-4-fluorine dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate

Under the room temperature, with 20 minutes, the 1R in the absolute dehydrated alcohol of 202mL, 2S, 4R, 5R, 6R-2-amino-4-fluorine dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid (14.45g, 71.12mL) thionyl chloride in the slurries (26mL, 356mol).Slurries are heated to reflux and stirred 3 hours, be cooled to ambient temperature overnight afterwards.With the gained solution for vacuum concentration is residuum, through the dilution of 136mL ethyl acetate, handles 15 minutes with 306mL 10% aqueous sodium carbonate while rotate by hand, and making final pH is 0.Separate each layer, with ethyl acetate (1 * 136mL) washing water layer.The organic extract that merges is through salt solution (1 * 136mL) washing, dry (MgSO ₄), filtering, vacuum concentration obtains 17.07g (93%) title compound, is white solid.

FDMS：M ⁺+1＝260。

C ₁₂H ₁₈FNO ₄·0.1 H ₂O：C，55.21；H，7.03；N，5.37。Measured value: C, 55.10; H, 6.96; N, 5.22.

m.p.＝64-66℃。

[α] ²⁵ _D=+20 (c=0.96, methyl alcohol), [α] ²⁵ _D=+15 (c=1.21, DMSO).

Preparation example 86

1R, 2S, 4R, 5R, 6R-2-[2 ' S-2 '-(tert-butoxycarbonyl amino) propionyl] amino-4-fluorine is two Ring [3.1.0] hexane-2, the 6-diethyl dicarboxylate

With 15 minutes, (38.62g 204mmol) dripped N-methylmorpholine (22.44mL to the N-Boc-L-L-Ala under-22 ℃, nitrogen atmosphere, in the 396mL methylene dichloride in the solution, 204mmol), drip afterwards isobutyl chlorocarbonate (26.48mL, 204mmol) so that temperature of reaction is no more than-18 ℃.Mixed dilute slurry under-20 ℃ 30 minutes, at this moment with 40 minutes adding 1R, 2S, 4R, 5R, 6R-2-amino-4-fluorine dicyclo [3-1.0] hexane-2, (49.46g, the 191mmol) solution that forms in the 247mL methylene dichloride is so that temperature of reaction is no more than-16 ℃ to the 6-diethyl dicarboxylate.After dripping end, from cooling bath, shift out reactant, at room temperature stirred 70 minutes,, this moment, temperature of reaction reached 15 ℃, and color becomes greenish orange look.With 408mL 1N hydrochloric acid processing reaction thing, stir 5 minutes afterwards, and separate each layer.(1 * 408mL) washs organic layer, and dry (sodium sulfate) filters, and vacuum concentration is a white foam shape thing (88.16g) with saturated sodium bicarbonate aqueous solution.

FDMS：M ⁺+1＝260。

C ₁₂H ₁₈FNO ₄·0.1 H ₂O：C，55.21；H，7.03；N，5.37。Measured value: C, 55.10; H, 6.96; N, 5.22.

m.D.＝64-66℃。

[α] ²⁵ _D＝+20(c＝0.96，MeOH)，[α] ²⁵ _D＝+15(c＝1.21，DMSO)。

Preparation example 87

1R, 2S, 4R, 5R, 6R-2-[2 ' S-2 '-(tert-butoxycarbonyl amino) propionyl] amino-4-fluorine is two Ring [3.1.0] hexane-2, the 6-dicarboxylic acid

Under the room temperature, the 1R in the 238mL tetrahydrofuran (THF), 2S; 4R, 5R, 6R-2-[2 ' S-2 '-(tert-butoxycarbonyl amino) propionyl] amino-4-fluorine dicyclo [3.1.0] hexane-2; (88.16g 191mmol) adds 238mL (477mmol) 2N sodium hydroxide to the 6-diethyl dicarboxylate in the solution.This two-phase mixture of vigorous stirring is 2.5 hours under the room temperature, and react homogeneous phase and carry out this moment.With 238mL t-butyl methyl ether diluted mixture thing, mix afterwards, separate each layer again.Further dilute water layer with 238mL water, remove by filter particulate material.With dense HCl (42.9mL, 515mmol) treatment soln is 30 minutes, optional is crystal seed with the title compound subsequently, and stirs 1 hour.Filter the gained slurries, (2 * 100mL) washings, 45 ℃ of following vacuum-drying 40 hours obtains the 72.2g title compound to water, is white solid.Part solid (69.5g) was stirred 1 hour with 490mL acetone, make blushing solution, after filtration, acetone (2 * 100mL) washings.With the filtrate vacuum concentration is white foam shape thing, and 45 ℃ are descended further vacuum-drying 16 hours, obtain 61.8g (having proofreaied and correct the acetone of 12w%wt/wt) title compound.

Embodiment 1

(1R, 4S, 5S, 6S)-and 4-(the amino propionyl of 2 ' S-) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

With 20 minutes, to (1 R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-propionyl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, and the hydrogen chloride solution in the suspension that 6-dicarboxylic acid (110.0g, 271mmol, preparation example 3) forms in the 563mL ethyl acetate in the adding ethyl acetate (3.7M, 514mL).After suspension stirred 2.5 hours, filter and with ethyl acetate (1 * 200mL, 1 * 115mL) washing leaching cake.46 ℃ of following vacuum-dryings were collected title compound after 18 hours, were white solid (85.77g, 92%).

¹HNMR (300MHz, methyl alcohol-d ₄) δ 4.12 (brd, 1H, J=14.6Hz), 3.94 (q, 1H, J=7.1Hz), 3.52 (ddd, 1H, J=7.0,3.9,0.9Hz), 3.16 (d, 1H, J=14.6Hz), 3.02 (dd, 1H, J=7.0,4.4Hz), 2.49 (t, 1H, J=4.1Hz), 1.52 (d, 3H, J=7.1Hz).

Embodiment 2

(1R, 4S, 5S, 6S)-and 4-(2 ' S-2 '-amino propionyl) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylic acid tosylate

With in the toluene (3mL) (1 R, 4S, 5S, 6S)-4-(2 ' S-tert-butoxycarbonyl amino-propionyl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, (140mg, 0.738mmol) suspension is heated to 75 ℃, and stirs 45 minutes, is cooled to room temperature afterwards, restir 16 hours for 6-dicarboxylic acid (300mg, 0.738mmol, preparation example 3) and toluenesulphonic acids monohydrate.Filtering suspension liquid is with toluene (2 * 1mL) washings.45 ℃ of following vacuum-dryings were collected 307mg (87%) title compound after 1 hour, were white solid.

mp(DSC)233℃。

¹HNMR(500MHz，CD ₃OD)δ 7.70(d，2H，J＝8.5Hz)，7.24(d，2H，J＝8.0Hz)，4.11(d，1H，J＝15Hz)，3.94(q，1H，J＝7.0Hz)，3.53(dd，1H，J＝7.0，4.0Hz)，3.13(dd，1H，J＝14，1.0Hz)，3.02(dd，1H，J＝7.0，4.5Hz)，2.48(t，1H，J＝4.5Hz)，2.37(s，3H)，1.52(d，3H，J＝7.5Hz)。

¹³C NMR(125 MHz，CD ₃OD)δ170.70，170.32，169.80，142.04，140.78，128.79，125.79，60.20，54.73，48.77，42.44，30.84，22.22，20.20，16.09。

Embodiment 3

(1R, 4S, 5S, 6S)-and 4-(2 ' S-amino-3 '-hydroxyl-propionyl)-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

According to general method C, use (1R, 4S, 5S, 6S)-and 4-(3 '-acetoxyl group-2 ' S-tert-butoxycarbonyl amino-propionyl)-amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (380mg, 0.77mmol, preparation example 15) is prepared.

[α] ²³ _D=-19.32 (c=0.52, methyl alcohol).

¹HNMR(400MHz，CD ₃OD)δ2.48(1H，t，J＝4.0Hz)，3.04(1H，dd，J＝4.4，7.3Hz)，3.17(1H，d，J＝14.7Hz)，3.51(1H，dd，J＝4.4，7.0Hz)，3.77(1H，dd，J＝7.0，10.6Hz)，3.94-4.12(3H，m)。

C ₁₀H ₁₄N ₂O ₈S·HCl·H ₂O：C，31.88；H，4.55；N，7.44；Cl，9.41。Measured value: C, 31.53; H, 4.40; N, 7.32; Cl, 9.24.

C ₁₀H ₁₅N ₂O ₈The HRMS calculated value of S, 323.0549.Measured value, 323.0533.

Embodiment 4

(1R, 4S, 5S, 6S)-4-[(tetramethyleneimine-2 ' S-carbonyl)-amino]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

According to general method C, use (1 R, 4S, 5S, 6S)-4-[(1 '-tert-butoxycarbonyl-pyrrolidyl-2 ' S-carbonyl)-amino]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (0.8g, 1.7mmol, preparation example 11) is prepared, and obtains 0.42g (67.0%) title compound.

[α] ²³ _D=-32 (c=1.00, methyl alcohol).

¹H NMR(300MHz，CD ₃OD)δ 1.98-2.12(3H，m)，2.40-2.50(1H，m)，2.52(1H，t，J＝4.4Hz)，2.99(1H，dd，J＝4.4，7.0Hz)，3.2(1H，d，J＝14.7Hz)，3.29-3.42(3H，m)，3.54(1H，m)，4.13(1H，d，J＝15.8Hz)，4.30(1H，dd，J＝6.2，9.2Hz)。

C ₁₂H ₁₇N ₂O ₇The HRMS calculated value of S, 333.0756.Measured value, 333.0740.

Embodiment 5

(1R, 4S, 5S, 6S)-and 4-(2 ' S-amino-4 ' methylthio group-butyryl radicals amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

According to general method C, use (1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-4 '-methylthio group-butyryl radicals amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (0.77g, 1.6mmol, preparation example 12) is prepared, and obtains 0.41g (54.9%) title compound.

[α] ²³ _D＝+4(c＝1.00，MeOH)。

¹H NMR(300MHz，CD ₃OD)δ2.1-2.2(2H，m)，2.13(3H，s)，2.47(1H，t，J＝4.4Hz)，2.58-2.63(2H，m)，3.02(1H，dd，J＝4.0，7.0Hz)，3.12(1H，d，J＝14.7Hz)，3.52(1H，dd，J＝3.3，6.6Hz)，3.98(1H，t，J＝6.2Hz)，4.14(1H，d，J＝14.7Hz)。

C ₁₂H ₁₉N ₂O ₇S ₂The HRMS calculated value, 367.0634.Measured value, 367.0634.

Embodiment 6

(1R, 4S, 5S, 6S)-and 4-[2 ' S-amino-3 '-(1H-indol-3-yl) propionyl amino]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

According to general method C, use (1R, 4S, 5S, 6S)-and 4-[2 ' S-tert-butoxycarbonyl amino-3 '-(1-tert-butoxycarbonyl-1H-indol-3-yl)-propionyl amino]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (0.54g, 0.83mmol, preparation example 13) is prepared, and obtains 0.28g (73.6%) title compound.

[α] ²³ _D＝+7.8(c＝1.02，CH ₃OH)。

¹H NMR(300MHz，CD ₃OD)δ2.47-2.53(1H，m)，3.05-3.18(3H，m)，3.44-3.57(2H，m)，4.13-4.23(2H，m)，7.07-7.41(3H，m)，7.71-7.78(2H，m)，8.22(1H，app d，J＝7.7Hz)。

C ₁₈H ₁₉N ₃O ₇The analytical calculation value of SHCl: C, 47.22; H, 4.40; N, 9.18.Measured value: C, 46.51; H, 3.96; N, 8.54.

MS(ES)m/z 420.1[M-1] ^-。

Embodiment 7

(1R, 4S, 5S, 6S)-and 4-[2 ' S-amino-3 '-(4-hydroxyl-phenyl)-propionyl amino]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

According to general method C, use (1R, 4S, 5S, 6S)-and 4-[2 ' S-tert-butoxycarbonyl amino-3 '-(4-tert-butoxycarbonyl-phenyl)-propionyl amino]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (0.33g, 0.53mmol, preparation example 14) is prepared, and obtains 0.13g (56.4%) title compound.

