TW200400815A - Prodrugs of excitatory amino acids - Google Patents

Abstract

This invention relates to synthetic excitatory amino acid prodrugs and processes for their preparation. The invention further relates to methods of using, and pharmaceutical compositions comprising, the compounds for the treatment of neurological disorders and psychiatric disorders.

Description

Description of the invention: [Technical field to which the invention belongs] The present invention proposes a synthetic stimulating amino acid prodrug (compound of the formula) and a method for preparing the same. The invention further relates to a method of use and a pharmaceutical composition comprising a compound of formula I for the treatment of neurological and psychiatric disorders. [Prior art] Treatment of neurological or psychiatric disorders, such as anxiety, has been linked to the selective activation of metabotropic stimulating amino acid receptors. For example, it has been revealed that (+)-4-amino-2-sulfobicyclo [3.1.〇] hexane-4,6-dicarboxylic acid is an active mGlnR2 receptor agonist, u, S. Pat No. 5,688,826 (the 826 patent), published November 18, 1997. In addition, (+) _ 2_amino · 4-fluorobicyclo [3.1.0] hex; t yuan-2,6-Yifei has been revealed as an active mGluR2 receptor agonist in US Pat. No. 5,958,960 (patent 960), published on September 28, 1999. The present invention proposes a prodrug of the mGluR2 receptor agonist compound, which can enhance the strength of individual parent compounds in the body, and allows the parent compound to have a higher oral exposure. The compounds of the present invention represent the best way to maintain the safety and efficacy of the mGlUR2 receptor agonists disclosed above and to increase the oral bioavailability. Synthetic stimulating amino acid prodrugs and their preparation methods are disclosed in the PCT case Nos. PCT / US 01/45866 and PCT / US 02/00488. [Summary of the Invention] The present invention proposes a compound of formula I 85435 200400815 Η

Wherein A is H- (Q) P-; Q is each independently selected from amino fluorenyl groups; P is an integer from 1 to 10; X is 0, S, so, so2 * cr3r4; R fluorinated, X OR , S03H, Tetra-5-yl 'cn, P03R62, light group or No2, and R4 is hydrogen; or R3 and R4 each represent fluorine; or rule 3 and R4 together represent = 0, = NOR7 or = CR8R9; or y or "of—represents an amine group and the other represents an Ik group 'or R represents N3' (CH2) mCO〇R5a, € Cfj2) mP〇3; R6a2 'NHCONHR ^ > mS〇2R5e' and ... Represents hydrogen; or together with ruler 4 = CHC00R5b, = CHP03R6a24 = CHCN; X 'represents a bond, CH24C0; m is an integer from 1 to 3; R5' R5a 'R5b' R5e 'R7' R8 & r9 is independent Hydrogen atom; (1-6C) alkyl group substituted as required; (2-6C) dilute group substituted as required; (2-6C) alkynyl group substituted as needed; aromatic group substituted as needed; Heteroaromatic groups to be substituted; non-aromatic carbocyclic groups; non-aromatic heterocyclic groups; non-aromatic monocyclic carbocyclic groups' are dense with one or two monocyclic aromatic or heteroaromatic groups Together; or non-aromatic monocyclic heterocyclic group, thick with one or two monocyclic aromatic or heteroaromatic groups 85435 200400815 R6 and R6a independently represent hydrogen or (1-6C) alkyl;

R1Q is hydrogen or fluorine; and R is hydrogen, fluorine or meridian; or a pharmaceutically acceptable salt thereof. The present invention also proposes a compound of formula I, wherein the substituent is as defined above, with the limitation that the compound is not a compound in which X is CR3R4, wherein R3 is fluorine and ... is hydrogen, P is 1 and Q is L-propylaminomethyl; or The pharmaceutically acceptable invention of the present invention also proposes a compound of formula I, wherein A * H_ (Q) p_; (^ is independently selected each time from an aminofluorenyl group; P is an integer from 1 to 3; X is 0, s, so, s〇2 ^ CR3R4; R3 is fluorine or hydroxyl, and R4 is hydrogen; or rule 3 and rule 4 together represent = 0, · hydrogen or fluorine; and R11 is hydrogen, fluorine or hydroxyl; or pharmaceutically acceptable Accepted salts. The present invention also proposes a compound of formula 1, where A is H- (Q) p-; (^ is! ^ Amine test group; P is 1; X is CR3R4; r3 is fluorine and R4 is hydrogen; R1Q Is hydrogen; and Rii is hydrogen; or a pharmaceutically acceptable salt thereof. It should be understood that the 'compounds of formula I contain at least four asymmetric carbon atoms. The invention includes' all isomeric forms' including each enantiomer And its mixtures, the previous drug forms of the compounds disclosed in the patent, such as 1 SR, 4RS, 5RS, 6R: S-4 · ¾t-continylfluorenylbicyclo [3 · 1 · 〇] hexane) -4,6-di Carboxylic acid. ^ 月 进 — A further aspect is to propose a pharmaceutical formulation containing a compound of formula I or a pharmaceutically acceptable salt thereof, plus a pharmaceutically acceptable carrier, diluent or elixir. A further aspect of the present invention is that it can affect Patient CAMP-linked stimulus 85435 200400815 generation "radioactive glutamine red (11 ^ 131) 〇1] * (^ 丨 0§11 ^ 111316) receptor method, this method includes the need for a regulated stimulating amine Patients with basic acid neurotransmission are administered a compound of formula 丨 in a pharmaceutically effective dose. The present invention also proposes the use of a compound of formula [medicine] Yiying # patient eAMp_ linked metabotropic glutamate receptor. A further aspect of the invention is to propose a method for administering an effective dose of a compound of formula 11. This includes administering a pharmaceutically effective dose of a compound of formula J to a patient in need of a regulated stimulating amino acid neurotransmission. The invention also proposes formula 1 It can be used as a medicine to administer an effective dose of a compound of formula II. A further aspect of the present invention is to propose a method for treating patients with neurological disorders. This method includes administering medicine to patients in need of this therapy. An effective dose of a compound of formula 1. The invention also proposes the use of a compound of formula I, which can be made into a medicine for treating patients with diabetic disorders. A further aspect of the invention is to propose a method for treating patients with mental disorders. Patients in need of this treatment are administered a medicinal effective dose of Formula I. The present invention also proposes the use of a compound of formula I, which can be made into drug 6 to treat patients with psychiatric disorders. * Formula 1 is similar to the previous chemical method The structure of the production system in the art is similar to that of white. ♦ The known method of clothing σ, or use the novel method described here. The method for preparing the compound of formula 1 and the intermediates are as defined above, and are invented in 7 steps. JL, 甲, and A are generally defined as the meaning of a group unless otherwise stated. The present invention proposes a method for preparing a compound of formula I. This method includes a compound of formula (ii) 85435 200400815

Diluted with amino group equivalent formula III

PgN-A_ (III) where PgN is a nitrogen-protecting group, and A is as defined above; then, for any of the above steps, when the functional group is protected with a protecting group, the protecting group is removed; after that, for the above Any step; when a pharmaceutically acceptable salt of a compound of formula is needed, react the basic form of the compound of formula I with a pharmaceutically acceptable counter ion that can generate an acid; or for a compound of formula with an acidic moiety By reacting the acidic form of the compound of formula I with a base, a pharmaceutically acceptable cation can be formed; or for a compound of formula Ϊ of a zwitterion, it can neutralize the acid addition salt form or test addition salt form ; Or take advantage of any other traditional steps. The present invention also proposes a compound of formula I, wherein \ is 0 'and R 1 is fluorine, and another variable is as defined above. [Embodiments] The compounds of the present invention have been found to be useful prodrugs of the compounds, which are selective agonists of endogenous glutamate receptors, and thus can be used to treat diseases of the nervous system, such as neurological diseases , Such as neurodegenerative diseases; I as antipsychotics, anxiolytics, drugs _ withdrawal, anti-paeonia ^ guanjue sedatives and antiemetics. 85435-10-200400815 It should be understood that the compound of formula I contains at least four asymmetric carbon atoms, three of which are on the cyclopropane ring, and one is on the α-carbon of the amino group. Therefore, the compounds of the present invention can exist and separate in enantiomerically pure, racemic or diastereoisomeric mixtures. The amino acid moiety preferably has a natural amino acid configuration, that is, compared to the L-configuration of D-glyceraldehyde. The invention includes pharmaceutically acceptable salts of compounds of Formula I. These salts may be present in combination as the acidic or basic part of the molecule, and may be present as acid additions, primary, secondary, secondary or quaternary ammonium, metal or earth metal salts. Generally, i, acid addition salts are prepared by reacting an acid with a compound of formula I. Alternatively, the acid addition salt can be prepared by reacting the penultimate compound (protected intermediate) with a suitable equivalent of acid 'to produce equivalent salts, which are then reacted to produce compounds of formula I or other salts. The preparation of metal test and soil test metal salts is to react the hydroxide type of the desired metal salt with the compound of formula. Some special foot salts can provide certain blends because of their crystalline form. The amorphous amorphous form of the compound may be hygroscopic. The crystalline form of a pharmaceutical compound is sometimes more demanded 'because it can exhibit more beneficial solid state properties. Acids commonly used to form this salt include inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid or phosphoric acid, or organic acids such as organic carboxylic acids such as glycolic acid, maleic acid, hydroxymaleic acid 'fumaric acid, apples Acid, tartaric acid, citric acid, salicylic acid, ortho-acetoxybenzoic acid 'or organic sulfonic acid, 2-hydroxyethanesulfonic acid, toluene-para-sulfonic acid, methane_sulfonic acid, naphthalene_2 _ Sulfonic acid, benzenesulfonic acid or ethanoic acid. In addition to the pharmaceutically acceptable salts, other salts are also included in the present invention 85435-11-which can be used as intermediates for purification of chelating substances, or can be used to prepare other pharmaceutically acceptable acid addition salts' or can be used For identification, characterization or purification. In the present invention, the compound of formula [] includes a solvate thereof. In particular, compounds of formula 1 include their hydrates. Furthermore, this Maoming includes prodrugs of fluorinated compounds, such as those described in International Case No. PCT / JP 99/03984 'PCT / JP 99/00324 and PCT / JP 01/05550. See International Case Nos. WO / 0012464, WO / 9938839 and 0/0200605. For example, the present invention includes 1S, 2R, 5S, 6S_2 · aminotofluoro-4 · ketobicyclo [3 丄 0] hexane-2,6_ diquinic acid; ls, 2R, 4s, 5s, 6mu -6 -Fluoro-4-hydroxybicyclo [no] hexane · 26_dicarboxylic acid; is, 2r, 3r, 5s, 6s · 2-amino-3-fluorobicyclo [3 · 1〇] hexane_2, 6_dicarboxylic acid; & ls, 2R, 3s, 5s, 6s before 2-amino-6-fluoro-3-hydroxybicyclo [3. I.〇] hexane_2,6_dicarboxylic acid medicine. Various physiological functions have been shown to be affected by excessive or inappropriate stimulation of stimulating amino acid delivery. The letter of the formula of the present invention has the ability to orally treat various neurological disorders related to this condition in mammals, including acute neurological disorders, such as brain defects after cardiac circuit surgery and transplantation, stroke, and brain Hemostasis, spinal cord trauma, head trauma, hypoxia during delivery, cardiac arrest, and hypoglycemic neuron damage. The Hamamatsu type has the ability to treat various chronic neurological disorders, such as Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, dementia induced by Ams_, eyes Bruises and retinopathy, cognitive disorders and primary and drug-induced Parkinson's disease. The invention also proposes a method of treating these disorders, which method comprises administering to a patient in need thereof an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof. 85435 -12- 200400815 The 4I compound of the present invention can treat patients, various other neurological disorders related to folic acid of osmic acid, including muscle cramps, convulsions, migraine, urinary incontinence, pain, premenstrual irritability Disorders (pDD), psychiatric disorders (such as schizophrenia) 'drugs for sex, withdrawal, discontinuation and addiction (such as nicotine, opiates, codeine, benzodiazepines and alcohol), anxiety And related alopecia, vomiting, cerebral edema, chronic pain and bradykinesia. Formulas can also be used as antidepressants and analgesics. Therefore, the present invention also proposes a method for treating these disorders, which method comprises administering to a patient in need thereof an effective dose of a compound of formula I or a pharmaceutically acceptable salt thereof. The definitions of the various terms used in the following definitions and examples are shown below. The general terms used here have their general meanings. Affect ' refers to the effect of a compound of formula II on an irritating amino acid receptor, such as an agonist. " Stirring amino acid receptor refers to a receptor that metabolizes glutamic acid, which is a receptor coupled to a cell effector by a GTP-binding protein. "Camp " _ Linked metabotropic glutamate receptors" refers to metabotropic receptors that are associated with inhibition of adenylate cyclase activity. "Nervous disorder" refers to acute and chronic neurodegenerative diseases. Conditions, including brain defects after cardiac circuit surgery and transplantation, brain hemorrhage (such as stroke caused by cardiac arrest), 'spine trauma', head trauma, Alzheimer's disease, Huntington's disease, muscle Atrophic lateral sclerosis, AIDS-induced dementia, hypoxic 'hypoglycemia neuron damage during production, eye damage and retinopathy, cognitive disorders, primary and drug-induced Parkinson's This term also includes other neurological conditions caused by glutamine dysfunction, including muscle spasms, migraines, urinary incontinence, drug tolerance, withdrawal, discontinuation, and addiction (ie 85435 -13- 200400815 quail tablets). Class, diazepam, nicotine, codeine or alcohol), smoking cessation " area vomiting, cerebral edema, chronic pain, sleep disorders' convulsions, T0urettces syndrome, Attention Disorder and Delay Type of exercise cannot. "Psychiatric disorders" refers to two acute and chronic mental conditions, including scabbard anxiety and related disorders (such as panic attacks and stress-related cardiovascular disorders) 'depression, bipolar disorder Disease, psychosis, obsessive-compulsive psychosis, extensive anxiety disorder, acute stress disorder, and panic disorder. The "effective dose" used herein refers to the content or dose of the compound. Once administered to a patient in single or multiple doses, It enables patients under diagnosis or treatment to increase the effect of supply and demand. Effective but effective] can be easily determined by the attending physician, like a skilled artist, using known techniques and observing results under similar conditions. In determining the effective amount or dose of a compound to be administered, the attending physician must consider many factors, including: the type of mammal, its size, age, and general health; the specific disease involved; the severity or extent of the disease; individual Patient response; special compound administered; mode of administration; bioavailability characteristics of administered preparation; The dosage course used; concomitant use; and other related conditions, but not limited thereto. For example, a typical daily dose of the active ingredient may contain from about 5 mg to about 300 mg. The compound may be administered by various routes, including Oral, transanal, transdermal, subcutaneous, intravenous, intramuscular, buccal or intranasal route. In addition, the compound can be administered by continuous infusion. "Patient" as used herein refers to mammals, such as rats, guinea pigs, Big 85435 -14- 200400815 Rat, dog or human. It should be understood that people are better patients. " Treatment " includes its generally acceptable meaning, which includes prohibition, prevention, prevention and mitigation, cessation or reversal of the symptoms produced. Thus, the method of the present invention includes treatment and prophylactic administration. As used herein The general chemical terms have their general meanings. For example, "(1-6C) alkyl" means a straight or branched group. Examples of (1-6C) alkyl include (1-4C) alkyl such as methyl, ethyl, propyl, isopropyl, butyl and isobutyl. " (2-6C) alkenyl " includes (2-4C) alkenyl, such as allyl. " (2-6C) alkynyl includes (2-4C) alkenyl, such as dipropyl. " "(1-6C) alkyl", "2-6C) alkenyl" and "(2-6C) block" if necessary ; Here, it means that more than one substituent may be present, preferably 1 to 3, the substituent is selected from atoms and groups, which when present in the compound of the formula I, does not prevent the compound of the formula I from regulating the metabolism-promoting property Receptor function of glutamic acid. It can exist in (1-6C) alkyl, optionally substituted (2-6C) fluorenyl, or (2-6C) alkynyl, optionally substituted. Examples of the atom and the group include an aromatic group substituted as required, a heteroaromatic group substituted as required, a non-aromatic carbocyclic group, a non-aromatic heterocyclic group, a non-aromatic monocyclic carbocyclic group, One or two monocyclic aromatic or heteroaromatic groups are fused, and non-aromatic monocyclic heterocyclic groups are fused, and one or two monocyclic aromatic or heteroaromatic groups are fused. "Heteroaromatic" includes 1 to 4 aromatic 5-6 membered rings selected from heteroatoms of oxygen, sulfur and nitrogen, And 5-6 member rings containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen are fused to 5-6 member rings or benzene rings containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen Composition of aromatic bicyclic groups. Examples of heteroaromatic groups are: furanyl, 85435 -15-200400815 thiophenyl, No. 57 phenyl group, isosaki σ group, 11 group 11 group, isopylin group , Imidyl, pyrimidinyl, benzofuranyl, benzothiophenyl, benzimidazolyl, benzoyl sigma group, benzopyridyl sigma group and '^ 丨 11 aryl group. "Aromatic group" ; Including phenyl and polycyclic aromatic carbocyclic rings, such as fluorenyl. "Optionally substituted" as in "optionally substituted heteroaromatic" and "optionally substituted aromatic" indicates that more than one substituent may be present, the substituent being selected from Atoms and groups that, when present in a compound of formula I, do not prevent the compound of formula I from regulating the function of the metabotropic glutamate receptor. Examples of atoms and groups that may be present in the optionally substituted heteroaromatic or optionally substituted aromatic groups are: amino, hydroxyl, nitro, halogen, (1-6C) alkyl, ( 1-6C) alkoxy, (1-6C) alkylthio, carboxyl, (1-6C) alkoxycarbonyl, aminoamido, (1-6C) alkylamido, (1-6C) alkyl Sulfonyl, (1-6C) alkyl sulfonylamino, optionally substituted phenyl, phenoxy, phenylthio, phenyl-continuous group 'phenyl-continuous Si-amino' toluene Testylamino ^ (1-6C) fluoro dry group and (1-6C) fluorotrioxyl group. Examples of special values are: amine, triphenyl, fluorine, chlorine, bromine, iodine, methyl, methoxy, methylthio, carboxyl, ethylamino, ethylamino, nitro, ethyl , Phenoxy, phenylthio, phenylsulfonyl, methanesulfonylamino, and trifluorofluorenyl. Examples of the optionally substituted aromatic group include: 1-naphthyl, 2-fluorenyl, phenyl, 2-biphenyl, 3-biphenyl, 4-biphenyl, 2-hydroxybenzene , 3-hydroxyphenyl, 4-hydroxyphenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,4-difluorophenyl, 3,4-difluorophenyl, Pentafluorophenyl, 2-chlorophenyl, 3-chlorophenyl, 4-muryl '2,4-dioxafluorenyl' 3,4-dioxophenyl '3-chloro-4-aminophenyl , 3,5-dichlorophenyl, 2-bromophenyl, 3-bromophenyl, 4-bromophenyl, 2-methylphenyl, 85435 -16- 200400815 3-methylphenyl, 4-methyl Phenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methyllactate · group '2,3- · —methoxybenzyl, 2,5-dimethoxyphenyl, 3,4- Dimethoxyphenyl, 3,5-dimethoxyphenyl, 2-trifluoromethylphenyl, 3-trifluoromethylphenyl '4-trifluoromethylphenyl' 2-fluoro-3-tri Fluoromethylphenyl, 3-trifluoromethyl-4-fluorophenyl, 3-trifluorofluorenyl-5-fluorophenyl '2-fluoro-5-trifluorofluorenylphenyl, 2-phenoxybenzene , 3-phenoxyphenyl, 3-carboxyphenyl and 4-carboxyphenyl. And "non-aromatic carbocyclyl" includes monocyclic groups such as (3-10C) cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclonon Or cyclodecyl, and condensed polycyclic groups, such as 丨 _adamantyl or 2-adamantyl, 1-alkyl, 2- 2-alkyl, 4a-alkyl, and bicyclic [3.3〇 ] Octyl group, -2-yl group or -3-yl group, bicyclic [4_3〇] nonyl group, _2 group, _3 group or _7 group, bicyclic group [5.3.0] decyl -1-yl, _2_yl, _3_yl, _4_yl, _8_yl or _9_yl and bicyclic [3.3.1] nonyl, _2_yl, dongyl or dongyl. " Non-aromatic heterocyclic group "includes a 4- to 7-membered ring containing 丨 or 2 heteroatoms selected from oxygen, sulfur, and nitrogen, such as a nitrogen atom or a _2_yl group, and pyrrolidine_ 丨 _ Radical, _2_yl or -3-yl, hexahydropyridine, 丨 _yl, · 2_yl, _3_yl or aryl, hexahydronitro-1-yl, -2-yl, -3-yl or _ 4-yl, oxo_2_ or _3_, tetrahydrofuran-2-yl or -3-yl, tetrahydropyran_2_yl, _3_yl, or _4-yl, hexahydroxan Fluoren-2-yl, -3-yl or · 4-yl, thio_2_yl or _3_, tetrahydrothiobenzene_2_yl or -3-yl, tetrahydrothiopyranyl, _3 _ Group or _4_ group, hexahydrothiazine_2_ group, -3- group or -4- group, piperidin _ 丨 _ group or _2_ group, morpholine _ 丨 _ group, group or- 3-yl, thiomorpholine _ 丨 _ group, _2_ group or group, tetrahydropyrimidin _ 丨 _ group, -2-yl, 4-yl or _5 group, imidazoline _ 丨 _ group, _2 _Yl or _4_, imidazolidin-1-yl'-2-yl or _4_yl, oxazoline_2_yl, _3._yl, _4_yl or alkynyl, 85435 -17- 200400815 Pyrazolidin-2-yl, -3-yl, -4-yl or -5-yl, oxazoline-2-yl, -3-yl, -4-yl or -5-yl, or pyrazolidine _2_ group, _3_ group, _4_ group or _5_ group. Non-aromatic monocyclic ring breaking Fused with one or two monocyclic aromatic or heteroaromatic groups "includes (3-10C) cycloalkyl fused with a benzene ring or an aromatic 5-6 membered ring containing i to 4 selected from oxygen, sulfur And nitrogen heteroatoms, such as hydroindenyl, 2,3,4-tetrahydrofluoren-1-yl or -2-yl, 5,6,7,8-tetrahydroquinoline 5-yl, -6 _ Group, _7_ group or -8_ group '5,6,7,8-tetrahydroisoquinoline_5 · group, · 6_ group, _7_ group or _8_ group, 4,5,6,7_ Tetrahydrobenzopyrimidin-4-yl, -5-yl, -6-yl or -7-yl, dibenzo [2,3,6,7] cycloheptan-1-yl or -4-yl, Dibenzo [2,3,6,7] cyclohepta-4-ene_ 丨 _ group or -4_ group, or fluorenyl group. "A non-aromatic monocyclic heterocyclyl group fused with one or two monocyclic aromatic or heteroaromatic groups" includes a 4- to 7-membered ring containing one or two heteroatoms selected from oxygen, sulfur, and nitrogen, Fused with a benzene ring or an aromatic 5-6 membered ring containing [to 4 heteroatoms selected from oxygen, sulfur, and nitrogen, such as 2,3-dihydrobenzopyran-2-yl, radical or _4 _ Group, 咄 9 group, 1,2,3,4-tetrahydro α quinine_ι_ group, _2_ group, _3 · group or _4 · group, 9, ι〇_ dihydroacridin-9- group or -10-yl, 2,3_dihydrobenzothiopyran_2_yl, _3_yl or -4-yl, or dibenzothiopyran_4_yl. " Nitrogen-protecting group " as used herein, and "PgN" means a group which can protect or block non-desirable reactions in the nitrogen-antagonizing synthetic step. The choice of nitrogen protection base used depends on the application of the subsequent reaction steps (where protection is needed), and is within the knowledge of the skilled person. Commonly used nitrogen protecting groups are disclosed in T.W. Greene and P.G.M. Wuts, Protective Groups In Organic Synthesis, 3rd Ed. (John Wiley & Sons, New York (1999)). A more preferred nitrogen protecting group is tertiary butoxycarbonyl. 85435 -18- 200400815 "Carboxyl protecting group" as used herein and represented by "PgG" refers to one of the ester derivatives of carboxylic acid groups, which is commonly used to block or protect carboxylic acid groups. At this time, the reaction is in other compounds. On the functional group. Special groups include, for example: methyl, ethyl, tertiary butyl, uryl, methoxymethyl * trimethylphenyl, and the like. Further examples of this group can be found in T.W. Greene and P.G.M. Wuts, Protecting Groups In Organic Synthesis, 3rd Ed. (John Wiley & Sons, New York (1999)). Preferred carboxy-protecting groups are methyl and ethyl. Esters are disintegrated using traditional procedures, which do not affect other parts of the molecule. "Hydroxy protecting group" refers to a group understood by those skilled in organic chemistry in Greene and Wuts Chapter 2. Representative hydroxyl protecting groups include, for example, ether groups, substituted ethyl ether groups, and isopropyl groups. Ether groups, phenyl and substituted phenyl ether groups, benzyl and substituted fluorenyl ether groups, alkylsilyl ether groups, ester protecting groups, etc. The hydroxy protecting group used is not critical, as long as it is derived Under the reaction conditions, the hydroxyl group can be stabilized at other intermediate positions, and can be selectively removed at a suitable point without damaging other parts of the molecule, including any other hydroxyl protecting groups. &Quot; Amine "Amino group" means an amino group derived from an amino acid, and is selected from the group consisting of natural and unnatural amino acids defined herein. Natural amino acids can be neutral, positive or negative, Depending on the substituents on the side chain. &Quot; Neutral amino acid " means an amino acid containing an uncharged side chain substituent. Exemplary neutral amino acids include alanine, valine, and leucine , Isoleucine, proline, phenylalanine, tryptophan, Thiamine, Glycine, Serine, Threonine, Cysteine, Glutamine and Asparagine. "Positive Amino Acid" means amino acid whose side chain substituents are at physiological pH The value is positively charged. Exemplary positive amines 85435 -19- 200400815 Basic acids include lysine, arginine and histidine. "Negative amino acid" means that the side chain substituents of the amino acid are negatively charged at physiological pH. Exemplary negative amino acids include aspartic acid and glutamic acid. A preferred amino acid is an alpha-amino acid. The most preferred amino acid is alpha-amino acid and has an L stereochemistry on the alpha-carbon. Exemplary natural α-amino acids are valine, isoleucine, proline, phenylalanine, tryptophan, methionine, glycine, serine, threonine, cysteine , Glutamic acid, asparagine, glutamine, lysine, lysine, arginine, histamine, aspartic acid, and glutamic acid. "Unnatural amino acid" means amino acid which has no nucleic acid carbon. Examples of non-natural amino acids include the D-isomers of natural α-amino acids as shown above; Aib (aminobutyric acid), pAib (3-aminoisobutyric acid), Nva (n-valine), β-Ala, Aad (2-aminoadipate), PAad (3-aminoadipate), Abu (2-amine Butyric acid), Gaba (γ-amino butyric acid), Acp (6-amino acid), Dbu (2,4-diamino butyric acid), α-amino pimelic acid, TMSA (trimethylsilane) -Ala), alle (be-isoleucine), Nle (n-leucine), tert-Leu, Cit (citrulline), Orn, Dpm (2,2'-diamine pimelic acid) 'Dpr (2,3-diaminopropionic acid)' α- or P-Nal, Cha (cyclohexyl-Ala), hydroxyproline, Sar (sarcosine), 0-methyltyrosine, benzene Glycine and others; cyclic amino acids; Nα-alkylated amino acids where Nα-alkylated amino acids are Na- (l-10C) alkylamino acids, such as MeGly (Na-methyl Glycine), EtGly (Na-ethylglycine) and EtAsn (Na-ethylaspartamine) and amino acids in which the α-carbon carries two side chain substituents. The demonstration is not natural Alpha-amino acids include D-propylamine , D-leucine and phenylglycine. The names of natural and non-natural amino acids and their residues are based on 85435 -20- 200400815 recommended by IUPAC-IUB Joint Commission on Biochemical Nomenclature (JCBN). "Nomenclature and Symbolism for Amino Acids and Peptides (Recommendations, 1983) " European Journal of Biochemistry, 13 8, 9-37 (1984). To the extent that the names and abbreviations used for the amino acids and their residues applied in this case and the attached patent application are different from those shown, the different names and abbreviations will be clearer. Although the compounds of formula I are all useful active mGluR2 receptor agonists, certain compounds are still preferred. The preferred types are specified below. A) Q is glycine, propylamine, valine, leucine, isoleucine, proline, amphetamine, succinyl, tryptamine, methylthio Amidino, lysamino or serine. B) Q is propylamine. C) Q is methionamine. D) p is 1. E) p is 2. F) X is S02. G) X is CR3R4. H) R3 is fluorine and R4 is hydrogen. I) R3 is hydroxyl and R4 is hydrogen. J) R3 and R4 together = 0. K) R1 () is hydrogen. L) R1Q is fluorine. M) Rn is hydrogen. N) Compounds are free. O) The compound is a salt. 85435 -21- 200400815 P) The compound is the hydrochloride. Q) The compound is a mesylate. R) The compound is acetic acid salt. S) The compound is a tosylate. It can be described as an additional preferred compound that can be mixed to show the compound.

Compounds of formula 1 are useful for treating disorders in mammals: and compared to humans. The compounds of the present invention can be prepared in a variety of steps, some of which are described in >. The special order of steps required to produce the compound of formula I depends on: the specific compound, the starting compound, and the substituted part are relatively different: j 1 depends on the origin. Certain substituents are clarified in the following scheme, the team quilt, & ‘, and do not want to limit the teaching of the process in any way. As understood by the artisan, the substituent R15 & RW represents a suitable side chain that forms the desired amine group. If the method is purchased, the necessary starting materials for the following processes can be made by stepping shovel. It is selected from the standard techniques of organic and heterocyclic chemistry, similar to the known and structurally similar compounds. The steps in the formulations and examples include novel steps. Flow 1

(]) The compound of formula 1 is converted into a compound of formula π 85435 -22- 200400815 in ϋΜΑ through an enzyme or a hydrolysis process, as shown in scheme I above. In particular, the compound of formula I in crystalline form can be prepared according to the procedure shown in the following scheme 2. Process 2

The diester-protected peptidium compound of formula (iii) is hydrolyzed with a suitable base, such as lithium hydroxide or sodium hydroxide, in a suitable solvent, such as THF or THF / water, to form a di-acid of formula (iv) Protected peptidyl compound. The compound of formula (iv) can be deprotected with a suitable acid in a suitable solvent. This condition can produce di-acid peptide 85435 -23-200400815. The equivalent acid salt of the hydrazone compound is shown in the formula ㉙,, θ King, ,,, and daily solids. χ, which represents 闳 f 曲 4, 丨 2w qu ion. In the absence of crystalline form, the following crystallization occurs, using T from the solvent, using reagents such as vinegar, attracting cation species can form rebel salt =. Post-bleaching ' zwitterionic compounds can be formed from crystalline salt compounds and appropriate test treatments. For example, a diacid protected peptide compound of formula (iv), when treated with a chlorinated gas in a suitable solvent, can form a pure hydrochloride salt, which is: The amorphous hydrochloride compound can be recrystallized from acetone and water to form a crystalline hydrochloride compound. In the case of a crystalline solid, it can be formed directly, and the filtration of the reaction > m compound can form a crystalline salt. Zwitterionic compounds can be formed by treating crystalline hydrochloride compounds with sodium hydroxide; in addition, treatment with sodium formate compounds or tosylate compounds with sodium hydroxide can also generate zwitterionic compounds. The skilled artisan will appreciate that compounds of formula can be prepared in a step where the intermediates shown therein are not isolated. Process 3

85435

H

The diester of formula (ii) is acidified with a compound of formula III with a suitable coupling agent to produce a diester-protected peptidyl compound of formula (iii). Alternatively, this transformation can be achieved using the fluorenyl chloride of the compound of formula -24-200400815 III. Suitable peptide coupling reagents include dicyclohexylcarbodiimide (DCC), 1- (3-dimethylaminopropyl) _3-ethylcarbodiimide (EDC), isobutyl chloroformate'chlorophosphate Diphenyl ester, 2-chloro-4,6-dimethoxy-1,3,5-tricotine (CDMT), bis (2-keto-3-oxazolidinyl) phosphinium chloride and benzo Triazole-1 -yloxytris (dimethylamino) hexafluoromarate scale. Flow 4

In Scheme 4 above, the compound of formula π, a di-acid, is treated with a suitable carboxyl-protecting agent, such as catalytic hydrochloric acid or thionyl chloride and methanol or ethanol, to generate the equivalent di- vinegar. In addition, a compound of formula II is first treated with a nitrogen-protecting agent such as BOC20 'to form a nitrogen-protected compound of formula IX. Next, the compound of formula ⑴ can be treated with a carboxyl-protecting agent, such as methyl iodide, and has been proven 85435 • 25- 200400815, such as potassium carbonate, followed by a nitrogen deprotecting agent, such as hydrochloric acid or trifluoroacetic acid, Compounds of formula (ii) can be produced. In addition, the skilled artisan can confirm that, depending on X, a suitable protective agent is necessary. For example, if X represents CR3R4 ', R3 represents hydroxy, and R4 represents hydrogen, then the farmer can understand that in step 4 of any step i in the process, suitable protecting groups are necessary. Compounds of formula (I) are known in the art. For example, the preparation of these compounds can be described in US Pat. Nos. 5,688,826 (-826 ^^) ^ 5,958,960 (-960 ^ in the previously disclosed method 'for the improvement of the compounds of formula π. This improvement involves the oxidation of sulfur and alcohol, The synthetic route has been optically resolved for each object, and the first improvement as described below is related to the oxidation of the conversion compounds described in column 333 (formula V).

Column 8, rows 22-34 and column 7, which relates to '826 Patent Formula VII

826 patent formula (VII) can be formed, 826 patent formula V

Formula (V) 85435-26-200400815 of the 826 patent has found that the sulfur trioxide-toxidine complex or trifluoroacetic anhydride combined with dmso is preferred for many oxidation methods known in the art. Second, the analysis of compounds of formula III in the '826 patent

Among them, R2 represents a carboxyl group. In column 8, rows 3-7 'and column 6, starting from the first line (Formula III), it was found that (R) -a-methylfluorenylamine and quinine are preferred. . Especially preferred is (R) -a-methylamidamine. Furthermore, it was found that when the sulfide of the compound of formula III in the '826 patent, where X is sulfur, is formed, the compound of formula III in the' 826 patent, where X is sulfonyl, as in column 8 of the '826 patent, It is better to use a basic aqueous system and a combination of hydrogen peroxide and a catalyst. The following examples further illustrate the compounds of the present invention and their synthetic methods. The examples are not intended to limit the invention in any way, and should not be so limited. All experiments were performed under positive pressure of anhydrous nitrogen or argon. All solvents and reagents were purchased from commercial sources and used as such unless otherwise indicated. Anhydrous tetrahydrofuran (THF) can be obtained by distillation from sodium or sodium benzophenone before use. Proton nuclear magnetic resonance (NMR) spectra can be obtained at Bruker Avance II bay-500 at 500 MHz, Bruker Avance I bay-200 at 200 MHz, or Varian Inova / Varian 300 / Varian 400 at 500 MHz. Electron spray mass spectrometry (ESI) was performed on an Agilent MSD / B instrument using acetonitrile / ammonium acetate as the mobile phase. Free atomic impactor 85435 -27- 200400815 spectroscopy (FABMS) was performed on a VVGAB-2SE instrument. Magnetic field analytical mass spectrometry (FDMS) was performed using a VG 70SE or Varian MAT 731 instrument. Optical rotation was measured on a Perkin-Elmer 241 polarimeter. Upstream chromatography on Waters Prep 500 LC is usually performed using a linear gradient of solvent as shown in the text. Tracking of complete or incomplete reactions was performed using thin layer chromatography (TLC). Thin layer chromatography was performed using an E. Merck Kieselgel 60 F254 plate, 5 cm x 10 cm, 0.25 mm thickness. The detection of the point is a combination of UV and chemical detection (disc immersion in ammonium molybdate solution [75 g of ammonium molybdate and 4 g of sparse acid # (IV) in 500 ml of 10% sulfuric acid aqueous solution]) Plate heating). Flash chromatography was performed as described by Still, et al. Still, Kahn, and Mitra, J. Org. Chem. 43, 2923 (1978). For carbon, hydrogen, and nitrogen elemental analysis was determined on the Control Equipment Corporation 440 Elemental Analyzer, or performed by the Universidad Complutense Analytical Centre (Facultad de Farmacia, Madrid, Spain). Melting points were determined on an open glass capillary on a Gallenkamp hot air bath melting point device or on a Buchi melting point device and were not corrected. The abbreviations, symbols and terms used in the examples have the following definitions.

Ac = Ethyl Anal · = Elemental Analysis ATR = Attenuated Total Internal Reflection Bn or Bzl = Preyl Bu = Butyl BOC = Third Butoxycarbonyl calcd = Estimated d2o = Deuterium Oxide 85435 -28- 200400815 DCO Bicyclic Hexylcarbodiimide DCM = 1, 2-dichloromethane DIBAL-H = diisobutylaluminum hydride DMAP = 4-amidoaminopyridine DMF = metamethoxamine DMS0 = dioxosulfoxide DSC = differential scan Calorimeter EDC = N-ethyl-NW-dimethylaminopropylcarbodiimide hydrochloride ES = electron spray Et = ethyl EtOH = ethanol FAB = fast atomic impact (mass spectrometry) FDMS = magnetic field analytical mass spectrometry FTIR = Fourier (Fourier) transformed infrared spectrum H0At = l-hydroxy-7-azepinetriazole H0Bt = l-hydroxybenzotriazole HPLC = high performance liquid chromatography HRMS = high resolution mass spectrometry i-PrOH = Isopropanol IR = infrared spectrum L = liter

Me = methyl

MeOH = methanol MPLC = medium pressure liquid chromatography 85435 -29 200400815 Μρ = melting point ΜΒΒΕ = third butyl methyl ether NBS = N-bromosuccinimide NMR = NMR magnetic resonance PC-TLC two preparative centrifugal thin layer chromatography Ph = Phenyl p. 〇. = I-Pr = isopropyl

Rochelle's salt = Jonathan tartrate rt = room temperature SM = starting material TBS = tertiary butyldimethylsilyl group T E A = diethyl ether

Temp. = Temperature TFA: trifluoroacetic acid THF = tetramethylfuran TLC = thin layer chromatography t-BOC = tertiary butoxycarbonyl

General procedure A EDC coupling between an amine and an N-BOC-aV amine. The starting amino dialkyl ester (compound of formula ii, scheme 3) (1.0 equivalent) was suspended in anhydrous dichloromethane under nitrogen. Then add equivalent N-Boc- (L) -amino acid (1.5-2.0 equivalents), EDC (1.5-2.0 equivalents), HOBt (1.5-2.0 equivalents), and dimethylamino 17 ratio (DMAP, 0.1 -0.2 equivalent). The reaction mixture is at room temperature 85435 -30- 200400815 unless otherwise indicated. The reaction was stirred until it was judged complete by TLC. The mixture was diluted with ethyl acetate, and then washed sequentially with a saturated aqueous solution of NaHC03 and / or an aqueous solution of NaHSCU and brine. Dry over sodium sulfate and evaporate under vacuum. Purify the crude residue (compound of formula m) by silica gel chromatography using a suitable eluent (usually ethyl acetate / hexane).

