JPH03505875A - Retroviral protease binding peptide - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Retrovirus protease binding peptide background Retroviruses, or viruses of the Retroviridae family, decode their genetic material. It is a member of the virus family that transports ribonucleic acid rather than xyribonucleic acid. Also, R.N. Also known as A tumor virus, its presence is associated with a wide range of diseases in humans and animals. ing. They are Rous sarcoma virus (R3V), feline leukemia virus (ML). V) Mouse mammary tumor virus (MMTV), feline leukemia virus (FELV) ), bovine leukemia virus (BLV), mesogyfizer monkey virus (MPMV), simian sarcoma virus (SSV), simian acquired immunodeficiency syndrome (SSV) AIDS), human T-lymphotrophic virus (HTLV-1, -11), AIDS Human acquired immunodeficiency virus (HII-1, HIV) is the causative agent of AIDS (AIDS). -2), considered to be a causative agent in pathologies associated with AIDS-related complexes and other infections. It is. Although the pathogen has been isolated in many of these cases, this type No effective method has yet been developed to treat this type of infection. Of these viruses , HTLV and HIV are particularly well characterized.

Although they differ in details, all retroviruses are similar in their overall structure. cell The outer virus particle has an outer membrane dotted with viral glycoproteins, a core of structural proteins, and a rib. Consists of a single-stranded getum of nucleic acid. Retrovirus genome is 5° gag T) It has a unique local configuration called 01-env-3', and the geg region hoods the core structural protein, and the pot region contains reverse transcriptase, integra encodes certain critical viral enzymes such as enzymes and proteases, The enV region encodes the outer membrane glycoprotein. Virus replication occurs only within host cells. and depends on host cell function. Critical to this replication are functional viral proteins. production. Protein synthesis is an oven reading frame gag%po! and translation into a polyprotein structure corresponding to the env reading frame. , which is processed, at least in part, by viral proteases into functional proteins. be done. Proteins provided by viral proteases in polyprotein processing Degradation activity cannot be provided by the host, and the life cycle of retroviruses It is essential for Kuru. In fact, retroviruses or their variants that lack proteases have been shown to be non-infectious. Kato et al. ), pyrology (v iro l ogy), 145.280-92 (1 985) See outside. Therefore, inhibition of retroviral proteases provide treatments for viral diseases.

Retroviral proteases are not very well characterized. proteer Both enzymes and their polyprotein substrates are recovered in very low yields from pyrion particles. . In order to evaluate their activity, it is necessary to provide a substrate to express proteolytic activity. It's necessary. The use of natural substrates for proteolytic activity assays is important for substrate production and purification. Many of the shortcomings of quantification and quantification (the use of small, synthetically produced and purified peptides) It can be overcome with use. Therefore, small molecules that can be used as substrates for these enzymes The production of unique peptides is highly desirable and advantageous.

Proteases are enzymes that cleave peptide bonds in proteins and polypeptides.

They are present in most biological systems and serve both metabolic and regulatory functions. . Their role in metabolism transforms polypeptides into smaller peptides, ultimately It is hydrolyzed into its constituent amino acids. Their role in biological regulation It is widely known and has been the subject of several reports. For example, H. Edited by Lutzer et al., Biological Functions of Proteases, (19 See 79).

One of their regulatory roles is the post-translational processing of polypeptides to produce functional proteins. , production of enzymes and mesoieters. Essential for their regulatory functions is polymorphism. It is the ability to act selectively on peptides. This selectivity consists of two factors. ing. ■) the ability to act on specific substrates, and 2) the ability to act only on specific peptide bonds. It is the ability to hydrolyze. At the structural level, this substrate specificity is determined by the primary amino acid sequence. sequence, the result of the local structure of the substrate polypeptide and the peptide bond at the cleavage site. This is the result of the amino acids that form.

in polyprotein natural substrates for some retroviral proteases. Sequence information regarding putative cleavage sites is available. However, retroviruses - No peptide substrates for proteases are known so far. Also, heaven Since the natural substrate binding interactions are unknown, it is difficult to determine which length peptides are accepted. is often difficult to predict. Similarly, the structure of a large polypeptide is which in small peptides can be significantly different than those in small peptides. It is difficult to predict which amino acid sequences will bind optimally.

Retroviral protease testing by amino acid sequencing of viral proteins suggests certain common amino acid residues in the cleavage region for. for example , Casey et al., J., Viro 1. , 55.417-23 (1 ,985) i;LHIV-1 p24 protein amino terminal sequence Pr o-I 1e-Va 1-Gln-Asn, and this cleavage site contains This indicates that a proline residue is included. Jimarzo Vuelones (d iMa rzo Veronese) et al., 5science, 231. 12 89-90 (1986) is a reverse transcriptase (RT) cleavage site (* ) is proposed as Thr-Leu-Asn-Phe*-Pro, and RT is given a proline/°r mino terminus. Debouck et al., Pr. oc, Nat 1. Acad, Sci, USA, 84°8903-06 (1 (987) suggested that the HIV-1 protease is autocatalytic; The amino terminal of the protease has the sequence Pro-Gln-11e-Thr-Leu. It is disclosed that it contains. Peari et al., Nature, 328, 482 (1987) describes certain core precursors for a large group of retroviruses. Comparing polypeptide substrates, X-Pr for retroviral proteases Suggests sequence selection to cleave at o. Here, X usually represents an aromatic amino acid (P he, Tyr) or large, hydrophobic amino acids (Met, Leu). Sara This sequence is usually separated by a small, hydrophobic amino acid on either side. This suggests that the

Structures of many retroviral proteases, primarily their nucleotide structures. tested based on the predicted amino acid sequence deduced from They are generally po the 5° end of the t region, the 3' end of the gag region, or between these two regions (however, coded outside the frame). Yasunaga et al., FEBS Lett, 199.2, 145-49 (1986). We disclose sequence homology between Lentiviridae and Retrovirus I (TLV/ Some highly conserved sequences within the BLV subfamily are shown.

Deborah has developed a method for expressing retroviral proteases in E. coli. (Debouck) et al., Proc, Nat 1. Acad, Sci, USA, 8903-06 (1987) and Graves for the HIV-1 virus. (Graves) et al., Proc.

Nat 1. Acad, Sci, USA, 85.2449-53 (1988 ).

To date, little biochemical characterization of retroviral proteases has been performed. not present. However, Katoh et al. It has been suggested that it may be a protease. This is an acid protease inhibitor. Retroviruses associated with pepstatin BLV, MMLV and HTLV Based on the observation that it has been shown to be a weak inhibitor of S. proteases It is something that can be developed.

U.S. patent for method for producing protease inhibitors of retroviral proteases Nos. 4,636,492 and 4,644,055. According to this, the professional Tri- and tetrapeptides that substantially correspond to the cleavage site amino acid sequences of the tease substrates. The peptide is modified to have a C-terminal halomethylketone group. halomethylke ton is a reactive group, and once the peptide is bound, it reacts with the protease. Conceivable.

Although it is not related to retrovirus protease, Umezawara et al. ci, Soc, Press, Proteinase Inhibitors, pp 3-15 (1983)) screened microbial culture filtrates to identify various groups. We have isolated inhibitors of proteases. Some of these peptide inhibitors , for example, bestatin, amastatin and pepstatin are not d-amino acids Contains special amino acids. These inhibitors inhibit proteolytic enzymes at the cleavage site. It is said that it functions by the presence of a non-hydrolyzable C-C bond. Ru.

Pepsta, a pentapeptide inhibitor isolated from Streptomyces cultures Chin has been shown to broadly inhibit acid proteases, including renin and pepsin. It is attracting attention due to its ability to inhibit (IJr+1ezawa) et al., J. Ant 1biot, 23.259-263 (1970)). Pe Pustatin is a special amino acid called statin, (3S,4S)-4-amino acid. No-3-hydroxy-6-methylhebutanoic acid was added to the pentapeptide (AHMHA). Contains in two positions. This special amino acid is not a d-amino acid, but a dipepeptid. It has a tide-like function, and instead of the cleaved peptide bond, the C-C bond attached to the enzyme is It is considered to give. Peptide derivatives contained in statins are human proteases , has been used to inhibit renin and pepsin (US Pat. No. 4,481,192).

Several groups have proposed modifications that optimize the renin-inhibiting activity of hefstatin. It is being done. Ricc et al., J. Med.

Chem, 23.27-33 (1980) describes statin combinations with pepstatin. showed that the chirality of statins is important for binding, and modified statins to improve their structure and inhibitory activity. Exploring sexual relationships. Thus, (3S.

4S)-4-amino-3-hydroxy-5-phenylpentanoic acid (AHPPA) When combined with a peptide instead of a statin, it can have the same effect as a statin. It has been proven.

On the other hand, other [dipeptide analogues] containing a C-C bond instead of a peptide bond It has been synthesized and studied by several groups.

Many of these attempts involve modification of carbons with hydroxyl groups and adjacent carbons. Contains a structure similar to that of statins. For example, oxo derivatives of statins (J, Me d, Chem, -, 30, 976-82 (1987)), deshydroxy derivatives (J, Med, Chem, 23.27-33 (1980)'), phosphorus-containing derivatives (J, Med, Chem, 1603-09 (1987)) and 7/ lz oro derivative (J.

Med, Chem, 30, 1617-22 (1987); J, Med, C hem, 20.2080-87 (1986)) was synthesized and its renin and Pepsin inhibitory ability has been reported. against the carboxyl group of statins A variant with a substituted methylene group inserted at alpha has also been reported to inhibit 1/nin ( EP-A229667; PCT/WO38105050: US Patent 46650 55: EP-A173481). These “dipeptide analogs” They show disparate activity in inhibiting pepsin and pepsin, and their specific activity is linked to binding to the enzyme This indicates that it depends on the neighboring group to convey the . Employed in these disclosures The synthetic methods described herein are incorporated herein by reference.

Assays protease activity and inhibits retrovirus protease activity for pharmaceutical use. A peptide that binds to retroviral proteases is required to provide a harmful agent. It is considered essential. Such medicinal uses provide treatment for previously untreatable diseases. do.

Summary of the invention The present invention provides a protein that binds to a retrovirus protease represented by the formula (I) below. It consists of peptides. In one respect, these peptides can be assayed for protease activity. It is useful as a substrate for In other respects, these peptides It is an inhibitor of enzymes and is useful in the treatment of diseases related to infections with these substances.

The present invention also provides a pharmaceutical composition comprising a compound of formula (1) and a pharmaceutically acceptable carrier. It is a product.

The present invention further provides administering an effective amount of a compound of formula (1) to a mammal in need of treatment. It constitutes a treatment for viral diseases consisting of:

The invention also provides assays for viral proteases.

Furthermore, in another aspect, the present invention provides for use as an intermediate for producing the peptide of formula (1). after making the peptide resistant to degradation by viral proteases. It is a compound of formula (Ilb) shown below.

Detailed description of the invention The peptide of the present invention has formula (I).

A, -B-(Q)a-(C)b-(D)c-M-(W)d-(X)e-Y-Z The compounds shown and their pharmaceutically acceptable salts.

In the formula, A is BocNHSCbzNH, H, R'R″N, R″C0NR″ or D nsNH or when a, , b and C are O and Y is a covalent bond, A is H, Bo cSCbz, R'' or R''CO.

B is one or more D or L-amino acids, β-Ala or a covalent bond.

C and D are the same or different, GlxXAsx, Ala, β-AlaSA rg, Gly, I le, Leu, Lys, Ser, Thr, Va I, Me t or His.

Q is a D or L-amino acid, Ser, Thr, Asp, His.

Cys, Arg or Ala.

W is Pro or Δ3-dehydroPro.

X is Ala, Gly, lie, Leu, Val, Met, Lys.

Glx or Asx.

Y is one or more D or L-amino acids or a covalent bond.

Z is Co, R"", C0NR' R"", COR', CH, OH, CH, NR’ R″” or H or when d and e are O and Y is a covalent bond, Z is OR"" or NR'R"".

a, b, c, d and e are each independently O or 1, but C and and e are not O at the same time.

M is Cha, Phe (4'R,) or -NHCHR,R,-.

Here, R1 is independently Cl-5A Ik, (CH,)nSC,, A1 k.

(CH,)nOcI-5A]k, (CH2)ncs-77 chloroalkyl or ( cH,) mcaHa-Ra.

R6 is halogen, OR', No, NH, or H.

R2 is (C)! 2), +, CHR3(CH2)IT, Co+, Co(CH2)rn Co+.

CHR3CHR3I(CH2)mCo-, CHR3CHR3ICHRl (CH 2 deception Co-.

CHR3CHRICHR' (CH21mCo-, CHR3CHR'CHR1( CH21, nCo+.

C0CHR4CHR'(CH2)ntCo-, C0CF2CR'R"(CH2) mco-, CO (J2(C)121mCO-, R2 to CH-CR'CHR'+) rr, Co-, C0CH-, CR'(CH2)mCo-.

R5 and F3' are each independently OH, H or NH.

R4 is OH, H, NH, or -0.

m is independently 0, l, 2 or 3.

n is independently 1 or 2.

p is 0, 1 or 2.

R″ is H or C, , Alk.

R'' is H or C,-, AIk.

R” is HlC, -, Alk SC, -s Schiff 0 Furkyl, (CHt) ncs Hs, (CH,)nCsH,N, (CH,)nOH, (CH,)nNH,t, is (CH,)nNHC(NH)NH,: It is.

The invention also includes pharmaceutically acceptable addition salts, complexes or salts of the compounds of the invention. Also includes Rotoragu. Protorag is inviv.

any covalently bonded carrier that releases an active crude compound of formula (I) in it is conceivable that. When different from the usual notation, A in formula (I) is the terminal end of the peptide. contains an amine group, and Z contains the terminal carboxy group of the peptide. A and When Z is H, the terminal residues of the peptide are ``des-amino'' and ``descal'', respectively. boxy” amino acid.

Using this expression, either A contains the terminal amine 7 group of the residue corresponding to B, or B shares In the case of a bond, it corresponds to Q. Alternatively, if a is O and B is a covalent bond, then to C, B If is a covalent bond and a and b are O, it corresponds to D. B is a covalent bond, and a, b and and when C is O, the amine 7 group of M is the atom specially given by A in formula (1). Substituted with syl or alkyl groups. Similarly, Z is the amino acid residue corresponding to Y. contains a terminal carboxyl group, or if Y is a covalent bond, to X or if Y is a covalent bond. When d and e are 0, the terminal carboxyl group of M is specially given in formula (1). will be replaced by Z.

By manipulating the subscripts a to c of formula (1), the usefulness of the compound is maintained and the peptide is It shall be indicated that residues may be removed from the amino terminus. Such operations are unless Y is a covalent bond, a is not O, and a b is not 0 unless is O, and if b is O, then C is not O. carboxyl At the proximal end, e is not O unless Y is covalently bonded. More details below As described, d works independently, and when M is Cha or Phe (R,) , 1, M or -NHCHR, R2, 1 or C1 is preferably 0.

Other abbreviations and symbols used herein to represent peptides are common in the art. It is commonly used.

Amino acid 3 letter code 1 letter code Asparagine Asn Aspartic acid N Aspartic acid Asp D-Methionine Met Met M Phenylalanine Phe F Proline 　 　 　 　 　 　 　 　 　 in Q glutamic acid Glu E-Glycine Cry Cry G-His Tamin His isoloinon Ile I Asparagine or Asx B aspartic acid Glutamine or Glx Z-tryptophan According to the conventional expression method, the amine end is left, carboxy terminus on right. Unless otherwise specified, all xyl amino acids (A A) is assumed to be of L absolute configuration. (4' R,) Phe is the 4th position of the phenyl ring represents phenylalanine substituted by R6. If R6 is H, the residue is It constitutes nilalanine, and when R, is ○H, the residue constitutes tyrosine. β-A la represents 3-aminopropanoic acid. Cha represents ncrohexylalanine.

BoC is a t-butyloxyphosphoryl group, DnS is a dansyl group, i.e. -dimethylaminonaphthalene-5-sulfonyl, CbZ is carbobenzyloxy group, BrZ is an 0-bromobenzyloxycarbonyl group, C1z is a p-chlorocarbonyl group, and C1z is a p-chlorocarbonyl group. Bobenzyloxy group C1, Z is 2,4-dichlorocarbobenzyloxy group, B zl is a benzyl group, MeBzl is a 4-methylbenyl group, Ac is acetyl, Alk Or C,, Alk is CI-, alkyl, Ph is phenyl, DCC is dicyclo xylcarbodiimide, DMAP is dimethylaminopyridine, HOBT is 1-hydroxy Droxybenzotriazole, NMM is N-methylmorpholine, DTT is dithio Threatol, EDTA is ethylenediaminetetraacetate, DIEA is diisopropyl ethylamine, DBU is 1,8-diazobicyclo[5,4,0]undec- 7-ene, DMSO is dimethylformamide, DMF is dimethylformamide or and THF represent tetrahydrofurano. HF is hydrofluoric acid and TFA is trifluoride. Indicates lifluoroacetic acid. In this specification, C I-, alkyl include methyl, ethyl l, propyl, isopropyl, butyl, isobutyl, 5ec-butyl, t-butyl It includes pentyl, pentyl, and isopentyl. The carbon used in the compounds of the invention Asp and Glu with sulfonic acid side chains have free carboxylic acid, Cl-5 alkyl Includes kill and benzyl ester side chains. C, , A l k is 1 to 18 It includes both straight chain and branched chain alkyl having carbon atoms.

The peptides of the invention interact with viral proteases in a manner that mimics the binding of natural substrates. Join. The peptide is generally a dodecapeptide or smaller. death However, it contains a residue defined as -Q-C-D-M-W-X- as shown in formula (1). Longer peptides are also considered active and are within the scope of the invention. Promotion of peptides The residues closest to the fixed cleavage site, denoted M-W, are critical for binding. . X is preferably a neutral or acidic amino acid. Preferably, X is Ala, Gl y, Ile, Leu, Val, Met, Lys, Glx or Asx . Valine is particularly suitable. D is preferably a neutral, hydrophobic amino acid, or A It may also be certain hydrophilic amino acid residues such as sp and Set. Gln,As n, Ala, β-Ala, Ile, Leu, Val and Met are preferred. Yes. Particularly preferred are Gln, Asn and Ala. The residue corresponding to C is neutral, Either acidic or basic amino acids may be used. Glu, Gln, Arg, Lys , Ser, Ala, β-AIa, , Asn and cry are preferred. Gl n, Asn and Ala are particularly preferred as C. Residue Q is D- or L-3e Hydrophilic residues such as r, Thr, Asp or His are preferred. In particular, T hr and Set are preferred.

B can be one or more hydrophilic or hydrophobic amino acids, or a shorter peptide. A covalent bond may be used in the case of titanium. The difference in B is not critical, and water solubility and Desirable physical properties imparted to the entire peptide, such as resistance to xopeptidases Residues can be selected depending on their chemical and biochemical properties. The selection of D-amino acids is often When a D-amino acid is present at the end of a peptide, it increases resistance to exopeptidases. imparts resistance. When B is a covalent bond, it is advantageous for Q to be a D-amino acid. be. If B is not a covalent bond, B is preferably AIa, β-Ala, G ly, I Ie, Va l, Leu, Met, His, Lys, Arg, Glx , Asx, Cys, Ser or Thr or 1 or 2 D- or It is an L-amino acid.

To produce the smallest peptides of the invention that bind retroviral peptidases , B is preferably a covalent bond.

Y can be one or more amino acids or a covalent bond. one or more amino In the case of acids, they may be hydrophilic or hydrophobic. 1-3 residues are preferred, but longer It's okay. As in B, the residues in Y have the desired biochemical and Can be used to give physicochemical properties. This makes the use of hydrophobic residues less desirable. The carboxy-terminal D-amino acid provides exopeptidase solubility properties. It can be used to impart resistance to Preferred Y is Ala, Gly, IIe , Leu, Met, Va I, Arg, Lys, Thr.

1 to 3 amino acids selected from Ser, Cys, Glx, or Asx.

For smaller peptides of the invention, Y may be a covalent bond, but may include a single amino acid. Particularly appropriate. Ala, Gly, IIe, Met, Arg, Asx and Va t is preferred. Particularly preferred is valine.

Peptides in which M is Cha or Phe (4' R,) and d is 1 are used as substrates. peptides and are hydrolyzed into smaller peptides. Substrates are generally of any length. However, those with 6 to 9 residues are preferable. These are protease activity In addition, they can compete with natural viral substrates and thereby inhibiting viral replication and inhibiting disease progression in vivo. harm

However, the duration of action may be short due to metabolic instability.

Typically, these peptides are treated with a protease in a suitable buffered medium. It can be used to assay protease activity by reacting with Analysis of activity This can be done by detecting the hydrolytic cleavage of peptides. One such method is raw consisting of at least one separation of a synthetic peptide or its derivative and its quantification. It will be done.

In such assays, silica gel, Octadene 7 run, Sephadex, Chromatography using conventional solid supports such as ion exchange or adsorption resins One means can be used to separate the peptide products. Detection/quantification of products is performed using ultraviolet absorption. This can be done by extraction or other spectroscopic analysis. Alternatively, in one or more amino acids Incorporating radioisotopes such as tritium or 13C labels into the peptides , detection/quantification can be performed by radioactivity by substitution. As a further alternative, Fluorescence can be achieved by combining fluorescent residues such as the dansyl group at the amino terminus of peptides. Light detection/quantification can be performed. In addition to the fluorescent marker, the carboxy-terminal 3-(4-N - Quenching peptides, such as methylpyridyl)propyl oxene esters Fluorogenic methods that incorporate agents can also reduce protease activity. Useful for testing. Such methods are described, for example, by Dunn et al., Ana 1. Biochem, 129, 502 (1983), and the association of activity Provides continuous monitoring. These fluorogeninoc methods are used to produce hydrolyzed products. It has the advantage that hydrolysis can be directly observed without the need for separation.

Protease conjugation of M, Cha or (4'R,)PhaO peptides of the present invention Synthetic activity is indicated by its ability to act as a substrate. These peptides on the table of The substrate dynamics of

Table 1 Peptide binding to rHIV protease xtransmote a compound Re1.　 Km Kcat Example No. Vo Vo (m M) (m i n-’) 1 1.0 6.7423 9 1.0 6.9 4140 . 35 8.6 1641 neg,” --42 Yu, 34 1.9 2143 . 48 20 3044 . 4B 2.2.545 　　　　　　． 46 11’ 2146 1.23 2.3 1247 　　　　　、．． 36　　　　　　　B　　　　　　1548　 1.0 6゜8 3549 ．． 65 6.0 1B, 450 ．． 32 5.0 1.854 ．． 39, 71, 2.0355 . 86 6.5 23.056 1. 45 1.2 3157 neg, ”　　　　　　--59,563,38,0 60,548,01B 61　　　　　　　. 51 2.3 6.4 62, O5--63,0454,91,8 641,24-- 65,111914 661,2511,7183 95neg, ★ 12.8 5.396 　　　． 34 17.7 90.49B , 16 , 25 , 6599 　　　　． 09 6.2 1.4100 　　　　　　. 045 1.8 . 3810 1. 06 6.2 7.8 102 1.27 7.7 9611 0 1.29 9.8 103111 1.33 34 374115 　　　　． 48 7.1 16118 　’　neg, ★　　　　　　-119neg,”　　　　　　30　　　　 -121 ． 80, 96 16.6 Note・*neg ignored − No In other respects, those substrate peptides where M is Cha or Phe (4°R,) Indicator for finding amino acid sequences that bind to viral proteases It is useful as a protein, thereby inhibiting proteolytic activity. virus protect Modify such peptides that bind to enzymes to prevent hydrolysis of the peptides. This provides a means of prolonging the inhibition of protease activity.

Therefore, the group of compounds of the present invention includes peptides in which M is -NHCHR,R1-. is included. These peptides contain unnatural amino acids that are not α-amino acids.

These peptides do not have similar peptide bonds as the substrate, so they are naturally binds proteases in a manner that mimics the binding of natural substrates, but resists hydrolysis . Peptides of 2-9 residues are suitable. Peptides 1' of 3-6 residues are preferred. Better.

stomach. Preferably, since the unnatural amino acid can act like a 7-peptide, M is -N) iCHR,R,-1d is O and W is absent. In preferred L embodiments, R6 teeth In the formula, R', R1, R3', m and 0 have the same meaning as the formula ([) .

In a preferred embodiment, R8 is CHtCsH4R-, and R7 is CHt (01-t) CH, CI-(, CO-. Suitably, the peptide of formula (1) is -A l a-N [(CH (CH, Ph) CH (OH) CH, CH.

It has a partial structure of C0-Va 1-Va l-.

If infected or potentially infected animals are infected with the virus, It exists in the protease that viruses feed for processing polyproteins, but Luz's? J! ! R is inhibited and disease progression is prevented. Various retrowills As long as the presence of the Funcensus sequence is known in the polyprotein of the It can be broadly active against these retroviruses. cormorant like hepatitis virus The DNA virus, which depends on the protease contained in the virus, requires such treatment C. It can be a thousand sensitivities.

The following compounds are included, but the present invention is not limited thereto.

2-(acetylucerylglutaminyl asparaginyl)amino-3-phenylp Lopyl-prolylvalyl valinamide 2-(serylglutaminyl asparagini ) Amino-3-phenylpropyl-prolylvalyl Valinamide 2-(acetic acid) tylucerylglutaminyl asparaginyl)amino-3-hydroquine-1-oxy So-5-phenylpentylprolylvalyl valineamide 2-(acetylucerylglutaminyl asparaginyl)amino-3-hydroxy -1-oxo-5-phenylpentylvalyl valineamide 2-(serylglutaminylasparaginyl)ami/-3(S)-hydroquine-1 -oxo-5-phenylpentylvalyl 1 (phosphorus amide 2-((2-t-butoxycarbonyl-serylalanylalanyl)ami/-1- oxo-3-phenylpropyl)cyclopentylcarbonylvalyl valinemethy luester 2-(2-(,serylalanylalanyl)amino-1-oxo-3-phenyl Propyl) cyclopentyl-carbonylvalyl/Zarin methyl ester tri Fluoroacetate 2-(2-(t-butoxycarbonyl-serylalanylalanyl)amino-1 -Hydroxy-3-phenylpropyl)cyclopentylcarbonylvalyl Bali methyl ester 2-(1-hydroxy 3-phenyl-2-(serylalanyl) alanyl) aminopropyl) cyclopentylcarbonyl valyl valine methyl ester Stell trifluoroacetate 2-((],-]methquin-1-2-phenyl-1-(t-butoquinocarbonyl -serylalanylalanyl)amine)ethyl)phosphinyl)cyclopentylka Rubonylvalyl valine methyl ester 2-((1-hydroquine-1-(2-ph) phenyl-1-(serylalanylalanyl)amine)ethyl)phosphinyl)ink Lobentyl carbonyl-valyl valine methyl ester 5-(t-butoxycarboxylic) (bonyl-serylalanylalanyl)amino-4-hydroxy-1-oxo6- Phenylhexylvalylvaline methyl ester 5-(t-butoxycarbonyl-serylalanylalanyl)amino-4-hydro Quin-1-o-di-6-(4-hydrocydiphenyl)hexyl-valyl valine Methyl ester 4-(t-butoxycarbonyl-serylalanylalanyl)ami Nor-2,2-difluoro-3-hydroxy-1-oxo-5-phenylpentyl -Baryl valine methyl ester hydrochloride 4-(serylalanylalanyl)amino -2,2-difluoro-3-hydroxy-1-oxo-5-phenylpentylba Lil valine methyl ester hydrochloride 4-(serylalanylalanyl)amino-2,2-difluoro-1,3-dioki So-5-phenylpentyl-valyl valine methyl ester hydrochloride 5-(t-butoxycarbonylserylalanylalanyl)amino-4-hydroxy 7-6-(4-hydroxy)phenyl-1-oxo-hexineruvalylamide 5-((carbobenzyloxy-D-seryl)alanylalanyl)amino-4- Hydroxy-1-oxo-6-phenylpentylvalyl valine methyl ester 5-((D-seryl)alanylalanyl)amino-4-hydroxy-1-oxo -6-phenylpentylvalyl valine methyl ester 5-(t-butoxycarbonylserylalanylalanyl)amino-6-(4-hyperoxyalanyl) droxy)phenyl-4-hydroxy-1-oxo-hexylbaryl valine 5-(serylalanylalanyl)amino-6-(4-hydroxy)phenyl-4 -Hydroxy-1-oxo-hexylvalyl valine hydrochloride 2-((1-hydroxy-2-(serylalanylalanyl)amino-3-cyclo hexyl)-furovir)cyclobencunecarbonyl-valyl valine methyl es ter hydrochloride 2-((2-(t-butoxycarbonylserylalanylalanyl)a Mino-1-oxo-3-phenylpropyl)cyclopentylcarbonylvalyl Valinamide 2-(2-(serylalanylalanyl)amino-1-oxy-3-phenylp cyclopentylcarbonylvalylvalinamide trifluoroacetate 2-(2-<t--i toxycarbonylcerylalanylalanyl)amino- 1-Hydroxy-3-phenylpropyl)cyclopentylcarbonylvalyl phosphorus amide 2-(1-hydroxy-3-phenyl-2-(serylalanylalanyl)ami nopropyl) cyclopentylcarbonyl valyl valinamide trifluoroacetic acid salt 2-(, (1-methoxy-1-(2-phenyl-1-(t-butoxycarbonyl) -cerylalanylalanyl)amino)ethyl)phosphinyl)nclopentyl 2-((1-hydroxy 1-(2-phenyl-1) -(serylalanylalanyl)amino)ethyl)phosphinyl)cyclopentyl Carbonyl-valyl valinamide 5-(t-butofoxdicarbonylcerylalanylalanyl)amino-4-hydro xy-1-oxo-6-phenylhexylvalyl valineamide 5-(i-butofoxdicarbonylcerylalanylalanyl)amino-4-hydro xy-1-oxo-6-(4-hydroxy-phenyl)hexyl-valyl amide 4-<t-butoxycarbonyl-serylalanylalanyl)amino-2,2-di Fluoro-3-hydroxy-1-oxo-5-phenylpentyl-baryl Amido sulfate 4-(serylalanylalanyl)ami/-2,2-difluoro- 3-Hydroxy-1-oxo-5-phenylpentyl-valyl valinamide salt acid salt 4-(serylalanylalanyl)amino-2+ 2-difluoro-1,3-dit 47-5-phenylpentylvalyl valinamide hydrochloride 5-(carbobenzyloxy-D-seryl)alanylalanyl)amino-4-hyron Droxy-1-oxo-6-phenylhexylvalyl valineamide 5-((D-seryl)alanylalanyl)amino-4-hydroquine-1-ox So-6-phenylhexylvalyl Valinamide 5-(t-butoxycarbonyl lucerylalanylalanyl)amino-6-(4-hydroxy)phenyl-4-hydroxy Droxy-1-oxo-hexyl-valyl valineamide 5-(serylalanylalanyl)amino-6-(4-hydroxy)phenyl-4 -Hydroxy-1-oxo-hexyl-valyl valinamide hydrochloride 2-((1-hydroxy-2-(serylalanylalanyl)amino-3-cyclo hexyl)propyl)cyclobencunecarbonyl-valyl valinamide hydrochloride (4S,5S)-5-(,alanylalanyl)amino-4-hydroboxy-1-o xo-6-(4-hydroquinphenyl)hexyl-valyl valine methyl ester acetate (4S,5S)-5-((carbobenzyloxy-β-alanyl)alanyl)a Mino-4-hydroxy-1-oxo6-(4-hydroxyphenyl)hexyl -Valyl valine methyl ester (4,S,5S)-5-(, (β-alanyl) alanyl)amino-4-hydroquine-1-oxo-6-(4-hydroxyphenylene) ) Hekinruvalyl valine methyl ester hydrochloride (4S, 5S)-5-(A ranylalanyl)amino-4-hydroquine-1-oxo-6-cyclohequinol Hekinrubalyl valine methyl ester (4S,5S)-5-(carbobenzyloxyalanylalanyl)amino-4- Hydroxy-1-oxo-6-phenylhexylvalyl valine methyl ester le (4S,5S)-5-(alanylalanyl)amino-4-hydroxy-1-oxy So-6-phenylhexyneruvalyl valine methyl ester (4S,5S)-5-(alanylalanyl)amino-4-human0x-1-ox So-6-(4-hydroquinphenyl)hequinrubalyl Valinamide acetate (4S,5S)-5-((carbobenzyloxy-β-alanyl)alanyl)a Mino-4-hydroquine-1-oxo-6-(4-hydroxyphenyl)hexyl -Baryl (4S,5S)-5-((β-alanyl)alanyl)amino-4-hydro xy-1-oxo-6-(4-hydroxyphenyl)hexyl-valyl valyl amide hydrochloride (4S.5S)-5-(alanylalanyl)amino-4-hydroquin-1-o Xo-6-enchlorohexyl-hequinrubalyl valineamide (4S,5S)-5-(carbobenzyloxyalanylalanyl)amino-4 -Hydroxy-1-oxo-6-phenylhequinrubalyl Valinamide (4S,5S)-5-(alanylalanyl)amino-4-hydroquine-1-ox So-6-phenylhekifluvalyl valineamide (4S,5S)-5~(cal bobenzyloxyalanyl)amino-4-hydroxy-1-oxo-6-phenylated Ruhekifluvalylvaline methyl ester (4S,5S)-5-(alanyl)amine-4-hydroquine-1-oxo-6 -Phenylhexyl-valyl valine acetate (4S,5S)-5-(t-buto oxycarbonylalanyl)amino-4-hydroquine-1-oxo-6-(4- hydroxyphenyl)hexyl-valyl valine methyl ester (4S, 5S) -5-alanylamino-4-hydroxy-1-oxo-6-(4-hydroxy phenyl)hequinruvalyl valine methyl ester acetate (4S,5S)-5-(,carbobenzyloxyalanyl)amino-4-hydroxy Doroki/-1-oxo-6-phenylhequinruvalyl valineamide (4S,5S)-5-(alanyl)amino-4-hydroxy-1-oxo-6- Phenylhequinruvalyl Valinamide (4S, 5S)-5-(t-butoxy carbonyl)alanyl)amino-4-hydroquine-I-oxy-6-(4-hydro roxyphenyl) hequinrubalyl valineamide (4S,5S)-5-(alanyl)amino-4-hydroxy-1-oxo-6- (4-Hydroquinphenyl)hexyl-valyl valinamide acetate (4S,5S)-5-amino-6-(4-hydroquine)phenyl-4-hydroxy N-1-oxohequinrubalyl valine benzyl ester hydrochloride (IR,2R)-2-(((JS,2S)-1-hydroxy-2-amino-3- 7chlorohexyl)propyl)-cyclopenta7carbonylrubaryl valinemethy ester hydrochloride (4S,5S)-5-amino-4-hydroquine-1-oxo- 6-Phenylhexylvalyl valine methyl ester hydrochloride (4S, 5S)- 5-(t-butquinecarbonyl)aceno 4-hydroxy-1-oxo-6-ph enylhequinruvalyl valine methyl ester (4S,5S)-5-amino-6-(4-hydroxy)phenyl-4-hydroxy C-1-oxo-hexyl-valyl valinamide hydrochloride (IR,2R)-2-(((is,2S)-1-hydroxy-2-amino-3- cyclohexyl)propyl)-cyclopentanecarbonyl-valyl valinami Hydrochloride (4S,5S)-5-amino-4-hydroxy-1-oxo-6-phenylhexyl Diruvalyl Valinamide Hydrochloride (4S, 5S)-5-(t-butoxycarboxylate) amino-4-hydroquine-1-oxo-6-phenylhequinrubaryl Valinamide (5S)-5-(carbobenzyloxyalanyl)amino-4-hydroxy-1 -Oxo-6-phenylhexylvaline isobutyramide (5S)-5-(myristyl)amino-4-hydroquine-1-oxo-6-phene Nylhexyl valine isobutyramide (5S)-5-(, carbobenzylo xyalanylalanyl) to amino-4-hydroxy-1-oxo-6-phenyl xyl varifuxo 6-phenylhexyl valine isobutyramide 5-( carbobenzyloxyalanyl)amino-4-hydroquine-2-methyl-1-o Xo-6-phenylhexylvalyl valine methyl ester 7-Methyl-5-(carbobenzyloxyalanyl)amino-3°4-dihydro xy-2-phenyl-I・-oxo-octylvaline isobutyramide 5-(carbobenzyloxyalanylalanyl)amino-4-hydroxy-1- Oxo-6-phenylhexylvaline isobutyramide 5-(carbobenzyloxyalanylalanyl)ami/-6-phenyl-4-hyron Droxy-(1-oxo)hexyl-valyl valinecarbobenzyloxyara Nyl-(3-hydroxy-5-amino-6-phenyl)hexanoylbasol (4-aminomethyl)pyridine 5-(carbobenzyloxyalanylalanyl)amino-4-hydroxy-6- Phenyl(1-oxo)hexyl-valine isobutyramide (4S,5S)-5-(methoxycarbonylalanylalanyl)amino-6-ph phenyl-4-hydroxy-1-oxo 1.7 (4S,5S)-5-(carbobenzyloxyalanylalanyl)amino-6- Phenyl-4-hydroquine-1-oxo-hequineruvalyl valine (4S,5S)-5-(alanylalanyl)amino-6-phenyl-4-hydro Quin-1-oxo-hexyl-valyl valine (4R3,5S)-5-((t- butquincarbonyl)isoleucyl)amino-7-methyl-4-hydroxy-1 -oxo-octyl-loy/ruu (○-benzylene aspartate methyl ester) and (4R3,5S)-5-((t-butoxycarbonyl)inloynol)amino- 7-Methyl-4-hydroxy-1-oxo-octyl-loinruasparagine Acid methyl ester.