[α] ²³ _D＝-6(c＝1.00，H ₂O)。

¹HNMR(300MHz，CD ₃OD)δ2.47(1H，t，J＝4.0Hz)，2.87(1H，dd，J＝9.2，14.7Hz)，3.05-3.12(2H，m)，3.23(1H，dd，J＝5.1，14.7Hz)，3.55(1H，dd，J＝4.0，7.0Hz)，4.00(1H，dd，J＝4.8，9.2Hz)，4.13(1H，d，J＝14.7Hz)，6.80(2H，d，J＝8.4Hz)，7.13(2H，d，8.8Hz)。

C ₁₆H ₁₉N ₂O ₈The HRMS analytical calculation value of S, 399.0862.Measured value, 399.0844.

Embodiment 8

(1R, 4S, 5S, 6S)-and 4-(2 ' S-amino-3 '-phenyl-propionyl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

According to general method C, use (1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-3 '-phenyl-propionyl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 5) is prepared.Obtain 0.55g (85%, 1.31mmol) white solid.

[α] ²³ _D＝+4.17(c＝0.48，MeOH)。

¹H NMR(300MHz，CD ₃OD)δ2.45(1H，t，J＝4.0Hz)，2.98(1H，dd，J＝9.2，14.7Hz)，3.06(1H，dd，J＝4.4，7.0Hz)，3.10(1H，d，J＝14.6Hz)，3.33(1H，dd，J＝4.8，13.6 Hz)，3.52(1H，dd，J＝3.3，7.3Hz)，4.08(1H，dd，J＝5.1，8.8Hz)，4.10(1H，d，J＝15.4Hz)，7.31-7.42(5H，m)。

¹³C NMR (300MHz, D ₂O w/1, the 4-dioxane): δ 172.55,171.94,170.01,134.31,130.39,130.27,130.01,129.00,61.38,54.76, S4.37,43.07,37.36,31.57,23.18.

C ₁₆H ₁₈N ₂O ₇The analytical calculation value of S1.5HCl: C, 43.97; H, 4.50; N, 6.41.Measured value: C, 43.59:H, 4.17:N, 6.46.

MS (ES) m/z measured value 383.1[M+H] ⁺

C ₁₆H ₁₉N ₂O ₇S [M+H] ⁺The HRMS calculated value: 383.0913.Measured value: 383.0923.

Embodiment 9

(1R, 4S, 5S, 6S)-and 4-(2 ' S-amino-3 ' S-methyl-pentanoyl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

According to general method C, use (1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-3 ' S-methyl-pentanoyl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 6) is prepared.Obtain 0.43g (80%, 1.12mmol) white solid.

[α] ²³ _D＝+4.08(c＝0.49，MeOH)。

¹HNMR(300MHz，CD ₃OD)δ0.98(3H，t，J＝7.3Hz)，1.07(3H，d，J＝7.0Hz)，1.15-1.24(1H，m)，1.53-1.62(1H，m)，1.95-2.04(1H，m)，2.44(1H，t，J＝4.0Hz)，3.06(1H，dd，J＝4.4，7.0 Hz)，3.14(1H，d，J＝15.0Hz)，3.52(1H，dd，J＝3.7，7.0Hz)，3.73(1H，d，J＝5.1Hz)，4.10(1H，d，J＝14.7Hz)。

C ₁₃H ₂₀N ₂O ₇SHCI0.5H ₂The analytical calculation value of O: C, 39.64; H, 5.63; N, 7.11.Measured value: C, 39.38; H, 5.39; N, 7.04.

MS (ES) m/z measured value 349.1[M+H] ⁺

C ₁₃H ₂₁N ₂O ₇S[M+H] ⁺HRMS (ES) calculated value, 349.1069.Measured value, 349.1086.

Embodiment 10

(1R, 4S, 5S, 6S)-and 4-(2 ' S-amino-3 '-methyl-butyryl radicals amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

According to general method C, use (1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-3 '-methyl-butyryl radicals amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 7) is prepared.Obtain 0.18g (88%, 0.49mmol) HCl salt white solid.

[α] ²³ _D＝+7.84(c＝0.51，MeOH)。

¹H NMR(300MHz，CD ₃OD)δ 1.04(3H，d，J＝6.6Hz)，1.09(3H，d，J＝7.0Hz)，2.22-2.29(1H，m)，2.44(1H，t，J＝4.0Hz)，3.05(1H，dd，J＝4.4，7.0Hz)，3.10(1H，d，J＝14.7Hz)，3.52(1H，dd，J＝4.0，7.3Hz)，3.67(1H，d，J＝5.5Hz)，4.10(1H，d，J＝14.7Hz)。

C ₁₂H ₁₈N ₂O ₇S·HCl·0.4H ₂O：C，38.12；H，5.20；N，7.41。Measured value: C, 37.78; H, 4.90; N, 7.15.

MS (ES) m/z measured value 335.1 [M+H] ⁺.

Embodiment 11

(1R, 4S, 5S, 6S)-and 4-(2 ' S-amino-4 '-methyl-pentanoyl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

According to general method C, use (1R, 4S, 5S, 6S)-and 4-(2 ' S-tert-butoxycarbonyl amino-4 '-methyl-pentanoyl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 8) is prepared.Obtain 0.50g (76%, 1.30mmol) HCl salt white solid.

[α] ²³ _D＝-4.0(c＝0.50，MeOH)。

¹HNMR(300MHz，CD ₃OD)δ1.00(3H，d，J＝5.5Hz)，1.02(3H，d，J＝5.9Hz)，1.62-1.79(3H，m)，2.42(1H，t，J＝4.0Hz)，3.04(1H，dd，J＝4.4，7.3Hz)，3.13(1H，d，J＝15.0Hz)，3.52(1H，dd，J＝3.3，7.0Hz)，3.84-3.89(1H，m)，4.10(1H，d，J＝15.0Hz)。

C ₁₃H ₂₀N ₂O ₇S·HCl·0.3H ₂O：C，40.01；H，5.58；N，7.18。Measured value: C, 39.66; H, 5.57; N, 6.99.

MS (ES) m/z measured value 349.1[M+H] ⁺

Embodiment 12

(1R, 4S, 5S, 6S)-and 4-(2 ' S, 6 '-diamino-caproyl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylic acid dihydrochloride

According to general method C, use (1R, 4S, 5S, 6S)-and 4-(2 ' S, 6 '-two tert-butoxycarbonyl amino-caproyl amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (preparation example 9) is prepared.Obtain 0.56g (86%, 1.28mmol) white solid, it is a dihydrochloride.

[α] ²³ _D＝-4.0(c＝0.50，MeOH)。

¹HNMR(300MHz，CD ₃OD)δ1.48-1.55(2H，m)，1.67-1.74(2H，m)，1.89-1.97(2H，m)，2.47(1H，t，J＝4.0Hz)，2.97(2H，app t，J＝4.0)，3.08(1H，dd，J＝4.4，7.0Hz)，3.20(1H，d，J＝15.0Hz)，3.53(1H，dd，J＝3.7，7.0Hz)，3.93(1H，app.t，J＝6.2Hz)，4.08(1H，d，J＝14.7Hz)。

C ₁₃H ₂₁N ₃O ₇S2HCl0.2H ₂O calculated value: C, 35.49; H, 5.36; N, 9.55. measured value: C, 35.30; H, 5.48; N, 9.42.

MS (ES) m/z measured value 364.1 [M+H] ⁺

Embodiment 13

(1R, 4S, 5S, 6S)-and 4-(2 ' S-amino-4 '-formamyl-butyryl radicals amino)-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate

Under the room temperature, with in 1: 1 mixture of 2.5N LiOH and THF (cumulative volume 6mL) (1R, 4S, 5S, 6S)-4-[2 ' S-tert-butoxycarbonyl amino-4 '-(trityl-formamyl)-butyryl radicals amino]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dimethyl dicarboxylate (0.48g, 0.65mmol, preparation example 10) stirred 4 hours.With 1NHCl mixture is adjusted to pH=2, and uses the ethyl acetate extraction product.Merge all organism, use the salt water washing, MgSO ₄Drying concentrates, obtain 0.46g (1R, 4S, 5S, 6S)-4-[2 ' S-tert-butoxycarbonyl amino-4 '-(trityl-formamyl)-butyryl radicals amino]-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the 6-dicarboxylic acid, it is a white foam shape thing.This diacid is dissolved among the DCM, and (0.28g, (3.70g 32.5mmo1) adds TFA 2.6mmol) and subsequently successively with methyl-phenoxide while stirring under the room temperature.Stirred the gained reaction mixture 2 hours under the room temperature, and vacuum concentration.At Et ₂Grind the gained yellow oil among the O, until forming free-pouring white depositions.TFA is collected in vacuum filtration under the nitrogen protection.Product is dissolved among the 1mL HCl and lyophilize, the HCl product that obtains wishing, obtain 0.16g (62%, 0.40mmol) white solid.

[α] ²³ _D＝+8.0(c＝1.0，H ₂O)。

¹HNMR(300MHz，CD ₃OD)δ2.00-2.08(2H，m)，2.37-2.41(3H，m)，2.93(1H，dd，J＝4.4，7.3 Hz)，3.04(1H，d，J＝14.7Hz)，3.45(1H，dd，J＝3.7，7.0Hz)，3.86(1H，app.t，J＝5.9Hz)，4.05(1H，d，J＝14.7Hz)。C ₁₂H ₁₇N ₃O ₈SHCl2.0H ₂O calculated value: C, 33.07; H, 5.09; N, 9.64,

Measured value: C, 33.37; H, 4.69; N, 9.39.

MS (ES) m/z measured value 363.9[M+H] ⁺

C ₁₂H ₁₈N ₃O ₈S[M+H] ⁺HRMS (ES) calculated value: 364.0815, measured value: 364.0825.

Embodiment 14

(1R, 2S, 4R, 5R, 6R)-2-(2 ' S-amino-propionyl) amino-4-fluorine dicyclo [3.1.0] hexane- 2,6-dicarboxylate hydrochlorate

Under 50 ℃, with (1R, 2S; 4R, 5R, 6R)-2-(2 ' S-2 '-(tert-butoxycarbonyl amino) propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2; (53.0g behind the correction acetone, the slurries that 142mmol) form in 447mL acetone stirred 35 minutes the 6-dicarboxylic acid.Filter blushing solution and get settled solution, use the rinsing of 100mL acetone afterwards.With 5 minutes, 22.1mL (265mmol) concentrated hydrochloric acid is dropped in clarification, the near-white filtrate.Mixture is incubated to 45-50 ℃, and stirred 90 minutes, optional with (1R; 2S, 4R, 5R; 6R)-and 2-(the amino propionyl of 2 ' S-) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate stops heating afterwards and is cooled to room temperature gradually as the crystal seed of mixture.After 2 hours, temperature reaches 25 ℃, with 90 minutes acetone (942mL) is added in the slurries.Restir slurries 16 hours filter afterwards, with acetone (2 * 200mL) washings, 45 ℃ of following vacuum-drying 9 hours, under the room temperature dry 64 hours again, obtain 40.2g (91%) title compound, be white solid.