General procedure R Amine and N-BOC- (L) -Amine-Isobutyrate > Human equivalent N-Boc- (L) -amino acid (1.5 equivalents) in anhydrous dichloroformamidine (1 〇 Ml) of the solution 'at -20 ° C under nitrogen, add N-methylmorpholine (NMM, 15 ΐ, in 1 liter CI ^ Cl2), followed by dropwise addition of isoformic acid (IBCF) , 1.5 equivalents in 5 ml of CHsCl2) was added at a rate such that the internal temperature of the reaction did not exceed -15 ° C. The resulting reaction mixture was at _20. (: Stir for 30 minutes, and then add (1S, 2S, 4S, 5R, 6R) -2- (2, -amino-propanamido) -4-meryl-bicyclo at -20 ° C [ 3.1.0] A solution of hexane-2,6-dicarboxylic acid diethyl ester hydrochloride (10 when f) in Diqijiakeng (10 ml), added at a rate such that the internal temperature of the reaction does not exceed- 1 5 ° C. Once completely added, 'remove the cooling bath and judge the completeness of the reaction mixture by TLC at room temperature. Dilute the reaction mixture with ethyl acetate' and wash sequentially with saturated aqueous NaHC03, NaHS04 and brine. After drying over sodium sulfate and evaporation under vacuum, the crude residue is purified by silica gel chromatography using a suitable eluent (usually hexane / ethyl acetate).

General Step C: Remove the equivalent N-Boc diester peptide derivative (compound of formula iii, Scheme 2) (1.0 when 85435 -31- 200400815 amount) of N-Boc and N-Boc. > 1 ^ 011 (10-20 equivalents) in a 1: 1 mixture at room temperature for 4 hours at this temperature. The reaction was diluted with H20 and washed with ethyl acetate. Discard the organic layer. The aqueous phase was adjusted to pH 2 with 1 N HC1 (NaCl was added to the aqueous phase to enhance the extractability if necessary), and the N-boc dicarboxylic acid product (compound of formula iv) was further extracted using ethyl acetate. All organic layers were mixed, washed with brine, dried over MgS04, and concentrated to dryness under vacuum to produce the desired carboxylate product 'as a foamy solid. Dissolve in ethyl acetate and cool at 0 ° C. The reaction mixture was filled with anhydrous HC1 gas until saturated with HC1. The resulting reaction mixture was stirred at 0 ° C. for 4 hours. The fully deprotected peptide derivative (compound of formula I) was separated as N1 under filtration like a hydrochloride or the reaction mixture was concentrated to dryness, titrated with ethyl acetate or Et20, and concentrated to a white powder. If necessary, in order to remove the residual solvent and excess HC1 'product, it can be recombined in HiO, frozen, and then lyophilized to produce the desired hydrochloride product. Preparation of fluorene (1R, 4S, 5 $ "S) -4-amino-2,2-diketone -difluorene" 31〇 · | hexane didiethyl ester

f ^ COjCHjCH.

NK p- (lR, 4S, 5S, 6S) -4-amino-2,2-diketo_2λ6-4 _bicyclo [3.1.0] hexane-4,6-dicarboxylic acid (10 g '42.5 millimoles, U.S. Pat. No. 5,688,826) to 100 ml of a slurry of 2 Β ethanol, dropwise added sulfite 85435 -32, 200400815 醯 dichloride (15.5 Ml, 212.6 mmol) and rinse with 40 ml ethanol. The slurry was heated to reflux and stirred overnight. Analysis of the concentrate by 500 MHZ lH NMR (CDsOD) revealed that the starting material and intermediate monoester were completely consumed. The resulting solution was allowed to cool to room temperature and then concentrated to a gelatinous residue. EtoAC (50 ml) was added to gelatin, and further concentrated to a solid, which was then diluted with another 94 ml of Et0AC. A 15% sodium carbonate aqueous solution (; 70 ml) was slowly added to the mixture by hand's shaking to gradually generate the dissolution, so that the final pH reached 7.95. Before the layers were extracted, the resulting sodium carbonate precipitate was filtered. The aqueous layer was extracted with EtoAc (2 x 100 mL). The combined organic extracts were washed with brine (1 x 100 mL), dried (MgS04), filtered and concentrated under vacuum to give a pale yellow oil 'which solidified to give the title compound as a white solid. (11.71 g, 95% yield). Recrystallization A mixture of the title compound (200 mg) in EtOAc (800 microliters) was heated to 56 ° C and the dissolution occurred. After stirring at 56 ° C for 15 minutes, heptane (1 ml) was added dropwise to the solution. Remove heat source. Allow the solution to cool to 52 ° C, at which point precipitation occurred. Once cooled, further dilution with heptane (600 microliters) can form a slurry. The resulting slurry was stirred at room temperature for 1 hour and then filtered, washed with heptane (2x500 microliters), and dried overnight at 45 t to yield 145 mg (73% recovery) of the title compound as a white solid. mp 80-83. (: [A] 25D -57.7 ° (c 1.04, CH3OH) 500 MHz NMR (CD3C13) 64.31 (q, 2H, J = 7.0 Hz), 4.20 (m, 2H), 3.78 (d, 1H, J = 15.0 Hz), 3.36 (dd, 1H, J-4.0, 7.0 Hz), 2-93 (dd, 1H, J = 4.0, 7.0 Hz), 2.81 (d, 1H, J = 15.0 Hz), 2.46 (t, 85435 • 33- 200400815 1H, J = 4.0), 1.34 (t, 3H, J = 7.0), 1.30 (t, 3H, J = 7.0). 13C NMR (125 MHz, CD3C13) 6171.68, 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), 851.25 (s) cm-丨. Analysis and estimate (: 11111 small 068: € :, 45.35; 11, 5.88; team 4.8 measured values: (:, 45.02; H, 5.75; Ν, 4 · 82 ° Preparation 2 (1 ^ 4,3 ,,, 1,58,68) -4- (23-Third-butanil amine propyl propylamine amine) -2,2-di.keto-2λ6 -»Sam-Second Ring「 3.1.01 烧烧 -4,6- 二 # 醢 二 λ 西 ^ V / 0

H3CH2C〇2C

氺: 601 ^ alanine (43.52 g, 230 mol) and ^ -Methylmorpholine (25.5 ml, 232 mol) in 457 ml of dichloromethane at -30 ° C under gas Isobutyl chloroformate (30.4 ml, 234 mmol) was added dropwise to the solution over 10 minutes. The resulting thin slurry was stirred at -25 to -30 ° C for 30 minutes, at which time it was added at 25 minutes. (1 foot, 43,58,63) -4-Amino-2,2-diketo-2 human 6-11 plug_bicyclo [3.1.0] has arrived at -4,6-diacetic acid diethyl Ester (63.90 g, 219 mmol, Preparation 1) in 213 ml of dichloromethane, so that the reaction temperature does not exceed _25 85435 -34- 200400815 C. Once the% is fully added ', the cooling bath is removed and allowed to stir at ambient temperature for 60 minutes. At this time, the reaction temperature reaches 19t and the color becomes light orange. The reaction was treated with 350 ¾ liters of 1 N HC1 and the layers were separated. The organic layer was washed with saturated NaHC03 (1 X 350 ¾ liters) and brine (1 x 350 ml), dried (Na2S〇4), filtered, and then reduced under vacuum to a white foam (1.2 g, 104%). Ή NMR (300 MHz, CDC13) 6: 7.62 (brs, 1H), 4.90 (brd, 1H, J = 7.1 Hz), 4.34-4.10 (m, 6H), 3.39 (ddd, 1H, J = 7.2, 3.9, 1.0 Hz), 3.00 (dd, 1H, J = 7.1, 3.9 Hz), 2.90 (brd, 1H, J = 14.9 Hz), 2.43 (t, 1H, J = 4.1 Hz), 1.46 (s, 9H), 1.31 (m, 9H) »13C Bandit (75 MHz, CDC13) 5: 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 3 (111, 48, ^ 8) -4- (218-111th amine amine-and fluorenylamino) -2.2-diamidino-yl-2X6-g plug-bicyclo 3 · 1 · 〇 1 p, Burning4.6- Ermiao St

(lR, 4S, 5S, 6S) -4- (2'S-tertiary butoxycarbonylamino-propanamido) _2,2_diketo-2λ6-fluorene-bicyclo [3.1.0] hexane_4 A solution of diethyl 6-dicarboxylate (1814 g, 392 mmol, theoretical 2) in a solution of 292 ml of THF at room temperature, and 490 ml (980 mmol) of sodium hydroxide were added. The two-phase mixture 85435 -35 · 200400815 was stirred vigorously at room temperature for 1.25 hours, at which time the reaction was homogeneous. The mixture was diluted with 490¾ liters of ethyl acetate 'and the layers were separated. The aqueous phase was diluted with 49.0 ml of ethyl acetate, and the pH of the mixture was reduced to 1.5 with concentrated HC1. The layers were separated and the aqueous layer was re-extracted with 245¾ liters of ethyl acetate. The combined organic layers were dried (N & δ04), filtered 'and concentrated to give 167.9 g (105%) of the title compound as a white foam. This substance can be used without identification of Examples 1 and 2. Preparation 4 diketone _2λ6 _ ^ _ bicyclo "31〇1 hexane dii6-di # acid dimethyl h3co2c

Thiosulfan digas (6.2 ml, 85.0 mmol) was added dropwise to (1R, 4S, 5S, 6S) 4-amino-2,2-diketo_2λ6-pyrimidine · bicyclo [3.1.0 ] Hexane-4,6-dicarboxylic acid (10.0 g, 42.5 Emole 'Us Pat No. 5,688,826) at Me0, ie 70 = liter, 5. 〇 Stir the suspension quickly. Once completely added, the reaction mixture was slowly warmed to room temperature, and then heated under reflux for 48 hours. The volatiles were removed under reduced pressure, and the residue was partitioned between a saturated solution of NaHC03 (200 ml) and ethyl acetate (400 strokes). The layers were separated and the aqueous layer was extracted with ethyl acetate (= X4 = ml each time). The combined organic layers were dried over κχ 03 and concentrated under reduced pressure to yield 8.10 g (30.8 mmol) of the title compound in a 72% yield. [«] 〇 = -840 (c = 0.5, MeOH) Analytical juice C9H13N06S: c, 41.06; H, 4.98; N, 5.32. Measured value: c, 85435 • 36- 200400815 40.94; Η, 4.93; N, 5.30. MS (ES) m / z 264.0 [M + H] +. 1 ^ 5 [lR, 4S, 5S, 6S) ,,: _ j: (H: carbonyl, amine, _3, _benzyl-propylamido) -2,2-diketobicyclo | ~ 3 Ι.〇ι dimethyl-4,6_diacetate

Prepared according to General Procedure A using commercial N-BOC- (L) -phenylalanine and (111,48,58,68) -4-amino-2,2-diketo-2 human 6-, -Bicyclo [3.1.0] hexane-4,6-dicarboxylic acid dimethyl ester (Preparation 4). The reaction mixture was refluxed overnight. Purified with pc_TLC, 4 mm SiO 2 rotor (10% ethyl acetate / hexane to 100% ethyl acetate), and 0.85 g (88%, 1.67 mmol) of white foam was produced. [《= -35.2o (c = 0.45, CHC13).

lB NMR (300 MHz, CDC13) δ 1.43 (9H, s), 2.38-2.40 (ΐΗ, m) 2.86 (1H, d, J = 15.0 Hz), 2.91 (1H, dd, J = 4.4, 7.3 Hz), 3 〇4 (2H, d, J = 7.3 Hz), 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 = 6.6 Hz), 6.96 (1H, bs), 7.22-7.36 (5H, m). Analysis estimates C23H3ON209S * 0.1 H20: C, 53.92;., 5.94; N, 5 47. I measured: C, 53.62;;, 5.90; N, 5.28. MS (ES) m / z 509 · 16 [M-Η], 411.2 [M_Boc] +. 8543537-200400815

Mj ^ 6 tertiary carbonyl amine even -3, s-methyl-pentyl succinimidine-bicyclo Hoi hexane _4 6_ diboronic acid

Prepared according to general procedure A, using commercially available n_BOC_ (L) _isoleucine and (lR'4S, 5S, 6S) -4-amino-2,2-difluorenyl-2_26-disc-bicyclo [3.1.0] Hexane-4,6-chitosan-methyl acetate (Preparation 4). The reaction mixture was refluxed overnight. Purification with a pc_TLC '4 mm SiO 2 rotor (10% ethyl acetate / hexane to 100% ethyl acetate) yielded 0.75 g (83%' 1.57 mmol) of a white foam. [《= -32_65o (c = 0.49, CHC13). ! H NMR (300 MHz, CDC13) δ 0.91 (3H, t, J = 7.3 Hz), 0.93 (3H, d, J-6.6 Hz), 1.10-1.18 (1H, m), 1.49 (9H, s), 1.42-1.52 (1H, m), 1.81-1.86 (1H, bm), 2.51 (1H, t, J = 4.0 Hz), 2.95 (1H, d, J = 15.0 Hz), 3.06 (1H, dd, J = 4.4, 7.3 Hz), 3.43 (3H, dd, J = 3.7, 7.0 Hz), 3.78 (3H, s), 3.85 (3H, s), 3.82-3.90 (1H, m), 4.2〇 (1H, d, J = 14.7 Hz), 4.94 (1H, d, J = 8.4 Hz), 7.19 (1H, bs). Analysis estimates C20H32N209S: C, 50.41; H, 6.77; N, 5.88. Found: c, 50.32; H, 6.92; N, 5.76. MS (ES) m / z 475.1 [M-Η]. Preparation 7 85435 -38- 200400815 Butoxycarbonyl a certain -3, methyl a-butamidine ^^^^^ 1 ^^ 1 ^^ "3.1.01 hexamethylene.

Prepared according to general step A, using commercially available N_Boc_ (L) _valinic acid and (111,43,5 3,68) -4-amino-2,2-difluorenyl group 2 into 6 " Cyclo-bicyclo [3.1.0] hexane-4,6-dicarboxynomethyl acetate (Preparation 4). The reaction mixture was refluxed overnight. Purified with ρ [_ TLC, 4 mm SiO 2 rotor (10% ethyl acetate) Ester / hexane to 1000 / ethyl acetate), 0.41 g (47 ° / 0, 0.89 mmol) of white foam was formed. [Α] blue = -35.36o (c = 0.51, CHC13). lH NMR (300 MHz, CDC13) δ 0.93 (3H, d, J = 7.0 Hz), 0.96 (3H, d, J = 6.6 Hz), 1.46 (9H, s), 2.06-2.13 (1H, m), 2.50 (1H, t, J = 4.0 Hz), 2.94 (1H, d, J = 15.0 Hz), 3.04 (1H, dd, J = 4.4, 7.3 Hz), 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 = 15.〇Hz), 4.94 (1H, d, J = 8.1 Hz), 7.15 (1H, bs). Analyzed and estimated C19H3 () N2O9S: C, 49.34; H, 6.54; N, 6.06. Found: c, 49.33; H, 6.44; N, 6.05. MS (ES) m / z 461.2 [M-Η] ·. Preparation 8 £ 1. Feet, 48,58,68) -4-(__ 2_5-brother _: = _ Ding Feng% Curcumin _4 | _methyl- 戍 _ | face:

S5435 -39- 200400815 group > 2,2-dione υλ6-4-bicyclo Γ3.1.01 Hexane-4,6-diphosphonic acid dimethyl ^

Prepared according to general step A, using commercially available N-BOC- (L) -leucine and (1-foot, 43,53,63) -4-amino-2,2-di-yl-2 into 6- Edge-Bicyclo [3.1.0] Hexane-4,6-dicarboxylic acid dimethyl ester (Preparation 4). The reaction mixture was refluxed overnight. Purification with pc_TLC, 4 mm SiO 2 rotor (10% ethyl acetate / hexane to 100% ethyl acetate) 'can produce 0.85 g (94%, 1.78 mmol) of white foam. [Tons 3 = -46.15. (. = 1.〇4, CHC13). NMR (300 MHz, CDC13) δ 0.92 (3Η, d, J = 6.2 Hz), 0.95 (3H, d, J = 6.6 Hz), 1.47 (9H, s), 1.42-1.47 (1H, m), 1.63- 1.67 (1H, m), 2.46 (1H, t, J = 3.7 Hz), 2.87 (1H, d, J = 15.0 Hz), 3.04 (1H, dd, J = 4.4, 7.3 Hz), 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 = 15.0 Hz), 4.75 (1H, d, J = 6.6 Hz), 7.43 (1H, bs). Analytical estimates €: 201,132, 093: C, 50.41; H, 6.77; N, 5.88. Found: c, 50.30; H, 6.82; N, 5.75. MS (ES) m / z 475.2 [M-ΗΓ. Preparation 9 Bis-tertiary butoxycarbonyl amine and even hexamidine amine 2, 2-dihydrazone homo- 2λ6-ρ vegetable-Yi soil "3.1.〇1 hexa-fe- 4.6-secondary education--Jiajiu 85435 -40- 200400815

Prepared according to general step A, using commercial N-BOC-Lys (BOC) -OH and (1 foot, 48,58,63) -4-amino-2,2-diketo-2 human 6-pyrimidine -Bicyclo [3.1.0] hexane-4,6-dicarboxylic acid dimethyl ester (Preparation 4). The reaction mixture was refluxed overnight. Purified with PC-TLC, 4 mm SiO 2 rotor (10% ethyl acetate / hexane to 100% ethyl acetate). Produces 1.04 g (93%, 1.76 mmol) of white foam. [c ^ 3 = -32.0o (c = 0.5, CHC13). ! H NMR (300 MHz, CDC13) δ 1.44 (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-4.4 Hz), 2.98-3.20 (4H, m), 3.42 (1H, dd, J = 3.7, 7.0

Hz), 3.76 (3H, s), 3.86 (3H, s), 4.01 (1H, dd, J = 7.7, 13.2 Hz), 4.09-4.19 (1H, m), 4.71 (1H, t, J = 7.3 Hz ), 5.13 (1H, bs), 7.59 (1H, bs) ° Analyzed and estimated CmHwNsOhS: C, 50.75; H, 6.98; N, 7.10. Found: C, 50.36; H, 6.99; N, 6.87. MS (ES) m / z 590.2. Preparation of 10 LLg ^ js, 5s, 6S) -4- "2, s-Third-butoxycarbonylamino group 4'_f A diketone-2λ6-4-diproduct "3 丄 〇11 said 85435 -41--4.6-diboronic acid dimethylamine

Ph was prepared according to the general procedure A, and commercialized n-BOC- (L) -glutamine 200400815 (Trt) -OH and (lR, 4S, 5S, 6S) -4-amino-2,2-di Keto-2λ6-pyrimidine-bicyclo [3.1.0] hexane-4,6-dicarboxylic acid dimethyl ester (Preparation 4). The reaction mixture was refluxed overnight. Purification with a PC-TLC '4 mm SiO 2 rotor (10% ethyl acetate / hexane to 100% ethyl acetate) yielded 0.53 g (48%, 0.72 mmol) of a white foam. [a] 2D3 = -8.0 ° (c = 0.50, MeOH). * H NMR (300 MHz, CDC13) δ 1.42 (9H, s), 1.83-1.88 (1H, m) 5 2.03-2.18 (1H, m), 2.16 (1H, t, J = 4.0 Hz), 2.57-2.64 (1H, m), 2.60 (1H, d, J = 15.0 Hz), 2.64-2.80 (1H, m), 2.88 (1H, dd, J = 4.4, 7.3 Hz), 3.26 (1H, dd ,, J = 4.0, 7.0 Hz), 3.47 (3H, s), 3.76-3.90 (1H, m), 3.81 (3H, s), 4.05 (1H, d, J = 15.0 Hz), 5.47 (1H, bs), 7.02 ( 1H, bs), 7.20-7.35 (15H, m), 8.68 (1H, bs). Analysis estimates C38H43N301 () S: C, 62 · 20; H, 5.91; N, 5.73. Found: c, 61.83; H, 6.09; N, 5.57 ° MS (ES) m / z 731,9 [M-H]-. HRMS estimate C38H43N3n) S [M + Na] +, 756.2567. Measured value: 756.2585 = 85435 -42- 200400815 Preparation 11 (1 foot, 48,58,68, -4- "[1'-Third butyl sulfonium-pyrrolidine-2 | 5" ^^^ · Group 1-2 2,2-diketone _2 nine 6-4-bicyclo ^ 丄 01 hexane-4,6-dicarboxylate ^-^-^ ·

Prepared according to general procedure A, using commercial Boc- (L) -proline (0.61 g, 2.9 mmol) and (lR, 4S, 5S, 6S) -4-amino-2,2- Diketo-2λ6-, said-bicyclo [3.1.0] hexane-4,6-dicarboxylic acid dimethyl ester (0.5 g, 1.9 mmol, Preparation 4). Purification with PC-TLC (ethyl acetate / hexane) gave 0.87 g (99.2%) of the title compound. [〇61.20 (c = 0.49, CHC13). ] H NMR (300 MHz, CDC13) δ 1.51 (9H, s), 1.75-1.98 (3H, m), 2.35-2.5 (2H, m), 2.84 (1H, d, J = 14.7 Hz), 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-H]-. Preparation of 12 (lR, 4S, 5S, 6S) -4- (gU tributyl-oxocarbonylamino-4'_methylsulfan-butylamidoamino) -2,2-diketo-plex 6 Sigh- ~ ^ ring `` 3.1.〇1 has been swept, -4,6-di # acid dimethyl

II 85435 -43- 200400815

BocHN CH3 was prepared according to the general procedure A, using commercially available Boc- (L) -methionine (0.71 g '2.9¾ Mol) and (1R, 4S, 5s, 6S) _4_amino group_2,2_ Diketo_2λ6_tI plug-one ring [3.1.0] hexane-4,6-dimetanoate (o5 g, 1.9 mmol, Preparation 4). Purification using PC-TLC (ethyl acetate / hexane) gave 0.85 g (90.5%) of the title compound. [Chen = -20.4 (c = 0_49, CHC13) NMR (300 MHz, CDC13) δ 1.46 (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 = 4.0 Hz), 2.58 (1H, bs), 2.92 (1H, d, J = 14.7 Hz), 3.01 (1H, dd, J = 4.4, 7.0 Hz, 3.42 (1H, dd, J = 3.7, 7.3 Hz), 3.78 (3H, s), 3.87 (3H, s ), 4.22-4.24 (2H, m), 5.06 (1H, d, J = 7.7 Hz), 7.27 (1H, s). MS (ES) m / z 493.1 [M-Η]. Preparation 13 Tributoxy l-Amine, a third butoxymum-1, 1Η-Θ, blae: 3-violet) -propane .. Amine, a 1.2, 2-dione and its -u6-diethion "3_, 1 〇 丄Hexane-4,6-di # acid dimethyl beep 85435 -44- 200400815

Prepared according to general step A, using commercially available Boc- (L ·) -tryptophan (Boc) (1.1 g, 2.8 mmol) and (lR, 4S, 5S, 6S) -4-amino-2, 2-Diketo-2λ6-pyrimidine-bicyclo [3.1.0] hexane-4,6-dicarboxylic acid dimethyl ester (0.5 g, 1.9 mmol, Preparation 4). Purification with PC-TLC (ethyl acetate / hexane) gave 0-7 g (56.7%) of the title compound. lU NMR (300 MHz, CDC13) δ 1.44 (9Η, s), 1.67 (9Η, s), 2.37 (1H, bs), 2.86 (1H, d, J = 15.0 Hz), 2.88 (1H, t, J = 4.4 Hz), 3.15 (2H, d, J = 6.6 Hz), 3.39 (1H, dd, J = 3.7, 7.0 Hz), 3.73 (3H, s), 3.83 (3H, s), 4.18 (1H, d, J = 14.7 Hz), 4.37-4.44 (1H, m), 5.01 (1H, bd, J = 8.1 Hz), 7.11 (1H, bs), 7.25-7.59 (4H, m), 8.14 (1H, bd, J = 8.4 Hz). [«9.6 (c = 0.51, CHC13). Analysis estimates C3OH39N3OuS * 1.0 C4H802: C, 55.35; H, 6.42; N, 5.70. Found: C, 54.98; H, 6.09; N, 6.07. HRNIS estimates 50 C39H39lSi; 30N NaS, 672.2203. Found: 672 2180. 〇 ^ 1_§ ,, 5 3'68) -4 ,,-12,8: tertiary ^ carbonyl amine and even _3, _ (4_4tributoxymethylenephenyl) -propionyl 1- 2 Shidionone _2λ6_4 _bicyclo 『3 丄 〇1hexane-45-85435 lean-4,6-di # acid dimethyl

200400815 Prepared in the same manner as in step A, using the commercialized 2S-third butoxy oxoylamino group (the first butoxy syloxy-phenyl group) _propionic acid (1.1 g, 29 mmol) and (1 '' 43'58,63) -4-Amino-2,2-difluorenyl-2_2-6-remo-bicyclo [3.1.0] hexane_4,6_dicarboxylic acid difluorene Ester (0.5 g, 1.9 mmol, Preparation 4). Purification by PC_TLC (ethyl acetate / hexane) gave 0.94 g (79%) of the title compound. [Cheng = + 4 (c = 1.〇〇, CH3OH). lE NMR (300 MHz, CDC13) δ 1.44 (9H, s), 1.56 (9H, s), 2.44 (1H, t, J—4.0 Hz), 2.88 (1H, d, J = 14.7 Hz), 2.98 (1H , dd, J = 4.4 7.3 Hz), 3.04 (2H, d, J = 7.3 Hz), 3.38 (1H, dd, J = 4.0, 7.3 Hz) 3.77 (3H, s), 3.83 (3H, s) , 4.11 (1H, d, J = 14.3 Hz), 4.22-4.29 (1H, app q, J = 7.3 Hz), 4.92 (1H, bd, J = 7.7 Hz), 7.07 (1H, bs), 7.1-7.26 (4H, m). HRMS estimates C28H38N20i2SNa, 649.2043. Found: 649 2000. Preparation of 15 (lR, 4S, 5S, 6S) -4-,,,,, (3 '-_ dioxanyl ,, yl-2'S-,,,,,, dioxanylcarbonyl Group) -2,2-diketo-2λ6-fluorenyl-bicyclo "3.1 .〇1 hexane-4,6-dimiao_dimethyl ester 85435 -46- 200400815 h3co:

Prepared according to the general procedure A, using 3_acetoxy_23_ (third butoxycarbonylamino) propionic acid (0.25 g 'mmol of mol, preparation 44) and (1R, 4S, 5S, 6S) _44 group -2,2-diketo-2λ6- 喽 -bicyclo [3. 丨 · 〇] hexane_4,6_dicarboxylic acid dimethyl ester (0.2 g '0.8 dam' prepared 4) except dmaP and Unused. Purification by PC-TLC (ethyl acetate / hexane) yielded 019 g (48 2%) of the title compound. [<= 24 (c = 1.0, CH3OH). NMR (300 MHz, CDC13) δ 1.47 (9H, s), 2.10 (3H, s), 2.51 (1H, t, J = 4.4 Hz), 2.99-3.07 (2H, m), 3.43 (1H, dd, J = 4.0, 7.3 Hz), 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). HRMS estimate CBHwNWnSNa, 515.1312. Found 515.1305. Preparation 16 (1 foot, 25, 4! 1, 5 foot. 6 phan-2-amino-4-fluorobicyclo "3.1.01 hexane-2.6-diphosphonic acid diethyl ester

9 85435 -47- 200400815 p- (111,28,411,511,611) -2-amino-4-fluorobicyclo [3.1.0] hexane-2,6-dimetanoic acid (14.45 g, 71.12 millimolar, US Patent No. 5,958,960) was slurried in 202 ml of absolute ethanol at room temperature, and sulfite St dichloride (26 ml '356 mmol) was added dropwise over 20 minutes. The slurry is heated to reflux and allowed to stir for another 3 hours. Let it cool at room temperature and stir overnight. The resulting solution was concentrated to a residue under vacuum, then diluted with 136 ml of ethyl acetate, and treated with 306 ml of a 10% aqueous solution of sodium intact for 15 minutes, and vortexed by hand so that the final pH was '10. The layers were separated and the aqueous layer was washed with ethyl acetate (1x136 mL). The combined organic extracts were washed with brine (1 X 136 liters), dried (MgS04), filtered, and concentrated under vacuum to yield 17.07 g (93%) of the title compound as a white solid. [T 3 = + 20.37o (c = 1-l, MeOH). m.p. = 64-66 ° C. lU NMR (400 MHz, CDC13) δ 1.28 (3H, t, J = 7.3 Hz), 1.31 (3H, t, J = 6.8 Hz), 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, 14.1 Hz), 4.24 (2H, φ dq, J = 1.5, 7.3 Hz), 5.33- 5.52 (1H, m). Analysis estimates c12H18FN04: C, 55.59; H, 7.00; N, 5.40. Found: C, 55.29; H, 6.75; N, 5.45. &gt; MS (ES) m / z found 260.3 [M + H] +. Preparation 17 (1 £, 25,4 feet, 51_611) -2- 『2'8- (Third butane carbonyl amine its 鈥 醯 醯 醯 1_cryl-4 -fluorobicyclo『 3.1.01hexane- 2,6-di # acid diethyl vinegar 85435 -48-

200400815 To a solution of N-Boc-L-alanine (38.62 g, 204 mmol) in 396 ml of dichloromethane at -22 ° C and nitrogen was added N-methylmorpholine (22.44 ml, 204 Mmol), followed by isobutyl chloroacetate (26.48 ml, 204 mmol) added dropwise over 15 minutes, so that the reaction temperature does not exceed -18 ° C. Make the raw-into a thin slurry at -20. (: Stirring for 30 minutes, at this time (1R, 2S, 4R, 5R, 6R) -2-amino-4-fluorobicyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester ( 49.46 grams, 191 millimoles, Preparation 16) A solution of 247 ml of dichloromethane was added over 40 minutes so that the reaction temperature did not exceed -16 ° C. Once it was completely added, the reaction was removed from the cooling bath and left in the environment Stir at temperature for 70 minutes. At this time, the reaction temperature reaches 15t, and the color becomes light orange. The reaction is treated with 408 ml of 1 N HC1, stirred for 5 minutes, and then the layers are separated. The organic layer is washed with a saturated aqueous sodium hydrogen acetate solution ( 1x408 ml), dried (NasSCU), filtered, and concentrated under vacuum to produce a white foam (88.16 g). Analytical estimates of C20H31FN207 * 0.1 CH2C12: C, 55 · 00, H, 7.16; N, 6.38. Found: C, 55.18; H, 7.18; N, 6.49. MS (ES) m / z 431.3 [M + H] +, 331.2 [M + H-Boc] +. Preparation 18 〇_Ε, 28,4K · ， 5Κ · ， 6Κ) -2- | ~ 2υ ^ Triglyceride_ ^ Amino group amine 1 amine 4-fluorobicyclo 3 · 1 hexane-2.6-di-tae &lt; 85435 -49- 200400815

F

(111,28,411,511,61〇-2- [2 | 3- (Third-butoxycarbonylamino) propylfluorenyl] amino-4-fluorobicyclo [3 · ι · 〇] hexane_2, A solution of diethyl 6-dicarboxylate (8816 g, 191 mmol) was prepared in 238 ml of THF, and 238 ml (477 mmol) of 2 N sodium hydroxide was added at room temperature. The mixture was stirred vigorously at room temperature for 2.5 hours. At this point the reaction was homogeneous. The mixture was diluted with 238 ml of third butyl methyl chain, followed by mixing and layering. The aqueous layer was further diluted with 238 ml of water, and the particulate matter was removed by filtration. The solution was treated with concentrated HC1 (42.9 ml '515 mmol) for 30 minutes, and the title compound was seeded as needed, and stirred for 1 hour. The resulting slurry was filtered, washed with water (2 x 100 ml), and 45 C It was dried under vacuum for 40 hours to produce 72.2 g of the title compound as a white solid. A portion of the solid was stirred with 490 ml of acetone (69 · 5 g over a period of 1 hour to produce a dilute solution; filtered and washed with acetone (2χ 100 ml). The filtrate was concentrated under vacuum to a white foam, which was dried under vacuum at 45 ° C for 16 hours to yield 61.8 g (86% Corrected with 12% wt / wt acetone) The title compound. This material can be used in Examples 14-1 8 without clocking. Preparation 19,6. ) A bicyclic amine [3 〇1 hexane-2,6-dimiao acid diethyl ester 85435 -50- 200400815

pC02CH3CH3 NHBoc p- (18,28,48,511,611) -2- (third butoxycarbonyl) amino-4_hydroxybicyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester (1.50 G, 4.20 mmol, us 5,958,960), pyridine (0.365 ml, 4.62 mmol) and DMAP (0.513 g, 4.20 mmol) in dichloromethane (40 ml) under nitrogen Add acetic anhydride (0.5 to 14 ml, 5.0 mmol). It was stirred at room temperature for 16 hours, diluted with dichloromethane, and poured into a 10% aqueous citric acid solution (50 ml). The organic layer was washed with water (50 ml) and brine (50 ml). Drying over MgS04, filtering and concentrating in vacuo gave the title compound as a white solid (1.295 g, 75%). LCMS: m / z 400 [M + H] + and m / z 300 [M + H-CO / Bur @RT 1.39 minutes. Preparation of 20_ (15,28,48,511,61〇-4-acetamidine 1 and its 2-aminobicyclic "3.1.01, school-7 6-di # base diethyl acetate_

(18,28,43,511,6 feet) -4-ethoxy-2- (third-butoxycarbonyl) aminobicyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester ( 1.25 grams of '3.13 millimoles, Preparation 19) A solution of 95% TFA in dichloromethane (60 mL) was stirred under nitrogen at room temperature for 5 minutes. After this time the TFA / dichloromethane was removed under vacuum. The crude product 85435-51-200400815 was dissolved in a suspension of NaHC03 (1.00 g) in dichloromethane (50 ml) and stirred for 30 minutes. The filtered suspension 'was washed with dichloromethane (3 x 25 ml) and concentrated under vacuum to give 916 mg (98%) of the product as a yellow oil. LCMS: m / z 300 [M + H] + @Rt 0.92 minutes. Preparation of 21 (1S, 2S, 4S, 5R, 6R) -4-ethoxy-2- "2, SJ", its trimethyl group 1-amine, a bicyclic f 3.1 Wyatt _ 2.6-Diboronic Acid Diethyl Brew

Η ΝΗ 0

P- (lS, 2S, 4S, 5R, 6R) -4-ethoxy-2-aminobicyclo [3 · 1 · 〇] hexane-2,6-dicarboxylic acid diethyl ester (0.8 0g, 1.94 millimoles, Preparation 20) and a suspension of BOCL-Ala (0.477 g, 2.52 millimoles) in anhydrous dichloromethane (50 ml), and EDC (0,519 g , 2.72 mmoles), HOBt (0.3 14 g, 2.32 mmoles), catalytic DMAP (0.024 g, 0.19 mmoles) and triethylamine (1.08 ml, 7.76 mmoles). After stirring at room temperature for about 15 minutes, the original white suspension was completely dissolved. The reaction was stirred for 16 hours, diluted with dichloromethane, and washed with saturated NaHC03 aqueous solution (50 ml), i.on HC1 aqueous solution (3 x 20 ml), and saturated brine (40 ml). The organic layer was dried over MgS04, filtered, and concentrated under vacuum. The resulting crude amidine was purified by column chromatography using (4: 1) a mixture of ethyl acetate and hexane as the eluent 'to produce 638 mg (75%) of the product as a white solid. 85435 • 52- 200400815 ιΈί NMR (CD3〇D) *: δ 5.05 (1Η, d, 5.7 Hz), 4.09 (2H, q, 7.3 Hz), 4.02 (2H, q, 7.3 Hz), 4.01-3.94 (1H , m), 2.65-2.59 (2H, m), 2.38 (1H, d, 14 Hz), 2.10-2.03 (3H, m), 1.91 (3H, s), 1-78 (1H, dd, 5.9 Hz, 16 Hz), 1.59 (1H, brs), 1.34 (9H, s), 1.16 (6H, 2xt, 7.3 Hz); * NB exchangeable protons are not observed for NMR = 2. LCMS: m / z 471 [M + H] + and m / z 371 [M + H-CO / Bur @ RT = L30 minutes.