Certain individually preferred compounds are as follows.

(4S,5S)-5-(carbobenneluoquine-γranylalanyl)amino- 6-phenyl-4-hydroquine-1-oxohexylbaryl valine methyl es Tell (4S, 5S) 5 (alanylalanyl)amino-6-phenyl-4-hyron Doroquin-1-oxo-hexylvalyl valine methyl ester (4S,5S)-5-(carbobenzyloquinalanyl)amino-6-phenyl -4-Hydroquine-1-oxo-hexylhalylvaline methyl ester (4S,5S)-5-(alanyl)amino-6-phenyl-4-hydroxy-1 -oxo-hexylvalyl valine methyl ester and (4R,5S-5-((t-butoxycarbonyl)isoleucyl)amino-7 -Methyl-4-hydroquine-1-oxo-octyl-leucyl-(0-benzyl ) aspartic acid methyl ester and (these carbonyl/acid and carboxylic acids) Quinamide.

The peptide of the present invention is prepared by coupling appropriate amino acid residues and, if desired, protecting the peptide. 2 and optionally modify the amino or carboxyl/terminus of the peptide. Manufactured by. These are preferably based on Merryfield's solid phase technology (J ,Am,Chem,Soc,.

85.2149 (1964)), but known solution methods can also be used. . Convergence of solution method or combination of solid phase method and solution method (c.

di-, tri-, tetra- or pentape The fragments are prepared by solid-phase or solution methods7, and other G-, T-, or is coupled with a tetra-peptide and further modified using a solution method. [Solid phase method Peptide synthesis (Solid Phase Peptide 5ynthes is) J or [Peptide synthesis (Peptide 5 synthesis) J] Methods disclosed in the literature can be used to produce the peptides of the invention.

Each amino acid or peptide is suitably protected by methods known in the peptide art. . Boc or carbohensyloxy group retains an amino group, especially the amine 7 group at the α position. favorable for protection. A benzyl group or an appropriately substituted benzyl group is a cysteine group. mercapto group or serine or threo group of thiol-containing amino acids Used to protect the hydroxyl group of nin. Tosyl or nitro is guani of Arg For the protection of the imidazole of Zine or His, an appropriately substituted carbobenzyloyl The xy group or benzyl group is the hydroxy or link group of Tyr, Ser or Thr. It can be used to protect the ε-amino group of gin.

Suitably substituted carbobenzyloquine or benzyl groups include chloro, bromo, dichloro, Ortho- and/or para-substituted with tho- or methyl, and the opposite of the protecting group. Used to modify responsiveness. Cysteine and other sulfur-containing amino acids are thioalkyl or can also be protected by forming a disulfide with a thioaryl. Boc group Most conveniently, the protecting groups are those that are not removed by mild acid treatment.

These protecting groups can be prepared by known catalytic hydrogenation, sodium or H in liquid ammonia. It is removed by F treatment.

When using solid-phase methods, start at the carboxy terminus and move toward the amine terminus of the peptide. The peptides are sequentially constructed. Solid phase synthesis or C-terminus of protected amino acid with benzene Drylamine resin (BHA), Methyl hexahydrylamine resin (MBHA) or chloromethyl resin (CMR), as described in U.S. Pat. The covalent bonding is initiated as described in Section 6. The carboxy terminus of the generated peptide In the case of carboxamides, BHA or MBHA carrier resins are used. CMR is generally used for the carboxy terminus or carboxy group of the resulting peptide. , which can also be used for the production of carphokidiamides or esters.

Once the first protected amino acid (AA) has been coupled to the desired resin, a mild acid treatment The amine 7 group is hydrolyzed by the process, and the free sulfoquine group of the second protected AA is converted to this amine. Coupled to 7 groups. Formation of desired peptides without isolation of intermediates Repeat this operation one after another until the The generated peptides can be added in any order. However, deprotection and/or separation from the resin can be performed.

Treatment of CMR-loaded peptides with HF or HBr/acetic acid removes the peptide from the resin. and yields the carboxy-terminal amino acid in the form of a carboxylic acid. CMR person Treatment of peptides with ammonia or alkyl amines in alcoholic solvents produces carboxinamide or alkylcarboxinamide at the carboxy terminus. Cheating.

If an ester is desired, the CMR resin is diluted with methyl, ethyl amine in the presence of triethylamine. treated with a suitable alcohol such as alcohol, propyl, butyl or benzyl alcohol. The peptide is separated from the resin and the ester is produced directly.

The ester of the peptide of the present invention can be produced from a carboxylic acid precursor by a conventional method. You can also do it. Typically, a carboxylic acid is treated with an alcohol in the presence of an acid catalyst.

Alternatively, the carboxylic acid can be prepared as an acid compound, activated anhydride or ester. to the active acyl intermediate, preferably by treatment with an alcohol in the presence of a base. can be done.

A preferred method of removing the peptide from the carrier resin is using anisole or dimethquinone. Dry the resin-supported peptide in the presence of a suitable cation scavenger such as It is treated with dry HF. This method, except for the thioalkyl group-protecting sulfur, All protecting groups are removed and the peptide is released from the resin at the same time. In this way, The peptide hydrolyzed from CMR is a carboxylic acid, and the peptide removed from BHA Tido is obtained as carboxamide.

As is known, alkylation or acetylation is used to modify the terminal amine groups of peptides. file format is carried out. These modifications can be made before combining the amino acid with the peptide. Alternatively, after the peptide has been synthesized and the terminal amine 7 group has been generated, but the protecting group This can be done before the removal of

Typically, acetylation involves converting the corresponding alkyl acid into an acylate in the presence of a tertiary amine. The reaction is carried out on free amide 7 groups using halides, anhydrides or active esters.

Monoalkylation is possible with lithium or sodium cyanoborohydride. in the presence of a mild reducing agent, suitably with an aliphatic aldehyde or ketone. This is conveniently carried out by reductive alkylation of alkylation converts the amine group into a base. This can be carried out by treatment with an excess of alkyl halide in the presence of alkyl halide. The long chain atom of the present invention Alkyl and aral groups are considered advantageous due to their affinity for lipid membranes. example For example, myristylation and stearylation are useful.

Solution synthesis of peptides can be carried out by conventional methods that generate amide bonds.

Typically, a Boc-amino acid with a free carboxyl group is optionally added to 1 - Hydroquine benzotriazole (HOBT) and dimethylaminopyridine (DM N,N'-dicyclohexylcarbodiimide in the presence of a catalyst such as AP'l Using a suitable carbodiimide coupling agent such as (DCC), free amine Coupling with a protected amino acid having a group. Free capacity of protected Boc-amino acids Formation of active esters, anhydrides or acid halides of the carboxyl group, then as desired Other methods, such as the reaction of a protected amino acid with a free amino group in the presence of a base, The law is also appropriate. For example, Hoi! Boc-amino acid or peptide Methylene chloride in the presence of a base such as lumorpholine, DMAP or a tertiary amine In a dry solvent such as THF, treatment with inbutyl chloroformate 1, which is further combined with a second protected amino acid or the free atom of the peptide. React with Mi7 group. The peptide obtained by this method can be extracted with its amine by a conventional method. Alternatively, selectively remove (&MJ) with carboxamide, and use the same method to Coupling with peptides or amino acids.

Once the peptide is completed, it is hydrogenated in the presence of a platinum or palladium catalyst as described above. treatment with sodium, chloride, hydrohydric acid or alkali in liquid ammonia. Protecting groups can be removed.

In solution synthesis, esters are often used to protect the terminal carboxyl group of peptides. used. These are potassium hydroxide or sodium carbonate in hydroalcoholic solutions. Treatment with alkali metal hydroxides or carbonates such as thorium to form carboxylic acids It can be changed. As mentioned above, the acid passes through the active acyl intermediate and becomes the ester. It can be changed.

Amides and substituted amides of the invention can be similarly prepared from peptide carboxylic acids. Wear. i.e., by reacting ammonia or a substituted amide with an activated acyl intermediate. Amide can be obtained. The use of coupling agents such as DCC It is convenient for the formation of substituted amides from itself and the appropriate amine.

In addition, the methyl esters of the present invention can be used in methanol solution with ammonia or substituted Can be converted directly to amides or substituted amides by treatment with amines. Ru. A methanol solution of peptide methyl ester is saturated with ammonia and pressurized. Stir in the reactor to obtain the simple carboxamide of the peptide. Carbokinami Do is more stable than ester and is preferred.

The amino acids constituting the M residue of the peptide are phenylalanine, its derivatives, or - Contains unnatural amino acids designated as NHCHR,R,-.

Derivatives of phenylalanine include tyrosine, (O-C,, alkyl)tyrosine, 4'-halophenylalanine, 4'-nitrophenylalanine and 4'-a Includes minophenylalanine.

Intermediates useful in the synthesis of peptides of the invention, where M is -NHCHR,R, have the formula (lea ): (eea) is shown.

In the formula, R, is independently C,, A l k, (CH, n S C + -s A l k, (CH+) n OC+-sA l k, (CHt) ncs-7 sik0f Lu + Lu or (CHz) m CaHa Ra: Ra is halogen, OR', Not, NH, tri, Ii! -1; R1 is (CH2), -, CI(R3(CH21,Co+, Co(CH2)rlICo +.

CHR3CHR31(C821mCo-, CHR3CHR31CHR4(CH2 ) IT, Co-.

CHR3CHRICHR' (C) +211TICo +, CHR3CHR' CHR1(CH2)mCo-.

C0CHRI CHR’ I CH2) mco-C0CF 2CR’ R” (CH21mco-t C0CF2 (CH2) mco-■ CH"C0CHRI(C)!2 Co+, C0CH-CR'(CH2)mC o-.

CH25(0)pCHR’(C)! 2) ITICo-, CH2NR’CHR” ( CH2 deception CO-.

P■O(OR') (CH2)mCo-.

R3 and Rlo are each independently OH, H or N H1.

R is OH, H, NH, or =0.

m is independently O, l, 2 or 3.

n is independently l or 2.

p is 0, l or 2.

R1 is H.

R8 is CH3Co, C, -sA Ik, Dns, Cbz or Boc.

Alternatively, R and R1 together form phthalimide.

X is H1 halogen, OR", OC,, Δlk, NHR' Ro or 0COC ,, AIk.

Ro is H or C, Alk.

R'' is H or C1-, , AIk.

These unnatural amino acids can themselves be used as intermediates for the production of protease inhibitors. It has the usefulness of Therefore, the peptide substrate that binds to the protease, One or both of the amino acids are replaced at the cleavage site with a suitable compound of formula (Ila). It can be converted into an inhibitor by converting it into an inhibitor.

Unnatural amino acids are generally not alpha-amino acids, but can be derived from alpha-amino acids. Ru. That is, the identity of R1 is established by the selection of amino acids of formula (II1). It will be done.

In formula (II), R is selected as described above.

Boc, modification of the amine 7 group of α-amino acids, such as by acetyl or phthaloyl groups. protection and acid or ester to alcohol of formula (IV) or to alcohol of formula (■) Conversion to dehyde is similar to the preparation of the compound of formula (I[a) of the present invention for introduction of R3 substituent. Provides a flexible intermediate for N-methoxy-methylene of these α-amino acids Ruamides are also useful intermediates for this application. α-Amino acids can be obtained from natural sources. It can also be synthesized by many methods well known in the peptide field.

Alcohols of formula (IV) can be obtained by Chevrolet reduction of the corresponding protected amino acids. can. The aldehyde of formula (V) is a methyl ester of the corresponding amino acid, e.g. , can be obtained by reduction with diisobutylaluminum hydride (DIBAL) . Alternatively, the alcohol of formula (IV) can be used as a Dess-Martin biogenane. A mild oxidizing agent such as can be oxidized to aldehydes using amines).

For example, if R1 is Alk, alanine, valine, leucine and isoleucine is particularly useful for producing xyl aldehydes. α-aminobutyric acid, α-aminobutyric acid Minoisobutyric acid and α-aminopentanoic acid are commercially available as racemates. Ru. In general, any alkyl-α-amino acid can be used such as triethylamine, DBU or with a suitable alkylnolide in the presence of a base such as or other tertiary amine. It can be synthesized by alkylating nitroacetic acid. Sodium metkind also Alkoxides such as ethoxide are also suitable bases. α-nitroalkyl acid Alternatively, esters can be reduced with hydrogen and palladium or platinum catalysts to produce amino acids. Obtain noic acid. Alternatively, the methyl or ethyl ester of N-benzylidene Alkylation of esters such as esters is also common in the production of α-amino acids. .

That is, the henglycine is treated with sodium or potassium or lithium hydride. Treatment with a strong base such as diisopropylamide, followed by a suitable alkyl chloride The desired amino acid is obtained by alkylation with can get. These methods include, for example, /chloroalkylmethylbromide or 2- By using cycloalkylethyl bromide, cycloalkylamino acids It can also be used for manufacturing. Another method for xyl amino acid synthesis is Tet.

Let, 1123 (1987).

Most amino acids where R is aryl or aralkyl are commercially available; Phenylalanine, 4゛　-halo and 47-nito-0phenylalanine, tyrosine (○-alkyl)thironne, phenylglycine and (4゛-hydroxy) Like phenylglycine (what is known). Homologues of these compounds The compound can be produced by conventional methods of amino acid synthesis. i.e. nitroacetate or N-henjilidene green as 1-phenyl-2-bromoethane or 1- Phenyl-3-bromopropane was alkylated as described above to give the homologous amino acid. Ru. α-Amino acids containing oxygen or sulfur in R1 include serine, homoserine, threo derived from nin, cysteine or methionine, or other than methionine obtained by alkylation of amino acids. Alkylation is the corresponding thiol compound bases such as triethylamine, DBU or sodium hydride, and a suitable alkyl such as thyl, ethyl, isopropyl or inbutyl bromide This can be done by treatment with halide.

The di-homologous compound can be prepared by the conventional method for producing amino acids as described above.

Therefore, esters of nitroacetic acid can be mixed with bases and acrolein as described above. Process with frog reaction. The obtained aldehyde was then dissolved in sodium borohydride. The mild compound is reduced with a reducing agent to obtain the desired hydroquine group. Then, a nitro group is attached. Catalytic reduction gives α-amino 5-hydroxypencunic acid. Corresponding mercapto class Analogs can be synthesized from the pidrox/intermediate. i.e. amino and carboxy After protecting the hydroxy group by known methods, the hydroxy group is protected with triethylamine or other Treated with phosphorus tribromide or oxalyl bromide or chloride in the presence of a tertiary amine , converted to halide by treatment with triphenylphosphine and carbon tetrabromide. I can do it. This halide is treated with sodium sulfide or potassium thioacetate and then Saponification with sodium hydroxide yields 5-mercap 1--2-aminopentanoic acid. .

This intermediate can be further alkylated in the same manner as for nstine or serine. .

As mentioned above, α-amino acids are aldehydes or alkaline acids of formulas (iv) and (V). Can be turned into a call. Then, substituent R1 is introduced into these intermediates. By this, a compound of formula (ITa) of the present invention is obtained.

Introduction of a hydrocarbon substituent to R6 can be achieved by converting the aldehyde of formula (V) into a Grignard reagent. This can be carried out suitably by reacting with organometallic hydrocarbons or organolithium species. . The initial carboxylic acid functionality of R2 can be protected in the form of an orthoester or Disguise yourself as Finn.

-T(D) In specific examples, the organometallic species is lithium or magnesium and allyl bromide, 1-bromo-3-butene, 1-bromo-4-pentene or 1 -Magnesium produced from alkylene halides such as bromo-5-hexene or lithium alkylide, or an alkyl-substituted derivative thereof. Ru.

When an organometallic reagent is added to the aldehyde of formula (V), the formula (Via) or (VI The hydroquine alkene b) is obtained. Here, Roi;! OH,R,’ is H,C,5A　Ik,(CH,)　LloC,-,A,l　k.

(CH,) nC3-77 chloroalkyl or (cHt)1mceH4-Ra , and other symbols are the same as in formula (lla).

(VIa) (VIIal(VIb) (From VI) Alternatively, the Gutsnia test Drug-like organometallic alkylidene formula (■rl) amino acid N-methoxymethylidene By adding amide, the formulas (VIa) and (VIb) with Ro = 0 are Obtain Toalkene. 1-metallo-2-propenylcyclopentane or 1-metallo-2-propenylcyclopentane The corresponding formula (■ [c) (where R1, R6, R,, R', R'', m and n are of the formula (IIa ) to obtain the same 7-chloroalkene. Therefore, R, is HlC,,Alk, ( CH,)QC,-, A Ik, (CH,)C3-, cyclofurkyl or (C H2), C, H, -Ra, formula (VIIa) where other symbols are the same as formula (IIa), The method for producing the compound of (Vllb) or (VIIC) is based on the formula (VIa), ( VIb) or (VIc) with the formula (VI Ia) where R4 is ;O and X is OH , (VIIb) or (Vllc), and then optionally then reduce or reductively ainate the keto group in either order, optionally adding carboxylic acid. converting phosphoric acids into esters, amides, aldehydes, anhydrides or acyl halides. It consists of Suitably, Ro is HSm is O, preferably a compound of formula (Vlla) In this case, R4' is H.

Hydroxy/or ketoalkane oxidation can be accomplished by conventional methods such as with potassium permanganate. or osinolysis followed by standard dimethyl sulfide treatment and further can be done by treatment with potassium permanganate or Jones reagent. Ru. As is clear, this method is not applicable when R. contains a sulfur substituent.

Other methods applicable to this type of compounds are J. Med., Chem., 30°3 74-83 (1,987) and J, ○rg, Chem,, 51°21.39 21. -26 (1986).

Formula (VIIa) using a common reducing agent such as sodium borohydride, ( Vllb) and (VIIc) (Ro is -0) for the reduction of keto compounds The corresponding alcohol is obtained. If alcohol is desired, benzyl or protection by the formation of silyl ethers or acetates, such as lactone formation. To prevent undesirable side reactions and improve subsequent incorporation into the peptides of the present invention. is desirable.

Protection of such alcohols can occur before or after derivatization into carboxylic acids. . Hydroxy7 can be further treated with halogenating reagents such as phosphorus trichloride or phosphorus tribromide. can be converted to chloride or bromide, which can then be converted to hydrogen or Raneynicke. can be reduced to a methylene group with If an amine 7 group is desired, the keto group is Ammoniums such as anoborohylolate and ammonium chloride or bromide Can be reductively aminated with halides. Carboxylic acids can be converted into esters and amino acids using known methods. It can be converted into anhydride, alderide, anhydride or norodane.

Introducing unsaturation at R7 is carried out from the amino acid product corresponding to formula (Vll). I can. Reduction of the carbonyl group to a hydroquine group as described above, followed by hydrogenation. Removal of the doloquine group yields the unsaturated compounds of the invention. The hydroxyl group is DBU or lithium diisopropylene instead of menlate or halide, as in This can be done by treatment with a base such as pyramide. Mild mineral acid (HC Other methods of hydroxy group removal such as by 1, H, SO2) are also suitable. R3 Other methods of introducing unsaturation into the Witsch olefins of aldehydes of formula (V) include is included. Hydrogen and palladium catalyzed reduction of these unsaturated compounds presents a comprehensive alternative method for the removal of the ketone group of compounds of formula (VII) (R, = 0). provide

Introduction of the nitrogen substituent at R2 is carried out by converting the amino acid of formula (V) into Cl-4 under reducing conditions. Aminoalkanoic acid, alkylamino CI4alkanoic acid or amino7 moiety is saturated or Alternatively, it can be carried out by reacting with a structure containing an unsaturated 5- or 6-membered ring. The ami Examples include glycine, alanine, 3-aminopentanoic acid, 4-aminopentanoic acid. , phthalic acid, proline, Δ3-tehydrobroline and 2-calhoki/piperazi and methyl esters. Under suitable conditions, hydrogen and Application of radium or platinum catalyst or sodium cyanoholohydrate is included. Thus, the formula (V I I I) (where each symbol represents the formula (ila) The method for producing the compound represented by (which is the same as Lolin, Δ3-dehydrobrolino or 2-carboxypiperazine ester or The ester is reacted with a carboxylic acid or an acid, and then the ester is reacted with a carboxylic acid, an Convert acids to esters, aldehydes or amides or convert acids to esters, amides, aldehydes It consists of changing it to a do or a/al halide.

To introduce a sulfur substituent to R6, a thioalkanoic acid or ester is added to an α-amino acid. This can be done by reacting with a derivative.

Phosphorous tribromide, oxy salt in the presence of a base such as triethylamine or pyridine Hydroxyl of formula (IV) using conventional reagents like phosphorous chloride or thionyl chloride. By changing the amino acid to the corresponding halide, in the presence of a base, a thioalkanoic acid or an ether Process with stealth. The sulfide thus obtained was treated with sodium metaperiodate. treatment with mild oxidizing agents or sulfones, such as potassium permanganate. can be oxidized to sulfoxide by treatment with

The introduction of a fluorocarbon substituent at R2, where the fluorine is at γ of the amine 7 group, is represented by the formula ( V) by treating the aldehyde with ethyl difluorobromoacetate and an activated zinc layer. (see J. Med. Chem., 29°2080-87 (1986)).

Congeners of such compounds are alpha-keto diesters such as dimethyl oxaloacetate. It can be manufactured from.

For example, dimethyl oxaloacetate can be converted to diethylaminosulfur trifluoride (DAS). T) to obtain α-difluorodimethylsuccinate. The α-difluoro Difluoroalcohol by selectively reducing the ester group with sodium borohydrate 7s reagent, Dessmartin periodinane or swarmer solution. It can be oxidized to the corresponding aldehyde by mild oxidation methods such as the oxidation method. this Difluoroaldehyde is converted to 2-difluoroaldehyde in the presence of DBU or a base such as potassium carbonate. Formula R+ CHt such as trophenylethane or 1-nitro CI4 alkene Condensation with a nitro compound of N O2, followed by hydrogen and 5% Pd/C or platinum black catalyst. Reduction of the nitro group with a medium provides the desired difluoro congener. these Alkylated derivatives of homologs of are obtained by salting the initial α-keto diester prior to said treatment. It can be produced by alkylation with a group and an alkyl halide. For example, D Alkylate dimethyl oxalacetate with 2 equivalents of methyl iodide in the presence of BU and 2-keto-3,3-nomethylsuccinate are obtained. As mentioned above, Converted to 2,2-dimethyl-3,3-difluoro-4-oxo-methylbutanoate. When condensed with 2-phenylnitroethane and reduced, 2,2-dimethyl-3, 3-difluoro-4-hydroquine-5-amino-6-phenylmethylhexanoe You can get a discount. These compounds can also be further oxidized to fluoroketones. At this stage, it is preferable that the i & to do. Mofumasoto oxidation (DMSO, acetic anhydride), Dess Martin Periodina or pyridine/chromium trioxide are suitable oxidizing agents.

Introduction of phosphinyl into Rt was performed by converting benzyl carbamate to triphenyl in acetic acid. Phosphite and phenylacetic aldehyde, isobutyraldehyde or is condensed with an aldehyde of formula R, CHO such as 2-methylbutyraldehyde. Formula (IX): (Eng.X) This can be done by obtaining the phosphonate of

Further treatment of this phosphonate with an alkoxide yields the corresponding phosphonic acid monoamine. A rukyl ester is obtained. For example, treatment with sodium methoxide monomethyl ester of acid is obtained.

The phosphonic acid is treated with a suitable halogenating agent such as thionyl chloride or thionyl bromide. Reactive intermediate suitable for side chain modification by conversion to phosphonyl halide with is obtained. Lithium or alkyl butromide, fromo-3-butene, 2- (2-methylpropenyl)chlorochloropentane or 2-(2-methylpropenyl) Bell) - Cyclo with organometallic reagents such as Grignard reagents derived from bovine chloride. By processing formulas (Xa) and (Xb)': (Xal) (Xk)1 (where R″°° is C1, The alkyl methyl phosphite of the present invention has the same alkyl and pond symbols as in formula (Ila). F-1. is obtained. Then, as mentioned above, potassium permanganate or osino The corresponding alkyl phosphinate carboxylic acid of the present invention is obtained by oxidation by lysis. Ru.

Therefore, formula (XI) (X Engineering a)　　　　　　　　　　　　　　　(From The compound of formula (Ila)) is a compound of formula (Xa) or (xb) oxidizes to phosphinic acid and optionally hydrolyzes the alkyl phosphinate to phosphinic acid. and, if desired, convert the carboxylic acid into an ester, amide, aldehyde, anhydride or Produced by converting to sil halide. Alkyl phosphinate phosph Hydrolysis to phosphoric acid can be carried out by treatment with a strong acid such as hydrobromic acid. It's obvious In particular, this method is not applicable when R contains a sulfur substituent. These transformations Methods applicable to compounds are described, for example, in J.

Med, Chem, 30.1603-09 (1987).

A further aspect of the invention is that the peptide, when incorporated into the peptide, The present invention relates to the production of completely new amino acids that provide hydrolyzable imitations. this These amino acids were prepared as described above and are further illustrated in the Examples below. child These are incorporated into the peptide of the present invention by a method known per se, and are converted into the peptide of the present invention by a method known per se. the law of nature) It is indicated by.

In the formula, R4 is independently C,,Alk, (CH-)nSC1-sAl k, -(CHm)nOc, -, A I'K, (CHt)n C3-t ノ”　　　　7　Jl/　　　　　　/l/　*　　　　　　　-(CHt)mcaHa-Ra　 ; R, a is halogen, No, 0H1QC, -5A lk, NH, or Hl Rl is fCH21n-X, (CH2), Co-X, CHR3C1(R4CHR'(C )! 2>IT, Co+.

C0CRICHR’fC)! 21+nco-x, CHR3CF'2 (CH2) ,,Co-X, C0CF'2CR'R" (CH211T, bo-X.

CHR3(CH21mCo-X, Co(CH2)mCo-X.

R3 is H, NH, or OH.

1? , , itH, NH, , CHm or =O.

R6 is H.

Ro is CH3CO, C, -5Alk, Dns, Cbz or Boc or R6 and R6 together form phthalimide.

X is hydrogen, halogen, OH, ○C1-, Alk, NHR’R” or 0C OC1-, Alk.

R' and R'' are hydrogen or C,-8Alk.

m is independently 0.1.2 or 3.

n is independently 1 or 2.

However, R1 is phenylmethylene, isobutyl or cyclohex/lumethylene In the case, R3 is CHR, CH, Co-X, C0CH, C0-X, C0CF, Co- X* or CHR, CF, Co-X (when R1 is isobutyl, R9 is C Not H(OH)CH, CH, Co-X or C0CH, CH, Co-X.

1 or 2 that are subject to degradation by proteases, especially retroviral proteases Incorporation of a compound of formula (Tlb) into the peptide in place of the amino acid residue of Methods of producing protease inhibitory activity are provided. The above synthesis method is non-selective , or racemically introduce a xyl center. As is often a problem in biological systems , a single xyl-centered arrangement is usually optimal. In many compounds, one or more There is a xyl center in the compound, which changes the xyl center noastereomerism of the compound. this These diastereomers are usually analyzed by gel chromatography using a suitable mobile phase. It can be separated using a 7-run chromatography carrier or an octadecyl chromatography carrier. hmm When using silica gel, diastereomers can be eluted using ethyl acetate and beef sanitation. or a mixture of methanol and chloroform can be used. Octadecyl 7 rank If using a chromatographic carrier, use water and methanol or anidonitrile. Diastereomers are killed with Mu using a mixture of

For acidic or basic compounds, the mobile phase is 0.020.5% acetic acid, trifluorocarbon It can be buffered by the addition of acetate or phosphate buffers.

The residue denoted by M has only one chiral center or it is difficult to separate diastereomers. In difficult cases, xyl centers of other constituent amino acids may be incorporated into the peptide to provide further asymmetry. The resulting diastereomers can be separated in a later step. But long In some cases, it may be preferable to separate diastereomers; separation of the xyl center is not essential.

The unnatural amino acid M residue can be incorporated into a peptide using a known method as described above. I'll come.

After deprotection, acid addition salts of the final peptide can be prepared.

Acid addition salts of the peptides are prepared by adding hydrochloric acid, hydrobromic acid, or sulfuric acid to the peptide, preferably in a solvent. , phosphoric acid, acetic acid, maleic acid, succinic acid or methanesulfonic acid. It can be manufactured according to conventional methods. Acetate salts are particularly preferred. The final peptide has acidic groups. , a cationic salt can be produced. Typically, the compound is added in excess of a suitable With alkaline reagents such as hydroxides, carbonates or alkoxides containing cations Process. Cations present in pharmaceutically acceptable salts include sodium and potassium. Examples include ammonium, calcium and ammonium cations. Certain compounds are not allowed. Forms inert salts with the zwitterions it contains.

The compound of formula (1) where M is -HNCHR,R,- can be used to infect susceptible viruses and treat used to induce antiviral activity in patients in need. Treatment consists of an effective amount of The selected compound is preferably dispersed in a pharmaceutical carrier [2] for oral, parenteral, or buccal administration. , administered transdermally, rectally or by inhalation. Active ingredient dosage unit The amount is selected from the range of 0.05-15 mg/kg. For acute and chronic infections, These dosage units are administered 1-10 times per day. M is -HNCHR,R , - the protease inhibitory activity of the peptide of the present invention is about 10 nM - about 250 μM indicated by the ability to inhibit hydrolysis of peptide substrates by rHIV in the range of . child The inhibition constants of these peptides are shown in the table below.

Table 2 Inhibition of rHIV protease Compound Ki Example No. (μM) The pharmaceutical composition of the peptide of the present invention or its derivative Formulated as a solution or lyophilized powder for oral administration. The powder may be mixed with a suitable diluent or or other pharmaceutically acceptable carrier prior to use. Liquid formulations are generally buffered It is an isotonic aqueous solution. Examples of suitable diluents include isotonic saline, standard 5% Aqueous dextrose solution or buffered sodium or ammonium acetate solution is included. Included. Such formulations are particularly suitable for parenteral administration, but are also suitable for oral administration and inhalation. Can be used.