Recrystallization

50 ℃ of following heating edge are dissolved 1.06g in 0.5mL water and 2.12mL acetone, use other 5.3mL acetone diluted afterwards, and the optional crystal seed that adds.In fuzzy cloud mixture, add other 4.2mL acetone, add crystal seed again after choosing wantonly, turn off heating, and in 1 hour, be cooled to room temperature gradually.In 30 minutes, further dilute the gained slurries, stirred afterwards 15 hours with other 9.5mL acetone.After the filtration, with acetone (2 * 5mL) washings, 45 ℃ of following vacuum-drying 10 hours, under the room temperature dry 60 hours again, obtain 0.905g (85% rate of recovery) title compound, be white solid.

mp(DSC)183℃。

[α] ²⁵ _D+33°(c 1.06，CH ₃OH)。

500MHz ¹H NMR(CD ₃OD)δ5.58-5.42(m，1H)，3.92(q，1H，J＝7.0Hz)，2.96(dd，1H，J＝14，8.0Hz)，2.41-2.39(m，1H)，2.35-2.30(m，1H)，2.10(t，1H，J＝3.0Hz)，1.52(d，3H，J＝7.5Hz)，1.51-1.42(m，1H)。

¹³C NMR (125 MHz, CD ₃OD) δ 173.74,173.62,170.00,93.48 and 92.04 (C-F splits branch), 63.95 with 63.92 (C-F splits branch), 48.80,36.89 and 36.70 (C-F splits branch), 32.97 and 32.91 (C-F splits branch), 30.05 with 29.87 (C-F splits branch), 19.37,16.28.FTIR(DRIFT)3430(w)，3016(s)，1721(s)，1662(s)，1496(s)，1190(m)，1024(m)，637(w)cm ^-1。

Embodiment 15

(1R, 2S, 4R, 5R, 6R)-2-(2 ' S-amino-propionyl) amino-4-fluorine dicyclo [3.1.0] hexane- 2,6-dicarboxylic acid mesylate

Under 50 ℃, with (1R, 2S; 4R, 5R, 6R)-and 2-[2 ' S-(tert-butoxycarbonyl amino) propionyl] amino-4-fluorine dicyclo [3.1.0] hexane-2; the slurries that 6-dicarboxylic acid (proofreading and correct back 1.87g, 4.98mmol, preparation example 18) forms in 16.8mL acetone stirred 15 minutes.Filter blushing solution and get settled solution, use acetone (3 * 1.25mL) rinsings afterwards.Dilute clarifying filtrate with 0.935mL water, be placed in 50 ℃ the heating bath, drip with 0.647mL (9.97mmol) methylsulfonic acid and handle (observing gas emits).Make white slurries after 25 minutes.After stirring 2 hours altogether, with the 5-10 minute other 35.5mL acetone of adding.Stop heating and made slurries be cooled to room temperature gradually in 2 hours, filter afterwards, (2 * 8mL) washings, 45 ℃ of following vacuum-drying 14 hours obtains 1.77g (95%) title compound, is the pale pink solid with acetone.The sample of this material carries out recrystallization by following method: under 50 ℃, the limit heating edge is dissolved 1.65g in 1.16mL water and 4.95mL acetone, uses other 1.65mL acetone diluted afterwards, the optional crystal seed that adds.Stop heating, make mixture be cooled to room temperature gradually.Simultaneously with 40 minutes adding acetone (2.64mL).Again the gained slurries were stirred 3 hours.Filter, with acetone (2 * 6mL) washings, 45 ℃ of following vacuum-drying 6 hours, under the room temperature dry 60 hours again, obtain 1.59g (96% rate of recovery) title compound, be white solid:

mp(DSC)206℃。

[α] ²⁵ _D+30(c1.05，CH ₃OH)。

¹HNMR(500MHz，CD ₃OD)δ5.58-5.42(m，1H)，3.92(q，1H，J＝7.0Hz)，2.96(dd，1H，J＝14，8.0Hz)，2.70(s，3H)，2.41-2.39(m，1H)，2.35-2.30(m，1H)，2.10(t，1H，J＝3.0Hz)，1.52(d，3H，J＝7.5Hz)，1.51-1.42(m，1H)。

¹³C NMR (125MHz, CD ₃OD) δ 173.73,173.61,170.02,93.50 and 92.05 (C-F splits branch), 63.91,48.79,38.30,36.89 and 36.70 (C-F splits branch), 32.97 and 32.91 (C-F splits branch), 30.02 and 29.84 (C-F splits branch), 19.37,16.26.

FTIR(DRIFT)3472(w)，3077(s)，1717(s)，1691(s)，1557(m)，1220(s)，1019(m)，781(m)，563(m)cm ^-1。

C ₁₂H ₁₉FN ₂O ₈The analytical calculation value of S: C, 38.92; H, 5.17; N, 7.56.Measured value: C, 38.96; H, 4.97; N, 7.51.

Embodiment 16

(1R, 2S, 4R, 5R, 6R)-2-(2 ' S-amino-propionyl) amino-4-fluorine dicyclo [3.1.0] hexane- 2,6-dicarboxylic acid esilate

Under 50 ℃, will (1R, 2S, 4R, 5R, 6R)-and 2-[2 ' S-(tert-butoxycarbonyl amino) propionyl] amino-4-fluorine dicyclo [3.1.0] hexane-2, the slurries that 6-dicarboxylic acid (0.2g, 0.534mmol, preparation example 18) forms in 1.8mL acetone stirred 5 minutes.Filter blushing solution and get settled solution, use acetone (1 * 0.4mL) rinsing afterwards.Dilute clarifying filtrate with 0.1mL water, be placed in 50 ℃ the heating bath, drip with 0.124mL (1.07mmol) ethyl sulfonic acid and handle (observing gas emits).Make white slurries after 90 minutes.After stirring 2 hours altogether, with the 5 minutes other 1.8mL acetone of adding.Stop heating and made slurries be cooled to room temperature gradually in 1 hour, restir is 2 hours afterwards.Filter, with acetone (2 * 1mL) washings, 45 ℃ of following vacuum-drying 4 hours, under the room temperature dry 60 hours again, obtain 0.173g (84%) title compound, be white solid.210 ℃ of mp (DSC) (decomposition).

¹HNMR(500MHz，CD ₃OD)δ 5.58-5.42(m，1H)，3.92(q，1H，J＝7.0Hz)，2.96(dd，1H，J＝14，8.0Hz)，2.80(q，2H，7.3Hz)，2.42-2.37(m，1H)，2.35-2.30(m，1H)，2.09(t，1H，J＝3.0 Hz)，1.52(d，3H，J＝7.5Hz)，1.51-1.40(m，1H)，1.30(t，3H，J＝7.5Hz)。

Embodiment 17

(1R, 2S, 4R, 5R, 6R)-2-(2 ' S-amino-propionyl) amino-4-fluorine dicyclo [3.1.0] hexane- 2,6-dicarboxylic acid benzene sulfonate

Under 50 ℃, will (1R, 2S, 4R, 5R, 6R)-2-[2 ' S-(tert-butoxycarbonyl amino) propionyl]-amino-4-fluorine dicyclo [3.1.0] hexane-2, the slurries that 6-dicarboxylic acid (0.402g, 1.07mmol, preparation example 18) forms in 3.6mL acetone stirred 10 minutes.Handle blushing solution with one little spoonful of diatomite, and filter settled solution, use acetone (2 * 0.4mL) rinsings afterwards.Clarifying filtrate is placed in 50 ℃ of heating baths, and (90%, 1.29mmol) Phenylsulfonic acid is dissolved in the solution-treated that forms in the 0.113mL water, uses 0.4mL acetone rinsing (observing gas emits) afterwards with 226mg.Gentle reflux stirred after 4 hours, stopped heating and with 8mL acetone treatment reactant 10 minutes, optional afterwards adding crystal seed.Formed slurries through 1 hour, use the 3.2mL acetone diluted, afterwards restir 15.5 hours at room temperature.Filter, (2 * 10mL) washings, 45 ℃ of following vacuum-drying 24 hours obtains 313mg (62%, proofreaied and correct 10wt% acetone) title compound, is white solid with acetone.

mp(DSC)132℃。

¹H NMR(500MHz，CD ₃OD)δ7.86-7.80(m，2H)，7.46-7.37(m，3H)，5.58-5.42(m，1H)，3.92(q，1H，J＝7.0Hz)，2.96(dd，1H，J＝14，8.0Hz)，2.42-2.37(m，1H)，2.35-2.30(m，2H)，2.09(t，1H，J＝3.0Hz)，1.52(d，3H，J＝7.5 Hz)，1.51-1.40(m，1H)。

Embodiment 18

(1R, 2S, 4R, 5R, 6R)-2-(2 ' S-amino-propionyl) amino-4-fluorine dicyclo [3.1.0] hexane- 2,6-dicarboxylic acid tosylate

Under 50 ℃, with (1R, 2S; 4R, 5R, 6R)-2-[2 ' S-(tert-butoxycarbonyl amino) propionyl]-amino-4-fluorine dicyclo [3.1.0] hexane-2; the slurries that 6-dicarboxylic acid (1.04g correction value, 2.78mmol, preparation example 18) forms in 9.36mL acetone stirred 15 minutes.Handle blushing solution with one little spoonful of diatomite, and filter settled solution, use acetone (1 * 2.08mL, 1 * 1.04mL) rinsing afterwards afterwards.Clarifying filtrate is placed in 50 ℃ of heating baths, is dissolved in the solution-treated that forms in the 0.317mL water, use 0.317mL acetone rinsing (observing gas emits) afterwards with 634mg (3.33mmol) tosic acid monohydrate.Gentle reflux stirred after 4 hours, by removing reactant on the heating bath, used 10.4mL acetone treatment reactant 10 minutes.Optional add crystal seed in clarification, colourless solution, observed throw out through 30 minutes and form, add other 10.4mL acetone through 20 fens clock times this moment.Restir slurries 4 hours filter afterwards, and (2 * 10mL) washings, 45 ℃ of following vacuum-drying 14 hours obtains 995mg (78%, proofreaied and correct 3wt% acetone) title compound, is white solid with acetone.

mp(DSC)155℃。

¹HNMR(500MHz，CD ₃OD)δ 7.70(d，2H，J＝7.5Hz)，7.34(d，2H，J＝8.5Hz)，5.58-5.42(m，1H)，3.92(q，1H，J＝7.0Hz)，2.96(dd，1H，J＝14，8.0Hz)，2.42-2.30(m，2H)，2.24(s，3H)，2.09(t，1H，J＝3.0Hz)，1.52(d，3H，J＝7.5Hz)，1.51-1.40(m，1H)。

Embodiment 19

(1R, 2S, 4R, 5R, 6R)-2-(2 ' S-amino-propionyl) amino-4-fluorine dicyclo [3.1.0] hexane- 2, the 6-dicarboxylic acid

Under 50 ℃; (1R in 1mL water; 2S; 4R, 5R, 6R)-2-(2 ' S-amino-propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2; 6-dicarboxylic acid mesylate (0.5g; 1.35mmol embodiment 15) add 5mL3A ethanol in the solution, add 0.27mL (1.35mmol) 5N aqueous sodium hydroxide solution after the several minutes.Stop heating, should clarify colourless solution with the 2.5mL alcohol dilution, the optional crystal seed that adds further diluted 30 minutes with 7.5mL ethanol again.Stir the gained slurries, be cooled to room temperature, afterwards restir 2 hours at room temperature through 1h.Collect solid, (1 * 10mL) washing afterwards 45 ℃ of following vacuum-dryings 18.5 hours, obtains 0.301g (78% yield has been proofreaied and correct 1.6wt% methanesulfonic sodium and 3wt% ethanol) title compound, is white solid with ethanol.

¹HNMR(500MHz，D ₂O)δ 5.45-5.30(m，1H)，3.88(q，1H，J＝7.0Hz)，2.58(dd，1H，J＝14，8.0Hz)，2.33-2.30(m，1H)，2.27-2.26(m，1H)，1.92(t，1H，J＝3.0 Hz)，1.36(d，3H，J＝7.1Hz)，1.41-1.32(m，1H)。

¹³C NMR (125MHz, D ₂O) δ 177.46,176.92,170.42,94.56 and 93.19 (C-F splits branch), 65.36,49.01,36.75 and 36.57 (C-F splits branch), 33.61 and 33.55 (C-F splits branch), 30.54 and 30.36 (C-F splits branch), 20.27,16.67.

