TW200407310A - Compounds - Google Patents

Abstract

The invention relates to compounds of formula (I), wherein R1 represents C1-6 alkyl; R2 represents pyrazole or pyrimidine; or a pharmaceutically acceptable derivative thereof. The invention also related to pharmaceutical compositions containing compounds of formula (I) and to the use of compounds of formula (I) in medicine, particularly in the amelioration of a clinical condition for which a ACE and/or NEP inhibitor is indicated.

Description

200407310 10 15 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 20 A7 B7 V. Description of the invention (1) Field of the invention The present invention relates to thiol derivatives with metalloproteinase activity, and more particularly mixed ACE-NEP inhibitory activity N-mercaptofluorenylphenylalanine derivative; a pharmaceutical composition containing the same and its use in medicine. BACKGROUND OF THE INVENTION Angiotensin-converting enzymes (ACE) and neutral endopeptidases (NEP) are two zinc metalloproteinases involved in regulating the metabolism of various peptides, and are particularly involved in blood pressure and fluid homeostasis. Controller (Fourni6-Zalusld et al. (1994 J. Med. Chem. 37: 1070-1083). ACE is a zinc-containing carboxydipeptide that converts the inactive precursor angiotensin I to angiotensin II, a peptide that promotes vasoconstriction and sodium retention, and therefore increases blood pressure. Compounds with ACE inhibitory activity are useful in the treatment of hypertension, heart failure, and post-infarction. NEP (also known as, brain "Enkephalinase") is a zinc-containing endopeptidase that is found at high concentrations in the brush border region of the kidney. NEP causes atdal natriuetic factor (ANF) loss ANF is a hormone secreted by the heart, which increases vasodilation, and increases urine and natriuresis at the kidney level. It has a compound that inhibits the activity of neutral endopeptidase (nep) enzymes. , 疋 used for blood Dilators and NEp are responsible for the reduction of bmdykinin of the vasomotor peptides acting on the endothelium and epithelium. Therefore, when ACE and NEp exert their effects on the cardiovascular system through different mechanisms of action Compounds with mixed ACE-NEP inhibitory activity are usually used alone or in combination to treat hypertension, f

(210 x 297 mm) 200407310 (A) A7 5. Description of the invention (2) Dirty failure, congestive heart failure, and ischemic heart disease. World patent WO 97/24342, which is hereby incorporated by reference, discloses certain sulfhydryl phenyl phenylalanine derivatives of formula (A), which have ACE-NEP inhibitory activity, and are used for treatment Cardiovascular disease mountain 17 π · South blood pressure 5 and congestive heart failure. J1 Η R—C——C—C〇NH—C—C00R2 h2 h I * C—R3 H2 Among them: 10 Consumers' cooperation of Intellectual Property Bureau of the Ministry of Economic Affairs, printed by society 15 20 R is a mercapto group, or it can be used in organisms R4C0S group which can be converted into thiol group in the group; R! Is a straight-chain or branched Q-C4 alkyl group, aryl group or aralkyl group, having from 1 to 6 carbons in the alkyl portion Atoms in which the aryl group is phenyl or a 5- or 6-membered aromatic heterocyclic ring with one or two heteroatoms, the heteroatoms are selected from the group consisting of nitrogen, oxygen, and sulfur, optionally with one or two identical or Different substituents, selected from halogen atoms, hydroxyl groups, alkoxy groups, alkyl groups, alkylthio groups, alkylsulfo groups, or alkoxycarbonyl groups, having from 6 to 6 carbon atoms in the alkyl portion; containing one CrC3 grafting group or carboxyl group, nitro group, amine or amine carbonyl group, fluorenylamine group, or a group of one or more gas atoms The amine group has a group of 1 to 6 carbon atoms in the moiety. & is a hydrogen atom, straight chain or branched Cl ·. Alkyl & is a phenyl group substituted with a 5- or 6-membered aromatic heterocyclic ring. The heterozygous group is selected from this paper. It is applicable to Chinese National Standard (CNS) A4 specifications (^ (297 200407310 A7) B7 V. Description of the invention (3) The group consisting of nitrogen, oxygen and sulfur, the phenyl or heterocyclic group may be optionally substituted with one or two same or different substituents, as shown. R4 is a straight chain Or a branched CrC4 alkyl group or phenyl group; the carbon atom marked with an asterisk is a stereogenic center; 5 and its pharmaceutically acceptable salts. Surprisingly, it has been found that: Compounds, which are generally disclosed in World Patent WO 97/24342, and have specific substituted forms, show improved properties over those compounds disclosed in World Patent WO 97/24342. 10 SUMMARY OF THE INVENTION Accordingly, the present invention Provide a compound of formula (I): VIII.R2

Ϊ1 Η (I) Among them: Printed by the Consumers ’Cooperative of the Intellectual Property Bureau of the Ministry of Acne 15 R stands for Ci-C6 alkyl; R2 stands for azole or pyrimidinyl; or a pharmaceutically acceptable derivative thereof. Another aspect of the present invention is .... _ Pharmaceutical composition containing 20 compounds from the compound of the present invention and a pharmaceutically acceptable carrier and / or excipient 200407310 A7 B7 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Description of the invention (4)-A compound of the invention for use in therapy. -Use of a compound of the present invention for the manufacture of a medicament for the treatment of a patient suffering from a disease which may be ameliorated with ACE and / or NEP inhibitors. -A method of treating a patient suffering from a disease that may be ameliorated by an ACE and / or NEP inhibitor, comprising administering a compound of the invention in a therapeutically effective amount. DETAILED DESCRIPTION OF THE INVENTION The compound of formula (I) contains a palm (asymmetric) center (indicated by *). Individual stereoisomers (enantiomers and diastereomers) and mixtures of these are within the scope of this invention. As used herein, the term "alkyl" means straight and branched chain saturated hydrocarbon groups. Examples of the alkyl group include methyl, ethyl, propyl, and butyl groups. Preferably, R1 represents a Q-4 alkyl group, more preferably, Ri is a C3-4 alkyl group, and most preferably isopropyl. 15 R2 represents pyrazole or pyrimidine. Pyrimidine is C-linked to a benzene ring. Pyrazoles can be N-linked or C-linked to a benzene ring. Preferably, R2 represents. Optimally, R2 stands for N-linked Hou 17 sit. It is to be understood that the invention encompasses all combinations of suitable, convenient and preferred groups described herein. 20 The compounds of the present invention show improved inhibitory activity on human plasma ACE, together with good inhibitory activity on NEP, and thus achieve better efficacy in humans. As used herein, the term "mixed ACE-NEP inhibitor" means a compound having two inhibitory activities, 'ACE and NEP. The term "double" has become more general -6- This paper size applies to the Chinese National Standard (CNS) A4 specification (2 [() x297 mm)

20〇4〇731〇 A7 B7

Ground is used in the literature. For the purposes of this patent application, the terms mixed and dual are applied equivalently. As used herein, the term "pharmaceutically acceptable" means a compound suitable for medical use. ) As used herein, the term "pharmaceutically acceptable derivative" means any pharmaceutically acceptable salt, solvate, or prodrug of a compound of formula (i), for example: 'It can (directly Or indirectly) provide a compound of formula (I), or an active metabolite or residue thereof. Such derivatives are recognized by those skilled in the art without undue experimental theory. However, reference is made to Burger's Medical Chemistry and Drug Discovery, 5th edition, Volume 1: Principles and Practice, for the extent to which this derivative is taught here and here reference. Preferred medically acceptable derivatives are salts, solvates and esters. Particularly preferred pharmaceutical acceptable derivatives are salts and solvates. 15 Examples of medically acceptable salts are salts of metal or earth metals and salts with pharmaceutically acceptable organic bases. See Berge et al., J. Pharm. Sci. 1977, 66, 1-19, which is incorporated herein by reference. Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economics. Those skilled in organic chemistry will understand that many organic compounds can form complexes with solvents, which react in or form precipitates or crystals from the solvents. 20 These complexes are known as solvates. For example, the complex with water is known as a hydrate. The solvate of the compound of formula (I) is within the scope of the present invention. Salts and solvates of the compounds of formula (I) suitable for use in medicines are those in which counterions or related solvents are pharmaceutically acceptable. However, they are non-medical. It is within the scope of this paper to apply the Chinese National Standard (CNS) A4 specification (210x 297 meals) 200407310 A7 _______ B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Description of the invention (6) The scope of the invention, for example: It is used as an intermediate for preparing other compounds of formula (I) and its pharmaceutically acceptable salts and solvates. As used herein, the term "prodrug" means a compound that is transformed in the body, for example, hydrolyzed in blood to become its active form with medical efficacy. 5 Medically acceptable predrugs are described in T. Higuchi and V. Stella as Prodrugs as Novel Delivery Systems' Symposium Series Volume 14 and in Edward B. Bioreversible Carriers m Drug Design edited by Roche, American Pharmaceutical Association 10 and Pregamon Press, 1987, both of which are incorporated herein by reference. Esters can be active in their own right and / or hydrolysable in the human body under living conditions. Suitable pharmaceutically acceptable ester groups that are hydrolysable in vivo include those which are easily cleaved in the human body leaving behind the mother acid or its salt. 15 Preferred compounds according to the present invention include and may be selected from the following: N-[(2S) -2-[(mercaptomethyl) -3-methylbutylfluorenyl] -4- (1fluorene-pyrazolyl) phenyl Alanine; N-[(2S) -2-[(mercaptomethyl) -3-amidinopentyl] -4-pyrimidin-5-yl-L-phenylalanine; 20 N-[( 2S) -2-[(Mercaptomethyl) -4-fluorenylbutanyl] -4- (1fluorene-pyrazol-5-yl) -L-phenylalanine; N-[(2S) -2- [ (Fluorenylmethyl) -3-fluorenylbutylfluorenyl] -4- (1fluorenyl-pyrazol-5-yl) phenylalanine; ^ N-[(2S) -2-[(fluorenylmethyl)- 3-methylbutylfluorenyl] -4- (1Η-pyrazol-4-yl) -L-benzene This paper is sized for China National Standard (CNS) A4 (210 X 297 mm)

200407310 five

Alanine; and its pharmaceutically acceptable derivatives.