Rf 0.5 0 (80% ethyl acetate ·· heptane). Preparation of 22,8,28,48,5 feet, 61〇-2-amino-4-hydroxy-1,2-difluorene "3.1.01 忮 -2 &gt; 6- ~ ^^ diethyl acid

H3CH2C〇2C

(13,23,43,511,611) -2-Third-butoxycarbonylamino-4_hydroxy_bicyclo [3 · 1_0] Hex 'pit-2,6-dicarboxylic acid diethyl ester (50 g 14 mM, US ^ cooled from a solution called acetic acid in ethyl acetate to aquatic muscle ⑷ ⑷ steamed into the solution until saturation 'and stirred for 0.5 hours. Allow the reaction to warm to room temperature, Stir for another 1 hour. Concentrate the reaction mixture and pour between ethyl acetate and Η. Treat the aqueous layer with NaHC03 (aq) and extract with ethyl acetate. Dry the organic layer with k: 2C03 and concentrate to 22 g (59 7%) of white solid particles were generated. 85435 • 53 · 200400815 [α ^ =-30 · 8 (c = 0.52, CH3OH). 'H NMR (300 MHz, CDCl3) δ 126 (3H, t , Wind 33 Hz), i% (3H, t, 7.33 Hz), 1.60 (1H, dd, J = 5.87, 15.40 Hz), 1.64 (1H, t, J = 2.93 Hz), 2.27 (2H, bs), 2.15-2.29 (3H, m), 3.93 (1H, bs), 4.13 (2H, q, J = 6.97 Hz), 4.27-4.36 (3H, m). Analysis and estimation Cl2H19N05: C, 56.02; H, 7 · 44; N, 5.44. Measured values: c, 55.75; H, 7.36; N, 5.40. MS (ES) m / z 258.1 [M + H] +. ^ 23 Bile private call 511, 61 ^ #-third Jiamou neck-B-B please, -4-Hai _- bicyclo [3.1.01 has: two Miao diethyl

Prepared according to general procedure A, using Boc-glycine (458 mg, 2.62 mmol, Aldrich) and (13,23,48,511,611) -2-amino-4-hydroxy-bicyclo [3.1. 0] Hexane-2,6-dicarboxylic acid diethyl ester (450 mg, 1.75 mmol, Preparation 22). Purified on 35 g of silica, eluting with a gradient of 50/50 to 20/80 hexane / ethyl acetate. Yield: 560 mg (77%). lH NMR (400 MHz, CDC13) δ 1.28 (3Η, t, J = 7.3 Hz), 1.29 (3H, t, J = 7.3 Hz), 1.48 (9H, s), 1.57 (2H, m), 2.21 (1H , m), 2.41 (1H, dd, J = 2.9, 5.8 Hz), 2.81 (1H, d, J = 15.6 Hz), 3.76 (1H, d, 85435 -54- 200400815 J = 5.8 Hz), 4.10-4.18 (3H, m), 4.26 (2H, q, J = 7.3 Hz), 4.33 (1H, m), 5.10 (1H, bs), 6.86 (1H, bs). Analytical estimates of C19H300N2 08: C, 55.06; H, 7.30; N, 6.76. Found: c, 55.24; H, 7.50; N, 6.76. MS (ES) m / z 415.2 [M + H] +, 437.2 [M + Na] +. Preparation 24 Third butyl_oxoamine amine 3, _methyl butyl _ cyano) -4- and a -bicyclo [3.1.01hexan-2.6-Ⅱ # 醢 二 r, 艏

Prepared according to the general step A, using Boc-L-valine acid (569 mg, 2.62 millimoles' Sigma) and (^^ Lu / Lu Yichi / Phenylamido-cardiacyl-bicyclo ^ 丄 ... Diethyl-2,6-dicarboxylic acid (450 mg, 1.75 mmol, Preparation 22). The crude material was purified on 35 g of silica with a gradient of 70/30 to 20/80 hexane / ethyl acetate. Dissolution. Yield: 694 mg (87%) of a white foam. * H NMR (400 MHz, CDC13) δ 0.95 (3Η, d, J = 6.8 Hz), 1.0 (3H, d, J = 6.8 Hz) , 1.29 (6H, t, J = 7.3 Hz), 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-15.6 Hz), 3.86 (1H, dd, J = 6.3, 8.8 Hz), 4.14 (2H, q, J = 7.3

Hz), 4.21-4.32 (4H, m), 5.00 (1H, d, J = 8.3 Hz), 6.63 (1H, s). 85435 -55- 200400815 Analyzed and estimated C22H36N208: c, 57.88; 95, 7.95; N, 6.14. Found: C, 57.87; 87, 8.03; N, 6.12. MS (ES) m / z 457.2 [M + H] +, 479.2 [M + Na] +. Carbamide, amine, _4 dagger, _, _, _ _ _ _). -4- benzyl-bicycloalkane 2 6_ dimiao acid diethyl

Prepared according to general procedure A using bocL-leucine (606 mg, 2.62 mmol, Chemlog) and (13,28,48,511,611) -2-amino-4-hydroxy-bicyclo [3.1.0] Diethyl hexane-2,6-dicarboxylic acid (450 mg, 1.75 mmol, Preparation 22). The crude product was purified on 35 g of silica and eluted in a gradient of 70/30 to 2O / 80 hexane / ethyl acetate. Yield: 689 mg (84%) of a white foam. ! H NMR (400 MHz, CDC13) δ 0.96 (6Η, m), 1.28 (3Η, t, J = 7.3 Hz), 1.29 (3H, t, J = 7.3 Hz), 1.46 (9H, s), 1.45- 1.73 (5H, m), 2.20 (1H, m) 5 2.42 (1H, m), 2.80 (1H, d, J = 16.1 Hz), 4.07-4.35 (7H, m), 4.81 (1H, bd), 6.86 (1H, bs). Analysis estimates C23H38N2O8 * 0.1 H20: C, 58.48; H, 8.15; N, 5.93. Found: C, 58.22; H, 7.94; N, 5.92. MS (ES) m / z 471.2 [M + H] +, 493.2 [M + Na] +. 85435 -56- 200400815 Preparation 26 (1_8,28,48,511, PAN | 1) :: 2- (2,8-Third oxocarbonyl certain amine-3,8-methyl certain-zhong | xi &amp; ) -4-hydroxy-bicyclo "3.1.01 hexane-2.6-di # acid dibenzoate

Prepared according to General Procedure A, using Boc-L-isoleucine (606 g, 2.62 mmoles' Aldrich) and (18,28,48,511,61〇-2-amino-4-hydroxy_bicyclo [ 3.1.0] Dihu-2,6-dicarboxylic acid diethyl ester (450 mg, 1,75 mmol, Preparation 22). The crude product was purified on 35 g of silica '70 to 30 to 20/80 Rhenium / ethyl acetate gradient dissociation. Yield: 731 mg (89%). LB NMR (400 MHz, CDC13) δ 0.92 (3Η, t, J = 7.3 Hz), 0.97 (3H, d, J = 6.8 Hz) , 1.17 (1H, m), 1.29 (6H, t, J = 6.8 Hz), 1.46 (9H, s), 1.53 (1H, dd, J = 5.8, 15.6 Hz), 1.58 (1H, t5 J = 2.4 Hz ), 1.74 (1H, m), 1.83 (1H, m), 2.20 (1H, m), 2.43 (1H, m), 2.80 (1H, d5 J = 15.6 Hz), 3.88 (1H, dd5 J = 7.3, 8.8 Hz), 4.12-4.32 (6H, m), 4.98 (1H, d, J = 7.3 Hz), 6.60 (1H, s). Analysis and estimation C23H38N2O8 * 0.2 H20: C, 58.25; H, 8.16; N, 5.91 Measured values: C, 58.17; H, 8.11; N, 5.91. MS (ES) m / z 471.2 [M + H] +, 493.2 [M + Na] +. Preparation 27 £ 1 ^ 128748,511,611) -2- 『2 |-(2-Third-butoxycarbonyl group_ethamidamine） · 85435 o7- 200400815 —Yanqi-_ji ~ 111: .Jiji-difluorene『 3.1.〇1 Alkoxy -2,6 -: ^ [^ hydroxyalkyl acid

Prepared according to general procedure A, using Boc-Gly-Gly (474 mg, 2.04 mmol) and (18,28,48,511,6 feet) -2-amino-4-rokonyl-bicyclo [3.1.〇 ] Hexane-diethyl 2,6-dicarboxylate hydrochloride (400 mg, 1.36 mmol, Preparation 22, without basic treatment) with the following exceptions. DMAP is not used. Add one equivalent of triethylamine. The crude material was purified on 35 g of silica and dissolved in ethyl acetate. Yield: 517 mg (81%). [a] g = -18.18 (c = 0.55, MeOH). NMR (300 MHz, CDC13) δ 1.26 (3H, t, J = 6.8 Hz), 1.27 (1H, t, J = 7.3 Hz), 1.28 (3H, t, J = 6.8 Hz), 1.46 (9H, s), 1.59 (1H, t, J = 3.4 Hz), 1.62 (1H, dd, J = 6.3, 16.1 Hz), 2.21 (1H, dd, J = 2.9, 5.8 Hz), 2.46 (1H, dd, J = 2.4, 5.8 Hz), 2.73 (1H, d, J = 15.6 Hz), 3.78-3.91 (3H, m), 4.01-4.15 (3H, m), 4.24 (2H, q5 J = 6.8 Hz) 4.32 (1H, d, J = 5.8 Hz), 5.28 (1H, b), 6.87 (1H, bt, J = 4.9 Hz), 7.39 (1H bs). HRMS estimates C21H34N309, 472.2295. Found: 472.2303. HPLC: 16.755 minutes. Column: Symmetry C18, 3.5 microns, 4.6X150 mm, λ = 230 nM. Flow rate: 1 ml / min. Gradient: i0% _50% ACN / water contains 0.1% TFA over 25 minutes. -58- 33G 85435 200400815 Preparation 28 iia., 2S, 4S.5R.6RV2- "2,-(2S-Third-butoxycarbonyl-propionate, etc.) Π.1.01 Hexane-2,6-diacid acid

Prepared according to the general procedure A, using Boc-Ala-Gly (502 mg, 2.04 mmol) and (1S, 2S, 4S, 5R, 6R) -2-amino-4-quinyl-bicyclo [3.1.0 ] Hexane-2,6--diacetic acid diethyl ester hydrochloride (400 mg, 1,36 mmol, preparation 22 without basic treatment) 'with the following exceptions. Do not use DMAP. Add one equivalent of triethylamine. The crude material was purified on 35 g of silica and isolated with ethyl acetate. Yield: 500 mg (76%). [&lt; =-31.37 (c = 0.55, MeOH). 4 NMR (300 MHz, CDC13) δ 1.27 (3H, t, J = 6.8 Hz), 1.28 (3H, t, J = 6.8 Hz), 1.42 (3H, d, J = 6.8 Hz), 1.45 (9H, s ), 1.58 (1H, t, J 3.4 Hz), 1.64 (1H, dds J = 5.8, 15.6 Hz), 2.23 (1H, dd, J = 3.4, 5-8 Hz), 2.50 (1H, dd, J = 2.9, 6.3 Hz), 2.69 (1H, d, 1 = 15.6 Hz), ABq (2H, Va = 3 87, Vb = 3 98, Jab = 17i HZ, Jd = 63

Hz), 3.92 (1H, m), 4.07-4.16 (3H, m), 4.24 (2H, q, J = 7.3 Hz), 4-32 (1H, b), 5.08 (1H, b), 6.84 (1H , bt, J = 4.88 Hz), 7.14 (1H, bs) 0 S5435 -59- 200400815 HRMS Estimate C22H36N309, 486.2452. Found: 486.2451. Preparation of 29 nS, 2S, 4 ,,,,, 3,511,6Da) -2- (2 | -Third-Butanidine, a certain amine, -3, -Mo 1-min_risk_yl) -4-¾ -Second Ring "3.1.01 Hexagonal Burning"

C〇2CH2CH. ΗN was prepared according to the general procedure A, using bocL-phenylalanine (772 mg, 2.91 Kelvin) and (1S, 2S, 4S, 5R, 6R) -2-amino-4- -Bicyclo [3.1 0] Hexyl-2,6-hexanyldiethylstil hydrochloride (480 mg, 1.94 mmol, Preparation 22). The crude reaction mixture was concentrated, dissolved in dichloromethane and quickly filtered through a short crushed rubber plug to ethyl acetate / DCM (1: 1) and concentrated under vacuum to produce a yellow oil. The material was further purified by silica gel chromatography using 30% ethyl acetate / hexane to 80% ethyl acetate / hexane to yield 852 mg (87%) of the title compound. [4 = -23.66 (c = 0_93, MeOH) 〇 (H NMR (400 MHz, CD3OD) δ 1.22-1.30 (6H, m), 135 (9H, s), 1.59 (1H, t, J-2.9 Hz) , 1.70 (1H, dd, J = 5.5, 15,4 Hz), 2.06 (1H, m), 2.40 (1H, d, J = 15.4 Hz), 2.63 (1H, dd, J = 2.9, 5.9 Hz), 2.76

(1H, dd, J-9.2, 13.9 Hz), 4.08-4.33 (6H, m), 7.20-7.29 (5H m). MS found: 505.0 [M + Η] +. 85435 -60- 200400815 HRMS estimates C26H36N208, 505.2550. Found: 5205 2559. Preparation of 3〇_ tertiary butylamine (trimethylolamine-aminopyridine)-butylaminoamine 1-4-yl-bicycloπ 1 · 〇1 hexadecyl-2,6-diacoic acid two

Prepared according to general procedure A, using bocL-glutamine (trityl) _〇Η (1.40 g, 2.86 mmol) and (18,28,48,5 feet, 611) -2-amine Di-4-hydroxy-bicyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester (490 mg, 1.90 millimoles 'Preparation 22)', with the exception of not using DMAP. The concentrated crude reaction mixture 'was dissolved in dichloromethane, quickly filtered through ethyl acetate via a short plug of silica' and concentrated under true pressure to give a yellow oil. Further purification by gel chromatography was performed by dissociation with 30% ethyl acetate / hexane to 100% ethyl acetate / hexane to give 1.3 g (94%) of the title compound. [〇 ^ 3 = -21.28 (c = 0.94, MeOH). NMR (400 MHz, CD3OD) 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) = 85435 -61-200400815 MS found: 728.2 [M + H] +. HRMS estimates sf C41H49N3O9, 728.3547. Found: 728 3533. Preparation 31 (1S) 2S, 4S, 5R, Li Jin)-2 ~ (2'S, 6'-l-the third butyl-oxycarbonyl neck-a ^ 1! 1 ^^ _- Second Ring "3.1 .01 Hexane-2.6- Dimiaoxi 6-Guang

Prepared according to general procedure A, using Boc-L-lysine (Boc) -OH (9.10 mg, 2.63 mmol) and (18,23,43,5 feet, 611) -2-amino-4-hydroxyl -Bicyclo [3.1_0] hexane-2,6-dicarboxylic acid diethyl ester (450 mg, 1.75 mmol, Preparation 22), except that DMAP was not added to the reaction mixture. The crude reaction mixture was concentrated, dissolved in dioxane, quickly filtered through a short silica gel plug, ethyl acetate, and concentrated under vacuum to produce a yellow oil. Further purification of the substance 'using silica gel chromatography and dissociation with ethyl acetate / hexane (?: 1) yielded 800 ¾ (78%) of the title compound. [&lt; =-30.19 (c = 0.53, MeOH). NMR (400 MHz, CD3OD) δ 1.21-1.24 (6H, m), 1.26-1.77 (27H, m), 2.07 (1H, m), 2.42 (1H, d, J = 15.8 Hz), 2.59 (1H, dd , J = 2.6, 5.5 Hz), 3.03 (1H, t, J = 6.2 Hz), 3.99 (1H, m), 4.07-4.28 (5H, m). MS found: 586.1 [M + H] +. -62- 34 85435 200400815 HRMS estimate C28H47N301 (), 586.3340. Found: 586.3348. Preparation of 32 QS II! ^ 4m6R) -2- 〖2, · The third butoxyfluorenylamine _3, _ (tribenzyl _neck-methoxamine 1-4-hydroxy-double-coated "V 1 〇1 -2,6-diethyl diacetate

Ph Ph

Prepared according to the general procedure A using Boc-L-asparagine (trityl) -OH (1.35 g '2.84 mmol) and (18,28,48,5,6 feet) -2- Amino-4-hydroxy-bicyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester (476 mg, 1.85 mmol, Preparation 22), except that DMAP is not added to the reaction mixture . The crude reaction mixture was concentrated 'dissolved in dichloromethane' and quickly filtered through a short plug of silica gel using ethyl acetate and concentrated under vacuum to yield a yellow oil. It was further purified by silica gel chromatography using 20% ethyl acetate / hexane to 50% ethyl acetate / hexane to dissociate &apos; to yield 1.07 g (81%) of the title compound. [T 3 = -25_45 (c = 0.55, MeOH). ! H NMR (400 MHz, CD3OD) δ 1.19-1.26 (6H, m), 1.45 (9H, s), 1.61-1.69 (2H, m), 2.05 (1H, m), 2.44 (1H, d, J- 15.4 Hz), 2.51-2.72 (3H, m) 5 4.02-4.21 (4H, m), 4.25 (1H, d, J = 5.5 Hz), 4.36 (1H, dd, J = 4.8, 8.8 Hz), 7.18- 7.30 (15H, m). -63- 200400815 MS found 714.1 [M + H] +. HRMS estimates are 〇4〇47Ν307, 714.3391. Measured value: 714.3380 0 Preparation 33 Tertiary butyl 1-carbonyl amine -341- Tertiary butyl cyano-3'-yl) -Propanamido) -4 · 衮 -Di hen Diethyl-2,6-dicarboxylic acid

Prepared according to general procedure A, using BOC-L_tryptophan (Boc) _OH (118 g, 2.9 丨 mmol) and (18,28,43,511,611) _2_amino_4_hydroxy -Bicyclo [31〇] hexane-2,6-dicarboxylic acid diethyl alcohol (520 mg, 190 mmol, Preparation 22). The crude reaction mixture was concentrated, dissolved in dichloromethane, quickly filtered through a silica plug, using ethyl acetate, and concentrated under vacuum to give a yellow oil. Further purification of the shellfish by silica gel chromatography with ethyl acetate / DCM (1: 1) gave 12 g (92%) of the title compound. [α] 1) 3 :: =-12 · 5 (c = 0.96, MeOH). 4 nMR (400 MHz, CD3OD) s (18_127 (6h, called, i 34 (9H, s), 1.62 (1H, t, J = 2.9 Hz), 1.67 (9H, s) 5 1.74 (1H , dd, J = 5.9, 15.2 Hz), 2.07 (1H, m), 2.46 (1H, d,] = 15 · 7 Hz), 2 ㈣ (1H, d, J = 2 9, 5-9 Hz), 2.89 (1H, dd, J = 9.3, 15.0 Hz), 3.16 (1H, dd, 1 = 4.9, -64-33k 85435 200400815 15.2 Hz), 4.04-4.12 (2H, m), 4.18-4.24 (2H, m ), 4.27 (1H, d, J = 5.4 Hz), 4.43 (1H, dd, J = 5 „4, 9.3 Hz), 7.26 (2H, m), 7.52 (1H, s), 7.64 (1H, d, J = 7.3 Hz), 8 · 10 (1H, d, J = 8.3 Hz). MS found: 644.8 [M + ΗΓ, 666.8 [M + Na] +. HRMS estimated CwHmNsO !, 666.3002. Found: 666.2988. Preparation of 34 (im4S, 5R, 6R) _2- (r-th-tributyrin. 藕 miao · pubu, pyridine-2'S-carbonyl-m 某 v4-stupid— 棊 bicyclic "3 · 1 · 0 丄 丄Alkane_2.6-dicarboxylic acid diethyl ester

Prepared according to general procedure A using Boc_ £ _proline and (1S, 2S, 4S, 5R, 6R) -2-amino-4-¾yl-bicyclo [3.10] hexane_2,6_ Diethyl dicarboxylate (Preparation 22). The crude material was purified on 110 grams of silica in an ISCO system and was isolated in a gradient of 80% ethyl acetate / hexane to 100% ethyl acetate to yield 699 mg (84%) of the title compound. [α: β3 = -52 · 53 (c = 0.99, MeOH). ] H NMR (400 MHz, CD3OD) δ 1.26 (6H, m), 1.44 (9H, s), 1.62 (1H, m), 1.73 (1H, dd, J = 5.4, 15.3 Hz), 1.84 (1H, m ), 1.97 (1H, m), 2.07 (4H, m), 2.41 (1H, d, J = 15.3 Hz), 2.50 (1H, m), 2.59 (1H, m), 4.12 (2H, m), 4.20 (1H, q, J = 7.4 Hz), 4.27 (1H, d, J = 5.4 Hz). 85854 -65- 200400815 MS found. · 454.9 [M + H] +, 476.8 [M + Na] +. HRMS estimate C22H34N208A, 477.2213. Found: 477.2210. Preparation 35 〇_8,28,45,5 Good 61〇-2- | ~ 2 | 8-Third butyl succinate amine ^^ _ 4_ rod: butyl-orange green oxygen, base-benzyl)- Propylamino 1-4-benzyl-bicyclo 『3.1.01 hexadecor; -2,6-di # acid

The title compound was prepared according to the general procedure A, using 2S-third butoxycarbonylamino-3- (4-third butoxycarbonyloxy) -phenyl) -propionic acid and (18,28,43,511,611)- 2-Amino-4-mercapto-bicyclo [3.1.0] hexane-2,6-diacetic acid diethyl ester (Preparation 22). The crude material was purified on 110 g of silica in an ISCO system and was isolated in a gradient of 60% ethyl acetate / hexane to 90% ethyl acetate / hexane to yield 1.09 g (91%) of the title compound. Ton = -12 (c = 1.0, MeOH). NMR (400 MHz, CD3〇D) δ 1.25 (6H, m), 1.37 (9H, s), 1.52 (9H, s) 9 1.68 (1H, dd, J = 5.4, 15.3 Hz), 2.06 (1H, m ), 2.38 (1H, d &gt; J = 15.3 Hz), 2.64 (1H, m), 2.79 (1H, dd, J = 8.4, 13.9 Hz), 85435 -66- 200400815 3.05 (1H, dd, J = 5.4, 13.9 Hz), 4.20 (6H, m), 7.04 (2H, d, J = 8.4 Hz), 7.26 (2H, d, J = 8.4 Hz). MS found: 621.8 [M + H] +, 643.8 [M + Na] +. HRMS estimate C3iH44N2O &quot; Na, 643.2843. Found: 643.2845. Preparation of 36 nS, 2S.4S.5R, 6R) -2- (2, S-Third-butane hydrocarbamate_4 dagger a certain sulfur a certain _ butamidinyl group) -4- meridian-second soil 3.1.01 hexane-2,6-di # acid diethyl vinegar

The title compound was prepared according to general procedure A, using (Boc-L-methionine) and

(18,23,43,5 feet, 610-2-amino-4-hydroxy-bicyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester (Preparation 22). The crude material was Purified on 110 g of silica and eluted on a ISC0 system using a gradient of 80% ethyl acetate / hexane to 100% ethyl acetate to produce 1.0 g (92%) of the title compound. Post = -30 (c = 1.0, MeOH). * H NMR (400 MHz, CD3OD) 6 1.26 (6H, m), 1.43 (9H, s), 1 34 (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 found: 488.8 [M + H] +, 510.8 [M + Na] +. HRMS estimated C22H36N208SNa, 511.2090. Found. 5i [2071. -67-

339 85435 200400815 Preparation 37 £ 1§., 28,48,5 feet, 61): 2- (2 | 8-Third 1 some carbonyl amine _ propyl 醯 neck very much 4_ ashamed; 餐:. Meal 1 ~ 3.1.〇1 Burning: 2,6-two miaoling two λ 艏

Prepared according to general step A, using commercially available N_Boc_ (L) _alanine and (lS, 2S, 4S, 5R, 6R) -2-amino-4- # yl-bicyclo [3.1.0] hexane Alkane-2,6_diacetic acid diethyl alcohol (Preparation 22). The reaction mixture was refluxed overnight and purified by preparative hplC on a 1 x 500 g SiO 2 column (10% ethyl acetate / hexane to 100% ethyl acetate). This produced 3.0 grams (90% &apos; 7.00 millimoles) of a white foam. [«^ =-32.31 ° (c = 0.37, CHC13). * Η NMR (300 MHz, CDC13) δ 1.28 (6H, t, J = 7.3 Hz), 1.34 (3H, d, J = 7.3 Hz), 1.46 (9H, s), 1.51-1.58 (2H, m), 1.64 (1H, s), 2.19 (1H, dd, J = 3.3, 6.2 Hz), 2.40 (1H, dd, J = 2.9, 6.2 Hz), 2.80 (1H, d, J = 15.8 Hz), 4.10-4.36 (6H, m), 4.92 (1H, bs), 6.94 (1H, bs). Analysis estimates C2QH32N2Os * 0.1 H20: C, 55 · 83; H, 7.54; N, 6.51. Found: C, 55.57; H, 7.64; N, 6.44. MS (ES) m / z 429.2 [M + H] +, 329.1 (M-Boc) +. Preparation 38 ^ 28,48,5 ^ 610-2- (23-Amine-Propanylamine) -4-Methyl-Bicyclo-68- 34 (5 85435 200400815 "3.1.01 Hexane-2 · 6-dicarboxylic acid ^ manganate

Add (1S, 2S, 4S, 5R, 6R) -2- (2'S-third butoxycarbonylamino-propanamido) -4-hydroxy-bicyclo [3.1.0] hexane-2,6- A solution of diethyl dicarboxylate (2.95 g, 6.88 mmol, Preparation 37) in ethyl acetate (30 ml) was charged with anhydrous HC1 gas at 0 ° C until the solution was saturated with HC1. The resulting reaction mixture was stirred at 0 ° C until the reaction was judged complete by TLC. Fill the reaction mixture with 30% for 30 minutes to remove excess HC1 gas. The resulting suspension was concentrated to dryness under vacuum to yield 2.47 g (98%, 6.78 mmol) of the desired amine hydrochloride. No further purification was required. [Ton 3 = -28.0. (C = 0.50, MeOH).

lH NMR (300 MHz, CDC1) δ 1.24 (3H, t, J = 7.0 Hz), i.2s (3H t 'J = 7.3 Hz), 1.50 (3H, d, J = 7.3 Hz), 1.61 (1H, t, J = 2 9 Hz) 1.76 (1H, dd, J = 5.9, 15.8 Hz), 2. · 10 (1H, dd, J = 3.3, 5.9 Hz) '2.43 (1H, d, J = 15.4 Hz) , 2.60 (1H, dd, J = 2.9, 6.2 Hz), 3 900Hz, q, J = 7.0, 13.9 Hz), 4.15 (2H, q, j = 7.3 Hz), 4.14-4.31 (3h m). , Do not analyze estimates C15H24N2O6 * HCN0.7 H20: C, 47.73; H, 7 05. N 7.42 ° Found: C, 47.96; H, 6.91; N, 7.04. , '85435 -69- 200400815 MS (ES) m / z 329.1 [μ + Η] + 〇 Preparation 39 Q third butyl amine aminopropyl amidinyl amine) _propyl amine amino 1-4 diyl -Bicyclic `` 3.1.01 alkane 26_26_ dimiao acid diethyl ester

Prepared according to general procedure A, using (18,28,43,511,611) -2- (2,3-amino-propanamido) -4-hydroxy-bicyclo [3.1.0] hexane-2,6 -Diethyl dicarboxylate hydrochloride (0.2 g '0.55 mmol, Preparation 38) and Boc- (L) -alanine (0.16 g, 0.82 mmol) with the exception of not using DMAP. Purification with PC-TLC (ethyl acetate / hexane) gave 0.13 g (47.3%) of the title compound. Κβ3 = -46 · 2 (c = 0.52, CHC13). lR NMR (300 MHz, CDC13) δ 1.27 (3H, t, J = 7.0 Hz), 1.28 (3H, t, J = 7.0 Hz), 1.36 (3H, d, J = 7.0 Hz), 1.37 (3H, d , J = 7.0 Hz), 1.44 (9H, s), 1.58-1.65 (2H, m), 2.19 (1H, dd, J = 3.0, 5.9 Hz), 2.46 (1H, dd, J = 2.6, 5.9 Hz) , 2.70 (1H, d, J = 15.4 Hz), 4.09-4.33 (7H, m), 4.48 (1H, app p, J = 7.0 Hz), 5.05 (1H, bd, J = 6.6 Hz), 6.79 (1H , bd, J = 7.7 Hz), 7.26 (1H, s). HRMS estimates C23H38N309, 500.2608. Found: 500.2598. Preparation 40 85435 -70- 200400815 (18,28.45.5 anion, 610-2- 『2,8- (2-third butane #a certain amino-acetamidoamine and octapropane a certain amino 1-hydroxy Sodium-bicyclic Π 丄 〇1 hexane-2,6-dicarboxylic acid diethyl ester

Prepared according to general procedure A, using Boc-glycine (0.29 g, 1.6 mmol) and (18,28,48,511,611) -2- (2 | 8-amino-propanamido) -4- Hydroxy-bicyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester hydrochloride (0.4 g, 1.1 mmol, Preparation 38) The exception is the absence of DMAP. Purification by PC-TLC (ethyl acetate / hexane) yielded 0.14 g (26.2%) of the target compound. [Cheng = -14 (c = 1.00, CDC13) 〇'H NMR (300 MHz, CDC13) δ 1.27 (3H, t, J-7.3 Hz), 1.29 (3H, t, J = 7.3 Hz), 1.38 (3H , d, J = 7.0 Hz), 1.45 (9H, s), 1.57-1.65 (2H, m), 2.19 (1H, dd, J = 3.3, 5.9 Hz), 2.44 (1H, dd, J = 2.9 , 5.9 Hz), 2.74 (1H, d, J = 15.8 Hz), 3.70-3.86 (2H, m), 4.08-4.34 (6H, m), 4.56 (1H, app p, J = 7.0 Hz), 5.31 ( 1H, bs), 6.88 (1H, bd, J = 7.0 Hz), 7.50 (1H, s). HRMS estimates C22H36N309, 486.2452. Found: 486, 2444. Preparation 41 (18,28,48,511,611) -2- "2 |-(28-Third-butanylphenoxy-4-methyl-pentamylamino) -propanylamino 1-4- Hydroxyl-bicyclo [3.1.01 hexane-2,6-di-chicoic acid diethyl ester-71-34; 85435 200400815

Prepared according to general step B, using commercially available N_Boc_ (L) _leucine monohydrate and (1S, 2S, 4S, 5R, 6R) -2- (2'S-amino-propanamido) _4 -Hydroxy-bicyclo [3.1 · 〇] hexane-2,6-dicarboxylic acid diethyl ester hydrochloride (0.2 g, 0.55 mmol). Purified with pc-TLC, 4 mm SiO 2 rotor (10% ethyl acetate / hexane to 1000 /. Ethyl acetate). Yield 0.54 g (62/100, 100 mmol) white foam. [〇 ^ 3 = -61.2o (c = 0.49, MeOH). 4 NMR (300 MHz, CDC13) δ 0.93 (3H, d, J = 6.2 Hz), 0.94 (3H, d, J = 6.2 Hz), 1.25 (3H, t, J = 7.3 Hz), 1.27 (3H, t , J = 7.3 Hz), 1-36 (3H, d, J = 7.0 Hz), 1.43 (9H, s), 1.53 (1H, d, J = 9.5 Hz), 1-55-1.69 (5H, m) , 2.18 (1H, dd, J = 2.9, 5.9 Hz), 2.44 (1H, dd, J = 2.9, 6.2 Hz), 2.70 (1H, d, J = 15.8 Hz), 4.06 (1H, bs), 4.13 ( 4H, q, j = 7.3 Hz), 4.18-4.28 (2H, m), 4.31 (1H, d, J = 5.9 Hz), 4.41-4,45 (1H, m), 4.85 (1H, bs), 6.57 (1H, d, J = 7.3 Hz), 6.97 (1H, bs) o Analyzed and estimated C26H43N3O9 * 0.1 H20: c, 57.46; H, 8.01; N, 7.73. Found: C, 57.18; Η, 8.00; N, 7.64. 3.44 85435 -72-200400815 HRMS estimate C26H43N309 [M + Na +] 564.2897. Found: 564.2922. Preparation 42 (1 S, 2 ____ § ___ a4S, 5R, 6E) -2-Ll ^ §I..H group) -Propanylamino group 1-4-Changou-Difluorene "3.1.01 2-2,6 -二 # Acid diethyl ester

0 CK is prepared according to general step B, using commercial n-BOC- (L) -valine acid and (13,23,48,511,611) -2- (2 | 8-amino-propanylamino) 4-Hydroxy-bicyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester hydrochloride (02 g, 055 mmol, Preparation 38). Purified with PC-TLC, 4 mm SiO 2 rotor, (10% ethyl acetate / hexane to 67 ° / ethyl acetate). Yield 0.36 g (43%, 0.68 mmol) white foam. [a] 2D3 = -65,5o (c = 0.58, MeOH). NMR (300 MHz, CDC13) δ 0.91 (3H, d, J = 7.0 Hz), 0.97 (3H, d, J = 6.6 Hz), 1.27 (3H, t, J = 7.3 Hz), 1.28 (3H, t, J = 7.0 Hz), 1.37 (3H, d, J = 17.3 Hz), 1.44 (9H, s), 1.54-1.61 (3H, m), 2.12-2.20 (2H, m), 2.42 (1H, dd, J = 2.6, 5.9 Hz), 2.70 (1H, d, J = 15.4 Hz), 3.90 (1H, t, J = 6.6 Hz), 4.13 (2H, q, J = 7.0 Hz), 4.20-4.29 (2H, m ), 4.31 (1H, d, J = 5.9 Hz), 4.40-4.48 (1H, m), 85435 _73 · 200400815 4.93 (1H, bs), 6.41 (1H, d, J = 7.3 Hz), 6.91 (1H, bs). Analysis estimates C25H41N3O9 * 0.2 CH2C12: C, 55.58; H, 7.66; N, 7.72. Found: C, 55.76; H, 7.27; N, 7.49. MS (ES) m / z 528.3 [M + H] +. Preparation 43 ^ 25,48,511,6? 0-2- (2 $ -Ethylamido-propylammonium amine) -4- hydrazine-bicyclic "3.1.01 hexane-2,6-dicarboxylic acid Diethyl ester

(1S, 2S, 4S, 5R, 6R) -2- (2, S-amino-propanamido) -4-hydroxy-bicyclo [3.1.0] hexane-2,6-dicarboxylic acid di A solution of ethyl acetate hydrochloride (0.30 g, 0.82 mmol, Preparation 38) in CH2C12 (30 ml), triethylamine (0.16 g, 1.6 mmol) was added at 0 ° C, and acetamidine was added. Chlorine (0.10 g, 1.18 mmol). The reaction mixture was allowed to stir and warm overnight. The reaction mixture (700 mL) was diluted with ethyl acetate, and washed sequentially with NaHS04 and brine. The organic layer was dried over magnesium sulfate, purified with a PC-TLC, 4 mm SiO 2 rotor (10% ethyl acetate / hexane to 100% ethyl acetate), yielding 0.24 g (79%, 0.65 mmol) ) White foam. [«^ = -48 ° (c = 0.5, MeOH). * H NMR (300 MHz, CDC13) δ 1.27 (3H, t, J = 7.3 Hz), 1.28 (3H, t, J = 7.3 Hz), I.37 (3H, d, J = 7.0 Hz), 1.55- 1.62 (2H, m), I. 99 85435 -74- 200400815 (3H, s), 2.19 (1H, dd, 1 = 3.3, 6.6 Hz), 2.40 (1H, dd, J = 2.6, 5.9 Hz), 2.77 d? j = i5 § Hz), 4.07 (2H, q, J = 7.0 Hz), 4.21- 4.31 (2H, m) 5 4.33 (1H, d, J = 6.2 Hz), 4.43-4.47 (1H, m) , 6.13 (1H, bs), 7.06 (iH, bs). Analysis estimates C17H26N207: C, 55.13; H, 7.08; N, 7.56. Found: C, 55.05; H, 7.12; N, 7.29 ° HRMS estimate C17H26N207 [M + Na +] 393.1638. Found: 393.1644. Preparation 44 Turtleoxy-2S- (third butane # yl) propionic acid