Embodiment 20

(1R, 2S, 4R, 5R, 6R)-2-(2 ' S-amino-propionyl) amino-4-fluorine dicyclo [3.1.0] hexane- 2,6-dicarboxylic acid disodium salt

Under 60 ℃; (1R in 420 μ L methyl alcohol; 2S, 4R, 5R; 6R)-2-(2 ' S-amino-propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2; add sodium acetate (46.5mg in 6-dicarboxylic acid mesylate (embodiment 15 for 70mg, the 0.19mmol) solution; 0.57mmol) the warm solution that in 470 μ L methyl alcohol, forms, and with 230 μ L methanol rinse.Stop heating.Dilute this blushing solution with 280 μ L methyl alcohol while stirring, add crystal seed to help crystallization after choosing wantonly.Through 1 hour the gained slurries are slowly cooled to room temperature, and at room temperature stirred 2 hours.The filtering separation product, with methyl alcohol (2 * 280 μ L) washing, 45 ℃ of following vacuum-drying 15 hours obtains 52.5mg (85% yield has been proofreaied and correct 2.3wt% methanesulfonic sodium and 0.2wt% methyl alcohol) title compound, is white solid.

¹HNMR(500MHz，D ₂O)δ 5.44-5.29(m，1H)，3.89(q，1H，J＝7.0Hz)，2.65(s，3H)，2.56(dd，1H，J＝14，8.0Hz)，2.16-2.13(m，1H)，2.10-2.09(m，1H)，1.74(t，1H，J＝3.1Hz)，1.38(d，3H，J＝7.1Hz)，1.36-1.28(m，1H)。

¹³C NMR (125 MHz, D ₂O) δ 180.00,178.72,170.13,95.40 and 93.99 (C-F splits branch), 65.97,49.06,37.25 and 37.07 (C-F splits branch), 33.01 and 32.94 (C-F splits branch), 29.64 and 29.46 (C-F splits branch), 22.48,16.68.

Embodiment 21

(1S, 2S, 4S, 5R, 6R)-2-(the amino propionyl of 2 ' S-) amino-4-hydroxy-dicyclo [3.1.0] hexane- 2,6-dicarboxylate hydrochlorate

(1S in THF (30mL) and water (30mL); 2S; 4S; 5R; 6R)-4-acetoxyl group-2-[2 ' S-(tert.-butoxy) carbonylamino propionyl]-amino dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (0.600g, 1.28mmol; preparation example 21) add in the solution lithium hydroxide monohydrate (0.535g, 12.8mmol).Stir under the room temperature after 3 hours, the dilute with water reactant is with ethyl acetate (2 * 25mL) washings.With 1N HCl water layer is acidified to pH1, with ethyl acetate (3 * 30mL) extractions.The organic extract that merges is dissolved among the 1N HCl in the ethyl acetate (60mL) after concentrating, and stirs 16 hours under the nitrogen atmosphere of room temperature afterwards.Solvent removed in vacuo obtains 400mg (99%) product, is white solid.

LCMS:m/z 273[M+H] ⁺@R _T0.20 minute.

¹HNMR (CD ₃OD) ^*: 4.13 (1H, d, 5.9Hz), 3.80 (1H, q, 6.7Hz), 2.41-2.31 (2H, m), 1.93 (1H, dd, 6.0Hz, 2.7Hz), 1.62 (1H, dd, 5.9Hz, 15.5Hz), 1.44 (1H, t, 3.0Hz), 1.38 (3H, d, 6.8Hz); ^*The unobservable exchangeable protons number of N.B.NMR is 6.

Embodiment 22

(1S, 2S, 4S, 5R, 6R)-2-(2 '-amino-acetylamino-4-hydroxyl-dicyclo [3.1.0] hexane- 2,6-dicarboxylate hydrochlorate

Except that remove the tert-butoxycarbonyl blocking group with the 4M HCl in the dioxane; according to general method C; use (1S; 2S, 4S, 5R; 6R)-2-(2 '-tert-butoxycarbonyl amino-kharophen)-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (300mg, 0.84mmol, preparation example 23) is prepared.Yield 156mg (63%).

[α] ²³ _D＝-36(c＝0.5，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ1.56(1H，t，J＝2.9Hz)，1.74(1H，dd，J＝5.8，15.4Hz)，2.12(1H，m)，2.48(1H，d，J＝15.4Hz)，2.61(1H，m)，3.62(2H，s)，4.32(1H，d，J＝5.49Hz)。

C ₁₀H ₁₄N ₂0 ₆·1.3HCl·H ₂O：C，37.11；H，5.39；N，8.66。Measured value: C, 37.36; H, 4.99; N, 8.30.

MS(ES)m/z 258.8[M+H] ⁺。

Embodiment 23

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-amino-3 '-methyl-butyryl radicals amino)-4-hydroxyl dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate

Except that remove the tert-butoxycarbonyl blocking group with the 4M HCl in the dioxane; according to general method C; use (1S; 2S, 4S, 5R; 6R)-2-(2 ' S-tert-butoxycarbonyl amino-3 '-methyl-butyryl radicals amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (480mg, 1.19mmol, preparation example 24) is prepared.Yield 307mg (76%).

[α] ²³ _D＝+8.33(c＝0.48，MeOH)。

¹H NMR(400MHz，CD ₃OD)δ1.07(3H，d，J＝6.8Hz)，1.11(3H，d，J＝7.3Hz)，1.55(1H，t，J＝2.9Hz)，1.76(1H，dd，J＝5.8，15.6Hz)，2.14(1H，dd，J＝3.4，5.8Hz)，2.24(1H，m)，2.50(1H，d，J＝15.6Hz)，2.64(1H，dd，J＝2.9，5.8Hz)，3.66(1H，d，J＝5.4Hz)，4.32(1H，d，J＝5.8Hz)。

C ₁₃H ₂₀N ₂O ₆HCl1.1H ₂O analytical calculation value: C, 43.79; H, 6.56; N, 7.86.Measured value: C, 43.77; H, 6.20; N, 7.47.

C ₁₃H ₂₁N ₂O ₆The HRMS calculated value, 301.1400.Measured value, 301.1400.

Embodiment 24

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-amino-4 '-methyl-pentanoyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate

Except that remove the tert-butoxycarbonyl blocking group with the 4M HCl in the dioxane; according to general method C; use (1S; 2S, 4S, 5R; 6R)-2-(2 ' S-tert-butoxycarbonyl amino-4 '-methyl-pentanoyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (460mg, 1.11mmol, preparation example 25) is prepared.Yield 371mg (95%).

[α] ²³ _D＝+4(c＝0.5，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ 1.01(3H，d，J＝5.8Hz)，1.03(3H，d，J＝6.3Hz)，1.54(1H，t，J＝2.9Hz)，1.63-1.82(4H，m)，2.14(1H，dd，J＝2.9，5.8Hz)，2.49(1H，d，J＝15.6Hz)，2.62(1H，dd，J＝2.9，5.8Hz)，3.83-3.86(1H，m)，4.32(1H，d，J＝5.8Hz)。

C ₁₄H ₂₂N ₂O ₆HCl1.4H ₂O calculated value: C, 44.72; H, 6.92; N, 7.45.Measured value: C, 44.52; H, 6.57; N, 7.13.

C ₁₄H ₂₃N ₂0 ₆The HRMS calculated value, 315.1556.Measured value, 315.1569.

Embodiment 25

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-amino-3 ' S-methyl-pentanoyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate

Except that remove the tert-butoxycarbonyl blocking group with the 4M HCl in the dioxane; according to general method C; use (1S; 2S, 4S, 5R; 6R)-2-(2 ' S-tert-butoxycarbonyl amino-3 ' S-methyl-pentanoyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (410mg, 0.99mmol, preparation example 26) is prepared.Yield 330mg (95%).

[α] ²⁵ _D＝+8(c＝0.5，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ 0.99(3H，t，J＝7.3Hz)，1.08(3H，t，J＝7.3Hz)，1.17-1.27(1H，m)，1.53(1H，t，J＝2.9Hz)，1.59-1.65(1H，m)，1.76(1H，dd，J＝5.8，15.6Hz)，1.96-2.00(1H，m)，2.14(1H，dd，J＝2.9，5.8Hz)，2.46(1H，d，J＝15.6 Hz)，2.66(1H，dd，J＝2.9，5.8Hz)，3.70(1H，d，J＝5.4 Hz)，4.31(1H，d，J＝5.8Hz)。

C ₁₄H ₂₂N ₂O ₆1.1HCl1.2H ₂O calculated value: C, 44.71; H, 6.83; N, 7.45.Measured value: C, 44.38; H, 6.51; N, 7.08.

C ₁₄H ₂₃N ₂O ₆The HRMS calculated value, 315.1556.Measured value, 315.1566.

Embodiment 26

(1S, 2S, 4S, 5R, 6R)-2-[2 '-(2-amino-acetylamino)-acetylamino]-the 4-hydroxyl Dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

Handle (1S with the 4M HCl in the excessive dioxane; 2S; 4S, 5R, 6R)-2-[2 '-(2-tert-butoxycarbonyl amino-acetylamino)-acetylamino]-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (360mg; 0.77mmol, preparation example 27), and stirred 40 minutes; vacuum concentration adds ethyl acetate and concentrated.Handle this solid with THF (3M) and 1N LiOH (3.0 equivalent).Stirred 1 hour, and added 1N HCl afterwards until pH=3.The vacuum concentration reactant.Through DOWEX  50WX8-100 ion-exchange resin purification.Yield 188mg (78%).

[α] ²³ _D＝+3.92(c＝0.51，H ₂O)。

¹HNMR(300MHz，D ₂O)δ1.43(1H，t，J＝2.9Hz)，1.53(1H，dd，J＝5.9，15.4Hz)，1.98(1H，m)，2.23-2.31(2H，m)，3.70(2H，s)，3.82(2H，app d，J＝1.1Hz)，4.16(1H，d，J＝5.9Hz)。

C ₁₂H ₁₈N ₃O ₇The HRMS calculated value, 316.1145.Measured value, 316.1123.

Embodiment 27

(1S, 2S, 4S, 5R, 6R)-2-[2 '-(2S-amino-propionyl amino)-acetylamino]-the 4-hydroxyl- Dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

According to the general method of general introduction among the embodiment 26, use (1S, 2S; 4S, 5R, 6R)-2-[2 '-(2S-tert-butoxycarbonyl amino-propionyl amino)-acetylamino]-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (410mg, 0.84mmol, preparation example 28).Yield 200mg (72%).

[α] ²³ _D＝+23.53(c＝0.51，H ₂O)。

¹H NMR(300MHz，D ₂O)δ1.36(3H，d，J＝7.0Hz)，1.37(1H，t，J＝3.3Hz)，1.49(1H，dd，J＝5.9，15.4Hz)，1.91(1H，dd，J＝2.9，5.9Hz)，2.15(1H，dd，J＝2.9，5.9Hz)，2.27(1H，d，J＝15.4Hz)，3.79(2H，s)，3.95(1H，q，J＝7.0Hz)，4.13(1H，d，J＝5.5Hz)。

C ₁₃H ₁₉N ₃O ₇The HRMS calculated value, 330.1301.Measured value, 330.1290.

Embodiment 28

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-amino-3 '-phenyl-propionyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate

According to general method C, use (1S, 2S, 4S, 5R, 6R)-and 2-(2 '-tert-butoxycarbonyl amino-3 '-phenyl-propionyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (400mg, 0.89mmol, preparation example 29) is prepared.Yield 290mg (85%).

[α] ²³ _D＝+3.64(c＝0.55，MeOH)。

¹HNMR(400 MHz，CD ₃OD)δ1.45(1H，t，J＝3.3Hz)，1.60(1H，dd，J＝5.5，15.4Hz)，2.03(1H，dd，J＝2.6，5.9Hz)，2.37(1H，d，J＝15.4Hz)，2.55(1H，dd，J＝2.9，5.9Hz)，2.90(1H，dd，J＝8.4，14.3Hz)，3.22(1H，dd，J＝5.1，14.7Hz)，3.97(1H，dd，J＝5.5，8.8Hz)，4.21(1H，d，J＝5.5Hz)，7.19-7.31(5H，m)。

C ₁₇H ₂₁N ₂O ₆The HRMS calculated value, 349.1400.Measured value, 349.1388.

C ₁₇H ₂₁N ₂O ₆HClH ₂The analytical calculation value of O: C, 50.69; H, 5.76; N, 6.95; Cl, 8.80. measured value: C, 50.66; H, 5.65; N, 6.85; Cl, 8.20.