Particularly preferred compounds according to the present invention include and may be selected from the following N-[(2S) -2-[(mercaptomethyl) -3-methylbutylfluorenyl> 4- (1H.pyrazoldiphenyl) ^ benzene ] Alanine; '& [(2S) -2-[(Mercaptomethyl) -4-methylpentamyl]]-4- (1Η, pyrazolyl) -alanyl alanine; 卞Team [(2S) -2-[(Mercaptomethyl> 3-methylbutyridylbiazole, 5-yl) a-benzylalanine; Team 10 ((2 外 2 · [(薇基 methyl 甲 甲Butylfluorenyl] Bipyl_4_yl) Cardiobenzyl Alanine; and its pharmaceutically acceptable derivatives. Inhibitors, and therefore diseases, such as: cardiovascular compounds of the present invention are mixed ACE / NEP in Treatment with ACE and / or NEP inhibitors can improve 15 diseases and kidney diseases. The compounds of the invention show superior properties, and the whole product of πw ~ HuΊM is more effective. Large selectivity, less side effects, 'longer manufacturing time, more biologically effective, or other more desirable properties through a better route. 20 Therefore, the present invention provides a compound of formula (I) or a pharmaceutically acceptable compound thereof. Derivatives, especially for humans The invention also provides another aspect of the present invention. The compound of formula (I) or a medicament thereof. The use of the derivative can be used for the treatment of a disease that may be improved by an ACE and / or NEP inhibitor. 200407310 A7 Consumer Co-operation, Intellectual Property Bureau, Ministry of Economic Affairs, Printed by the Society V. Invention Description / In another or additional aspect, a method for treating a human-feeding animal is provided, which comprises administering an effective amount of a compound of formula (I) or a compound thereof Derivatives can be used, especially for the treatment of diseases that may be improved with ACE and / or NEP inhibitors.) It should be understood that the referenced treatment is intended to include the prevention and alleviation of established symptoms. The compound of formula (I) can be applied as a raw chemical, but the active ingredient is preferably presented as a pharmaceutical formulation. Therefore, the present invention further provides a medicinal formulation composition comprising at least one compound of the formula (I) or a pharmaceutically acceptable derivative thereof, and a related west-acceptable carrier and / or excipient. The carrier and / or excipient must be cross-linkable "'meaning to be compatible with the other ingredients of the formulation and not harmful to the recipient thereof. In another aspect, the present invention provides a pharmaceutical composition Containing at least one compound of formula (I) or a pharmaceutically acceptable derivative thereof as an active raw material, and a pharmaceutically acceptable carrier and / or excipient associated therewith, for the treatment of sub- and 4-inches, and for the treatment of patients who may suffer from ACE and And / or NEp inhibitors to change the disease in human or animal patients. What is provided by the present invention is a method for preparing a pharmaceutical composition, each of which is a compound of formula (I) or a pharmaceutically acceptable derivative thereof, It is mixed with medically acceptable carriers and / or excipients. Compounds of formula (I) can be formulated for oral, oral, parenteral, ophthalmic and nasal use), storage (dePQt), or rectal administration. It is administered by inhalation or insufflation (oral or nasal). For oral administration, the pharmaceutical composition can be, for example, tablets: Standard (CNS) A4 Specification -10- 200407310

10 15 Employees from the Intellectual Property Bureau of the Ministry of Economic Affairs congratulate the cooperation and exemption. 20 制备 Prepare the supplementary with a pharmaceutically acceptable carrier such as: adhesives (e.g., pre-gelatinized corn starch, polyvinyl tetrazolide) Or methyl ethyl cellulose); fillers (for example: lactose, microcrystalline cellulose or hydrogen phosphate); lubricants (for example: stearin _ silicon dioxide); disintegrants (for example: potato powder or potato; Powder sodium glycolate): or shore wetting agent such as: sodium lauryl sulfate). The tablets can be coated in this manner in a well-known manner. Liquid preparations for oral administration may take the form of, for example, a solution, a sugar pack or a shoe float, or they may be presented as dry products formulated with water or other suitable vehicles before use. Such liquid preparations can be prepared in a customary manner and with pharmaceutically acceptable additives such as: suspending agents (for example: sorbitan sugar water 'cellulose derivatives or hydrogenated edible fats), emulsifiers (Eg, phospholipids or gum arabic); non-aqueous vehicles (eg, almond oil, oily esters, ethyl alcohol, or fractionated vegetable oils); and preservatives (eg, methyl or propyl p-hydroxyl Benzoate or sorbic acid). The formulation may also contain suitable buffer salts, flavors, colorants and sweeteners. Formulations for oral administration can be suitably formulated to produce controlled release of the active compound. For oral administration, the composition may take the form of tablets or lozenges formulated in a conventional manner. The compounds according to the invention can be formulated or administered parenterally ', e.g. by injection or continuous injection. Formulations for injection can be presented in single-dose form, e.g. in ampoules or multi-dose containers with preservative M added. The illegal composition can be in the form of a suspension, solution or emulsion in an oily or aqueous medium, and contains preparation reagents, such as: hanging bound paper ruler " -11- 200407310 A7 B7 ---- --- '^' -------- 5. Description of the invention (10) Floating, stabilizing and / or dispersing agent. Alternatively, the active ingredient may be in the form of a powder or made up of a suitable vehicle before use, such as sterilized pyrogen-free water. The compounds according to the invention can be formulated for topical application by inhalation or inhalation 5. Examples of the types of preparations for topical application include mouth sprays and aerosols for use in an inhaler or inhaler. Topical powders are formed with the aid of any suitable powder base, such as: lactose, talc or starch. The spray composition can be formulated as an aqueous solution or suspension or an aerosol delivered by a pressure pack, such as a suction device for measuring doses and the use of appropriate propellants. The compounds according to the invention can also be formulated into rectal compositions, such as: pick-ups or retention enemas, for example, containing conventional suppository bases, such as cocoa buffers or other glycerides. Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs In addition to the aforementioned formulations, the compound can also be formulated into storage formulations. This long acting formulation can be implanted (for example: subcutaneously, transdermally or intramuscularly) for application or intramuscular injection. Thus, for example, the compounds according to the present invention can be formulated with suitable polymeric or hydrophobic substances (e.g., emulsions in acceptable oils), ionomer resins, or low-soluble derivatives, such as low-soluble salts. The daily dose of a compound of formula VII or a pharmaceutically acceptable derivative depends on more than 20 factors, such as the severity of the disease, the individual response of the patient, or the type of formulation, but it is usually included per kilogram of body weight Between 0.1 mg and 10 mg, divided into single or more daily doses. Compounds of formula (I) may also be used in combination with other therapeutic agents. Therefore, in another aspect, the present invention provides a compound containing the formula VII or a pharmaceutically acceptable -12-standard scale ^ standard "... Regulations ^ · ^ ^ ^; 200407310 V. Description of the invention (" A combination of a derivative thereof and another therapeutic agent. When the compound of formula (I) or a pharmaceutically acceptable derivative thereof is used in the same disease condition, the brother of a sadness-therapeutic active agent, the amount of Jiuhuaba #, '6 may be different. When the compound is used alone. The compound of the present invention is used in combination with ACE and / or NEP inhibitors and the like. However, the above-mentioned combination can be conveniently presented in the form of a pharmaceutical composition, and includes The combination of the above definitions and pharmaceutically acceptable carriers or excipients contains another aspect of the present invention. The second component of this type: the respective components of the second component can be combined with the second component: and 5 10 Application, by any convenient route. / ,,, and when continuous application, mixed ACE_NEP # 卩 · silk two agents can be applied first. When applied simultaneously, the combination can be applied with the same or two components. Shangshi 15 When combined in the same formulation It must be understood that the two compounds must be stable and compatible with each other and the other components in the formulation. When formulated separately, 'it can be provided by any convenient formulation, conveniently for such compounds in this formula. This is a known method. Printed by the Consumers' Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 20 fields of the compound of formula (I) or its medically acceptable derivative are used as the second treatment for the same disease state as with live confrontation When the reagents are combined, the compound may be different from the compound when used alone. The appropriate dosage can be easily understood by those who know it. It is understood that the compound of the present invention that is needed for the treatment is required. The nature of the disease to be treated and the age and state of the patient change, and are ultimately the consideration of the participating physician or veterinarian. Compounds of formula (I) and their pharmaceutically acceptable derivatives can be described later by -13- + Gongchu) 200407310 A7 B7 V. Description of the invention (12) Preparation of the process, which constitutes another aspect of the present invention. In the following description, this group is a compound of formula (I) as defined above, unless In other words, when the compound of the present invention is administered in the above-mentioned dose range, a non-toxic effect is indicated / expected. 5 According to another aspect of the present invention, a process (A) for preparing a compound of the formula (I) is provided. The process includes reacting a compound of formula (II) with a compound of formula (III) for deprotection:

I

L H2r ^ c〇2pi / ττλ

10 (III) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs, where R1 and R2 have the above meanings, and P1 represents an oxygen protecting group, for example, fluorenyl. The condensation reaction is performed using conventional techniques of peptide chemistry. Deprotection reactions are performed using conventional techniques. Suitably, the reaction can be carried out in the presence of a coupling reagent, for example: 1- (3- (bis (15 fluorenylamino) propyl-3-ethylcarbonyldiimidoamine hydrochloride), In the presence of hydroxybenzyltriamidine), in a suitable solvent, for example: DMF (N, N-dimethylamidamine), MeCN, DCM, preferably DMF, suitably at room temperature. In addition, the reaction can be borrowed The method is to activate thionyl chloride to activate the compound of formula (III), and then continue to invert the activated compound of formula (III) and compound of formula (II) in a suitable solvent. CNS) M specification (210 x 297 mm) 200407310 A7 B7 V. Description of the invention (Π) The solvent is, for example, DCM, ethyl acetate, preferably ethyl acetate, and the test is, for example: K: 2C03 In the presence of. The reaction is continued to deprotect under standard conditions, such as ... When P1 represents a Cw alkyl group, the removal of the protective group can be NaOH in a solvent such as: THF (tetrahydrocraw), or suspended in Me OH. Di-5 sulfide removal can be achieved by tributylphosphine treatment. Then compound i of formula (I) is treated with an acid such as HC1 and deposited. Formula (III) The compounds are known or can be prepared according to conventional methods described, for example, in British Patent No. 1,576,161 and Fournie-Zalusky et al. (1994) J. Med. Cliem. 37: 1070-1083 'each of which is incorporated herein by reference Reference 10 teaches compounds of formula (III) and the extent to which they are prepared. In addition, compounds of formula (III) can be prepared from compounds of formula (XI): 〇〇

Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs Reacts with acids such as HC1, suitably in the presence of solvents such as toluene or DCM. 15 A compound of formula (XI) can be prepared from a compound of formula (XII): 〇 〇

Reacts with ephedrine in the presence of a solvent such as isopropyl acetate. Compounds of formula (XII) can be prepared from compounds of formula (XIII): -15- This paper size applies to China National Standard (CNS) A4 (2 丨 Ox 297 mm) 200407310 A7 B7 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs V. Explanation (14

(XIII) reacts with thioacetic acid in the presence of a suitable catalyst such as Cs2C03, K2CO3, Na2C03, preferably Cs2C03, and in a solvent such as MIBK Exist. Compounds of formula (XIII) can be prepared from compounds of formula (XIV):

10 ACT Ya, 0A (XIV) where A is Ci_6 alkyl, by hydrolysis of the ester group under standard conditions such as using NaOH, continued with the presence of secondary amines such as diamidoamine The reaction with formic acid (Manmcli reaction), followed by neutralization with acid such as HC1. According to process (B), compounds of formula (II) where R2 is N-linked pyrazole can be prepared from compounds of formula (IV):

? H

ΌΗ 15 (IV) where P1 is an oxygen protecting group, for example: fluorenyl, B represents boron, and P2 is an amine protecting group, such as: Boc, in copper acetate, pyridine, and TEMPO (tetramethyltetrahydro ° ratio ° Each ketone oxide) is treated with pyrazole in an organic solvent such as DCM (dichloromethane, j: end); continued with deprotection of amine groups under standard conditions. -16- The size of this paper applies to Chinese National Standard (CNS) A4 (2 丨 0 X 297 mm) 200407310 A7 B7 V. Description of the invention (15) The compound of formula (IV) is known in the art, see for example: M. E. Jung, TI Lazarova, J. Org. Cliem. 1999, 64, 2976, which is incorporated herein by reference, teaches compounds of formula (IV) and the extent to which they are prepared. According to process (C), the compound of formula (II) wherein R2 is a C-5 linked pyrazole 5 can be prepared from a compound of formula (V):

L 1 (V) where P1 is an oxygen protecting group, for example: methyl, SEM is 2- (trimethylsilyl) ethoxy] methyl, and P2 is an amine protecting group, such as: Boc, in Such as reflux in a solvent of ethanol in the presence of an acid such as HC1. 10 Compounds of formula (V) can be prepared from compounds of formula (IV) by reacting with compounds of formula (VI):

SEM (VI) Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs, where SEM is 2- (trimethylsilyl) ethoxy] fluorenyl, in the presence of a base such as potassium carbonate, for example: DME In a solvent such as: PdCh metal 15 catalyst, under elevated temperature. The reaction is preferably carried out at 30-100 ° C, more preferably at about 70 ° C. Compounds of formula (V) can be prepared from compounds of formula (VII): -17- This paper size is applicable @National Standard (CNS) A4 specification (210 X 297 mm) 200407310 A7 B7 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs

V. Description of Invention (16)

(VII) In the presence of tetrahydrofuran and n-butyllithium, at a temperature below room temperature, preferably _78-0 ° C, and react with iodine. Compounds of formula (VII) are known in the art, see for example: N. Fugma, W. 5 Holzer, M. Wasicky; Heterocycles, 1992, 34, 303, which is incorporated herein by reference to teach compounds of formula (VII) And the extent of their preparation. According to process (D), compounds of formula (II) in which R2 is pyrimidine can be prepared from compounds of formula (VIII):

L (VIII) 10 where P1 is an oxygen protecting group, such as methyl, and p2 is an amine protecting group, such as Boc, in a solvent such as dioxolane, nitrogen, and room temperature, and HC1 reaction. Compounds of formula (VIII) can be prepared from compounds of formula (IV) and react with 5-bromopyridine in the presence of a base such as potassium carbonate and a metal catalyst such as PdCl2. The reaction is carried out at elevated temperature, preferably at 30-100 ° C, and more preferably at about 50 ° C. -18- This paper size applies to China National Standard (CNS) A4 (210 x 297 mm)

200407310 A7 B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs (IX) 'and P' is an amine-protecting group V. Description of the invention (17) According to process (E), where R2 is a 4-C4-linked pyrazole formula (II) Compounds can be prepared from compounds of formula (IX): / Ph,

I

U p2Nir ^ c02Pl where P1 is an oxygen protecting group, for example: methyl 5 group, such as: Boc, in a solvent such as dioxolane, nitrogen, at room temperature, and react with HC1. A compound of formula (IX) can be prepared by reacting a compound of formula (IV) with a compound of formula (X):

10 In the presence of a base such as potassium carbonate, in a solvent such as DME and in a metal catalyst such as PdCl2, under elevated temperature, and under nitrogen. The reaction is preferably carried out at 30-100 ° C, more preferably at about 70 ° C. Compounds of formula (X) are known in the art, see for example: J. Elguero, C Jaramillo, C. Pardo; Synthesis, 1997, 563), which is incorporated herein by reference to the teachings of compounds of formula (X) and 15 The extent of its preparation. According to the process (F), the compound of formula (II) can be prepared from the compound of formula (XV): -19- This paper size applies to the Chinese National Standard (CNS) A4 specification (210 x 297 mm)

200407310 A7 B7 V. Description of the invention 18 R2 P2NH '

X το〇Η (χν) 10 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 15 where P2 is, for example, an amine-protecting group of CO, in the presence of HC1, for example, under standard conditions of MeOH treatment Deprotect the amine group and continue to protect the carboxylic acid group in the presence of HC1, for example, under standard conditions of reaction with 1 ^ 60. Compounds of formula (XV) are prepared by compounding formula (XVI): I2 Λ Ρ2ΝΗ ⑴ Η 2Η (χνΐ) where P2 is, for example, an amino protecting group such as COH or hydrogen, and X is, for example: a halogen leaving group Group, preferably iodine; reaction with a compound of formula (XVII): R2-hydrazone (XVII) where R2 is pyrazole or pyrimidine, preferably pyrazole, in a transition metal catalyst such as Cul and for example: Cs2C03 In the presence of K2C03 base, Cs2C03 is preferred, and in a solvent of NMP (n-methyltetrahydropyridinone), 1,4-dioxolane, and DMF, NMP is preferred. Compounds of formula (XVII) are known in the art and are commercially available Compounds of formula (XVI) can be prepared from compounds of formula (XVIII): -20-

This paper size applies to China National Standard (CNS) A4 (210 x 297 mm) 200407310 A7 B7

X

L H2N C02pl (XVIII) where P1 is, for example, a carboxylic acid protecting group of a fluorenyl group, and p2 is, for example, an amine protecting group of COH or hydrogen, and X is, for example, a leaving group of a dentin, preferably Iodine; by deprotection of the oxygen group, under standard conditions such as reaction with NaOH, in a suitable solvent such as MeOH. Compounds of formula (XVIII) can be prepared from compounds of formula (XIX):

(XIX) 10 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 15 By reacting with X in the presence of peracetic acid and acids such as H2SCU 'and continuing to deprotect carboxylic acid groups under standard conditions, In the presence of, for example, MeOH, in the presence of an activator such as SOC12 and a solvent such as toluene, it can optionally be continued to protect the amine group under standard conditions, and react with, for example, HCOOH, in such as : In the presence of an activating group of acetic anhydride. Compounds of formula (XIX) are known in the art and are commercially available. In addition, compounds of formula (XVI) can be prepared from compounds of formula (XIX) by reacting with, for example: X of 12 in In the presence of acetic acid and acids such as H2SO4, and continued deprotection with amine groups, it is reacted with, for example, HCOOH under standard conditions, in the presence of activating groups such as acetic anhydride. -21- This paper size applies the national standard (CNS) A4 specification (210 X 297 mm) 200407310 A7 B7 V. Description of the invention (2 ()) The compound of formula (II) is a novel compound and therefore forms the Another point. Those skilled in the art will understand that in the clothing of the compound of formula (I) or its bath sword product, it needs and / or wants to protect one or more sensitive groups in the molecule ' to avoid unwanted side reactions. The protecting group used for the preparation of the compound of the formula (I) can be used in a conventional manner. See, for example, Protective Groups in Organic Chemistry (Protective Groups m Organic Chemistry, JFW McQmie, Edited by the Consumer Cooperatives of the Intellectual Property Bureau, Ministry of Economic Affairs, Plenum Press, London (1973) or Protective Groups in Organic Synthesis) m Organic Synthesis), Theodora Green, 10 John Wiley and Sons, New York (1981). Examples of suitable amine-protecting groups include fluorenyl-type protecting groups (for example: formamyl, trifluoroacetamidine, acetamidine Group), protecting groups of aromatic amino phosphonates (for example: benzyloxycarbonyl (Cbz) and substituted Cbz), protecting groups of aliphatic urethanes (for example: 9-fluorenyl ( fluorenyl), fluorenyloxycarbonyl (Fmoc), tertiary-butoxycarbonyl 15 (Boc), isopropyloxycarbonyl, cyclohexyloxycarbonyl) and alkyl protecting groups (for example: benzyl , Trityl, chlorotribenzyl). Examples of suitable oxygen protecting groups can include, for example: alkylsilyl groups, such as trimethylsilyl or tert-butylfluorenylsilyl; Ethers, such as: tetrahydroxypyranyl or tert-butyl; or esters, such as Acetate. 20 The following examples illustrate the viewpoints of the present invention, but must not be construed as limiting the scope of the present invention in any way. Example 1 The combination of intermediates in Example 1-22- This paper applies Chinese National Standard (CNS) A4 Specifications (210 ^ 297) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 200407310 A7 B7 V. Description of Invention (21