Polyvinylpyrrolidone, gelatin, hydroquinocellulose, akasha, boroethi Excipients like lene glycol, mannitol, salt or sodium citrate It is also preferable to add. Preferred compositions for parenteral administration further include liposome carriers. Consists of the compound put into the body. Liposomes contain peptides together with phospholipids , conventional hand shaking, high pressure extrusion, with or without Colesteel. dispersed in the aqueous phase by various methods such as , reverse phase evaporation, and microfluidization. It can be formed by Further details are disclosed in U.S. Patent Application No. 763,484. Ru. Such carriers optionally contain immunoglobulins reactive with virus particles or infected cells. directed to its active site by a brin or protein. Infection as well as protein selection Depends on the antigenic determinants of the virus. Examples of such proteins include human immunodeficiency virus ( CD-4T-cell glycoprotein or 5C reactive with the glycoprotein coat of HIV and its derivatives such as D-4 (soluble CD-4).

These proteins are disclosed in US Patent Application No. 160,463. to other viruses Similar target proteins for the same can also be devised by known methods and are within the scope of the present invention.

Alternatively, the peptides can be packaged in capsules, tablets, emulsions or shells for oral administration. It can be a lop. Enhances, stabilizes, or facilitates production of compositions A solid or liquid pharmaceutical carrier can be added to make the drug. Shironobu for liquid carrier, Included are peanut oil, olive oil, glycerin, saline and water. on a solid support Starch, lactose, carunium sulfate dihydrate, white china clay, pectin, acacia, agar or or gelatin. The carrier is glyceryl monostearate or glyceryl monostearate. Also included are sustained release agents such as lyldistearate alone or in combination with wax. It will be done. The amount of solid carrier will vary, but preferably about 20 mg per dosage unit - about It is 1g. These pharmaceutical preparations are manufactured by conventional methods. When using a liquid carrier , white knob, elixir, emulsion or aqueous or non-aqueous suspension. can be done. Such liquid formulations can be administered directly parenterally or with soft gelatin. It can be administered in capsules.

For rectal administration, powders of the peptides of the invention may be added to cocoa butter, glycerin, gelatin or can be combined with excipients such as polyethylene glycol and formed into suppositories. . Powders can also be used in buffered or unbuffered oil-based formulations, gels, creams or emulsions. It can also be prescribed as a transdermal medication.

The beneficial effects can be seen in combination with the protease inhibitory compounds of the invention and other antiviral agents individually. Alternatively, they can be obtained by administering them together in combination. antiviral agent Examples include nucleoside analogs, phosphonoformates, rifabutin, ribaviran , phosphonothioate oligodeokine nucleotide, castanospermine, Dextran ruphate, alpha interferon and unviridan is included. 2′,3″-dideoxyadenine (ddCL2°.

3°-dideoxyadenine (ddA) and 3′-azido-2′.

Nucleoside analogs including 3-dideoxythymide (AZT) are particularly useful. It is for use. AZT is one of the preferred drugs. Suitably, the pharmaceutical composition is antiviral. the protease-inhibiting peptide of the present invention and a pharmaceutical carrier.

The present invention will be explained in more detail by giving Examples below. It is not limited to these.

In the examples, all temperatures are in °C, and amino acid analyzes were carried out using On (Dionex Autoion) 100 was used. Analysis of peptide content was performed based on amino acid analysis. FAB Mass Nosvector is a fast a VG using Tom Bombardment Performed on Zab Mass Spectrometer It was. The abbreviations representing elution compositions for thin layer chromatography and self-flow distribution are B-n. -Butanol, A: acetic acid, W/water, E: ethyl acetate and IP isopropaz It is a rule. NMR was performed using a Bruke AM250 spectrometer. It was recorded at 25QMHz. Multiplicity is S:/ngrenot, d. t, t-triblet, q: quote, t, m-multibret, t and br, It's a broad signal. 4(S)-amino-3(R)-hydroxy-5- Phenylhexanoic acid (ARPPA) and its 3(R) epimer (3-R-AH PPA) according to the method of J. Med. Chem., 23.27-33 (1980). Manufactured from. The racemate (3-R,5-AHPPA) Manufactured by omitting matoography.

Purification of recombinant HIV protease A method for expressing recombinant HIV protease in E. coli is provided by Proc, Nat. 1. Acad, 5c-i, USA, 84.8903-6 (1987) ing. Enzymes used for analysis of the peptides of the present invention are produced by this method and are as follows: and purified from cell pellets. E. coli cell pellet at 50mMT 5-HCI (pH 7.5), 1 each. QmM DTT, EDTA and P In a buffer consisting of MSF (7E=/l/methylfluoronyl fluoride) Resuspended cells were lysed by sonication and unlysed material was lysed at 15,000 x g for 15 min. It was removed by centrifugation for a while. The supernatant was 40% saturated with ammonium sulfate and this suspension was The suspension was stirred at room temperature for 30 minutes and centrifuged as well. The minimum amount of precipitate obtained is 20 mM Tr i 5-HCl (pH 7,5), 200mM NaCl , each Q, were redissolved/resuspended in 1mM DTT and EDTA. Transfer the sample to Beckma TSK G20005w Breno Late HPLC Gel Filtration Column (21x The column was centrifuged at 4 ml/min with the same buffer. Equilibrated at a flow rate of The column eluate was monitored at 280 nm and fractionated for 1 min. tion was collected. Typically, rHIVPRT (recombinant HIV protease) eluted at 45-46 minutes. At this stage, protease 4tN85-95% pure It is. According to ImmunoProto analysis, more than 90% immunity was achieved in the ammonium sulfate process. Epidemic-reactive substances precipitated. In the activity analysis, the highest activity peaks were at 45 and 46 minutes. was observed in the collected fractions. By RP-HPLC and 5DS-PAGE The analysis of TSK Rikram Urakunyeon shows that most of the 1100 Mr protein is 45 and 4 It was shown that 6 frac/yon was observed. The activity itself is affected by high salt, so i.e., with increased salt, activity levels increase, so reliable recovery data It is not something that gives In this way, higher activity is recovered at each step of purification. Ta. The total acid rate of rHIVPRT was 1 mg N from 50 mg E. fliberette. It was.

Initial rate data for substrate dynamics oligopeptides associated with HIV protease activity are shown on p. Measured at H6, 0, 37°C. The reaction mixture (0,01-0,1ml) is 5Qm M　Me　S　(2-N-mo/l/phorino)ethanesulfonic acid), 1mM EDTA, 1mM DTT, Q, 2M NaCl 1,0,1% (v/v) Lidoa X-100 (pH 6,0), 10% DMSO and various concentrations of peptides Contains de. After several minutes of incubation at 37°C, purified HI The reaction was started by adding zeolite (Olol-2 mg) and after 10-30 minutes, an equal volume of water was added. Cold 0.6N trichloroacetic acid or 0.6N trichloroacetic acid. Quenched with 2% trifluoroacetic acid.

The sample was centrifuged at 17,000 rpm for 5 minutes to remove the peptide substrate and product from the reverse phase. Separation and quantification by HPLC (peak integration). The initial rate depends on the given substrate concentration. Based on the conversion of substrate to product (mM peptide produced) measured in 7 minutes . Typically, the reaction rate is linear with respect to the reaction time, reaching 1 at the end of the reaction. Less than 5-20% of the substrate was converted to product. Kinetic constants (Miberellis constant, maximum speed (Adv, Adv, Enzymol, 29.1. (1967)), Mivaelis-Menten It was determined by substituting it into the equation (v=Vmax(S)/Km+(S)). turn The over number (kcat) is calculated using It is obtained from k c a t = Vmax / E t. Ta.

Evaluation of substrate activity was determined by relative initial velocity (relVo). This analysis is similar to After incubation, the % hydrolysis of the substrate after 10-20 minutes was determined by Ac-Arg-A la-3er-Gin-Asn-Tyr-Pro -Va l -Va l - The same procedure was performed except that the % hydrolysis of NH, (control) was compared. This allows the contrast Comparison of initial reaction rates for peptides at one point in time was possible.

Inhibition of HIV brotease activity A typical assay consists of LOML MENDT buffer (50mM Mes (p H6,0), 1mM EDTA, 1mM dithiothreitol, 200mM NaCl, 0.1% Triton Arg-Ala-3er-Gin-Asn　-Tyr-Pro-Va　IV a I-NH, , Km = 7mM) and submicromolar synthetic substrate.

Incubate for several minutes at 37°C and add 0.01-1 mg purified HIV protease. The reaction was started with ze. After 10-20 minutes, an equal volume of the reaction mixture (37°C) was cooled. Quench with 0°5 NI-lichloroacetic acid, remove the precipitate by centrifugation, and separate the peptide. The solution was subjected to reverse phase HPLC (Beckman Ultrasfy-OD S, 4.5x25r nm, mobile phase, 5-20% acetonitrile/l/HtO-0, 1% TFA (15 minutes), 20% acetonitrile/H, O-Q 1% TFA, 1.5ml/min , detection 220 nm). Ac-Arg-A, 1a-3er-Gln -Asn-Tyr-Pro-Va l V a l N Hx (1718 ) and Ac-Arg-Ala-5er-Gln-Asn-Tyr (10- 11 minutes) was confirmed using a standard product. Ac-Arg-Ala-3er-Gl The initial rate of n-Asn-Tyr formation was determined from the integration of these peaks.

Typically, the inhibitory properties of a synthetic compound are measured by the slope of a plot of l/V versus (inhibitor). /determined from intercept analysis (Dixon analysis). From this type of primary analysis The derived Ki value is determined only by the competitive inhibitor and by the good Miberellis constant of the substrate used. Accurate under known conditions.

Example Peptide amides are solid-phase peptides using benzhydrylamine resin as a carrier. Synthesize by synthesis. Starting from the carboxyl terminus, until the desired sequence is obtained , sequentially adds protected amino acids. For protection of α-amino groups, t-butyl oxychloride A cycarbonyl (Boc) group is used. Side chain functional groups are protected as follows; luginine and histidine, tosyl (Tos), nigistein, p-methyl base serine and threonine, benzyl ether (Bzl) ; Lysine, p-chlorocarbobenzoxy (C1z); Glutamic acid and Spartic acid, benzyl ester (OBzl); tyrosine, p-promocal Pobenzoquine (BrZ). Removal of the E3oc group was performed using 50% truffle in methylene chloride. This is carried out by treatment with fluoroacetic acid (TFA). Neutralization of amine-TFA salts is treated with 7% diisoprohylethylamine (DIEA) in methylene chloride. To do this. Amino acids were mixed with 3 equivalents of Boc-amino acids and 3 equivalents in DMF. of 1-hydroxyhensitriazole (HOBt) in methylene chloride. of dicyclohexylcarbodiimide (DCC) to the growing peptide. do. Completion of ligation was checked by ninhydrin test.

and repeat the concatenation if necessary. The general protocol is as follows.

1. Washing with CH, CI, Washing with 2.50% TFA for IXI minutes txt min 3. Deprotection with 50% TFA lX 20 min 4, CH, C1 Washing at t 8 x 1 min Neutralization with 5.7% DIEA 3 x 2 min 6. Washing with CH, CI, 4×1 minute 7. Washing with DMF 2X1 minute 8, Boc-AA+HOBt/DMF No discharge 9, DCC /CI(, CI, 2 hours 10, washing in DMF # 2 x 1 min 11 washes with CH, CI, 3 x 1 min 1st (C-terminus) If the residue is to be attached to a BHA resin, the synthesis begins in step 5. everything after For the amino acid, synthesis begins in step 1.

Ac-Arg-Ala-5er-Gin-Asn-Tyr-Pro-Vat -Val　-NH, production Protected nonapeptide resin Boc-Arg (Tos)-Ala-Ser (Bz l)-Gln-Asn-Tyr　(BrZ)　-Pro-Val-Val-B) fA was produced by a conventional method. The N-terminal Boc group was added with 50% TFA/CH, CI. After removing and neutralizing the obtained TFA salt with 7% DI EA/CH, CI, Resin-bound peptide was acetyl anhydride for 30 minutes in 2 ml of acetic anhydride/CH, CI, 30 ml. It became.

Treat with 0 ml of anhydrous liquid HFl for 50 min at 0 °C in the presence of 1 ml of anisole. The peptide was cleaved from the resin by removing the side chain protecting group. under vacuum, After removing the HF, the resin was washed with ethyl ether and air dried. Then, The resin was washed with 2 x 30 ml of 1% HOAc/H,O followed by 2 x 30 ml of 1% HOAc/H,O. Extracted with 0% HOAc/H,O. The combined extracts were lyophilized to obtain crude peptide 4. 76 mg was obtained.

The crude peptide was treated with n-BuOH/HOAc/H1O (4:1:5) system. It was purified by the run-of-the-mill distribution method. Pool the appropriate fractions, evaporate to dryness, and remove the residue. The partially purified peptide was lyophilized from 1% HOAc/HtO. 294 mg was obtained. A 100 mg portion of the partially purified peptide was further added to the eluate. 2゜6X70cm G-15 Cephadex (S) using 1% HOAc as Purification was performed by gel filtration on an ephadex) column. appropriate fraction They were pooled and lyophilized to yield 89 mg of the purified title compound.

TLC: (n BuOH/HOAc/H=O (4: l: 1)) Rr =0°49; HPLC: (Hamilton PRP-1, CH, CN/Hto10.1% TFA, 5% to 40% CH, CN, 15 minutes, linear graph Gent, 1.5ml/min) k'=2.6; FAB-MS: m/z 10 74 (\44-H”).

Example 2 (2S)-N-(2-(N-tert-butyloxycarbonyl)amia) L-phenylalanine methyl ester w acid ito,og (464 mmol) , 2.9 ml (92.8 mmol) of triethylamine I and 60 DMF It was dissolved in ml of compound a. N-t-butyl dicarbonate 16.0ml (6 9.6 mmol, 50% excess) was added over 5 minutes at room temperature. Stir for 16 hours After stirring, the mixture was filtered and concentrated under reduced pressure. 20% EtOAc/hexa Fluff 5 chromatography on silica gel using 13.0g was obtained.

TLC: (20% EtOAc/hexane) R, = 0.31; NMR (C DC131: δ7.23 Tm, 5H),! 1.16 (d, LH, J-8H z), 4.57 (q, IH, J=7) 121/3,66 Lsz3HL ball 07 Td, 28. J-6Hzl, 1.40 (s, 9H).

N-terL-phytylone carbonyl-phenylalaninal b) the above Dry 2.9 g (46.4 mmol) of manufactured Boc-Phe-OMel It was dissolved in 30ml of toluene and cooled to -78°C. Hydrogenated diisobutylaluminum (25%/toluene, 77.3 ml, 116 mmol, 7 mol, 150% excess ) was added over 2 minutes. After stirring for 15 minutes at -78°C, methanol 15 ml of water was added slowly to control foaming. The mixture was mixed with Rochelle salt ( Rochelle 5alt) (potassium sodium tartrate 94g/liter of water ) and shaken with 100 ml of ether until extracted.

The aqueous phase was washed with ether (3X 100ml). The organic phases were combined and dried ( ＼−1g5　O+) and made it emit a sound.

The product obtained was used directly in the next step.

TLC: (20% EtOAc/hexane> Rt=’v, 2t; NK R (CDC13): δ9.69 <3. IH), 7.28 (rn, 514 1.5.05 +5. IH), 4.43 (q, LH, J-7Hzl, 3.1 0 (d, 2H, J-7Hzl, 1.43 (s, 9H1.

(2S) N (2-(N-tert-butyloxycarbonyl)ami/- 3-phenyl)propyl-L-proline benzyl ester C) 2 (b) above 18.1 g of L-proline benzyl ester hydrochloride ( 75 mmol) in 250 ml of 1% acetic acid/methanol. cyan Add 3.06 g of sodium borohydride (48.7 mmol, 5% excess) , the mixture was stirred at room temperature for 16 hours. The solution was then evaporated and the residue was Partitioned between tOAc and IN hydrochloric acid. The organic layer was treated with IN hydrochloric acid (2x), IN a Washed with HCOo (3X) and water (IX). Evaporate the organic phase and 13.5 g of the above benzyl ester was obtained. Flash chromatograph some of the crude material The pure product was purified by Raffy (silica gel, 20% EtOAc/hexane). Get it, CHC it! , and crystallized from hexane. These crystals are satisfying The X-ray crystal structure (S, S) was shown.

Melting point near 9-79.5°C.

TLC: (20% EtOAc/hexane) R, = 0.31; NMR (CDC1 3): δ7.33 (s, 5H), 7.20 (s, 5H), 5.14 (s, 2H), 4.95 (d, LH, J-9Hz), 4.05 (d , IH, J-7Hz)t 3.17 (m, 2H1,2,63(m, 514 1゜1.87 (m, 4H), 1.35 (s, 9H) - MS: m / Z 439 (M!H)”, 383.365.347.339.303 ．． 231.218.91.79゜Elemental analysis: CtsH34N, O, as Calculated value (%): C, 71,21; H, 7,81; N, 6.39 Measured value (%): C ,71,00:H,7,83,'N,6.25d) Protective reduction of the above 2(c) production Dipeptide 0.44g (1.0 mmol) was added to 10% pd/C (0,50 g) together with 100 ml of methanol and hydrogenated for 1 hour under 45 psiH. Added. The mixture was filtered and concentrated to give 0.35 g of 100% ) get TLC: (B:A:W (4: 1: 1)) R1-0,66; NMR (DM SO-d6): δ7.26 (s, 5H), 6.80 Td, IH, J-9 Hz), 6.00 (s, 281°3.78 (s, IHI, 2.9 (m, 6H1,1,85 (m, 4H1,1,30 (s, 9H).

Using benzyl piperidine-2-carboxylate instead of proline benzyl ester Perform the same operation except that (2S)-N-(2-(N-t-butyloxycarboxylate) Rubonyl)amino-3-phenyl)propyl-piperidine-2-carboxylic acid was obtained. Ru.

Δ3-dehydroproline methyl ester instead of proline benzyl ester Methyl (2S)-N- (2-(N-t-butyloxycarbonyl)ami/-3-phenyl)propyl -L-Proline is obtained. The methyl ester was dissolved in methanol:water (4:l) at 5% At the same time, it was saponified with Co to give (2S)-N-(2-(N-t-butyloxycarboxylate) 7-3-phenyl)propyl-L-proline is obtained.

Implementation M3 (IR,2R)-2(((2S)-2-tert-butyloxycarbonylamine Nor-3-phenyl-1-oxo)propyl)fucloben 6-carpomethoxypi9 2 mouths [3,1,0] Beef San a) Ethyl diazoacetate 53ml (0,50 mol ) in a 20-25°C water bath for 4 hours via a syringe device. 1.0 g (0,0023 mol) of rhodium dihydride and 250 cyclopentene m1 (25 mol) was added to the stirred mixture. After 30 minutes, excess 7 clovente The chlorine was removed using a rotary evaporator.

'f<　if diluted with CH, CI, and filtered through Celite. and concentrated on a rotary evaporator. Residue at 44-46°C 10°5mm Distilled with F (g, yield 6o%) as a mixture of carboetquin epimers. 46.5 g (0.302 mol) of the compound was obtained.

TLC: (hexane:ethyl acetate (80:20)) R, ~0.67.

NMR (CDC13): δ4.1 (2H, good @t4t) 2.0 -1.5 ('7H, m), 1.45-1.1 (5H, ml.

6-(1-hydro+7-1-methylethyl)bi/ri 0 [31°0: Hegyu San b) 1,4 M solution of methyllithium/ether (350 ml, 0.50 mol) The solution was stirred rapidly at −78° C. under argon to produce the product 2 of Run 1Te13(a). 7.67 g (0,180 mol) was added via syringe over 15 minutes. . The mixture was stirred at -78°C for 1 hour and then allowed to stand at room temperature for 2 hours. Warmed to warm. Add 400 ml of water, then 3N hydrochloric acid (approximately 200 ml) to the solution. The solution was carefully added until it was neutral. Separate the ether layer and dilute the aqueous layer with ether. Extracted twice. The combined ether extracts were dried with MgS O4 and rotary -Concentrated using an evaporator. Distill the residue (80-90”C/15mmHg) , 19.79 g (0,141 mole) was obtained.

TLC: (hexane, ethyl acetate (80220)) R, ~0.36, O17° NMR (CDC13): δ2.0-1.6 (5H, ml, 1.5-, 1. 0 (9H,'m+, 0.55 flH.

ml.

<=>-trans-2-(2-methylpropenyl)chlorocycloc)THF 1 A solution of 19.6 g (0,140 mol) of the product of Example 3(b) above in 23 ml of 250 ml of 37% aqueous hydrochloric acid and 125 ml of THF cooled in a water bath. Added all at once to the rapidly stirring mixture. After 3.5-4.0 minutes, the mixture was poured into ice water (1 liter). The mixture was immediately extracted with pentane (3×230 ml). The pentane Wash the extract with 5% NaHCO’ + 200ml and dry over magnesium sulfate. It was concentrated using a rotary evaporator (25° C./15 mm). The residue, 78-b through an 8-inch Vigreux column 18.65 g (0,118 mol) of the title compound was obtained.

TLC: (hexane) R, = 0.71; NMR (CDC13): δ4.95 ? lH, dil, 3.84 (IH, Akira 5g%rJ - Jujujuku).

ml.

GC-MS (isobutane): m/z 158 (M)', 123 (M+H-HC I)’; IR (film layer)・2980.1455.1382.838cm”.

2-(2-methylpropenyl)lithiocyclobencune d) Equipped with a reflux condenser The resulting dry 50 ml flask was purged with argon. Powdered lithium 0.34g (4 9 mmol; 1% N aalt; 25 wt% dispersion/mineral oil) and dry pentafluoride. (Rinse with 2XlOm to remove mineral oil. Dry freeze-thaw and degas In 20 ml of treated pentane, 1.58 g of the product of Example 3(C) (10, A solution of 0 mmol) was added. The mixture was stirred and refluxed for 19 hours in an oil bath at 55°C. and then allowed to stand. The supernatant lα was collected via a syringe and dried under argon. Filtered through a dry fritted glass filter. The resulting clear yellow solution 17.6 mt for menthol using 1,10-phenanthroline as indicator Titration showed 0.45M in the title compound. 79% yield of title product I got it.

e) Boa-L-phenyl in 40ml ether at -78°C1 under argon. To a stirred solution of 1.59 g (,6,00 mmol) of alanine was added n-butyllithium. M4. OOml (1,50M/hexane; 6.00 mmol) was added . After 10 minutes, 37 ml of the product of Example 3(d) (0.32 M/pentane: t, t , a mmol) was added dropwise over 10 minutes. After 10 minutes, the solution was brought to O'C. Warm and stir for 1 hour and 45 minutes, then add 200 ml of water with vigorous stirring. Ta. The mixture was extracted with ethyl acetate and the crude product was flash chromatographed. Purify the product three times with Hexane/ethyl acetate (9525) and elute first. Isomer A (0,334 g, 8.46 mmol; yield 14%) is then eluted. 80.356 g (9.60 mmol; based on Boc-L-Phe) Yield 16%) was obtained 7:.

(isomer B) (IR, 2R) 2 ((2S) 2-tert-butyloxycarbonylate Ruami/-3-phenyl-1-oxo)propyl-2-methylpropenylcyclo Benkun TLC: (hexane: ethyl acetate (95: 5)) R, = 0.19: NM R (CDC13): δ7.2B-7.08 (5H), 5.06 (1B, d; J-1,8Hz; NHI.

4.95 (IH, d; J-9, 6Hz; vinyl tJ), 4-61 +IL Akira bv-ri tnv nishiki L3.11 (IJ m)t 3.1 0 (IHr ad; be>’, :’+L-4L)/2.83 (LH, dd; Be>Zure 41.2.77 (1M, ml, 2.05 -1.25 (6H), 1.68 (3H; butyl).

1.62 (3H, / child IL,), 1.39 (9H, s; B ocl.

MS (DCI, / N H3), + m / z 372 (M + H )’, 316.2 (isomer A) (IS, 2S)-2((2S)-2-tert-butylox7carponyl Ami/-3-phenyl-1-oxo)propyl-2-methylpropenylcyclobene Nkun Melting point, 42-44°C0 TLC: (hexane: ethyl acetate (95: 5)) R, = 0.26: NMR ( CDC131: δ7.27-7.08 (5H), 5.18 (IH, d; J-7,7Hz; NHI, 5.00 (IHt d; J79.3 H 2N e’2 + w), 4.51 IIHt wq et al. prystm), 3.05 (IH, dd; J-13,9,6,7Hz; s>’ν+’ L) / 2.95 +IH, dd; J-13,9,5,W H2; Pe>VoJt>, 2.79-2.58 (2H, m), 1.95- 1.20 (6H, ml, 1.70 (3H; +IL-L 1.52. (3H; x++!-L L, 4j (9H; Boc)-MS (DCI /NH3): m/z　　　(M　H)", 372.316, (IR, 2R)- 2((2S)-2-tert-butyloxycaf)CH=CI-/CH30H (4:1) in 5 ml, 186 mg of the product of Example 3(e) isomer B (0,5 0 mmol) solution at -78°C until the blue color persists (approximately 2 minutes). ), an ozone flow was introduced. The solution was purged with argon and (CH3),S(0, 5 ml) was added and the solution was warmed to 20'C. 1 & volatile components under vacuum for 2 hours It was removed. The obtained crude altehyde was dissolved in 5N]1 acetone and added to o’c. While stirring for 3 minutes, add Jones Reagent 0, It was reacted with 25ml (2.0M). Added 02ml of isopropanol . The mixture was diluted with water, extracted with CH, C1, and the extract was concentrated. flash Shuchromatography (hexane:ethyl acetate (80:20), followed by Hexane: ethyl acetate: acetic acid (80:20:4)), the yield was 82%. 148 mg (0,410 mmol) of the title compound was obtained as a sliver crystal.

Melting point: 9-4.5-96°C.

TLC: (hexane: ethyl acetate: acetic acid (80: 20: 4) Rr=0.2 7: NMR (CDC13): δ7.26-'7.13 (5H, ml, 5.12 ( IH, d; J-8,0Hz).

4.70 (IH, clear v+5 1” 111 yen) / 3-46-3-27 (2Hz In) t 3-16 +18/ ddG J"14 Hz, 6.2 Hz), 2.93 (IH, dd; J -14.0Hz, 6.5Hz), 2.14-1.67 (6H, ml , 1.39112H,s).

MS (DC, I/NH,): m/z 362 (M+H)”3(a) Smell The same procedure is used, except that cyclohexene is used instead of cyclopentene. 2((2S)-2-tepropyl)cyclohexanecarboxylic acid get.

Example 4 (Is, 2S) Up to Himee μs upper two legs ± Production of xo-3-phenylfurovir)cyclobentanecarboxylic acid 186 mg (0.50 mmol) of isomer A of Example 3(e) is used instead. Except for this, in the same manner as in producing the compound of Example 3 (1), the yield was 75%. 35 mg (0.37 mmol) of the title compound was obtained as a white solid.

TLC・(hexane:ethyl acetate:acetic acid (80:20:v)Rt=3. 08 (2H, ml, 2.91+IH, dd; Jm13.8.6.9 Hz; &>:;zwa)t 2.20-1.62 (6H, ml, 1.38 (12 H,s).

MS (DCl/'NH3): m/Z 362 (M→-H)°, 3 23 (M-i-NH1+H-t Bu)', 262.120 (IR, 2R)- 2-(((IH3,2S)-1-hydroxy-2-tert-butyloxycal Production of bonylamino-3-phenyl)propyl)cyclopencune carboxylic acid 216 mg of the compound of Example 3(f) in 5 ml of methanol at 0°C (0.5 98 mmol), stir NaBH + 25 mg (0.66 mmol). I added it while doing so. After 20 minutes, dilute hydrochloric acid was added and the mixture was extracted with CH, CI, The title compound 21 as a mixture of epimers in a yield of 1OO% and a ratio of about 2:1. 6 mg (0,595 mmol) was obtained.

NMR, (CDC13): δ7.35-7.15 (5H), 5.05 ( 1) 17 NHI, 3.87-3.56 (214).

2.95-2.60 (381, 2, 38 (IM), 2.19-1.55 ( 6H), 1.38 (9H).

MS (DCr/NHs) + m/z 364 (M+H)', 325,308 (MC(CH,), +H)', 264 (M OCOC(CH3)3+H )", 183, 160, 120 2-(((2S)-2-tert-butyloquine/, -3- The same reduction for phenyl-1-oxo)propyl)cyclohexanecarboxylic acid Perform the original operation to obtain the corresponding 2((2S)2-tert-butylox7carponyl Amino-3-phenyl-1-hydroxy)propyl)cyclohexanecarboxylic acid get.

a) 140 mg of the epimer of Example 5 (0,386 millimolar) solution with excess ethereal diazomethane for about 5 minutes (until the yellow color persists). ). Acetic acid (0.1 ml) was added and the solution was evaporated on a rotary evaporator. It was concentrated in a Hpt on silica, c (vinegar fi ethyl: to beef san (80: 20)), the first eluting isomer A was 47.0 mg (0.125 mg). 850.6 mg (0,134 mmol) of the isomer was obtained.

(isomer A) (IR,2R)-methyl-2(((Is,2S)-2-tert-butyloxy (cycarbonylamino-1-hydroxy-3-phenyl)propyl)cyclopenta Carboxylate TLC: (hexane:ethyl acetate (2:I)) Rr=0 ．． 54: NMR (CDC13): δ7.32-7.18 (5H, +t+l , 4.85 (IH, d; J-8, 7Hz; NHI.

3, 2 (IH, ml, 3.66 +3H, sl, 3.52 (IH, dd), 2.90 (2H, Ill et al. d; S-shi’c41L H, 2, 58 (IH, Q6tt’J E direct fi 1. l, 2.40 (1M, ml.

1.92-1.50 (6H, m), 1.39 (9H, sl.

MS (DCI, 'NHs) + mlz 378 (M+H)", 339.3 22 (λI-C(CH3), +H)', 321,304 (M-QC(CHl) , 〒H)゛, 120 Fourteen 2-hydroxy configurations were given as follows. Reacting isomer A with TF A to remove the Bo4 protecting group, followed by reaction with phosgene and triethylamine. It was transformed into its oxazolidinone derivative by The oxazolidinone ring Hydrogen has an NMR coupling constant (J = 5.3 Hz) consistent with the trans function 1. , which is confirmed by the lack of observed NOE, and thus the 2-hydrocarbon It was confirmed that the roxy configuration was (S).

(isomer B) (IR,2R)-methyl-2-(((IR,2S)-2-tert-butyl xycarbonylamino-1-hydroxy-3-phenyl)propyl)cyclopene Tancarboxylate TLC: (hexane:ethyl acetate (2:1)’): R, = 0.42°NMR (CDC13): δ7.31-7.16 (5)! , m), 4.92 (IH, d; J-, 8,7Hz), 3. W 9 (IH, m), 3.72 (3H, s), 3.64 (IH, sl et al *’i -d) t 3-41 (IHldd; JI-14-2r 4-' 2 Hz r ■-Lylic 1, 2.72 (2H, ml, 2.36 (11, T IFI et al. p9" 141 z 2.06-1-40 (6H, ml / X, 3 2 +9H, s) -MS (DC engineering/NH31: mlZ 37B (M+H1 ”, 322t 278-120° The 2-hydroxy configuration of isomer B is similar to that of isomer A. Measured using the same method.

Alternatively, the two epimers can be analyzed by preparative HPLC (Lynin Dynamax). Rainin Dynamax) 1 in, X 25 cm Silica Column, 20ml/min, hexane:ethyl acetate; formic acid (73:25:l) ), isomer A (yield 49%) and isomer B (yield 35%). %) was obtained.

(IR, 2R)-2-(((Is, 2S)-1-Hydroxy-2-tert-B Tyloxycarbonylamino-3-phenyl)propyl)cyclopentanecarbo hmm b) The isomer of Example 6(a) in 2 ml of THF:water:methanol (2:1) To a solution of 23.7 mg (63.2 mmol) of body A, add 10% NaOH (approximately 0.2 mmol) was added. 2. ．． After 5 hours, the mixture was diluted with ethyl acetate and 5% salt Washed with acid. M　g　dried with S　Oa, evaporated and marked with a yield of 94% 21.4 mg (59.0 mmol) of the compound was obtained.

Melting point: 120-122℃.

TLC: (hexane: ethyl acetate: acetic acid (80: 20: 4) R, = 0.3 4゜ NMR (CD30D) 2 δ7.23 (5H), 3.73 (IH, ml,, 3.53 (LH, ml, 2.85 (IH, dd; 4.i), 2.73 +IH, dd; Be successful), 2. 55 (LM, □).

2.37 (IH, ml, 1.90-1.57 (6H), 1.35 (9 H,s).

Example 7 (lR,2R)-2-(, ((IR,2S)-1-hydroxy-2-Example 6( Following the procedure in b), 1 mg (53.6 mmol) of isomer B of Example 6(a) was added. 19.3 mg (53.6 mmol; yield 99%) of the title compound was obtained.

Melting point 133-135°C0 TLC: (hexane: ethyl acetate: acetic acid (80:20:4): Rr=0.27° (2B), 1.50 (IH, m), 1.30 (9H, s>Example 8 (IS,2S)-2(((IH3,2S)-2-tert-butylox7carbo nilami/-1-hydroxy-3-phenyl)propyl)cyclopentanecarbo production of phosphoric acid Example 5 except that 38 mg (0,105 mmol) of the compound of Example 4 was used. Following the procedure, the title compound 35 was produced as a white solid in a 1:1 ratio and 93% yield. ．． 5 mg (0,0978 mmol) was obtained.

TLC: (hexane: ethyl acetate: vinegar fl! (80: 20: b) R r=0.26; NMR (CDC13): δ7.32-7.14 (5H,', ml, 3.93 -3.60 (2H, ml, 3.38-3.21+IH, 2>, *fyi a), 3.05-2.70 (2H, ml, 2.55-2.38 (IH, +n l.