MS measured value 349.0[M+H] ⁺

Embodiment 29

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-amino-4 '-formamyl-butyryl radicals amino)-4-hydroxyl- Dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate

Preparation title compound: with (1S; 2S; 4S; 5R; 6R)-2-[2 '-tert-butoxycarbonyl amino-4 '-(trityl-formamyl)-butyryl radicals amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (1.18mmol, preparation example 30) is dissolved among the THF (10mL), and handles this solution with 2.5NLiOH (12mL).Stirred solution 3 hours is adjusted to pH=2 with 2.5NHCl afterwards.With ethyl acetate extraction product four times, use anhydrous sodium sulfate drying, vacuum concentration obtains white powder.(0.74mmol) is dissolved in 1 with diacid product, in the 2-ethylene dichloride (3.7mL), and handles with methyl-phenoxide (0.3mL) and trifluoroacetic acid (3.72mL).23 ℃ of following reaction stirred 2.5 hours, vacuum concentration obtains brown oil afterwards.This brown oil is dissolved in the water, uses dichloromethane extraction five times, the vacuum concentration water layer is handled with 1N HCl (0.74mL), and freeze-drying.Handle and freeze-drying once more with another part 1N HCl (2mL), obtain 446mg (quantitatively) title compound.

[α] ²³ _D＝+8.16(c＝0.49，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ1.60(1H，t，J＝3.3Hz)，1.79(1H，dd，J＝5.5，15.4Hz)，2.10-2.17(3H，m)，2.46-2.55(3H，m)，2.60(1H，dd，J＝2.9，5.9Hz)，3.94(1H，t，J＝6.2Hz)，4.33(1H，d，J＝5.5Hz)。MS measured value 330.0[M+H] ⁺, 351.9[M+Na] ⁺

C ₁₃H ₁₉N ₃O ₇The HRMS calculated value, 330.1301.Measured value, 330.1295.

Embodiment 30

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S, 6 '-diamino-caproyl amino)-4-hydroxyl-dicyclo [3.1.0] Hexane-2,6-dicarboxylic acid dihydrochloride

According to general method C, use (1S, 2S, 4S, 5R, 6R)-and 2-(2 ' S, 6 '-two-tert-butoxycarbonyl amino-caproyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (preparation example 31) is prepared.

¹HNMR(400MHz，CD ₃OD)δ1.47-1.59(3H，m)，1.66-1.78(2H，m)，1.84(1H，dd，J＝5.4，15.2Hz)，1.91(2H，m)，2.16(1H，dd，J＝3.4，5.7Hz)，2.45(1H，d，J＝15.7Hz)，2.66(1H，dd，J＝2.9，5.9Hz)，2.97(2H，t，J＝7.3Hz)，3.92(1H，t，J＝6.4Hz)，4.33(1H，d，J＝5.4Hz)。MS measured value 330.0[M+H] ⁺

Embodiment 31

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-amino-3 '-formamyl-propionyl amino)-4-hydroxyl- Dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

Preparation title compound: with (1S; 2S; 4S; 5R; 6R)-2-[2 '-tert-butoxycarbonyl amino-3 '-(trityl-formamyl)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (1.07mmol, preparation example 32) is dissolved among the THF (12mL), and handles this solution with 2.5NLiOH (12mL).Stirred solution 3 hours, and be adjusted to pH=2 with 2.5N HCl.With ethyl acetate extraction product four times, use anhydrous sodium sulfate drying, vacuum concentration obtains white powder.(1.22mmol) is dissolved in 1 with diacid product, in the 2-ethylene dichloride (6mL), and handles with methyl-phenoxide (1mL) and trifluoroacetic acid (6mL).23 ℃ of following reaction stirred 10 hours, vacuum concentration obtains brown oil afterwards.Dilute this brown oil with ether, product crystallizes out, and is white solid.Filter, and wash with a large amount of ether.Solid is dissolved in the minimal amount of water, handles with 0.5N HCl (5mL), and freeze-drying 3 times, 386mg (90%) title compound obtained.

[α] ²³ _D＝0(c＝0.5，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ1.59(1H，t，3.4Hz)，1.76(1H，dd，J＝5.9，16.1Hz)，2.11(1H，dd，J＝2.9，5.9Hz)，2.53(1H，d，J＝16.1Hz)，2.55(1H，dd，J＝2.9，5.9Hz)，2.74(1H，dd，J＝9.8，17.1Hz)，2.95(1H，dd，J＝3.9，17.1Hz)，4.19(1H，dd，J＝3.9，9.8Hz)，4.32(1H，d，J＝5.9Hz)。

MS measured value 316.0[M+H] ⁺, 337.9[M+Na] ⁺

C ₁₂H ₁₆N ₃O ₇The HRMS calculated value of Na, 338.0964.Measured value, 338.0953.

Embodiment 32

(1S, 2S, 4S, 5R, 6R)-2-[2 ' S-amino-3 '-(1H-indol-3-yl)-propionyl amino]-the 4-hydroxyl Base-dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

The preparation title compound: with 2-[2 '-tert-butoxycarbonyl amino-3 '-(1 '-tert-butoxycarbonyl-1H-indoles-3 '-yl)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (0.93mmol; preparation example 33) is dissolved among the THF (11 mL), and handles this solution with 2.5N LiOH (11mL).Stirred solution 3 hours is adjusted to pH=2 with 2.5N HCl afterwards.With ethyl acetate extraction product four times, use anhydrous sodium sulfate drying, vacuum concentration obtains white powder.Diacid product (0.78mmol) is dissolved among the 4M HCl in the dioxane (20mL), and 23 ℃ of following reaction stirred 3 hours.Concentrated reaction mixture is with methylene dichloride dilution, vacuum concentration again.Crude product is dissolved in the methyl alcohol, coats radial chromatography (silica gel) plate, Rotary drying is with MeOH (10%)/NH ₄OH (1%)/CHCl ₃-MeOH (60%)/NH ₄OH (2%)/CHCl ₃Wash-out.Be dissolved in the water product and freeze-drying twice, obtain 139mg (46%) title compound.

[α] ²³ _D＝+19.23(c＝0.52，MeOH)。

¹HNMR(400 MHz，CD ₃OD)δ1.43(1H，t，J＝2.9Hz)，1.57(1H，dd，J＝5.9，15.2Hz)，2.05(1H，dd，J＝3.4，6.4Hz)，2.41(1H，d，J＝14.7Hz)，2.46(1H，dd，J＝2.4，5.9Hz)，3.15(1H，dd，J＝5.4，14.7Hz)，3.49(1H，dd，J＝5.4，14.7Hz)，4.05(1H，dd，J＝5.4，9.3Hz)，4.17(1H，d，J＝5.4Hz)，7.09(2H，m)，7.27(1H，s)，7.37(1H，d，J＝8.3Hz)，7.75(1H，d，J＝7.8Hz)。

MS measured value 388.0[M+H] ⁺, 409.9 [M+Na] ⁺

C ₁₉H ₂₁N ₃O ₆The HRMS calculated value, 388.1508.Measured value, 388.1502.

Embodiment 33

(1S, 2S, 4S, 5R, 6R)-4-hydroxyl-2-[(pyrrolidyl-2 ' S-carbonyl)-amino]-dicyclo [3.1.0] Hexane-2,6-dicarboxylate hydrochlorate

Except that remove the tert-butoxycarbonyl blocking group with the 4M HCl in the dioxane; according to general method C; use (1S; 2S; 4S; 5R, 6R)-2-(1 '-tert-butoxycarbonyl-pyrrolidyl-2 ' S-carbonyl)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (preparation example 34) is prepared.

[α] ²³ _D＝-40(c＝0.5，MeOH)。

¹H NMR(400 MHz，CD ₃OD)δ1.59(1H，t，J＝3Hz)，1.78(1H，dd，J＝5.9，15.3Hz)，2.06(4H，m)，2.42(1H，m)，2.59(2H，m)，3.35(m，3H)，4.27(1H，m)，4.32(1H，d，J＝5.9 Hz)。

C ₁₃H ₁₉N ₂O ₆The HRMS calculated value, 299.1243.Measured value, 299.1242.C ₁₃H ₁₈N ₂O ₆HClH ₂O calculated value: C, 44.26; H, 6.00; N, 7.94. measured value: C, 44.13; H, 5.78; N, 7.62.

Embodiment 34

(1S, 2S, 4S, 5R, 6R)-2-[2 ' S-amino-3 '-(4-hydroxyl-phenyl)-propionyl amino]-the 4-hydroxyl Base-dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate

Except that remove the tert-butoxycarbonyl blocking group with the 4M HCl in the dioxane; according to general method C; use (1S; 2S; 4S; 5R, 6R)-2-[2 ' S-tert-butoxycarbonyl amino-3 '-(4-tert.-butoxy carbonyl oxygen base-phenyl)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (preparation example 35) is prepared.Yield 199mg (50%).

[α] ²³ _D＝+8(c＝0.5，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ 1.48(1H，t，J＝3Hz)，1.62(1H，dd，J＝5.9，15.3Hz)，2.03(1H，m)，2.38(1H，d，J＝15.3 Hz)，2.55(1H，m)，2.8(1H，dd，J＝8.9，14.4Hz)，3.12(1H，dd，J＝5.4，14.4Hz)，3.90(1H，q，J＝5.4Hz)，4.21(1H，d，J＝5.9Hz)，6.7(2H，d，J＝8.4Hz)，7.95(2H，d，J＝8.4Hz)。

C ₁₇H ₂₁N ₂O ₇The HRMS calculated value, 365.1349.Measured value, 365.1374.

C ₁₇H ₂₀N ₂O ₇·1.1HCl·1.1H ₂O：C，48.12；H，5.54；N，6.60。Measured value: C, 47.89; H, 5.37; N, 6.50.

Embodiment 35

(1S, 2S, 4S, 5R, 6R)-2-(2 ' S-amino-4 '-methylthio group-butyryl radicals amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-dicarboxylic acid di-ammonium salts

Except that remove the tert-butoxycarbonyl blocking group with the 4M HCl in the dioxane; according to general method C; use (1S; 2S; 4S; 5R, 6R)-2-(2 ' S-tert-butoxycarbonyl amino-4 '-methylthio group-butyryl radicals amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (preparation example 36) is prepared.With MeOH with final product point on the 2mm chromatosheet, use 50/49.5/0.5CHCl ₃/ MeOH/NH ₄The OH wash-out.In the elution process with NH ₄The OH amount increases to 1%, obtains zwitterionic compound.Yield 136mg (37%).

[α] ²³ _D＝+28(c＝1.0，MeOH)。

¹HNMR(400MHz，CD ₃OD)δ1.63(1H，dd，J＝6.0，15.6Hz)，2.03(2H，m)，2.12(3H，s)，2.18(1H，m)，2.35(1H，m)，2.49(1H，d，J＝15.6Hz)，2.65(2H，t，J＝8.4Hz)，3.84(1H，t，J＝6.0Hz)，4.19(1H，d，J＝6.0Hz)。

C ₁₃H ₂₁N ₂O ₆The HRMS calculated value of S, 333.1120.Measured value, 333.1105.

Embodiment 36

(1S, 2S, 4S, 5R, 6R)-2-[2 ' S-(2S-amino-propionyl amino)-propionyl amino]-the 4-hydroxyl Base-dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

With (the 1S among the 4N HCl in the 1ml dioxane; 2S; 4S; 5R; 6R)-2-[2 ' S-(2S-tert-butoxycarbonyl amino-propionyl amino)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (0.045g, 0.1mmol; preparation example 39) stirred 40 minutes, afterwards the vacuum concentration reactant.Thick material among THF (5mL) and the 0.5N LiOH (0.35mmol) was stirred 1 hour.Be adjusted to pH=2 with 0.5N HCl, and the concentration response thing.Use cation-exchange chromatography (Dowex  50X8-100; With 10% pyridine/H ₂The O wash-out) purifying crude product obtains 19mg (53%) title compound.