BocNH ^ ^ CO-Me 丄

BucNH ^^ COiMe HC1 .Fi2N ^ vC〇2Me 5 Intermediate 2: N- (Third-butyridylcarbonylpyrazol-1-yl) -L-benzylalanine methyl ester boric acid * (1) (1.〇. G, 3.09 mmoles), pyrazole (0.2 g, 3.09 mmoles), copper acetate (0.82 g, 4.64 mmoles), pyridine (0.5 ml, 6.18 mmoles), TEMPO (0.482 g , 3.09 mmol), and 4A molecular sieve 10 (1/8 ", dry), stirred at room temperature in dichloromethane (100 ml) for two weeks. The mixture was then filtered through hyflo and in vacuo The solvent was evaporated. Purification by silica gel chromatography (dichloromethane / cyclohexane 1: 1 to dichloromethane / ethyl acetate 1: 1) gave the title compound (0.675 mg) as a yellow solid. 15 LCMSRT3. 16 minutes MH + 346 * ME Jung, TI Lazarova, J. Org. Chem. 1999, 64, 2976 Intermediate 3: 4- (lH-. Biazol-1-yl VL-benzyl alanine phosphonium ester 20 at 4 moles of HC1 in dioxin (10 ml) was added to a solution of carbamate (2) (675 mg, 1.96 mmol) in dioxin (10 ml). After 4 hours at room temperature, the solvent was removed. Evaporate, and the residue is co-evaporated with diethyl ether (2x30 ml) to produce the title compound. LCMS RT 1.97 minutes MH + 246 (for free base) -23- This paper applies the Chinese National Standard (CNS) A4 (2U) x 297 mm)

200407310 A7 B7 V. Description of the invention (22) Synthesis of intermediate of Example 2

Intermediate 4: N- (fluorenedi-butoxydicarbonyl) -4-methylidene-5-yl-L -.- phenylpropanylsulfonium methylsulfonic acid boric acid * (1) (3.0 g ), 5-bromopyridine (1.77 g), 10 PdCl2 [dppf] (0.39 g) and potassium carbonate (5.1 g) were mixed in degassed DME (45 ml). The mixture was heated to 50 ° C for 3 hours and then cooled to room temperature. The mixture was diluted with saturated aqueous ammonium chloride (25 mL) and concentrated in vacuo. The residue was distributed in water and ethyl acetate. The organic portion was dried over sodium sulfate and the solvent was evaporated in vacuo. Silica gel chromatography (ethyl acetate 15 / cyclohexane 1:10) yielded the title compound (1.13 g). LCMS RT 2.92 minutes MH + 358 E. Jung, T. I. Lazarova, J. Org. Chem. 1999, 64, 2976 Intermediate 5: 20 4- ° dense printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs. Amine-5-yl-1 ^ -phenylalanine 1 芰 曱 §carbamate (4) (1.12 g) was dissolved in dichloromethane (20 ml), and 10 ml) of 4 mol HC1. The mixture was stirred at room temperature for 18 hours. The solvent was evaporated in vacuum, and the remaining residue was co-evaporated three times with dichloromethane (20 ml) to produce the title compound -24- This paper size applies to China National Standard (CNS) A4 (2U) X 297 mm 200407310 Α7 ------- B7 V. Description of the invention (23). LCMSRT 1.89 minutes MH + 258

〇ί SEM 6 Intermediate 7: 1〇 ^ blanket-liLULgj-based silicon-based) Ethyl-N-methylpyrazole A pyrazole Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Co-operative Society Printed the experience of ItJ in tetrahydrofuran (20 ml) Protect Π than α sitting # (6) (10 g, 5.04 mol) under nitrogen ... 78. 〇, n-butyl (H.29 mmol) in hexane was added, and the mixture was searched for one hour. E 0.27 g, 5.04 moles) was added as a tetrahydrocracking solution (10 ml). And the mixed mixture was stirred at room temperature for one hour. Water (50 ml) was added to the mixture, which was extracted with ether (2 x 50 ml). The organics were dried over sodium sulfate and the solvent was evaporated in vacuo to give an oil. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether 40-60 M 9) to give the title compound (0.88 g) as an oil. > 25- This paper size applies Lao National Standard (CNS) A4 specification (210 X 297 issued) 200407310 A7 B7 V. Description of the invention (24 LCMS RT 3.64 minutes (blue), MH + 325 #N. Fugina, W. Holzer, M. Wasicky; Heterocycles, 1992, 34, 303. Intermediate 8: N- (Third-butoxy, a carbonyltrisylsilyl group), Ethyl gas, a methyl chloride, 1H-azole-5- -L · · phenylalanine methyl ester 10 15 Consumption cooperation with employees of the Intellectual Property Bureau of the Ministry of Economic Affairs, printed 20 pairs of deaconazole intermediates (7) in DME (5 ml) (0.5 g, 1.54 mmol) and boric acid * (1) (0.415 g, 1.28 mmol), a solution of carbonic acid (0.89 g, 6.43 mmol), and PdCl2 [dppf] (0.53 mg, 0.06 mmol) were added. The mixture was stirred at 70 ° C for 16 hours. After cooling to room temperature, the solvent was stripped in vacuo, and the residue was distributed in ethyl acetate and water. The organic portion was dried over sodium sulfate, and Removal of the solvent in vacuo gave a crude product. Silica gel chromatography (ethyl acetate / petroleum scale 40-60 L4) yielded the title compound as an oil (380 mg). LCMS RT 3.89 minutes (Blue), MH + 476 * M · E · Jung, T. I. Lazarova, J. Org. Chem. 1999, 64, 2976 Intermediate 9: Sialyl-5_ylbenzylalanine methylparaben The amino acid derivative (8) (0.38 g, 0.8 mmol) was added with ethanol (1 liter) and 2 equivalents of HC1 (12 ml). The reaction was stirred under reflux at $ 30. The mixture was then cooled, and ethanol was removed in vacuo, neutralized with potassium carbonate (saturated aqueous solution), and extracted with aerosol. Organic; Yu-26- This paper size is in accordance with Chinese National Standard (CNS) A4 (210 X 297 Public Love 200407310 A7 B7 77) Instructions 25 Dry on sodium sulfate and remove the solvent in vacuum to produce a gum. Purified by ion exchange chromatography (SCX-2) to elute with 10% ammonia in methanol to give the title compound (0.075 g).

Synthesis of LCMS RT bodies

CPh3 ο

5 It Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 2 Intermediate 11: Base-1-Sanbenjia-1H- 咄 4 4- / Small π ratio ί (ίο 克 '6_8mmol) and three Ethylamine (0.90 liters, 6.5 mmol) was stirred under nitrogen, DMF, ye. Benzoyl chloride (1.81 g, 6.5 mol) was added, and the mixture was stirred at room temperature: for two days, the mixture was then diluted with aerobic (10 mL), and then water: strip. The organic portion was dried over sodium sulfate and the solvent was evaporated in vacuo to give a crude product. The resulting solid was washed with diisopropyl ether to give the title compound (1.55 g). LCMS RT 4.09 minutes, [cph3] + 243 27-degree ruled paper

: National Standard (CNS) A4 specification (2 丨 () x 297 public love) 200407310 V. Description of the invention 26 Λ7 B7 ± ΜΜΛΙ ^ ～~ oxoyl) -4- (1-trimomethyl-ΐΗ -α than α-sityl 1-phenyl) to bromopyrazole (] ") in degassed DME (10 ml) ([0 g, 2.56 birch), boric acid (1) (0.7 G, 2.14 mmoles) and potassium carbonate (1.5 g, 10.7 mmoles) was added with PdCl2 [dppf] (0.088 g, 0.1 mmoles) and then the reaction mixture was under nitrogen It was heated for 7 (rc, 24 hours. Then the solvent was removed in vacuo and the residue was distributed between ethyl acetate and water. The organic portion was dried over sodium sulfate and the solvent was evaporated in vacuo. Crude product. Purified by silica gel chromatography (ethyl acetate / petroleum 40-60 1: 4) to give the title compound (0 74 g). LCMSRT 4.11 min, [CPh3] +243 15 Intermediate 13: group Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs of the Ministry of Economic Affairs, VL-Benzyl Alanine Methyl Ester (12) (0.74 g, 1.26 mmol) was dissolved in dioxolane (15 ml) 2 Mollon In HC1, and the reaction was stirred at room temperature under nitrogen for 24 hours. The solvent was evaporated in vacuo, and the mixture was analyzed by SCX-2 ion-exchange chromatography and stripped with 1: 9 ammonia: methanol to give the title compound. (0.2g). LCMS RT 1.87 minutes, MH + 246 Synthesis of intermediates of Examples 1-5-28- This paper size applies to China National Standard (CNS) A4 specification (210 X 297 mm) 200407310

ιο 15 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 20 Intermediate 15: ^… 4_ 卞 基. Fluorenyl) oxazole_ When titanium tetrachloride (1.0 mol concentration in dichloromethane, 105 liters, 0.10 * 5 mol) is added, 4m (3-methylbutanoic acid) _ U4) (26.1 g '0.1 mole) in dry dichloromethane (_ml), c, nitrogen lice, and the mixture containing the yellow precipitate was stirred for an additional 15 sides, and then diisopropyl was added dropwise. Ethylamine (19 ml, 0 mole) Maintain temperature below 5t. The resulting purple solution was stirred for 75 minutes, and then 1Λ 孓 trioxolane (99 g, ΐ, ", ear) 'was added in methane (60 ml) and stirred for another 10 minutes, and titanium tetrachloride was added. (10 ears / day in DCM, 105 strokes, 0.105 moles). The mixture was stirred for 30 hours, and then the reaction was stopped by adding saturated ammonium chloride (500 ml). Water (100 ml) and monochloromethane (100 ml) were added, and the water phase was taken again in ⑽ml: gas Yin Yuanyuan, the combined organic phases were dried on NaJO4, and re-emitted. Recrystallized from 30% dichloromethane / petroleum, yielding 18.7 g (64%): * 0 demolition -29- M's scale is applicable to China National Standard (CNS) A4 (210 x 297 mm) 200407310 A7 B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs. 5. Description of the invention (28) The title compound as a white solid. LCMS RT 2.94 minutes MH + 292 Similarly prepared (R)-(4-benzyl-3- (2-hydroxyamidino-4-methylpentyl) -oxa-5sialone. Synthesis example: DA Evans et al., Tetraliedron, 1988, 5525. Intermediate 16: (R) -2-hydroxymethyl-3-fluorenylbutanoic acid 10 vs. oxazolone (15) (23.4 g, 80.4 mmol) in THF (300 ml) ) 30% hydrogen peroxide (90 ml, 0.8 mol) was added. Then aqueous lithium hydroxide (1.5 mol concentration, 107 ml, 160 mmol) was added dropwise, and the mixture was allowed to warm to room temperature and stirred 3 Hours. Then potassium hydroxide (9 g, 160 mmol) was added and the mixture was heated at 60 ° C for 30 minutes, and then cooled to 15 ° C. A solution of sodium sulfite (100 g) in water (400 ml) It was carefully added and the mixture was concentrated to two-thirds the volume and distributed in water (200 ml) and chloroform (500 ml). The aqueous phase was extracted again with 2x100 ml aeroform and then at 5 Molar HC1 (200 ml) acidified. The product was extracted with ethyl acetate (1 x 400 ml, 2 x 250 ml), washed with brine (500 mmol 20 liters), dried over Na 2 SO 4 and And evaporated to give a solid (15 g). This was redissolved in THF (300 ml), potassium hydroxide (2 equivalents) was added, and the mixture was heated at reflux for 3 hours. The volume was reduced to 1/4, added Water (300 ml), and the mixture was washed with aerosol (3x300 ml), acidified and extracted in ethyl acetate (5x300 ml), and dried and solvent-shifted -30- This paper is in accordance with Chinese national standards ( CNS) A4 size (210 x 297 male f)