NHBoc N methylmorphine (28 g, 27.2 mmol) and di-tert-butyl dicarbonate (5.8 g, 25.2 mmol) were added to 0-acetamido_1 ^ serine (37 G, 252 pen moles) in a 1: 1 solution of diazane: water. The reaction was stirred for 24 hours and then partitioned between ethyl acetate and water. The aqueous phase was extracted with ethyl acetate and the organic phase was discarded. Adjust the pH to 0-1 with NaHSCU aqueous solution. The aqueous layer was extracted with ethyl acetate, dried over NazSCU, filtered, and concentrated under vacuum. Purification by flash chromatography yielded 2.6 g (41.7%). This material can be used in Preparation 15 without further identification. Preparation of 45 ϋR) -3- (brother f Ί1 ... Qisai some base group) -3- (third butoxy, garlic and even _ ring "3.1.01 hexane 85435 -75- 200400815 co2c (ch3) 3 NHBoc A solution of tertiary butyl methylmalonate (129 g, 0.75 mol) in THF (3 85 mmol) was added to LiH (14.9 g, 1.875 mol) in THF (900 ml) and Ν, Ν -A slurry of dimethyl propylene urea (DMPU, 155 g, 1.2 mol) over 30 minutes while maintaining the temperature at 0-5 ° C. The reaction mixture was heated to 65 ° C, and cis-1 was added. A solution of 4,4-dichloro-2-butene (95%, 100 g, 0.8 mol, 1.08 equivalent) in THF (100 ml) for 5.5 hours at a temperature maintained at 63-67 ° C. The reaction was stirred at 65 ° C for 4 hours. The reaction mixture was treated with water / MTBE to produce 1- (methoxycarbonyl) -1- (third butoxycarbonyl) cyclopentane-3.ene. 'H NMR (300 MHz , CDC13) δ 5.57 (s, 2H, CH = CH), 3.71 (s, 3H, CH3), 2.95 (s, 4H, 2CH2), 1.42 (s, 9H, C (CH3) 3). IR (thin film) 1734 (C = 0), 1646, 1268, 1149 cm-1. Cool the reaction solution containing 1- (methoxycarbonyl) -1- (third butoxycarbonyl) cyclopentane_3_ 晞 to 10-15 ° C, then increase to 1 N NaOH (1.3 liters, 1.3 moles) in a cooled solution (10-15 ° C) for 30 minutes. Stir the reaction solution at 25 ° C for 24 hours, and then follow up by GC analysis. When the reaction hydrolysis is complete, MTBE (645 ml) was added to the reaction mixture and stirred for 5 minutes. The layers were allowed to settle and separate. The organic layer was discarded. 1.5 M NaHSCU solution (1470 ml) was added to make the aqueous layer acidic (pH 2-3). MTBE (1 · 3 liters) and separated. The aqueous layer was extracted with MTBE (385 ml), and the combined organic layers were washed with 5% LiC1 solution. The organic layer was concentrated under vacuum and diluted with heptane (78 mm). Solution -76 -34S 85435 200400815 Concentrated to about 500 ml 'The resulting slurry is stirred at ambient temperature for 丨 hours. The filtered solid' is washed with heptane (250 ml) and then dried at 35. (: under suction and dried to produce 103.29 g (61%) Yield) of carboxylic acid) _ (3rd-butoxycarbonyl) cyclopent-3-ene as a white solid. Mp = 119 ° C. LH NMR (300 MHz, CDC13) δ 5.61 (s, 2Η, CH = CH) , 3.00 (s, 4H, 2CH2), 1.46 (s, 9H, C (CH3) 3). IR: 3800-3000 (br, COOH), 1741 (C02R), 1705 (C02H), 1283), 1149 cm-1. 1- (Acrylic acid) -1- (third butoxycarbonyl) cyclopentane-3-fluorene (50 g, 236 mmol) dissolved in 850 ml of 70:30 toluene: MTBE, 1 liter under nitrogen The flask was mechanically stirred. To the stirred reaction mixture was added thionyl chloride (33.6 g '283 millimoles, 1.2 equivalents), and the temperature was maintained at 23. (: Lower. The reaction solution was cooled to 0-5 ° C, and then triethylamine (32.2 g, 3 18 mmol, 1.35 equivalents) was added dropwise for 30 minutes. The reaction mixture was warmed to 23 ° C and stirred 1 hour. The reaction mixture was quickly added dropwise to deionized water (625 ml), and the temperature was maintained at 20-25. (3. The layers were separated, and the organic layer was washed with 500 ml of a 1 M NaHC03 solution. The organic layer was concentrated to separate (Gas carbonyl (third butoxycarbonyl) cyclopent-3-ene, pale yellow liquid. NMR (300 MHz, CDC13) δ 5.61 (s, 2H, CH = CH), 3.04 (app. Q, J = 15.1 Hz, 4H, 2CH2), 1.49 (s, 9H, C (CH3) 3). Han (film) 1796 (COC1), 1743 (C02R), 1274, 1233, 1158 cm-1. Tetrabutylammonium hydrogen sulfate ( 0.81 g, 2.4 mmol) in deionized water (700 ml), added to sodium azide (20. 16 g '3 10 mmol). It will contain 1 _ (chlorine 85435 -77- 200400815 Phenyl) -1- (third butoxycarbonyl) cyclopentane-3_diluted solution of MTBE / toluene 'was added to the azide solution for 45 minutes. The reaction mixture was stirred at 231: 3 hours until 1- ( Chlorocarbonyl) -1- (third butoxycarbonyl) ring Up to the consumption of pentamidine, it can be confirmed by GC analysis of the reaction solution. The layers were separated, and the organic layer was washed with 1 μ NaHCO3 (540 ml) and deionized water (540 ml, and then 270 ml). The organic layer was dried over NaJCU And filtered, and the solution was concentrated under vacuum to produce ι_ (fluorenyl azide) -1- (third butoxycarbonyl) cyclopent-3-en, which was oily. NMR (300 MHz, CDC13) δ 5.58 (s , 2Η, CH = CH), 2.96 (app t, J = 2.3 Hz, 4H, 2CH2), 1.46 (s, 9H, C (CH3) 3) IR (thin film) 2137 (CON3), 1720 (C〇2r ), 1246 (s, 1 185, 1136 cm-1 +). A solution of 1- (fluorenyl azide) -1- (third butoxycarbonyl) cyclopent-3-ene, added to 95 over 1 hour C in 110 ml of toluene. Under these conditions, the overflow of nitrogen is controlled by the adding speed. During the addition, MTBE is distilled from the reaction. The reaction is stirred at 95 ° C for 1 hour, and then allowed to cool to 23- Night. The solvent was concentrated under vacuum to produce 1_ (isocyanate) _ 丨 _ (third butoxycarbonyl) cyclopent-3- 晞, which was oily. LH NMR (300 MHz, CDC13) δ 5.67 (s, 2H, CH = CH), 3.01 j = 15.6 Hz, 2H), 2.61 (d, J = 15.6 Hz, 2H), 1.5 0 (s, 9H, C (CH3) 3) 〇 IR (thin film) 2258 (-NCO), 1732 (-C02R), 1157 cm -i. A second butanol (35 g, 471¾ mol) was added under nitrogen to a solution of the third butoxide (1M in THF, 471 ml, 471 mmol). The reaction solution was cooled to 0-5 ° C, and then a 1- (isocyanate) -1- (third butoxycarbonyl) cyclopentane_3-associated toluene solution was added, and the temperature was maintained at 0 ° -1 ° C over 60 minutes. . Add 85435 • 78- 2004 • 815 to 23 C, stir for 2 hours, and analyze the disappearance of isocyanate starting materials. The reaction mixture was added to a mixture of deionized water (12 liters) &amp; MTBE (12 liters) at 15 C. The solution was stirred for 20 minutes and the layers were separated. The organic layer was washed with 200/0 brine (250 ¾ liters) and the layers were separated. The organic layer was concentrated by distillation to about 250 ¾ liters. Heptane (500 ml) was added and the solution was concentrated to a total of 250 ml. The resulting slurry was cooled to 0 ° C, stirred for 2 hours and then filtered. The filter block was washed with cold heptane (2 x 100 ml), and then vacuum-dried to 34 54 g (52% yield, obtained from 1- (¾ ^ fe0-l- (second butoxycarbonyl) cyclopentane- 3_ 晞) of ι_ (third butoxycarbonylamino) -1- (third butoxycarbonyl) cyclopentane-3- 晞, as a white solid. Mp 87-89. (: LH NMR (500 MHz, CDC13) δ 5.63 (s, 2Η, CH = CH), 5.1 (bs, lR, NH), 2.99 (d, J-17.2 Hz, 2H), 2.57 (d, J = 16.0 Hz, 2H), 1.46 (s , 9H), 1.44 (s, 9H). 13C NMR (CDC13) 5 173.3, 154.9, 127.7, 81.1, 64.5, 44.8, 28.3, 27.8. IR (KBr): 3451, 2981, 2932, 1712, 1489, 1369, 1154 cm-1. MS (FIA) m / e (% relative strength) 284.2 (centi + 1, 56), 228.2 (73), 172.1 (97), 128.0 (100) 〇1- (third butoxycarbonyl ) -1- (Third butoxycarbonyl) cyclopentane_3_pyrene (200 g, 71 mmol) in 100 ml of EtOAc, added to tetrabutylammonium hydrogen sulfate (4.08 g, 10 mmol) 0.17 equivalent), magnesium monoperoxyphthalate hydrate (] ^ 1 ^? 8.8, 5 2.4 g '4.76% active oxygen = 2.1 equivalent) and deionized water (iqq ml) in a mixture. The reaction was stirred with a mechanical stirrer 19 hours. A solution of sodium hydrogen sulfate (18 g) in deionized water (100 liters) was added to the reaction mixture for 30 minutes. The mixture was diluted with 85435 -79- 200400815 100 ml of EtOAc and the layers were separated. The organic layer was deionized (100 ml), 2 N NaOH (2 × 100 ml) and deionized water (2 × 100 ml). The organic layer was distilled at 75-90 ° C and atmospheric pressure until the volume reached 40 ml. Solution Cool to 75 ° C, reheated heptane (120 ml). The solution is cooled to 65 ° C, and the desired product is implanted as needed. Let the resulting radon cool to ambient temperature, then cool in an ice bath for 1 hour The product was filtered, washed with 80 ml of cold 4.1 heptanol_EtOAc, and dried by suction at 40 ° C to give 15 45 g (73% yield) of the title compound 'as a white solid. Compound from 2 : 1 propanol: recrystallized from water for further purification. Mp 130-133 ° C. NMR (500 MHz, CDC13) δ 5.0 (bs, 1H), 3.58 (s, 2H), 2.41 (d, J = 15.3 Hz , 2H), 2.20 (d, J = 15.2 Hz, 2H), 1.43 (s, 9H), 1.40 (s, 9H). 13C NMR (CDC13) δ 171.3, 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, 840 cm ― 1. MS (FIA) m / e (% relative strength) 30.3 () ^ + 1, 65), 244.4 (68), 188.2 (100), 144.1 (99). Preparation of 46 (1 ba, 3¾. Bis (methyl fluorenylamine) propane dihydrochloride

85435

CH

Η

-80-

200400815 (R) -a-methylbenzylamine (98% ee, 121 g, 1 mole) was heated to nitrogen at 0 ° C under nitrogen. Dibromopromazine (25.4 ml, 505 g, 250 mmol) was added dropwise over 70 minutes. The mixture was heated for another 3 hours' and then cooled to 80 °. A concentrated (50%) NaOH solution (30 ml) was added dropwise over 10 minutes. Water (30 ml) was added to dissolve the solid particles' and the mixture was allowed to cool to room temperature for 30 minutes. MTT βE (100 ml) was added. Add 100 ml of water to dissolve the precipitate and separate the layers. The organic layer was washed with 50 ml of brine, dried (NajO4) and concentrated under vacuum to 1260 g of a pale yellow oil. The oil was distilled under a 70 t vacuum (head temperature) and a 12-inch tritium column was used to remove excess (R) -α-methylamidamine. The residue (68 grams of crude diamine) was dissolved in a 1 liter 1 to 1'011 in a 2 liter flask with a mechanical stirrer. Concentrated HC1 (12M, 45 mL, 540 mmol) was added dropwise over 10 minutes. Another 100 ml of i-prOH was added to ensure that the thick slurry was well mixed. The mixture was stirred for 90 minutes and then filtered. The filter block was washed with i_Pr0H, and then dried under vacuum at 40 ° C to obtain 64.32 g (72% yield, based on dibromopropane) of the title compound as a white solid. H NMR (500 MHz, DMSO-d6) δ 9.93 (bs, 2Η), 9.47 (bs, 2H), 7.56 (d, JT-7.0 Hz, 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 = 6.7 Hz, 6H) 〇 Preparation of 47 Ding Miaoxi cyclopentan-4-1-3-one 85435 • 81- 200400815 (1R, 3R) -1,3-bis (methylfluorenylamino) propane dihydrochloride (2465 g, 69.3 mol, Preparation 46) dissolved in 50 ml with stirring In the water. To the solution was added 35 ml (175 mmol) of a 5 N NaOH solution, followed by 150 ml of MTBE. After stirring for 15 minutes, the layers were separated, and the aqueous layer was extracted with 1 ml of MTβΕ. The combined organic layers were washed with 100 ml of brine, dried over Na 2 SO 4, and concentrated under vacuum to yield 18.06 g (92% yield) of ir, 3R_bis (methylamido) propylamine as a colorless oil shape. In a 500 ml four-necked flask (equipped with an overhead stirrer), dissolve 2059 g (73.5 mmol) of 1R, 3R-bis (methylamido) propane in 30 ml of anhydrous MTBE and THF ', And under & fluid. The solution was cooled to -45 ° C, and 59 ml (147 mmol) of n-BuLi solution (2.5 M in hexane) was added dropwise over 17 minutes. The pale yellow solution was stirred at -45 ° C for 2 hours. (LS, 3S, 5R) -3_ (third butoxycarbonylamino) -3- (third butoxycarbonyl) -6-fluorene bicyclo [3.1. 0] Hexane (10,000 g, 33.4 mmol), the temperature was kept below -40. (: Lower. Then rinse the addition funnel with 5 ml of THF. The reaction mixture was stirred at -45. (: Stirred for 18 hours, and then quenched by adding 4 N sulfuric acid aqueous solution (75 ml) dropwise. Once the acid was completely added The cooling bath can be removed. The layers are separated and the aqueous layer is extracted with 50 ml of MTBE. The combined organic layers are washed with 50 ml of water and brine, dried under Na2S04 and concentrated in vacuo to yield 9.78 g of crude 578242, white Solid. The solid was dissolved in 33 ml of hot MTBE and heptane (66 ml) was added in portions. Allow the stirred solution to cool to room temperature 'and stir the mixture at 0 ° C for 1 hour. Filter the solid to 2x10 Milled cold 2: 1 heptane: MTBE was washed, and then dried by suction at 35 ° C for 4 hours, and 8.52 g (85% yield) of (1S, 3R) -1_ (third butoxycarbonyl amine 85435) -82- 200400815 group) -1- (third butoxycarbonyl) _3_hydroxypent-4- 晞 'as a white solid. Mp 111-112T:. Analysis of palm HPLC: 99.7% ee [a] D = + 114 (c 1, MeOH) ° 'H NMR (500 MHz, CDC13) δ 6.1 (bs, 1H), 5.9 (bs, 1H), 5.55 (d, J = 5.0 Hz, 1H), 4.8 (m, 1H) , 4.44 (d, J = i〇.5 Hz , 1H), 2 87 (dd, J—14.5, 7.5 Hz, 1H), 2.00 (d, J = 14.5 Hz, 1H), 1 45 (s 9H), 1.42 (s, 9H). IR (KBr): 3413, 2983, 1703, 1491, 1370, 1309, 1255, 1155, 1055 cm-1. MS (FIA) m / e (% relative strength) 300.3 (M ++ 1, 15), 226.2 (29), 170.1 ( 100), 126.1 (89), 108.3 (20). Add 2,2,6,6-tetramethyl-1-hexahydropyridyloxy (radical) (teMPO) (0.84 g '5.3 mmol) Kiji 81 "(0.63 g, 5-3 millimoles in 2 ml of water) to (18,311) -1- (third butoxycarbonylamino) -1- (third butoxycarbonyl) -3-hydroxypentyl- A solution of 4-ene (20 g, 66.8 mmol) in 6 lb (200 ml). The reaction mixture was cooled to 0 ° C, and NaOCl (3.14%, 240 g) containing NaHC03 (8.4 g) was added dropwise. (100 mmol), the temperature was kept at 5 ° C. The reaction was stirred at 0 ° C for 1 hour, and allowed to warm to room temperature. The layers were separated and the aqueous layer was extracted with MTBE (2x200 mL). The combined organic layers were washed with 200 ml of 1 NHCi (containing 2.21 g of KI) followed by 10% Na2S03 solution (200 ml). The organic layer was washed with water (2x200 mL) and concentrated to dryness under vacuum. The crude product was dissolved in MTBE (60 ml) 5 (TC), and heptane (200 ml) was added for 1 hour to crystallize. The mixture was cooled to 0 ° C for 2 hours, and then filtered. 100 ml of cold hept 85435 • 83- 200400815 Calibration: MTBE (65.35) washing> strip filtration, block 're-vacuum drying to produce 16.99 g (89% yield) of the title compound as a white solid. Mp 116-18C 〇 [a] D = +123 (c 1, MeOH) NMR (500 MHz, CDC13) δ 7.4 (bs, 1H), 6.32 (d, J = 5.5 Hz, 1H), 5.6 (bs, 1H), 2.87 (d, J = 18.2 Hz, 1H), 2.9 (d, J = 18.2 Hz, 1H), 1.43 (s, 18H). 13C NMR (CDC13) δ 206.1, 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, 13〇s 1259, 1151, 1012 cm-1. MS (FIA) m / e (relative intensity./〇) 254.2 (M + + l, 11), 242.3 (18), 228.2 (13), 186.1 (76), 143.2 (11), 242.3 (100). Preparation 48: (2-tert-butoxy-2-oneethyl) ) IV i. Pyramidium 4 Potassium Bromide 0-C (CH3) 3 cd

Br 'tert-butyl bromoacetate (2.44 liters, 16.52 moles, 1 equivalent) was added to a solution of tetrahydropyridine (2.19 liters' 24.8 moles, 1.5 equivalents) in a solution of acetone (11.38 liters) at 22 The temperature of the flask was 30-60 minutes while maintaining the temperature at [5-25]. !: Between (tooth! Water bath). The reaction was allowed to fall for 22 hours, and the samples were analyzed by 1H NMR to verify that the reaction was complete. The precipitate was filtered, washed with acetone (2 liters), and aspirated and dried for 3 days at 28-33. (: The following compounds can be produced in 4.328 kg (92.5% yield 85435 -84- ratio). * H NMR (500 MHz, DMSOd-6) δ 4.40 (s, 1H), 3.51 (m, 2H) 3,48 ( m, 2H), 2.23 (m, 2H), 2.13 (m, 2H), 1.42 (s, 9H). Preparation 49 ΠS, 2S, 5R, 6R) -2- (Acedibutoxyzoylamine) _4 _ 嗣 Very-bicyclic "3.lo |. Hexafe -2,6-di-tert-acid di-tert-butyl ester

〖0 ^ 11 (42 ml, 42 millimoles, 1] ^ solution in Ding egg, 2.5 equivalents) Add soil 1_ (2-didioxy-2-S with ethyl) tetrakis Solution (11.9 g, 42 mM '2.5 eq, Preparation 48) in acetonitrile (30 mL) at 0 ° C solution, maintained at 10 ° C for 10 minutes. The milky solution was stirred for 1.5 hours on a cold calendar. Trifluoroethanol (6.9 g, 69 mmol, 4.1 equivalents) was added dropwise. Add (lS) -l- (Third-Butoxycarbonylamino)-(Third-Butoxycarbonyl) -Cyclopent-4-Fo-3-one (5 g, 16.8 mmol, Preparation 47 over 5 minutes) ) And trifluoroethanol (13.3 g, 132 mmoles' 7.9 equivalents) in acetonitrile (20 ml), the temperature was maintained between 3-5 ° C. The solution was stirred at 0-5 ° C for 4.5 hours. To the cold reaction mixture was added MTBE (155 ml) and H2O (80 ml). The layers were separated and the organic layer was washed with H20 (50 mL) and then with 20% brine (50 mL). The organic layer was concentrated to about 30 ml via an atmospheric pressure steam room. Heptane (100 mL) was added and the solution was concentrated. Add additional heptane if necessary until the vapor temperature of the distillate reaches 93 ° C. THF (65 ml) was added to form a solution of the title compound 85435 -85- 200400815. Preparation of 50 (lS, 2S, 5R, 6R) -2 dioxocarbonyl) _4_keto · bicyclo [3.] 〇ι Hexa-di-reportable acid di-tert-butyl ester 1.7 liters K / O3 saturated Aqueous solution was added to 1- (2-third-butoxy-2-ketoethyl) tetrahydrothiophene sulfide bromide (757 g, 2.67 moles, preparation 48) in 2.2 liters of CHsCh at 0 ° C, temperature Keep below 1 ° C. After stirring the two-phase mixture for 1.5 hours, add 223 ml of 50% NaOH solution in batches, and the temperature is kept below 5 ° C. The mixture is stirred for 3 hours and then filtered. The salt is rinsed with CH2C12. _ Lufen Layer, and the aqueous layer was extracted with 600 ml of CHiCb. The mixed organic layer was dried on a solid ruler 2:03 and concentrated under vacuum to yield 533.3 g (98% yield) of (2-third butoxy- 2-ketoethyl) tetrahydrop-phenphenamidine, a pale yellow oil. Once stored under refrigeration, the oil can crystallize to form a white solid. Mp 48-50 ° C. NMR (500 MHz, CDC13) δ 3.12 ( m, 2H), 3.00 (m, 2H), 2.85 (s, 1H), 2.41 (m, 2H), 1.84 (m, 2H), 1.38 (s, 9H). ▲ 474 ml (6.5 Moore 'l〇 (Equivalent) trifluoroethanol to (third butoxy condensed amino) -1- (third butoxy Base) _cyclopentane_4_ene_3 · ketone (194 g, 653. damr) in 650 ml of a solution of CH2C12 at 0 ° C. Added dropwise to 325 ml of CHWl2 (2-section A solution of tributoxy-2-ketoethyl) tetrahydro, thiophene (396 g, 1.96 moles), maintained at a temperature below 10 ° C. Remove the ice bath after 1 hour. Add deionized water (68 〇mL) and then separated. The aqueous layer was extracted with 400 mL of CH2Cl2. The combined organic layers were washed with 500 mL of brine, dried over Na2S04, and concentrated under vacuum to give 587 g of an amber oily solid. 85435 • 86- 200400815 Dissolved in 400 milliliter # CH2C12 'and dissolve it through a 1.6 kg silicone plug using 5: 1: 1 hexane: methyl tert-butyl ether: CH2C12 to form 13.2 liters of eluate. Concentration of the eluate can produce 398.7 g of white solid. The solid was dissolved in 3 liters of reflux: 30 hexanes: MTBE. The solution was allowed to cool to room temperature overnight and then cooled in an ice bath for 1 hour. The solid was filtered and rinsed with cold solvent (about 700 ml) ), And then dried under vacuum at 35 ° C to obtain 173 g (64% yield) of (lS, 2S, 5R, 6R) -2- (third butoxycarbonylamino) _4 _ Keto-Bicyclo [3.1.0] Hexane-2,6-dicarboxylic acid di-tert-butyl ester as a white solid. Mp. 144-46 ° C. [a] D = + 30.5 (c 1, CHC13). NMR (500 MHz, CDC13) δ 5.36 (d, 1H), 2.88 (m, 1H), 2.64 (dd, J = 5.2, 3.2 Hz, 1H), 2.37 (d, J = 2.7 Hz, 1H), 2.23 ( bs, 1H), 1.45 (s, 9H), 1.43 (s, 18H). 13C NMR (125 MHz, CDC13) 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 (CHC13): 2982, 1744, 1719, 1485, 1394, 1309 cm.1. MS (ES +) m / e (% relative strength) 412.2 (M ++ 1, 79), 356.2 (50), 300.1 (97), 276.1 (68), 244.1 (100). Analysis estimates C21H33N07 (411 · 29): C, 61.30; H, 8.08; N, 3.40. Found: C, 61.32; H, 8.04; N, 3.51. Preparation 51 Third butyl carbonyl amine amine) -4-hydroxy-bicyclo

H OH (ch3) 3c〇2c rY-C〇2C (CH3) 3 NHBoc The solution of preparation 49 was cooled to 0 ° C under nitrogen, and then tri-second butyl lithium borohydride (L-Selectride) was added dropwise. ™, 1 M in THF, 21 ml '21 mmol). The reaction mixture was stirred at 0 ° C for 4 5 minutes, and a 2M sodium bromide solution (31 liters) was added dropwise, and the temperature was maintained at 8 t :. A solution of 30% H2O2 (7.15 g, 63 mmol) in 20 ml of water was added, and the temperature was maintained at 15 ° C. After stirring for 10 minutes, add MTBE (210 ml) and deionized water (100 ml). The layers were separated and the organic layer was washed with a saturated Na2S03 solution (40 ml) and a 1 M NaHS04 solution (40 ml). The organic layer was concentrated to about 90 ml by distillation under atmospheric pressure. Continue to distill and add additional heptane to maintain a fixed volume. When the vapor pressure of the distillate reached 93 ° C, the solution was cooled to 70 t, and 7 ml of THF was added. The solution was cooled to 0 ° C and stirred for an additional hour. The solid was filtered, washed with cold heptane (10 ml), and dried under vacuum at 50 ° C to obtain 4.68 g (67% yield) of (13,28,43,511,6 feet) -2- (third butane Oxycarbonylamino) _4_hydroxy_bicyclo [3.1.0] hexane-2,6-dicarboxylic acid di-tert-butyl ester as a white solid. η.ρ. 187-88. . . ! H NMR (CDC13, 500 MHz) δ 5.31 (bs, 1H), 4.38 (d, JAB = 10.5 Hz, 1H), 4.30 (dd, J = 11.0, 6.0 Hz, 1H), 2.68 (d,) = \ 5 3 Hz 1H), 2.17 (m, 1H), 2.07 (m, 1H), 1.58 (m, J = 15.2 Hz 1H) 1.45 (s, 9H), 1.44 (s, 9H), 1.43 (s, 9H). 13C NMR (75 MHz, CDC13) δ 175.2, 170.5, 155, 83.2, 81 3 85435 -88-200400815 80.1, 73.7, 66.9, 43.1, 36.0, 34.4, 29.3, 28.3, 28.1, 27.4, 22.1 °

MS (ES +) m / e (% relative strength) 414.2 (M ++ 1, 58), 358.1 (75), 302.1 (78), 246.0 (100). (c 1, MeOH) 〇 Preparation of 52-yl-4_fluoro-dihydrofluorene 3.1.〇1 hexane_2.6-diazepine

(1 S, 2S, 4S, 5R, 6R) -2- (Third butoxycarbonylamino) _4_alkoxy_bicyclo [3 · 1 · 0] Hexan-2,6- A solution of Santin @ Mi (25.00 g, 60.5 mmol, Preparation 51) in CH2C12 (230 ml) was added to DeoxofluorTM (16.05 g '72 .6 Amor '1.2 equivalents) in ch2C12 (105 ml ) In -78 C solution for 2 hours. The reaction mixture was stirred for 15 hours, and 66 grams of Deoxofluor ™ was added. The solution was stirred for 30 minutes and allowed to warm to -⑺ ^. A saturated NaHC03 (105 ml) solution was added dropwise over 20 minutes, and the temperature was maintained at 5 ° C. The pH of the aqueous layer was adjusted to 7 by adding 160 ml of saturated NaHC: 03 solution. The mixture was allowed to warm to ambient temperature and then separated. The aqueous layer was extracted with 1 ml of ch2c12. The combined organic layers were washed with 250 ml of brine and dried over Na2SO4. The solvent 'was removed under vacuum and heptane (75 ml) was added to the crude product. 85435 -89- 200400815 The resulting mixture was heated to 50 ° C until all solid particles were dissolved, and then stirred at ambient temperature for 24 hours. The mixture was cooled to 0 ° C in an ice bath and filtered. The product was rinsed with cold heptane, and then dried under vacuum to produce 20.23 g (80% yield) of (111, 23, 4 feet, 511.6 feet) -2- (third butoxycarbonylamino) -4 -Fluoro-bicyclo [3.1.0] hexane-2,6-dicarboxylic acid di-tert-butyl ester as a white solid. m.p. 140-143. . . [a] D = + 20.6 (c 1, CHC13). ) H NMR (CDC13, 500 MHz) δ 5.45 (dd, JH-f = 56, Jh.h = 4.8 Hz, 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 (CDCI3, 75 MHz) δ 177.7, 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 strength) 416.2 (M ++ 1, 66), 360.1 (67), 304.1 (100), 248.0 (60). IR (CHC13): 3444, 2981, 1715, 1485, 1369 cm-1. S0C12 (21.29 g, 13.05 ml, 0.179 mole, 5 eq) was added to (lR, 2S, 4R, 5R, 6R) -2- (third butoxycarbonylamino) -4-fluoro-bicyclo [3.1. 0] Hexane-2,6-dicarboxylic acid di-tert-butyl ester (14.87 g, 0.036 mol) in a solution of EtOH (abs, 149 ml), added dropwise for 10 minutes, no cooling is required to generate a slight reflux Its solution. The solution was refluxed overnight. The solvent was removed from the reaction under vacuum. The residue was dissolved in EtOAc (150 mL), and a 10% Na2C03 (75 mL) solution was added, and added dropwise with stirring for 5-10 minutes. Layered to 85435 -90- 200400815

The aqueous layer was extracted with EtOAc (50 mL). The organic extract (1x50 ml) was washed with brine and dried over NazSO4. The product was filtered and concentrated under vacuum. (111,28,411,511,611) -2-amino-4-fluoro-bicyclo [3.1 .0] Hexane (-2,6-ditributyric acid di-tert-butyl ester is 'thick liquid' and can be solidified upon standing (1.12 g). N-methylmorpholine (22.44 ml, 204 mmol) to N-Boc-L-alanine (38.62 g '204 mmol) in a solution of 396 ml of dichloromethane at _22 ° C and nitrogen, followed by the dropwise addition of gas formic acid Isobutyl ester (26.48 ml, 204 mmol) over 15 minutes, so that the reaction temperature does not exceed _ 18. (:. Stir the resulting thin slurry at -20 ° C for 30 minutes, then add 247 ml (111,28,411,5 feet, 61 ^)-2-amino-4-fluoro-bicyclo [3.1.0] Hexane &gt; -2,6-dimetanoic acid diethyl ester (49.46 g , 191 millimoles) solution, the reaction temperature does not exceed -16 ° C over 40 minutes. Remove the reaction in the cooling bath and stir at ambient temperature for 70 minutes. Add 408 ml of 1 N hydrochloric acid, stir for 5 minutes, and then The layers were separated. The organic layer was washed with a saturated aqueous sodium bicarbonate solution. (1x408 ml), dried (Na2S04), filtered and concentrated, and (1R, 2s, 4R, 5R, 6R) -2- (2ιS-2l (third butoxycarbonylamino) propane) was obtained under vacuum. Group) amine group) _4 · fluoro-bicyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester as a white foam "&amp; 丨 6g). Add 46.7 ml (93.4 mmol) 2 N sodium hydroxide to crude (1R, 2S, 4R, 5R, 6R) -2- (2'S-2 '-(third butoxycarbonylamino) propanyl) aminofluoro_bicyclo [3 · 1.0] Diethyl hexane_2,6-dicarboxylic acid (7.5 g, 37.3 mmol) (theoretical value) in 46.6 ml of tetrahydrofuran in a solution at room temperature. The two-phase mixture was stirred vigorously at room temperature. Stir for another 1 hour (total 3 hours) until homogeneous. Dilute the mixture with 46 ml of tert-butyl methyl ether, mix for another 10 minutes and separate the layers. In separate flasks, add 93 ml of water, and then 84 ml ( 1〇1mmol 3〇3 85435 -91-200400815 ear) concentrated HC1. In the acid solution, add (1r, 2s, 4r, 5r, 6r) _2_ (2S 2 (di-butanylamino)) as needed. Propyl group) amine group), 4-fluorobicyclo [3 1 〇] hexane-2,6-dicarboxylic acid seeds, and Into the water layer from above. Add the water layer slowly to form a moderately thick slurry. At this time, increase the liquid addition rate (a total of 40 minutes of liquid addition time). Rinse with water (16 ml) Addition funnel. The resulting slurry was stirred for 2 hours, filtered, washed with water (2 × 32 ml), and filtered by suction at 45 ° C to constant weight, yielding 139 g (99%) of the title compound as a white solid. Preparation 53 a &amp; 4S’5S, 6S) -4z ^^ l ^ i ^ j isopropyl-2λ6- 喽 -difluorene di 1 ^ 6-dicarboxylic acid diethyl ester

Add thionyl dichloride to (1 &amp;, 48,58,63) -4-amino-2,2-diketo-2 into 6-ρ plug-bicyclo [3 · 1.0] hexane_4, A slurry of 6-dicarboxylic acid (10 g, 42.5 mmol, US Pat. No. 5,688,826) in 100 ml of 2B ethanol was added dropwise at room temperature (15.5 ¾ liter, 212.6 ¾ mol) After 20 minutes, rinse with 40 ml of ethanol. The slurry was heated to reflux and stirred overnight. The resulting solution was cooled to room temperature and concentrated to a gelatinous residue. To the residue was added ethyl acetate (ml), and it was diluted with another 94 ml of ethyl acetate.丨 5% sodium carbonate aqueous solution (70 ml) is slowly added to the mixture, and dissociation can be gradually generated under manual stirring to obtain a final pH value of 7.95. Filter and then layer. The aqueous layer was extracted with ethyl acetate (2X 1005 85435-92-200400815 liters). The combined organic extracts were washed with brine (1x100 ml), dried (MgSCU), filtered and concentrated under vacuum to give the title compound as a pale yellow oil, which solidified into a white solid (11.71 g, 95% yield). mp 80-83. . . [a] 25D-57.7 (c 1.04, CH3OH). 500 MHz lR NMR (CDC13) δ 4.31 (q, 2H, J = 7.0 Hz), 4.20 (m 2H), 3.78 (d, 1H, J = 15.0 Hz), 3.36 (dd, 1H, J = 4.〇, 7.0 Hz) 2.93 (dd, 1H, J = 4.0, 7.0 Hz), 2.81 (d, 1H, J = 15.〇Hz), 2.46 (t, 1H, J = 4.0), 1.34 (t, 3H, J = 7.0), 1.30 (t, 3H, J = 7_〇). 13C NMR (125 MHz, CD3C13) 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. Analysis estimates CnH17N06S: C, 45.35; H, 5.88; N, 4.81. Measured value: c, 45.02; Η, 5.75; N, 4.82 ° Preparation 54 (1 foot. 48,5 8.68) -4- (2'8-4 | -Methylsulfide-2,-(Third Ding 1 [ . 漶, amine, butan), amine-2.2-diketo-2, 6-far-dioxin, "3.1.01 inspection_ / 1 /;

〇S—CH. 85435 • 93- 200400815 N-methylmorpholine (14.4 ml, 130.9 mmol) was added to n-Boc-L-methionine (32.64 g, 130.9 mmol) at 11 〇 亳 liters of dichloromethane, in a clear solution under -22 ° C and nitrogen, 'followed by dropwise addition of isobutyl chloroformate (17 ml' 130.9 mmol) for 7 minutes to maintain the reaction temperature at -22 ° C. Once fully added, a thin slurry can form. Stir at -22 to -26 ° C for 30 minutes. Or, 4S, 5S, 6S) _4_amino-2,2-_keto_2_6-6-bis-bicyclo [3.1.0] hexanine in 107 ml of dichloromethane was added over 15 minutes- Diethyl 4,6-diallate (35.65 g '122.4 mmol, Preparation 53), followed by 36 ml of dichloromethane and rinsed. The reaction in the cooling bath was removed and stirred at room temperature for 70 minutes. 51 ml of 5N hydrochloric acid was added to the solution, and the layers were separated. The aqueous layer was re-extracted with two-gas methylamine (2xi07 mL). The combined organic layers were washed with a saturated aqueous sodium bicarbonate solution (107 ml), dried (MgSCU), filtered, and concentrated in vacuo to complete 65.82 g (103% weight yield) of the title compound as a white foam. [a] 25D -12.7 (c 1.2, CH3OH). 500 MHz NMR (CDC13) 6 7.53 (s, 1H), 5.06 (d, iH &gt; J = g 〇Hz), 4.34-4.20 (m, 6H), 3.41 (dd, 1H, J = 4.0, 7.〇) 5 2.97-2.89 (m, 2H), 2.64-2.59 (m, 2H, J = 4.0), 2.12-1.89 (m, 5H), 1.47 9H), 1.32 (t, .6H, J = 7.0). 13C NMR (125 MHz, CD3OD) 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. 85435 -94- 200400815 Preparation 55 oxycarbonyl ring) butyl ring Γ3 ″ · 01 P, dumplings_4 L -z 噼 monosodium salt 141 pens (282 耄 mol) 2 Ν sodium hydroxide is added to (1R, 4S, 5S, 6S) _4_ (2'S-4'-methylthio-2 '_ (third butoxycarbonyl) aminobutylfluorenyl) amino_2,2_diketo-2λ6-fluorene-bicyclo [3.1 .0] A solution of hexane-4,6-dicarboxylic acid diethyl ester (58 95 g, 112.8 * Moore, Preparation 54) in 141 ml of tetrahydrofuran at room temperature. The mixture was stirred vigorously at room temperature for 2 minutes. The solution was diluted with 141 ml of tert-butyl methyl ether and the layers were separated. The aqueous layer was further diluted with 141 ml of water and concentrated hydrochloric acid was added dropwise to lower the pH to 4.46. Get rid of it for 10 minutes to get a thin slurry. Adding more concentrated hydrochloric acid to the slurry lowered the pH to 1.4 (a total of 17 ml of concentrated HC1 '204 millimoles was used). After stirring for 2 hours, the slurry was filtered. The filter block was washed with water (2x118 ml) and then 45. (: Under vacuum drying, then transferred to a weighing pan. The filter block was dried again under vacuum at 45 ° C for 6 hours and 5 hours at 58t to produce 52.96 g (96% weight yield) of the title compound, white Solid. Mp (decomposition) 25 8 ° C. [&Lt; x] 25D -25,2 (c 1.03, H20). 500 MHz NMR (D20) δ 4.07-4.01 (m, 2H), 3.45-3 43 ( m 1H), 3.11 (d, 1H, J = 15.0 Hz), 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, CD3OD) δ 174.45, 173.58, 172.80, 157,46 85435 • 95- 200400815 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 56 (1R, 4S, 58, 68) -4- (23-4: methylthio-24 third butoxy , Amine, butanyl) amine-2,2-diketo-2λ6-fluorene-bicyclo 『3.1.01hexanox.-4 / -dipotent acid

397 ml (795 mmol) of 2 N sodium hydroxide was added to (lR, 4S, 5S, 6S) -4- (2'S-4'-methylthio-2 '-(third butoxycarbonyl) aminobutyridinyl ) Amine_2,2_diamidoyl-2λ-far-bicyclo [3.ι · 〇] Hexane-4,6-dicarboxylic acid diethyl ester (166.15 g, 318¾ mole, Preparation 54) Tetrahydroammonium 480 ml solution at room temperature. The mixture was stirred vigorously at room temperature for 2 minutes, at which time the reaction became homogeneous to form a clear pale yellow / green solution. Stir for another 2 hours at room temperature. The solution was diluted with 480 ¾ liters of &lt; fourth-butyl methyl ether and the aqueous layer was added dropwise to a solution of concentrated hydrochloric acid (71.5 ml '858 mmol) in water (960 ml). Ethyl acetate (500 liters) was added followed by the remaining aqueous layer to form an emulsion, which was further diluted with ethyl acetate (460 μl). Stir the emulsion for minutes, filter, and wash with water (2x tL). The filtrate was separated into layers and the aqueous layer was re-extracted with ethyl acetate (500 ml) 85435 -96- 200400815. The combined organic layer (75 mL) was washed with brine, dried (MgS04), filtered and concentrated under vacuum to yield 125.26 g (84% corrected yield) of the title compound as a white foam. Preparation 57 2- · / Smell-2-fluoro-2- (3-ketocyclofluorenyl) acetamidine g

8.45 ml (50.4 mmol) of triethylsilyl chloride was added to a suspension of 2.99 g of activated Zn (45.8 mmol) in anhydrous acetonitrile (100 ml) at -20 ° C. The mixture Stir for 5 minutes. 8.0 ml (57.3 mmol) of ethyl 2,2-dibromo-2-fluoroacetate was added, and the mixture was stirred at -201 for 90 minutes. Add 1.86 ml (22.9 mmol) of 2-cyclopentan-1-one, and stir the reaction mixture overnight to slowly raise the temperature to room temperature. 1 N HC1 (125 mL) and EtOAc (100 mL) were added. The organic layer was washed with saturated NaHC03 (2xl50ml), water (2xl50ml) and brine (2xl50ml), dried over anhydrous MgS04, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography using EtOAc / hexane (1: 8) as the solvent. The title compound (5.36 g, 88% overall yield) was produced as a diastereoisomeric mixture in the form of a colorless oil. . 'H-NMR (300 MHz, CDC13): 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).