¹HNMR(300MHz，D ₂O)δ1.23(3H，d，J＝7.3Hz)，1.36(3H，d，J＝7.3Hz)，1.40-145(1H，m)，1.50(1H，dd，J＝5.5，15.4Hz)，1.90-1.96(1H，m)，2.20(1H，dd，J＝2.6，5.9Hz)，2.29(1H，d，J＝15.4Hz)，3.90(1H，q，J＝7.3 Hz)，4.15-4.22(2H，m)。

MS(ES)m/z 342.1[M-1] ^-。

C ₁₄H ₂₂N ₃O ₇The HRMS calculated value, 344.1458.Measured value, 344.1457.

Embodiment 37

(1S, 2S, 4S, 5R, 6R)-2-[2 ' S-(2-amino-acetylamino)-propionyl amino]-the 4-hydroxyl- Dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

With (1 S among THF (5mL) and the 1 N LiOH (1.0mmol); 2S; 4S; 5R; 6R)-2-[2 ' S-(2-tert-butoxycarbonyl amino-acetylamino)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2; 6-diethyl dicarboxylate (0.12g, 0.25mmol, preparation example 40) stirred 1 hour.With 1NHCl mixture is adjusted to pH=2 and concentrated.In saturated HCl 0 ℃ contain (g) ethyl acetate solution, stir crude product.Use N ₂Purging is removed excessive HCl (g) and concentration response thing.Use cation-exchange chromatography (Dowex  50X8-100; With 10% pyridine/H ₂The O wash-out) purifying crude product obtains 0.06g (72.9%) title compound.

[α] ²³ _D＝-42.11(c＝0.57，H ₂O)。

¹H NMR(300MHz，D ₂O)δ1.21(3H，d，J＝7.3Hz)，1.39(1H，m)，1.50(1H，dd，J＝5.7，15.8Hz)，1.91(1H，m)，2.15(1H，dd，J＝2.6，5.9Hz)，2.28(1H，d，J＝15.0Hz)，3.65(2H，s)，4.14(1H，app d，J＝5.9Hz)，4.20(1H，app q，J＝7.3Hz)。

C ₁₃H ₂₀N ₃O ₇The HRMS calculated value, 300.1301.Measured value, 330.1299.

Embodiment 38

(1S, 2S, 4S, 5R, 6R)-2-[2 ' S-(2S-amino-4-methyl-pentanoyl amino)-propionyl ammonia Base]-4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

According to general method C, use (1S, 2S, 4S, 5R, 6R)-2-[2 '-(2S-tert-butoxycarbonyl amino-4-methyl-pentanoyl amino)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (preparation example 41) is prepared.Obtain 0.21g (57%, 0.50mmol) white solid.

[α] ²³ _D＝-3.64(c＝0.55，MeOH)。

¹HNMR(300MHz，CD ₃OD)δ 0.89(3H，d，J＝4.4Hz)，0.91(3H，d，J＝4.0Hz)，1.29(3H，d，J＝7.0Hz)，1.52(1H，t，J＝3.3Hz)，1.55-1.67(4H，m)，1.97(1H，dd，J＝2.9，5.9Hz)，2.38(1H，dd，J＝2.9，5.9Hz)，2.45(1H，d，J＝15.4Hz)，3.75-3.80(1H，m)，4.18g(1H，d，J＝5.9Hz)，4.33(1H，app.q，J＝7.0 Hz)，

C ₁₇H ₂₇N ₃O ₇1.0HCl1.6H ₂0 analytical calculation value: C, 45.30; H, 6.98; N, 9.32; Cl, 7.87.

Measured value: C, 44.95; H, 6.54; N, 9.12; Cl, 7.53.

C ₁₇H ₂₈N ₃O ₇[M+H] ⁺HRMS (ES) calculated value, 386.1927.Measured value, 386.1911.

Embodiment 39

(1S, 2S, 4S, 5R, 6R)-2-[2 ' S-(2-amino-3-methyl-butyryl radicals amino)-propionyl amino]- 4-hydroxyl-dicyclo [3.1.0] hexane-2, the 6-dicarboxylic acid

According to-as method C, use (1 S, 2S, 4S, 5R, 6R)-2-[2 '-(2S-tert-butoxycarbonyl amino-3-methyl-butyryl radicals amino)-propionyl amino]-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (preparation example 42) is prepared.Obtain 0.15g (88%, 0.37mmol) white solid.

[α] ²³ _D＝-15.69°(c＝0.51，MeOH)。

¹H NMR(400 MHz，CD ₃OD)δ 0.92(3H，d，J＝7.0Hz)，0.95(3H，d，J＝7.0Hz)，1.28(3H，d，J＝7.0Hz)，1.52(1H，t，J＝2.9Hz)，1.60(1H，dd，J＝5.9，15.4Hz)，1.98(1H，dd，J＝3.3，5.9Hz)，2.03-2.15(1H，m)，2.39(1H，dd，J＝2.9，6.2 Hz)，2.45(1H，d，J＝15.4Hz)，3.56(1H，d，J＝5.5Hz)，4.18(1H，d，J＝5.9Hz)，4.33(1H，app.q，J＝7.0Hz)，8.82(1H，s)。

C ₁₆H ₂₅N ₃O ₇1.0HCl1.5H ₂The analytical calculation value of O: C, 44.19; H, 6.72; N, 9.66; Cl, 8.15.

Measured value: C, 44.32; H, 6.48; N, 9.14; Cl, 7.66.

C ₁₆H ₂₆N ₃O ₇[M+H] ⁺HRMS (ES) calculated value, 372.1771.Measured value, 372.1758.

Embodiment 40

(1R, 2S, 4S, 5R, 6R)-2-(2 ' S-2 '-amino propionyl) amino-4-fluoro-dicyclo [3.1.0] hexane -2,6-dicarboxylic acid mesylate monohydrate

Under the gentle reflux; (1R, 2S, 4S in 295mL acetone; 5R; 6R)-and 2-(2 ' S-2 '-(tert-butoxycarbonyl amino) propionyl) amino-4-fluoro-dicyclo [3.1.0] hexane-2,6-dicarboxylic acid (32.8g, 87.5mmol; preparation example 52) drips 16mL water in the blushing solution; drip 11.4mL (175mmol) methylsulfonic acid afterwards, obtain clarification, yellow solution, it became slurries after 5 minutes.Stirred 130 minutes, and removed thermal source, added other 295mL acetone through 30 minutes.Slurries are cooled to room temperature, and then stirred 2 hours.Filter slurries, (2 * 82mL) washings, 45 ℃ of following vacuum-drying 16 hours obtains 31.5g (93%) title compound, is white solid with acetone.

This material is recrystallization further.The above-mentioned crude product of 30.5g is mixed with 152.5mL acetone and 35mL water.Be heated to 55 ℃, add water (3.66mL) and promote dissolving fully.With 61mL acetone diluted solution, and the optional crystal seed that adds.Remove thermal source, mixture is cooled off gradually until beginning to form good nucleus.In 70 fens clockwise slurries, add 396mL acetone, then restir 3 hours under the room temperature.Filter, (3 * 91mL) washings, 45 ℃ of following vacuum-dryings several hours (spending the night usually) obtain 27.7g (91% rate of recovery) title compound, are white solid with acetone.mp(DSC)200℃。

[α] ²⁵ _D+34°(c 1.0，CH ₃OH)。

400MHz ¹H NMR(DMSO-d ₆)δ12.76(br s，2H)，9.18(s1H)，8.07(brs，3H)，5.50-5.36(m，1H)，3.87(d，1H，J＝6.8Hz)，2.82(dd，1H，J＝14，8.0Hz)，2.38(s，3H)，2.25(m，2H)，1.96(t，1H，J＝3.0Hz)，1.39(m，1H)，1.37(d，3H，J＝6.8Hz)。

100 MHz ¹³C NMR (DMSO-d ₆) δ 173.1,172.3,169.7,92.8 (d, C-F splits branch), 63.2 (d, C-F splits branch), 48.0,39.7,36.4 (d, C-F splits branch), 32.5 (d, C-F splits branch), 29.3 (d, C-F splits branch), 19.3 (d, C-F splits branch), 16.9.FTIR(KBr)3461(w)，3379(w)，3269(m)，2653(s)，2591(s)，2529(s)，1724(s)，1691(s)，1353(m)，1287(s)，1271(s)，1256(s)，1212(s)，1147(s)，1052(s)，1024(s)，787(m)cm ^-1。

C ₁₂H ₂₁FN ₂O ₉S calculated value: C, 37.11; H, 5.45; N, 7.21. measured value: C, 37.12; H, 5.45; N, 7.16.

Embodiment 41

(1R, 4S, 5S, 6S)-and 4-(2 ' S-4 '-methylthio group-2 '-amino butyryl radicals) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylic acid monohydrate

Will (1R, 4S, 5S, 6S)-and 4-(2 ' S-4 '-methylthio group-2 '-(tert-butoxycarbonyl) amino butyryl radicals) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, the slurries of 6-dicarboxylic acid list sodium salt (110.04g, 225.3mmol, preparation example 55) in acetone (110mL) and water (550mL) are heated to 55 ℃.In the slurries that stir, drip concentrated hydrochloric acid (56mL, 675.8mmol) so that it dissolves gradually.After dripping end, stirred 2 hours down at 55 ℃.Remove thermal source, make solution be cooled to room temperature.Filtering solution is used the 20mL water rinse.(165mL 330mmol), makes pH rise to 1.71, this moment occurs precipitation slowly to add 2N sodium hydroxide in solution.Stirred 10 minutes; Form dilute slurry, pH reduces to 0.98.(62mL 124mmol), makes pH rise to 3.06, stirs afterwards 3 hours, gets final pH 3.24 to add more 2N sodium hydroxide in the gained slurries.Filter slurries, (2 * 165mL) washings, 45 ℃ were descended dry 15 hours water, obtained 73.27g (85% weight yield) title compound, were white solid.

mp(DSC)203℃。

[α] ²⁵ _D+13.4(c1.19，1N HCl)。

500MHz ¹H NMR(D ₂O)δ3.99(t，1H，J＝6.0Hz)，3.93(d，1H，J＝15.0Hz)，3.50(dd，1H，J＝1.0，4.0Hz)，3.12(d，1H，J＝15.0Hz)，2.95(dd，1H，J＝4.0，7.0Hz)，2.48(t，2H，J＝8.0Hz)，2.33(t，1H，J＝4.0)，2.09-1.98(m，5H)。

¹³C NMR(125 MHz，D ₂O)173.50，172.60，169.18，61.66，54.76，52.19，42.55，31.70，30.10，28.09，23.53，14.14。

FTIR(ATR)3558.54(s)，3024.05(s)，2959.87(s)，1748.83(s)，1692.89(s)，1681.99(s)，1617.50(s)，1567.63(s)，1497.65(s)，1314.11(s)，1282.22(s)，1263.26(s)，1239.01(s)，1101.46(s)，884.62(s)，809.95(s)，773.46(s)cm ^-1。

C ₁₂H ₁₈N ₂O ₇S ₂H ₂0 analytical calculation value: C, 37.49; H, 5.24; N, 7.29.Measured value: C, 37.34; H, 5.04; N, 7.15.