200407310 A7 B7 Printed by the Consumers' Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Disclosure of the Invention (29) Excluding 8.1 g (76%) of the title compound. (R) -2-hydroxymethyl-4-methylvaleric acid was prepared similarly. The analysis details are the same as the literature values. 5 Intermediate 17: (Ethylmethyl-3-methylbutanoic acid was added to triphenylphosphine (32 g, 22 mmol) in dry THF (300 ml) at 0 ° C, Diisopropyl azadicarboxylate (24 ml, 122 mmol), a white precipitate was produced, which was stirred for another 10 minutes. (R) -2-hydroxymethyl-3- in THF (100 ml 10 liters) Methyl butyric acid (8.09 g, 61 mmol) and thioacetic acid (13.1 ml, 183 mmol) were added dropwise to Mitsunobu reagent. The mixture was allowed to warm to room temperature and stirred for 2.5 hours. The mixture was concentrated to a third volume in vacuo and distributed in ethyl acetate (400 ml) and aqueous sodium bicarbonate (2 x 300 ml), and carefully acidified with a 5 mole concentration HC1. The product was DCM (3x300 mL) was extracted, the solution was dried over Na2SO4 and evaporated to give a clear oil (9.6 g, 83%). (S) -2-Ethylthiomethyl-4-methyl was prepared similarly Valeric acid 20 Intermediate 18: AM (2SV2-IT ethylsulfanylthio) fluorenyl1-3-fluorenylbutanyl 4-1H-pyrazol-1-yl) -L · -benzylpropane S for pair in dichloromethane (25 ml) (2S) -2- (Ethylthio) -3-methylbutanoic acid (4.37 g, 23 mmol) was added dropwise with thionyl chloride (2.2 ml, 30 paper standards applicable to Chinese national standards ( CNS) A4 size (2 [0 x 297 mm)

200407310 A7 B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs. 5. Description of the invention (30 mol), and the solution was stirred at room temperature under nitrogen for 20 hours, then concentrated in a vacuum, and Chlorotriane (10 ml) was azeotropic. Amine hydrochloride (6 g) was suspended in a stirred biphasic mixture of dichloromethane (80 ml) and water (50 ml), and potassium carbonate (丨 4.5 g, 5 105 mmol) was added at 0 ° C. ). When complete dissolution occurred, the gasified acid in dichloromethane (30 ml) was added and stirred at 0 ° C. For 30 minutes, and then at room temperature for 1.5 hours. Dichloromethane (200 ml) and water (100 ml) were added, and the aqueous phase was extracted again with dichloromethane (200 ml). The combined organic phases were washed with 2 molar HC1, brine (200 ml), dried over Na 2 SO 4 and the solvent was evaporated. This material was redissolved in dichloromethane, hexane was added and it was contracted in vacuo 'until crystallization started. Re-diluted with hexane and 6.6 g of the title compound was taken into account. LCMS RT = 3.12 minutes MH + 418 15 Intermediate 19 was prepared similarly: Ethylsulfanyl) fluorenyl1-3-fluorenylbutanyl} -4-pyrimidin-5-yl L-benzylalanine methyl ester LCMS RT = 2.88 minutes MH + 430 20 Intermediate 20: yV-(((2SV2-R ethylfluorenylthio) fluorenyl1-4-fluorenylpentylfluorenyl 4-4-fluorenyl-oxazol-5-yl) benzyl alanine Acetic acid ester LCMS RT = 3.15 minutes MH + 432 -32- Paper size applies to China National Standard (CNS) A4 (210 x 297 mm)

Chuan040731

Interstitial body 21:, A site on L-phenyl propionyl methyl chloride LCMS RT-2.76 minutes MH + 418 10 i Interstitial body 22: Glycoxyl, H ^ -benzyl alanine methyl ester LCMS RT = 2.93 minutes MH + 418 〇0 15

Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 20 23 24 AZA from methylthiomethyl ~ 3-methylbutyric acid (+) ephedrine salt 1 Intermediate 24: Diethyl isopropylmalonate ( 1 times by weight) and 2 moles of aqueous sodium vaporized solution (2.2 when 1 '5.45 times the volume), and added at 80 ° C and 5 ° C for an additional hour. At the completion of the reaction, the contents were acidified with concentrated HC1 (U3 times by weight) and 60% aqueous dimethylamine (0.41 times by weight) was added, followed by 37% by weight / volume of water-based formaldehyde (0.49 times by weight), and The reaction was then searched at 90 ° C for 16 hours. The solution is cooled to 20 ± 5 ° C, and concentrated HC1 is added (0.83 -JJ. The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210x 297 mm) 200407310 A7 B7 V. Description of the invention (32 10 15 Economy The Ministry of Intellectual Property Bureau employee consumer cooperative printed 20 weight) and MIBK (4 times the volume). The layers were separated, and the organic layer was washed with water (1 times the volume). The MIBK layer was then concentrated to about M times the volume under reduced pressure. Then cesium carbonate (0.039 times by weight) was added, and the mixture was heated at 40 ± 5 ° C. Thioacetic acid (0.36 times by weight) was added, and the mixture was stirred at 40 ± 5. (: Stirred for at least 12 hours. The reaction mixture was Cool to 20 ± 5 ° C and add potassium bicarbonate (2.25 times the volume). The layers are separated and the organic layer is washed with 20% potassium hydrogen hydroxide (0.9 times the volume). The combined aqueous layer is HC1 (1.18 times) Weight) adjusted to pH 1, and then the acidified layer was extracted with isopropyl acetate (2 × 2 times the volume). The combined organic layers were then washed with water (1 times the volume), and the mixture was concentrated to 2 times the volume by vacuum distillation. And then add isopropyl acetate (4 9 times the volume ). (+)-Ephedrine hydrochloride (0.86 times by weight) was suspended in isopropyl acetate (4.2 times by volume) and water (0.7 times by volume). 108 moles of hydrogen was added. Sodium oxide (0.58 times the weight) 'and stirred to produce a biphasic solution. The phases were separated and the organic phase was washed with water (0.35 times the volume), and the solvent was reduced to 3.5 times the volume by vacuum distillation. The added ephedrine solution (M Times the volume) into the racemic solution. The lower seeds are crystallized and added at 2 (TCn hours, then the remaining ephedrine is added within 2-3 hours. The crystals are cooled to 0 pits, and at least 2 hours k .; Yuzhong was filtered, and the filter cake was washed with isopropyl acetate (2 times volume) and dried in a vacuum oven at 50 ± 5 ° C. Expected yield: 33% of theory, 58% weight Weight. ', HPLC (2-minute method) RT 〇27 minutes 27 ％ area, ^ minutes 71.9% area. -34- This paper size applies Chinese National Standard (CNS) A4 specification (2 丨 × 297 mm 200407310 A7 B7 V. Description of the invention (3〇