Zn activation: Add 100 ml of concentrated HC1 to 100 g of zinc powder in a suspension of water (900 ml). The mixture was stirred at room temperature for 20 minutes. The water was decanted and the residue was washed with 97 · 3B9 8S43s 200400815 water (3x250 ml), acetone (3x150 ml) and ether (2x100 ml). The residue was dried overnight under reduced pressure at 35 ° C. Preparation 58 (lHS.5RS, 6RS) -6-i, -2-one bicyclo Γ3.1.01 Hexane-4,6- # i Acetyl Acetate

H F

Add 13.0 ml (75.0 mmol) of ethyl diisopropylamine to 2.0 g (7.5 mmol) of ethyl 2-bromo-2-fluoro-2- (3-ketocyclopentyl) acetate (Preparation 57) in DMF (8 ml) of the solution at 0 ° C, and the mixture was stirred at room temperature overnight. 1 N HC1 (20 mL), water (15 mL) and EtOAc (75 mL) were added. The organic layer was extracted, washed with saturated NaHCO3 (2 x 100 ml), water (100 ml) and brine (2 100 ml), dried over anhydrous MgS04, filtered and concentrated under reduced pressure. The residue was purified by column chromatography using EtOAc / hexane (1: 4) as the eluent to produce the title compound (1.17 g, 84% yield) as a mixture of isomers of trans: cis 5: 1, It was colorless oil. 'H NMR (300 MHz, CDC13): 1.33 (t, 3H, J = 7.1 Hz), 2.19-2.34 (m, 3H), 2.41-2.49 (m, 1H), 2.59 (d, 1H, J = 6.6 Hz ), 2.71-2.76 (m, 1H), 4.29 (q, 2H, J = 7.1 Hz). Preparation 59 (lRS, 2SR, _5RS, 6RS) -2-spiro-5’-Turbo Teng-6_f —The second ring "3.1.01 Jiyu &quot; 6- # m 85435 -98-

(lRS, 5RS, 6RS) 6-fluoro-2-one bicyclo [3 · 1.0] hexane-4,6-carboxylic acid ethyl alcohol (0.1 g, 0.54 mmol, Preparation 58) and 1 N NaOH ( A mixture of 0.55 ml, 0.55 mmol and EtOH (1 ml) was stirred under ice-cooling for 10 minutes. 1 N HC1 was added dropwise to the mixture until pH 1, and the resulting mixture was partitioned between EtOAc and brine. The aqueous layer was extracted twice with EtOAc, and the combined organic layers were dried over MgS04 and concentrated under reduced pressure. Residue, (NH4) 2CO3 (0.31 g, 3.2 mmol) and KCN (0.11 g, 1.62 mmol) in a mixture of EtOH and water (1: 1) (2 ml) at 60. (: Stir overnight. Cool in an ice bath and treat with 1 N KHSCU to acidify the mixture. Remove the solvent 'residue under reduced pressure, re-dissolve in MeOH, filter and concentrate in vacuo. The crude product can be used without further purification Use. Preparation of 60 D-6-fluoro-bicyclo "3_J.01 hexane-2,6-diphosphonate and π S.2R.5S-monoethyl-2- (21-third Amine-Propanyl-X-Fluoro-Dize 丨 1 1: Hexane-2,6-Diquinic Acid

Isomer A Isomer B 85435 -99- 200400815 L59 g (5.0 mmol) Ba (OH) 2 added to 0.32 g (1.5 mmol) (lR *, 2S *, 5R *, 6R *)- 2-spiro-5'-endourea · 6-fluoro-bicyclo [3J.0] hexane-6-carboxylic acid (Preparation 59) in a solution of water (10 ml), and The mixture was stirred overnight. Take 1 N HC1 at 0. (: Acidify the solution to pH 1 and concentrate under reduced pressure. The residue is redissolved and concentrated several times under reduced pressure with absolute ethanol until the solid is completely dry. 0.37 ml (5.0 mmol) ) SoCl2 was added to the residue in absolute ethanol (20 ml) at 0 ° C, and the mixture was refluxed for 5 hours. The solution was basified at a saturated value, and EtOAc (25 ml) was added. ). The organic layer was separated and the aqueous layer was extracted with more EtOAc (2x25 ml). The combined organic layers were dried over anhydrous MgS04, filtered and concentrated under reduced pressure. The residue was concentrated in 20 ml of DCM-DMF (4: 1), 0.73 g (1.9 mmol) HATU '〇_26 g (1.9 mmol) HOAt, 0.35 g (1.8 mmol) L-Ala mixture. Add 2.7 ml (15.3 mmol) Diisopropylethylamine and stir the mixture overnight under Ar. Add dcm (15 ml), separate the layers, and wash with saturated NaHC03 (2x25 ml), water (2x25 ml) and brine (2x25 ml). S. The organic layer was washed over anhydrous MgS04, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography using EtOAc / hexane (1: 2) As the eluent, the title compound (0.33 g '51% overall yield) was produced as a 1: 1 diastereoisomeric mixture as a colorless oil. The diastereoisomeric mixture was separated by palm HPLC The following analysis methods are used: 〇1such as 1-3] ^ 80 10 meters, 4.6 乂 250 mm; eluent: 10% 卩 in hexane; flow rate: 1.0 ml / min; 1; ¥ : 215 nm. Isomer A) Retention time = 5.9 minutes. Isomer B net retention time = 9.2 minutes. Isomer A: (1R, 2S, 5R, 6R, 2'R) -diethyl-2- (2'-tert-butoxycarboxyamine 85435 -100- 200400815 group-propanamido) -6- Fluoro-bicyclo [3.1.0] hexane-2,6-dicarboxylic acid NMR (300 MHz, CDC13): 1.18 (t, 3H, J = 7.1 Hz), 1.23 (t, 1H, J = 7.1 Hz ), 1.36 (s, 9H), 1.56-1.68 (m, 1H), 2.03-2.11 (m, 2H), 2.21-2.31 (ms 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 (75 MHz, CDC13): 13.9, 14.0, 17.7, 24.9 (d, J = 1.0 Hz), 2.81, 31.5 (d, J = 10.9 Hz), 34.9 (d, J = 9.4 Hz), 36.4 ( d, J = 8.8 Hz), 4.91, 61.5, 61.8, 67.4, 77.2, 81.5 (d, J = 242.6 Hz), 155.4, 168.6 (d, J = 25.1 Hz), 171.8 and 172.7 ppm. Isomer B: (1S, 2R, 5S, 6S, 2 | R) -diethyl-2- (2l-tert-butoxycarbonylamino-propanamido) -6-fluoro-bicyclo [3.1 .0] Hexane-2,6-dicarboxylic acid ester! H NMR (300 MHz, CDC13): 1.13 (t, 3H, J = 7.1 Hz), 1.18 (t, 1H, J = 7.1 Hz), 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? ιΗ)) 4.07-4.12 (m, 5H), 5.20 (sd, 1H), 7.22 (bs, 1H). nC-NMR (75 MHz, CDCI3): 13.7, 13.8, 18.0, 24.7, 28.0, 31.4 (d, J = 10.9 Hz), 34.5 (d, J = 8.9 Hz), 36.3 (d, J = 9.4 Hz), 49.1, 61.4, 61.7, 67.3, 77.2, 81.4 (d, J = 242.6 Hz), 155.2, 168.5 (d, J = 25.1Hz), 171.7 &amp; 172.8ppm. Preparation 61 [1R, 2S.5R, 6R) -2-Amino-6-gas-dihydrazone Π.1.01 hexane-2,6-dicarboxylic acid

85435 -101-200400815 30 mg (0.07 mmol) of a solution of 60 isomers in 6 N hci (2 ml) was refluxed overnight. The solvent was removed under reduced pressure, and the residue was washed with diethyl ether, dissolved in MeOh (1 ml), and propylene oxide (2 ml) was added. The mixture was stirred overnight at room temperature, the solvent was decanted, the residue was washed with ether and dried with Ar steam to give the title compound (12 mg, 85%) as a white solid. [ct] D = -25.0 (c = 0.80 mg / ml, h20). Preparation 62

Starting from the preparation of solution 60 of isomer B, the title compound can be prepared basically as in preparation 6. [ct] D = 21.7 (c = 0.46 mg / ml, H20). Preparation of 63 (place) -4-fluorene bicyclo [3 _ 1.01 hexane-2-iso-6-rollic acid ethyl ester

A three-necked 5 liter flask 'equipped with a stirrer, a thermocouple, and a Teflon tube, and an N2 inlet filled with 2000 ml of pyrimidine (d = 105 g / ml, starting Add 25-0 moles of diazoacetic acid ethyl acetate solution in the stilbene to increase to 45 moles to 17.1 equivalents, based on the total addition of diazoacetic acid ethyl acetate! Whichever is not corrected for EDA knowledge, if (According to the purpose of heavy gas acid g g-m 85435 -102. 200400815 correction strength is 19.0 equivalent). In this case, 0.968 grams of caprylic acid was added under Ns. 4 (1.24 millimoles' 0.0472 moles / °, based on the weight of ethyl diazoacetate without strength correction, and pure ethyl diazoacetate was used. Otometer is 0.052 mole%, Johnson Mathey: Lot No. 059255001). The suspension was heated to 46 ° C and stirred at 46 ° C for 10 minutes to affect the dissociation into a green solution. To this solution was added a solution of 300 g of diazoethyl acetate (90% purity, 2.63 moles, 2.37 moles pura '1.00 equivalent, Aldrich: Lot No. 17603PI) dissolved in 1600 ml of phenone via a drain pump. Such a liquid addition rate makes the total liquid addition time to 8 hours, and the slow liquid addition speed can prevent the formation of maleate and ethyl fumarate. When no diazoethyl acetate remained (about 30 minutes), this dark amber solution (3.985 g) was cooled to 23 ° C. The reaction mixture was divided into two parts, and the larger part (3.240 g of 3.985 g = 81.3% of the total weight) was directly concentrated to an oil. The crude product was degassed by a cloth-membrane distillation apparatus at 1.5 Torr and 23 ° C, and the remaining pyrimidine was removed. The product was then distilled at 1201: and 1.3 Torr. The title compound (light yellow) was collected in two portions: 155.0 g and 32.2 g. According to HPLC analysis, the strengths of these two parts were 78% and 76%, respectively. The crystals from the above distillate can be dissolved in T alcohol (2 ml per gram of distillate) and then cooled to -10 ° C. If no crystal growth is observed at this time, seed crystals can be seeded as needed. Once the crystals started to grow 'the mixture was further cooled to -45 C' and stirred for 2-3 hours, filtered, and washed with cold (-45.0) methanol (1 x 1 ml per 1 g of distillate). Substance at 24. (: Drying under vacuum to produce the title compound, white to white-doped solids' 80-85% recovery and &gt; 98% strength. Preparation of 64 G_S, 6S) -4-Hydroxy-2-2-4 tonne "3.1. 01 Hexane-6 · # ethyl acetate 85435 -103- 200400815

A solution of (6S) -4-pyrimadiazole [3.1.0] hex-2-ene-6-carboxylic acid ethyl ester (22.2 g, 131 mmol) in 136 ml of tetrahydrofuran under nitrogen at 0 ° C Add the borane-THF complex (98 ml, 98 mmol) over 15-20 minutes. After stirring at 0 ° C for 30 minutes, the reaction was allowed to warm to 15. (:, Stir again until it is complete by HPLC (1.5-2 hours). The reaction is cooled to 0. (:, transferred to 10 ml of pre-cooled 1 NpH7 buffer solution at 10-15 minutes, temperature Maintained at 0 ° C. To this mixture was added five parts of solid sodium perborate monohydrate (15.6 g '157 mmol), so that the temperature can be maintained at 20. (: next. Let the liquid drop add Warm to room temperature, stir for an additional hour and add 222 ml of water. After stirring for 2 hours, 'peroxide plus sodium thiosulfate pentahydrate (9.7 g) is dissolved in 24 ml of water and quenched'. The mixture was extracted with ethyl acetate (2 × 222 ml). The combined organic extracts were washed with a saturated aqueous sodium bicarbonate solution (1 × 222 ml), then with brine (1 × 222 ml) and concentrated to dryness under vacuum. The crude product is dissolved in 丨, 2_dichloroethane (each 丨 grams of crude oil 丨 milliliter) and then packed into a silica gel column (4 grams of silicone per gram of crude oil, slurried and packed in 15% acetic acid Ethyl-heptane). The column was dissolved with 15% ethyl acetate-heptane until it was visualized by TLC. At this point, the solvent was converted to 50% ethyl acetate-heptane acetate. All fractions containing the title product were mixed and concentrated under vacuum to an oil. The overall yield was in the range of 55-65%. Preparation 65 85435- 104- 200400815 L6S) -4- | g bis 1-fluorenyl bicyclic `` 3._1: 〇l £ u ^ -chitoate ethyl h, ch2co2c

To a solution of dimethylarsine (33.4 ml, 471 mmol) in 194 ml of dichloromethane in -7O ° C, slowly add trifluoroacetic anhydride (33.2 ml, 235 ¾ Mol) solution (temperature maintained at -66 ° C) over 30 minutes. After stirring for 20 minutes, (4S, 6S) -4-jiao-2-acylcyclo [3 丄 〇] hexane-6-ethyl acetate solution in 194 ml of dichloromethane was added over 60 minutes (341 Grams, 181 millimoles) 'so that the temperature was maintained at -60 () (: below. After stirring for 丨 hours, the reaction was treated with diethylamine (75.7 ml, 543 millimoles) for 35 minutes, so that the temperature was maintained at -5 (TC below. Let the reaction stir again! Hours, turn the cooling bath over time, and add 400 ¾ liters of 2N hydrochloric acid. Once warmed to 0t, the layers are separated and the organic layer is washed with 2N hydrochloric acid (1x300 ml), Followed by! Sodium bicarbonate aqueous solution (ΐχ67〇mL) and water (1x300mL), then dried over sodium sulfate, filtered, concentrated under vacuum ... folded into ... working color oil, once it can stand Cure. Crude product is added to a silicone tray. The starting alcohol is filled with 2 g of dichloromethane and dissolved in m (200-300 ml). All fractions containing the product are collected and concentrated to produce the title compound. Orange / brown oil. Typical calibrated yields are in the range of s5-90%. MM66 丄 01 hexane-4.5 drazolazole 85435 -105-

200400815 Ammonium carbonate (2.46 g, 25.6 millimoles, fen / Λ. Mao Guandou) and potassium iodide (0.87 mg, 12.5 house mole) were mixed in 19.9 ml of formic acid by human v T'in, and stirred. 30 minutes. The mixture was treated with a solution of (68) -4-keto-2-pyrimidobicyclo [31〇] hexane-6-carboxylic acid ethyl ester (2 39 g, .12.8 mmol) in 19.9 ml of methanol, and the reaction was heated. Stir to 30 ° C for another 23 hours. The volatiles were evaporated and the residue was dissolved in 2 75 ν sodium oxychloride (! 3. 丨 ml) and stirred for another 丨 hours. After diluting with 131 ml of water, the ρ 降至 value was reduced to 3.1 with concentrated hydrochloric acid, and then (6s, 11R) _8,1〇_Dione_2_ snail [—cyclo [3.1.0] hexane_4 , 5'_imidazolidine] 6-carboxylic acid β pH drops to 10, the suspension is allowed to cool to 0 ° C, and stirred for an additional 1.25 hours. The brown solid was collected, washed with cold water (2.3 liters and 0.8 ml), and then dried under vacuum at 400c overnight, which yielded 2.00 g (5 5% for purity correction) of (68,1111) _8,1 〇_diketone_2_thiaspiro [bicyclo [3.1.0] hexane_4,5, -imidazolidine] 6-carboxylic acid. The filtrate was diluted 'with 50 ml of ethyl acetate and treated with 18 g of sodium chloride. After stirring for 15 minutes, the layers were separated and the aqueous layer was further washed with ethyl acetate (2 x 50 ml). The combined organic extracts were dried over sodium sulfate, filtered, and concentrated under vacuum to a slurry (about 5 ml). To this was added a third butyl methyl ether (25 ml) and stirred overnight. The solid was collected, washed with tert-butyl methyl ether, and dried at 40X: for 2 hours under vacuum to yield 0.64 g (10% corrected for purity) of the title compound as a diastereomeric Sine vein 1.1 mixture. The two kinds of glandular gland are mixed with one kind before receiving the next step. 85435 -106- 200400815 a *. A slurry of racemic acid (15 g, 65.7 mmol, approximately 6: i diastereoisomeric hydantoin ratio) in 300 ml of ethanol and 75 ml of water Add (R) -phenylglycerol (9.0 g, 65.7 mmol). The mixture was heated to about 80 t to achieve dissociation. The dark solution was allowed to cool slowly, and precipitation was observed at 40-45r. The slurry was cooled further and held for 15 hours. The solids were collected and washed (with stirring) at 4: 1 ethanol: water (1x60 ml, pre-cooled to) at 65 ° C. (: Drying under vacuum for 12-24 hours. Typical yield of the resolved salt is in the range of 37-45%. 'Observable &gt; 98. And. &Gt; 98% ee. The resolved salt is soluble 6 times The volume (liters per liter) of water is treated with 1. 丨 equivalent of concentrated HC1. The slurry is cooled to 0 C, allowed to stir for 1 hour, then filtered, rinsed with 1 volume of cold water, and vacuumed at 60 ° C. Drying. The typical yield of the title compound is de% &gt; 90%-ee% &gt; 99% ° (MJ 18) -2,2,8,10-tetraketone-2-4 spiro "difluorene" 3.1. 01Hexane-4,5, -imidazolium 16- # 醯 Η 〇 \\ 々〇

〇 To a mixture of 6.8 ml of water '0.7 ml of 50% aqueous sodium hydroxide solution and 186 mg (0.74 mmol) of tungstic acid, add (6S, 11 S) -8,10-dione-2-pipispiro [· One $ clothing [3 · 1 · 〇] Hexan-4,5'-weitudian] -6-metanoic acid (3.4 g, 14.9¾ mole). The resulting solution was heated to 50 ° C, treated with 35% hydrogen peroxide (7.7 ml, 85435 -107- 200400815 74.5 millimoles), and slowly passed for 66 minutes. The reaction was allowed to stir, at 5 ° C for 5 hours, then cooled to 0 ° C, washed with thin Celite, and rinsed with cold water (1x2 ml). The filtrate was heated to 5 (rc, treated with concentrated hydrochloric acid to 1.5. The slurry was cooled to room temperature and stirred overnight. Once cooled to, filtered in the slurry 'washed in cold water (2x2 ml), and dried by suction at 55 ° C To constant weight, 3.19 g (82%) of the title compound was produced as a white solid. [A] 25D-48.6 (c, 1.19, 1 N NaOH). Mp 275 ° C (ash), 295T: (brown). 500 MHz lU NMR (DMSO-d6) δ 13.15 (br s, 1H), 10.99 (s, 1H), 8.13 (s, 1H), 3.85 (d, 1H, J = 15.0 Hz), 3.74 (dd, 1H, J = 7.〇, 4.0 Hz), 3.03 (d, 1H, J = 15.5 Hz), 2.80 (dd, 1H, J = 7.0, 4.0 Hz), 2.39 (t, 1H, J = 4.0 Hz). 13C NMR (125 MHz, DMSO-d6) δ 174.39, 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. Analyzed and estimated C8H8N206S: c, 36.93; H, 3_ 10; N, 10.77. Found: c, 36.76; H, 3.07; N, 10.60. Preparation of 68 Amine) -Propanamide 1 Amine-Γ2-fluorenyldi_3.1.01 Hexane &gt; 4,6-Di # 醢

85435 -108- In a stainless steel Parr reactor, (6S, 11 S) -2,2,8,10-tetraketone-2-thiaspiro [bicyclo [3 · 1.0 · hexane] -4,5 , -Imidazolidine] -6-carboxylic acid (2.50 g, 9.60 mmol) and 2 N sodium hydroxide (24.0 ml, 48.0 mmol). The mixture was heated to 95 ° C and stirred for another 21 hours. The mixture was cooled to room temperature and treated with activated carbon (125 g). The mixture was filtered through Celite 'and the filtrate was concentrated to 17 grams and then diluted with HsO to produce a weight of 24 grams. The pH was reduced to 6.5 using concentrated HC1 and the mixture was heated to 62 ° C. The pH value drops to 2.5 with concentrated HC1, and crystallization can occur. Allow the suspension to cool to 30 ° C, adjust the pH to 1.7, and lower the temperature. After the suspension was kept at this temperature for 18 hours, the solid was collected and washed with cold h20 (2x2, 9 ml). White solid at 45. Dry under vacuum to give the title compound (1.81 g, 80%). The title compound was slurried in 10 volumes of water and heated to 85 ° C for 3-4 hours, cooled to room temperature, stirred for 2-3 hours, filtered and washed with water (1 × 1 volume). Recovery rate> 95%. Preparation of 69 £ M1: _2-ketobicyclo `` 3.1.01 hexane-6-miao _ Λ ester

To a suspension of (ethoxycarbonylmethyl) dimethylsulfonated bromide (134 g, 585 mmol) in 486 ml of acetonitrile, 87.4 ml (585 mmol) of 1,8- Two pf bicyclic [5 · 4.0] 碳 carbon-7-woman, added dropwise over 15 minutes. After 1 hour of incubation, the yellow mixture was treated with 40 g (487 mmol) of 2-cyclopentamidine-1-埽 for 10 minutes. The mixture was left to stir overnight 'at this time, 480 ml of 85435-109-tributylmethyl ether' was added and washed with 1 N hydrochloric acid (1 x 24 0 liters). The aqueous layer was washed with second butyl methyl ether (1 × 240 ml). The combined organic extracts were washed with brine (1x400 mL), dried (MgS04) 'filtered and concentrated under vacuum to yield (6S) -2-onebicyclo [3 · 1 · 0] hexane_6 _Ethyl carboxylate as an orange solid (848 g). The crude product was purified by distillation (~ 138X :, 10 mm Hg), and the solidified steamed product was slurried in heptane, filtered and dried. Preparation of 70 (± 11 ^ 2-ketobicyclo Γ3.1.01)

A solution of crude (6S) -2-onebicyclo [3.1.0] hexane-6-carboxylic acid ethyl ester (30.2 g, 180¾ mol, uncorrected) in 30 ml of ethanol at room temperature Add 89 ml (178 mmol) of 2 N sodium hydroxide. After stirring for 80 minutes, the reaction mixture was washed with di-dibutyl methyl acid (1 × 90 ml), and the aqueous layer was treated with concentrated hydrochloric acid (18 ml) to reach pH = 1.0. The mixture was treated with 15 g of sodium chloride and washed with ethyl acetate (3 x 90 ml). The combined organic extracts were dried (Na2S04) 'filtered and concentrated under vacuum to yield 238 g (94%, uncorrected) of the title compound as a white solid. Preparation 7 1 (+) (place) -2-one bicyclic "3.1_〇1hexane- 6_Tae N- 芏 &amp; -α- 甲某 羊 fe 5¾ 85435 -110- 200400815

9 g of crude) S) -2-one bicyclo [3 丄 〇] Hexane_6 side acid, 84'9 millimoles, assuming 100% strength) in 119 liters 6: i ethyl acetate purpose: ethanol The solution was added under reflux to gram of molmol benzylamine. Once dissociated, allow the mixture to cool and seed as needed at 52 ^;:. -Once cooled to room temperature and stirred for an additional 13.5 hours, the crystals were collected and washed with ethyl acetate: ethanol (2 x 48 ml). Drying under vacuum produced 108 g (36% &apos; 77% de) of resolved salt as a solid.

The GC analysis of the derived methyl ester can determine the salt content, prepared as follows: 150 g of the resolved salt is dissolved in 5 ml of digas formamidine, and then washed with in sulfuric acid (2x1 liter). The organic layer was dried, filtered, diluted with 2 ml of methanol, and then 1¾ liters of 2 M monomethylsulphonyldiazomethane was burned in hexane. After stirring at room temperature for 15 to 4 li, the mixture was concentrated under vacuum to produce suitable gC: dialyzed methyl esters. GC conditions: 30 m. 25 mm x 25 μ p_DEX 325 "King 'HO C oven temperature, deuterium carrier gas just ml / min, FID detected at M. c, 1 microliter divided into 1: 100, Sample @ 1 mg / ml in dichloromethane. Preparation 72 ^^^^ 13.1. (Η oxane-6- # acid ethyl ester 383 85435 -111, Η

Suspension of 46.3 g (132 mmol) of (6S) -2-_bicyclo [3.lo] hexamidine-6-¾ acid N-fluorenyl-α-methylbenzylamine salt in 200 ml of ethyl acetate Add 198 ml (198 mol) of 2 N sodium hydroxide. After sufficient mixing, the layers were separated, and the aqueous layer was washed with ethyl acetate (1 × 200 mL). The aqueous layer was treated with 8 ml (2 丨 丨 moles) of concentrated hydrochloric acid and 100 g of sodium chloride. The mixture was allowed to stir for 30 minutes and then washed with ethyl acetate (2x200 ml). The mixed organic layer was dried (MgS04), filtered, and concentrated under vacuum to produce 18,3 g (99%) of the resolved acid [(+) (6S) -2-one-1% [3.1.0] hexane Burn_6-lean acid] was a white solid. Next, 10 g (71 mmol) of the crude analytical acid product was dissolved in 42 ml of ethanol and treated dropwise with 4 ml (71 mmol) of concentrated sulfuric acid. The mixture was heated to 45 C and allowed to stir for 75 minutes. Once cooled to room temperature, 42 ml of water and 20 ml of ethyl acetate were added, followed by 12 g of sodium bicarbonate. After stirring for several minutes, the mixture was washed with ethyl acetate (2 x 50 ml). The combined organic layers were dried (MgSCU) 'filtered and concentrated under vacuum to yield u g (92%) of crude (6 S) -2-one-cyclo [3.1 .〇] hexane-6-carboxylic acid ethyl ester, which was White solid. Crystallization from 6.1 / heptane: tertiary butyl fluorenyl ether (3.5 ml per gram of substrate) yielded the title compound in about 80% yield, and was determined by palm Gc analysis> 98% ee ° Preparation of 73 ethane gas carbonyl) bicyclic "3.1.01 hexa-2-associate 85435 -112- 200400815 Η

(6S) -2-ketobicyclo [3 · 1.0] hexane-6-carboxylic acid ethyl ester (380.1 g, 2.26 mol) and sulfuric acid (18 M, 6.3 ml, 0.11 mol) in isopropenyl acetate ( 2.26 liters) of the mixture was heated under reflux using a Dean-Stark apparatus for 2.5 hours, at which time GC analysis showed that the title compound was (6S) -2-onebicyclo [3.1.0] hexane-6-carboxylic acid ethyl ester 9: 1 Mixture. After removing 950 ml of solvent in 1 hour by distillation, GC showed a TF product / starting material ratio of 17.1. Additional isopropyl behenate (900 ml) and concentrated H2SO4 (3.15 ml) were added, and the mixture was stirred at reflux for another 15 hours, at which time GC showed a product / starter of 27: 1. After removing another 1.35 liters of solvent, the mixture was cooled to room temperature, and then diluted with MTBE (2 liters), ^ 0 (250 ml) and saturated aqueous NaHC03 solution (600 ml). The layers were separated and the organic layer was washed with brine (400 mL). The combined aqueous layer was extracted with mtbe (400 ml) and the combined organic layer was dried (NajO4), filtered, and concentrated to a dark red / brown oil (540 g). The crude oil was divided into two equal portions and filtered through a rapid SiO 2 filter (713 g in each batch) 'and dissolved at 10: 1 / heptane: ethyl acetate. Mixing the product-containing fractions from both was concentrated to give the title compound as a yellow oil (460 g, 97%; 90% corrected for solvent by NMR). Column chromatography on the gel was performed, and the product was dissolved in ethyl acetate / hexane (1: 5) to produce an analytically pure sample of the title compound as a colorless oil. [a] 25D +185 (c 1.48, CHC13). 500 MHz lH NMR (CDC13) δ 5.19-5.18 (m, 1H), 4.12 (q, 1H, 85435 -113- 200400815 J = 7.0 Hz), 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, 2.5 Hz), 1.25 (t, 3H, J = 7.0 Hz). 13C NMR (125 MHz, CDC13) δ 173.37, 169.01, 152.26, 111.56, 61.28, 32.47, 32.40, 29.72, 24.97, 21.67, 14.95. FTIR (CHC13) 3026 (m), 2985 (m), 1724 (s), 1272 (s), 1187 (s) cm-1. ES HRMS estimated CnHuNCUtM + NHUf 228.1236, found 228.1252 ° Preparation 74 (3 S, lR, 6R) -7-4-5-ketotricyclic Γ4 · 1 · 0.0 &lt; 2.4 &gt; 1Hengxuan ^ 3- # Brew

(6S) -6- (ethoxycarbonyl) bicyclo [3.1.0] hexan-2-fluoren-2-ylacetate (212.2 g '1.0mol) and 2,3-dichloro-5, 6-Dicyano-1,4-benzoquinone (252.0 g, Ml Mol) was heated to reflux in a 2.02 liter mixture of 1,4-di4alkane, and allowed to stir for 17 hours, at which time GC analysis showed complete conversion to (6S) -4-ketobicyclo [3.1.0] hex-2-ene-6-carboxylic acid ethyl ester. The mixture was cooled to room temperature and diluted with thf (564 ml). The mixture was cooled to 1,8-diazodicyclo [5.4.0] undec-7-association (377 ml, 2.52 moles) was added over 30 minutes, so that the reaction temperature was maintained below 10 ° C. The mixture was cooled again to 5 ° C, and the fourth butyl rhenium peroxide (70 wt% in water, 210 ml, 1 &gt; 51 mol) was added over 50 minutes, and the reaction temperature 85435 -114- 200400815 was maintained at 9 ° C or less. After the mixture was stirred for another 50 minutes, the reaction was filtered, and the brown filter cake was washed with MTBE (2 x 800 mL). To the filtrate was added 1.20 liters of 1 N HC1, and after thorough mixing, the layers were separated. The organic layer was washed sequentially with a saturated aqueous solution of NaHC03 (1.20 liters), a saturated aqueous solution of NaS203 (1.20 liters), and brine (600 liters). After the solution was dried (Na2S04), it was concentrated to an orange slurry, which was then diluted with 200 ml of heptane. The volatiles were evaporated to give an orange solid, which was triturated with 350 liters of heptane and filtered ', and the filter block was washed with additional heptane (2 x 75 ml). The collected solid was dried under vacuum at room temperature for 17 hours to yield 138.7 g (75%) of the title compound as a brown-yellow solid. Crystallization from butyl sulfide yielded an analytically pure sample of the title compound as a white solid. [a] d + 2.3 (c 1.20, CHC13), +8.40 (c 1.28, acetone); mp 129-130 ° C. 500 MHz! H NMR (CDC13) δ 4.16 (q, 2H, J = 7.0 Hz), 3.99 (t, 1H, J = 2.5 Hz), 3.24-3.23 (m, 1H), 2.96-2.94 (m, 1H) , 2.21-2.19 (m, 1H), 2.08 (t, 1H, J = 3.0 Hz), 1.26 (t, 3H, J = 7.0 Hz) 〇13C NMR (125 MHz, CDC13) 6 201.19, 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 ° Analysis and estimate C9H1 () 04: C, 59.34; H, 5.53. Found: C, 59.32; H, 5.43. Preparation of 75 (6S) -iLM Di; Red 3.1.01 Hexa-2- 埽 -6-oleate

85435 -115- 200400815 Η

Although the title compound is often used in situ for the preparation of (38,111,611) -7- ° -5-ketotricyclo [4.1.0.〇 &lt; 2,4 &gt;] heptane-3-carboxylic acid ethyl ester, it will contain The reaction mixture of this compound was filtered and the filtrate was evaporated to produce a colored solid, which gave an analytically pure sample of the title compound. The solid was suspended in ethyl acetate, the suspension was filtered, and the filtrate was concentrated-condensed. The residue was chromatographed on silica gel and dissolved in ethyl acetate / hexane (1: 5 to 1: 2) to give the title compound, which was recrystallized from hot ethyl acetate and chromatographed using the previous conditions to give the title compound. White solid. [a] 25D -268 (c 1.17, CHC13). mp 97-98 ° C. 500 MHz! H NMR (CDCIs) δ 7.60 (ddd, 1H, J = 5.5, 2.5, 0.75 Hz), 5.73 (dd, 1H, J = 5.0, 0.5 Hz), 4.15 (q, 2H, J = 7 〇Hz), 2.96-2.94 (m, 1H), 2.63-2.61 (m, 1H), 2.60 (t, 1H, J = 2.5 Hz), 1.26 (t, 3H, J = 7.0 Hz) 〇13C NMR ( 125 MHz, CDC13) 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. Analysis estimate C9H1Q03: C, 65 · 05; Η, 6.07. Found C, 64.97; H, 6.01 〇 Preparation 76 85435-116- 200400815 £ ^, 6ik_4_hydroxy-2-one bicyclic age Λ fermentation

(3S, 1R, 6R) -7-fluoren-5-one tricyclo [4 丄 0.0 &lt; 2,4 &gt;] Heptazone-3 ethyl leptate (36.3 g, 0.20 mole) in 667 ml of acetone The solution was stirred and treated sequentially with sodium acetate (36.1 g '0.44 mol)' sodium sulfide (65.8 g, 0.44 mol) and acetic acid (27-5 ml, 0.48 mol). The mixture was allowed to stir at 30 ° C for 15 hours, and the acetone was removed under vacuum to leave an ocher solid 'and then partitioned between ethyl acetate (323 ml) and H20 (323 ml). The layers were separated and the aqueous layer was washed with ethyl acetate (3x323 mL). The combined organic layers were washed sequentially with a saturated aqueous Na2s203 solution (364 ml) and a saturated aqueous NaHC03 solution (364 ml). Each aqueous washing was extracted with ethyl acetate (323 ml). The combined organic layers were dried (NajO4), filtered and concentrated to a red-brown oil, which was soluble in ml of ethanol. Evaporation of the volatile materials gave the title product as a reddish brown oil (418 g, 114%). Silica gel column chromatography was performed using ethyl acetate / hexane (1: 2 to 2: v). Crystallization from autothermal MTBE produced the analytically pure title compound as a white solid. [A] 25D +3.9 (c 1.39 , CHC13), +6.0. (C 1.69, MeoH). Mp 81-82 ° C. 500 MHz * H NMR (CDC13) δ 4.60 (br s, 1H), 4.16 (q, 2H5 J = 7.0 Hz), 2.66 (dd, 1H, J = 5.0, 4.0 Hz), 2.42-2,40 (m, 1H), 2.34 (dd, 1H, J = 19.0, 5.5 Hz), 2.24 (br d, 1H, J = 3.0 Hz) , 2.07 85435 • 117- 200400815 (d, 1H, J = 19.0 Hz), 1.91 (t, 1H, J = 3.0 Hz), 1.27 (t, 3H, J = 7.0 Hz) l3C NMR (125 MHz, CDC13) δ 209.74, 170.07, 69.04, 62.32, 43.47, 36.89, 34.95, 26.14, 14.83. FTIR (CHC13) 3607 (w), 3447 (w), 3025 (m), 2985 (w), 1739 (s), 1728 ( s), 1270 (s), 1187 (s) cm-1. Analytical estimate C9H1204: C, 58.69; H, 6.57. Measured value C, 58.48; Η, 6.63 ο Preparation of 77 phenyl ethyl) amino group l (2S, 4S, 6R) -2-Ammonia-4- and radical bicycloΓ3.1.01Hexan-6-Sour acid

Η PH

(4S, 6S) -4-Lightyl-2-one bicyclo [3.1.0] hexane-6-chitoate (68.2 g 'corrected to 60.0 g' 0.326 mol due to ethanol substitution) in ethanol (332 ml) and water (332¾ liters) were added (R) -methylethanamine (46.3 ml, 0.359 mol) and NaCN (20.8 g '0.424 mol)' temperature was maintained at 20 and 25. 〇between. Concentrated HC1 (35.3 liters, 0.424 moles) was added over 10 minutes and 4 minutes while maintaining the temperature above the reaction temperature. The dark brown mixture was stirred for one hour and then seeded with the title compound as needed to initiate crystallization. Suspension was stirred for 丨 hours. ′ The brown solid was collected. The air was passed through a filter and water (664 ml) was added. After the suspension was stirred for an additional 1.75 hours, the title compound 'was washed again with water (3 2 2 liters). / 85435 -118- 200400815

nr Wang / Dish "Small depths of sacrifice sacrifice • Sampling / thriller without disintegration [a] 25D +81.6 (c 1.18, CHC13). mp 70-72 ° C (decomposition). Hydrolysis of clear (wet weight 145 The title compound can be quickly separated without disintegration.

Hz), 1.64 (dd, 1H, J = 15.0, 5.0 Hz), 155_154 (m, 1H), 1.40-1.39 (m, 4H), 1.17 (t, 3H, J = 7.0 Hz) 〇 '13C NMR (125 MHz, CDC13) δ 170.54, 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), 1197 (m) cm-1. FAB HRMS estimates C18H23N203 [M + H] + 315.17〇9. Found 315.1704. Preparation 78 2- "((1110--1-benzyl 4_ 棊) amino group K2S, j ^ §jL) -4_ 衮 Certain bicyclic" 3 丄 oip.