2.35-1.54 (7J m) t l-31-1,14 (9H+ 3- s-1# & )-a) Isobutyl chloroformate 1.30ml (10.0ml) N-Cbz-L- in 30 ml of THF at -40°C under argon. 2.51 g (10.0 mmol) of valine and 1°21 of N-methylmorpholine m1 (11.0 mmol) over a period of 10 minutes. 1 After hours, solid L-valine methyl ester hydrochloride 1.68 g (10,0 mmol) and then add 1.21ml of N-methylmorpholine (11.0ml). mol) was added. The mixture was stirred for 24 hours while warming to 20°C, then filtered. and concentrated to a solid using a rotary evaporator. flash chromatography Gradient, 0-5% CH3OH/CHCl, yield: 9 3.44 g (9.50 mmol) of the title compound were obtained as a white solid at 5%.

NMR (CDC13): δ7. j4 (5H, ml, 6.80 (IH, d), 5.62 (1M, d), 5.13 (2M.

s), 4.56 (IH, dd), 4.17 (LH, ddl, 3.74 ( 3H, s), 2.15 (2H, m).

1.00-0.88 (12M, weight 71 2' [4).

Valylvaline methyl ester tiKli [b) of Example 9(a) in 4 ml of acetic acid. A solution of 535 mg (1.47 mmol) of the compound was added to 10% Pd under a hydrogen atmosphere. /C (50 mg) and stirred for 5 hours. The mixture was filtered and diluted with CH,OH. The mixture was diluted, acidified with 0.2 ml of 37% hydrochloric acid, and concentrated to a gum under vacuum.

The residue was triturated with ether and dried under vacuum, labeled as a white powder. 395 mg (1.47 mmol) of the compound was obtained.

NMR (CD30D): δ4.36 (IH, d; J-5,8Hz), 3. 81 (IH, d; J-5,7Hz).

3.73 (3H, s), 2.21 (2H, 21g), 1.09 (3H, d;, 7-7.0 Hz), 1.06t3H,a;, 7-6.9H z), 0.998 (6J(, *?l = L4?, ).

MS (FAB 1VE): m/z 231 (M+H)', 188.171. 132.72.55 Example 10 tert-butyloxycarbonyl-(0-phenylmethyl)serylalanilua Production of lanin tert-butyloxy group (0-phenylmethyl)serylalanine chill ester a) 2.59 ml (20.0 mmol) of isobutyl chloroformate was heated with argon. Bottom, N-Boc-L-serine-O-benzi in 100 ml of THF at -40°C. ester 5.90 g (20.0 mmol) ester N-methylmorpholine 2. It was added dropwise over 15 minutes to a stirred solution of 74ml (25゜Ommol). . After 45 minutes, 2.75 ml of N-methylmorpholine (25.0 mmol) and and 3.22 g (23.0 mmol) of solid L-alanine methyl ester hydrochloride. added. The mixture was stirred while gradually warming to 20°C. After 18 hours, the mixture The mixture was diluted with ethyl acetate and washed sequentially with 5% hydrochloric acid, 5% NaHCO, and brine. Washed thoroughly. Filter and remove the solvent under reduced pressure to obtain an oil in 99% yield. 7.54 g (19.8 mmol) of the title compound were obtained.

TLC: (Ethyl acetate Nihegyu San (1:1)) Rr=0.5: NMR ( CDC13): δ7.32' (5H, ml, 7.1lT1H, brm), 5.11 (IH, br ml.

4.65-4.53 (3H, m), '4.30 (IH, br ml, 3.92 (IH, dd; J=9.2 Hz, 3.X Hz), 3.73 (3B, s), 3.58 (1M, dd; J”9.2 Hz, 616 Hz+, 1.45 (9H, 5ly1.39 (3H, d; J -7, iHz).

b) In 50 ml of water, add 0.8 g (20 mmol) of sodium hydroxide to methanol In 225 ml of water (2:1), 7.5 g (19 , 7 mmol) portionwise over 2 hours. The solution is approximately neutral. Stirring was continued for approximately 1 hour more until TLC indicated the reaction was complete. Loosen the solution. It was concentrated using a tally evaporator, diluted with water, and extracted with ethyl acetate. water The layers were acidified with 10% hydrochloric acid and extracted twice with ethyl acetate. Mg5O, dried over and concentrated in vacuo to give the title compound as a glass in 96% yield, 6.89 g. g (18.8 mm mono-) was obtained.

NMR (CDC13): δ7.32 (5H, m), 5.62 (LH, >o-t”L 4.60 (lH, ml+4.55 (2M, +1) , 4. ．． 36 (IH, 7-CI-)', 1.3.87 (1)! , dd), 3.61 (kH, ddl.

1.46-1.40 (12H, m).

tert-butyloxycarbonyl-(0-phenylmethyl)serylalanilua Ranin methyl ester C) 2.42 ml (18.8 mmol) of isobutyl chloroformate was heated with argon. Below, compound 8.89 of Example 10(b) in 100 ml of THF at -40°C. g (18,8 mmol) and N-methylmorpholine 2.74 ml (25 , 0 mmol) over 18 minutes. After 20 minutes, N-methy 2.74 ml of lumorpholine (25°0 mmol) and solid I,-alanine 3.07 g (22.0 mmol) of methyl ester hydrochloride were added. 2 of the mixture Stir for 20 hours while gradually warming to 0'C, then dilute with ethyl acetate. ．． Washed sequentially with 5% hydrochloric acid, 5% NaHCO, and brine. filter and reduce The solvent was removed under pressure to obtain 7169 g of crude product as a solid.

Recrystallize (hexane: ethyl acetate (2 L)) to obtain white crystals with a yield of 89%. 7.55 g (16.7 mmol) of the title compound were obtained.

TLC: (ethyl acetate:hexane (tit)) R, = 0.15; NMR (CDC 13): δ1・, 32C5H, mlt 7.00’ (1M, d), 6.9 0 (IH, d), 5.44 (1M.

dl, 4.58 (2H, s), 4.54 (2H, ml, 4.30 (IH, 'y-o-pi), 3.90 (IH, ddLd) Sodium hydroxide 0.25g (6.2 mmol) in Example tO A solution of 2.79 g (6.19 mmol) of compound (C) was dissolved at pH was added in small portions at a rate such that the temperature was maintained between 7 and 9. Stir for another 10 hours. The solution was then concentrated using a rotary evaporator. add water Then, the solution was extracted twice with ethyl acetate. The aqueous layer was acidified with 10% hydrochloric acid and Extracted with chill three times. The organic extracts were dried (MgSO4) and the solvent was removed under vacuum. 2.58 g (5.90 mm) of the title compound was removed as a white powder with a yield of 95%. mole) was obtained.

NMR (CDC13): δ7.30 +78. m; 71% +NH), 5.57 +LM, 7. o-j’L 4.64-4.46 (4)! , ml, 4.38 (IH, ?10-)”’, ), 3 ．． 86 (IH, dd), 3.65 (1M.

dd), 1.44 (9H, sl, 1.39 (6H, @ 1 cup per book).

MS (FAB+VE): m/z 438 (M+H)', 382.338. 293.249.221, 203M5 (FAB-VE): m/z 436 (M-H)-,336,2 Example 11 trans-2-((1-methoxy-1-(2-phenyl-1-phenylmethoxy Production of carbonylamino)ethyl)phosphinyl)cyclopentanecarboxylic acid (±)-2-phenyl-1-phenylmethoxycarbonylaminoethylphosphone acid, monomethyl ester (a)(=)-diphenyl-(2-phenyl-1-phenyl) Synthesis of nylmethoxycarbonylamino)ethylphosphonate (Sy phosphorous acid), 985 (1979). triphenyl (0.1 mol), (freshly distilled) phenylacetaldehyde ( 0.15 mol), benzyl carbamate (0.1 mol) and glacial acetic acid (15 ml ) mixture was stirred for 1 hour until the exothermic reaction subsided. Then, the mixture was Heat at 0-85°C for 1 hour and remove the volatile products under reduced pressure in a boiling water bath. was removed using a rotary evaporator. Dilute the oily residue with methanol 180 ml and allowed to stand at -10°C to crystallize. 1-3 hours later, crystal esthetics Collected by filtration, dissolved in a minimum amount of hot chloroform (30-40 ml), 4 (@ volume) of methanol was added to recrystallize.

A portion of the obtained diphenylphosphonate, 2.44 g (5,00 mmol), was 1.6 g (30 mmol) of sodium methoxide and 75 m of methanol 1 for 2 days. The solution was diluted with 250ml of 5% hydrochloric acid and extracted with ether. I put it out. The ether layer was extracted with 5% sodium carbonate and brine, then concentrated. Shrunk. Ffl12 dimethylphosphonate was dissolved in 30 ml of methanol and 10 ml of methanol. Stirred as a solution in 50 ml of % sodium hydroxide for 1 day. The solution was diluted with 10% hydrochloric acid. Acidified with 300 ml and extracted with C) (, CI). Removal of solvent gave a solid; It was dissolved in 100 ml of hot chloroform. Add 50ml of hexane and add the solution. was stored at -20°C overnight. The obtained crystals were collected by filtration and the yield was 88%. 1.53 g (4.38 mmol) of the title compound was obtained.

2-(2-methylpropenyl)cyclobenthylmagnenium bromide b) Reflux condenser 15 ml of dry ether and a dry vent under argon in a flask equipped with a Stirred suspension of 432 mg (18 mmol) of magnesium shavings in 5 ml of Zen Then, 1.55 ml (18 mmol) of 1,2-dibromoethane was added over 30 minutes. It dripped.

1 part (9.0 mmol) of the obtained clarified MgS Oa solution was added to the pellet. 2-(2-methylpropenyl)lithio in 720 ml of water and 20 ml of THF. -78 of cyclopentane (compound of Example 3 (d)) (8.6 mmol) The resulting heterogeneous mixture added to the °C solution was warmed to O'C and the corresponding Grignard Obtain a clear electroconductive solution of Hall reagent (8.6 mmol) and cool it again to -78 °C. , used in step (b).

trans-2-(1-methoxy-1-(2-phenyl-1-phenylmethoxyca) (rubonylamino)ethyl)-phosphinyl-2-methylpropenylcyclopenta hmm c) Compound of Example 11(a) in dry CHtCl*15mI under fulvone To a solution of 1.92 g (5.50 mmol) of SOCI, 0.40 ml (5.5 mmol) of , 5 mmol) was added. After 2.5 hours the solvent was removed under vacuum. Dry C Add 10 ml of HtCI*1, ”), then add ml of dry THFIO and re-evaporate. The remaining hydrochloric acid was removed. The remaining phosphonyl chloride gum was dissolved in dry THF. 11. Dilute to a volume of 8.6 ml (4.3 mmol) of the solution. ) of the Grignard reagent (8.6 mmol) prepared in step (a) above. Added to the 78°C solution with stirring. The homogeneous mixture was then heated at -78°C for 1 hour. The mixture was stirred at -20°C for 1 hour. 50 ml of aqueous 10% hydrochloric acid, followed by 200 ml of water. m1 was added. The mixture was extracted three times with ether. The combined ether extract was Rinse with % NaHCO, and add fgso.

It was dried and concentrated to Ura (2.1 g). Furanone chromatography (CH Cl,) in poppy, as a gum and as a mixture of four noastereomers. 864 mg (1.90 mmol, yield 44%) of the title compound was obtained.

TLC: (CHC1,:cH,OH(25:1))R,=0.4;GC (CHROM PACK) 51rnmX0゜22mm1 ．． d, ;Liquid phase CPs i I 5CB, oven = 240°C): RT (rat , area) 4.04 (1,0), 4.17 (1,3), 4.42 (l, 2), 4.58 (1, 9) NMR (CDC131: δ7.35-7.03 +10H1m)t 5.56- 5.10 (IHz m; NH) 15.05-4.87 (3H, ml, 4 ．． 33 (IH, ml, 3.76-3.60 (3H, ml, 3.29- 2.69 (3H, ml, 2.08-1.56 (12H, ml, 1.29 (1B,　m).

MS (DCI/NH3): m/z 456 (M+H)", 378.36 5.348.322.213.203.120.108.91 Tanthane carboxylic acid d) Ozonolysis operation of Example 3(f) and compound 68 of Example 11(c) above 0 mg (1.49 mmol) was used to obtain the title compound as a crude gum. centre Rush chromatography (CHCI.

:CH,OH (25:1), followed by CHCl, :CH,OH:acetic acid (25: 1:l)) to give the pure product 36 in 55% yield as a Crisby foam. 5 mg (0,820 mmol) was obtained.

NMR (CDC13): δ7.32-'7.12 (10M, m), 5.0 0-4.91 (2H, m), 4.45+LH, ml, 3.78-3.64 (3H, m), 3.35-2.68 (41, ml, 2.08-1.60 (7H.

ml.

4-bromo- instead of 2-(2-methylpropenyl)lithiocyclopentane Using 1-butene, the corresponding 2-((1-methoxy/-1-(2 -phenyl-1-phenylmethoxycarboxylamino)ethylphosphinyl) Obtain propanoic acid.

Production of bovine sanoic acid to 6-phenyl a) Magnesium powder 1.82 in 10 ml dry diethyl ether under argon A few drops of 4-bromo-1-butene were added to a stirred suspension of g (75 mmol). Ta. At the same time as starting the reflux, 4-build at a rate that maintains a gentle reflux. 6.75 g (50 mmol) of lomo-1-butene was added dropwise. The in compound is heated to reflux. The mixture was stirred for an additional 30 minutes under continuous flow. After cooling, the Grignard reagent solution was (Boc)-L-phenyla in diethyl ether lOmI, dried at °C. Lanin-X-methoxy-N-methylamide 1. Stirring of Og (3.2 mmol) added to the solution. After 1.5 hours, the reaction mixture was diluted with 10% hydrochloric acid and diluted with diethyl ethyl ethyl ether. The mixture was extracted three times with ether. The organic layer was diluted with 1M hydrochloric acid, saturated aqueous sodium bicarbonate, saturated aqueous chloride), washed successively with IImium chloride, dried (MgSO4) and dried under reduced pressure. The residue was evaporated to give white crystals, giving 780 mg (yield: 81.8%) of the title compound.

NMR (CDC13): δ7.20 (5H, ra); 5.73 (IH, m); 5.1 (IH, dl15-05 (2Ht ml); 4-55 (IHr m* a); 3.05 (”-m); 2.48 (2)!, E 14 ); 2.30 (2H, ml; 1.40T9H, s).

MS (DP/NH3): m/Z (M+H)'304; 248.2 0b) Ozone CH,CI at -78<0>C until blue color persists.

400 mg of the compound of Example 12(a) in 5 mI of +MeOH (4:I) (1, 32 mmol) of solution 1α. The solution was purged with argon for several minutes, then Then 2 ml of dimethyl sulfide was added. The mixture was warmed at room temperature for 2 hours and then Concentrate to an oil under reduced pressure.

The oily residue was dissolved in acetone and cooled to O'C while stirring. Johns ( Jones reagent (1.32 mmol, 2M solution) was added dropwise.

After 5 minutes, Improper Tool was added. The mixture was diluted with water and dichloromethane. Extracted three times. The organic layer was dried (MgSOs), filtered, and concentrated to an oil. . Flash chromatography on glue gel (hexane:ethyl acetate (60 :40), then hexane:dityl acetate:acetic acid (65:35:5)), and the oil 175 mg (yield 41%) of the title compound was obtained as a solid.

NMR (CDC131: δ7.24 (5H, ml; 5.1 (IH, d)'; 4 ．． 55. (1M,' w TLIJk I; 3.05 (2H, m); 2.68' (4M, m); 1.38 (9H, sl B MS (FAB'): m/z 322; (FAB-): m/z 320 Using Boc-phenylglycine instead of Boa-(0-benzyl)tyrosine , following the same procedure, 4-oxo-5-(jert-butyloxy/carbonyl alcohol Mino)-5-phenylpentanoic acid is obtained.

Example 13 (S)-4-oxo-5-(jert-butoxycarbonylamine/)-6-(4 -benzyloxyphenyl) Production of bovine acid (Boc-(0-benzyl)tyrosi) N-N-methoxy/-N-methyla upper left a) Boc-(0-benzyl)tyron 7.42 in 20 ml of dry CH,CI 1 volume of g (30,0 mmol); at night, add 3°0 ml of triethylamine ( 21,5 mmol) and benzotriazol-1-yloxytris (dimethyl Ruami/) Phosphonium hexafluorophosphate 6.96g (20.0ml) (remol) was added. After 10 minutes, add 3.0 ml of triethylamine (21.5 ml) rimole) and dimethylhydroxylamine 2.15 g (22.0 mmol) added. After 16 hours, the solution was diluted with ethyl acetate, 5% hydrochloric acid, saturated aqueous carbonate. Washed successively with sodium hydrogen and saturated aqueous sodium chloride. The organic solution The liquid was dried (MgSO,), filtered, and concentrated to a foam. /Hula on Rikagel chromatography (hexane: ethyl acetate (50: 50)), 5.87 g (yield 71%) of the title compound was obtained as a white powder.

NMRTCDC131: δ7.40 (5H, ml; 7.02. (4H) , dd; 1.38 (9H, sl.

MS: m/z (M+I()”415; 339.341.315 (6S)- 7-(4-benzyloxyphenyl)-6-(Lert-b) Argon atmosphere In 100 ml of anhydrous diethyl ether, 6.68 g of magnesium powder (0 , 275 mol) a few drops of 4-bromo-1-butene were added. Reflux Upon initiation, 11-bromo-1 is added at a rate such that a slow reflux is maintained. -2472 g (0,183 mol) of butene were added. After the addition is complete, the mixture is The mixture was stirred for an additional 30 minutes. In anhydrous THF, t-butyloxylponylthironne -(0-benzyl)-methoxymethylamide 18.95g (0,046 mol) A solution was prepared and added slowly to the Grignard reagent prepared above at 0°C. 9 After 0 minutes, TLC showed the reaction was complete and was quenched with 10% aqueous hydrochloric acid. reaction mixture The product was extracted three times with ether and the combined organic extract was dissolved in 1M hydrochloric acid and saturated aqueous bicarbonate. Washed successively with thorium and saturated aqueous sodium chloride. The solution was dried (MgS O,), filtered and evaporated under reduced pressure to give a white solid.

By chromatography on silica gel, eluting with 17% ethyl acetate/hexane. Purification to give 18.1 g (96%) of the title compound as a white solid.

NMR (CDC13): δ7.42 (5H, ml; 6.96 (4H, d d); 5.72 (LH, r+t); 5.10 (IH, br d)); 5 ．． 06 (2H, s); 4.95 (2H, m effect 4.4B (IH/(1); 2-95 (2HF m) 72.5-2.0 (4M 1ml; 1-38 ( 9H#S).

MS: m/z (M+H)'410.371,354.310 (5S)- 4-oxo-5-(tert-butoxycarbonylamino)-6-(4-bendi C) ozone at -78°C until the blue color persists. in CHCl.

8.0 g (19.6 mm molar) solution. The solution was then purged with argon for several minutes, followed by 6 ml of methyl sulfide was added. The resulting mixture was stirred at room temperature for 2 hours. , and then evaporated. Dissolve the oily residue in 1715 ml of acetone and cool to 0°C. did. Jones reagent (12 ml, 2M solution) was added dropwise and the mixture was diluted with isopropyl. Stir for 10 minutes before quenching with the tool. Pour the solution underwater and add dichloromethane. Extracted with Tan three times. The combined organic extracts were dried (\,1g5O,) and filtered. and evaporated under reduced pressure. The oil was first mixed with ethyl acetate:hexane (2: 3) and elute with acetic acid: ethyl acetate, hexane (5: 35: 60). Purified by column chromatography on silica gel as a light brown solid. 3.39 g (yield 40%) of the title compound was obtained.

NMRfcDc13): δ7.40 (5H, ml; 7.0 (4H, dd ); NMR (CD30D): δ5-O5 (IH/obsc, l; 5-03 (2HF sL4.52 (LH, elk I; 3.2-2.5 +7H, ml; 1.40 (9H, s).

Example 14 (3R,4S)-4-(N-benzyloxycarbonyl)ami/-2,2-diph Filling of fluoro-3-hydroxy/-5-phenylpentanoic acid (3R,4S)-4-(N--bennoloxycarbonyl)ami/-2,2-di Fluoro-3-human Ox/-5-phenylpentanoic acid a) Dry Methyl chloride 1710 ml, 0.77 g of N-Cbz-L-7 enylalanol (2.75 mmol ) was desiccated in tSml of dry methylene chloride in a slow steady flow. Dess-Martin periodinane 1.28 g (3.02 mmol) of the solution was added. After stirring for 30 minutes, 1 A solution of 1 part 0% aqueous sodium thiosulfate and 1 part saturated aqueous sodium bicarbonate 100 ml was added, the mixture was extracted three times with methylene chloride, and the combined organic extracts were Dry over magnesium sulfate/rum, filter and concentrate under vacuum to give N as a yellow solid. -Cbz-L-phenylalanal was obtained. Zinc 0.5 in 15 ml dry THF 4 g (8.24 mmol) and 60 mg (0°22 mmol) of CuBr. Suspension i& was added to 6.04 ml of diethylaluminium chloride (6.04 mmol, IM/hexane) and cooled to -20°C. This suspension was added with dry Tl. - In 25ml of lF, crude N　Cbz　L-phenylalanal and promo A solution of ethyl nofluoroacetate was added dropwise over a period of 20 minutes. The reaction mixture was ．． Stirred for 5 minutes, then quenched by adding 5% aqueous hydrochloric acid and warmed to room temperature. Applicable Extract the kelp three times with diethyl ether, and add the combined organic extract 1α with magnesium sulfate. dried over vacuum, filtered and concentrated. Flash chromatography on silica gel diastereomer (diethyl ether:hexane (1,5:1)) 0.78 g (70%) of a 3:2 mixture of The diastereomer is repeatedly fluorinated. chromatography (hexane:diethyl ether (1:1)) Separation gave 468 mg (42%) of the title product as a white solid.

3.01 (2H, ml; 1.31 (IH, t, J = 7.5 Hz + b) 1.0 g (2.45 mmol) of the ester of Example 14(a) was added to methanol:T It was dissolved in 40 ml of HE (3.2). The solution was dissolved in 25 ml of water with 2.5 mL of potassium carbonate. 7 g (19.6 mmol) and the mixture was stirred vigorously for 3 hours. The water soluble The solution was washed with diethyl ether, acidified with hydrochloric acid, and washed once with diethyl ether. Extracted twice with methylene chloride. The combined organic extracts were dried with magnesium sulfate. Filtered. The solvent was evaporated under vacuum to give 0.93 g (9 0%) was obtained.

NMR (CDC13): δ7.21 (1OH, ml; 4.91 (3H, m l; 4.22 (3H, ml; 2°97(2) 1.m) MS: m/z 380 0.35 g (1.0 mmol) of the compound of Example 2(d) was added to 50% methylene chloride. DCCo, 2’l g (1,0 mmol) and HOBTO, Val-Vat-B[1 (IO mmol) was added overnight using 5 g (1 mmol) of Val-Vat-B[1 (IO mmol). mole). Quantitative ninhydrin test showed 83% ligation. ninhydrin The peptide resin was treated with ActO (lml/methylene chloride 3) until the test was negative. 0ml) and acetylated with r:.

The peptide was then subjected to FA using half of the resin according to the procedure of Example 1. and acetylated. The peptide-resin intermediate was anhydrous for 1 hour at 0°C. Cleavage was performed using 0 ml of HF1 and 1 ml of anisole. The peptide was treated with glacial acetic acid. Extracted from fat. The crude peptide (37%) was obtained by lyophilization. Direct the peptide Purified by flow distribution method (n-butanol:acetic acid:water (4:1:5)) 60 mg was obtained. The peptide was further prepared by HPLC, Haffliton PRP-1305x7mm semi-prepared HPL C column, water-acetonitrile-○, 1% TFA (95:5-78:22) /13 minutes, (78:22-60.40) /6.5 minutes) to obtain 179 mg. Ta.

TLC:R,0,34(B:A:W(4:l:l),0.68(B:E :A:W(1:l:l:l)); HPLC: ("Milton PRP-1250x4.1mrn analytical HPLC column , water-acetonitrile-〇, 1% TFA (9 minutes, 5-60:40/15 minutes), k' 2.73: FAB-MS: m/z 817 (M+H)' ; Amino acid analysis: Ser O, 57, Gln boo, Asn 1.00 ゜Val　　2.00 Same as above except using 1.1 molar equivalents of butyryl chloride instead of acetic anhydride. 2-(Petyllucerylglutaminyl asparaginyl)amino-3-phenylene Lupropyl-brolylvalylvarinia (2-serylglutaminyl asparagini) ) Production of ami/-3-phenylpropyl-prolyl valyl valinamide Similarly, the title compound 8. 1 Omg was obtained.

R,0,93(B:A:W(4:1:’l),0.97(B:E:A:W (1:1:1:l)): HPLC: (Hamilton PRP-1250x4.1mm analysis HP L C cap) Rum, water-acetonitrile-o, i% TFA (95,: 5-60, 40/15 min), k’ 2.87; FAB-MS: m/z 775 (M )"Example 17 4-(acetylucerylglutaminyl asparaginyl)amino-3■ヱS)-H Droxy-1-oxo-5-phenylpentyl-pro1//L, Valylvaline Preparation of amide 103 mg of the title compound was obtained in the same manner as above.

TLC: (B:E:A:W(1:l:l:1))R1=0.51;F AB-MS: m/z 875 (M+H)”; Amino acid analysis: Asp= 1.00, 5et=0.14, Glu=0.94, Pro=1.09, Val= 2.24; Peptide content (based on Asp) = 80.8% 4-(acetyl-cerylglutaminyl-asparaginyl)-amino-3(R5)-h Production of droxy-1-oxo-5-phenylbenzirut sl rylvalinamide 21 mg of the title compound was obtained in the same manner as above.

TLC: (B:E:A:W(1:1:l:1))Rr=0.33 ;HPLC: (4.5mmx25cm Altex Ultrasphere -(Altex Ultrasphere) 5m ODS, acetonitrile Gray water - 1% TFA, acetonitrile 10-50% gradient, 20 minutes time), k’ = 4.47.4.37; FAB-MS + m/z 778 (MH)'; Amino acid analysis: Asp=1.0O1Set=0.71. GIu= 1.00, Val=2.09; Peptide content (based on Asp) = 72.75% Daiarashi↓bi 4-(serylglutaminylasparaginyl)amino-3(R) Same as above. 20 mg of the tM peptide was obtained.

HPLC: (4.6mmx25cm Altex Ultras, Fair -〇DS, 15% acetonitrile-water-0.1% TFA) k' = 6.33; FAB-MS: m/Z 736 (M+H)'; Amino acid analysis: Asp= l, O05Ser=0.71, Glu=0.99.4-(seryl glutamini rusparaginyl)amino-3(S)-hydroquine-1-oxo-5-phenyl Production of pentyl-valyl/<phosphorus amide 12 mg of the title compound was obtained in the same manner as above.

HPLC: (4.6mm x 25cm Altex Ultrasuff Air -〇DS, 15% acetonitrile-water-0.1% TFA) k' = 633: FAB-MS: m/z 736 (M+H)'; Amino acid analysis: Asp-1 ．． 00.5er=0.7L Glu=0.99, Val-1,96 Example 21 (IR,2R)-2-(((2S)-2(tert-butylox7carbonylcetate) lylalanylalanyl)amino-1-oxo-3-phenyl)furovir)socro Production of Bentylcarbonylruvalylvaline methyl ester (IR,2R)-2(((2S)-2-tert-butyloxycarbonylamine (1-oxo-3-phenyl)propyl)cyclobentyl sulfonyl Luvaline methyl ester (a) THF (0,4r A solution of the compound io of Example 3(f), smg (29,0 mmol) in nl) and N-methylmorpholine 6rr+I (55 mmol) and isobutylene chloroformate. 3.8 ml (29.1 mmol) of Tyril was added. After 20 minutes, another N-mail Add 6 ml of tilmorpholine, add 6 ml of tilmorpholine, and dissolve in THF (0.5 ml). A solution of 16 mg (60 mmol) of methyl ester hydrochloride was added. the mixture The mixture was stirred at -40°C for 30 minutes and then at 20'C for 17 hours. Add the mixture to vinegar Diluted with ethyl acid, washed with 5% hydrochloric acid, dried over magnesium sulfate, and! ! Shrunk. Hula.

'ixri07) Graph 4-(CHCI 3: CH301 ((:)O: l) ):: 13.4 mg of the title compound as a white solid (23 mmol, yield 8 1%).

TLC: (CHCI3:CH,OH(50:1))R1=0.28:N MR (CDC131: δ7.26 (3H, m; ffl, tl), 7. 10+2H, m, 14fil.

6.50 (IH, d; J = 8.7 Hz; NH), 6.35 (IH, d; J-8,6Hz; NH), 5. LL (IH, d; J-7, 91Az; BocNH), 4.70+13(, itl et tert hammer, j1$le l/ 4.52TIH, dd; J-8,7,4,9Hz>, 4. 26 (IH, dd; J-8,6,7,1Hz+, 3.73 (3H, s), 3.51 (IH, IIF et o' to the east of the prefecture 1. 3. 16 (1M, dd,; J-14, 1゜0.941-0.874 ( 12Hr + 2 tic.

MS (FAB+VE): m/z 574 (M+H)", 518.474. 443.387.343.244.229(IR,2R)-2-(((2S)- 2-amino-1-oxo-3-phenyl)propyl)/clopentylcarbonyl -valylvaline methyl ester, trifluoroacetate (b) of Example 21(a) 13 mg (23 mmol) of the compound was dissolved in 0.5 ml of trifluoroacetic acid. 2 After an hour, the 1 g solution was concentrated under vacuum, and the residue was combined with methylene chloride and concentrated again. , 13 mg of the title compound was obtained as a white solid.

(IR, 2R) 2-(((2S)72-(tert-butyloxycal Bonyl=(0-phenylmethyl)serylalanyl-alanyl)ami7-1-oxy So-3-phenyl)propyl)cyclopentylcarbonyl-valylvaline methyl Ester (C) Boa-3er (Bzl)-Al at -40°C under argon a - Add N-methylmorpho to a solution of 15.1 mg (34.5 mmol) of Ala. 7.7 ml of phosphorus and 4.5 ml of isobutyl chloroformate (35 mmol) Added. After stirring for 20 minutes, 77 ml of N-methylmorpholine was added to the colander. Subsequently, a solution of 13 mg of the tripeptide of Example 21(b) in THF was added.

The mixture was warmed to 20°C and stirred overnight. Dilute the reaction mixture with methylene chloride and washed with 5% hydrochloric acid, 5% NaHCO, and water. Crude raw material is produced by evaporating the solvent. The product was obtained and subjected to furanone chromatography (gradient elution, CHCl =:cHsOH(50:1-25:1))l: marked as a white solid 17.9 mg (20 mmol) of product was obtained.

TLC: (CHCl, :CH30H (95: 5)) R+ = 0.35; ( 3H, sl, 3.78-3.62 (2H, m: BnOC! 121.3.4 0 (IH, ml, 3.20 (LH.

dd; 3.09 +LH, m), 2.92 +IH, ddG J -14, 4, El, 7 Hz; n: +9v), 2.25-1.95 (4 H, ml, 1.85-1.65 (4H.

ml, 1.47 +98. , s), 1.35 (3H, d; J = 7.2 Hz), 1.23 (3H, d; J-7, 2Hz), 0.956-0.922 (12H, !L, ?It two skins).

MS (FAB + VE): mlZ 893 (M + H) +, 793 ．． 762.706.662.645°634, 514.474.420.364 ゜(IR, 2R) -2-(((2S) -2-(tert-butyl oxy7cal Bonylserylalanylalanyl)ami/-1-oxo-3-phenyl)propyl ) cyclobentylcarbonylvalylvaline methyl ester (d) 1 mg of hexapeptide 8 of Example 21(c) in 4 ml of methanol ( A solution of 10% Pd/C (7 mmol) was prepared under hydrogen atmosphere for 15 h. g) and stirred together. Filter and evaporate to give 7.2m of the title compound as a white solid. g (9 mmol) was obtained.

TLC: (CHCI=: c) (3o) ((95: 5)) Rr=0,10 : Example 22 (IR,2R)-2-(((2S)-2-(serylalanylalanyl)amino- 1-oxo-3-phenyl)propyl)cyclopentylcarbonyl-valylvari Preparation of methyl ester, hydrochloride 5 mg of the compound of Example 21(d) (6.2 mg) rimole) was dissolved in trifluoroacetic acid (0.25 ml). After 90 minutes, the solution Concentrate under vacuum, dissolve the remaining IM in methanol (0.5 ml) and add concentrated hydrochloric acid (approx. ．． 025ml) was added, the solution was concentrated, and the resulting gum was triturated with ether. Turate and dry under vacuum to obtain 4.5 mg of the title compound as a white solid. Ta.

NMR (CD30D): δ7.20 (5H, ml, 4.72 (LH, d d; J-8,5,5,0)1z).

4.38 (IH, ml, 4.31 (2H, ml, 4.19 (LH, d ), 3.92 (3H, ml, 3.70 (3H, s), 3.48 (LH , ml, 3.19 (IH, dd; J-14, 3, 5, 0 Hz; <=vl a L 3-O8 (1) Hz m), 2-83 IIH, dd; J-14-3t B-5Hz; tri 21 Itogame)・ MS (FAB + VE): mlZ 703 (M + Ml”.

Example 23 (is, 2S) 2 (((IS, 2S) 2-(jerk-butyl oxy/carbonyl-serylalanylalanyl)amino-1-hydroxy-3-ph (enyl)propyl)cyclopentylcarbonyl-valylvaline methyl ester manufacturing (Is, 2S) 2 (((IH3,2S)-2tert-butyloxy Cyclobonylamino-1-hydroxy/-3-phenyl)propyl)cycloben Kunryurubonyl-L-valyl-L-valine methyl ester a) 35.5 mg (97.8 mmol) of the product of Example 8 and HOBT26 rng (196 mmol) in dioxane Q, 5 ml and DMFQ, 1 mmol. Dissolve and add 20.2 mg (98.1 mmol) of DCC. Stir for 20 minutes After that, 32 ml (290 mmol) of N-methylmorpholine and Valylvari 52 mg (195 mmol) of methyl ester hydrochloride were added. After 3 days, the mixture The material was diluted with ethyl acetate, filtered, and extracted with dilute hydrochloric acid. The organic layer was concentrated. The coarse The product was purified by flash chromatography (CHCI, :CH,OH (50:l )), the first eluted isomer A 26 mg <45.9 mmol, yield 47 %) and 825 mg of the isomer (43.1 mmol, yield 44%) were obtained.