15 Consumption cooperation by employees of the Intellectual Property Bureau of the Ministry of Economic Affairs, printed by the company 20 Glacial acetic acid (2.73 times weight, 2.62 times volume) Stir under 20 ° C, and add concentrated sulfuric acid (1.34 times weight, 0.73 times volume) while maintaining temperature Below 45 C. After this mixture was cooled to 20 ° C, a moth weight, 0.5 equivalent) was added, and L-phenylalanine (multiple weight) was added with stirring.混合物 The mixture was heated to 55 ± 5r, stirred by a pump, and 40% peracetic acid (about 0.75 times by weight) was added within 2-4 hours. Then check the library for completeness by M HPLC (<5% area of residual phenylalanine, generally ~ ^ 4). Polyoxide-deficient species are smelted with Merckoquant peroxide test strip, saturated sodium metabisulfite solution (minimum volume, _ &lt; ° added, known as 0.5, ι times volume) to convert any The remnants of E have become occupations. The iodized mixture was cooled to 35 ° C, vacuum was applied, and the batch 俾 ° moon bean product was reduced to about 4 times the volume by distillation. Add methanol (2 times the volume), and say ^% ^ 蒸 -35- The local scale is applicable to the Chinese National Standard (CNS) A4 specification (210x297 mm) 200407310 A7 B7 V. Description of the invention (34) Distillation until approval The secondary volume is reduced again to about 4 volumes. Add more methanol (2 volumes) and continue the distillation until the batch volume is reduced to about 4 volumes. Finally, methanol (2.5 times the volume) was added and the mixture was cooled to 20 ± 5 ° C. The mixture was used directly in the next step. 5 The suspension of the crude iodophenylalanine (about 1.8 times the weight and about 2 times the weight in methanol) from step la was heated to 50 ° C, and thionyl chloride (1.02 Weight, 0.63 times volume, 1.42 equivalents), and the temperature does not exceed 60 ° C at this rate. The mixture was then heated to 55 ± 5 ° C and maintained at 55 ° C for a minimum of 2 hours. The reaction was then checked for completion by 10 HPLC (&lt; 5% area of residual iodophenylalanine, typically 2%). The temperature of the mixture was adjusted to 40 ° C and distilled under vacuum until the volume was reduced to about 3 times the volume (maintain a minimum temperature of 40 ° C during this operation). The mixture was then diluted with toluene (1 volume) and maintained at a minimum temperature of 15 30 ° (:, continued with water (2.5 volumes). The mixture was cooled to 20 ° (:, and printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs) The force is 0.880 ammonia and (4 times the volume, 3.52 times the weight) and toluene (1 times the volume), while the temperature is kept below 35 ° C. Then the mixture is allowed to stand. The toluene layer is removed and the water layer is again toluene. (1 volume) extraction. The organic layers were combined and distilled under vacuum to produce a flowing oil. Toluene (0.5 fold 20 volume) was added and the organic solution was used in the next step without further treatment. Formic acid (1.02 Times the weight, 0.84 times the volume) and toluene (0.5 times the volume) at 12 ± 3 ° C, stir, and add acetic anhydride (0.86 times the weight, 0.8 times the volume) at 10-15 ° C, and the mixture is at Stir for an additional 60 minutes at 10-15 ° C. Then the toluene solution of the crude iodomethyl methyl ester from the previous step is at 10-15 -36- This paper size applies to China National Standard (CNS) A4 (210 X 297 mm_ Love) 200407310 A7 V. Invention Description c &quot; μ plus, And the resulting orange-brown mixture was stirred for at least 1 hour under 2 soil, and then the reaction was checked for completeness by HPLC (&lt; 3% of the area of residual stone methyl ester, generally &lt; 2%). To about 2 times the volume. The oil was diluted with methane (5 times the volume) and washed with water (4 times the volume), which was adjusted by adding 0.880 ammonia solution and then water (5 times the volume). Yang $. 15 15 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 20 Ran-chloromethane solution is diluted with toluene (4 times the volume) and concentrated under atmospheric pressure until it reaches 6-7 times the volume of the batch, generally At 65-7 ° C / valley solution. However, the cross-linking solution was cooled to 2q ° C (if seeding is required) in about 1 hour and then for 1 hour to 7 ° C. The crystallized product was at least 丨 hour Internal aging 'and transition under pressure. The cake was washed with cold (5-10 ° C) toluene (2 x 2 times volume) and pumped dry, and then the product was dried under vacuum at 50 ° C. Expected yield: 35% Theoretical value (70% w / w). HPLC (8 minute method) RT 4.30 min. 4-Methenyl-L-benzylalanine ± J ^ fl 27: N-fluorenyl-4-iodo-L-phenylalanine methyl ester (1 times by weight) is suspended in methanol (5 times by volume) and water (5 times by volume) at a concentration of 2 moles Sodium hydroxide (1.73 times by weight) was treated at 22 ° C. (: Under treatment. This mixture was stirred for about 4 hours until the completion of HPLC inspection. The reaction mixture was heated to 31 ± 2 C, then within 20-30 minutes, Add 2 mol hydrochloric acid (0.41 times by weight) at 31 ± 2 ° C, and then seed N- 曱 Si-based-4-erylbenzylalanine (0 · 〇 丨 丨 by weight) at 1: 1 Methanol: Water is added to the slurry. Within 1-2 hours, -37- this or Zhang Fangdu applies the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 200407310 A7 V. Description of the invention (36) ^ 一 ~ ^ Add under 2C Moore 7 ° C hydrochloric acid (1.3 4 times the weight), then check the pH (target pH 1). The white slurry was cooled to 0 ± 3 ° C and then stirred for at least-0 77 4 miles. The solids are collected by the transition to the surface of the cake by pumping the liquid. Consider the cake in 5 soil 5. Was washed with methanol / water (2 times the volume), followed by water (2 x 2 times) at 5 ° C: and then filtered with suction to dryness. The wet solids were dried in a vacuum oven at 30-60 ◦ to produce methyl-4-iodophenylalanine as a white powder. Expected yield: 86-90% of theory; 82-86% weight / weight. HPLC (2 minute method) RT 1.31 minutes 10 fluorenyl-4-ΠΗ-pyrazol-1-yl VL-molybine glutamate j7 interstitial 28: Printed on NMP ( 2.5 times by volume) a solution of fluorenyl-4-iodo-L-phenylalanine (1 times by weight) at 20 ° C with stirring, and aqueous potassium potassium carbonate (1.52 times by weight, 3.5 equivalents). The batch temperature was increased to 40 ° C. Add pyrazole (0.26 times weight, 1.2 equivalents), continue with copper iodide (1) (0.015 times weight, 0.025 equivalents) and racemic anti-1,2-diaminocyclohexane (0.036 times) Weight, 〇. 丨 equivalent). The temperature of this batch was increased to 125 ± 3 ° C and the reaction was stirred for at least 15 hours. The brown suspension was sampled and analyzed by HPLC to determine &lt; 3% SM residue at 20. After cooling to 35 ± 3 ° C, add water (10 times the volume) and continue with DCM (5 times the volume). Activated carbon was added and the mixture was stirred for 1 hour, then the solution was filtered. The phases were separated and the aqueous phase was washed again with DCM (5 volumes). The water phase is cooled to 3 ± 3 ° C, and the concentration is slowly added in about 45 minutes. -38- This paper size applies to China National Standard (CNS) A4 (21 () X 297 mm) Intellectual Property Bureau, Ministry of Economic Affairs Printed clothes for employee consumer cooperatives 200407310 A7 B7 V. Description of the invention (37) HC1 (1.5 times the volume) is acidified and then aged for 15 minutes. Add the remaining HC1 (0.3 times volume) in about 10 minutes to bring the pH to 1. The solid was collected by filtration, and the filter cake was washed with water (4 times the volume) and then replaced with water (4 times the volume) and successively with toluene (3 times the volume). The resulting solid was dried in a vacuum 5 oven at 50-60 ° C. Expected yield: 70-80% of theory, 57-65% w / w. HPLC (2 minute method) RT 1.13 minutes 4-ΠΗ-pyrazol-1-yl VL-benzyl alanine methyl ester hydrochloride 10 Intermediate 29: N-fluorenyl-4- (1Η-pyrazole- 1-yl) phenylalanine (1 times by weight) was suspended in methanol (10 times by volume) and stirred at 22 ± 3 ° C. Acetyl chloride (0,94 times the weight, 3 equivalents) was added dropwise at about 15 minutes at 20-50 ° C. The resulting solution was warmed to 50 ± 5 ° C and maintained for at least 14 hours and 15 hours. The mixture was analyzed. When completed (ie: &lt; 3% amino acid), the solution was cooled at 22 ± 3 ° C and concentrated to about 5 times the volume in a vacuum at &lt; 30 ° C. Isopropyl acetate (10 volumes) was added, and the mixture was concentrated to about 5 volumes in vacuo at &lt; 30 ° C. Isopropyl acetate (10 volumes) was added, and the mixture was concentrated to about 5 volumes in vacuo at &lt; 30 ° C. Then 20 isopropyl acetate (10 volumes) was added and the mixture was cooled to 0-5 ° C. The product was collected by filtration as an off-white solid, washed with isopropyl acetate (2 × 3 volumes), and dried in a vacuum oven at 55 ± 5 ° C. Expected yield: 85-95% of theory, 92-103% weight / weight. HPLC (2-minute method) RT 1.03 minutes. -39- This paper size applies to China National Standard (CNS) A4 (210x 297 mm)

200407310 A7 B7 38 V. Description of the invention 4 NMR (d6-DMS〇) 3.15, 3.23 (ΑΒχ, J 6, 7 Hz, 2H): 3-7〇 (s, 3Η), 4.32 (t, J 6 Hz , 1Η), 6.55 (dd, J 2, 2.5 Hz, 1) H), 7.38 (d, J 8.5 Hz, 2H), 7.74 (d, J 2 Hz, 1H), 7.81 (d, J 8 · 5 Hz, 2H), 8.50 (d, J 2.5 Hz, 1H), 8.70 (br s, 3H). ^ I2SV2-I (ethylsulfanylthio) fluorenyl, a butylsulfanylpyrazole-1. Benzylalanine methyl ester ± MM3:

Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs to prepare acid chlorides (Method 1): (S) -2-Ethylthiomethyl-3-fluorenyl butyric acid in toluene (10 times the volume) ( +) _ Ephedrine salt (1.31 times by weight) suspension, treated with aqueous hydrochloric acid (1 mole concentration, 6.6 times by volume), and stirred vigorously for 15 minutes. The phases were separated and the (upper) organic layer was washed with water (5 times the volume). The phases were separated and the organic layer was concentrated to 2.9 times the volume in vacuo. The solution was treated with thionyl chloride (0.29 times the volume, u equivalent) and was warmed to about 40 ° C for 3 hours. The mixture was sampled (1 drop of 20 dissolved in MeOH (1 ml), aged for 20 minutes, and analyzed by HPLC (2 minutes), RT: 1.33 minutes of acid, 1.61 minutes of Me ester / benzyl). The acid gas solution was cooled to 20-25 ° C and used directly in the next step. Preparation of acid chloride (Method 2): -40- 200407310 A7 B7 V. Description of the invention (39) (S) -2-Ethylthiomethyl-3 -methyl in isooctane (10 ml) A solution of butyric acid (3.75 g) was treated with thionine (1.6 ml) and warmed to about 40 ° C for 3 hours. The mixture was sampled (1 drop dissolved in MeOH (1 mL), aged for 20 minutes, and analyzed by HPLC (2 minutes), rt: 1.33 5 minutes acid, 1.61 minutes methyl ester). The acid chloride solution was cooled to 20-25 ° C and used directly in the next step. Amines coupling (Method 1): 4- (1Η-4Wa-1-yl) -L-phenylalanine methyl S hydrochloride (1 times by weight) printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 10 Suspended in ethyl acetate (5 times the volume) and treated at a concentration of 2 mol I <2C03 (10 times the volume), then stirred at 20-25 ° C until all the solids are dissolved (south to One hour). The phases were separated and the lower aqueous layer was extracted with ethyl acetate (5 times the volume). The combined organic layer is treated on activated carbon (0.25 times weight) at 20-25 ° C for about 2 hours. The activated carbon was removed by filtration, and the activated carbon bed was washed with ethyl acetate (3 volumes). The filtrate was treated with potassium carbonate (2 mole concentration, 10 times volume), and the rapidly stirred biphasic solution was treated with the fbenzene solution of the acid chloride (prepared above) within 20 minutes, keeping the temperature below 25 ° C . The acid chloride was rinsed with toluene (0.1 times volume). The solution was quickly stirred for 30 minutes and the phases were separated. The upper organic layer was washed with water (5 times the volume) and then concentrated to 8 times the volume by atmospheric distillation. The solution was maintained at 75-80 ° C, and slowly diluted with 2,2,4-trimethylpentane (TMP, isoxine, 16 times volume) within 30 minutes, and maintained at 70-80 ° C15 minutes to allow crystallization to develop. The thin slurry was cooled to 0-5 ° C in at least 2 hours. The solids were collected by filtration, cold (5 ° C) ethyl acetate / butyl MP (1 ·· 3, 4 times the volume), -41- This paper is suitable for Chinese National Standard (CNS) A4 specifications (210 · \ 297 Public Love) 200407310 B7 A7