Η PH

85435 -119- 200400815 p-[((1 foot) _1_phenylethyl) amino] (2S, 4S, 6R) _2_cyano_4_hydroxybicyclo [3.1.0] hexane-6 -A solution of ethyl carboxylate wet block (0326 mmoles in theory) in DMSO (220 ml), slowly add 30% h2O2 (44.5 ml, 0.226 moles). The temperature is maintained at 27 ° C the next. The temperature was reduced to 19t, and 5 N NaOH (52.3 ml, 0.262 mol) was added carefully and slowly over 15 minutes to maintain the temperature at 22 and 27. (: Between. An ice bath with a moderate capacity is required to handle the exothermic heat of this reaction. This brown heterogeneous mixture is stirred for 20 minutes at the above temperature range. HPLC shows that the starting material has been consumed to form the amidine intermediate. In the reaction another After stirring for 5 hours, Na2S03 (13.7 g, 0.109 mole) was added and the mixture was stirred for 15 minutes. At this time, the mixture was negative for peroxide using starch-kidney test paper. After adding 3 n NaOH (291 ml , 0.873 mole) 'The mixture was heated to 85 ° C and mixed for 18 hours. The homogeneous brown mixture was cooled to 3 (TC, concentrated HC1 was reduced to pH 3.6, while the temperature was maintained between 30 and 35 ° C. After the crystallization started at pH 3.6, the suspension was stirred for 15 minutes and then the pH was lowered to 2.5. After the mixture was stirred for another 10 minutes, it was cooled to 2 ° C 'and stirred for 2 hours, after which the gray solid was collected and then cooled to h2. 〇 (400〇mL) and EtOH (300mL) were washed. The collected solid was dried under vacuum at 45 ° C for 17 hours to produce 42.9g (from Preparation 18, 43%) of the title compound. In order to make the reaction in progress Generated titles can continue to be processed, but Recovered from the mother liquor in the following way. The ethanol part of the mother liquor is evaporated, and the residue is mixed with the aqueous part of the mother liquor. After H20 (485 ml) is distilled under reduced pressure, the pH of the mother liquor is 70 ml of 5 N NaOH and 5 ml of 50 % NaOH was adjusted to 12.9. After the solution was washed with n-Bilom (3 × 800 ml), its pH was adjusted to 2.5 with concentrated HC1, and the solution was concentrated. The residue was diluted with EtOH (100 ml) and the volatile matter was distilled 85435 -120- 200400815 hair (2X). The residue was diluted with EtOH (150 ml), and the brown solids each with an additional title compound and salt were washed with EtOH (75 ml). ) (: It can weigh 102 grams after drying under vacuum. The two title compounds are combined and used in the subsequent esterification [a] 25D +4.5 (c 1.41, 1 N NaOH). Mp 220 ° C (by Doped with white to gray), 28 (TC (brown). 500 ΜΗζ Ή NMR (D20, KOD) δ 7.39 (d, 2H, J = 7.0 Hz) 7 19- 7.04 (m, 5H), 3.92 (d, 1H, J = 5.0 Hz), 3.67 (q, m, j = Z7 '〇Hz), 1.76 (d, 1H, J = 15.0 Hz), 1.54-1.52 (m, m), 137 (dd m 1 = 15.0 , 5.0 Hz), 1.15 (d, 3H, J = 6.5 H z), l.l2 (dd, 1H J = 6 0 3.0 Hz), 0.92 (t, 1H, J = 3.3 Hz). 13C NMR (125 MHz, D20, KOD) δ 185.82, 182.96, 148 01 131.31, 129.97, 129.78, 74.99, 73.84, 58.78, 46.91 38 〇5 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), U88 cm_1 e Analysis estimates C16H19N05: c, 62.94; H, 6.27; N, 4.59. Found: c, 62.70; H, 6.21; N, 4.67. Preparation 79 2-"((1110-l-Ethylethylidene) amine" (2S., 4S, 6R.h ^ amino-4-A)-_ P. 1.01 hexane-6-carboxylic acid ethyl cool _ 85435 -121-200400815

Η〇ΗH3CH, C〇2C

Although the title compound is usually used in situ to prepare 2 2-[((lR) -l-phenethyl)

Consumption, the nitrile hydrolysis reaction mixture is partitioned between 'CH2C12 and h2O. After the organic layer was dried (MgS04) ', the residue was purified by silica gel column chromatography, and the title compound was formed using EtOAc / hexane (2: 1) to EtOAc as a white foam. [cx] 25D +61.3 (c 1.20, CHC13). 500 MHz 4 NMR (CDC13) δ 7.32-7.20 (m, 5H), 7.19 (br d, 1 J = 4.0 Hz), 5.49 (br d, 1H, J = 4.0 Hz), 4.88 (d, 1H , J = ii.5 H2), 4.24 (dd, 1H, J = ll.5, 6.0 Hz), 4.06-4.00 (m, 2H), 3 77 (q, lH, J = 7.0 Hz), 2.21 (d , 1H, J = 15.0 Hz), 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 · 5

Hz), 1.32 (t, 1H, J = 3.3 Hz), 1.24 (t, 3H, J = 7.0 Hz). 13C NMR (125 MHz, CDC13) 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 ° 85435 -122- FTIR (CHC13 ) 3441 (m), 3345 (m), 2975 (w), 1725 (s), 1665 (s), 1288, 1186 (m) cm-1. Analysis estimates C18H24N204: C, 65.04; H, 7.28; N, 8.43. Measured value: c, 65.41; H, 7.58; N, 8.32 ° Preparation of 80 idiil RV phenethyl E) amine K2S.4S, 6R) -2- (Ye. Miao) _4 · 衮 环 「3.1.0101 Alkan-6- # acid ethyl ester

P-[((lR) -l-benzylethyl) amino] (28,48,6 p-4-hydroxybicyclo [3.1.0] hexane-2,6-dicarboxylic acid (4 g (13 mmol) in a 48 ml ethanol suspension. 'Ethyl chloro (U. 2 ml, 157 mmol) was added at room temperature, and a gentle reflux was maintained through the addition funnel. The mixture was stirred under reflux for another 16 hours, and once cooled to room temperature, it could be concentrated under vacuum to a solid residue. The solid was slowly treated with a solution of sodium bicarbonate (6.6 g) in 100 ml of water, and then Ethyl acetate is used to make ice, and 欲, v 彳 Λ ΛΛi, _., Θ λ ,,, ...

The title compound 'was a white solid. [a] 25D +52.5 (c 1.30, CHC13) = mp 73-74 ° C. 85435 -123200400815 500 MHz 4 NMR (CDC13) δ 7.29-7.14 (m, 5H), 4.25 (dq, 1H, 11.0, 7.0 Hz), 4.18 (dd, 1H, J = 9.5, 5.5 Hz), 4.10 ( dq, 1H, J = 11.0, 7.0 Hz), 3.92 (dq, 1H, = 11.0, 7.0 Hz), 3.82 (dq, 1H, J = 11.0 Hz, 7.0 Hz), 3.67 (q, 1H, J = 7.0 Hz ), 2.73 (d, 1H, J = 9.5 Hz), 2.15-2.12 (m, 2H), 2.01-1.99 (m, 1H), 1.89 (dd, 1H, J = 6.0 3.0 Hz), 1.61 (dd, 1H , J = 15.0, 6.0 Hz), 1.36 (t, 1H, J = 3.5 Hz), 1.33-1.30 (m, 6H), 1.18 (t, 3H, J = 7.0 Hz). 13C NMR (125 MHz, CDC13) δ 178.11, 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. Analysis estimates C20H27N05: C, 66.46; H, 7.52; N, 3.88. Found: c, 66.42; H, 7.44; N, 3.92. Preparation 81 2-I ((lR): JL · phenyl) amino group K2S, 4R, 6R) -2- (ethane hydrazone) -4-gas bicyclo Π. 1.01 hexane-6- # _ ethyl ester

P-[((lR) -1-phenylethyl) amino] (2s, 4S, 6R) -2- (ethoxycarbonyl) -4-hydroxybicyclo [3.1.0] hexane_6_ Ethyl carboxylate (59.0 g crude, 0.163 moles) in CH2C12 (690 ml). Deoxo-Fluor® 85435 -124. 200400815 (45.1 ml, 0.245 moles) was added at -20 ° C. Calendar The temperature was maintained between -15 and -20 ° C for 15 minutes. The mixture was stirred at this temperature for 20 minutes' and at 0 ° C for 15 minutes, after which a 15% aqueous Na2CO3 solution (650 ml) was slowly added, and the temperature was maintained below 10 ° C. The layers were separated and the aqueous layer was extracted again with CH2C12 (150 mL). The combined organic layers were dried (Na2SCU) and concentrated to a brown oil (73 g). This oil was purified on a silicone pan (400 g) and dissolved with EtOAc / heptane (1: 6) to give the title compound as a yellow oil (49.7 g, 84%). [a] 25D +36.2 (c 1.30, CHC13). 500 MHz ιΗ NMR (CDC13) δ 7.29-7.14 (m, 5H), 5.22 (ddt, 1H, ® J = 8.0, 4.5 Hz, Jhf = 56.0 Hz), 4.16 (dq, 1H, J = 11.0, 7.0 Hz) , 4.05 (dq, 1H, 11.0, 7.0 Hz), 3.96 (dq, 1H, 10.5, 7.0 Hz), 3.85 (dq, 10.5, 7.0 Hz), 3.66 (q, 1H, 6.5 Hz), 2.45 (dd, 1H , J = 14.〇, 8.0 Hz), 2.16-2.12 (m, 1H), 1.95 (t, 1H, J = 3.5 Hz), 1.81 (dt, 1H, J = 3.5 Hz, JHf = 3.5 Hz), 1.51 (ddd, 1H, J = 14.0, 8.0 Hz, Jhf = 22.0 Hz), 1.32 (d, 3H, J = 6.5 Hz), 1.27 (t, 3H, J = 7.0 Hz), 1.21 (t, 3H, J = 7.0 Hz). 13C NMR (125 MHz, CDC13) δ 175.29, 171.66, 146.21, 128.45, 127.03, 126.90, 92.65 (d, JCf = 182 Hz), 68.68 (d, JCF = 4.9 Hz), 61.70, 60.92, 56.13, 38.60 ( d, JCf = 23.0 Hz), 33.07 (d, JCF = 7.6 Hz), 32.23 (d, JCf = 22.0 Hz), 26.26, 20.22 (d, JCf = 3.9 Hz), 14.41, 14.24. FTIR (CHC13) 3028 (w), 2983 (w), 1724 (s), 1705 (s), 1293 (m), 1242 (m), 1190 (m), 1037 (m), 1013 (m) cm- 1. Analysis estimates C20H26FN04: C, 66.10; H, 7.21; N, 3.85. Measured value: C,

85435 -125- 200400815 66.〇2j Η, 7.00; N, 3.95 ° Preparation 8?

H F h.ch2co2

CO, CHXH 2-[((lR) -l-phenylethyl) amino] (2s, 4R, 6R) -2- (ethoxycarbonyl) _4_fluorobicyclo [3.1.0] hexane-6- Ethyl carboxylate (68'4 g, 0.188 mol), concentrated 11 (1: 1 (15.7 ml '0.188 mol)) and 10% pd / C (dry, 13.7 g) KEtOH (400 ml) The mixture was placed under hydrogen (50 pSi) for 18 hours. The catalyst was filtered off and the filtrate was evaporated to give the title compound as a white foam (59.2 g, 106% corrected to 97% due to EtOH doping). From EtOAc / Crystallization in MTBE yielded an analytically pure sample as the title compound as a white solid. [A] 25D +55.6 (c 1.17, CHC13). Mp 86-88 ° C. 500 MHz * H NMR (CDC13) δ 9.20 (br s, 2H), 5.50 (ddt, 1H, J = 8.0, 4.5 Hz, JHF = 56.0 Hz), 4.31 (q, 1H, J = 7.0 Hz), 4.20-4.07 (m, 3H), 2.88 (t, 1H , J = 3.0 Hz), 2.71 (dd, 1H, J-14.5, 8.0 Hz), 2.48-2.43 (m, 2H), 2.16 (ddd, lH, J = 14.5, 7.5 Hz, JHf = 22.0 Hz), 1.34 (t, 3H, J = 7.0 Hz), 1.25 (t, 3H, J = 7.0 Hz). 13C NMR (125 MHz, CDC13) 6 171.12, 169.41, 91.94 (d, Jcf = 189 Hz), 63.85, 63.66 ( d, Jcf = 3.8 Hz), 61.73, 34.55 (d, 85435 -126- 2004008 15

Jcf = 26.4 Hz), 31.58 (d, JCF = 7.8 Hz), 30.80 (d, JCf = 24.1 Hz), 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. Analytical estimates (: 121118? 1 ^ 04: c, 48.74; Η, 6.48; N, 4.74. Found: c, 48.80; Η, 6.41; N, 4.76. Preparation of 83-4-f, the second ring "3.1 〇1 hexane-2.6-two-volume age

3 N NaOH (251 ml. 〇.753 mole) solution was slowly added to 1R, 2S, 4R, 5R, 6R-2-amino-cardiofluorobicyclo [3_ 1.0] hexane-2,6-dicarboxylate Hydrochloride (59.2 g, crude, 0.188 mole theory), temperature maintained at 26 ° C. After the mixture was stirred for 10 minutes, it was homogeneous. The mixture was stirred at room temperature for 1.25 hours' and the pH was slowly reduced to pH 2.8 with concentrated HC1 while maintaining the temperature between 20 and 26 ° C. At pH 2.8, the mixture started to crystallize, and the suspension was stirred at this pH for 10 minutes, after which the pH was reduced to 2.1 with concentrated HC1. After stirring for another 15 minutes, i-PrOH (67 ml) was added and the suspension was cooled to 0 ° C and stirred for another 2 hours. The solid was collected and washed with 37 ml of cold H20 / i_POH (4: 1). The collected solid was dried under vacuum at 40 ° C for 18 hours to give the title compound as a white solid (33.1 g, 87% from the beginning of Preparation 23). Preparation 84

85435 -127- 200400815 1 ^ 28,411.511.6 feet-2-amino-4-fluorobicyclo "3.1 ^] Hex: burning -2,6-^^^^ Further to the 1st Han, 28,411,511,611- Suspended suspension of 2-amino-4-fluorobicyclo [3,1.〇] hexane-2,6-diamino acid (33.0 g, 0.162 mmol) in H20 (165 ml) Warm to 89 ° C for 1 hour, then add i-PrOH (41 mL). Stir the mixture under reflux (83 ° C) for 5 minutes, then allow it to cool to room temperature and stir for 4 hours. Collect the product to Wash with i-Pr0H / H20 (1, 4, 40 ml), then with i-prOH (25 ml), and dry under 401: vacuum for 18 hours to give the title compound as a white solid (30.6 g, 93%) Preparation of 85 111, 28.4 only. &Lt; 1 ^ 611-2-amine a certain 4-tired. Dioxane "3.1.01 hexane-24-dicarboxylic acid ethyl

£ ^ CO2CH0CH NH. For 1 foot, 23,411,5 feet, 611-2-amino-4-fluorobicyclo [3.1.0] hexane-2,6-dicarboxylic acid (14.45 g '71.12 mmol) In a slurry of 202 ml absolute ethanol at room temperature. Add thionyl chloride (26 mL, 356 mmol) dropwise over 20 minutes. The slurry was heated to reflux * 'and allowed to stir for 3 hours and then cooled to room temperature overnight. The resulting solution was concentrated to a residue under vacuum. It was diluted with 136 ml of ethyl acetate, and then treated with 306 ml of a 10% sodium carbonate aqueous solution for 15 minutes, and manually decanted to an initial pH of 10 °. The layers were organic and organic. The extract was washed with ethyl acetate (1 x 136 ml). The combined organic extracts were washed with brine (1 x 136 mL), dried 85435-128-200400815 (MgS04), filtered and concentrated under vacuum to yield 17.07 g (93%) of the title compound as a white solid. FDMS: M ++ l = 260. Analysis estimates C12H18FN04 * 0.1 H20: C, 55.21; H, 7.03; N, 5.37. Found: C, 55.10; H, 6.96; N, 5-22. m.p. = 64-66 ° C 0 [a] D25 = + 20 (c = 0.96, MeOH), [a] D25 = + 15 (c = 1,21, DMSO). M Preparation 86 iEJSjRJRjm-U’SU The third butoxygen and some amines, and some amines, 1-4, Qi 2nd board "3.1.01 Jiyu"-2,6-ethyl acetate

Η ΗΝ ^ Ο HX NHBoc

X To N-Boc-L-alanine (38.62 g, 204 mmol) in 396 ml of dichloromethane at -22 ° C under argon, add N-methylmorpholine (22.44 ml) , 204 mmol), and then isobutyl chloroformate (26.48 ml, 204 mmol) was added dropwise over 15 minutes, so that the reaction temperature does not exceed -18 ° C. The resulting thin slurry was stirred at -20 ° C for 30 minutes, at which time 1R, 2S, 4R, 5R, 6R-2-amino-4-fluorobicyclo [3.1.〇] hexane-2 was added over 40 minutes. A solution of ethyl 6,6-dicarboxylate (49.46 g, 191 mmol) in 247 ml of dichloromethane, so that the reaction temperature does not exceed -16 ° C. Once the addition is complete, the reaction is removed from the cooling bath and allowed to stir at ambient temperature for 70 minutes. At this time, the reaction temperature has shown 85435 -129- 200400400815 to 15 ° C, and the color It turns light orange. The reaction was treated with 1 N hydrochloric acid at 408 mmol and was removed for another 5 minutes and the layers were separated. The organic layer was washed with a saturated &lt; 5 anti-nitrogen steel solution (1 x 408 ml), dried (Na2s04), filtered 'and concentrated in vacuo to a white foam (88.16 g). FDMS: M ++ l = 260. Analyze CuHuFNCVoj h20: C, 55.21; H, 7.03; N, 5.37. Found: C, 55.10; H, 6.96; N, 5.22. = C. [a] D25 = + 20 (c = 0.96, MeOH), [a] V5 = + 15 (c = 1.21, DMSO). Preparation of 87 窠 Tributane Gong amine） propyl 醯 醯 1 amine ring [3.1.01 ^ alkane-2.6-dicarboxylic acid

H 〇2 c C〇2H Η ΝΝ

X 0 NHBoc 1-foot, 23,411,511,6 Han-2- [2'3-2,-(Third-butoxycarbonylamino) propanyl] phenyl-4-fluorobicyclo [3.ι_ 〇] Hexane_2,6-dicarboxylic acid ethyl ester (8816 g '191 mmol) in 238 ml of tetrahydrofuran, and at room temperature, 238 ml (77 moles of μN hydroxide surface was added. Let two The phase mixture was stirred vigorously at room temperature for 2.5 hours. At this time, the reaction was homogeneous. The mixture was diluted with 238 ml of tert-butyl methyl ether and then mixed and separated. The aqueous layer was further diluted with 23.8 ml of water and then washed. The particles were removed. The solution was treated with concentrated solution (ml, 515 mmol) for 30 minutes, followed by seeding with the title compound and stirring for 1 hour. 85435 • 130- The resulting slurry was filtered, washed with water (2x100 ml), and dried by suction at 45 ° C for 40 hours to yield 72.2 g of the title compound as a white solid. A portion of the solid (69.5 g) was stirred with 490 ml of acetone for 1 hour 'and the resulting hazy solution was filtered and washed with acetone (2 x 100 ml). The filtrate was concentrated in vacuo to a white foam at 45 ° C. (: Further drying under vacuum for 16 hours can produce 61.8 g (corrected for 12% wt / wt acetone) of the title compound. Example 1 (111, 48.58, 68) -4- (2 | 8- 眛 Very Bing ) Amino 1-2,2-diketo-2 human 6-1? 1-difluorene "3.1.01 Hexane-4,6-dioxoate

H3c ^ NH. P- (lR, 4S, 5S, 6S) -4- (2'S-third butoxycarbonylamino-propionamido) _ 2,2-diketo-2λ6-pyrimidine-bicyclo [3 · 1 · 〇] hexane_4,6_ dicarboxylic acid (1100 g, 271¾ mole, Preparation 3) in a suspension of 563 ml of ethyl acetate, and hydrogen chloride was added to ethyl phthalophthalate solution (3.7 M , 5 14 ml) over 20 minutes. Suspension agitation

The title compound (85.77 g, 92%).

85435 -131-200400815 1H, J = 4.1 Hz), 1.52 (d, 3H, J = 7 ″ Hz). Example 2 Anji _ Aceyl) Amine-2. End-4,6- 二 # 醢 甲 幕 碏 酩

(lR, 4S, 5S, 6S) -4- (2'S-third butoxycarbonylamino-propanamido) _2,2-monoketo-2 into 6-fluorenyl-bicyclo [3.1.〇] hexane Suspension of _4,6_dicarboxylic acid (300 mg, 0738 millimoles' Preparation 3) and toluenesulfonic acid monohydrate (14 mg, 0.738 millimoles) in toluene (3 mL) Liquid, heat to it and stir for 45 minutes, then let it cool to room temperature and stir for 16 hours. The suspension was filtered and the filter cake was washed with toluene (2 x 1 mL). After 1 hour of vacuum drying at 45 ° C, 307 mg (87%) of the title compound was collected as a white solid. mp (DSC) 233 ° C. lH NMR (500 MHz, CD3OD) δ 7.70 (d, 2H, J = 8.5 Hz), 7.24 (d, 2H, J = 8.0 Hz), 4.11 (d, 1H, J = 15 Hz), 3.94 (q, 1H , J = 7.0 Hz), 3.53 (dd, 1H, J = 7.〇, 4.0 Hz), 3.13 (dd, 1H, J = 14, l.〇Hz), 3.02 (dd, 1H, J = 7.0, 4.5 Hz), 2.48 (t, 1H, J-4.5 Hz), 2.37 (s, 3H), 1.52 (d, 3H, J = 7.5 Hz). 13C NMR (125 MHz, CD3OD) δ 170.70, 170.32, 169.80, 142.04, 140.78, 128.79, 125.79, 60.20, 54.73, 48.77, 42.44, 30.84, ifi 4 85435 -132- 200400815 22.22, 20.20, 16.09 ° Example 3 nR, 4S, 5S, 6S) -4- (2S-amine-3, hydroxy-1-propanylamino-2,2-diketo-2λ6-di-bicyclo "3.1.01 hexane-4 , 6-dicarboxylic acid hydrochloride

Prepared according to the general procedure C, using (1R, 4S, 5S, 6S) -4- (3'_acetamido-2'S-third butoxycarbonylamino-propanyl) -amino-2,2- Diketo-216-fluorene-bicyclo [3.1.0] hexane-4,6-dicarboxylic acid dimethyl ester (380 mg, 0.77 mmol, Preparation 15). [4 = -19.23 (c = 0.52, MeOH). 'Η NMR (400 MHz, CD3OD) δ 2.48 (1H, t, J = 4.0 Hz), 3.04 (1H, dd, J = 4.4, 7.3 Hz), 3.17 (1H, d, J = 14.7 Hz), 3.51 ( 1H, dd, J = 4.4, 7.0 Hz), 3.77 (1H, dd, J = 7.0, 10.6 Hz), 3.94-4.12 (3H, m). Analytical estimates (^ .1114 &gt; ^ 088 * 11 (: 1 *% 0 ·· C, 31 · 88; H, 4.55; N, 7.44; Cl, 9.41. Found: C, 31.53; H, 4.40; N, 7.32; Cl, 9.24. HRMS estimated C10H15N2O8S, 323.0549. Found 323.0533. Example 4 (111,48,5 8,68) -4-"(Pyrrolidine-23-carbonyl) -amine-1.2.2-dione -216-4 -Bicyclic "3.1.01 Hexane-4,6-dicarboxylic acid hydrochloride 85435 -133-200400815

Prepared according to general step C, using (1R, 4S, 5S, 6S) -4-[(r-third butoxycarbonyl p ratio HYD-2 S-carboxyl) _amino] -2,2-dione- 2λ6-ϊ »Sep-bicyclo [3 · 1 · 〇] hexanidine 4'6 acid-formamidine (0.8 g, 1.7 mmol, preparation n) yielded 0.42 g (67.0%) of the title compound. [«] 〇 = -32 (c = 1.〇〇5 CH30H) 〇'H NMR (300 MHZj CD3〇d) δΐ. 98-2.12 (3H, m), 2.40-2.50 (1H, m), 2.52 ( 1H, t, J = 4.4 Hz), 2.99 (1H, dd, J = 4.4, 7.0 Hz), 3.2 (1H, d, J = i4.7 Hz) j 3.29.3.42 (3H, m), 3.54 (1H , m), 4.13 (1H, d, J = 15.8 Hz), 4.30 (1H, dd, J = 6.2, 9.2 Hz). HRMS estimates cuHi7N207S, 333.0756. Found 333.0740. Example 5 Amino-4′-fluorenylthio-butadiene y2.2_di ^ ~ ^ ~~ ^ 1 ^ bicyclic "3.1.01Hexyl-4,6-dirollate_ 豳

Prepared according to step 3, using (11 ^, 43,58,63) -4- (2,8-third butoxy

85435 134- 200400815 Talking about the basic group -41-methylthio-butaminyl) _2,2_diketone_2λ6_congestion-bicyclo [3.1_0] hexane-4,6-dicarboxylic acid dimethyl Esters (077 g, 16 mmol, Preparation 12) yielded 0.41 g (54.9%) of the title compound. | &gt;] Lan = + 4 (c = 1.〇〇, Me〇H). ! H NMR (300 MHz, CD3OD) 52.1-2.2 (2H, m), 2.13 (3H, s), 2.47 (1H, t, J-4.4 Hz), 2.58-2.63 (2H, m), 3.02 (1H, dd, J = 4.0, 7.0 Hz), 3.12 (1H, d, J = 14.7 Hz), 3.52 (1H, dd, J = 3.3, 6.6 Hz), 3.98 (1H, t, J = 6.2 Hz), 4.14 (1H, d, J = 14.7 Hz). HRMS estimates C12H19N207S2, 367.0634. Found 367.0634. Example 6 -Amino group π-ΠΗ-AZADOL-3-a Λ-two life, its fluorene group I-2,2-diser-bicyclo "3.1.01 hexane-4.6-dimerium hydrochloride

Prepared according to general step C, using (1 foot, 43,58,63) -4- [2'8-third butoxycarbonylamino] third butoxycarbonyl-1Η-fluorin-3-yl) -propane Fluorenylamino] -2,2-difluorene-2λ6-thia-bicyclo [3 丄 0] hexane-4,6-dicarboxylic acid dimethyl ester (0.54 g '0.83 mmol, Preparation 13) can 0.28 g (73.6%) of the title compound was produced. [〇 + 7.8 (c = 1.02, CH3OH). 'Η NMR (300 MHz, CD3OD) δ 2.47-2.53 (1H, m), 3.05-3.18 85435 200400815 (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 = 7.7 Hz). Analysis estimates C18H19N307S * HC1: C, 47.22; H, 4.40; N, 9.18. Found: C, 46.51; H, 3.96; N, 8.54. MS (ES) m / z 420.1 [M-1], Example 7

Li ^ l ^! S, 6S) -4- 「2S-Amine-W4- 葙 葚-苽 some） -Bing Yimai 1 1 ^ _ ^ 1 is the same as -2 people 6-Poor-Second Ring" 3.1. 01 production, alkane-4.6-diphosphonate __

Prepare 'Application (ir, 4S, 5S, 6S) -4- [2, S-Third-butoxycarbonylamino-3'-(4-Third-butoxycarbonyloxy-phenyl) · according to General Step C · Propylamino] -2,2-dione-2λ6-pyrimidine-bicyclo [3.1.0] hexane_4,6-dicarboxylic acid dimethyl ester (0.33 g, 0.53 mmol, Preparation 14) can be generated 0.13 g (56.4%) of the title compound. [af〇 = -6 (c = 1.〇〇, H2〇) ° NMR (300 MHz, CD3OD) δ 2.47 (1H, t, J = 4.0 Hz), 2.87 (1H, dd, J = 9.2, 14.7 Hz ), 3.05-3.12 (2H, m), 3.23 (1H, dd, J = 5.1, 14.7 Hz), 3.55 (1H, dd, J = 4.0, 7.0 Hz), 4.00 (1H, dd, J = 4.8, 9.2 Hz), 4.13 (1H, d, J = 14.7 Hz), 6.80 (2H, d, J = 8.4 Hz), 7.13 (2H, d, 8.8 Hz). HRMS estimates C16H19N208S, 399.0862. Found 399,0844. 408 85435-136- 200400815 Examples &amp; (1R, 4S.5S λ. ~~ Amino-3 / -benzyl-propanylamine amine m -2¾6 + ^-~ U3.1.〇1 hexane-4.6 -Dicarboxylic acid hydrochloride

Prepared according to I and step c, using (111,48,58,68) _4_ (2,8_Third-butoxynylamino-3-benzyl-propionylamino) -2,2-dione -216-Pyrimidine-bicyclo [3.1.0] Hexahedral 4,6_di-chicoic acid dihydrazone (Preparation 5). Yield 0.55 g (85%, 1.31 milliwatts) as a white solid.

Wt + 4.17 (c = 0.48, MeOH). H NMR (300 MHz, CD3OD) δ 2.45 (1H, t, J = 4.0 Hz), 2.98 (1H, dd, J = 9.2, 14.7 Hz), 3.〇6 (1H, dd, J = 4.4, 7.0 Hz ), 3.10 (1H, d, J = 14.6 Hz), 3.33 (1H, dd, J = 4.8, 13.6 Hz), 3.52 (1H, dd, J = 3.3, 7.3 Hz), 4.08 (1H, dd, J = 5.1, 8.8 Hz), 4.10 (1H, d, J = 15.4 Hz), 7.31-7.42 (5H, m) 〇13C NMR (300 MHz, D2〇w / 1,4-dihumane): δ 172.55, 171.94 , 170.01, 134.31, 130.39, 130.27, 130.01, 129.00, 61.38, 54.76, 54.37, 43.07, 37.36, 31.57, 23.18. Analysis estimates C16H18N207S &lt; .5 HC1: C, 43.97; Η, 4.50; N, 6.41. Found: C, 43.59; H, 4.17; N, 6.46. MS (ES) m / z found 383.1 [M + H] +. 85435 • 137- 200400815 HRMS estimate c16Hl9N207S [M + H] +: 383.091 3. Found: 383.0923. Example 9 Amine_3’s_methyl-pentanamine_) 2,2_dione Bicyclo Γ3.1.01 Hexane-4,6-di # _ Hydrochloride

V

Prepared according to general step c, using (111,43,53,6 phantom_4_ (2,8_third butoxyalkylamino-3'-methyl-pentamylamino) _2,2_dione _2λ6_3Ser-bicyclo [3 丄 〇] Jijian * _4,6 · Dicarboxylate (Preparation 6). Yield 0.43 g (80%, 1.12 mmol) white solid. [A] g = +4.08 (c = 0.49, MeOH).! H NMR (300 MHz, CD3OD) δ 0.98 (3H, t, J = 7.3 Hz), 1.07 (3H, d, J = 7.0 Hz), 1.15-1.24 (1H, m), 1.53-1.62 (1H, m), 1.95-2.04 (1H, m), 2.44 (1H, t, J = 4.0 Hz), 3.06 (1H, dd, J = 4.4, 7.0 Hz), 3.14 (1H , d, J = 15.0 Hz), 3.52 (1H, dd, J = 3.7, 7.0 Hz), 3.73 (1H, d, J = 5.1 Hz), 4 · 10 (1H, d, J = 14.7 Hz). Analyzed and estimated C13H2ON207S * HC1 * 0.5 H2O: c, 39.64; H, 5.63; N, 7.11. Found: C, 39.3 8; H, 5.39; N, 7, 04. MS (ES) m / z measured Value 349.1 [M + H] +. HRMS estimate C13H21N207S [M + H] +: 349.1069. Found: 349.1086. 85435 -138- 200400815 Example 10 Amine-3 ^ methyl-butylum-some amino)- 2,2-diketone ^^^-dioxan.1.01 hexane-4.6-dihydrochloride

Prepared in the same manner as in step C, using (ir, 4S, 5S, 6S) -4- (2, S-tertiary butoxyalkylaminomethyl-butylamidinoamino) -2,2-dione-2λ6- 4-Bicyclo [3.1.0] hexane-4,6-dicarboxylic acid dimethyl ester (Preparation 7). Yield: 0.18 g (88%, 0.49 mmol) as a white solid as HC1 salt. [0; H = + 7.84 (c = 0.51, MeOH). 'η NMR (300 MHz, CD3OD) δ 1.04 (3H, d, J = 6.6 Hz), 1.09 (3H, d, J-7.〇Hz), 2.22-2.29 (1H, m), 2.44 (1H, t , J = 4.0 Hz), 3.05 (1H, dd, J = 4.4, 7.0 Hz), 3.10 (1H, d, J = 14.7 Hz), 3.52 (1H, dd, J = 4.0, 7.3 Hz), 3.67 (1H , d, J = 5.5 Hz), 4.10 (1H, d, J = 14.7 Hz). Analytical estimates (: 1211181 '«12〇73 # 〇1 * 0.4 112〇: (:, 38.12; 11, 5.20; 1 ^, 7.41. Found: C, 37.78; Η, 4.90; N, 7.15. MS (ES) m / z Found 335.1 [M + Η] +. Example 11 (1R, 48.58,68) -4- (23-Amine-41-Methyl-Amine-Amine) -2,2- Erli 85435 139- 200400815 illi-fluorene-bicyclo "3.1.01 hexane-4.6-dimiao_phosphonate

Prepared according to general procedure C, using (111,48,58,68) -4- (2,3-third butoxycarbonylamino-4, · methyl-pentamylamino) -2,2-di Keto-2λ6-pyrimidine-bicyclo [3.1.〇] hexane-4,6-dicarboxylic acid dimethyl ester (Preparation 8). This gave 0.50 g (76%, 1.30 mmol) of a white solid as the HC1 salt. [〇 ^ 3 = -4.0 (c = 0.50, MeOH). NMR (300 MHz, CD3OD) δ 1.00 (3H, d, J = 5.5 Hz), 1.02 (3H, d, J-5.9 Hz), 1.62-1.79 (3H, m), 2.42 (1H, t, J = 4.0 Hz), 3.04 (1H, dd, J = 4.4, 7.3 Hz), 3.13 (1H, d, J = 15.0 Hz), 3.52 (1H, dd, J = 3.3, 7.0 Hz), 3.84-3.89 (1H, m ), 4.10 (1H, d, J = 15.0 Hz). Analysis estimates C13H2ON207S * HCUO_3 H20: c, 40,01; H, 5.58; N, 7.18. Found: C, 39.66; H, 5.57; N, 6.99. MS (ES) m / z found 349.1 [M + H] +. Example 12 Di-SiJi-based amine and even Λ-2.2-dione-2λ6-pyrimidine-bicyclo [3.1.01P, fe-4,6-dicarboxylic acid _ gj 85435 -140-

200400815 According to the same steps, prepared in step c, using (1 foot, 48,53,68) -4- (2'3,6, -bis-third butoxycarbonylamino · hexamidineamino) _2,2 _Dione_2λ6_pyrimidine_bicyclo [3. [Chuanane 4,6-dicarboxylic acid dimethyl ester (Preparation 9). It produces 0.56 g (86%, 1.28 mmol) of a white solid as a double hydrochloride salt. [a] g = -4.0 (c = 0.50, MeOH). 'H NMR (300 MHz, CD3OD) δ 1.48-1.55 (2H, m), 1.67-1.74 (2H, m), 1.89-1.97 (2H, m), 2.47 (1H, t, J = 4.0 Hz), 2.97 (2H, aPP · t, J = 4.0), 3.08 (1H, dd, 3 = 4.4, 7.0 Hz), 3.20 (1H, d, J ^ IS.O Hz), 3.53 (1H, dd, J = 3.7, 7.0 Hz), 3.93 (1H, app. T, J = 6.2 Hz), 4.08 (1H, d, J = 14.7 Hz). Analysis estimates C13H21N3O7S * 2HCN0.2H20: C, 35.49; H, 5.36; N, 9.55. Found: C, 35.30; H, 5.48; N, 9.42. MS (ES) m / z found 364.1 [M + H] +. Example 13 1111,48,58,68)-4-] ^ | 8-Amino-4 | -Aminomethyl-butyroylamino) _2,2-diketone-2λ6-ρΐ · -bicyclo `` 3.1.0 ~ | Hexane-4.6-Diquinic Acid Pound 85435 -141-200400815 V / 0

(lR, 4S, 5S, 6S) -4- [2'S-Third-butoxycarbonylamino-4,-(trityl-aminomethyl) -butylimidoamino] -2,2-difluorene- 2 persons 6-1 »Ser-bicyclo [3.1.0] hexane-4,6-dicarboxylic acid dimethyl ester (0.48 g, 0.65 mmol, Preparation 10) in 1: 1 2.5 N LiOH and THF (6 ml total volume) The mixture was stirred at room temperature for 4 hours. The reaction mixture was adjusted to pH = 2 with 1 N HC1, and the product was extracted with ethyl acetate. All organic layers were mixed, washed with brine ', dried over MgSCU and concentrated to yield 0.46 g (1 foot, 48,58,63) -4- [2 | 3-third butoxycarbonylamino-4,-(triben Methyl-aminomethylamino) -butylaminoamino] -2,2-dione-2λ6-tetra-bicyclo [3 · ΐ〇] hexane-4,6-dicarboxylic acid 'was a white foam. The diacid was dissolved in DCM and stirred at room temperature 'followed by the addition of anisole (0.28 g' 2.6 mmol) and tfa (3 g '32.5 mg). The resulting reaction mixture was stirred at room temperature for 2 hours and then concentrated under vacuum. The resulting yellow oil was ground in EGO until it formed a free-flowing white precipitate. TFA was collected in a Nz basket by suction filtration. The product was dissolved in 1 ml of 1 NHC1 and subjected to freeze-drying to produce the desired product as HC1 salt. This gave 0.16 g (62%, 0.40 mmol) of a white solid. [a] 2fl3 = + 8_0 (c = 1.0, H2〇). a] H NMR (300 MHz, CD3〇D) δ 2.00-2.08 (2H 〇,, in jz j 7 _ 2 41 -142- 4 | 4 85435 200400815 (3H, m) 5 2.93 (1H, dd, J = 4.3, 7.3 Hz), 3.04 (1H, d, J = 14.7 Hz), 3.45 (1H, dd, J = 3.7, 7.0 Hz), 3.86 (1H, app. T, J = 5.9 Hz), 4.05 (1H, d, J = 14.7 Hz). Analytical estimation &lt;: 12111 屮 3088 ((1 * 2.0 1120,. €, 33.07; Fan 5.09 ;: ^, 9.64. Found: C, 33.37; H, 4.69; N, 9.39. MS (ES) m / z found 363.9 [M + H] +. HRMS (ES) estimated C12H18N308S [M + H] +: 364.0815. Found: 364.0825 〇 Example 14 (1R, 2S, 4R, 5R, 6R) -2- (2'S · Amine-Propanyl) -Amine 4-Gas Bicyclo [_3-Di 1.01 Hexane-2.6-Di # Acid Test