Embodiment 42

(1R, 4S, 5S, 6S)-and 4-(2 ' S-4 '-methylthio group-2 '-amino butyryl radicals) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylic acid tosylate

With in the toluene (1180mL) (1R, 4S, 5S, 6S)-4-(2 ' S-4 '-methylthio group-2 '-(tert-butoxycarbonyl) amino butyryl radicals) is amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4, (54g, 278mmol) mixture heating up to 75 ℃ obtains underflow liquid for 6-dicarboxylic acid (118.09g correction value, 253mmol, preparation example 56) and tosic acid monohydrate.Reflux and stirred slurries 165 minutes down.Remove thermal source, make slurries be cooled to room temperature, stir afterwards and spend the night.Filter slurries, (3 * 240mL) washings, 45 ℃ of following vacuum-drying 22 hours obtains 134.92g (98% yield) title compound with toluene.

mp(DSC)255℃。

[α] ²⁵ _D+8.3(c 1.2，CH ₃OH)。

500MHz ¹H NMR(CD ₃OD)δ 7.71(d，2H，J＝8.0Hz)，7.24(d，2H，J＝8.0Hz)，4.14(d，1H，J＝15Hz)，4.00(t，1H，J＝6.0Hz)，3.54(dd，1H，J＝4.0，7.0Hz)，3.13(d，1H，J＝15Hz)，3.01(dd，1H，J＝4.0，7.0Hz)，2.60(t，2H，J＝8.0Hz)，2.49(t，1H，J＝4.0Hz)，2.37(s，3H)，2.19-2.12(m，5H)。

¹³C NMR(125 MHz，CD ₃OD)δ 170.49，169.69，168.99，142.18，140.67，182.73，125.79，60.26，54.76，52.21，42.44，30.90，30.77，27.20，22.33，20.17，13.96。

FTIR(ATR)3091.19(w)，1730.91(s)，1668.22(s)，1563.97(s)，1518.49(s)，1312.69(m)，1247.46(s)，1212.05(s)，1156.33(s)，1123.09(s)，1035.95(s)，1011.36(s)，892.41(s)，814.02(s)，683.69(s)cm ^-1。

Embodiment 43

(1R, 4S, 5S, 6S)-and 4-(2 ' S-4 '-methylthio group-2 '-amino butyryl radicals) amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylic acid mesylate

With in the 13mL propionitrile (1R, 4S, 5S, 6S)-4-(2 ' S-4 '-methylthio group-2 '-(tert-butoxycarbonyl) amino butyryl radicals) is amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylic acid (1.08g, 2.31mmol, preparation example 56) slurries are heated to 85 ℃.In slurries, add entry (540 μ L), drip then methylsulfonic acid (225 μ L, 3.47mmol).Stirred slurries 90 minutes.Remove thermal source and add propionitrile (30mL).Slurries are cooled to room temperature and stirred 90 minutes.Filter, (3 * 2.7mL) washings, 45 ℃ of following dried overnight obtain 1.04g (97%) title compound with propionitrile.mp(DSC)244℃。

[α] ²⁵ _D+10.2(c 1.16，CH ₃OH)。

500MHz ¹H NMR(CD ₃OD)δ 4.16(d，1H，J＝15Hz)，4.00(t，1H，J＝6.0Hz)，3.54(dd，1H，J＝4.0，7.0Hz)，3.15(d，1H，J＝15Hz)，3.01(dd，1H，J＝4.0，7.0Hz)，2.71(s，3H)，2.61(t，2H，J＝8.0)，2.51(t，1H，J＝4.0)，2.20-2.14(m，5H)。

¹³C NMR(125 MHz，CD ₃OD)δ170.50，169.71，169.00，60.27，54.78，52.18，42.43，38.35，30.90，30.78，28.20，22.35，13.96。FTIR(ATR)3055.57(m)，1725.90(s)，1693.60(s)，1527.33(s)，1528.96(s)，1320.89(s)，1176.86(s)，1152.70(s)，1118.55(s)，1051.42(s)，816.49(s)，786.63(s)cm ^-1。

C ₁₂H ₁₈N ₂O ₇S ₂CH ₄O ₃The analytical calculation value of S: C, 33.76; H, 4.79; N, 6.06.Measured value: C, 33.98; H, 4.82; N, 5.98.

Embodiment 44

(1R, 2S, 5R, 6R)-and 2-(the amino propionyl of 2 ' R-) amino-6-fluoro-dicyclo [3.1.0] hexane-2,6- The dicarboxylate hydrochlorate

In the solution of the isomer A of 0.20g (0.47mmol) preparation example 60 in THF (1.2mL), add 1.13mL (2.8mmol) 2.5N LiOH, and at room temperature stir the mixture and spend the night.With the ether washing, 0 ℃ neutralizes with 1N HCl down, and (3 * 3mL) extract with EtOAc.With dry organic layer and the concentrating under reduced pressure that merges of anhydrous MgSO4.Dissolving crude product is contained among the EtOAc of 1NHCl and stirs and spend the night at 3.76ml.Incline and desolvate, with the ether washing, desciccate under the Ar air-flow obtains title compound (83mg, 57% yield), is white solid.

¹HNMR(300MHz，CDCl ₃)：1.28(d，3H，J＝7.1Hz)，1.64-1.69(m，1H)，1.86-2.02(m，1H)，2.11-2.21(m，3H)，2.54(d，1H，J＝7.1Hz)，3.84(q，1H，J＝7.1Hz)。

Embodiment 45

(1S, 2R, 5S, 6S)-and 2-(the amino propionyl of 2 ' R-) amino-6-fluoro-dicyclo [3.1.0] hexane-2,6- The dicarboxylate hydrochlorate

Solution with the isomer B of preparation example 60 is initial substance, makes title compound according to embodiment 44 substantially.

¹HNMR(300MHz，CDCl ₃)：1.30(d，3H，J＝7.1Hz)，1.59-1.65(m，1H)，1.85-2.03(m，1H)，2.05-2.25(m，3H)，2.53(d，1H，J＝6.6Hz)，3.85(q，1H，J＝7.1Hz)。

Embodiment 46

(1S, 2S, 4S, 5R, 6R)-2-(the amino propionyl of 2 ' S-) amino-4-hydroxy-dicyclo [3.1.0] hexane- 2,6-dicarboxylate hydrochlorate

According to general method C, use (1S, 2S, 4S; 5R, 6R)-2-(2 ' S-tert-butoxycarbonyl amino-propionyl amino)-4-hydroxyl-dicyclo [3.1.0] hexane-2,6-diethyl dicarboxylate (0.47g; 1.1mmol, preparation example 37) be prepared, obtain 0.23g (67.7%) title compound.

[α] ²³ _D＝-4.1(c＝0.49，MeOH)。

¹HNMR(CD ₃OD)δ 1.51(3H，d，J＝7.0Hz)，1.56(1H，t，J＝2.9Hz)，1.77(1H，dd，J＝5.8，15.8Hz)，2.1-2.13(1H，m)，2.52(1H，d，J＝16.1Hz)，2.57(1H，dd，J＝2.9，5.9 Hz)，3.88(1H，dd，J＝7.0，14.3Hz)，4.32(1H，d，J＝5.5Hz)。

C ₁₁H ₁₇N ₂O ₆1.1HCl0.9H ₂O analytical calculation value: C, 40.12; H, 5.80; N, 8.53.Measured value: C, 39.85; H, 5.41; N, 8.36.

C ₁₁H ₁₆N ₂O ₆The HRMS calculated value of Na, 295.0906.Measured value, 295.0883.

Drug compound can be estimated by various cell absorption tests corresponding to corresponding parent compound before of the present invention.These tests can provide comparative data, thereby thereby those of ordinary skills can be differentiated to be easy to be absorbed the compound that brings more superior exposure character by cell.Two this samples are described below, comprise Gly-Sar absorption test and Caco-2 test.

The Gly-Sar absorption test

It has been recognized that the plan peptide medicament of some oral administrations absorbs by the peptide haulage system in the intestines.Yang etc., Pharm.Res.16 (9) (1999).Especially, people study suppressor mode and intracellular corresponding identification level thereof that peptidyl absorbs peptide carrier hPepT1 in the intestines.Meredith etc., Eur.J.Biochem., 267,3723-3728 (2000).And as the available strategy of differentiating that improved oral pharmaceutical absorb, people have set about characterizing hPepT1 and have transported absorption mechanism in the intravital amino acid whose intestines.Han etc., Polym.Prepr. (Am.Chem.Soc., Div.Polym.Chem) 40 (1): 259-260 (1999); Sawada etc., J.Pharmacol.Exp.Ther., 291 (2): 705-709 (1999).

U.S. Patent No. 5,849 has been described the method that can be used for measuring The compounds of this invention and hPepT1 carrier affinity level in 525.

For example, the Chinese hamster ovary of the stable transfection of overexpression hPepT1 carrier (CHO) cell can be used for testing The compounds of this invention.Chinese hamster ovary celI is used to monitor the absorption of Gly-Sar, when drug compound exists before of the present invention, the absorption of pair cell greater than do not exist of the present invention before absorption during drug compound, provable in view of the above its agonist activity; If to the absorption of drug compound before of the present invention absorption when not having drug compound before the present invention, then provable its suppresses active.The EC of The compounds of this invention ₅₀Value is usually less than 5mM.

The Caco-2 test

The intestines inner cell that another adhoc approach of measuring the absorption of The compounds of this invention cell is the research people is the peptide transport agent of Caco-2.Adenocarcinoma cell (Memorial sloan-KetteringCancer Center with the people, Rye, NY, and/or ATCCP, Rockville, MD) in improvement Yi Geer (Modified Eagle) medium of the Dulbecco that contains 10% foetal calf serum and 1% minimum non-essential amino acid solution that must medium, go down to posterity, do not add Sodium.alpha.-ketopropionate or antibiotic.These cells do not conform to mycoplasma, and are to use between 28～40 generations in passage number.Measure 5-10 * 10 for effluent ⁴Individual cell has applied in 13-18 days time in the porous ware of collagen protein and has grown out, and changes once in the every 2-3 of medium days.

Under 37 ℃, the use test compound is according to group dish (cluster-tray) (referring to Gazzola etc., Anal.Biochem.115,368-74 (1981)) technical measurement drug absorption.The effluent damping fluid for potassium-containing hydrogen salt not, contain Earle ' the s balanced salt that useful KOH titration is 6.0 25mM Mes to pH, and replace sodium-chlor with Lipotril.With Lipotril the osmolarity of effluent damping fluid is adjusted to 300 ± 5mosmol/kg.Will [ ³H] synanthrin is as the mark that is attached to the extracellular fluid on the cell in the washing process, to measure the time of origin of determining uptake rate.The fresh solution of test compound and dipeptides will prepare every day.Last in experiment, in water, use LC/MS/MS can detect the interior test compound of cell lysates cytolysis.With Smith etc., the method for describing among the Anal.Biochem.150,76-85 (1985) is measured protein.

Measured absorption through 40 minutes.In the linearity region of process fall time, calculate initial absorption speed, use linear regression to estimate above-mentioned time of origin.Calculate percent inhibition based on there not being the contrast uptake rate of measuring under the dipeptides situation.For example this Caco-2 test is referring to Dantzig﹠amp; Bergin, Biochim.Biophys.Acta1027,211-17 (1990).

By exposure situation in the body of rat plasma concentration determination

For exposure situation in the body of studying formula II compound, study the oral dosage contrast of formula I compound and formula II compound afterwards, carried out measuring the research of the plasma concentration of various II compound in rat.Grow up Fischer344 male rat (190-270 gram) by HarlanSprague-Dawley, Cumberland, IN, USA obtains, and cultures its adaptation of 3 angels in the laboratory.The 4th day, with test compound be dissolved in buffered water (1mg/ml=test compound/20mM potassium primary phosphate, pH=2) in, and with the oral dose administration of single 5mg/kg.At 0.5 and 1 hour, or located, pierce through (last time point) through eye socket fistula or heart and collect blood sample at 1 and 3 hour.Plasma sample is before analyzing, prior to-20 ℃ and phenylmethylsulfonyl fluoride (a kind of proteinase inhibitor) existence storage down.With solid-phase extraction method (SAX carrier, methanol/dilute acetic acid) pre-treatment plasma sample and interior mark compound.

Shown in following table 1A and 1B, determine the plasma concentration (ng/ml) of the various II compound of each test with LC/MS/MS, and as 0.5 and 1 hour or, the total concn of 1 and 3 hour sample time point provides.

Table 1AExposure test in the body
		Compound	The rat exposure (ng/ml (1R, 4S, 5S, 6S)-and 4-(the amino propionyl of 2 ' S-) amino-2,2-dioxo-2 λ 6-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate
Embodiment 1	2251ng/ml (according to 10mg/kg p.o.)
		The non-prodrug forms of embodiment 1	1521ng/ml (according to 5mg/kg p.o.) 3981Hg/ml (according to 10mg/kg p.o.)