Isomer A; (Is, 2S) 2 (((IR, 2S) 2 (tert-butyl half-dicarbonyl)amino-1-hydroxy/-3-phenyl)propyl)/klope Tanthane carbonyl valyl valine methyl ester TLC: (CHCl=: CH, OH (25: 1)) R, = 0.55 + NMR (CDC131: δ7. 28 (5H, ml, 6.73 (IH, d; J”8.7 Hz; NHl, 6 -42 (IHt d; J-8,6Hz; NH)l 5.16 (IH/d; JW9-5 Hz; BocNHl.

4.58+IH, dd; J”8.8.5.2 Hz), 4.32 TIH, dd; J-8,6,6,0Hz).

3.91 (IH, light 5b, rb at+t(4), 3.76 (3H, s) e 3.25 ('y, ml et ts+r, f-21411 (; J-10, 2Hzl, 2.95-2.83 (2H; tr-Shi-I-Ra-1, 2, 5 2 (LM.

*Ara,, rl l-@@), 2.30-2.10 (38, ml, 2.00-1.55 (6H, mL Isomer B: (Is, 2S) 2 (((is, 2S) 2 (teri-butyl (oxycarbonyl)amino-1-hydroxy-3-phenyl)propyl)cyclope TLC: (CHCI3・C H30H (25: 1)) RI = 0.45: NMR (CDC13): δ7.2 4-7.15+5H, mL 6.77 (IH-d; J-8, 3Hz), 6-5 6(l)l, d; J-8, 2Hz), 4-80(IH, d; J-8 , 4Hz) t '4.47 (IH, dd; J-8, 4, 4, 9Hz), 4 ．． 36 (IH, clear nIrJ mit4>, 3.5o-3,6o+sH , ml.

3.08 (IH, dd)l 2. '69 (2H, mi,?;u'l ,)1)/2.5] IH, ml, 2.25-2.05+28.　 ml, 2.00-1.60 (6H, m), :L, 32 (9H, s), 0 ．． 979-0.855 (12H,! 1 ball from Sha).

MS (rAB + VE): m/z 576 (M + H)", 476, 389.246.231.152.132゜(], S, 2S)-2-(((is , 2S)-2-amino-1-hydrocow/-3-FJ,nyl)propyl)cyclobe Tantancarponyluvalylvaline methyl ester, tgui b) 825 mg (43 mmol) of the isomer of Example 23 (a) was converted into trifluoro Dissolved in 0.25ml of acetic acid. After 1 hour, the solution was evaporated on a rotary evaporator. Concentrated. Dissolve the shallow pickles in CH, OH, react with 0.05ml of concentrated hydrochloric acid, and re-incubate. It shrunk. / trituration with ethyl ether, dried under vacuum, white color 22 mg (43 mmol) of the compound described above was obtained as a powder.

(Is,2S)-2-(((IR,2S)-2-ami/-1-hydroxy-3- phenyl)propyl)cyclopentanecarbonyl-valylvaline methyl ester , hydrochloride C) 26 mg (45 mmol) of isomer A of Example 23(a) was added to Example 23 Following procedure (b), the title compound 19 was obtained by treatment with trifluoroacetic acid and hydrochloric acid. ．． 9 mg (39 mmol) was obtained.

(l S, 2S) 2 (((ls, 2s)-2-(tert-butyloxy cycarbonyl-L-(0-phenylmethyl)serylalanyl7ranyl)amino- 1-Hydroxy(3-phenyl)propylenecitalobentylcarbonylruvalyl valine methyl ester d) B in 3 ml of THFo at -40 °C under argon. oc-5et(Bzl)-Ala-A1a 19.4 mg (44.4 mmol) to a solution of 8.8 ml of N-methylmorpholine and 5.8 ml of isobutyl diroformate. ml (44.4 mmol) was added. After stirring for 20 minutes, roughly Add 8.8 ml of lumorpholine, followed by THF 1. In Qml, Example 23 A solution of 22 mg (43 mmol) of compound (b) was added. The mixture was heated to 20°C. and stirred overnight. The reaction mixture was diluted with methylene chloride and washed with 5% hydrochloric acid. Cleaned, dried (magnesium sulfate) and concentrated. Treat the residue with furanone chromatog 25 mg (28 mmol, yield 6) of the title compound as a white solid. 5%).

NMR (CDC13): δ7.40-7.0'5 (11H, ml, 7.0 0-6.80 (4H, m; NH); 5.53 (IH, d; BocNH) , 4.52 (2H, s), 4.47-4.30 (4H, ml, 4.19-4.061.60 (4H, ml, 146 T9H, s ), 1.32 (3H, d; J-7,0Hzl, 1.15 ( 3HB 664, 618.565.547.476, 416.388° (13,2S)2 (((IR,2S) 2 (tert-butyloxycarbonyl-L- (0-phenylmethyl)serylalanylalanyl)amino-1-hydroquine-3 -phenyl)propyl)cyclopentylcarbonyl-valylvaline methyl ester e) Using 19°9 mg (39 mmol) of the tripeptide of Example 25 (C) , In the same manner as above, 23°2 mg (26 mmol, A yield of 67% was obtained.

NMR, (CDC13): δ7.38-7.17 (10H, lT11.7. 10 (2H, m; NHI, 6.96 (IH.

Moerari, rr 4ll shudder L 3.84 nu, dd; J-9,4 , 4, 6H2I, 3.74 (3H.

(15,25)-2-(((is,2S)-2-(tert-butyloxycarp) (bonyl-1-serylalanylalanyl)amino-1-hydroxy-3-phenyl ) Furovir) Cyclopentylcarbonyl-valylvaline methyl ester f) 23 mg (28 mmol) of the compound of Example 23(d) in 1 ml methanol ) with 10% Pct/C (12 mg) under hydrogen atmosphere for 15 hours. Stirred. Filter and evaporate the solvent to obtain 20.7 mg of the title compound as a white solid. (25.7 mmol, yield 92%) was obtained.

TLC: (CHC1,:CH,OH(95:5))Rr=0.39:N MR (CDC13): δ7.71 (IM, 7-roto-; NH ), 7.58 (l)! ,　　F...low)”　　пG　NH).

7.16 (5H, ml,, 6.97 (2H, m; NH)/6. 12 TIM, >o-)”;NH)r 4-75(iH,>ch-?’:; BocNH), 4.42 (iH(, dali, 4.40-4.04 (S R, ml, 3.89 ・(l)! , 7・o-1”), 3.70 (3H , 5), 3.64 +2H, ml, 3.03 (IH, dd), 2.82 -2.44 (4H, ml, 2.23-1.87 (44ml, 1.80- 1.60 (4B, m), 1.45 (9H.

s), 1.35 (3H, d; J-6,8Hz), 1.09 (3H, d ; J-6,9Hz), 0.963 (6H; 2) / 1! practice), 0.8 53 (6H, nihu ni l yabaku).

MS (rAB + VE): m/z sos (M +)ri”, 789 ．． 705.674.618.574゜476, 416.388.300゜ MS (E'AB-VEI: m/z 803 (M-H)''.

Example 24 (Is, 2S)-2-(((Is, 2S)-2-(L-seryl-shi-alanyl- (alanyl)amino-1-hydroxy-3-phenyl)propyl)cyclopene Production of tylcarbonyl-valylvaline methyl ester, trifluoroacetate 1.3rr+g (1.6 mmol) of the product of Example 23(f) was treated with trifluorofluoride. Dissolved in acetic acid. After 90 minutes, the solution was concentrated to dryness under vacuum. Ete the residue The ether was removed in vacuo to give the title product as a white solid. A xapeptide was obtained.

Example 25 (IS, 2S) 2-(((IR, 2S)-2(tert- 1-hydroquine-3-phenyl) propyl)cyclopentylcarbonyl-valyl-L-valine methyl ester Manufacturing of According to the operation in Example 23 (f), the hexape of real t=, l'1J23 (e) 23 mg (26 mmol) of peptide was debennorized to form the title compound as a white solid. 18.9 mg (23.5 mmol, yield 90%) of 7 mg TLC: ( CHC1,:C)1. OH (93: 5)') R, = 0.43: N calm (CDC 13): δ7.47 (L Town dj J Rin 7.3 Hz: NIH), 7.3 4 (IH, d; j ring 6.8Hz: NH), 7.29-7.16 (5 H, ml, 7.09 (IH, d; J-8,9Hz, NH), 6.79 (LH, d; J"8.5 Hz; NH), 5.79 (LH, d; J-6,5Hz; NH), 4.76 +1 formula% formula% Production of roacetate In the same manner as above, the title compound was obtained as a white solid.

Example 27 (IR,2Rn-2-((IS,25)-2-(tert-)'tenop i-oxycarbonylserylalanylalanyl)ami/-1-nitroxy-3-phene Production of cyclopentylcarbonyl-propyl-valylvaline methyl ester Construction (IR,2R)-2-(((IH5,2S)-1-human:xy-2-tert- butyloxycarbonylamino-3-phenyl)propyl)cyclopentylcal Bonyl-valylvaline a) Epimer of Example 5 38m in 4 ml of dioxane Q g (0°10 mmol), add 27 m of N-hydroquine benzotrimazole g (0.20 mmol) and DCC22 mg (0.i 1 mmol) were added. I got it. After stirring for 20 minutes, 27 ml of N-methylmorpholine (0°25 mmol) ), followed by 33 mg of barylvaline methylnistil hydrochloride (0.20 mg). Rimol) was added. The mixture was stirred for 5 days and then ? Filtered and concentrated.

Attached to furanosinichromatography (CHCIs/CH=OH (20:1)) and white; g (0,08 I got 9mm.

TLC: (CHCI,:CH,OH(20:1))RI=0.37: 2.95-2.58 (3M, m), 2.43-1.55 (8H, ml, 1 ．． 45-1.25 (9H, tw.

(IR,2R)-2-(((IH5,2S)-1-hydroxy-2-amino-3 -phenyl)propyl)cyclopentylcarbonyl-valylvaline methyl ester Le, 1ai b) 48 mg (83 mmol) of the peptide of Example 27 (a) was treated with trifluoro Dissolve in 5 ml of acetic acid Q.72. After 1 hour, the solution was evaporated on a rotary evaporator. Concentrate, dissolve the residue in CH30H, react with 1 ml of concentrated hydrochloric acid Q, and condense again. 72. Tritinilated with ether and dried under vacuum as a white powder ( 42 mg (82 mmol) of Hyo Ki compound was obtained.

(IR,2R)-2-(((IH5,2S) 2 cycarbonyl-(○-phenylmethyl)-serylalanyl7ranyl)amino-1 -Hydroxy(3-7enyl)propyl)cyclopentylcarbonyl-valilva Phosphorus methyl ester C) Bo in 4 ml of THFo at -40°C under argon Solution of 35 mg (0,080 mmol) of c-Ser(Bzl)-Ala-A1a 11 ml of N-methylmorpholine (0.10 mmol) and chloroformin were added to the solution. 10.4 ml (0,080 mmol) of isobutyl acid was added. Stir for 20 minutes After that, 11 ml of N-methylmorpholine was added, and then the solution was added in 1 ml of THF. A solution of 42 mg (0,082 mmol) of the compound of Example 27(b) was added. Applicable The mixture was stirred overnight while warming to 20"C, then dissolved in methylene chloride and diluted with Washed with % hydrochloric acid, 5% NaHCOy and water, the solvent was evaporated and the crude product 61 mg was obtained.

50 mg of the crude product was subjected to Furanosini chromatography (grasheft elution). , CHClt: CH30H (Do: l~25~i)), 6 mg of pure isomeric Al, followed by 11 mg of the mixed fraction, and finally pure 3 mg of isomer B was eluted 7:.

Isomer B (IR, 2R)-2(((Is, 2S) 2 (tert-butyloxyca Rubonyl-L-(0-funinylmethyl)serylalanylalanyl)amino-1- Hydroxy-3-phenyl)propylenecyclopentylcarbonylvalylvari Methyl ester TLC: (CHC], :CH30H(25:1))R , = 0.40: NMR (CDC13): δ7.40-6.95 (15H, m; 7'llL + NH), 5.58 (IH, clBocNH), 4.53 (21,s; 蓼=Jizuri, 4.51(lLdd), 4.37T2H, m), 4.26-4.20 (3H, m), 3.7 5 (2H, m), 3.71 (3L s), 3.54 (IM, 7”O-)’lz2.96-2.90 (2M; He-’,::rvli>, 2.69+IH, ml, 2.38 (LH, ml, 2.15 (2H, m; CM(CH3)2), 1.90-1.60 (6H, mL 1.46 (9H, S:　Boc).

1.37 (3H, ei; J-7, 2Hz; CH3), 1.20 (3H, d; J-7, 2Hz; CH3), 0. X37 -0.980 +12L tg 2 la.; CH3).

Isomer A (IR, 2R) 2 2” (((IR, 2S) 2” (tert-print) tyloquine, carbonyl-L-(○-phenylmethyl)serylalanylalanyl) Amino-1-hydroxy-3-phenyl)propyl)cyclobentyl -valylvaline mether ester TLC: (CHCl,:CH,CH(20: 1)) Rr=0.40: NMR (CDC13): δ7.79 +14H , d; NO), 1.40-1.10 (11)! , rn; 7 ・)n:@+ N1(), 6-10 (2H; NH)t 5-59 <IH2d; J-3-3Hz; N13)t 4-93 (IH2пG *-3,2Hz)t　4.5<　(2!(/　　　S;　Be>!-11.-+ E, ), 4.4.7 (IH, CMd; , 7 | 8.5° 5.0 Hz), 4.30-4.20 (4H, ml, 4.14 +1 formula m+, 3.78 +IL ml, 3.7■ (3Htg), 3.61(IHtml, 2-90CIHt(icl;, Y=C tkd"-Jt2-1"7 (2H/rn)z2.35-2.10 44H, ml , 1.95-1.55 (6M, m), 1.49 (9)! , s; Boc), @ 1.40 (3H, d; J”7.3 Hz; CH3), 1.12 (3H , d; J-7,3Hz; CH3)t o-99(64”’2’lt r! c; CH3)z O, 91 (6Ht = tr:'-1+tk ; CH3).

(IR,2R)-2-(((Is,2S)-2-(tert-butyloxycal) Bonylserylalanylalanyl)amino-1-hydroxy-3-phenyl)pro pill) cyclobentyl luponyl (lyl i < phosphorus methyl ester d) 813 mg (15 mmol) of the isomer of Example 27(c) was subjected to the above procedure. Therefore, 12 mg (15 mmol) of the peptide was obtained as a white solid.

TLC: (CHCl, :CH, CH (95: 5)) Rr = 0.21; N MR (CD30D): δ1.21 (sg, m), 4-35 +1)! 7m) , 4-30-4.13 (49m).

4.08 (E, >o-F),' 3.85 (IH, dd; J -10,8,5,4F,z), C320H), 3. V3 F1B, m), 3.72 (34s), 3.58 +1L a t et al. 1, T μ cut t) C4, 3dZ) I 2.112 (IH, dd; , 7-13.7. 6.2 Hz; /? =T, :+5)q), 2 ．． 75　(YuH,ddy　　e＞Nshi噤{l+.

2.71 (11, mH, 2,37 (IL″7-o-dom), 2.19- 2.02 (2H, m; C-1(CH3)2>, 1.85-1.60 (6H , ml, 1-48 (9Hr s; Boc), 1-41' (3) i- d; 274.246, 228, 203° Example 28 (I　R,2R)-2-(((I　S,2S)-2-(serylalanylalanyl )amino-1-hydroquine-3-phenyl)propyl)cyclopentylcarbonyl Production of rubalylvaline methyl ester, hydrochloride Using 12 mg (15 mmol) of the product of Example 27(d), the same procedure as above was carried out. 11 mg of the title compound was obtained as a white powder.

NMR (CD30 (1!l: δ7.23 (5H,, m), 4.50-4. 2513H, ml.

2.32 (1M, m), 2.20-1.95 (2H, m), 1. E.I. O-1,58 (6H, ml.

1.40 (3H/d; J-7,1K!1/1.27 (3H t d: J-7, 2Hzl.

0.959-0.931 (12H, 21 Fukotsuchi).

MS (FAB + VE): m/r: 705 (M 1 H)”, 574. 476° Example 29 (IR,2R)-2-(((IR,2S)-2(terL-bufAtxycarboxylate) Nyl-serylalanylalanyl)amino-1-hydrocythyl-3-fninyl)pro Production of cyclopentylcarbonyl-valylvaline methyl ester Example 27 Isomer A1.5 of (C). ．． 12 using 5 mg and following the procedure above. 13.8 mg (17,: 2 mmol) of peptide was obtained.

TLC: (CHC1,:cH,OH(95:5))R,=0.27:C f120H), 3.72 (3H, sL 3.5B (IH, dd; J-10,0,),,8),2.98+IH,ddG 5m14. 1, 3.4 Hz; 2.71 (2H1ITI), 2.34 (LH121 bar-; C1(f c! 13)2).

MS (E”8B + VE): m/z 805 (M + H)", 674. 618. 574. 519. 4V5゜ 308.264° Example 30 (IR,2R)-2-(((lR,2s)-2-(serylalanyl-shi-alani) (l)ami/-hydroxy-3-phenyl)propyl)cyclobentylcarbonyl -Production of Barylha; Inmether ester, Hydrochloric acid; Using 13.8 mg of the hexapeptide of Example 29, the procedure of Example 28 was followed. Thus, 13 mg of the title compound was obtained.

NMQ (CD30D): 37.20 (5H, m), 4.39-4.17 +4H, ml, 4.07-3.92 C4H.

m), 3.70 (3H, s), 3.52 (IH, dd; J-9, 5, 2.9Hz), 2.98 fIH, dd; Ja*14.2. 3.4 Hz ; ff>u+4%), 2.82-2.68 (2H, mL 2 ．． 3B (IH/High k 2.21-1.55 (8H, ml, 1.35 (3H, d; J=7.2 Hz), 1.20 (3H, d; J -7,1Hzl, 1.01-0 ．． 889 (12H, @ff1I!I=qlf&,>.

KS (TAB + VT:): m/z 705 (M + Ml”, 575.476° Example 31 (l　R,2R)-2-(((IH3,2S)-2-(acetylserylalanyl alanyl)amino-1-hydroxy-3-phenyl)propyl)cyclopentyl Production of carbonyl-valylvaline methyl ester (IR,2R)-2-(((IH5,2S)-2-(acetyl=(O-phenyl) methyl)serylalanylalanyl)amino-1-hydroquine-3-funynyl)propylene ropyru) /7c pentylcarbonyl-valyl, NH zolyntyl ester a) Epimer mixture (i:1) of the compound of Example 27 (c) 10°8 mg ( 12.2 mmol) and following the procedure described above, the title epimer mixture was obtained as a white solid. Compound 9-2 mg (11.0 mmol) was obtained.

TLC: (CHCI=:CHsOH(20' 1)) Rt=O,'2v, O1N (CD30D): δ7.38-7.11 (10H, m), 4.5 7 (2H, s), 4.53-3.97 (6H, ml, 3.78 (2H, -e, w+r4 = @稗), 3.70 (3M, s), 3.68 (I H.

m), 3.53 (1) i, m), 3.00-2.59 (2H, ml, 2.35 (IH, rr+l, 2.18-1.98 (IR, 2R) -2-(( (IH5,2S)-2-(7cetylucerylalanylalanyl Propyl) cyclopentylcarbonyl-valylvaline methyl ester b) The above implementation Example 31 (a) shrimp? -9.2 mg (11.0 mmol) was used in the above procedure. Therefore, 8.2 mg of the title bimer mixture as a white solid (1'=, O mmol ) was obtained.

TLC: (CHC1,: CH, OH (9: 1)) Rt=0.35.0° 27: NMR (CD30D): δ1-21 (5H, m), 4-42-4-09 <6 x, m>r 3-89J-60 (2h-m), 3.70 (3H, s), 3.04 (LH, m), 2.99-2.59 (2H, ml, 2.34 (IH.

m), 2.18-1.99 (5H, ml, 1.92-1.55 (6B, m), 1.38 (3H,.

Example 32 trans-2-((1-methoxy-1-(2-phenyl-1-(tert-butene) Luozidicarbonylruserylalanylalanyl)amino)ethyl)phosphinyl ) Production of cyclopentylcarbonyl-valylvaline methyl ester trans-2-((1-methox7-1-(2-phenyl-1-phenylmethocino) Carbonylamino)ethyl)phosphinyl)cyclopentylluponylrubaryl Valine methyl ester a) Using 43 mg of the compound of Example 11(d), the above procedure 2.Therefore, 6.5 mg (98.9 mmol) of the title compound was obtained as a five-color solid. . Yield 99%.

TLC: (CHCl, :CH,OH (9:1)) Rr=0.6 Nidoran Su-2-((1-methoxy-1-(1-amino-2-phenyl)ethyl)phos finyl) cyclopentylcarbonylruva! , °luvaline methyl ester b) Using 30 mg (46 mmol) of the compound of Run 171132(a), Following the procedure, 20 mg of the title compound (38 mmol, yield 84 %) was obtained.

NKR (CDC13): δ7.38-7. i8 (5M, ml, 4.51 (1M, ml, 4.30 (IH, ml.

3.90-3.68 (61, m), 3.44-3.03 (2H, m), 2.93-2; 48 (IH, mH.

2-25-167 IBHt m), 1.27 (IHz ml, 1.01-0 ．． 85 +13H, m).

trans-2-((1-methoxy-1-(2-phenyl-1-(tert-butylene) (0-phynylmethyl)serylalanylalanyl)amino ) Ether) Phosphinyl) Cyclobenyl Valyl Valine Methyl Stell C) Collagen compound 2 as glassy crystals according to the procedure of Example 27(c) 3.5 mg (24.9 mmol, yield 65%) was obtained.

TLC: (CHCI=: CHsOH (9: 1)) Rt=0.5; NM R (CDC13): δ7.39-7. X3 (log, m), 4.80-4 ．． 20 (8H, m), 3.90-3.60 (sx, rrJt 3.45 -2.10 (3H, mat 2-22-1-64 (8H, mat!, 50 -1.25 (12H, m), 1. YuO (3H, ml, 1.04-0.8 5 (13H, m).

trans-2-((1-methoxy-1-(2-phenyl-1-(tert-butylene) Ruokidicarbonylrucerylalanylalanyl)amino)ethyl)phosphinyl ) Cyclobenticarbonyl-valylvaline methyl ester d) 19.1 mg of the title compound as a white solid according to the procedure of Example 27(d) (22 mmol, yield $93%) was obtained.

NMR (CD30D): δ7.32-7.16 +5H), 4.75-4. 10 (6M), 3J7-3.66 (8H).

3.32-2.72 (3H), 2.20-1.62 (811,1,50- 1,28+12H), 1.13 (3H).

1.05-0.135 +13H).

MS (E”AB+■): 853 (M+H)+, 753° Example 3 3 trans-2-((1-hydrogen-1-(2-phenyl-1-(serylalani) Rualanyl)amino)ethyl)phosphinyl)cyclopentylcarbonyl-vari Production of Luvaline Methyl Ester Hydrobromide The above procedure was carried out using 9.6 mg (11.3 mmol) of the compound of Example 32(d). Accordingly, 9 mg of the title compound was obtained as a light yellow powder.

NMR (CD30D): δ7.30-7. i5 (5H, m), 4j5-3 ．． 92 <5H, m), 3.83-3.66+5)! , ml, 3.30- 2.08 (4H; m), 2.20-1.64 (84ml, 1.47- 1.25 (4H.

m), 1.12 (3H, ml, 1.05-0.90 +12H, ml.

MS (FAJI + VT:): mlZ 139t (M-H Bx + H) 10, 260. 232t L59, Example 34 (5S,4RS)-5(tert-butyloxycarbonylphurylalanyl ranyl)ami/-4-hydroxy-1-oxo-6-7enylhexylvalyl Production of valine methyl ester (3S) Ami/)-1,4-dioxo-6-phenylhexylvalylvaline methyl s a) Perform 1 month in 5 ml of THF at -40'C under argon. 2(b) = N-methylmorpholine to a solution of 160 mg (0,495 mmol) of 66 mg (0.66 mmol) followed by 68 mg of isobutyl formate (0,495 mmol) was added. The reaction mixture was stirred for 10 minutes and then 66 mg (0°66 mmol) of N-methylmorpholine was added, followed by 55 mg of THF. Valylvaline methyl ester hydrochloride 120 mg (0.45 mmol) in ml solution was added dropwise. The mixture was heated at -40'C for 30 minutes, then at 20'C for 1 hour. Stirred for 7 hours. Then, the reaction mixture was mixed with ethyl acetate: dichloromethane (1: 1) Dilute with 100ml and add 5% hydrochloric acid, 5% sodium bicarbonate and saturated chloride solution. ) Continuously washed with +/um aqueous solution I: 72. Dry the organic extract (magnesium sulfate) um), filtered and evaporated under reduced pressure to an oil. Flash on silica gel Subjected to chromatography (chloroform:methanol (25:1)), white 160 mg (yield 70%) of the title compound was obtained as a colored foam.

NMR (CDC13): 67.25 (54m), 6.34 (LH, d), 6.15 (IH, a), 5.24 (IPt, and).

4.58 (2M, m), 4.31 (LH, dd), 3.78 (3H, s), 3.19 (IH, m), 2.95 (LH, ml.

2.87 (21, m), 2.52 (2H, t), 2.21 (2 present five * Lt1 ．． 43<9xrs>tO+98 (12H, m).

MS (r13”): m/z 534; MS CF’AB-): m/ z 532 (5S, 4RS) 5-(tert-butyloxycarbonyl ) Amino-4-hydroie/-1-oquine-6-phinylhe beef silva+/rubari methyl ester b) Q'C+: 160 mg of the compound of Example 34 (a) in methanol (3, 0 mmol), 5.7 mg of sodium borohydride (1,5XIO-' mol) was added little by little. After 20 minutes, the mixture was diluted with 5% hydrochloric acid and dichloromethane. Extracted with methane three times. The organic extract was dried over magnesium sulfate, filtered and white Evaporated to colored flakes. Flash chromatography on silica gel ( Chloroporum S methanol (40:l)) and marked as white flakes 112 mg (70%) of the compound was obtained.

NKL (CDC13/MeODl: δ7.15+5H, m>; 4.38 ( IH, dd); 4.10 (1)! , m); 3.63 (3H, s); 3.0 to 1.5 (8M, m); 1.28+9H, s); 0.88 ( 12H, m).

(5S,4R3)-5-(tert-butyloxycarbonyl-(〇-phenyl) methylnserylalanyl-alanyl)amino-4-hydroxy-1-oxo-6 -Phenylhexylvalylvaline methyl ester C) 98 mg (0,183 mmol) of the peptide of Example 34 (b) was truffled. It was dissolved in 3mi of fluoroacetic acid and stirred at room temperature for 2 hours. Then, the solution was removed under reduced pressure. Concentrate it, dissolve it in methanol, add 3 drops of concentrated hydrochloric acid, evaporate under reduced pressure, and leave 86 mg of tripeptide amine was obtained in the form of colored flakes.

Boc-5er (O8z)-Al in THFSml at -40°C under argon. In a solution of a-Ala (87.4 mg, 0o20 mmol), N-methylmorpho was added. Add 25 ml of phosphorus (0.27 mmol), followed by isobutyl chloroformate. 27 mg (0.20 mmol) were added. The mixture was stirred for 15 minutes and then Add 25ml of N-methylmorpholine (0°27mmol), then add TH In 1 ml of F, previously prepared 5-amino-4-hydroxy-1-oxo-6-phini 86 mg (0.18 mmol) of nylhexylvalylvaline hydrochloride was added dropwise. . The mixture was stirred at -40°C for 30 minutes and then at 20°C for 17 hours. did. The reaction mixture was diluted with ethyl acetate:dichloromethane (i: ioom+). , 5% hydrochloric acid, 5% aqueous bicarbonate i-1) Rum, saturated aqueous sodium chloride continuously 2. Wash the solid thoroughly and remove the flash chromatography on silica gel until it becomes a white solid. Purified by chromatography (chloroform:meth/ol (20:1)), white 98 mg (yield: $63%) of the title hexapeptide was obtained as colored crystals.

NMR (CDC13/CD30D) 467.1 +5H, ml, 6.95 ( 5H, m), 3.48 (3H, 5L1-29 (9H, sat 1.0 5 (6H, m), 4.2-1.5 (mL 0-70 (12L r n).

Feather (3” on r): mlz 855; 85, (rJI-): mlZ 853(3S,4R3) 5-((tert-butoxycarbonyl-seryl alanylalanyl)amine)-4-hydroxy-1-τquin-6-phenylhexyl syl-valylvaline methyl ester d) in methanol:acetic acid (9:1), Example A solution of 31 mg (36.2 mmol) of compound 34(j) was added with 10% Pd/ 'Culture element (35 mg) was added. The mixture was stirred under hydrogen (l atm) for 6 hours. Then, the reaction mixture was filtered through Celite, and the Celite was mixed with acetic acid, meta/- and ethyl acetate. Evaporate the combined solution under reduced pressure. The title compound 21m (yield 75%) was obtained as white crystals.

NKR (CDC13/CD30D): δ7.14 (5H, ml, 4.33 (IH, ml, 4.08 (4H, mL3.60 (3H, S>, 2.95 (LH, dd), 2.70 (LH, m), 2.35 +11. ml, 2.06+24 ml, 1.72 (LH, ml, 1.40 (9L s) , 1.22 (6H, ri+; 0.85 (12H.

MS + TAB"): mlz 765; MS (FAB-): mlz 76 4゜Example 35 (4RS,3S)-5-(tert-butyloxycarponyl-SeInoralani) 6-(4-hydroxy) Production of phenyl)hexyl/slryl methyl ester (5S)-5-(tert-butyloxycarbonylamino)-1゜4-dio xo-6-(4-benzyloxyphenyllivaline methyl ester (a) Using 10 mg (1.9 mmol) of compound 8 of Example 13(c), In the same manner as above, 838 mg (769 mg) of the title compound was obtained.

(4RS,,5S)-5-(tert-butyloxycarbonyl)amino-4 -Hydroxy-l-oxo-6-(4-pendino-diphenyl)hexyl- Valylvaline methyl ester b) 144 mg of the compound of Example 35(a) (0. 223' mmol), and in the same manner as above, the title liberum was obtained as white crystals. 118 mg (yield: 82%) of butedo was obtained.

NMR (CDC13): δ7.4 +5H, ml, 7.02 (4H, dd), 6.40 (23f, rn), 5.03 (2H, s), 4.54 (2H, ml, 4.30 (LH, ml, 3 ．． 70 (3H, s), 3D61 +2H.

ml, 3.0 to 1.6 (11)! , ml, 1.38 (9H, s) , 0.91 (12):, ml.

(4RS, 5s) 5 (tert-butyloxycarbonyl) nylmethyl)serylalanylalanyl)ami/-4-heroquine-1-oxo- 6-(4-penzyloxyphenylenehexyl-valylvaline methyl ester C) Tripeptide 1 of Example 3.5(b) 10 mg (0.179 mmol) 40 mg of the title hexapeptide ( A yield of 25% was obtained.

N Associate (CDC13/CD30D+': δ7-35 (IOJ m), 6.91 nH, dci), '4.95 +2H.

d), 4.48 (2H, dl, 4.37 (17(, dl, 4.0 7 (4M,’ m), 3.62 <4F. , sr.

3, 60 (IH, obsc’m), 2’. 5 to 1. ! + (3H, mH, 1.38 (9Hz g), 1.2 (6':tm), 0.80 (12H,　m).

(4RS,,5S)-5-(Lert-butoxycarbo-ru-serylalanyl alanyl)amino)-4-hydroxy-1-oxo-6-(4-hydroxyphene) nyl)hexyl-valylvaline methyl ester d) Compound of Example 35 (c) Using 39 mg (40.6 mmol), following the hydrogenation procedure of Example 34 (d). Thus, 2.3 mg (yield 73%) of the compound described in item t was obtained as white crystals.

4, 2 to 3.8 (51 (, ml, 3.67 (3H, S>1 3 ．． O to 2.5 <2H, m>1 Example 36 (3R,4S)-4-(tert-buterotei dicarbonyl-seryl 7 ranila ranyl)-amino-2,2-difluoro-3-hydroxy/-1-oxo-5-ph Production of phenylpentyl-valylvaline methyl ester (3R,4S)-4-(benzyloxycarbonyl)amino-2゜2-difluoro rho-3-hydroquine-5-phenylpentanoyl-valylvaline methyl ester (a) 70.66 ml (4.83 mmol) of triethylamine was added to dry acetate. In 75 ml of nitrile, 0.82 g of the compound of Example 14(b) (2.20 mmol) benzotriazol-1-yloxytris (dimethylaminophosphoniu), benzotriazol-1-yloxytris Muhexafluorophosphate) 1.07 g (2.41 mmol) and Bali Solution of Luvaline methyl ester hydrochloride 0.64 g (2,4] mm4 mol) added to. , iL stirred for 4 hours, and 0.66 ml of triethylamine ( 4.83 mmol) was added and the reaction mixture was stirred for one wave. The solution was diluted with ethyl acetate. and washed sequentially with 2N hydrochloric acid, 5% sodium bicarbonate and water: sulfur After drying with magnesium acid and filtering, the solution was concentrated under vacuum. Eluate Flash chromatography using chloroform;methanol (40:1) as 466 mg of the title tripeptide (yield: 36 mg) was obtained as a white flaky solid. %) was obtained.