It was then washed with TMP (4 volumes) and dried in a 55r vacuum oven to produce the title compound as an off-white solid. Expected yield: 74. /. Theoretical value, 11% weight / weight. HPLC (2 minutes method) RT 1.64 minutes 5 amidine coupling (Method 2): 4- (1Η-Η 峻 小 基) _L-phenylalanine ammonium ester hydrochloride printed by the Consumer Cooperative of Intellectual Property Bureau of the Ministry of Economic Affairs (50 g) was freshly floated in ethyl acetate (75 ml) and treated with 2 mole K2CO3 (33 ml), and then stirred at 20-25 C until all solids were dissolved and dissolved. . The rapidly stirred biphasic solution was treated with a solution of the acid chloride (prepared above) within 20 minutes, keeping the temperature below 25t :. The solution was quickly stirred = 30 minutes and the phases were separated. The upper organic layer was washed with molar concentration Ηα (33 ml) and water (33 ml), and then concentrated to 35 liters by atmospheric distillation. The solution was maintained at 75-80 C &apos; and slowly diluted with 2,2,4-trimethoprim 15 pentamidine (TMP, isooctyl, 85 ml) over 30 minutes and maintained at 70-8. 0 15 minutes' to allow crystallization to develop. The thin slurry was cooled to 0-5 ° C in at least 2 hours. The solid was collected by filtration, washed with cold (5.0) ethyl acetate / TMP (1: 4, 25 ml), then cold TMP (30 ml), dried in a vacuum phase at 55 ° C, and produced as off-white The title compound as a solid, 6 4 20 g, 87% yield. HPLC (2 minute method) RT 1.64 minutes Example: -42- This paper size is applicable to Chinese National Standard (CNS) A4 (210x297 mm) Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs Consumer Cooperatives 200407310 A7 B7 V. Description of the invention (41 )

Paste example 1: N-「(2S) -2- | Y 输 基基) _3_ 曱 基丁丁基 外, sitting on the small screen ντ. · ^ 10 Some alanine (Method 1) AM (2S) -2-[(Ethylthio) fluorenyl] -3-fluorenylbutanyl 4- (1Η_πpyrazole small group) cardinyl alanine methyl ester (8.17 g, 19.6 mmol) was dissolved in In a mixture of THF (75 liters) and methanol (75 ml), it was deoxygenated for 40 minutes under a stream of nitrogen. Similar to degassed 2 Molar sodium hydroxide (98 ml, 196 15 mmol) at 0 ° C. The solution was added dropwise. The reaction was stirred for 20 minutes at 0.0, and then for 2 hours at room temperature. The mixture was re-cooled to 0 ° C, and the mixture was acidified with 5 mol hydrochloric acid (42 mL) and at It was distributed between ethyl acetate (800 ml) and water (400 ml). The organic phase was separated, washed with brine (400 ml), dried over NaAO4, and the solvent was removed to give a crude product of 20 yields. This was purified with Biotage and was eluted with a gradient of 1% to 3% methanol in dichloromethane containing 0.5% acetic acid to produce 6.67 g of pure material. LCMS RT 2.98 minutes MH + 362, Mi [(2SV2 · “(fluorenyl曱 基 • 曱 基丁 醯 some mouth stupid -43- paper rule Applicable Chinese National Standard (CNS) A4 size (210x297 mm)

200407310 A7 B7 V. Description of the invention (42 Alanine (Method 2) 10 15 Printed by the Consumer Consumption Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 20 A /-((2S) -2-[(Ethylthiomethyl) -3-methylbutanyl] · 4 · (〗 Η-α 比 °°-丨 .- 基) 丄 -phenylalanine methyl § intent (33.5 g) was fed into the container and vacuumed 3 times / Nitrogen cycle deoxidation. The solid was suspended in a mixture of water (3 times the volume, chuan 0 liters) and MeOH (8 times the volume, 270 ml), and fell off under (3, rat air. At 10 Add 32% weight / weight (108 moles) NaOH (j.5 equivalent '0.78 times volume, 26 strokes) in minutes, and mix the mixture at 20 C for 60 minutes. Once started The starting material was consumed, tributylphosphine (0.02 equivalents' 0.012 times volume, 0.4 liters) was added, and the solution was stirred for 60 minutes. The solution was filtered into a new reaction vessel with a liner, and water (3 times the volume,丨 〇〇 litre) liner washing. The clear solution was acidified to pH 4.9 by adding 36% w / w (11.6 mole concentration) HC1 (2.7 equivalents, 0.57 times volume, 19 ml) within 15 minutes, and 15 points aging Bell to allow the crystallization to develop. Add 36% w / w (11.6 mole concentration late (: 1 (1.9 equivalents, 0.39 times volume, 13 ml)) to the pulp to acidify it to pH 1.2. The pulp is cooled to 0-5 ° C, and aged for 30 minutes, then the solid was collected by filtration, washed with cold (0-5 ° C) 1: 1 MeOH / water (2 x 3 volumes, 100 ml), and vacuumed at 50 ° C Drying in an oven yielded the title compound as a gray-yellow to brown powder, 28.04 g, 96.7% yield. Prepared similarly: Example 2: NK (2S) -2- (sulfofluorenyl) -3 -a certain butylfluorenyl 1 -4-Pyrimidine-5-yl alanine-44- This paper size is applicable to Chinese National Standard (CNS) A4 (210 X 297 public love) 200407310 A7 B7 V. Description of the invention (43) From 丨 (2S ) -2-[(Ethylthio) methyl] -3-methylbutylfluorenyl} -4-pyrimidin-5-yl) -L-benzylalanine methyl ester LCMS RT 2.66 min MH + 373 5 Examples 3: Nf (2SV2-f (mercaptofluorenyl V4-fluorenylpentylfluorenyl 1-4-ΠΗ-roazol-5-yl VL-benzyl propionate) from # -U2S) -2-[(乙醯Methylthio) methyl] -4-methylpentanyl} -4- (1 (-pyrazol-5-yl) -L-phenylalanine methyl ester 10 LCMS RT 2.94 Minutes, MH + 376 Example 4: N-"(2SV2-" (g-Alkylfluorenyl V3-fluorenylbutylfluorenyl1-4-ΠΗ-pyrazol-5-yl) -L-benzylalanine I 萣 15 from / vSf (2S) -2-[(Ethylthio) methyl] -3-methylbutylfluorenyl} 4- (1fluorene-pyrazol-5-yl) -L-phenylalanine methyl ester LCMS RT 2.79 Minutes MH + 362 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 5: 20 N-`` (2S) -2-f (mercaptofluorenyl) -3-fluorenylbutanyl 1-4- (1Η-pyrazole-4- 1-L-benzylalanine from iV-U2S) -2-[(ethylfluorenylthio) methyl] -3-fluorenylbutanyl} -4- (1fluorenyl-pyrazol-4-yl)- L-phenylalanine methyl ester LCMS RT 2.72 minutes, MH + 362 -45- This paper size applies to China National Standard (CNS) A4 (2 Sichuan X 2 (J7 mm) Printed by the Consumer Cooperative of Intellectual Property Bureau of the Ministry of Economic Affairs 20040710 A7 B7 V. Description of the invention (44) Another process of Example 1: yV] (2S) -2-[(ethylamidothio) methyl] -3-amidobutanoylpyrazol-1-yl) -L-phenylalanine phosphonium ester (1 times by weight) was fed into the vessel and deoxygenated in 3 vacuum / nitrogen cycles. The solid was suspended in a mixture of water (3 times the volume) and MeOH (8 times the volume) and stirred at 20 ° C under nitrogen. 32% w / w (10.8 mole concentration) NaOH (3.5 equivalents, 0.78 times volume) was added over 5 minutes, and the mixture was stirred at 20 ° C for 60 minutes. 10 Once all the starting material is consumed, tributylphosphine (0.02 equivalent, 0.012 volume) is added and the solution is stirred for 30-60 minutes. The samples were analyzed by HPLC to determine that most of the disulfide was reduced back to the starting material. The solution was filtered into a new reaction vessel with a liner, and the strips were washed with a water (3-fold volume) liner. The clear solution was triturated to a pH of 3.0-3.5 with the addition of 36% w / w (11.6 mole 15 concentration) HC1 (3.4 equivalents, 0.70 times the volume) g, and aged for 10 minutes to allow crystallization to develop. The slurry was acidified to pH 1 by adding 36% w / w (1 1.6 mole concentration) HC1 (1.2 equivalents, 0.24 times volume). The slurry was cooled to 0-5 ° C. And aged for 30 minutes, then the solids were collected by filtration, washed with cold (0-5 ° C) 1: 1 MeOH / water (2 x 3 volumes), and at 50 Drying in a vacuum oven at 20 ° C yields N-[(2S) -2-[(mercaptomethyl) -3-fluorenylbutyryl] -4-(1Η-° ratio σ sitting -1 -yl) -L-benzyl propionic acid 'is a gray to white powder. Expected yield: 92-95% of theory, 80-82% weight / weight. HPLC (2 minute method) RT 1.45 minutes. -46- This paper size applies to China National Standard (CNS) A4 (210 x 297 mm)

Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 200407310 A7 B7 V. Description of the Invention (M). 4 NMR (d6-DMS〇) 0.81, 0.86 (2d, J 7 Hz, 6H), 1.58 (dd, J 7, 9 Hertz, 1H), 1.72 (eight peaks, J 7 Hertz, 1H), 2.15 (M, 1H), 2.42-2.59 (m, 2H), 2.94 (ABX, J 10, 14 Hertz, 1H), 3.11, 3.14 ( ABX, J 5, 14 Hz, 1H), 4.59 (m, 1H), 6.51 (t, J 2 Hz, 5 1H), 7.38 (d, J 8 Hz, 2H), 7.72 (m, 3H), 8.26 ( d, J 8 Hz, 1H), 8.43 (d, J 2 Hz, 1H), 12.7 (br s, 1H). Analytical method 8 minutes Method parameter numerical analysis column 50 mm x 2.0 mm LunaC18 (2), 3 micron mobile phase mobile phase A: 0.05% vol / vol in TFA solution mobile phase B: 0.05% vol in MeCN / Volume TFA solution gradient time (minutes) mobile phase A (%) mobile phase B (%) 0.00 100 0 8.00 5 95 8.01 100 0 10.00 100 0 flow rate 1.0 ml / min temperature 40 ° C UV detection at 220 nm Injection volume 1 microliter -47- This paper size is applicable to China National Standard (CNS) A4 (210 x 297 mm)

200407310 A7 D / Description of the invention (46) Approximate time --------- 10 minutes 3-minutes! ^ Method parameters --__ Numerical analysis tube; (: main 50mm x 2.0mm Luna C18 (2), 3 micron mobile phase mobile phase A: 0.05% volume / volume TFA solution in water mobile phase B: in 0.05% volume / volume TFA solution gradient time in MeCN (minutes) 0.00 2.00 2.01 3.00 mobile phase A (%) 100 100 100 mobile phase B (%) 0 95 0 0 flow rate 2.0 ml / min temperature *-~ --- -—. 60t UV detection, injection volume of 1 microliter at 220 nanometers, about 3 minutes of time. 5 The printing of LCMS information of the consumer cooperative of employees of the Intellectual Property Bureau of the Ministry of Economic Affairs, the following characteristics are generated on the system: column: 3.3 cm x 4 .6mm inner diameter, 3 micron ABZ + PLUS Flow rate: 3 liters / min Injection volume: 5 microliters -48- This paper size applies to China National Standard (CNS) A4 (210 X 2 () 7 mm) 200407310 A 7 B7 V. Description of the invention (47 Temperature: RT UV detection range: 215 to 330 nanometers) Capacitor: A: 0.1% formic acid + 10 ml molar concentration of acetic acid recorded B55% acetonitrile + 0.05% formic acid gradient: time ( Minutes) A (%) B (%) 0.00 100 0 0.70 100 〇4.20 〇100 5.30 〇100 5.50 100 0 ίο Ministry of Economic Affairs Printed by the Employees ’Cooperative of the Intellectual Property Bureau 15 Biological data Anti-ACE inhibitory activity The anti-ACE inhibitory activity was measured by the following analysis method using the matrix MCA-Ala-Ser-Asp-Lys-Dpa-ΟΗ 320 μm excitation / 400 nm increased fluorescence. 1 μl of test compound solution plus TCEP (1: 2.5 compound: TCEP) in acetone / water (丨: I), and 15 μl of matrix Solution (176 microa ear concentration in 10 micromolar TCEP solution) and 15 microliters of human ACE in buffer (200 picomolar concentration at 50 millimolar concentration HEpes pH 7.4, L0 mol molar concentration NaCi solution, micromolar zinc acetate, using 1 equivalent NaOH, pH to 7.4) and 10 micromole TCEP (final concentration of about 100 picomolar human Α (: ε and / continued) Degree 88 -49- This paper size applies Chinese National Standard (CNS) A4 specification (2 丨 () X 297 public: 200407310 A7 B7 V. Description of the invention (48) MCA matrix with micromolar concentration) mixed in 60 minutes After incubation, use a Tecan SpectraFluor Ultra fluorescence plate reader or equivalent and incubate at 320 nm. / 400 nm emission emit fluorescence readout. Note: Human ACE means human kidney ACE. For human plasma analysis and 5 mouse plasma analysis, the analysis used was the same as above, except that human / mouse plasma replaced buffered human ACE. The method for analyzing the inhibitory activity against rabbit NEP was performed as described above with the following changes. Recombinant rabbit kidney NEP 10 replaces human ACE and uses N-dansyl-D-Ala-Gly-p nitroPhe-Gly as a matrix instead of MCA-Ala-Ser-Asp-Lys-Dpa-. H. The biological data (ρKι) for the selected compounds are shown below (compounds are identified as above, except for compound X). 15 (〇 Intellectual Property Bureau of the Ministry of Economic Affairs, Consumer Cooperatives, printed compounds, human ACE ρκι, human plasma ACE ρκι, mouse plasma ACE ρκι, rabbit NEP ρκι 1 9.0 7.1 7.1 &gt; 8.5 2 8.1 6.9 6.1 &gt; 8.5 〇j 9.2 6.9 7.1 &gt; 8.5 4 9.0 7.4 7.2 &gt; 8.5 5 8.6 7.1 6.9 &gt; 8.5 -50- This paper size applies to China National Standard (CNS) A4 (2U) X 297 mm) 200407310 A7 B7 V. Description of the invention 49 X 8.0 5.1 5.8 8 , 5--&gt; 8 ^ &gt; 8.5 (Π) Compound human ACE ρκι human plasma ACE ρκι _ --- ^ mouse m-poly ACE ρκι rabbit NEP ρκι 1 8.8 7.0 ....... _ a ~ 7.1 8.8 2 8.1 6.9 ---- ^ 6.1 &gt; 8.5 J 9.2 6.9 ----- 7.1 &gt; 8.5 4 9.0 7.4 Circle ----- 7.2 &gt; 8.5 5 8.6 7.1 6.9 &gt; 8.5 X 7.8 5.2 5.8 9.1 Y * 8.5-......- 1 -------- &gt; 8.5 compound 2-benzylmethylpropanyl) _4- (2-σsedino) -phenylalanine compound benzene (Methylpropanyl) -4- (5- ° dense bite) _Phenylpropanine printed by the Intellectual Property Bureau of the Ministry of Economic Affairs Employee Consumption Cooperative Society Note: Compounds X and Y can be prepared according to the process provided by World Patent W097 / 24342ο Tables ⑴ and (11) represent the values obtained from repeated tests. The compound (1-5) of the present invention shows increased human ACE ρκι, human blood ACE pKl, and mouse plasma ACE team when compared with compound X "This -51-This paper is in accordance with the Chinese National Standard (CNS) A4 specification (210 x 297 mm) 200407310 A7 __________ B7 V. Description of the invention (50) Surprising efficacy refers to the improved ace_nep inhibitory activity. The main compound can be tested for ACE inhibitory activity and used for angiotensin inhibition Test of α ACE-mediated angiotensin {converted to angiotensin II, measured using purified human ACE enzymes. The ability of the compounds of the present invention to inhibit this conversion is from The change ratio is calculated in accordance with the print. 1 2 5 Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs, Consumer Cooperatives. This paper size applies to China National Standard (CNS) A4 (210 X 297 mm).

Claims (1)

A B c D 200407310 6. Scope of patent application 1. A compound of formula (I): Wherein: R1 represents Cw alkyl; 5 R2 represents pyrazole or pyrimidine; or a pharmaceutically acceptable derivative thereof. 2. The compound according to item 1 of the scope of patent application, wherein R1 represents a Cm alkyl group. 3. The compound according to item 1 of the scope of patent application, wherein R1 represents isopropyl 10 group. 4. A compound according to any one of claims 1 to 3, wherein R2 represents pyrazole. 5. The compound according to item 1 of the scope of patent application, which is selected from N-[(2S) -2-[(mercaptofluorenyl) -3-fluorenylbutylfluorenyl] -4- (1fluorene-pyrazol-1-yl) benzene 15-Alanine printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs; N-[(2S) -2-[(fluorenylfluorenyl) -3-fluorenylbutanyl] -4-pyrimidin-5-yl-L- Phenylalanine; N-[(2S) -2-[(S1l-methylfluorenyl) -4-fluorenylpentanyl] -4- (1fluorene-pyrazol-5-yl: HL-phenylalanine Acid; 20 N-[(2S) -2-[(mercaptofluorenyl) -3-fluorenylbutylfluorenyl] _4- (1fluorene-pyrazol-5-yl) -L-benzene-53-This paper is applicable to China Standard (CNS) A4 specification (210x297 mm) 200407310 A8 B8 C8 D8 Patent application scope ο 15 propyl alanine; N-[(2S) -2-[(mercaptomethyl) -3-methylbutylfluorenyl] -4 -(1'-pyrazol-4-yl) phenylalanine; and a pharmaceutically acceptable derivative thereof. 6. A compound according to any one of claims 1 to 5 of the scope of patent application for treatment. 7. -A pharmaceutical composition comprising a compound according to any one of claims 1-5 in the scope of the patent application, and a pharmaceutically acceptable carrier and / or excipient thereof. 8. According to any one of claims 1-5 in the scope of patent application Itemization The use of a compound for the manufacture of a medicament for treating a patient suffering from a disease that may be improved with an ACE and / or NEP inhibitor. 9. A method for treating a patient suffering from a disease that may be improved with an ACE and / or NEP inhibitor. , Including the administration of a therapeutically effective amount of a compound in any one of the scope of applied patents. 10.-A method for preparing a compound of formula (I): R2 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs Rl (I) includes reacting a compound of formula (II) with a compound of formula (III) and continuing deprotection: -54-This paper size applies to China National Standard (CNS) A4 (210x297 mm) 200407310 8 8 8 8 AB c D Patent application scope R2 (Π) 〇 ^ s /-^ co2h Cover 1 (III) where: R1 represents a Cm alkyl group; R2 represents a methyl group or a sigma group; and P1 represents an oxygen protecting group 11. A compound of formula (II): R2 (II) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 10 where: R2 stands for σσ or σ 定; and P1 stands for a protecting group -55- This paper size applies the Chinese National Standard (CNS) A4 specification (210x297 mm) 200407310 (1). The designated representative map in this case is: Figure ___ (none) (II). The component representative symbols of this representative map are simply explained: beneficial If there is a chemical formula in this case, please disclose the chemical formula that best shows the characteristics of the invention: Page 2-1