(1 foot, 28,411,511,611) -4- (2 | 8-2 '-(Third-butoxyamino-propylamino) propyl) Amino-4-fluoro-bicyclo [3 · 1 · 〇] hexane_2,6-dicarboxylic acid (53 g corrected for acetone '142 mmol) in a slurry of 447 ml of acetone, stirred in 5 (rc acetone for 3 5 minutes. The hazy solution was filtered to Clarify 'and rinse with 100 ml of acetone. 22.1 ml (265 mmol) of concentrated hydrochloric acid is added dropwise to the clear and white-doped filtrate for 5 days. The mixture is warmed to 45-50 ° C, And incubate for 90 minutes. The mixture is implanted with the instrument as required (23,411,511,611) _2_ (2,8_aminopropylamido) amino-4-fluorobicyclo [3.1.0] hexane_ 2,6_dicarboxylic acid hydrochloride, then stopped heating, and then gradually cooled to room temperature. After 2 hours the temperature reached 25t, and then added acetone (942 41S 85435 -143-200400815 ml) to the slurry for 90 minutes. The slurry was stirred for another 16 hours, then filtered, washed with acetone (2x200 ml), dried under vacuum at 45 ° C for 9 hours, and again at room temperature for 64 hours to give 40.2 g (91%) of the title compound as a white solid. Recrystallization 1.06 g dissolved in 0.5 ml water and 2.1 2 ml of acetone, heated at 50 ° C, then diluted with 5.3 ml of acetone, and then seeded as needed. In this slightly turbid mixture, add 4.2 ml of acetone as needed, and seeded again, turn off Heat source, let it gradually cool to room temperature for 1 hour. Dilute the resulting slurry with another 9.5 ml of acetone for 30 minutes and stir for 15 hours. Once filtered, wash with acetone (2 x 5 ml) at 45 ° C Dry for 10 hours under vacuum and 60 hours at room temperature to obtain 0.905 g (85% recovery) of the title compound as a white solid. Mp (DSC) 183 ° C. [A] 25D +33. (C 1.06, CH3OH). 500 MHz,! H NMR (CD3OD) δ 5.58-5.42 (m, 1H), 3.92 (q, 1H, J = 7.0 Hz), 2.96 (dd, 1H, J = 14, 8.0 Hz), 2.41- 2.39 (m, 1H), 2.35-2.30 (m, 1H), 2.10 (t, 1H, J = 3.0 Hz), 1.52 (d, 3H, J = 7.5 Hz), 1.51-1.42 (m, 1H). 13C NMR (125 MHz, CD3OD) δ 173.74, 173.62, 170.00, 93.48 and 92.04 (CF split), 63.95 and 63.92 (CF split), 48.80, 36.89 and 36.70 (CF ^ f.), 32.97 ^ 32.91 (CF ^ f.), 30.05 ^ 129.87 (CF split), 19.37, 16.28. FTIR (DRIFT) 3430 (w), 3016 (s), 1721 (s), 1662 (s), 1496 (s), 1190 (m), 1024 (m), 637 (w) cm-1. 85435 • 144 · 200400815 Meeting Example 15 UR, 2S.4R.5R, 6RVm, S-Amine-Propanamine) · Amino-4-fluorobicyclo 『3.1.01P, Alkane-2.6-Di # Acid Methanesulfonate Acid salt

(111,28,411,511,6 feet) -2- [2'8- (third butoxycarbonylamino) propanyl] -amino_4-fluorobicyclo [3.1.0] hexane-2, A slurry of 6-dicarboxylic acid (1.87 g corrected, 4.98 mmoles' Preparation 18) in 16.8 ml of acetone was brought to 50. (: Stir for 15 minutes. Filter the hazy solution to clarify the solution, then rinse with acetone (3 x 1.25 ml). The clear filtrate is diluted with 0.935 ml of water and placed in a hot water bath of 5 (TC). And dropwise treatment with 0.647 ml (9.97 mmol) of methanesulfonic acid (gas evolution can be observed). A white slurry will be produced after 25 minutes. After stirring for 2 hours, add another 5-10 minutes 35 5 ml of acetone. Stop heating and allow the slurry to slowly cool to room temperature for 2 hours, then filter, wash with acetone (2x8 ml), and dry at 45 ° C for 14 hours to produce 177 g ( 95%) of the title compound as a light pink solid. A sample of this material was recrystallized as follows: 1.65 g dissolved in 1.16 ml of water and 4.95 ml of acetone, heated at 50 t, and seeded as needed. Stop Add acetone (26.4 hours at the same time for 40 minutes. Once filtered, acetone (2x6 at room temperature for 60 hours, obtain 1.59), then dilute with another 1.65 ml of acetone, and let the mixture gradually cool to room temperature depending on the heat. At the same time, the resulting slurry was stirred for another 3 hours. Wash, vacuum dry at 45 ° C and dry in a chamber 85435 -145-200400815 g (96% recovery) of the title compound as a white solid. Mp (DSC) 206〇C 0 [a] 25D +30 (c 1.05 , CH3OH). * H NMR (500 MHz, CD3OD) 55.58-5.42 (m, 1H), 3.92 (q, lH? J = 7.0 Hz), 2.96 (dd, 1H, J = 14, 8.0 Hz), 2.70 ( s, 3H), 2.41-2.39 (m, 1H), 2.35-2.30 (m, 1H), 2.10 (t, 1H, J = 3.0 Hz), 1.52 (d, 3H, J = 7.5 Hz), 1.51-1.42 (m, 1H). 13C NMR (125 MHz, CD3OD) δ 173.73, 173.61, 170.02, 93.50 and 92.65 (CF split), 63.91, 48.79, 38.30, 36.89 and 36.70 (CF split), 32.97 and 32.91 (〇? split ), 30.02 and 29.84 ((3- 卩 division), 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. The analysis estimates C12H19FN208S: C, 38.92; H, 5.17; N, 7.56. Measured value; C, 38.96; H, 4.97; N, 7.51 ° Example 16 〇 ^, 28.4 feet. 5.! 1.6 ^ 〇-2- (2 | 8-amino-propyl hydrazine) amine-4--4- | Dioxin "3.1.0_1 PV grab.-2.6-dicarboxylic acid acetamate

(1 ', 28,4', 5 ', 611) -2- [2 $-(Third-butoxycarbonylamino) propanyl] -amine.146- 41 «85435 200400815 4-fluorobicyclo [3.1.0] A slurry of hexane-2,6-dicarboxylic acid (0.2 g, 0.534 mmol, Preparation 18) in 1.8 ml of acetone, and allowed to stir at 50 t for 5 minutes. Pass the cloudy solution to clarify the solution, and then rinse with acetone (1 x 04 real liters). The clear filtrate was diluted with 0.1 ml of water, placed in a 5 (TC heating bath, and then treated with 0.124 ml (1.07 mmol) of acetic acid in a dropwise manner (gas evolution was observed). It can be generated in 90 minutes White slurry. After stirring for a total of 2 hours, add another 1.8 ml of propane I for 5 minutes. Stop heating, and gradually cool the slurry to room temperature for 1 hour 'and stir for another 2 hours. Filter and wash with acetone (2X1 ml) After drying under vacuum at 45 ° C for 4 hours, and then at room temperature for 60 hours, 0.173 g (84%) of the title compound was formed as a white solid. Mp (DSC) 210 ° C (decomposition). ! H NMR (500 MHz, CD3〇D) δ 5.58-5.42 (m, 1H), 3.92 (q, 1H, J = 7.0 Hz), 2.96 (dd, 1H, J = 14? 8.〇Hz), 2.80 (q, 2H, 7.3 Hz), 2.42-2.37 (m, 1H), 2.35-2.30 (m, 1H), 2.09 (t, 1H, J = 3.0 Hz), 1.52 (d, 3H, 1 = 7.5 Hz) , 1.51-1.40 (m, 1H), 1.30 (t, 3H, J = 7.5

Hz). Example 17 "Umin" ^ 4g ^ iR, 6R) -2- (2’§J1 amine jade-propanone) A bicylic ice-gas ring [· 3.101 fe * 2 ′ g reading

F

85435 -147- 200400815 (1 foot, 23,4 feet, 511.6 feet) -2- [2'8- (third butoxycarbonylamino) propanyl] -amino-4-fluorobicyclo [ 3.1.0] A slurry of hexane-2,6-dicarboxylic acid (0.402 g, .07 mmol, Preparation 18) in 3.6 ml of acetone and brought to 50. (: Stir down for 10 minutes. Treat the dim solution with a small amount of celite to clear the solution, then rinse with acetone (2x0.4 ml). Place the clear filtrate in a 5 (Tc hot water bath, Treatment of 226 mg (90% '1.29 mmol) of benzenesulfonic acid in 0.113 ml of water' Follow with 0.4 ml of acetone (gas evolution can be observed). 4 hours after stirring under gentle reflux ' The heating was stopped and the reaction was treated with 8 ml of acetone for 10 minutes, and seed crystals were added as needed. A slurry formed in 1 hour, which was then diluted with 3.2 ml of acetone 'followed by stirring at room temperature for another 15 5 hours. Filtered 'Washing in acetone (2x10 ml) and drying it at 45 ° C for 24 hours yielded 313 mg (62% corrected for 10 wt% acetone) of the title compound as a white solid. Mp (DSC) 132 ... NMR (500 MHz, CD3OD) δ 7.86-7.80 (m, 2H), 7.46-7.37 (m, 3H), 5.58-5.42 (m, 1H), 3.92 (q, 1H, J = 7.0 Hz), 2.96 (dd, 1H J = 14, 8.0 Hz), 2.42-2.37 (m, 1H), 2.35-2.30 (m, 2H), 2.09 (t 1H, J = 3.0 Hz), 1.52 (d, 3H, J = 7.5 Hz), 1.5U1.40 (m, iH). Example 18 (1 R, 2S, 4R, 5R, 6R) _- 2- (21S-amino-propanyl) 4-fluoro-ring "3 1 1 hexane-2,6-dicarboxylic acid benzenesulfonate

NH2. PTolS〇3H 85435 • 148- 200400815 (lR, 2S, 4R, 5R, 6R) -2- [2'S- (third butoxycarbonylamino) propanyl] -amino-4-fluorobicyclo [ 3.1.0] A slurry of hexane-2,6-dicarboxylic acid (1.04 g corrected, 2.78 mmoles' of Preparation 18) in 9,36 ml of acetone, which was stirred off under 15 minute. The hazy solution was treated with a small amount of celite, filtered to clarify the solution, and then rinsed with acetone (1 x 2.08 ml, and then 1.04 ml). The clear filtrate was placed in a heating bath at 50 ° C, treated with a solution of 634 mg (3.33 mmol) of para_toluenesulfonic acid monohydrate in 0.3 17 ml of water, and then rinsed with 0.317 ml of acetone (may Outgassing was observed). After stirring for 4 hours under gentle reflux, the reaction was removed from the heating bath and treated with 10.4 ml of acetone for 10 minutes. The clear and colorless solution was seeded as needed, and the formation of a precipitate was observed after 30 minutes. At this time, another 10.4 ml of acetone was introduced in 20 minutes. The slurry was allowed to stir for another 4 hours, then filtered, washed with acetone (2 ml), and dried under vacuum at 45 ° C for 14 hours to yield 995 mg (78% corrected for 3 wt% acetone) of the title compound as white solid. mp (DSC) 155 ° C. lU NMR (500 MHz, CD3OD) δ 7.70 (d, 2H, J = 7.5 Hz), 7.34 (d, 2H, J = 8.5 Hz), 5.58-5.42 (m, 1H), 3.92 (q, 1H, J = 7.0 Hz), 2.96 (dd, 1H, J = 14, 8.0 Hz), 2.42-2.30 (m, 2H), 2.24 (s, 3H), 2.09 (t, 1H, J = 3.0 Hz), 1.52 (d, 3H, J = 7.5 Hz), 1.51-1.40 (m, 1H). Example 19 (1R, 2, S, 4Amine-Propanyl) Amine-4-Affinity, Bicyclic "3.1.01 Hexane-2,6-di # acid

85435 -149-

F

200400815 p- (111,28,411 wind 611) _2_ (2,8_amino_propanyl) _amino_4-gasbicyclo [3 · 1.〇] hexane; k_2,6 · dicarboxylic methanesulfonic acid Salt (ο. 5 g, 13S millimoles, Example 15), 1 liter of water, solution, add 5 ml U ethanol at 5 () t, and add G. 27 ml (1.35 mmol) after a few minutes Mol) 5 sulphuric acid solution. Stop heating and dilute the clear, colorless solution with 2.5 ml of ethanol, add seed crystals as needed, and then dilute further with 7.5 ml of ethanol for ㈣ minutes. The resulting slurry was stirred and cooled at room temperature for 1 hour after counting to f for 2 hours at a high temperature. The solid was collected, washed with ethanol (1 x 10 ml), and dried at 45 ° C under vacuum for 18.5 hours to yield 0.301 g (78% yield, calculated as 1.6 wt% sodium methanesulfonate and 3 wt% ethanol corrected) of the title compound ,? , 疋 white solid. lH NMR (500 MHz, D20) δ 5.45-5, • J〇 (m, 1Η), 3.88 (n in J = 7.0 Hz), 2.58 (dd, 1H, J = 14, 8.〇 u,

Hz), 2.33-2.30 (m) 2.27-2.26 (m, 1H), 1.92 (t, 1H, J = 3)

Wz), 1.36 (d, 3H, J = 7 l

Hz), 1.41-1.32 (m, 1H). 13C NMR (125 MHz, D20) δ 177.46 ~ '176.92, 17G.42, 94.56 and 93.19 (OF split), 65.36, 49.〇1, 36.75 and 36 57 (ο split 33.61 and 33.55 (CF split 30.54 and 30.36 (C_F cleavage), 20A 16.67. Example 20 85435 150- 200400815 U_R, 2S, 4125 R, 6-based_propanyl) Amine_4_fluorobicyclic "v 彳

~ ~ I 二 # Acid drowsiness

P- (111,28,411 wind 6 feet) -2- (2,8-amino_propanyl) _amino_4_fluorobicyclo [3.1.0] hexane-2,6-dicarboxylic acid methanesulfonate Acid salt (70 mg, 019 mmol, Example 17) in 420 µl of methanol in 6 (rc) solution, and sodium acetate (46 5 mg '0.57 ¾ mole) was added to 470 µl of a warm solution of methanol Rinse with 230 microliters of methanol. Stop heating. Dilute the stirred solution with 28 microliters of methanol and add seed crystals as needed to aid crystallization. The resulting slurry is slowly cooled to ambient temperature After 1 hour, stir at ambient temperature for 2 hours. The product was collected by filtration, washed with methanol (2x280 microliters), and dried at 45. (3 under vacuum for 15 hours to 52.5¾ g (85% yield, for 2.3 wt% sodium formate and 0.2 wt% methanol correction) The title compound was a white solid. H NMR (500 MHz, D20) δ 5.44-5.29 (m, 1Η), 3.89 (q, 1H, J = 7 .〇Hz), 2.65 (s, 3H), 2.56 (dd, 1H, J = 14, 8.0 Hz), 2.16-2.13 (m, 1H), 2.10-2.09 (m, 1H), 1.74 (t, 1H, J-3.1 Hz), 1.38 (d, 3H, J = 7.1 Hz), 1.36-1.28 (m, 1H). 13C NMR (125 MHz, D20) δ 180.00, 178.72, 170.13 , 95.40 and 93.99 (CF split), 65.97, 49.06, 37.25 and 37.07 (CF split), 33.01 and 32.94 ((:-? Split), 29.64 and 29.46 (〇? Split), 22.48, 85435 -151-200400815 16.68. Example 21 ^ • ^ lg, 5R, 6R) -2-(^ S-amine, propionate, amine), amine, 4-hydroxyline, -difluorene, 1.01 hexane-2,6-di # hydrochloride

H3C NH2. HCI 〇8,28,48,5 feet, 611) -4-Ethyloxy-2- [2 | 3- (third butoxy) carbonylaminopropylamido] -aminodiamine Cyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester (0.600 g '128 mmol, Preparation 21) in THF (30 mL) and water (30 mL) was added to the solution. Lithium hydroxide monohydrate (0535 g, 128 mmol). After stirring at room temperature for 3 hours, the reaction was diluted with water and washed with ethyl acetate (2X25 * L). The aqueous layer was acidified to pH 1 with 1 N HC1 and extracted with ethyl acetate (3x30 mL). The combined organic extracts were concentrated, redissolved in 1 n hci / ethyl acetate (60 ml), and stirred at room temperature under nitrogen for 16 hours. The solvent was removed under vacuum to yield 400 mg (99%) of the product as a white solid. LCMS: m / z 273 [M + H] + @Rt 0.20 minutes. (YL NMR (CD3OD) *: 4.13 (1H, d, 5.9 Hz), 3.80 (1H, q, 6.7 Hz), 2.41-2.31 (2H, m), 1.93 (1H, dd, 6.0 Hz, 2.7 Hz), 1.62 (1H, dd, 5.9 Hz, 15.5 Hz), 1.44 (1H, t, 3.0 Hz), 1.38 (3H, d, 6.8 Hz); * NB exchangeable protons cannot be observed with 1 ^? ^ 11 = 6 . Example 22 85435 -152- 200400815 (lg5 amine and even acetamido) -4-pyridyl_bicyclo "3 〇 ^ alkane-2,6-dihydrochloride

Prepared according to general step C, using (13,23,48,5 feet, 611) -2- (2'-third butoxycarbonylamino-ethylamine) _4-meryl-bicyclo [3.10] hexane Burning diethyl 2,6-dicarboxylic acid (300 mg, 0-84 millimoles, preparation 23), with the exception that the removal of the third butoxycarbonyl protecting group was treated with 4MHC1 /: Yield: 156 mg (63%). [036 (c = 0.5, MeOH). 5H NMR (400 MHz, CD3OD) δ 1.56 (1H, t, J = 2.9 Hz), 1.74 (1H, dd, J = 5.8, 15.4 Hz), 2.12 (1H, m), 2.48 (1H, d, J = 15.4 Hz), 2.61 (1H, m), 3,62 (2H, s), 4.32 (1H, d, J = 5.49 Hz). Analysis estimates C1 () H14N206M.3 HCUH20: C, 37.11; H, 5.39; N, 8.66. Found: c, 37 · 36; H, 4.99; N, 8.30. MS (ES) m / z 258.8 [M + H] +. Example 23 (1§ ^, 48,511, 610-2- (2'8-amino-3 | -methyl-butane_its dizziness) -4_ 衮 -bicyclic Π.ί · 〇1 Jujube 35 85435 • 153- 200400815

ch3 = general step c, using (1s, 2S, 4S, 5R, 6r) _2_ (2is1 ^ = fluorenylamino ~ 3'-methyl-butanylamino) -4-hydroxy-bicyclo [3.1. 0] Hexane • 2,6-dicarboxylic acid diethyl ester (48 mg, 119 mmol, prepared as), except for the removal of the -butoxycarbonyl protecting group as 4 M HC1 / 二 # Calcined, yield 307 mg (76%). [&lt; = + 8.33 (c = 0.48, Me0H). 'Η NMR (400 MHz, CD3OD) δ 1.07 (3H, d, J = 6.8 Hz), 1-Π (3H, d, J = 7.3 Hz), 1.55 (1H, t, J = 2.9 Hz), 1.76 ( 1H, dd, J = 5.8, 15.6 Hz), 2.14 (1H, dd, J = 3.4, 5.8 Hz), 2.24 (1H, m), 2.50 (1H, d, J = 15.6 Hz), 2.64 (1H, dd , J = 2.9, 5.8 Hz), 3.66 (1H, d, J = 5.4 Hz), 4.32 (1H, d, J = 5.8 Hz). Analysis estimates C13H2 () N206 * HCN1.1 H20: C, 43.79; H, 6.56; N, 7.86. Found: C, 43.77; H, 6.20; N, 7.47. HUMS estimates C13H21N206, 301.1400. Found: 301.1400. f Example 24 (IS, 2S. Amine amino group)-4 trace production-a hindrance "3.1.01 hexane-2, ^ · ^ · ^^ 85435 -154-

200400815

According to the procedure of Step C, (1S, 2S, 4S, 5R, 6R) -2- (2, S1: T oxoamido-4'-fluorenyl-pentamido) -4-hydroxy- Dicyclo [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester (46 mg, m mmol, preparation 25), except for the removal of the second butoxycarbonyl protecting group was 4 M HC1 / Dizane treatment. The yield was 371 mg (95%).

Wg = + 4 (c = 0.5, MeOH). 'Η NMR (400 MHz, CD3OD) δ 1.01 (3H, d, J = 5.8 Hz), I.03 (3H, d, J = 6.3 Hz), I.54 (1H, t, J = 2.9 Hz), 1.63-1.82 (4H, m), 2.14 (1H, dd, J = 2.9, 5.8 Hz), 2.49 (1H, d, J = 15.6 Hz), 2.62 (1H, dd, J = 2.9, 5.8 Hz), 3.83 -3.86 (1H, m), 4.32 (1H, d, J = 5.g Hz). Analysis estimates H20: C, 44.72; H, 6.92; N, 7.45. Found: C, 44.52; Η, 6.57; N, 7.13. HRMS estimates ChH23N206, 3 15.1556. Found: 3 15.1569. Paste Example 25 .f-Pentamidine) -4- 葙 I: Bicyclic "3.1.01 Hexane-2,6-dicarboxylate 酩 85435 -155- 200400815

OH

ch3 was prepared in the same manner as in step C, using (1S, 2S, 4S, 5R, 6R) -2- (2'S-fST oxenylamino-3's-methyl-pentamidine) -4-hydroxy-bicyclo [3.1.0] Hexane-2,6-diacetic acid diethyl ester (41 mg, 099 mmol, Preparation 26), except for the removal of the third butoxycarbonyl protecting group was 4 M HC1 / Dioxane treatment. The yield was 330 mg (95%). [0 + 8 (c = 0.5, MeOH). 'Η NMR (400 MHz, CD3OD) δ 0.99 (3H, t, J = 7.3 Hz), 1.08 (3H, t, J = 7.3 Hz), 1.17-1.27 (1H, m), 1.53 (1H, t, J = 2.9 Hz), 1.59-1.65 (1H, m), 1.76 (1H, dd, J = 5.8, 15.6 Hz), 1.96-2.00 (1H, m), 2.14 (1H, dd, J = 2.9, 5,8 Hz), 2.46 (1H, d, J = 15.6 Hz), 2.66 (1H, dd, J = 2.9, 5.8 Hz), 3.70 (1H, d, J = 5.4 Hz), 4.31 (1H, d, J = 5.8 Hz). Analysis estimates C14H22N206 * 1.1 HC1M.2 H20 ·· C, 44.71; H, 6.83; N, 7.45. Found: C, 44.3 8; H, 6.51; N, 7.08. HRMS estimates C14H23N206, 3 15.1 556. Found: 3 1 5.1 566. f Example 26 £ 1 ^ 8,48,511,6 feet) -2 di [2 | -〇amino-ethyl alcohol amine) -ethyl_some amine 1-4_ hydroxyl-bicyclo 丨 3.1.01 hexane-2 -6- Ermiao 428 85435 -156- 200400815

Treated with an excess of 4 M HC1 / dioxane (is, 2S, 4S, 5R, 6R) -2- [2,2 second butoxyamino-ethoxyamine] _ethylamine] _4_ Ethyl-difluorene [3.1.0] hexane-2,6-dicarboxylic acid diethyl ester (360 mg, 0.77 mmol, Preparation 27), and stirred for 40 minutes' and concentrated under vacuum. Ethyl acetate was added and concentrated. The solid was treated with THF (3 M) and 1 N LiOH (3.0 equiv.). After stirring for 1 hour, add IN HC1 until pH = 3. The reaction was concentrated under vacuum. Purified via DOWEX® 50WX8-100 ion exchange resin. Yield 8 mg (78%). [Cheng 3 = + 3.92 (c = 0.51, H20).

'Η NMR (300 MHz, D2O) δ 1.43 (1H, t, J = 2.9 Hz), 1.53 (m dd, J = 5.9, 5.4 Hz), 1.98 (1H, m), 2.23-2.31 (2H, m) , 3.70 (2H s), 3.82 (2H, app d, J = ll Hz), 4.16 (1H, d, J = 5.9 Hz). HRMS estimates Ci2H! 8N307, 3 16.1145. Measured value · 3 16.1123. Example 27 (18,28,43,5 feet, 6! 1) -2- "2 |-(28-Amine-Propanamide A) _Ethyl-Amine 4-1-4hydroxy-bicyclo" 3.1. 01 Hexane-2.6- Ermiao-157- 85435 200400815

r

According to Example 26 --- Propanamido) -Acetoamido] _ 4-hydroxy-bicyclo-ethyl (410 mg, 0.84 millimoles, Preparation-General procedure) (2S-Third-butoxycarbonylamino_ Propylamidine [3.1.0] Hexyl-2,6-dicarboxylic acid diethyl was used (1S, 2S, 4S, 5R, 6R) -2- [2,-28). Yield 200 mg (72%). [«Hyun = + 23.53 (c = 0.5.i, h2〇). H NMR (300 MHz, D20) δ 1.36 (3H, d, J = 7.0 Hz), 1.37 (1H, t, J = 3.3 Hz ), 1.49 (1H, dd, J = 5.9, 15.4 Hz), 1.91 (1H, dd, J = 2.9, 5.9 Hz), 2.15 (1H, dd, J = 2.9, 5.9 Hz), 2.27 (1H, d, J = 15.4 Hz), 3.79 (2H, s), 3.95 (1H, q, J = 7.0 Hz), 4.13 (1H, d, J = 5.5 Hz). HRMS estimates Cl3H19N307, 330.1301. Found: 330.1290. Example 28 nSJS / SjmiQJ-CS-Amino-3'-phenyl-propanamidine) -4-A-bicyclon.1.01 dane-2,6-dicarboxylic acid hydrochloride

HCI slightly 85435 -158- 200400815

Prepared according to general step C. (1s, 2s, 4S, 5R, 6r) _2- (2, s ^ sT oxycarbonylamino-3'-phenyl · propionamido) _4_hydroxy-bicyclo [3 ·]] Hexane-2,6-monocarboxylic acid monoethyl g (400 mg, 0.89 mmol, Preparation 29). Yield: 290 g (85%). Shout +3.64 (c = 0.55, MeOH). H NMR (400 MHz, CD3〇D) δ 1.45 (1H, t, J = 3.3 Ηζ), 1.60 (1H, dd, J = 5.5, 15.4 Hz), 2.03 (1H, dd, J = 2.6, 5.9 Hz), 2.37 (1H, d, J = 15.4 Hz), 2.55 (1H, dd, J = 2.9, 5.9 Hz), 2.90 (1H, dd, J = 8.4, 14.3 Hz), 3.22 (1H, dd, J -5.1, 14.7 Hz), 3.97 (1H, dd, J-5.5, 8.8 Hz), 4.21 (1H, d, J = 5.5 Hz), 7.19-7.31 (5H, m). HRMS estimates C17H2iN206, 349.1400. Found: 349.1388. Analyze Ci7H21N206 * HCWH20: C, 50.69; 5., 5.76; N, 6.95; Cl, 8-80. Found: c, 50.66; Η, 5.65; N, 6.85; Cl, 8.20. MS found 349.0 [M + H] +. Example 29 Amino-41-aminoformamidine-acid, amine and even _4jAlthyl-dibad "3 · 1.01 hexane, fe-2,6 · bisammonate

OH

The title compound was prepared from (18,28,48,5 feet, 6 Chinese) -2- [2-third-butoxy 4 3 ί 85435 -159- 200400815 tertamino-4-(diphenylmethyl- Carboxymethyl) butyrylamidoamino] -hydroxyl-dinon [3 1_0] Hexyl U, 6-dichloroacetate (118 mmol, Preparation 30) is dissolved in THF (H) ml, And treated with 25 N⑽ 叩 2 亳 liter). Allow the solution to stir for $ hours, then adjust the pH to 2 with 2.5 Να. The product was extracted four times with ethyl acetate, dried over sodium sulfate, and concentrated under vacuum to produce a white powder. The diacid product (0.74 mmol) was dissolved in dichloroethane (3.7 ml) and treated with anisole (0.3 ml) and trifluoroacetic acid (3.72 ml). The reaction mixture was stirred at 23 C for 2.5 hours and then concentrated under vacuum to give a brown oil. The brown oil was dissolved in water and extracted 5 times with dichloromethane. The aqueous layer was concentrated under vacuum and treated with 1 N HC1 (0.74 ml) and freeze-dried. The solid was treated with another ^ ν HC1 (2¾ liters) and cold-dried to yield 446 mg (quantitative) of the title compound. [T 3 = + 8.16 (c = 0_49, MeOH). lH NMR (400 MHz, CD3〇D) δΐ.60 (1H, t, J = 3.3 Hz), 1.79 (1H, dd, J = 5.5, 15.4 Hz), 2.10-2.17 (3H, m), 2.45-2.55 (3H, m), 2.60 (1H, dd, J = 2.9, 5.9 Hz), 3.94 (1H, t, J = 6.2 Hz), 4.33 (lH, d, J = 5.5 Hz). MS found 330.0 [M + H] +, 351.9 [M + Na] +. HRMS estimates C13H19N307, 330.1301. Found: 330.1295. Example 30 nSJSJS.SRjRU-CSW-diamino-hexylamino) -4-hydroxy-ditan 1.01Hexionate> -2,6-diboronic acid dihydrochloride-160- 432 S5435 200400815

Prepared according to general step C, using (1S, 2S, 4S, 5R, 6R) -2- (2 $, 6'-bis-third-butoxycarbonylamino-hexamidineamino) -4-hydroxy-bicyclo [3.1.0] Hexane-2,6-dicarboxylic acid diethyl ester (Preparation 31). ] n NMR (400 MHz, CD3OD) δ 1.47-1.59 (3Η, m), 1.66-1.78 (2Η, m), 1.84 (1H, dd, J = 5.4, 15.2 Hz), 1.91 (2H, m), 2.16 (1H, dd, J = 3.4, 5.7 Hz), 2.45 (1H, d, J = 15.7 Hz), 2.66 (1H, dd, J = 2.9, 5.9 Hz), 2.97 (2H, t, J = 7.3 Hz) , 3.92 (1H, t, J = 6.4 Hz), 4.33 (1H, d, J = 5.4 Hz). MS found 330.0 [M + H] +. Example 3 1 (1 8,28,48,51610-2- (2,8 -Yueanji 31 -Yuean methyl breweryl-propionyl sulfonyl) -4 and base-bicyclic "3.1.01 Burn -2,6 -diboronic acid

The title compound was prepared with (1S, 2S, 4S, 5R, 6R) -2- [2i-Third-Butoxy-161-4 85435 200400815 Hexylamino · 3, _ (triphenylfyl_amine methyl alcohol) ) _Propanylamino] 4_protecting group-difluorene [3.1.0] Hexyl U, 6-monocarboxybisbisethyl diethyl ester (0.77 mole, Preparation 32) is dissolved in THF (12 hairs) L), and the solution was treated with 2.5 N LiOH (12 ml). The solution was disturbed for 3 hours and then adjusted to pH = 2 with 2.5NΗα. The product was extracted four times with ethyl acetate, dried over anhydrous sodium sulfate and concentrated under vacuum to give a white powder. The diacid product (1.22 mmol) was dissolved in U2_dichloroethane (6 ml), and then treated with anisole (1 ml) and trifluoroacetic acid (6 ml). The reaction mixture was stirred at 23 ° C for 10 minutes. After hours, it was concentrated under vacuum to give a brown oil. The brown oil was diluted with diethyl ether and the product crystallized out as a white solid. Filter and wash with large: S 'ether. The solid was dissolved in a small amount of water, treated with 0.5 n HC 1 (5 ml) and lyophilized three times to give 386 mg (90%) of the title compound. 〇 Pull = 0 (c = 0.5, MeOH). ! H NMR (400 MHz, CD3〇D) δ 1.59 (1H, t, 3.4 Hz), 1.76 (1H, dd, J = 5.9, 16.1 Hz), 2.11 (1H, dd, J = 2.9, 5.9 Hz), 2.53 (1H, d, J = 16.1 Hz), 2.55 (1H, dd, J = 2.9, 5.9 Hz), 2.74 (1H, dd, J = 9.8, 17.1 Hz), 2.95 (1H, dd, J = 3.9, 17.1 Hz), 4.19 (1H, dd, J = 3.9, 9.8 Hz), 4.32 (1H, d, J = 5.9 Hz). MS found: 316.0 [M + H] +, 337.9 [M + Na] +. HRMS estimates CuHwNsC ^ Na, 338.0964. Found: 338.0953. Example 32 Π -3'-ΠΗ-Indolin-3-some V propionate 1-4-# empty-bicyclic "3.1.01 hexamethylene- 2,6-dipic acid 162- 4 34 85435

200400815 2- [2'-Third-butoxycarbonylamino_3, _ (1, _third-butoxycarbonyl_ih-pyridin-3, yl) -propanylamino] -4-hydroxy-di Diethyl cyclo [3_1.〇] hexane_2,6-dicarboxylate (0.93 mmol, Preparation 33) was dissolved in THF (1 ml) and the solution was treated with 2 5 N (11 ml). The solution was allowed to stir for 3 hours and then adjusted to pH = 2 with 2.5 NHC1. The product was extracted four times with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated under vacuum to give a white powder. The diacid product (0.78 mol) was dissolved in 4M HC1 in a solution of drank and burned (20 ml), and the reaction mixture was stirred at 23 for 3 hours. The reaction mixture was concentrated, diluted with dichloromethane and concentrated again under vacuum. The crude product was dissolved in methanol, applied to a radial chromatography (silica gel) plate, allowed to spin dry, and then MeOH (10%) / NH4OH (1%) / CHCl3 to MeOH (60%) / NH4OH (2%) / CHC13 dissolves. The product was dissolved in water and lyophilized twice to yield 139 mg (46%) of the title compound. [0 ^ = + 19.23 (c = 0.52, MeOH). * H NMR (400 MHz, CD3OD) δ 1-43 (1H, t, J = 2.9 Hz), 1.57 (1H, dd, J = 5.9, 15.2 Hz), 2.05 (1H, dd, J = 3.4, 6.4 Hz ), 2.41 (1H, d, J = 14.7 Hz), 2.46 (1H, dd, J = 2.4, 5.9 Hz), 3.15 (1H, dd, J = 5.4, 14.7 Hz), 3.49 (1H, dd, J = 5.4, 14.7 Hz), 4.05 (1H, dd, J = 5.4, 9.3 Hz), 4.17 (1H, d, J = 5.4 Hz), 7.09 (2H, m), 7.27 (1H, s), 85435 -163- 200400815 7.37 (1H, d, J = 8.3 Hz), 7.75 (1H, d, J = 7.8 Hz). MS found: 388.0 [M + H] +, 409.9 [M + Na] +. HRMS estimates C19H21N306, 388.1508. Found: 3881.502. Example 33 Jiang S, 2US, 5R, 6R) -1-several bases than diandian-2'S-carbonyl V neck certain μ bicyclo hexane-2,6-di #

Prepared according to general step c, using (1S, 2S, 4S, 5R, 6R) _2_ (1, _third butoxycarbonyl-pyrrolidine-2'S-carbonyl) _4_hydroxy_bicyclo [31〇] hexane_2 Diethyl 6,6-dicarboxylate (Preparation 34), except for removal of the third butoxycarbonyl protecting group, was treated with 4 M HC1 / dioxane. [〇 ^ 3 = -40 (c = 0.5, MeOH). 'H NMR (400 MHz, CD3OD) δ 1.59 (1H, t, J = 3 Hz), 1.78 (1H, dd, J-5.9, 15 · 3 Hz), 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). HRMS estimates C13H19N206, 299.1243. Found: 299.1242. Analysis estimates c, 44 26; H, 600; N, 7%. Found: C, 44.13; H, 5.78; N, 7.62. Example 34 8,4 511,611) 11 ^ 2,5-Amine-3,-(4-hydroxysome-benzyl, _1 ^ Saye ^ -164-4; ^; ρ 85435

OH

Prepared according to general step C, using (1S, 2S, 4S, 5R, 6R) -2- [2, S- $ st oxocarbonylamino-3,-(4-tert-butoxycarbonyloxy_phenyl) _Propylamido] -4_hydroxy-mono- [3.1.〇] hexane_2,6-dicarboxylic acid diethyl ester (Preparation 35), except for the third butoxycarbonyl protecting group using 4 M HC1 / difluorene Remove with alkane treatment. Yield: 199 mg (50%). [° C = + 8 (c = 0.5, MeOH). 'H NMR (400 MHz, CD3OD) δ 1.48 (1H, t, J = 3 Hz), 1.62 (1H, dd, J = 5.9, 15.3 Hz), 2.03 (1H, m), 2.38 (1H, d, J = 15.3 Hz), 2-55 (1H, m), 2.8 (1H, dd, J = 8.9, 14.4 Hz), 3.12 (1H, dd, J = 5.4, 14.4 Hz), 3.90 (1H, q, J- 5.4 Hz), 4.21 (1H, d, J = 5.9 Hz), 6.7 (2H, d, J = 8.4 Hz), 7.05 (2H, d, J = 8.4 Hz). HRMS estimates C17H21N2O7, 365.1349. Found: 365.1374. Knife analysis estimates C17H20N207 * L1 HCl-1.1 H20: C, 48.12; Η, 5.54. N 6.6. Found: C, 47.89; H, 5.37; N, 6.50. Example 35 LH2_§_r4S, 5R, 6R) -2- (2’S-amino-4'-methylsulfanyl-butyl hydrazone | amine group) _4_ invite -165-85435 200400815

Prepared according to step C, using (1S, 2S, 4S, 5R, 6R) -2- (2, S4ST oxygen Ik-based amino-4 -methylthio_butanylamino) _4 · hydroxy-bicyclo [ 3 · 丨 .〇] Hexane-2,6-- · dicarboxylic acid diethyl ester (Preparation%), except that the third butoxycarbonyl protecting group was removed by treatment with 4MHC1 / dioxane. The final compound was added to the 011 plate with ^^ 〇11 and 2 valence 011 * 〇11 ^ price, and was 50 / 49.5 / 0.5.11 (: 13 / Me〇H / NH4〇H Dissociation. Throughout the dissociation process, the amount of NΗ4〇η was increased to 1% to form a zwitterionic compound. Yield 136 mg (37%). [Α] Lan = + 28 (c = 1.0, MeOH).! H NMR (400 MHz, CD3OD) δ 1.63 (1H, dd, J = 6.0, 15.6 Hz), 2.03 (2H, m), 2.12 (3H, s), 2.18 (1H, m), 2.35 (1H, m), 2.49 (1H, d, J = 15.6 Hz), 2.65 (2H, t, J = 8.4 Hz), 3.84 (1H, t, J = 6.0 Hz), 4.19 (1H, d, J = 6.0 Hz). HRMS estimate C13H21N206S , 333.1120. Found: 333.1105. Example 36 (15:!? 48,511,6 Phen-2- "2 | 8- (28-Amine-Propyl: Amine.Base) -Propyl-Neck &amp; 1- 4-Hydroxy-bicyclo Π. 1.01 Hexane-2.6- Dimiaoling 85435 -166-