(3R,4S)-4-amino-2,2-difluoro-3-hydroxy-1-ox So-5-phenylpentyl-valylvaline methyl ester, hydrochloride b) Example 36 in 4Q ml of glacial acetic acid containing 450 mg of 10% Pd/carbon A solution of 466 mg (0.79 mmol) of the tripeptide (a) was saturated with hydrogen gas. After stirring overnight under a hydrogen atmosphere and filtering through a bed of Celite, the solution was was diluted with methanol and reacted with 100μ+ (1.0 mmol) of concentrated hydrochloric acid; The afi solution was concentrated under vacuum to give the crude tripeptide hydrochloride as a white crystalline solid. 390mg (i.

0%) was obtained.

(3R,4S)-4-(tert-buty)'v 7roxycarbonyl-(Q-buty)'v alanyl) amino-2,2-difluoro-3-hydrobin C1-oxo-5-phenylpentyl-valylvaline methyl ester C) Boc-Set (Bzl)-A in 6 ml of dry fATHF at -40″C Ia-Ala 380 mg (0.87 mmol and N-methylmorpholine 112 ml (0.87 mmol) of chill was added dropwise. After stirring for 30 minutes, N -Add 112 ml (0.87 mmol) of methylmorpholine, then add the solid The compound f of Example 36 (b) was added. The reaction mixture was gradually added to room temperature. The mixture was stirred overnight while warming. The solution was diluted with methylene chloride, 10% hydrochloric acid, 59 Continuously clean the leaking area with sodium hydrogen 6-hydrogen oxide and water and treat the leaking area with magnonium sulfate. dried, filtered and concentrated under vacuum. Chloroform meth/ as eluent - (30:1) was subjected to flash chromatography to produce a light yellow solid. 636 mg (83%) of the title hexapeptide was obtained.

4, 411 (2H, s), 4.22 (4H, ml, 3.62 (3 ni, sl, 3. '57 (13, m>, 2.98 (11 (, m), 2.71, (LH, m), 2.17 (2!(, ml, 1.4i( 9H, s), 1.1! l　(3HB d; J-6.5 Hz) / 1.03 (3H, d; J-7.1 iz ), 0.92 +12H/m) -MS (E’AB +): to/ Z-877 (M+H”) (3R,4S)-4 (tert-butyloxy cycarbonyl-serylalanylalanyl)ami/-2,2-difluoro-3-hypolymer Droxy-1-oxo-5-phenylpentylvalylno<phosphorus methylester d ) Example 36 (C) Li hexapeptide 13 mg (15 mm I7 mol) Acetic acid: methanol C5: 1) Dissolve in 6 ml and add 10% Pd/carbon to the solution. (13 mg) to saturate the mixture with hydrogen gas. Overnight under hydrogen atmosphere On a stirred stage, the solution was filtered through a bed of celite and condensed under vacuum to form a white crystal. 11.5 mg (99%) of the compound was obtained as a crystalline solid.

NMR (CDC13/CD30D): δ7.2415Hz m)t 4.1B (6H/mL 3.77 (1=, B).

3.65 +3)! , s), 3.52 (IH, ml, 3.09 (L H, ml, 2.87 (LH, zl, 2.03 (3H, m), 1.3 9 (9H, s), 1.31 (3H, d, J-5, 94Hz), 1.06 +33. and B J-6, 6HzL 0-90 (12H1m) MS: m/z 787 Example 37 (3R,43)-4-(serylalanylalanyl)amino-2,2-difluoro -3-hydroxy-1-oxo-5-phenylpentyl-valylvaline methyl ester Preparation of ster, trifluoroacetate 2 drops of TFA of Example 36(d) (Added to 1.1 mg (1.4 mmol) of protective hexapeptide. After 2 hours, A sieve was passed through the mixture and the excess TFA was evaporated. diethyl ether 3 a was added and evaporated three times in the same manner. The residue was dried under vacuum.

Example 38 4 (tert-butylox7carponylserylalanylalanyl)amino -2,2-difluoro-1,3-diogyuso 5-phenylpentyl-valylbari Preparation of 4-(tert-butyloxycarbonyl-(0-ben) methyl ester 7-2,2-difluoro-1,3-dioxyl) serylalanylalanyl)ami So-,5-phenylpentyl-valylvaline methyl ester a) Dry IDMF 2m 102 mg of α,α-difluoroalcohol of Example 37 (0.12 mmol) Mol) and Dess-Martin Belaiosina 7 (Dess-Martin TFA to a solution of 247 mg (0°58 mmol) of 43 μl (0.58 mmol) was added dropwise. After 16 hours, the solution was diluted with ethyl Diluted with ether, 320 ml of saturated aqueous NaHCl and 10% aqueous thiosulfate Pour into the mixture 7 ml of sodium. The mixture was stirred vigorously for 1 hour and then diluted with acetic acid ethyl chloride. Extracted with chill. The combined extracts were dried over magnesium sulfate, filtered, and dried under vacuum. It was concentrated. Using a gradient eluent of 0-30% methanol/chloroform The title α,α-difluor was subjected to flash chromatography as a white solid. 70 mg (yield 69%) of oc-ketone was obtained.

NMR (CDC13/CD30D): δ7.27 (log, ml, 4.5 0 (2H, 51, 4, 34 (5H.

ml, 3.71 +31. s), 3.66 (2H, rrl) , 3.25 (1M, ml, 2.99 (1VmL 2.24 (2H,, m), 142 (91,s), 1.34 (3H, ml, 1.08 (3H, rn), 0.X2 (12L m).

MS (rAB 10): m/z 875 (M+H”).

4-(tert-butyloxycarbonyl-serylalanylalanyl)amino -2,2-difluoro-1,3-dioxo-5-phenylpentyl-valylvari methyl ester b) Protected compound of Example 38(a) in 4 ml of glacial acetic acid containing 10% Pd/carbon. A solution of 15 mg (17 micromol) of the subpeptide was saturated with hydrogen, and a hydrogen atmosphere was created. The mixture was stirred under atmospheric pressure overnight. The mixture was filtered through a bed of Celite and concentrated under vacuum. did. Fluid using gradient 1 eluent of 0 to 50% methanol/licc form Rush chromatography produced the title debenzylated product as a white crystalline solid. 9 mg (67%) of the product was obtained.

Example 39 Ac-Ser-Gln-Asn-Tyr-Pro-Val-Val-NHt Production: 120 mg of the title compound was obtained in the same manner as above.

TLC: (n-BuOH/HOAc/H*0 4:1:1) Rf=O134 :(n-BuOH/EtOAc/HOAc/H,O11:1:1) Rf=0 ．． 66; HPLCk'=3. 0 (Hamilton PRP-1, CHsCN/ H, Olo, 1% TFA, 10%-40% CHsCN-10 min, linear gradient 1, 5 m01 min); FAB-MS: m/z 847.6 (M-i -H'') In the above method, converting Boc-Ser (Bzl) to Boc- Ac-Thr-Gln-Asn-Ty by changing to Thr(Bzl) r-Pro-Vat-Vat-NH7 was obtained.

Example 40 Ae-Set-Ala-Ala-Tyr　-Pro-Vat-Val-NH, Production: 95 mg of the title compound was obtained in the same manner as above.

TLC: (n-BuOH/HOAc/HtO4:1:1) Rf=0.7 2; HPLCk'=2.8 (Hamill) 7PRP-1, CH, CN/Hto1 0.1% TFA, 5% to 40% CHsCN, 15 minutes, linear gradient, 1, 51+1 (1/min); FAB-MS 2 m/z 747 (M+H’) Actual Example 41 Ac-Set-Gin-Asn-Tyr-D-Pro-Val-Val-NH, Manufacturing of 260 mg of the title compound was obtained in the same manner as above.

TLC: (n-BuOH-HOAc-HtO4:1:l) Rf=0 ．． 15:(n-BuOH-HOAe-H, 01:1:1) Rf=0.54; (n-BuOH-EtOAc-HOAc-H, 01:l:11) Rf=0.7F AB-MS: m/z 847 (M+H”) Amino acid analysis (0,0O5% Hydration at 160'C in HCl2/TFA 2:1 with w/v phenol for 11 hours. Decomposition): Asp, 1.00+Ser+Q.

75: Glu, 0.99: Pro, 0.99; Val, 0.96; Tyr, 1゜1゜ Example 42 Dns-3er-Gln-Asn-Tyr　Pro-Vai-Val-NHt Preparation of 56 mg of the title compound was obtained in the same manner as above.

TLC: (n-BuOH--HOAc-HtO4:1:1) Rf=0. 1HPLC (Hamilton PRP-1, CH, CN/HtO10, 1% TFA, 5 ~40% CH,CN gradient, 15 minutes), k'3.32FAB-MS: m/z 11038 (+H)” Amino acid analysis (6N HCl2, 11 0'C 124 hour hydrolysis): Asp, 1.00+ Ser, 0.17: Glu, 1.00; Pro, 1.00: Val, 1.38: Tyr, 1.78 Example 43 Production of Ac-3er-Gin-Asn-Tyr-Pro-Val-Arg-NHz Preparation: 61.5 mg of the title compound was obtained in the same manner as above.

T L C: (n-B uo H-pyridine-HoAc-H, 04:1: 1) Rf=0.43; (n-BuOH-EtOAc-HOAC-HtO1: 1: 1: 1) Rf=0.34; HPLC (Hamilton PRP- 1. CH3CN/H, Olo, 1% TFA, 5-40% CHs　CN glazier 15 minutes), k’! , 94FAB-MS: m/z 904 (M+H) "Amino acid analysis (110'C 118 hour hydrolysis in 6N HCQ): As p, 1.00; Ser, 0.98: Glu, 1.00; Pro, 0 ．． 88; Val, 0.56: Tyr, 0.86; Arg, 0.67 real Example 44 Preparation of Ac-Set-Gln-Ala-Tyr-Pro-Val-Val-NHt Preparation: 85 mg of the title compound was obtained in the same manner as above.

Gradient HPLC: 5-40%, 0.1% TFA (CH, CN), straight line , 15 minutes, k'=2.61 TLC: (B:, A:W 1: 1: 1), Rr=0.6 7 +FAB-MS: IIl/z 804 (MH)” Amino acid analysis (H Cl2: TFA 2: 1.0.005% w/v phenol, 160 "C11 hours): Ser, 0.89; Glu, 1.00: Pro, 1 ．． 04; Ala, 1.00; Val, 1.75; Tyr, 0.89 ; Peptide content = 98.94% Example 45 Preparation of Ac-Ser-5er-Asn-Tyr-Pro-Vat-Val-NHt Preparation: 78 mg of the title compound was obtained in the same manner as above.

Gradient HPLC: 5-40%, 0.1% TFA (CH3CN), straight line, 15 minutes, k' = 2.58 TLC:Rf=0.70. B:A:W 1: 1: 1;FAB-MS : m/z 806 (M+H)' Amino acid analysis CHCQ:TFA 2: 1, O, OO 5% w/v phenol, 160°C 11 hours): Asp, 1.00; Ser, 1.59: Pro, 0.90; Val, 1 ．． 91; Tyr, 0.91: Peptide content = 80.6% Example 46 Ac-3et-Arg-Asn-Tyr　Pro-Val　Val-NHt Preparation of 87 mg of the title compound was obtained in the same manner as above.

Straight line, 15 minutes, k' = 2.64 TLC: (B:A:W 1: l: 1), Rf=0.60; FAB- MS: m/z 875 (M+H)” Amino acid analysis: HC (/T F A /Phenol, Asp+1.00; Ser.

0.80: Pro, 0.83: Val, 1.85; Tyr, 0.94; Arg.

0.97 Peptide content = lO1.3% Example 47 Ac-3er-Lys--Asn-Tyr-Pro-Val-Val-NHt Production: 87 mg of the title compound was obtained in the same manner as above.

Gradient F (PLO: 5-40%, 0.1% TFA (CH, CN), straight line , 15 minutes, k' = 2.47 TLC: (B:A:W'1:1:1), Rr=0.60;FAB-M S: m/z 847 (M-I-H)” Amino acid analysis: (HCl2: TFA, 0.005% W/V 7 27-L.

160″C11 hours) Asp, 1.00; Ser, 0.81: P ro, 0.95; Val, 1.87; Tyr, 1.00; Lys, 0.84 peptide content = 92.3% Example 48 Preparation of Ac-5er-Glu-Asn-Tyr-Pro-Val-Val-NHt Preparation: 77 mg of the title compound was obtained in the same manner as above.

Gradient HPLC: 5-40%, 0.1% TF-A (CH, CN), direct Line, 15 minutes, k' = 2.72 FAB-MS: m/z 848 (\4÷H).

Amino acid analysis: HC (/TFA/fer/-fer, Asp, 1.06; Se r.

0.84; Glu, 1.00; Pro, 0.88: Ala , 1.00; Vat.

1.85; Tyr, 1.00: Contains peptide! =94.3% Example 49 Preparation of Ac-Ser-Ala-Asn-Tyr-Pro-Vat-Val-NHt Preparation: 86 mg of the title compound was obtained in the same manner as above.

TLC: (B:A:W 1:1:1), Rf=0.70; Gradient HPLC: 5-40%, O, 1% TFA (CH, CN), straight line, 15 minutes, k’ = 2.62 FAB-MS: vn/z 790 (MH)” Amino acid analysis: HCf2/ Tl”, A/phenol, Asp, 1.00; Ser.

0.81+ Pro, 0.94; Ala, 1. OO; Val, 1.8 7: Tyr.

0.76: Peptide content = 72.77% Example 50 Ac-3et-Gln-Ser-Tyr-Pro-Val-Val-NH, Production: 87 mg of the title compound was obtained in the same manner as above.

TLC: (B:A:W 1: l: 1), Rr=0.68; Graje HPLC: 5-40%, 0.1% TFA (CHsCN), linear, 15 minutes , k'=2.51 FAB-MS: m/z 820 (MH)” Amino acid analysis: HCff/T FA/Funi/-L, Ser, 1.60; Glu.

1.00; Pro, Q, 91; Val, 1.83: Tyr, 0 ．． 94; Peptide content = 84% Example 51 Production of Ac-3et-Gin-Arg-Tyr-Pro-Val-Val-NH Preparation: 89 mg of the title compound was obtained in the same manner as above.

TLC: (B:A:W 1:1:1), Rf=0.66; Gradient H PLC: 5-40%, 0.1% TFA (CHsCN), linear, 15 minutes, k' =2.58 FAB-MS: m/z 889 (M=H)” Amino acid analysis: HCI2/ TF A/Funi/-L, Ser, 0.80; Glu.

1.00: Pro, 1. OO; Val, 1.88; Arg, 1.0 1; Tyr.

0.81; Peptide content = 85.33% Example 52 , Ac-5er-Gin-Lys-Tyr-Pro-Val-Va I-NH, Manufacturing of Protected peptidyl resin Ac-3et (Bzl)-〇1n-Lys　(Cl　 z)-TYr (BrZ)-Pro-Va 1-Va I-BHA'1.5mi Rimol was prepared as described above, and 83 mg of the title compound was obtained by the method described above. Ta.

TLC・(B:A:W 1+1:1), Rf=0.69 gradation xn) HPL C: 5% to 40%, 0.1% TFA (CH, CN), linear, 15 minutes, k'= 2.48 FAB-MS: m/z861 (M=H)” Amino acid analysis: HCQ/TF A/Fe/-Fer, Ser O, 79, Glul, OO, Pro O, 94, Vat 1.85, Tyro, 96, Lyso, 90 Peptide content is 7 1.26% Example 53 Ac-Se r -G l n-Asp-Ty r-P ro-Va I-'a Production of l-NH, HJ peptidyl resin Ac-5e r (Bz I)-Gln-Asp (OBz I)-Tyr (BrZ)-Pro-Va l-Va I-BHA 1.5 mmol was prepared as described above and the title compound 8. 4 mg was obtained.

TLC: (B:A:W 1:1:1), Rf=0.73 gradient HP LC: 5% to 40%, 0.1% TFA (CH, CN), linear, 15 minutes, k' =2.63 FAB-MS+m/z 848 (M÷H).

Amino acid analysis: HCQ/TF-A/7. Knoll, Asp 1,02.5er 0.79, Glul, OO, Proo, 97, Val 1.36, Tyro, 82 Peptide content is 79.13% Example 54 Ac-Asp-Glr+-Asn-Ty r-Pro-Va 1-Va l-N Production of Ht Protected peptidyl resin Ac-, Asp (OBz I)-Gln-, Asn-Ty r (BrZ)-Pro-Va 1-Va 1.5 mmol of 1-BHA was added before 234 mg of the title compound (81,2 %) was obtained.

TLC: (B:E:A:W l:1:1:1) Rf=0.77; (B :P:A:W+ 13:10:3:12) Rf=0.56FAB MS 2m/z 875 (M ÷H).

HPLC: 4.5mmx25cm Altex Ultrasphere 5 m ODS.

UV detection at 220 nm, 5-40% acetonitrile water-aeration for 15 minutes. 7 nitrile 〇, 1% TFA gradient, k’=3゜Incratic HP LC: 85:15 water-acetonitrile 10, 1% TFA, ni'=4.77 Amino acid analysis: (HCQ/TFA containing 0,005% W/V phenol Hydrolyzed in 2:l at 160'C for 1 hour) -AST) 2.05, Glu 　1.00. P r o 1.03, ＼’a l 1.94, Tyro, 84 Example 55 Ac　-Hi　-G　n-As　n-Ty　r-P　ro-Va　I- Production of Va I-NH, Protected peptidyl resin Ac-His(Tos)-GIn-Asn -Tyr (BrZ) -Pro-Va I-Va 1-BHA 1.5 mm 48 mg (55 mg. , 2%).

TLC: (B:P:A:W 15:10:3:12) Rf=0.51: ( B:A:W 1:1:1) Rf=0.71HPLC: 4.5mmX25cm Altex Ultrasphere 5m ODS.

UV detection at 220 nm, 5-40% acetonitrile water-aeration for 15 minutes. Setonitrile-〇, 1% TFA gradient, k'=4.94 Incratic HPLC: 85:15 water-acetonitrile-〇, 1% TF A, ni' = 3.76 FAB MS: m/z 897 (M+H)'7 Mi/Wt analysis: (160 Hydrolyzed for 1 hour at °C) Aspl, 03, Glul, OO, Prol, 13, V a　I　2.00. Ty r 1.97, Hi S 1,28 Example 56 Ser-Gin-Asn-Tyr-Pro-Va I-Va l-NH, Manufacturing of Protected peptidyl resin Boc-3er (Bz1)-Gin-Asn-Tyr 1.5 mmol of (BrZ)-Pro-Va I-Va 1-BHA was added to the 188 mg (83.5%) of the title compound was prepared using the method described above. Obtained.

TLC: (B:E:A:W 1:1:1:1) Rf=0.68; (B:P:A :W:　15:10:3:12) Rf=0.54HPLC: 4.5mmx2 5Cm Altex Ultrasphere 5m ODS, 220nm UV detection in water-acetonitrile of 5-40% acetonitrile over 15 minutes Ryl-0,1% TFA gradient, k'=4.89 Incratic HPLC: 85: 15 Water-acetonitrile-〇, 1% TFA, to’-2.78 FAB MS: m/z 80.5.6 (M’-H)” Amino acid analysis: (16 in HC1!/TFA2:l containing 0,005% W/V phenol Hydrolyzed at 0°C for 1 hour) Aspl, 03.5er0.78, Glul, 0 O5Pro0.88, Va I 1.93, Tyro, 66 In the above sequence, 3-benzyloxy instead of Boc-Se r (Bz l) The desamino peptide 3-hydroxypropanoic acid was replaced with ci-propanoic acid. Obtain onylglutaminyl asparaginyl tyrosyl brolyl valyl valine amide .

Boc-D-3e instead of Boc-Se r (Bz I) in the above sequence 　r　(Bz　l), D-Ser-Gln-Asn-Tyr-Pr o-Va ] -Va l -NH, is obtained.

Example 57 Ac-Ser-Gin-Gly-Tyr-Pro-Va 1-Va 1-NH, Manufacturing of Protected peptidyl resin Ac-5er (Bzl)-Gln-Gly-Tyr (B rZ) -Pro-Va 1-Va 1.5 mmol of 1-BHA as above 145 mg (72.5%) of the title compound was obtained using the method described above. Ta.

TLC: (B':E:A:W l:1:l:1)Rf=O163; (B:A: V/ 4:1:1) Rf=0.40HPLC: 4.5mmX25cm A ltex Ultrasphere 5m ODS.

UV detection at 220 nm, 5-40% acetonitrile in water for 15 minutes -Acetonitrile-○, 1% TF A gradient, k゛=4.56 Incratic HPLC: 85:15 water-acetonitrile-0.1% TF ASk' = 3.46 FAB MS: m/z 790 (St+H)” Amino acid analysis: (0 ,003%W/V in HCC/TFA2:1 containing phenol, 160″ Hydrolyzed at C for 1 hour) 5er0.71, GIuo, 97, GIY 1. 00. P　　　　1.02, Va　　1.70゜Tyro, 76 Example 58 Ala-5er-Gin-Asn-Tyr-Pro-Va　-Val-NH Manufacture of t Protected peptidyl resin Boc-Ala-Ser (Bzl)-Gin-Asn-T yr (BrZ)-Pro-Va 1-Va 1-NHA was prepared as described above. Then, 185 mg (75.5%) of the title compound was obtained by the method described above.

TLC: (B:E:A:W 1: 1: 1: 1) Rf=0.6 4; (B:P:ASW 15:10:3:12) Rf=0.60HPLC: 4.5mmX25cm Altex Ultrasphere 5m O D.S.

UV detection at 220 nm, 5-40% acetonitrile in water for 15 minutes -Acetonitrile-0.1% TFA gradient, k'=3.86 Incratic HPLC: 85:15 water-acetonitrile-0.1% TF A, ni' = 2.71 FAB MS: m/z 876 (\4÷H) Amino acid analysis: (0 ,005%W/VHCl2/TFA2 containing phenol: 1 in 1 Hydrolyzed at 60'C for 1 hour) Aspl, 0O1Se rQ, 69, GIuo, 9 8, ProO, 95, Alao, 99, Va I 1.91, Tyro, 94 Example 59 Ac-3er-Gln-Asn-(4'NO,)Phe-Pro-V a l -Val　-NHt production Protected peptidyl resin Ac-5e　(Bz　I )-Gln-Asu-(4゛　NOx)Phe-Pro-Va　-Va　1 - 1.5 mmol of BHA was prepared as described above and marked by the procedure as described above. Compound 701.8 rng was obtained. Yield 80% ]% HOAc as eluent, 100 mg of the crude peptide was purified by Sephadex G. Purification by gel filtration on -15 yielded 68 mg. Preparative HPLC (PRP -1semi prep, 18% 0.1% TFA (CH, CN), 6m A further 33 mg portion was purified by C/min) to obtain 22 mg.

TLC: (B:A:~’ 1:1:1), Rf=0.34 Clazin)H PLC: 5% to 40%, 0.1% TFA (CH, CN), linear, 15 minutes, k'=3.18 F A B-MS: m/Z 876 (M÷H).

Amino acid analysis: (HC containing 0,005% W/V phenol (/TFA 2+1, 160’C” 1 hour hydrolysis) Aspl, 02.5er0.81 , Glul, 00. Proo, 90. Vat2.00, NidoaPheo, 91 Peptide content 103.67% Example 60 Dns-Arg-Ala-Ser-Gln-Asn-Tyr-Pro-Va 1 Production of -Va l -OH Protected peptidyl resin Boc-Arg (Tos)-A la-3er(Bz　])-]Gin-Asn-Tyr(BrZ)-Pro-V a1-Va I-Merryfield resin was produced as described above, and as described above. 321 mg of crude peptide was obtained by this method.

Using 1% HOAc as eluent 1, 2°6X7O of Sephadex G-15 A 100 mg portion of the crude peptide was purified by gel filtration on a cm column. suitable The relevant fraction was lyophilized to obtain 74.6 mg.

TLC: (B:A:W 1: l: I) Rf=0.84HPLC: ( Hamilton PRP, -1, CH3CN / H1010, 1% TFA, 5-40% CH, CN gradient, 15 minutes), k' = 3.6 1 FAB-MS: m/z 1266CM+H)'Amino acid analysis: (0,○05% 1 hour at 160'C in HCl2/TFA2 containing W/V phenol. Water splitting): Asp, 1.00; Ser, 0.31; Glu, Q, 91; Pro , 0.99:Ala, 0.96;Vat, 1.96;Tyr, 1.19:Arg , 0.85 Example 61 Ac-Se r-Gin-Asr+-Tyr-Pro-Va I-Va l-A Production of sn-NHZ Protected peptidyl resin Ac-5er (Bzl)-Gin-Asn-Tyr (B rZ)-Pro-Va l-Va 1-Gl n-A, sn-NHA 1 Millimoles of the title peptide 712 were prepared as described above, and the title peptide 712 was prepared as described above. ．． 9 mg was obtained.

TLC: (B:A:W 11:1) Rf=0.61 HPLC: ("Milton ( Hamilton) PRP-1, CH, CN/H, olo, 1% TFA, 5-4 0% CH, CN gradient, 15 min), 2,17 at k' FAB-MS: m/z 1089 (M1H)” Amino acid analysis: (O100 1 hour at 160'C in HCC/TFA 2:1 containing 5% W/V phenol. hydrolysis): Asp, 1.81; Ser, 0.97; Glu, 2.00; Pr o, 0.88; Vat.

1.23; Tyr, 1.81 Example 62 Ac-Ala-Thr-Leu-Asn-Phe-Pro-11e-Se r- Production of Pro-[1e-Glu-NH, protected peptidyl resin Ac-, AIa-T hr (Bz I)-Leu-Asn-Phe-Pro-I 1e-3e t (Bz　])-Pro-1　]e-Gl　u　(OB　z　1)-NWA 1 mmol of peptide 2 was prepared as described above, and the title peptide 2 was prepared by the method described above. :1', 6 mg was obtained. HPLC: (Hamilton PRP- 1, CHsCN/H, C10, 1% TFA, 5-40% CH3CN gradient 15 minutes), k'=2.94 FAB-MS: m/z 1241 (M-;H)' Example 63 Ac-3er-Gin-Asn-Tyr-Pro-Val-NH! Manufacture of In the same manner as above, 101.3 mg of the title compound was obtained.

TLC: (B:A:W 4:1:1) Rf=0.65HPLC: (Hamilton PRP-1, CH, CN/H, Olo, 1% TFA, 5-40 % CH, CN gradient end, 15 minutes), k.

~2. 08 FAB-:vlS: m/z 748 (MH)” Amino acid analysis: (, HCQ/TFA containing phenol of 005% ☆/- 160 in 2:1 'c, 1 hour hydrolysis): Asp, 1. 00; Ser, 0.55; Glu , 0.98+Pro, 1.19; Val, 0.9: v: Tyr, 0. 99 Example 64 Ac-Ser-Gin-Asn-Tyr-Pro-Val-Val-Arg-N Manufacture of H. 79 mg of the title compound was obtained in the same manner as above.

TLC: (B:E:A:W l・1:1:1) Rf=0.46HPLC: (, Altex Ultrasphere-C18, CH, CN/H, Olo, 1% TFA, 5-40% CH, CN gradient end, 15 minutes), to’ =3.71 FAB-MS: m/z 1003 (M'-, H)' Amino acid analysis: (in 2:1 HCQ/TFA containing 1,003% w/v phenol, 160'C, 1 hour hydrolysis): Asp, 1. 00; Ser, 0.73; Glu, 0.98; Pro, 0.98; Val, 1.84; Tyr, 1. 0 8: Arg, 0. 99 Example 65 AC-Ala-Gin-Asn-Tyr-Pro-Val-Val-NH, Preparation: 235 mg of the title compound was obtained in the same manner as above.

TLC: (B:E:ASW 1:l:I:1) Rf=0.69HPLC: (Altex Ultrasphere-C18, CH*CN/) (tolo, 1% TFA, 5-40% CH, CN (D'j'; end, 15 minutes), to ’=4.48 FAB-MS: m/z 831 (M+H)”7mi/acid analysis: (, 160 in HCQ/TFA 2:1 containing 0.005% W/V phenol "C11 hour hydrolysis): Asp, i, 00: Ala, 0.99; Gl u, 0.98: Pro, 0.95; Vat, 1.97; Tyr, 1.85 Example 66 Ala-Ala-Glu-Asn-Tyr　-Pro-Val-Val-NHz Preparation of 50 mg of the title compound was obtained in the same manner as above.

TLC: (B:E:A 1:1:1) Rf=0. 54HPLC: (Al Ta Nox Ultrasphere-C18, CH3CN/H!　O／　0　　、 1% TFA, O~40% C83CX no gradient, 15 minutes ), ni’=4.67 FAB-MS: m/z 947 (M+H)” Amino/acid analysis: (, 005% ★/V HCQ/TFA containing phenol 2:1, 160 Hydrolysis for 11 hours at °C): Asp, 1. 00; Ser, 0.61; Ala , 1.52; Glu, 0.99; Pro, 0.86; Val, 1. 90:Ty r, 0. 98 Example 67 Ac-Cys-Gin-, Asn-Tyr　-Pro-Val-Vat-N　H , 188 mg of the title compound was obtained in the same manner as above.

TLC: (B:E:A:W 1:1:1:1) Rf=O167HPLC: (Altex Ultrasphere-C18, CH, CN/Hto10.1% TFA, 5-40% CH, CN gradient, 15 minutes), to' = 3. 7 FAB-MS: m/z 863 (M=H)” Amino acid analysis: (0 Oxidized with performic acid for 54 hours, then containing 0.005% w/v phenol. Do H Cf! / T F A 2: Hydrogenation in 160°C for 11 hours Solution): Cys S OsH, 0,98; Asp, 1.03: Glu, 1.　 00;Pro, 1. 13; Val, 1. 94; Tyr, 1. 78 fruits Example 68 Ac-Set-Gln-Asn-Tyr-Δ3-pro-Val-~’al-N Manufacture of H. 41 mg of the title compound was obtained in the same manner as above.

HPLC: (Altex Ultrasphere-C18, CH, CN/H, 01 0.1% TFA, 10-40% CH, CN gradient, 30 minutes), ni’=3.　○○ F A B MS: m/z 803 (M”, H)’Amino acid content Analysis・(HCQ/TFA containing 0.005% w/v phenol 2:1 Medium, 160'C, 1 hour hydrolysis>:Asp, 1. 00: Ser, 0.　 66: Glu, 0. 95: Val, 1. 88; Tyr, 0. 95 Examples 69 Ac-Ser-Gln-Asn-Tyr-Pro-Val-Val-NHt Production: 138 mg of the title compound was obtained in the same manner as above.

HPLC: k’=4.17 (Beckman Ultrasphere-○DS, 5%CHsCN10.1%TFA to 40%cH,cN10°1%TFA, 15 minutes) MS (F A B): m/z 776 (M: H) Amino acid content Analysis::Asp 1.00;Gly 1. 03:Pro 0. 90;V al 1. 88; Tyr 0. 88. Ser 0.68 Example 70 2-Asparaginylamine/-3-phenyl-propyl-prolyl valyl varinea Manufacture of Mido 81.1 mg of the title compound was obtained in the same manner as above.

HPLC: (Hamilton PRP-1, 5~40%CH, CN10゜1%T FA, 15 minutes) k’=2.47 FAB-MS: m/z, 560.5 (M2H)” Example 71 (4S, 5S) 5-ami/-4-hydroxy-1-oxo-6-(4-hydro Roxyphenyl>hexyl-valylvaline methyl ester, hydrochloride (4S, 5S) 5-(tert-butoxycarbonyl)amino-4-hydro xy-1-oxo-6-(4-benzyloxyphenylhexyl-valylvaline methyl ester a) Example 35 (a) compound (1.08 g, 1.70 mmol) in methanol Sodium borohydride (32 mg, 0°85 mmol) was added to the solution at O'C. ) was added − times. After 30 minutes, the reaction was complete as shown by TLC and 5% HCg in water The mixture was diluted with a solution and extracted three times with CH and CQ.

The combined organic layers were dried (Mg5O,), filtered, evaporated under reduced pressure, and the isomers were mixed. The product was obtained as a white solid (1.05g, 97%). The isomer was heated for 1″×25 minutes. m　Pass through HPLC silica column, 40:ICH, C4l.

:MeOH at a rate of 20+Ill/min. First elutes at 14.5 minutes The desired (4S) isomer was collected and evaporated to give the title compound as a white solid ( 295 mg, 34%).

9FR (CDC13): δ7.43 (5Ht rag, 7.02 (4H t caL 6.40 +LH,cri.

6.32 (l)! , d), 5.04 (2H,s), 4.50 (2H, m), 4.26 (21 (, rn), @3.7C (3H, 5) / 2.71 (23!, ml, 2.43 (2H , m>t 2. LL (2H, ml, 1.39 i9::+ s), 0.91 (12H, ml.

MS ffAri: rII/2 642.4 CM+H)”.

(4S, 5S) 5-amino-4-hydro-gyu-shi 1-o-gyu-so 6-(4-ben Zylox7phenyl)hexyl-valyl, (phosphorus methyl ester, hydrochloride b) The compound of Example 71(a) (295 rg, 0.46 mmol) was added to the neat solution. It was dissolved in refluoroacetic acid (1-) and stirred for 3 minutes. The solution was evaporated under reduced pressure, Dissolved in MeOH, treated with concentrated HCQ (3 drops) and evaporated under reduced pressure to give the title compound. Obtained as a white solid.

Example 72 (4S, 5S) 5-(alanylalanyl)amino-4-hydroxy-1-o xo-6-(4-hydroxyphenyl)hexyl-, tel, acetate (4S, 5S)-5-(carbobenzyloquinalanylalanyl)amino- 6-(4-benzylquin)phenyl-4-hydroxy-(l-oxo)hexy rub-valylvaline methyl nismol a) carbobenzyloxyalanylalanine ( 56+ng, O, 19 mmol) in THF under an argon atmosphere at -40 Cooled to 'C.