Table 1BExposure test in the body
		Compound	Rat exposes (ng/ml (1R, 2S, 4R, 5R, 6R)-and 2-(the amino propionyl of 2 ' S-) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate
Embodiment 14	5271ng/ml (according to 5mg/kg p.o.)
		The non-prodrug forms of embodiment 14	1162ng/ml (according to 5mg/kg p.o.) 1342ng/ml (according to 10mg/kg p.o.)

As above show shown in 1A and the 1B, during to the rat oral medication, the plasma concentration of The compounds of this invention and parent compound be comparable.This illustrates that compound of the present invention changes parent compound in vivo into, formula II compound.

Compound of the present invention is first preparation before administration preferably.Therefore, another aspect of the present invention is the pharmaceutical preparation that comprises formula I compound or its pharmaceutically useful salt, pharmaceutically useful carrier, thinner or excipient.Can prepare this pharmaceutical preparation according to method known to a person of ordinary skill in the art.When preparing composition of the present invention, usually that active ingredient and carrier is mixed, or with the carrier dilution or with the carrier sealing, and can place capsule, pouch, paper or other to hold in the bodily form formula.When carrier was used as thinner, it can be solid, semisolid or liquid substance, and it is carrier, excipient or the medium of active ingredient.Composition can be tablet, pill, powder, lozenge, pouch, cartridge bag, elixir, suspension, emulsion, solution, syrup, aerosol, for example contains ointment, soft and hard gelatine capsule, suppository, aseptic parenteral solution and the aseptic packaging powder of 10% active compound that weighs at the most.

Some examples of carrier, excipient and the thinner that is fit to comprise lactose, glucose, sucrose, Sorbitol Powder, mannitol, starch, natural gum, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, Calucium Silicate powder, Microcrystalline Cellulose, polyvinylpyrrolidone, Mierocrystalline cellulose, aqueous syrup, methylcellulose gum, methyl hydroxybenzoate and propyl ester, talcum, Magnesium Stearate and mineral oil.Preparation also can comprise lubricant, wetting agent, emulsifying agent and suspension agent, sanitas, sweetener or seasonings in addition.Composition of the present invention can be made into preparation, like this according to method well known in the art after patient's administration, bring active ingredient fast, lasting or postpone to discharge.

Preferably by measure unit form preparation, every dosage contains the about 500mg active ingredient of the 5mg-that has an appointment to composition, the about 300mg active ingredient of preferably about 25mg-.The term of Shi Yonging " active ingredient " refers to be included in the compound in the formula I compound scope herein.

Term " unit dosage form " refers to be suitable for the physiology discrete unit as human body and other mammiferous single dose, and every unit contains the active substance (it has calculated the result of treatment that can produce hope) and suitable pharmaceutical carrier, thinner or the excipient of predetermined amount.

Claims (27)

1. the compound of a formula I or its pharmaceutically useful salt,

Wherein,

A is H-(Q) _p-;

When occurring at every turn, Q independently is selected from aminoacyl;

P is the integer of 1-10;

X is O, S, SO, SO ₂Or CR ³R ⁴

R ³Be fluorine, X ' OR ⁵, SO ₃H, tetrazolium-5-base, CN, PO ₃R ⁶ ₂, hydroxyl, NO ₂, N ₃, (CH ₂) mCOOR ^5a, (CH ₂) mPO ₃R ^6a ₂, NHCONHR ^5bOr NHSO ₂R ^5c, and R ⁴Be hydrogen; Or R ³And R ⁴Represent fluorine separately; Or R ³And R ⁴Together representative=O ,=NOR ⁷,=CR ⁸R ⁹,=CHCOOR ^5b,=CHPO ₃R ^6a ₂Or=CHCN; Or R ³Or R ⁴In one represent amino and another representation carboxy;

X ' represents key, CH ₂Or CO;

M is the integer of 1-3;

R ⁵, R ^5a, R ^5b, R ^5c, R ⁷, R ⁸And R ⁹Independent is hydrogen atom; Optional (1-6C) alkyl that replaces; Optional (2-6C) thiazolinyl that replaces; Optional (2-6C) alkynyl that replaces; The optional aryl that replaces; The optional heteroaryl that replaces; Non-aromatic carbon ring group; Non-heteroaromatic group; With one or two monocyclic aryl or heteroaryl-condensed non-aromatic monocycle carbon ring group; Or with one or two monocyclic aryl or heteroaryl-condensed non-aromatic monocyclic heterocycles group;

R ⁶And R ^6aIndependent hydrogen or (1-6C) alkyl represented;

R ¹⁰Be hydrogen or fluorine; With

R ¹¹Be hydrogen, fluorine or hydroxyl.

2. according to the compound or the salt of claim 1, condition is that this compound or salt are not CR for X wherein ³R ⁴, R wherein ³Be fluorine, and R ⁴Be hydrogen, p is 1, and Q is the compound of L-alanyl; Or its pharmaceutically useful salt.

3. according to the compound or the salt of claim 1 or 2, wherein

A is H-(Q) _p-;

When occurring at every turn, Q independently is selected from aminoacyl;

P is the integer of 1-3;

X is O, S, SO, SO ₂Or CR ³R ⁴

R ³Be fluorine or hydroxyl, and R ⁴Be hydrogen; Or R ³And R ⁴Representative=O together;

R ¹⁰Be hydrogen or fluorine; And

R ¹¹Be hydrogen, fluorine or hydroxyl.

4. according to each compound or salt among the claim 1-3, wherein Q is the aminoacyl derived from natural amino acid.

5. according to each compound or salt among the claim 1-4, wherein X is SO ₂

6. according to each compound or salt among the claim 1-4, wherein X is CR ³R ⁴, R ³Be fluorine, and R ⁴Be hydrogen.

7. according to each compound or salt among the claim 1-4, wherein X is CR ³R ⁴, R ³Be hydroxyl, and R ⁴Be hydrogen.

8. according to the pharmaceutically useful salt of claim 1, it is the acid-adducting salt that acid makes, and this acid provides pharmaceutically useful negatively charged ion; The alkali adduct that alkali makes, this alkali provides pharmaceutically useful negatively charged ion for the compound that contains acid moieties; Or contain the zwitterionic compound of reverse band electric group.

9. according to the compound of claim 1, wherein

A is H-(Q) _p-;

Q is the L-alanyl;

P is 1;

X is SO ₂Or CR ³R ⁴

R ³Be fluorine and R ⁴Be hydrogen;

R ¹⁰Be hydrogen; And

R ¹¹Be hydrogen;

Or its hydrochloride, tosylate, mesylate, esilate, benzene sulfonate or single sodium salt.

10. according to the pharmaceutically useful salt of claim 9, its be (1R, 4S, 5S, 6S)-4-(the amino propionyl of 2 ' S-) is amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylate hydrochlorate or (1R, 4S, 5S, 6S)-4-(2 ' S-2 '-amino propionyl) is amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylic acid tosylate.

11. according to the compound of claim 1, its be (1R, 4S, 5S, 6S)-4-(2 ' S-4 '-methylthio group-2 '-amino butyryl radicals) is amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylic acid or its pharmaceutically useful salt.

12. according to the compound of claim 11, its be (1R, 4S, 5S, 6S)-4-(2 ' S-4 '-methylthio group-2 '-amino butyryl radicals) is amino-2,2-dioxo-2 λ ⁶-thia-dicyclo [3.1.0] hexane-4,6-dicarboxylic acid monohydrate.

13. according to the pharmaceutically useful salt of claim 1, it is 1S, 2S, 4S, 5R, 6R-2-(the amino propionyl of 2 ' S-) amino-4-hydroxy-dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate.

14. according to the compound of claim 1, wherein

A is H-(Q) _p-;

Q is the L-alanyl;

P is 1;

X is CR ³R ⁴

R ³Be fluorine and R ⁴Be hydrogen;

R ¹⁰Be hydrogen; And

R ¹¹Be hydrogen;

Or its pharmaceutically useful salt.

15. according to the compound or the salt of claim 14, it is selected from:

A) 1R, 2S, 4R, 5R, 6R-2-(2 ' S-2 '-amino propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylate hydrochlorate;

B) 1R, 2S, 4R, 5R, 6R-2-(2 ' S-2 '-amino propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid mesylate;

C) 1R, 2S, 4R, 5R, 6R-2-(2 ' S-2 '-amino propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid esilate;

D) 1R, 2S, 4R, 5R, 6R-2-(2 ' S-2 '-amino propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid benzene sulfonate;

E) 1R, 2S, 4R, 5R, 6R-2-(2 ' S-2 '-amino propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid tosylate;

F) 1R, 2S, 4R, 5R, 6R-2-(2 ' S-2 '-amino propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid; With

G) 1R, 2S, 4R, 5R, 6R-2-(2 ' S-2 '-amino propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid list sodium salt.

16. according to the pharmaceutically useful salt of claim 15, its be (1R, 2S, 4R, 5R, 6R)-2-(2 ' S-2 '-amino propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid mesylate.

17. according to the pharmaceutically useful salt of claim 16, its be (1R, 2S, 4R, 5R, 6R)-2-(2 ' S-2 '-amino propionyl) amino-4-fluorine dicyclo [3.1.0] hexane-2,6-dicarboxylic acid mesylate monohydrate.

18. one kind prepares the formula I compound described in claim 1 or the method for its pharmaceutically useful salt, comprises acidylate formula compound (ii):

Aminoacyl with corresponding formula III carries out acidylate

Pg ^N-A- (III)

Pg wherein ^NBe nitrogen-protecting group group;

Then, for above-mentioned any one step, when using blocking group that functional group is protected, remove blocking group;

Then, for above-mentioned any one step: when needing the pharmaceutically useful salt of formula I compound, make this formula I compound of alkali form and provide the acid of pharmaceutically useful counterion to react; Or, make this compound of sour form and pharmaceutically useful cationic alkali reaction is provided for the formula I compound that contains acid moieties; Or for the zwitterionic compound of formula I, the formula I compound of neutralizing acid adduct form or alkali adduct form; Or adopt any other traditional method.

19. a method that influences the metabotropic glutamate receptor that cAMP connects in the patient body, it comprises the patient who the compound administration of the claim 1 of medicine effective quantity is regulated the excitatory amino acid neurotransmission in needs.

20. the method for the formula II compound of an effective dosage,

Wherein, X and R ¹⁰As the definition in the claim 1,

It comprises the patient who the compound administration of the claim 1 of medicine effective quantity is regulated the excitatory amino acid neurotransmission in needs.

21. a method for the treatment of patient's nervous system disorders, it comprises that compound administration with the claim 1 of medicine effective quantity is in the patient of this treatment of needs.

22. the method for claim 21, wherein said nervous system disorders are the heart bypass operation and transplant the brain defective that causes; Cerebral ischemia; Spinal cord injuries receptor; Head trauma; Presenile dementia; Huntington Chorea; Amyotrophic lateral sclerosis; The dementia that acquired immune deficiency syndrome (AIDS) is brought out; Term anoxic; The hypoglycemia nerve injury; Eyes wound and retinopathy; Cognitive illnesses; Congenital and drug-induced Parkinson's disease; Muscle spasm; Migraine; The urinary incontinence; Drug tolerance; Give up; Drug withdrawal and medicine are thirsted for; Smoking cessation; Vomiting; Cerebral edema; Chronic pain; Insomnia; Faint from fear; The Tourette syndromes; ADHD (Attention Deficit Hyperactivity Disorder) and tardive dyskinesia.

23. being drug tolerance, withdrawal, drug withdrawal and medicine, the method for claim 22, wherein said nervous system disorders thirst for; Or smoking cessation.

24. a method for the treatment of patient's mental disorder, it comprises that compound administration with the claim 1 of medicine effective quantity is in the patient of this treatment of needs.

25. the method for claim 24, wherein said mental disorder are that schizophrenia, anxiety and relative disease, depression, bipolarity are unusual, mental anomaly and compulsive disorder.

26. the method for claim 25, wherein mental disorder is anxiety and relative disease.

27. a pharmaceutical preparation comprises and pharmaceutically useful carrier, thinner or excipient bonded formula I compound or its pharmaceutically useful salt.