200400815 (1S, 2S, 4S, 5R, 6R) -2- [2'S- (2S-Third-butoxycarbonylamino-propanamido) -propanamino] -4-hydroxy-bicyclo [3.1. 0] Hexane-2,6-dicarboxylic acid diethyl ester (0.045 g '0.1¾ Mor, Preparation 39) was stirred and mixed in 1 ml of 4 NHC1 / Nisaki for 40 minutes, and then concentrated under vacuum. The crude material was stirred in THf (5 ml) and 0.5 N LiOH (0.35 mmol) for 1 hour. Adjust to ρΗ = 2 with 0.5 N HC1 and concentrate the reaction. The crude product was isolated by ion exchange chromatography (Dowex® 50X8-100: 10% pyridine / H2O) to give 19 mg (55.3%) of the title compound. lU NMR (300 MHz, D20) δ 1.23 (3Η, d, J = 7.3 Hz), 1.36 (3H, d, J = 7.3 Hz), 1.40-1.45 (1H, m), 1.50 (1H, dd, J = 5.5, 15.4 Hz), 1.90-1.96 (1H, m), 2.20 (1H, dd, J = 2.6, 5.9 Hz), 2.29 (1H, d, J = 15.4 Hz), 3.90 (1H, q, J = 7.3 Hz), 4.15-4.22 (2H, m). MS (ES) m / z 342.1 [M-1]. HRMS estimates C14H22N307, 344.1458. Found: 344.1457. Example 37 (18'28, 48, 51 ^, 61〇-2- "2 | 8- (2- || Anji-Yi donkey Yueshen)-propyl hydrazine Anji 1_4-Baji-II? "3.1.01 Hekeng-2,6 -dijinic acid

200400815 (13,23,48,511,6) -2- [2'8- (2-Third-butoxycarbonylamino_acetamido) -propanylamino] -4-epi-bicyclo [ 3.1.0] Diethyl hexamethylene-2,6-dipropionate (0.12 g '0.25 mmol, Preparation 40) in THF (5 mL) and 1 N LiOH (1.0 mmol) for 1 hour. The pH was adjusted to pH = 2 with 1 N HC1 and concentrated. The crude material was stirred in a saturated ethyl acetate solution of HC1 (gas) at 0 ° C. Filled with &amp; to remove excess HC1 (gas) and concentrated Reaction. The crude product was purified by cation exchange chromatography (Dowex® 50X8-100; dissolved at 10% Edo / h20) to yield 0.06 g (72.9%) of the title compound. [Αβ3 = -42.11 (c = 0.57, H20). 4 NMR (300 MHz, D20) δ 1.21 (3H, d, J = 7.3 Hz), 1.39 (1H, 'm), 1.50 (1H, dd, J = 5.7, 15.8 Hz), 1.91 (1H, m), 2.15 (1H, dd, Lu J = 2.6, 5.9 Hz), 2.28 (1H, d, J = 15.0 Hz), 3.65 (2H, s), 4.14 (1H, app d, J = 5.9 Hz), 4.20 (1H, app q, J = 7.3 Hz). HRMS estimates C13H2ON307, 330.1301. Found: 330.1299. _ Example 38 nS, 2S, 4S, 5R, 6R) -2- "2'S- (2S-amino -4-A-Mou-Methylamine Air sign 1_4_ 衮 Mou-bicyclic 「3.1.01 Hexane-2.6-Di-taebo 85435 -168-

Prepared as usual in step c, using (1S, 2S, 4S, 5R, 6R) _2_ [2, _ (2S_third butoxysynamino-4-amidino-pentylamino) _propanamido ] _4_Hydroxy-bicyclo200400815 D · 1.0] Diethyl 2,6 · dicarboxylate (Preparation 41). Yield 0.21 g (57%, 0.50 mmol) as a white solid. [a] 2D3 = -3.64. (C = 0.55, MeOH). lH KMR (300 MHz, CD3OD) δ 0.89 (3H, d, J = 4.4 Hz), 0.91 (3H, d, J = 4.0 Hz), 1.29 (3H, d, J = 7.0 Hz), 1.52 (1H , t, J = 3.3

Hz), 1.55-1.67 (4H, m), 1.97 (1H, dd, J: 2.9, 5.9 Hz), 2.38 (1H, dd, J = 2.9, 5.9 Hz), 2.45 (1H, d, J = 15.4 Hz ), 3.75-3.80 (1H, m), 4.18 (1H, d, J = 5.9 Hz), 4.33 (1H, app. Q, J = 7.0 Hz). Analysis estimates C17H27N3O7.1.0 HCU1.6 H20: C, 45.30; H, 6.98; N, 9.32; Cl, 7.87. Found: C, 44.95; H, 6.54; N, 9.12; Cl, 7.53. HRMS (ES) estimate Cl7H2SN3007 [M + H] +, 386.1927. Found: 386.1911. Example 39 (1S, 2S, 4S, 5R, 6R) -2-l ~ 2'S- (2-amino-3-methyl-butanylaminopropylsulfinoamino1-4-hydroxy-bicyclo Π. 1.01 hexane-2,6-di # age 85435 -169- 200400815

= Generally prepared in step c, using (1 s, 2s, 4s, 5R, 6R) _2_ [2, _ (2s_p Butylcarbamino_3_methyl-butyramido) -propanamido]- 4-hydroxy-bicyclo P.1.0] hexane-2,6-dicarboxylic acid diethyl ester (Preparation 42). Yield: 0.15 g (88%, 0.37 mmol) as a white solid. [«] 〇 = -15.69 ° (c = 〇5i? MeOH) 〇H NMR (300 MHz, CD3OD) δ 0.92 (3Η, d, J = 7.0 Hz), 0.95 (3H, d5 J = 7.0 Hz), 1.28 (3H, d, J = 7.0 Hz), 1.52 (1H, t, J = 2.9 Hz), 1.60 (1H, dd, J = 5.9, 15.4 Hz), 1.98 (1H, dd, J = 3.3, 5.9 Hz) , 2.03-2.15 (1H, m), 2.39 (1H, dd, J = 2.9, 6.2 Hz), 2.45 (1H, d, J = 15.4 Hz), 3.56 (1H, d, J = 5.5 Hz), 4.18 ( 1H, d, J = 5.9 Hz), 4.33 (1H, app. Q, J = 7.0 Hz), 8.82 (1H, s). Analysis estimates C16H25N3O7M.0 HCW1.5 H20: C, 44.19; H, 6.72; N, 9.66; Cl, 8.15. Found: C, 44.32; H, 6.48; N, 9.14; Cl, 7.66. HRMS (ES) estimates C16H26N307 [M + H] +, 372.1771. Found: 372.1758. Example 40 (1R, 2S, 4R, 5R, 6R) -Aminopropylammonyl) Amino-4 _ gas-di) anion 170- △ 42 85435 200400815 Two # acid formazan to one forever ..

Add 16 ml of water followed by 11.4 ml (175 mmol) of methanesulfonic acid to (111, 23,411, 5 feet, 611) -2- (2'8-2 '-(third butoxycarbonylamino) ) Propanyl) amino-4-fluoro-bicyclo [3.1.0] hexane_2,6-dicarboxylic acid (32.8 g, 87.5 mmol, Preparation 52) in a cloudy solution of 295 liters of acetone Medium, added dropwise under gentle reflux, a clear 'light yellow solution' can be formed, which forms a slurry after 5 minutes. Stir for 30 minutes, remove the heat source, and add 295 ml of acetone over 30 minutes. The slurry was cooled to room temperature and stirred for another 2 hours. The slurry was filtered, washed with acetone (2x82 mL) 'and dried under vacuum at 45 ° C for 16 hours to yield 31.5 g (93%) of the title compound as a white solid. The material may be further recrystallized. 30.5 g of the above crude product was mixed with 152.5 ml of acetone and 35 ml of water. Heat to 55 ° C and add water (3.66 ml) to achieve complete dissociation. The solution was diluted with 61 ml of acetone and seeded as needed. Remove the heat source 'and allow the mixture to cool gradually until good nucleation begins. To the slurry was added 396 ml of acetone over 70 minutes, and the mixture was stirred at room temperature for 3 hours. Filtration, washing with acetone (3x91 mL), and vacuum drying at 45 ° C for several hours (usually overnight) to yield 27.7 g (91% recovery) of the title compound as a white solid. mp (DSC) 200 ° C.

85435 -171 · 200400815 [a] 25D +34. (C 1.0, CH3OH). 400 MHz 'H NMR (DMSO-d6) 612.76 (br s, 2H), 9.18 (s, 1H), 8.07 (br s, 1H), 5.50-5.36 (m, 1H), 3.87 (d, 1H, J = 6.8 Hz), 2.82 (dd, 1H, J = 14, 8.0 Hz), 2.38 (s, 3H), 2.25 (m, 2H), 1.96 (t, 1H, J = 3.0 Hz), 1.39 (m, 1H) , 1.37 (d, 3H, J = 6.8 Hz). 100 MHz I3C NMR (DMSO-d6) δ 173.1, 172.3, 169.7, 92.8 (d, CF split), 63.2 (d, CF split), 48.0, 39.7, 36 · 4, (d, CF split), 3.25 (d , CF split), 29_3 (d, CF split), 19_3 (d, CF split), 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. Analyze and estimate C12H21FN209S: C, 37.11; H, 5.45; N, 7.2 Observed values: C, 37.12; 5., 5.45; N, 7.16 ° Example 41 L1R, 4S.5 8.68) -4- (25-4 ^ -A Sulfur-2'-amine, butylamine, amine-7. 9.-diketone-2λ6-4-bicyclo 丨 3.1.01 hexane-4,6-dicarboxylic acid-permanent makeup

Put (lR, 4S, 5S, 6S) -4- (2'S-4'_methylthioK tert-butoxycarbonyl) aminobutylfluorenyl) amino-2,2-diketo-2λ6-fluorene-bicyclo卩 .1.0] Hexane-4,6-di 4 4 85435 -172- 200400815 Monosodium carboxylic acid salt (110.04 g, 225.3 mmol, Preparation 55) in acetone (110 ml) and water (530 ml) The slurry was heated to 55t. Concentrated hydrochloric acid (56 ml ' 675.8 mmol) was added to the stirred slurry to gradually cause dissociation. — Once the complete Canadian dollar ’solution is dropped at 5 5 C for 2 hours’, remove the heat source and allow the solution to cool to room temperature. Rinse the solution and rinse with 20 ml of water. Slowly add 2 N sodium hydroxide (165 ml, 330 mmol) to the solution to raise the pH to 1.71, at which point precipitation occurs. After stirring for 10 minutes, a thin slurry formed and the pH dropped to 0.98. To the resulting slurry was added more 2 n sodium hydroxide (62 ml, 124 mmol) to raise the pH to 3.06, followed by stirring for 3 hours to produce a final pH of 3.24. The filtered slurry 'was washed with water (2x165 ml;) and dried at 45c for 15 hours to yield 73.27 g (85% by weight yield) of the title compound as a white solid. mp (DSC) 2030C. [a] 25D +13.4 (c 1.19, 1 N HC1). 500 MHz, 'H NMR (D20) δ 3.99 (t, 1H, J = 6.〇Hz), 3.93 (dj lH, J = 15.0 Hz), 3.50 (dd, 1H, J = l.〇, 4.0 Hz) , 3.12 (d, 1H, J = 15-〇

Hz), 2.95 (dd, 1H, J = 4.0, 7.0 Hz), 2.48 (t, 2H, J = 8.0 Hz), 2.33 (t, 1H, J = 4.0), 2.09-1.98 (m, 5H). 13C NMR (125 MHz, D20) δ 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), i748.83 (s), 1692.89 (s), 1681.99 (s), 1617.50 (s), 1567.63 (s); ^ 97.55 (s ), 1314.11 (s), 1282.22 (s), 1263.26 (s), 1230.01 (s), u0146 (s), 884.62 (s), 809.95 (s), 773.46 (s) cm-1. 85435 • 173- 200400815 Knife analysis estimate C12H18N207S2 * H20: C, 37.49; H, 5.24; N, 7.29. Found: C, 37.34; H, 5.04; N, 7.15. Example 42 Methylthio-2, Aminobutanine) Amine-2,2-dihydro-,-2,-, cyclic ring "3.1.01 have been married. -4,6-ditransformic acid formate

HOjC

Put (lR, 4S, 5S, 6S) -4- (2'S-4'_methylthio-2,-(third butoxycarbonyl) aminobutylfluorenyl) amino-2,2-diketo-2λ6- Pyrene-bicyclo [3.1.0] hexane-4,6-dicarboxylic acid (118.09 g corrected, 253 mol, preparation 56) p-toluenesulfonic acid monohydrate (54 g, 278 mmol) ) The mixture in toluene (1180 ml) was heated to 75 ° C. This resulted in a thick slurry. The slurry was stirred at reflux for 165 minutes. The heat source was removed and the slurry was allowed to cool to room temperature and stirred overnight. Filter the slurry: 'Wash with toluene (3 x 240 ml) and then at 45. (: Drying under vacuum for 22 hours to yield 134.92 g (98% yield) of the title compound. Mp (DSC) 255 ° C. [A] 25D +8.3 (c 1.2, CH3OH). 500 MHz, NMR (CD3OD) 67.71 (d, 2H, J = 8.0 Hz), 7.24 (d, 2H, J = 8.0 Hz), 4.14 (d, 1H, J = l5 Hz), 4.00 (t, 1H, J = 6.0 Hz), 3.54 (dd , 1H, J = 4.0, 7.0 Hz), 3.13 (d, 1H, J = 15 Hz), 3.01 (dd, 1H, J = 4.0, 7.0 Hz), 2.60 (t, 2H, J = 8.0 Hz) , 2.49 (t, 1H, J = 4.〇446 85435 -174 &lt; 200400815

Hz), 2.37 (s, 3H), 2.19-2.12 (m, 5H). 13C NMR (125 MHz, CD3OD) 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 ". Example 43 m4US, 6S) -4- (2'S-4'-methylthio- 21-Amine butyl _ even) amine_2,2-di 6-Γ3.1 · 〇1 hexane-4,6-di # _

Dibutoxycarbonyl) amino

‘丨 Soil to ㈤, stir for another minute. I ’ve dried it for 4 nights. Through hydrazone, 1.04 g (97%) of (1R, 4S, 5S, 6S) -4- (2'S-4'-methylthio_2 '-(third butyl fluorenyl) amino) -2,2- Diketonic acid (1.08 g, 2.31 mmol) was heated to 85 ° C. Water (540 徜 microliters, 3.47 mmol) was added. The slurry was stirred for 90 minutes. Nitrile (30 mL). The slurry was cooled to room temperature Washed with M surname c Nitrile (3 \ 2.7 ml) 'at 45. (: dried overnight, may be the title compound. 85435 -175- 200400815 mp (DSC) 244 ° C. [A] 25D +10.2 (c 1.16, CH3OH). 500 MHz,] H NMR (CD3OD) δ 4.16 (d, 1H, J = 15 Hz), 4.00 (t, 1H, J = 6.0 Hz), 3.54 (dd, 1H, J = 4.0, 7.0 Hz) , 3.15 (d, 1H, J = 15 Hz), 3.01 (dd, 1H, J = 4.0, 7.0 Hz), 2.71 (s, 3H), 2.61 (t, 2H, J = 8.0 Hz), 2.51 (t, 1H, J = 4.0), 2.20-2.14 (m, 5H). 13C NMR (125 MHz, CD3OD) δ 170.50, 169.71, 169.〇〇, 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), ΛΓ 1528.96 (s), 1320.89 (s), 1176.86 (s), 1152.70 (s), 1118.55 (s), 1051.42 (s), 816.49 (s), 786.63 (s) Points-1. Analysis and estimation C12H18N207S2.CH403S: C, 33.76; H, 4.79; N, 6.06. Found: C, 33.98; H, 4.82; N, 5.98. Case 44 (111,23,511.610-2- (2 | 11-neck, even 醯）) amine -6- exhausted-di 秾 『1 ^^ hexane-2,6-di # hydrochloride. —Win

Canton 0 NH / Cr

Me, &quot; 1.13 ml (2.8 mmol) of 2.5 N LiOH was added to 0.20 g (0.47 mmol) of 60 isomer A in a solution of THF (1.2 ml), and the resulting mixture was placed in a chamber. Stir overnight at warm temperatures. Wash with ether 'and neutralize with 1 N HC1 at 0 ° C, then extract with EtOAc (3x3 mL). The combined organic layers were dried over anhydrous MgS04 85435 -176- 200400815 and concentrated under reduced pressure. The crude was dissolved in 3 76 ml of 1 n HCl / EtOAc and stirred overnight. The solvent was decanted and washed with diethyl ether.

Ar-steam drying gave the title compound (83 mg, 57% yield) as a white solid. H NMR (300 MHz, CDC13): 1.28 (d, 3H, J = 7.1 Hz), 1.64-1.69 (m, 1H), 1.86-2.02 (m, 1H), 2.11-2.21 (m, 3H), 2.54 ( d, 1H, J = 7.1 Hz), 3.84 (q, 1H, J = 7.1 Hz). Example 45 ilHP ~ '5S, 6S) -2- (2-Di-R-amino-anisopropyl) amine -6-excited diamine "3 丄 〇1hexane 毽 -.2,6-; carboxylic acid salt

NH3 + C |. The solution of isomer B from Preparation 60 &apos; can be prepared essentially as described in Example 44 to the title compound. ! H NMR (300 MHz, CDC13): 1.30 (d, 3H, J = 7.1 Hz), 1.59 ^ (m, 1H), 1.85-2.03 (m, 1H), 2.05-2.25 (m, 3H), 2.53 ( d, m J = 6.6 Hz), 3.85 (q, 1H, J = 7.1 Hz). '' Example 46 (1S, 2S, 4S.5R, 6R) -2- (2'S --- aminopropylammonium) amine · 4_ Kaojidi Γ3 · 1.01 ^ 2,6-dicarboxylic acid hydrochloride 85435- 177 ·

200400815 Prepared according to the general procedure C, using (1S, 2S, 4S, 5R, 6R) -2- (2, S-fST oxoamino-propionylamino) -4-hydroxy-bicyclo [3.1 .0] Hexane-2,6-dicarboxymethyl-ethyl acetate (0.47 g 'mmol, preparation 37) yielded 0.23 g (67.7%) of the title compound. [<U (c = 0.49, MeOH). H NMR (CD3OD) δ 1.51 (3H, d, J = 7.0 Hz), 1.56 (1H, t5 1 = 2.9 Hz), 1.77 (1H, dd, J = 5.8, 15.8 Hz), 2.1-2.13 (1H, m ), 2.52 (1H, d, J = 16.1 Hz), 2.57 (1H, dd, J = 2.9, 5.9 Hz), 3.88 (1H, dd, J = 7.〇, 14.3 Hz), 4.32 (1H, d, J = 5.5 Hz). Analyze HCU0.9 H20: C, 40.12; H, 5.80; N, 8.53. Found: c, 39.85; H, 5.41; N, 8.36. HRMS estimated CuH16N206Na: 295.0906. Found 295.0883. The prodrug compound of the present invention can be evaluated by various cell uptake analysis methods corresponding to the parent compound. The comparative data provided by these assays allows the artist to identify compounds that are indeed absorbed into cells and provide excellent exposure. The two analysis methods are: Gly-Sar intake analysis method and cac02 analysis method, which will be described in detail below.

Glv-Sar Intake Analysis It is known that the absorption of certain oral curdrin drugs is through the enteral delivery system. Yang, et al. Pham. Res. 16 (9) (1999). In particular, the intestinal peptide transporter (hPepTl) has been studied for the inhibitory performance of skin intake 85435 -178- 200400815 and its considerable level confirmed in cells. Meredith, et al., Eur. J. Biochem., 267, 3723-3728 (2000). Furthermore, identifying the intestinal absorption mechanism of amino acids in the hPepTl transporter has become the target of identifying effective strategies to improve oral absorption by oral administration. Han, et al., Polym.

Prepr. (Am. Chem. Soc., Div. Polym. (Chem) 40 (1): 259-260 (1999); Sawada, et al. J Pharmacol. Exp. Ther., 291 (2): 705 -709 (1999). US Pat. No. 5,849,525 describes a method that can be used to measure the compounds of the present invention: the level of affinity for hPepTl transporter. For example, the stably transfected Chinese hamster egg of hPepT 1 transporter ( CHO) The fine month pack 'Yes; to test the compound of the present invention. The absorption of Gly-Sai · in the cell can be tracked' When absorbed in the presence of the prodrug compound of the present invention, its 1st and 1th classic compound of the present invention This may be an indication of agonist activity; and when several people of the prodrug of the invention &amp; a-Gramineous compounds usually exhibit values of less than 5mMi · EC50.

Caco-2 analysis Another special method for measuring the ingestion of this meal compound into cells is to study the peptide transporter of human intestinal cell line CaC0_2. Human adenocarcinoma cells (Memorial Sloan-Kettering Can ~ ^ ‘ncer Center, Rye, NY, and / or ATCC,

Rockville, MD) in Dull · », ΛίΓ,

Passaged in Uibecco s Modified Eagle medium, which contains 10% fetal bovine blood, fen · ι 0 / say ** Mo Yi 丄 β and 1 / 〇 take non-essential amino acid solution with low nutrition requirements, not Add C-age H-Master, two t-nano or antibiotics. These cells were mycelium-free and 85435 -179- 200400815 was used at 28 and 40 passages. For flow measurement, 5 to 10x104® cell cultures are performed in collagen-coated porous hole cultures for 13-18 days, and the medium is changed every 2 to 3 days. Drug ingestion was performed at 37 ° C using a cluster-tray technique with test compounds. (See 0 冱 22〇1, 61 冱 1., 11 &amp; 1 · Biochem. 1, 15, 368-74 (1981)). The flowing buffer solution was Earle's balanced salt solution without bicarbonate, which contained 25 mM Mes, adjusted to pH 6.0 with KOH, and replaced sodium chloride with choline chloride test. The permeability of the mobile buffer solution was adjusted to 300 ± 5 mosmol / kg with the help of choline chloride test. [3H] Inulin is used as a standard for extracellular fluids. It can adhere to the cells during the washing process to estimate the zero hour and determine the intake rate. Fresh solutions of test compounds and dipeptides are prepared daily. At the end of the experiment, the cells are lysed in water, and the test compounds can be detected in the lysate by LC / MS / MS. Proteins were measured using the method described in Smith, et al., Anal. Biochem. 150, 76-85 (1985). Intake was measured over 40 minutes. The initial intake rate is calculated in the linear region of the time course regression, and the estimated zero time uses linear regression as described above. The inhibition rate was calculated based on the intake rate of the control group measured in the absence of dipeptide. For this Caco-2 analysis, see Dantzig &amp; Bergin, Biochem. Biophys. Acta 1027, 211-17 (1990). Determination of in vivo exposure using rat plasma concentrations To study the in vivo exposure of a compound of formula II to a compound of formula II after oral administration of a compound of formula I, the plasma concentration of individual compounds of formula II was measured in rats. Mature Fischer 344 male rats (190-270 g) were obtained from Harlan 85435 -180- 200400815

Sprague-Dawley, Cumberland, IN, USA, and acclimated in the laboratory for 3 days. On day 4 'the test compound was dissolved in buffered water (1 mg / ml = test compound / 20 mM potassium dihydrogen phosphate, pfj = 2), and was administered orally at a single dose of 5 mg / kg. Blood samples were collected at 0.5 and 1 hour, or 1 and 3 hours via the eye sinus or heart (last time point). The plasma sample was stored at -20 ° C and analyzed in the presence of phenylsulfenylsulfonium fluoride. This compound is a protease inhibitor. Plasma samples and internal standard compounds were pretreated by solid phase extraction (SAX carrier, methanol / water / dilute acetic acid as shown in Tables 1A and 1B below. LC / MS / MS was used to determine the plasma of individual compounds of formula II for each test compound Concentration (nanograms per milliliter), and expressed as the sum of concentrations at 0.5 and 1 hour, or another 1 and 3 hours of sampling | ----- / Eigh / person ------- --- 1 A1A in vivo exposure of compound compounds in rats [nanograms / ml of (111,48,58,63) -4- (2 &amp; aminopropylamido) amino-]-2,2- Diisoamyl-2, 6-1? -Di-? A [3,1.0] (see -4,6-—carboxylic acid hydrochloride) Example 1 2251 ng / ml (10 mg / kg) After P · 〇.) The non-prodrug version of Example 1 is 1521 ng / ml (5 mg / kg P.0 ·) 3981 ng / ml (10 mg / kg P.0. 85) 85435 Table 1B in vivo Exposure Analysis Compounds Rat exposure (nanograms / ml of (1 R, 2S, 4R, 5R, 6R) -2- (2'S-amino-propanyl) amino-4-fluorobicyclo [3.1.0] Hexane-2,6-dicarboxylic acid hydrochloride) Example 14 5271 ng / ml (after 5 mg / kg po) Non-prodrug type of Example 14 Formula 1162 ng / ml (after 5 mg / kg ρ · ο ·) 1342 ng / ml (after 10 mg / kg ρ · ο.) 200400815 As shown in Tables 1A and 1B above, when administered orally to rats, the present invention The compounds exhibit comparable plasma concentrations to the parent compound. This demonstrates that the compounds of the invention can be converted into the parent compound in vivo, i.e. compounds of formula II. The compounds of the invention are preferably formulated before administration. Therefore, another aspect of the present invention is a pharmaceutical blend comprising a compound of formula I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient. Pharmaceutical blends are prepared using procedures well known to those skilled in the art. In preparing the composition of the present invention, the active ingredients are usually mixed with a carrier, diluted by the carrier, or enclosed in the carrier, and may be in the form of capsules, foamable granules, paper or other containers. When the carrier is used as a diluent, it can be a solid, semi-solid or liquid substance, and can be used as a solvent, excipient or medium for the active ingredient. The composition can be in the form of tablets, pills, powders, sugar bonds, foamable granules, axons, suspensions, emulsions, solutions, syrups, aerosols, ointments, containing up to 10% by weight Active ingredients, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders. Examples of suitable carriers, excipients, and diluents include: lactose, dextrose, 85435 -182- 200400815 sucrose, sorbitol, mannitol, starch, gum 'acacia, calcium phosphate, alginate, scutellaria baicalensis Gum, gelatin, calcium silicate 'microcrystalline cellulose, polyvinylpyrrolidone' cellulose, syrup water, hydrazone, methyl hydroxybenzoate and propyl ester 'talc, stearic acid to ease mineral oil. Mixtures can include lubricants, moisturizers ... or fragrances as required. The composition of the present invention can be adjusted so that the active ingredient can provide rapid, continuous or delayed release after being administered to a patient through steps known in the art. The composition is preferably blended into a unit dosage form, each dose containing from about 5 mg to about 500 mg of the active ingredient ', preferably from about 25 mg to about 300 mg of the active ingredient. As used herein, &quot; active component &quot; refers to a compound within the range. a ', unit dosage form, refers to physically separated units, suitable for humans and other mammals &lt; early dose' each unit contains a predetermined amount of active ingredients, plus: a combined pharmaceutical carrier, diluent Or tincture, which produces the desired healing as 85435 -183 «

Claims (1)

200400815 Patent application park: 1. A compound of formula I Where A is H- (Q) P-; Q is independently selected each time from aminoamino; p is an integer from 1 to 10; X is 0, S, SO, S02 or CR3R4; R3 is fluorine, X ' OR5, S03H, tetrazol-5-yl, CN, P03R62, hydroxyl, No2, N3, (CH2) mCOOR5a, (CH2) mP03R6a2, NHC0NHR5b, or NHS02R5e, and R4 is 氲; or R3 and R4 each represent fluorine ; Or R3 and R4 — represents = 0, = NOR7, = CR8R9, = CHCOOR5b, = CHP03R6a2 * = CHCN; or one of R3 or R4 represents an amine group and the other represents a carboxyl group; X 'represents a bond, CH2 or C0; m is an integer from 1 to 3; R5, R5a, R5b, R5e, R7, R8 and R9 are independent hydrogen atoms; (1-6C) alkyl is substituted as required; (2) is substituted as required -6C) alkenyl; (2-6C) alkynyl substituted as required; aromatic group substituted as required; heteroaromatic group substituted 85435 200400815 as required; non-aromatic carbocyclic group; non- Aromatic heterocyclic groups; non-aromatic early ring carbocyclic groups, which are fused with one or two monocyclic aromatic or heteroaromatic sandwiches; or non-aromatic monocyclic heterocyclic groups, which are related to One or two monocyclic ^ or heteroaromatic groups are fused; R6 and R6a are independent Represents hydrogen or (1-6C) alkyl; R1g is hydrogen or fluoro; Cloth hydrogen and R 'fluoro or several group; or a pharmaceutically acceptable salt thereof. 2_ The compound or its salt according to item 丨 of the scope of the patent application, the limitation is that the compound or its salt is not parent: (: 113 feet 4, where R3 is fluorine and R4 is hydrogen, p is 1, and Q is L -Alanine group; or a pharmaceutically acceptable salt thereof. 3. A compound or salt according to item 1 or 2 of the scope of patent application, wherein A is H- (Q) p-; Q is independently selected from amine group each time Fluorenyl; P is an integer from 1 to 3; x is 0, S, so, so2 or CR3R4; r3 is fluorine or hydroxyl, and R4 is hydrogen; or R3 and r4 together represent = 0; R1Q is hydrogen or fluorine And R is hydrogen 'fluorine or a side group. • A compound or salt according to any one of items 1 to 3 of the scope of the patent application, wherein Q is an aminofluorenyl group derived from a natural amino acid. 5' According to the patent application A compound or salt according to any one of the items 1 to 4 in which x is s0 2., • A compound or salt according to any one of the items 1 to 4 in the patent application, wherein 200400815 X is CR3R4 and R3 is Fluorine and R4 is hydrogen. 7 'A compound or salt according to any one of claims 1 to 4 of the scope of the patent application, wherein X is CR3R4, R3 is hydroxyl and R4 is hydrogen. 8. According to the application The pharmaceutically acceptable salt of item 1 of the patent scope is an acid-addition salt made with an acid, which can provide a pharmaceutically acceptable anion; and a tested base-addition salt, which can provide Compounds containing an acidic moiety are medically acceptable anions; or zwitterionic compounds containing an oppositely charged group. 9 'Compounds according to item 1 of the scope of patent application, where A is H- (Q) p-; Q is L -Propylamine; P is 1; X is so2 or CR3R4; R3 is fluorine and R4 is hydrogen; R1G is hydrogen; and R11 is hydrogen; or its hydrochloride salt, toluate, formate, ethyl Acid salt, sulfonate, or monosodium salt. 10. The pharmaceutically acceptable salt according to item 9 of the scope of patent application, which is (^ ... "^"-( The meaning of diketo-Zhong 6-chuan-bicyclo [3.1.0] hexane-4,6-dicarboxylic acid hydrochloride or (ir, 4S, 5S, 6S) -4_ (TS-2'- Aminopropylamido) amino-2,2_diketo-2λ6-pyrimidine-bicyclo [3.1.〇] Hexane-4,6-dicarboxylic acid tosylate. I According to the scope of application for patent i Compounds of the formula (1R, 4S, 5S, 6S) _ regrets 8543s 12 2004 00815 4- (2'S-4'-methylthio_2, _aminobutyridinyl) amino] _2,2_diketo_2λ6_thia-bicyclo [3.1 .〇] hexane_4,6_di Carboxylic acid or its pharmaceutically acceptable salt. • The compound according to item 11 of the patent application park, which is (1R, 4S, 5S, 6S &gt; 4_ (2'S-4'-methylthio · 2, _aminobutyridinyl) ) Amino-2,2-diketo-2λ6-4_ monocyclic [3.1.0] hexane_4,6-dicarboxylic acid monohydrate. 1. A pharmaceutically acceptable salt according to item 1 of the scope of application for a patent, which is 1S, 2S, 4S'5R, 6R-2- (2'S-aminopropylamino) amino_4-quinyl-bicyclo [ 3.1.0] Hexane-2,6-dicarboxylic acid hydrochloride. 14. The compound according to item i of the scope of patent application, wherein A is H- (Q) p-; Q is L-propylaminofluorenyl; P is 1; X is CR3R4; R3 is fluorine and R4 is hydrogen; r1g is hydrogen And R11 is hydrogen; or a pharmaceutically acceptable salt thereof. 15. The compound or salt according to item 14 of the scope of patent application, which is selected from the group consisting of: &amp;) 111'28,411,511,611-2- (2'8-2'-aminopropylamido ) Amino_4-fluorobicyclo [3.1.0] hexane-2,6-dicarboxylic acid hydrochloride; b) 1 foot, 28,4! 1,511,611_2_ (2'3_2 | _amino group Propionyl) amino-4_fluorobicyclo [3.1.0] hexane_2,6-dicarboxylic acid methanesulfonate; c) 111,28,411,5 feet, 611.2 (2'8_2, _ Aminopropylamido) amino-4_fluorodi-4- 459 85435 200400815 ring [3 丄 〇] hexane-2,6-dicarboxylic acid ethanesulfonate; d) 1 ^, 2S, 4R, 5R ， 6R_2_ (2, S_2,4-based propylamine-based bicyclo [3.1.0] Hexane_2,6_dipropionate benzoate; e) ^, 2S, 4R, 5R, 6R_2_ (2 , S_2mM) M_4j Bicyclo [3.1.0] hexane-2,6-dicarboxylic acid tosylate; f) 1R, 2S, 4R, 5R, 6R'2- (2, s-2mMm &amp; -4- b ring [3.1.0] hexane_2,6-dicarboxylic acid; and g) 11 ^, from the 2- (2,8-2, -aminopropylamido) amino_4_fluoro [3.1_0] Hexamidine-2,6-dicarboxylic acid monosodium salt. 16. The pharmaceutically acceptable salt according to item 15 of the patented Fr. patent park, which is (1 &amp; '28'411'511'61 ^ 2- (2 | 8-2'-aminopropylamido) amine -4-Fluoro_bicyclo [3.1_0] Hexane-2,6-dicicarboxylic acid mesylate. 17. A pharmaceutically acceptable salt according to item 16 of the scope of patent application, which is (1R, 2S , 4R, 5R, 6R) -2- (2, S-2, -aminopropylamido) amino-4-fluoro.Bicyclo [3.1,0] hexane-2,6-dicarboxylic acid methanesulfonate Acid salt monohydrate 18. A method of preparing a compound of formula j or a pharmaceutically acceptable salt thereof as claimed in item i of the scope of patent application 'This method comprises the halogenation of a compound of formula (ii) Use the corresponding amine fluorenyl group PgN-A- 85435 (III) 200400815 of formula III 19. where PgH is a nitrogen-protecting group; "for any of the above steps," when using a protective group to protect a functional group, remove the protective group; after , For any step of the above; when the formula is required to be a pharmaceutically acceptable salt of the compound, the age of the compound of the formula I is reacted with an acid that can generate a pharmaceutically acceptable counter ion. ΛM &amp; 土 ' Or for compounds with the acidic part of the formula. 1 'React this formula with a base, fly the sigma-sense acid type, which can generate medically ignitable cations; or needle plate 4 T it,, the formula I zwitterionic compounds, then neutralize the acid addition salt of the compound of formula I and test A a _ on the fly addition salt; or go through any other conventional steps. A method that can affect the patient's CAMP-follow-up, and then stimulate the metabolic glutamate receptor =, this method includes the need to regulate the stimulating amino acid neurotransmission = the basis for administering a pharmaceutically effective dose A y * 2 0 of the scope of application for a patent application. A method for administering an effective dose of a compound of formula II H0, c R 丨 fl ',,, -X (Π) 21. Where x and Rl0 are as defined in the first patent application scope. This method includes administering Φ. ^ &Amp; cardiac stimulating amino acid neurotransmission for patients who need to be adjusted, and administering the medicine to the patient. Effective dose of the root compound. The application is called the first item of osteomyelitis in the treatment of patients with neurological disorders, and this method includes administering a medically effective dose of a compound according to item 1 of the scope of the patent to a patient who needs the treatment of 85435 2004008 ^ 5. . The method according to item 21 of Shengu's patent scope, wherein the neurological disorder is a brain defect after graft-to-visceral circuit surgery and transplantation; cerebral hemorrhage; lunar column trauma, head shao trauma; Azheimer's Disease; Huntington's disease; amyotrophic lateral sclerosis; dementia by aids-defamation; hypoxia during production; hypoglycemic neuron damage; eye injury and retinopathy; cardiac dysfunction; Primary and Drugs_Study on Parkinson's disease, muscle cramps; migraine; urinary incontinence; drug resistance; withdrawal; discontinuation and cancer; smoking cessation; vomiting; cerebral edema; long-term pain; sleep disorders; Convulsions; Tourette's syndrome (Tourette, syndromo); attention deficit disorder and bradykinesia. 23 · «Method 22 of the scope of patent application, wherein the neurological disorder drug resistance, withdrawal, discontinuation and addiction; or quit smoking. 24. A method for treating a patient with a psychiatric disorder, the method comprising: 2Therapeutic patients vote? For the basis of a pharmaceutically effective dose, please patent it! The compound of item 1. 25. The method according to item 24 of the patent application, wherein the psychiatric disorder Schizophrenia, anxiety and related disorders, depression, bipolar disorders: mental illness and obsessive-compulsive psychosis. 26. The method according to item 25 of the scope of patent application, its anxiety and related disorders. 〃Zhongshen disease disorders: 27 · —A kind of medical concoction, which contains a magic compound, or a pharmaceutically acceptable green, plus a pharmaceutically acceptable carrier ’diluent or excipient. 85435 200400815 柒. Designated representative map: (1) The designated representative map in this case is: (). (2) Brief description of the element representative symbols of this representative figure: 捌 If there is a chemical formula in this case, please disclose the chemical formula that can best show the characteristics of the invention 85435