Add N-methylmorpholine (28m!2.0.231 mmol) to this solution. Well, then add inbutyl chloroformate (23m12, 0.19 mmol). added. The reaction mixture was stirred for 15 minutes and then added with N-methylmorpholine (28d, 0.251 mmol) and then added the compound of Example 71(b) (10 0 mg, 0.173 mmol) was added. The resulting mixture was incubated at -40°C for 3 The mixture was stirred for 0 minutes, warmed to room temperature, and stirred for 16 hours. The mixture was mixed with 1:1 ethyl acetate. :CHtC (, diluted, 5% HCJ aqueous solution, 5% N a HCO3 aqueous solution and Washed sequentially with saturated Na(J) aqueous solution.The solution was dried and filtered. , and atrophied under reduced pressure. The solid residue (7M) was purified by 1”×25CIII HPLC Through Kagel force gel, 95:3:0.5 CHCC:+:λ1eOH:H, The title compound was obtained as a white solid (96 mg, 77%).

NMR (CD30D/CDC13): δ7.15 (LOH, ral, 5 ．． εO(4H+cjl, 4.50 (2:.

ri, 4.81 (23', + 51.3.90 (2!=, qll 3.1 5 fLHl m) 73.51 (3 =, iL3.36 + l right mL 2 ．． 59 (2H, m) + 2. iL (2H, rr+L 1.88 ( 3 formula = l, 1.48 (24ml, 1.10 +8H, ml, 0 ．． 76 +12:,, cjl.

MS (FAB): rn/z 81B, 3 (M+3E)”.

(4S, 5S) 3-(alanylalanyl)amino-4-hydroxy-1-o Xo-6-(4-hydroxyphenyl)hequinruvalylvaline methyl ester , acetate b) Compound of Example 72(a) (90n+s;, 0.11 mmol) in acetic acid To the solution was added 10% palladium on activated carbon (90 mg). balloons with hydrogen gas for 60 minutes, then the stirred mixture was bubbled through the solution for 60 minutes via It was kept under hydrogen atmosphere for 4 hours. filtering the suspension through a pad of celite; Withering under reduced pressure gave the i-hypertrophic compound as a white solid (74 mg, 98%).

NMR (CD30D/CDC13): δ7.06 (LH, d), 6.78 +4H, dd), 4.29 +LH,・dot 4-16 (IHlm), 4 ．． 03 (IHld), 3-60 (3G!r sl, 2-62 +2)! 1 m) -1J5 (7H, by: ml, 1.57 +2H, ml, 1.16 +6H, ml, 0.80 +124 d).

MS (rAB): m/z 594.4 (M+H door.

Implementation 1'de 173 (4S,35)-:v-alanylamino-4-hydroxy-1-oxo6-( 4-Hydroxyphenyl)-hebosilvalylvaline methyl ester, acetate (4S, 5s) 5-(carbobenzyloxyalanyl)ami/-4-hydro Oxy-1-oxo-6-(4-benzyloxy/phenyl) to beef silver methyl ester a) Using the method of Example N 72 (b), carbobenzyloxyalanylara Carbobenzyloxyalanine (34 mg, 0.15 mmol) was substituted for nin. , N-methylmorpholine (44m(,0.40 mmol.

(added in equal amounts twice), isobutyl chloride; formate (20 mC, 0°15 mC) ) and the compound of Example 7)(b) (79 mg, 0.137 mmol). The i-gyuhi compound was synthesized using : Purified by Kalani chromatography using silica gel, eluted with CHC. Obtain the title compound as a white solid? = (60 mg, 82%).

NK (fcD30D/CDC13): δ 7.13 flOH, m), 6.81 (4F,, dd), 4.92 i2W.

s), 4.70 +2H, s), 4.21 +IL rn), 3 ．． 96 (2)! , d), 3.76 (LM, ij.

3.55 (3H, sl, 3.34 (IL m), 2.61 (2 H, mL 2.12 (2Hz': aIr 1.69 + 3H, m), 1. 48 (2H, rnl, 1.04 +4H, d), 0.71 (12M, d).

MS (!’λB): m/z 747.3 (M÷H).

(4S,5s)-3-alanylamino-4-hydroxy-1-oxv-6-(4 -Hydroxy7znyl)-hexylneuryl, <IJn methyl ester, acetic acid salt The compound bHii'i is the compound of Example 73(a) added to the compound of Example 72(a). (60 mg) was synthesized in the same manner as compound 72(b), and 1 Gyuhi compound The product was obtained as a white solid (52 mg, 100%).

NMR (CD30D/CDC13): δ8.19 (LH, d), 7.95 (LH, d), 7.70 (IH.

d) p 6-78 (44dd),, 4.15 (2H1mat 3- 95 4114-m), 3-13 (IH/q)/3.58 (3H, s ), 3.43 + IH, ml, 2.66 (2H, ml, 2.21 (2 )! , t), 1.90 (3a, m), 1.53 <2Ht (ri, 1 ．． 36 +3H, d), 1.17 +2H, br)', 0.93 +2)! .

ml, 0.78 (12M, d).

MS CI’AB): m/z 523.2 (M+H)”.

Example 74 (4S, 5S) 5 (tert-butyloxycarbonylalanyl- 4-Hydroxy-l-oxo-6-(4-hydroxyphenyl)be/rubaly Preparation of Luvaline Methynystyl The title compound was obtained in the same manner as above (7. 5B, 7 Q%).

NMR (CD30D/CDC13): δ6.72 (4Pl, dd), 4 ．． 23 (IH, d), 3.65-3.92 (3L rn) t 3.54 ( 3L s), 3.38 (2H, mar 2.60 (2H, ml, 1.6 2-2.20 (5Ht mat 1.49 (3H, ff1), !, 26 ( 91 (, s), 1.04 (5H, mLO, 77 (12M.

d)・ MS (FAB): m/z 623.3 (M+H)”.

Example 75 (4S, 5S) 5-((β-alanyl)alanyl)amino-4-hydroxy -1-oxo-6-(4-hydroxyphenyl)hexyl-valylvaline methyl Production of ester, hydrochloride (4S, +S) 5-((carbobenzylo+/-β -alanyl)alanyl)amine-4-hydroquine-1-oxo-6-(4-hydro (roxyphenyl) carbobenruvaline methyl ester a) carbobenruo Quin-beta-alanine (23 mg, 0.810.5 mmol) and hydroquinoben Zotriazole (26 mg, 0.19 mmol) glue,',=IF(0,8 m12) solution, add synchrohequinlecarphodiimide (22mg, O, 105ml) mol) was added and the mixture was stirred for 20 minutes in a dark room. X-methyl mol to this solution Holin (32rhC10, 29 mmol) and the compound of Example 73(b) (5 0 mg, 0.0956 mmol) was added and the solution was stirred at room temperature for 6 days. diluted with CHC43, 10% HCQ, 5% XaHCOs aqueous solution and H2O Washed sequentially with The organic 1 was dried (Mg5O,), filtered and a white solid (44+ ng) was evaporated under reduced pressure. The product was subjected to 1" x 25cm HPLC Pass through a silica column and elute with 6:96 λ4eOH:CHCρ, and the title compound The product was obtained as a white solid (12 mg, 20%) 5NyR' (CD30D/C DC13): S 7. ! il <LH, dl, 7.30 (IH , d), 7.18 (TH, b: d), 0.80 (12Thr d)-MS (F'AB): m/z 72 8.3 (M+F,)”.

(4S, 5S) 5-((β-alanyl)alanyl)amino-4-hydrocytol -1-one-6-(4-human xyphenyl)hexyl-valylvaline methyl ester hydrochloride b) MeOH of the compound of Example 7.5 (aXl 1 mg, 0.015 mmol) Add 10% palladium on activated carbon (5ig) to the solution. balloons with hydrogen gas Bubble through the solution for 1 hour via , placed under hydrogen (latm) 72. The suspension was filtered through Celite. Then, a few drops of cone and HCQ were added. The 1 volume liquid was evaporated under reduced pressure and marked The compound was obtained as a white solid (lomg, 95%).

NFR (CD30D/CDC13): δ7.60 +1 formula d>:6.59 ( 4L dco, 4.06 ns.

t), 3-89 +2! (/m), 3.66 (14mL 3.44 (3 H/S) 73.27 (lHz ml.

2.72 (2H, mL 2.41 (5H, ml, 1.98 ( 2H, tl, 1.79 (3H, m), 1.32 (2d, mar 1 ．． 01 (l) I, m), 0-94 +3H-d) t 01o7 (12H, d) Example 76 (4S, 5S) 5-amino-4-human; to xy-1-oxo-6-phenyl Production of xyl-valylvaline methylnistil% salt Ht salt (4S, 5S) 5 (tert-butyloxycarbonyl)amino-4-hydroxy-1-oxy So-6-phenyl pentasilva+Hluvaline methyl ester a) Implementation 1? 'll 34 (a)'s own product (4.39 g, 8.61 mmol ) in MeOH (150r, L) at 0°C. , 33 g, 8.7 mmol) were added. Obtained 7: Warm the mixture to room temperature, Stir for 30 minutes. The mixture was poured into 5% HCC (500+n) and CHtC ( The combined organic layers were dried (X1 g So4) and filtered. Filtration and evaporation under reduced pressure gave a mixture of stereoisomers. The residue was diluted with 2% MeO) (/ Dissolve l5 in CHC (!s (70m12), filter, 2” x 2’+l silica column Purified by IPLC using 2% MeOH/CHCQ, I put it out. The first eluting desired (4S) isomer was collected, evaporated under reduced pressure, and labeled. The compound was obtained (1.23g, 27%).

NMR (CDC131: δ7.27 (5H, ml, 6.94 (l)!, d), 6.76 (1:(, dl, 5.LO(1-H,d), 4-51(I Hldd), 4-36 (IH-br tL 3-74 (3″:tsL 3 ．． 10-3.50 (3H, ml, 2.89 (2L d), 2.40 ( 2)! , ml, 2.24-2.00 (2H, it.

1.95-1.65 (2H, m), 138 (9H, s), 0.91 ( 12H, ml.

MS (FAB): m/z 536.2 (M+)! B.

(4S, 5S) to 5-amino-4-hydroxy-1-ox/-6-phenyl Xyl-valylvaline methyl ester, tLaj8b) Example 76 (a) Putido (1.0 g, 1.87 mmol) was treated with trifluoro-C acetic acid (5 moromi). , and the resulting mixture was stirred for 10 minutes.

Add meta/-ru (23+n(:), then ccr+c, HCC(0,20m Q,) was added. The solvent was evaporated under reduced pressure, and EL and O were added to the residue and evaporated again. , the compound was obtained as a white powder, which was used without further purification. Ta.

Nx (CD30Dl: δ7.30 (5J(,m)t 4.26 (2 H/m) t 3-69 (3h, s) -3-60 (IH, m), 3.34 +LH, ml, 3.07 +13i, dd), 2.88 (LH , dd), 2.40 (2H, ml, 2.08 (22m+, 1.80 (2H, ml, 0.94 (12H, ml.

MS (TAB): m/z 436.2 (M+H)”.

Example 77 (4S,,5S) 5-(alanyl)amino-4-hydroxy-1-oxo- Production of 6-phenylhexyl-valylvaline methyl ester acetate (45,5S) 3-(carbobenzyloxyalanyl)amino-4-hydro Quin-1-omoso-6-phenylhexyneruhalylhalin methyl ester a) Carbobenzyloxyalanyl (4) 3 mg, 1.85 mmol) Anhydrous T)(F(5:) solution, -40'C, under argon atmosphere, X-methyl Morpholine (Q, 24., (2,2 mmol), then isobutylchloride Roformate (0.240ml, 1.83 mmol) was added. Stir for 20 minutes After that, add N-methylmorpholine (0.24ffl12.2.2 mmol) In addition, the compound of Example 76(b) (0.83 g, 1.0 millimoles ) Add OTHF (10 mC) solution 1; The resulting mixture was warmed to room temperature and overnight Stirred. The mixture was dissolved in EtOAc and diluted with 10% HC(, 5% Xa, HCOs It was washed successively with aqueous solution and H,O, and then dried (MgSO,). filtration and A white solid was obtained by distilling off the solvent, and this 1t was diluted with 4% to 10% MeOH. / Furanone dichromatography eluting with a gradient of CHCIs solvents MFJ (0.973.g, 86% redundancy) NMR (CD30D): δ7.34C5H,rn), 7.23 (54r n), 5.09 +2H,! 1.4.32 (IH, d), 4. L9 (IL d), 4.09 (2H, ml, 3.71 <3H, s), 3.60 (1:!

ml, 2.85 j2F, t mL 2.30 (2H/mL 2. Yu O + 2L m), ! , 70 (2H, mgeki B 1.23 (3H, d), 0.94 (12H, ml.

MS (1’AB): m/z 641.3 +M+H)”.

(4S,,5S) 5-(alanyl)ami/-4-hydronine-1-Ogyuso 6-phenyl to bovine sylvalylvaline, methylnystyl acetate b) 1° of the compound of Example 77 (a) (969 mg, 1.51 mmol) 10% palladium on activated carbon in MeOH:HOAC (lδv2) solution (100 mg) was added. Hydrogen gas was introduced into the mixture and then water was added for 18 hours. It was kept under normal atmosphere. Pour the reaction mixture onto a celite bar.

Filtration through a tube and evaporation under reduced pressure afforded the Rituki compound as a white solid (0.8 4g, 98%).

N (CD30D): δ 7.25 (5H, ml, 4.32 (IH, d), 4.21 (YH, ci), 4. P0 (LH, b: t), 3.77 (lH, leaf; 6tr; q+, 3.71 (34s), 3.60 (l b:t), 2.96 (IH, dd), 2. ．． 83 (IH, dd>, 2.35 +2H, C1, 2, 08C26, i).

'1.96 (3H, ri, 1.70 (2H,!'n), 1.44 (3 H, d), 0.95 (12M,, d).

MS (TAB): m/z 507.2 (M+H1” Example 78 (4S, 5S) 5-(alanylalanyl)amino-4-hydroxy-1-o Production of xo-〇-phenylhexyluba 1, ゛ruhaline methyl ester (4S, 5s> 3-(carbobenzyloxyalanylalanyl)amino-4 -Hydroxy-1-okine-6-7phenylhexylvalylvaline methyl ester le a) Carbobenzyloxyalanyl (337 mg, 1.51 mmol) and X-methylmorpholine (0,1981Tli!, 1.80 mmol) without O To water THF (10mf2) solution, -40''C, under argon atmosphere, imbutirc. Roloformate (0°196m (,1,51 mmol) was added over 10 minutes. . After further stirring for 20 minutes, X-methylmorpholine (0°198m(!, 1.80 mmol), Life, 77(b) (D compound (0.84 g, 1.48 mmol) Added a solution of THF (2" OmC) in dimethylformamide (10v2) was added and the cold bath was briefly removed to facilitate stirring. The reaction mixture was diluted with 40% It was cooled again to 0.degree. C., slowly warmed to room temperature and stirred. The reaction mixture was %X1eOH/CHC13 (000rrlI2), H=O(200- ), 10% HC (: (200 m and H . The organic extract was evaporated under reduced pressure, and the residue was converted to 80:20:2 CHC (!, :MeOH:Hto (dissolved in 22m.

Volume of liquid: = 3 times; Discard, 90: 10: I CHCl2s:M By synchromatography, eluting with eOH:H,o (22 mQ) The crude product was obtained as a yellow-white solid (950a+g). this substance Solubilize to 5% 'v1eOH/CHCQxC7vmQ), filter, and add 8@Q each 5% Me○H by HPLC using a 1"X25e11 silica column. /CHCQ, to give the title compound as a white solid (72 lrAg, 68%).

NMR (CD30D/CDC13): δ7.35 +5H, rr, ), 7. 22 (5H, m), 5.12 +2H.

d), 4.36 fIH, ci), 4.29 +IHr q)/4.16 (2)! , m), 4.05 +13E, ml.

3j3 +3H, sl, 3.57 (IH, ml, 2.92 (IH, C 1d), 2.81 (LH, ddl.

2.32+2H,I? Il, 2.09 (2H, m), 1.72 (2H , ml, 1.36 (3H, c!+, 1.27 (3H, dl, 0.94 +12H, 2 d's).

MS (FAB): rn/z 712.4 (M+H1”.

(j5.5S)'5-(alanylalanyl)amino-4-(droxy-1-ox So-6-phenylhexylvalylvaline methyl ester b) Implementation PI 78 ( Compound a) (713 mg, 1.01 mmol of λ4eOH (45 mj) To solution 1α, 10% palladium on activated carbon (IOOmg) was added. the reaction Hydrogen gas was introduced into the mixture and then kept under hydrogen atmosphere for 48 hours. the mixture The material was filtered and the solvent was evaporated to give the title compound as a white solid (599 mg, 100% n.

NMR (CD30D): δ7.24 +5H, ml, 4.32 +2L m l, 4.21 (l town d), 4.03 (LH, br t), 3.94 ( IH, q), 3.71 (3B, sl, 3.59 (IH, br Cl, 2J6 (2H, 2L '99 dd's), 2.31 (2 town t), 2.07 (2H, ml, 1.68 (2H, ml, 1.52 (31(, d), 1.32 (3H, d), 0.94 +12H, i dd's).

MS (rAB): m/z 57B, 3 fM+H)".

Example 79 (4S, 5S) 5-(alanylalanyl)amino-4-hydroxy-1-o Production of xo-6-cyclohexylhexyl-valylvaline methyl ester Compound of Example 78(b) (5.4 mg, 9.4 mmol) in λIeOH (5 P to t < 10 mg) was added to the solution of Cool Hydrogenaeor (Parr Hydrogenaeor), 2 Shake for days. 3) Filter the kelp, distill off the solvent, and remove the gyuhi compound (white). Obtained as a solid (3.4 mg, 62%).

N Nagi (CD30D): δ4.42-4.15 (3H, zI, 3.97 (2H, ml, 3.71 (1, sl.

3.52 (IHtm), 2.35<2H, t), 2.10 (2H, m), z, a s(LM,brd).

1.80-1.15 (21H, m), i, 65 (121, m).

MS (FAB): m/z 5B4.4 (M+H)”.

Example 80 (4S, 5S) 5-(serylalanylalanyl)amino-4-hydroxy- 1-oxo-6-phenylhexyl ester, vinegar Production of acid salts (4S, 5s), 5-(carbobenzyloxy-(0-phenylmethyl) lylalanylalanyl)amino-4-hydroxy-1-oquinone-6-phninyl Hexylvalylnozaline methyl ester a) Carbobenzyloxy-(○be Serine (3 1 1 mg, 0.

945 mmol) and N-methylmorpholine (130+n+?, 1.20 mmol) Nibutyl chloroform was added to a solution of HF (20 mf2) containing mol) at -40°C. Add 10 molecules of Mayl- (123 mR, 0.945 mmol) at j':t. After stirring for an additional 30 minutes, the compound (+20 mg.0.900 mmol) was added to 2o% DλIF/THF (No. 25) by solubilization.

The resulting mixture was slowly warmed to room temperature and stirred for 20 hours. The mixture was converted into CHC The organic extract was diluted with Cs (100 m) and washed with Ht (twice), and the organic extract was evaporated under reduced pressure. I let it emanate. Solid residue (870 mgH±, 10% MeOH/CH Cj. , preloaded by furanosinic chromatography and still somewhat impure. Got quality. The product was dissolved in 4%\1eOH/CHC & 20mC) and filtered. , 4% M e O by HPLC using a I” x 25 cm silica column. H / C H t C Q- was eluted and further purified in 6 times, and the title The compound was obtained as a white solid (41s), 3.38 (IL m), 2.7 3 (IL dd), 2.63 (l)! , del), 2.10 [2H , m).

(4S,5S)　　　5-ECO-7dynylmethyl)serylalanylalanylnia Mi/-1-oxo-6-phenyl to bovine silvalylvaline methyl ester b) Example 80 (a) of the compound (409, mg, 0.461 mmol) 2:l　MeOH/HO,Ac (13m(i) A mixture of 1 on activated carbon 0% palladium (80 mg) was added. Hydrogen gas is introduced into the reaction mixture and then The mixture was kept under hydrogen atmosphere for 16 hours and filtered through Celite to reduce the Evaporation under pressure gave the title compound as a white solid (353mg, 94%).

NMR (CDC13): δ7.31 (5H, ml, 7.24 (5H, ml , 4.58 (2H, s).

4.42-4.18 (48, ml, 4.05 (IH, m>, 3 ．． 94 (IH, rn), 3.85-3.70 (QH.

m), 3.70 +3H, s), 3.58 (LH, m), 2.9L (II(, dd), 2.81. (iH, cid).

2.30 (2M, ml, 2.12 +2H, rr+), 1.98 (3H, 5), 1.72 (2H, ml, 1.R7 (3H, d), 1.22 (3H, d), 0.94 (12H, m l.

MS ffj’G): m/z 755.3 (M+H)”, ゛(4S, 5S ) 5-(serylalanylalanyl)ami/-4-hydroquine-1-oxo 6-Fninylhexylvalylno(phosphorus methyl ester, acetate) C) Compound of Example 80(c) (333 mg, 0.46 mmol) in XIeOH (20 mC) To the solution was added 10% If radium on activated carbon (85 mg).

Hydrogen gas was introduced into the mixture and kept under hydrogen atmosphere for 2 days. Acetic acid ( +mQ,) was added, followed by further addition of 10% Pd/C (1.50 mg). Furthermore After stirring under hydrogen atmosphere for 4 days, the reaction mixture was filtered through Celite and the solvent was removed. Evaporation under reduced pressure gave the title compound (273 mg, 88%) O NMR (CD30D): δ7.23 +5H, m), 4.43-4.18 (4H, m)' 4.05 (1 set = (2H, m), 1-40 (3H, d ), 1-25 (3H1d), 0-94 L12Hr m) -MS ffAB ): m/z 665.3 (M+H)”.

Example 81 (4S, 5S) 5 C(D-seryl)alanylalanylcoamino-4-hyron Doroki/-1-oxo-6-phenylhexylvalylvaline methyl ester manufacturing (4S,5S)-5-((carbobenzyloxy-D-seryl)alanylalani ) Amino-4-hydrotei l-oxo-6-phenylhexylvalyl Phosphorus methyl ester a) Carbobenzyloxo-D-serial in DMF (8a) (8゜4mg, 33mmo I) and hydroxybenzotriazole) 95 mg, 35 mmol) overnight, and then dissolved zinc clohexylcarbodiimide (7.2 m g, 35 mmol) was added and the mixture was stirred for 20 minutes. Example 78(b) Compound (20 mg, 35 mmol) was added, followed by DMF (7 drops). The mixture was diluted with 10% MeOH/CHCl, 10% HCI, 5% aqueous Na Washed sequentially with HCOs and water. The solvent is removed under vacuum and the residue is flash-crushed. Purified by chromatography and eluted with 10% MeOH/CHCIs 4.2 mg of the title compound was obtained (yield 14%).

N4 (CDC13/CD30D): δ7.26 (SH,s), 7.17 (5M, ml, 5.11 (IH.

+3H, (1), 1.24 (3H, (11,0,87+121.d).

MS (FAB): m/z 799.5 +M+H-.

(4S,5S)-5-((D-seryl)alanylalanyl)amino-4-hydro xy-1-oxo-6-7dynylhex/ruvalylvaline methyl ester b) 3.9 mg (4.9 mmol) of the compound of Example 81(a) in MeO82 2 mg of 10% palladium on carbon was added to the ml solution. Introducing hydrogen gas into the reaction mixture Then, it was maintained in a hydrogen gas atmosphere for 26 VJfi. , filter the mixture through Celite and reduce Evaporation under pressure gave 3.4 mg of the title compound as a white solid (98% yield).

NMR (CD30D): δ7.22 +58. ml, 4.25 (41, ml, 4.05 + IH, ff1), 3.82 (21, ml, 3.69 (3 ratio s), 3.55 (IL m), 2.90 (IL dd), 2. 78+IF,.

MS (FAB): m/z 665.4 (M+H).

Example 82 (4S,5S)-5-amino-6-(4-penzyloxo)phenyl-4-hydro Production of Roxy-1-oxo-hexyl-valyl valine benzyl ester hydrochloride (5S)-6-(4-pendyloquine)phenyl-5-(t-butoxycarbonylate) )amino-(1,4-dioxo)hexylvalyl valine benzyl ester a) 43 mg of the compound of Example 13c) at -40'C under argon atmosphere (0,1 16 ml of X-methylmorpholine (0.14 m mol) Then 13 ml of inbutyl chloroformate (0, I O+r+ rr+o I) was added. After stirring for 15 minutes, add N-methylmorpholine 16 Add m1 (0.14 mmol), then barylvaline benzyl ester salt Acid [51 mg 80.15 mmol) was added. The resulting mixture was allowed to rise to room temperature. Then, stir overnight. The reaction mixture was diluted with 5% HCI and extracted three times with CHCl. Ta. The extracts were combined and subjected to furanosinichromatography, l:25Me.

Elution with H:CHCl gave 62 mg of the title compound as a white solid (yield 100%). ).

NMR (CDC13): δ7.34 (IOH, ml, 7.0742H, d ), 6.89 (2H, d).

6.52 (IH, d), 6.31 (l town dl, 5.2 (1H, rrl , 5.16 (2H, dd), 5.02 (2H, s), 4.59 TIH , dd), 4.49 (IH, dd), 4.30 (IH, dd), 3. 08(4S, 5S)-6-(4-benzyloxy)phenyl-5-(t-butyloxy) Toxycarbonyl)amino-4-hydroxy-1-oxo-hexyl-valyl Valine benzyl ester b) Example 82 (a) Compound 61. mg (0,1 Boron hydride in 5:IMeOH:CH,C1,6rr+l solution of 0 mmol 5 mg (0.13 mmol) of sodium chloride was added, and the resulting mixture was heated to 20 Stir for a minute. The reaction mixture was diluted with 5% aqueous MCI and extracted several times with CH,CI. did. The extracts were combined, dried over MgSO4, filtered and evaporated under reduced pressure to a white solid. The title compound of the body 61. mg (100% yield).

NKL” (CDC13/CD30D): 67.35 (10H, ml, 7 ．． 12 (2J (, di, 6.90 (2H.

d), 5.17 (2H, dd), 5.02 (2H, s), 4.47 (1M, m), 4.17 (19 Kin et al #’rf tL 3.6B (1! , m), 3.53 (IH, m), 2.96-2.55 (2H, ml.

2.36 (2H, m), 2.19 (IH, m), 2.10-1.81' (2H, ml, 1.70 (IH, ml.

(4S,5S)-5-ami/=6-(benzyloxy)phenyl-4-hydro xy-1-oxo-hexylvalyl valine benzyl ester hydrochloride C) CHtCI of Example 82 (b) compound 60 mg (97 mmo I) Add 3 ml of trifluoroacetic acid Q to 22 ml of solution, and stir the resulting solution for 2 hours. did. Methanol and then 2 drops of concentrated hydrochloric acid were added and evaporated under reduced pressure. Remove the residue Trituration with alcohol gave 56 mg of the title compound as a white solid (yield 100 %).

NIQ (CD30Dl: δ7.36 (10H, m), 7.20 (2H , d), 6.97 (2) 1. d).

5.15 +2H, m), 5.07 (2H, s), 4.34 (IH, m l, 4.21 (IH, ml, 3.84-3.53 (IH, ml, 3. 44-3.20 i1 formula mar 2.97 (IH, ml, 2.80 (IH , ml.

(4S,,5S)-5-amino-6-(4-hydroxy)phenyl-4-hydro xy-1-oxo-hexyl-valyl valine benzyl ester hydrochloride C) Dissolve 50 mg of the compound of Example 82(a) in acetic acid and add 10% palladium on carbon. Stirred with 50 mg. Hydrogen gas was passed through the solution for 2 hours and the reaction mixture was placed in a hydrogen atmosphere. The mixture was stirred under air for 20 hours. The suspension was passed through a pad of Celite for 11 minutes under reduced pressure. The title compound was obtained by filtration.

Example 83 (4R3,3S)-5-(serylalanylalanyl)amino-6-(4-hydro xy)phenyl-4-hydroxy-1-oxo-hexyl-valyl valine hydrochloride manufacturing of salt (4RS, 5S)-5-[(t-butoxycarbonyl-(O-phenylmethyl) Serylalanylalanyl)amino]-6-(4-benzyloxy)-4-hydro Roxy-1-oxohexyl-valyl valine benzyl ester a) t-Butoxycarbonate in 1 ml of THF at -40'C under argon atmosphere Le-(0-benzyl)serylalanylalanine 46° 8mg (107mmo) 16.5 ml (150 mmol) of N-methylmorpholine was added to the solution of I), and then Then, 13°8 mt (107 mmol) of inbutyl chloroformate was added.

After stirring for 15 minutes, add 16.5 ml (150 mmol) of X-methylmorpholine. In addition, 56 mg (97 mmol) of the compound of Tweet Example 82 (C) A 73 ml solution of HFo was added. The resulting mixture was warmed to room temperature and stirred overnight. did. The reaction mixture was diluted with 5% MCI and extracted several times with CHCIt. extract liquid Combine and evaporate under reduced pressure, and evaporate the resulting solid residue.

It was purified by chromatography and eluted with 1:9 SieOH:CHCIs. 79.5 mg (yield: 87%) of the title compound was obtained.

d's) t 1-17 +3) 1,2d's), 0.96 +124 +l+1 )-MS (YAB): mlz 1037.4 (M÷H)+.

(4R3,5S)-5-(t-butoxycarbonylserylalanylalanyl)a Mino-6-(4-hydroquine)phenyl-4-hydroboxy-1-oxo-hexy Rubalyl valine b) 79 mg (76 mm) of the compound of Example 83 (a) o l) Part 3: 10% palladium in 1 MeOH: HOAc 5 ml solution 45 mg of carbon was added. Hydrogen was introduced into this mixture and heated at latm Hz for 24 hours. held for a while. Filter the reaction mixture through a pad of Celite and remove the solvent under reduced pressure. The removed e residue was subjected to furanone dichromatography, 90:10:1:1 -80:20:2: ICHCi, :Me.

Elutes with a gradient of H:H2O:HCO*H, giving the title compound 3 as a white solid. 7.5 mg (yield 64%) was obtained.

Nl/R (CD30D): δ7.03 (2)! , ml, 6.66 (2 H, m), 4.4-4.05 +5H, m).

4.03-3.50 (4H, ml, 3.06-2.54 (21, m), 2.38 (2M, m), 2.14 (2H.

m), 1.96-1.62 (2M, m), 1.4El (9M), 1 ．． 3E+ (3H, m), 1.22 (3M.

(4RS, 5S)-5-(serylalanylalanyl)amino-6-(4-hyalanyl) droxy)phenyl-4-hydroxy-1-oxo-hewi/rubaryl valine hydrochloride C) 5.9 mg (7.7 mmol) of the compound of Example 83(b) was treated with trifluorocarbon Treated with 0.25 ml of acetic acid and stirred the resulting solution for 90 minutes. With methanol One drop of concentrated hydrochloric acid was added at and the mixture was evaporated under reduced pressure. Tritify the residue with Et and O. The title compound was obtained as a white powder.

NMR (CD30D): δ7. Q3 (2H, m), 6.68 (2 also m) , 4.42-4.1745 also mH7tki rainbow d's), 0.98 (12H, ml.

MS (FAB): mlZ 667.4 (M+H) 10.

Example 84 (4R3,5S)-5-(t-butoxycarbonylserylalanylalanyl)a Mino-4-hydroxy-6-(4-hydroxy)phenyl-1-oquin-hexy Production of rubaryl amide a) Example 13 at -40°C under argon atmosphere (c ) to a THF solution of 25 mg (58.5 μmol) of the compound N9.9μ! (90 μmol), then 7.6 ml of inbutyl chloroformate (,58,5 mmol) was added. After stirring for 15 minutes, 14 mg of valinamide (120 mmo I) was added and the resulting mixture was warmed to room temperature and stirred overnight. Ta. The reaction mixture was diluted with 5% HCI and extracted three times with CHCl3. vacuum the solvent and the residue was subjected to flash chromatography with 1:9 MeOH : 23.3 mg of the title compound as a yellow solid (yield 75%) as eluted with CHCL I got it.

NMR (CD30D/CDC13): δ7.36 (5H, m), 7.12 +2H, d), 6.91 +2H.

(4RS, 5S)6-(4-benzyloxy)phenyl-5-(t-butyloxy) cycarbonyl)amino/-4-hydrobyl-cy-1-o+v-hexyl-valylamide b) 1:4 CH=CQt:MeOH(2,5xc)i:: produced above NaB H-C3119, 80 mmol) was added. After stirring for 15 minutes, the mixture was %Hoff and extracted several times with CHtCρ. The combined organic extracts were , O and dried (MgSO,). filter and remove the solvent in vacuo; The title compound (2329, yield 1.0%) was obtained as a white solid.

NMR (CD30D/CDC131: δ7.36 (5H, ml, 7.13 (2H, d), 6.90 +2H.

d), 5.04 (2-rl, 51/4.20 (IH, @i or t), 3.67 +1)! , ml, 3.53fIH, ml, 3.02-2.3 0 (4H, m), 2.10 (IH, rn+, 2.02-1.61 +2 H, rh+.

1.35 (9H, 2-1f&), 0.97 (6H, company d's).

MS (TAB): mlz 528.3 (M+)i)”.

(4R3,5S)-6-(4-benzyloxy)phenyl-5-ami/-4-hypolymer Droxy-1-oquinohexyl-valylamide hydrochloride C) The compound of Example 84(b) (22:9.42 mmol) was added to neat trifluoro Dissolve in vinegar @ (0,1: rL) for 1 day. After 1 hour, methanol was added and then , concentrated HCl2 (2 drops) was added. Remove the solvent in vacuo and mark the hygroscopic solid. Compound (22:, 9) was obtained.

NKL (CD30D): δ7.35 (SH, ml, 7.20 (28 , 2 double All +, 6.9El (2H, 2Q 艷11), 5.07 +2H, 51, 4, 21fIH, ml, 3.83-3.53 (1 formula % formula % ( 2 m) t 2-09 <IM, m) t 1.98-L-62 + 2 formula ml, 0-9 8 (fi)I, m) -(4R3,5S)5-(t-butyloxycarbonyl (ru-rumethyl)serylalanyl-alanyl)amino-6-(4-benzyloxy) ) phenyl-4-hydroxy-1-oxo-hexyl-t<11 amide d) Add t-butyloxycarboxylic acid to THF (shiQ) at -40''C under argon atmosphere. Bonyl-(0-phenylmethyl)serylalanylalanine (20zg, 46 Into a solution containing N-methylmorpholine (8.2 mmol, 75 mmol) ) was added, followed by ionophyl chloroformate (6.0ff12, 46 mmol). ) was added. Obtain 7: Stir the mixture for 15 minutes and add Phosphorus (8.2-C, 75 mmol) was added slowly followed by THF (0. A solution containing the compound of Example 84(c) (22 gg) was added to 8,:C).

The resulting mixture was allowed to warm to room temperature and stirred overnight. - l of the reaction mixture that solidified overnight: I CHCci: Dissolved in MeOH and washed with 5% HC12. Reduced organic extracts Evaporated under pressure. Residue (51+9) 9: 1: 0.5 CHC+2 a:MeOH:Dissolved in acetic acid (21) and applied to a 6”X15xx flat acid column. and eluted with 9: I C H CQs: MeO H to give a yellow solid. (19.3 zg) was obtained. Analytical HP LC (Silica, 9: I C H tc (lx: MeO H mobile phase: detection! 2 7 5 nm) results , the ratio of 4-hydroxy diastereomers is 2:1. 80:20:2 CHCI2. Furanone chromatography on silica using :MeOH:H,0 Further purification was carried out by distillation to give the title compound as a white solid (13.8=t, yield 39%).

NMR (CD30D/CDC131DMSO-d6): δ7.32 (10 3+ml, 7. i2+23! /　m).

6, 82 <2Hz ml, 5.00 +2H, s), 4-19 +54 ”), 4-00 (lh, m) - 3-68 (4H, ml, 3.1 0-2.60 (2B, ml, 2.42 (2H, ml, 2. 13 (1% ml, 1.X9- 1,63 (2)! , rn), 1.50 (9H, S), 1.37 + 3a, *j>d’s)r 1.12+3H, d), 0. 95 (to 6, m).

MS (rAB): mlZ 847.5 CM+H)".

(4RS,5S)5-(t-butyloxycarbonylserylalanylalanyl) Amino-4-hydroxy-6-(4-hydroxy)phenyl-1-oxo-hexyl syl-valylamide e) MeOH (5z) of Example 84(d) prepared above Add 10% para to a solution containing the compound (13,8R9, 16.3 mmol). Dium-activated carbon (7Taku) was added. Bubble hydrogen gas through the resulting mixture; This was maintained at 1 atm H for 24 hours. Filter and remove the solvent under reduced pressure to obtain a white The title compound (10.7r.9, yield 99%) was obtained as a colored solid! =.

NMR (CD30D): δ7.05 (2 to 2, 2 * store), 6.64 (2L 2 = tJ & l,.

2i Hayashi), 0.95 (6H, IT++.

MS (TAB): mlz 667, 2 (M+H) j.

Example 85 (IR,2R)-2-(((Is,2S)-1-hydroxy-2-amine-3- cyclohexyl)propyl)cyclopentanecarbonyl-valylvaline methyl ester Stell hydrochloride (IR,2R)2-(((Is,2S)-1-hydroxy-2-t-butyloxy cyclobencunecarbonyl- Valylvaline methyl ester a) Using the production method of Example 27(a), Example Compound 6(b) (54Q, 150 micromol) was added to N-hydroxybenzot lyazole (40zy, 300 micromol), pcc (33,=y, 170 pm) cyclomol), N-methylmorpholine (40μρ, 380 μcmol) and Reacts with valylvaline methyl ester hydrochloride (8029,300 micromol) The title compound was obtained.

(IR,2R)2-((IS,2S)-1-hydroquine-2-t-butyloxy Cyclobonylamino-3-cyclohexyl)propyl)-cyclobentanecarbo Nirubalylvaline methyl ester b) Example 85 in MeOH (3-:C) Compound (a) (10, 1, 17, 4 mmol) was added. 8 liquid, 60psi pto, (10Kg) and - by Parr (Parr) hydrogenation equipment at gHt. Shake together. After 26 hours, the mixture was soaked in Celite's bud. and evaporated under reduced pressure. The residue was converted to 25' I CHC (b: Purification by flash chromatography eluting with MeOH gave the title compound (9R9, yield 89%) was obtained.

N (CDC13): δ6.58 (IH, d), 6.44 (IH, di , 4.68 (1 type dl, 4.51 + 11. dd), 4.23 (IH , br t), 3.74 (3H, s), 3.60 fIH, rll), 3.47□ (IH, ml, 2.68 (IH, br t), 2.39-2.0 2 (4M, ml, 2.0-1.5 (12HB ml, 1.45 (9H, sl, 1.40-1.08 (6H, ml, 0 ．． 94 (12M, ml.

(IR,2R)-2-(((Is,2S)-1-hydroxy-2-amino-3- cyclohexyl)propyl)-cyclopentanecarbonyl-, Tyl ester/hydrochloride C) Compound of Example 83(b) (92 g, 15.5 mmol) in neat triflu Dissolved in oroacetic acid and stirred for several minutes. The solution was evaporated under reduced pressure and the residue was Dissolve in eOH and treat with concentrated HCC. Removal of solvent in vacuo yields the title compound. Obtained.

Example 86 (IR,2R)2-(((IS,2S)-1-hydroxy-2-(ceryl a ranylalanyl)amino-3-cyclohexyl)propyl)cyclopentanecal Production of Bonyl-valylvaline methyl ester/hydrochloride (IR, 2R) 2-(( (IS, 2S)-1-hydroxy-2-(t-butyloxycarbonyl-(0- phenylmethyl)serylalanylalanyl)amino-3-cyclohexyl)pro pill) cyclopentane carbonyl-valylvaline methyl ester a) Add t-butyl oxychloride to THF (0.5x□) at -40°C under argon Rubonyl-(0-benzyl)serylalanylalanine (6,619,15 mmol Add N-methylmorpholine (2,2xQ, 20 mmol) to a solution containing and then isobutyl chloroformate (2°OxQ, 15 mmol) was added. . The resulting mixture was stirred for 15 minutes and further treated with N-methylmorpholine (2,23! ! 2.20 mmol) and then Example 85 (c ) (15 mmol) was added. Allow the resulting mixture to cool to room temperature. The mixture was warmed to room temperature and stirred overnight. The mixture was partitioned between EtOAc and dilute HCC.

The organic extract was washed with an aqueous solution of N aHCOs, a saturated aqueous solution of NaC4, and dried. (M　g　Oa　). Whitening is achieved by filtering and removing the solvent in vacuo. A colored solid (12m9) was obtained. The product was converted to 25 + I CHC (!.

:Produced by flash chromatography eluting with MeOH to give a colorless glass. The title compound (6.2 gg, yield 46%) was obtained in the form of a stick.

N! -IR (CDC13): δ7.35 (5H, ml, 7.0-6.6 (5H, m), 5.49 (IH, brd), 4.54 (2H, s), 4.52-4.08 (5H, m), 3.99 (IH, br), 3.78 (2H.

m) t 3-12 (3Hp s) t 3-42 (IHt br t)t 2-71 (YHt mL 2.20 @(4Hp m), 1.95-1.55 (12)! , m), 1.46 (9H, s) , 1.40 (6H, d), 1.35-1.06 (6H, m), , 0.9 4 +12H, m).

(IR,2R)2-(((IS,2S)-1-hydroxy-2-(t-butyl) xycarbonylserylalanyl-alanyl)amino-3-cyclohexyl)pro pill) cyclopentane carbonyl-valylvaline methyl ester b) MeOH (Iz) Compound of Example 86(a) (6,20,6,9 mmol) 10% palladium-activated charcoal (5 mg) was added to the solution containing 10% palladium on activated carbon. the reaction The mixture was bubbled with hydrogen gas and then maintained under 1 atmosphere H1 for 6 hours. blend was filtered through a pad of Celite and the solvent was removed in vacuo. The title compound was obtained as a colorless glass (6KIil).

NMR (CDC13/CD30D): δ4.43 (IH, d), 4.38 -4.05 (4H, ml, 3.85 (21, m), 3.73 (3H , s), 3. g2 (IH, ma, 3.45 (l) I, m), 2.6El (hH.

m), 2.30-2.05 (4H, m), 1.9-1.55 (ko, 2H, m), 1.46 (9H, s), 1.40+68. %jL-d's), 1.35 -1.10 +6H, m), 0.94 +128. m).

MS (FAB): m/z 811.5 (M+H)”.

(IR,2R)2-(((Is,2S)-1-hydroxy-2-(serylalani) rualanyl)amino-3-cyclohexyl)propyl)cyclopentanecarbonyl Rubalylvaline methyl ester hydrochloride C) Compound of Example 86(b) (62 g, 7 mmol) was dissolved in a few drops of trifluoroacetic acid. Stirred at room temperature for 40 minutes Afterwards, the solution was diluted with MeOH and li concentrated H(J!/dioxane) was added. The mixture was evaporated under reduced pressure to give the title compound.

NMR (CD30D): δ4.50-4.15 (4H, ml, 3.93 +4H, ml, 3.71 (3H, s).

3.45 (IH, t), 2.65 (IH, ma, 2.29 +I H, ml, 2.2-2.0 (3H, ml.

193-1.53 (12M, ml, 1.s-1,1 (6H, ml, 1.41 (3M, d), 1.35 (314°d), 0.96 (12) (, ml.

MS (2’AB): m/z 7115 (M+H1”.

Example 87 (4S,5S)-5”(methoxycarbonyl-alanylalanyl)amino-6- Phenyl-4-hydroxy=(1-oxo)hexylroot 1'rilvaline (5S)-5(tert-butyloxycarbonyl)amino/-6-phenyl- (1,4-dioxo)hexyl-valylvaline benzyl ester a) at -40°C (5S)-5-(tert-butyloxycarbonyl) in THF (4112) ) Amino-4-oxo-6-phenylhexanoic acid (16Qsg, 0.500ml mol) and N-methylmorpholine (66a (!.

Isobutyl chloroformate (65 μQ, 0.5 0 mmol) was added over 5 minutes. After stirring for an additional 15 minutes, roughly N- Add methylmorpholine (66 μC) and then add to DMF (1,5+y12). Valylvaline benzyl ester hydrochloride (205u, 0.60 mmol) was added. The resulting mixture was slowly warmed to room temperature and stirred for 15 hours. Applicable The mixture was diluted with CHCQs and washed with 10% HCQ, then with H,O. Ta. The organic extracts were evaporated under reduced pressure and the residue was subjected to flash chromatography ( 20:I CHCI2s:MeOH) to give the title compound as a white solid. (253 J!9, yield 85%) was obtained.

(4S, 5S) 5-(tert-butyloxycarbonino L;)amino-4 -Hydroxy-6-phenyl-(1-oxo)hexyl-valylvaline benzyl ester b) The product of Example 87(a) (253x9.0.415 mmol) in MeOH Dissolved in 5M12 and added N aB H4 (17R9, 0.45 mmol) . After stirring for 20 min, the mixture was diluted with 5% HCQ and extracted with CH, CQ, Ta. The solvent was removed and the residue was purified on silica eluting with 2% MeOH in CH,CQ. The title compound purified above by HPLC and eluted first (75.6g9) and then the corresponding (4R) diastereomer (135R9).

(4S,5S)-5-ami7-4-hydroxy-6-phenyl-(1-oxo) Hexyl-valylvaline benzyl ester hydrochloride C) Production of Example 87(b) The sample (75 m9) was dissolved in TFA (0.5 m). After 5 minutes, MeOH (211 2) was added, and then concentrated HC3 (50 uQ) was added. The solution is thickened to a rubbery consistency. Condensed, tritinylated with ether and dried to a white powder (70r9) I got it.

(43,5S)-5-(methobowdicarbonyl-alanylalanyl)amino-4- Hydroxy-6-phenyl-(1-oxo)hexyl-valylvaline benzyle Stell d) Methoxycarbonylalanylalanine (29 , 7xy, 0.136 mmol) and N-methylmorpholine (17Mg, 0. Isobutyl chloroformate (17.6 μQ, 0.1 36 mmol) was added over 5 minutes. After stirring for an additional 15 minutes, roughly - Add methylmorpholine (17 Mg), then 2: ] THF: (4S,5S)-5-amino-4-hydroxy- in DMF (1,5!F12) 6-phenyl-(1-oxo)hexyl-valylvaline benzyl ester/hydrochloric acid A solution of salt (6819,0,124 mmol) was added. the resulting mixture was slowly warmed to room temperature and stirred for 15 hours. Mixture 9:l CHC (! s: diluted with MeOH, washed with 10% HCC, then washed with H2O. Yes The extract was evaporated under reduced pressure to give the title compound (95xy) as a white solid.

TLC (9=l CHCl3:MeOH): Rf-0,5°NMR (CD 30D/CDC13): 57.4-7.25 (10M, m), 5.1 7 (2H, dd), 4.45 (1 mokasu furnace +:), 4.30 (IH , ml, 4.18-4.00 (3H, ml, 3.69 (3H, s).

3.56 (IH,' ml, 2.88 (2H, ddd), 2.32 (2H, mI, 2.19 (IH, ml.

2.03 (1M, ml, 1.72 (2H, m), 1.34 (3H, d), 1.28 (3H, d), 0.91 (12H/ad's) - MS (E’AB): m/z 712.4 (M+H)”.

(4S,5S)-5-(methoxycarbonyl-alanylalanyl)amino-6- Phenyl-4-hydroxy-(1-oxo)hexyl-/sl rilvaline e) Product of Example 87(d) in MeOH (5aR) (85*9.0 10% palladium-activated carbon (13+9) was added to a mixture of Ta. Hydrogen gas was introduced into the reaction mixture, which was then heated under a hydrogen atmosphere for 3.5 Time was maintained. The mixture was filtered through Celite and under reduced pressure The title compound (70°5zy, yield 95%) was obtained as a white solid.

NMR (CD30D): δ7.23 (5H, ml, 4.37-4.00 (5H, ml, 3.66 (31(, s).

3.58 (IH, ml, 2.88 (2H, ddcl), 2.32 +2 H, m), 2.18 (IH, mI.

2.06 (l)! , m), 1.73 (2H, ml, 1.32 (3H , d), 1.26 (3H, d), 0.95 (121, m).

MS (r): m/z 622.3 (M+H)”.

Example 88 (4S,5S)-5-(carbobenzyloxy-alanylalanyl)amino-6 -Production of phenyl-4-hydroxy-(1-oxo)hexyl-valylvalinol Construction The title compound was obtained in the same manner as above.

NMR: (CD30D/CDC13): δ7.2-7. -0 (log, ml, 4.93 + 2H, di, 4.1 (IH, meeting t74'l: L4.00 -3.80+3M,'rn), 3.50-3.35<4H,ml, 2.68(2 ! ’,.

Example 89 (4S,5S)-5-(4S,5S)5-(carbobenzyloxyalanylara Nyl)-amino-4-hydroxy-6-phenyl-(1-oxo)hexylva Phosphorus isobutyramide (5S) 5-(tert-butyloxycarbonyl) Amino 4-oxo 6-phenylhexanoic acid benzyl ester a) Dry acetic acid (5S)-5-(tert-butyloxycal) to tonitrile (10zi2) (bonyl)amino-4-oxo-6-phenylhexanoic acid (1 mmol) was added. To the solution, with stirring, was added DBtJ (1 mmol), followed by benzyl bromide. (1.5 mmol) was added. After 2 hours, the mixture was diluted with 5% HC (CH Extracted with tCh. The organic layer was concentrated, and the residue was subjected to furasochinichromatography ( Purification by CHCff, ) gave the ester (370o).

NMR (CDC13): δ7.4-7.0 (log, ml, 5.10 ( 2L sL 5.03 (LH, d).

4-51 (IH, q>t 3-18-2.83 (2J!-m), 2- 13 (2H + ml, 2-60 (2H, m>B (4S, 5S) 5 (tert-butyloxycarbonyl)amino- Benzyl 4-(tert-butyldimethylsiloxy)-6-phenylhexanoate ester b) Preparation of Example 89(a) in MeOH according to the preparation method of Example 87(b). The product was reduced with NaBHa (96-99% yield). The phase of the alcohol obtained The astereomeric mixture was dissolved in DMF at a concentration of approximately 2M and imidazole (2,4 ff1) and tert-butyldimethylsilyl chloride (1,2 equivalents) were added. Ta. The mixture was stirred for 2 days, then diluted with water and extracted with ethyl acetate. organic The layers were dried (MgSO,) and concentrated. Flash chromatography of residue (5:l hexane:EtOAc), then HPLC (15:1 Purified with hexane: E to A c) and the title compound eluted first (harvested) 25%) and then the corresponding (4R)-siloxybenzyl ester 70 %) was obtained.

(4S)-isomer: NFm (CDC13): δ7.4-7.12 (10H, m), 5.0 8 (2H, sl, 4.67 (IH.

d), 3.85 (IH, qL 3.68 + IH/m), 2.72 ( 2H, m), 2.36 (2H, ml.

17B (2H, m), 1.41 (9H, s), 0.88 (9H, s) , 0.10 (6M, d).

MS’ (E’λB): mHz 529 (M+H)”.

(43,5S)-5(tert-butyloxycarbonyl)ami/-4-(te rt-butyldimethylsiloxy)-6-phenylhexanoic acid C) Example 87 (e ) The product of Example 89(c) was hydrocracked to form a brittle foam. The title compound (yield 99%) was obtained.

NMR (CDC13): δ7.35-7.10 (5H, ml, 4.69 (2H, br d), 3.88 (1 ratio (11/3.70 (IH, ml, 2.72 +2H, ml, 2.31 +28. m), 1.78 (28 , ml.

1.30 (9H, s), O, B9 ζ9J (/ Sat CL Yu O (6H, d).

MS (E’AB): mHz 43B, 2 (M+Fri).

(4S,5S)-5-(carbobenzyloxyalanylalanyl)ami/-4 -(tert-butyldimethylsiloxy)-6-phenyl-(1-oxo)hex Syl-valine isobutyramide d) Performed according to the method for preparing Example 87(a) Bling the product of Example 89'(C) with valine isobutyramide hydrochloride. and crude (4S,3S)-5(tert-butyloxycarbonyl)amino-4- (tert~butyldimethylsiloxane)-6-phenyl Sobutyramide was obtained (80% yield) and was prepared according to the preparation method of Example 87(c). This was directly converted to the corresponding hydrochloride. Next, m-hydrochloride was prepared in the same manner as described above. When combined with bobenzyloxyalanine, the compound 4yA becomes a white solid. was obtained (yield 60%).

DC13) for NMR: δ7.45-'7.10 (10H, ml, 6.38 0. )! , ml, 5.08 (2H.

ml/　4.21　+21. rn), 3.70 (1) I, m), 2. 73 (2Hr m), 2.33 (IH/m) 12.12 (2H, m) , 1.25 (4H, ml, 0.91 (22H, ml, 0.10 (6H , d).

MS (rABl: mlZ 697.3 (M+H)".

(4S,5S)-5-(carbobenzyloxyalanylalanyl)amino-4- Hydroxy-6-phenyl-(1-oxo)hekyneluvaline isobutyramide e) Tetrabutylammonium bromide was added to the product of Example 89(d) in THF. (5 equivalents) was added. After stirring for 14 hours, the mixture was diluted with water and dichloromethane. Extracted with lomethane. Removal of the solvent gave the title compound as a white solid.

NMR (CDC13/CD30D): δ7.2B-7.05 (IOH, ml , 4.96 (2H, 5), 4.02-3.76 (4H, ml, 3.4 7 (IH, ml, 2.9-2.60 (4H, ml, 2.13 (2F, , ml.

1.82 (11 (, q), 1.55 (34 ml, 1.09 (4! (, d) ), 0.70 +12H, d).

MS (r): mHz 583.3 (M+H)".

Example 90 (4RS, 5S)-5((tert-butyloxycarbonyl)-isoeisyl ) Amino/-7-methyl-4-hydroxy-1-oxo-octyl-leucyl-a Production of spartic acid methyl ester (3S) -5-(tert-butyloxychloride) (cycarbonyl)amino-7-methyl-4-oxo-octanoic acid a) Bo for Boc-phenylalanine N-methoxy-N-methylamide c-Phenylleucine N-methoxy-N-, except that methylamide was used. Following the preparation method of Example 12(a,b), the title compound was obtained.

NMQ (CDC13): δ5.0 fIH, d), 4.35 fIH, m l, 3.5 (1 yt 53/2°8 (2H, ml, 2.6 (3H, t ), 15 (9H, s), 0.90 (8M, d).

(5S) 5 (tert-butyloxycarbonyl)amino-7-methyl -(1,4)-dioxo-octyl-leucyl-(O-benzyl)asparagine acid methyl ester b) The product of Example 90(a) and leucyl-(0-benzyl)asparagine Coupling method of Example 34 (a) using acid methyl ester/hydrochloride The title compound was obtained according to the following procedure.

(4R3,5S)-5(tert-butyloxycarbonyl)amino-7-methy -4-hydroxy-1-oxo-octyl-leucyl-(0-benzyl)as Partic acid methyl ester C) The product of Example 90(b) was dissolved in excess of After reduction with NaBH,CN (2 days), water treatment and furanosilachromatography. After treatment, a II mixture of 4R and 4S diastereomers of the title compound was obtained. To.

NMR (CDC13): δ7.35 (5M, sl, 7.05 (I H, d), 6.15 (1:i, ml.

5.15 (2H, sl, 4.85 (2H, ml, 4.55 (IH, d ), 4.4 (IH, ril, 3.65 (3H, s), 2.9 (2H, m), 2.4 (2, t), 1.5 (9Ht s), 0.90 ( 12H.

ml.

MS (FAB): 622.3 fM+H)+・(4RS, 5S) 3-( (tert-butyloxycarbonyl)ynleucyl)amino-7-methyl-4 -Hydroxy-1-oxo-octyl-leucyl-(0-benzyl)asparagi d) Using Boa-inleucine instead of Cbz-alanine Example 9 was prepared according to the manufacturing method of Examples 76(b) and 77(a), except that The title compound was obtained from the product of 0(c).

NKR (CDC13): δ7.4 (sl4.sL 7.0 +IH, d) , 6.3 (0,5H, d), 6.2 (0,5H, d), 5.15 (2 H, s), 5.05 (2H, ml, 4.9 + 11. m), 4.5 +iH.

ml, 3.7 +3H, d), 3.0 (2H, ml, 2.45 +3H , ml, 1.5 (15B, s), 1.3+158. s), 0.90 ! 19H, m1.　　　.

MS (rAB): 735.5 (M+H)”.

(4R3,5S) 5 ((tert-butyloxycarbonyl)inroy sil)ami/-7-methyl-4-hydroxy-1-oxo-octyl-loinyl - Aspartic acid methyl ester e) Practical according to the production method of Example 77(b) The title compound was obtained from the product of Example 90(d).

N calm: δ4.55 (IL t), 4.35 (IL I'11) / 3.8 (2H, ml, 3.65 +3H, 51/3.45' (IH, ml, 2. 7 (IH, rn), 2.5 (1) i, m), 2.3 (2H, m), 14 (9)i.

S) + 1.2 +6H, sL 0-90 (15Hr m) -MS (F stone ): 667.4 (M+Na)”; 643.5 (M-:’ri-・real Example 91 (4R,5S) and C''(4S,,5R)-carbobenzyloxy-alanyl- (3-hydroxy-5-amino-6-phenyl)hexanoyl-valyl-(4- Production of Boc-valyl-(4-aminomethyl)pyridine &) CHICI2t 100Jl+! +: 4-7 minnow methyl viridi 7(,2 , 59,23 mmol) together with Boc-L-valine (5,09,23 mmol). ) was dissolved. Dicyclohexylcarbodiimide (4,759,23 mmol) was added and the reaction was stirred overnight. The reaction mixture was filtered and diluted with 1N NaHCOs ( 3X50P! I2”), washed with water (1x50+f2) and dried with Na, SO2. I set it. The solvent was removed under reduced pressure and the residue was tritinilated with hexane and filtered. and crystallized. The partially purified product was purified on silica gel in ethyl acetate. Purified by passing through a bed. Removal of solvent yields white solid 3.64 g of product was obtained.

Nx? t (CDC13): δ 0.83 (d), 0.90 (6H, dl, 1.30 (9H, sl, 3.8|4.0 (IH, dd) 4.38 (2H, di, 5.2 +18. d ), 7.1 (d), 8.5 (4H, a+, (5S)-carbobe Oxygenyloxy-alanyl-(4-oxo-5-amino-6-phenyl) Irubalyl-(4-aminomethyl)pyridine b) Boa-L-valyl-(4- Aminomethyl)pyridine (2,Os+) was treated, evaporated to dryness, and the residue was dissolved in water. Dissolve, filter and lyophilize L-valyl-(4-aminomethyl)pyridine. ・Dihydrochloride was obtained.

L-valyl-(4-aminomethyl)pyridine dihydrochloric acid [(0,169,0,5 mi Limol) and Boc-4-oxo-5(s)-ami7-6-phenyl The acid was dissolved in DMFIO; J2. To this, benzotriazol-1-ylol oxytris(dimethylamino)phosphonium hexafluorophosphate (0, 3329,0,75 mmol), 1-hydroxybenzotriazole (0,10 1y, 0.73 mmol) and diisopropylethylamine (0,0979 , 0.75 mmol) was added and the reaction mixture was stirred at room temperature overnight. Saronibe Oxytriazol-1-yloxytris(dimethylamino)phosphonium to cattle Fluorophosphate (0,332y, 0.75 mmol), 1-hydroxy Benzotriazole (0,1019,0,75 mmol) and diisopropyl Add ethylamine (0,0979,0575 mmol) and add another 24 min at room temperature. time, the reaction proceeded. After evaporating the solvent, the reaction mixture was dissolved in ethyl acetate, Washed with 1o% KtCOs, saturated NaHSO, and water, dried with NatsOa. Dry and concentrate to an oil. Silica using 5% methanol/methylene chloride Purify on gel to obtain (4S)Boc (3-oxo-4-amino-5-phenylene) m)hexanoyl-valyl-(4-amino/methyl)pyridine 99R9 was obtained.

5(S)Boc-(4-oxo-5-amino-6-phenyl)hexa obtained above Noyl-valyl-(4-aminomethyl)pyridine was treated with 4NHC+ for 31 hours at room temperature. 2/treated with dioxane, evaporated to dryness, dissolved the residue in water, filtered and lyophilized. After drying, (,5S)3-oxo-5-amino-6-phenyl)hexanoyl- Valyl-(4-aminomethyl)pyridine dihydrochloride 70g9 was obtained.

In DMF, carbobenzyloxy-thi-alanine (51F, 9.0.23 mmol) ), benzotriazol-1-yloxytris(dimethylamino)phosphoni umhexafluorophosphate (102 = 9.0.23 mmol), 1-hydro Roxybenzotriazole (31+9.0.23 mmol) and diisopropylene (5S)3-oxo with ruethylamine (30r,y, 0623 mmol) -5-amino-6-phenyl)hexanoyl-valyl-(4-aminomethyl)pi Dissolve lysine dihydrochloride (67 Tekken, 0.15 mmol! J mole) and stir the reaction at room temperature. Stir overnight. The reaction mixture was evaporated, dissolved in acetonitrile and dissolved in 4xQZ min. 10% acetonitrile-water-0,1% TFA to 50% acetonitrile over 30 minutes Beckman Ultras using a gradient of tolyl-water-〇, 1% TFA. HPL for preparation on Beckman Ultrasphere ODS Purified by C. Appropriate fractions were pooled, evaporated to dryness, and lyophilized from 1% acetic acid. Drying yielded the desired product 29x9.

FAB-MS m/z 616.2 (M+H)", 614.2 (M-H)- 0(3R,4S) and (3S,4R)-carbobenzyloxy-alanyl-( 3-Hydroxy-4-amino-5-phenyl)hexanoyl-valyl-(4-a (minomethyl)pyridine c) (4S)carbobenzyloxy-alanyl-(3-o quin-4-amino-5-phenyl)hexanoyl-valyl-(4-aminemethyl )-pyridine (14X20t9.0.033 mmol) dissolved in methanol 5λQ and sodium borohydride (1,4ty, 0.036 mmol) was added.

After 30 minutes at room temperature, add more sodium borohydride la9 and add another 3 minutes at room temperature. The reaction proceeded for 0 minutes. The solvent was removed by cyclization and the residue was dissolved in acetic acid and frozen. Dry. Using 28.5% acetonitrile-71.5% water-〇, 1% TFA Beckman Ultrasphere ”) Diastereomeric mixtures of alcohols were determined by preparative HPLC on OD The compound was separated.

Appropriate fractions were pooled, evaporated to dryness and lyophilized from 1% acetic acid to give isomer A 29, and isomer B was obtained as 6Fi.

Isomer A: FAB-MS m/z 618.5 (M-', H)', 616.4 (M-H) -0 HPLC: k' = 9.6 (10% acetonitrile-water over 30 minutes) -0.1% TFA to 5o% acetonitrile-water-Q, 1% TFA gradient Using the Beckman Ultrasphere here”)　　○DS) Oral isomer B: FAB-MS m/z 618.5 (M-i-H)', 616.4 (M-H )-0 HPLC: k' = 9.9 (10% acetonitrile- Water-0,1% TFA to 50% acetonitrile-water-0,1% T　F, A graph Beckman Ultrasphere (Beclvan U 1t) rasphere”) OD S).

Example 92 Production of (3S)-Boc-valyl-4-methyl-3-aminopentan-2-one Boa-L-valine N,O-dimethylamide a) dimethylhydroxylamine Hydrochloride (3,91 g, 40 mmol) was dissolved in CHtCQt25prQ and heated to 0° Cooled to C. Add triethylamine (4.14 g, 40 mmol) and concentrate A thick suspension was obtained which was warmed to room temperature.

Boa-L-valine (8,689,40 mmol) was added to THF5Qx (l and c It was dissolved in H, Cct 180 yen and cooled to -20°C. N-methylpiperidine ( 4,88+ff, 40 mmol) and then ethyl chloroformate (4,3 49.40 mmol) was added. After 5 minutes, add dimethylhydroxylamine solution. After 3 hours of addition and warming of the reaction to room temperature, the reaction mixture was heated to IN HC (!(2 Wash with X100z□, IN Na0H (2X100!, and dry with Na and SO2) The product was purified and evaporated to a colorless oil, which was used without further purification.

3(S)-4-methyl-3-t-butyloxycarbonylaminobencune- 2-on b) Boa-L-valine N10-dimethylamide (2,6y, 10 mmol) diethyl ether 30 = (methyl bromide dissolved in diethyl ether) A 3M solution of nesium was added dropwise to IL:((51 mmol).The reaction mixture was kept at room temperature. Stir for 3 hours, then pour into ice water and add 1N Hci! Acidify to pH 2 with Extracted with ethyl acid (3×50×□). The extract was dried with NatSO, and the layer 1. A colorless oil was obtained, which was purified on silica using 2% methanol/CH,CQ2. Purification by gel chromatography gave the desired product 1.749.

NMR (CDC13): δ0.82 (3H, d), 1.05 (3Gl, d), 1.45 (9H, s), 2.2 (3H, s) 4.3 fIH, ml, 5.25 (LH, br, d).

(3S)-Boa-valyl-4-methyl-3-ami/pentan-2-one C) 4-methyl-3(S)-t-butyloxycarbonylamine for 30 minutes at room temperature. Nobentan-2-one was stirred with 4N HCQ/dioxanthate and concentrated under high vacuum. The remaining HCC was removed by condensing and then evaporating CH, Cj2 a number of times.

The obtained hydrochloride was dissolved in CH, CC, t and neutralized with 1 equivalent of triethylamine. Ta. Then Boc-L-valine (1 equivalent) and dicyclohexylcarbodiyl (1 eq.) was added and the reaction was stirred at room temperature overnight. Filter the reaction mixture , IN　N　a　2　washed with COy and dried with NatSO, 1. Layer line, The dipeptide was obtained and then purified by silica gel chromatography. Manufactured.

(4S,5S)-benzyloxycarbonylalanyl-alanyl-(6-phenylalanyl) -5-amino-4-hydroxy-hexanoyl)-valyl-(4-methyl-3 (S)-aminopentan-2-one) d) of Example 92(c) for 30 minutes at room temperature. The compound was treated with 4N HCI2/dioxane, concentrated under high vacuum, cH,CQt The remaining HCl2 was removed by evaporation several times from (3S)valyl-4-methyl -3-aminopentan-2-one hydrochloride was obtained. In the manufacturing method of Example 89 The title compound was obtained by replacing valine isobutylamine with this compound.

Example 93 (O3) Boa-5-ami/-6-phenyl-trans-hex-3-enoic acid manufacturing Monomethyl trans-3-hexenoic acid a) diazald (D1azald”) (21,49,100 mmol) was dissolved in ether and heated at 65°C with KOH (5 9), water (8jlQ) and ether (25x□). Obtained The diazomethane obtained was directly mixed with β-trans-hydromuconic acid (10 g, 69 mmol). molar) solution. The solution was evaporated and the residue was diluted with 2% acetic acid/chloroform. purified by flash chromatography on silica gel using 1.249 of the desired product was obtained.

N! 'R (CDC Yu3): δ 10.85 (IH, s), 5.75 (2H, m), 3.68 +3H, sL3.23 (4H, m).

6-(2-oxo-4-(phenylmethyl)-3-oxazolidinyl)-tran (4S)-methyl su-hex-3-enoic acid b) THF 5QxQ to')xf monomethyl tran with h7mine (l, 42; 7j, 10.2 mmol) Dissolve su-3-hexenoic acid (1.24 g, 7.8 mol) and cool to -78°C. did. Pivaloyl chloride (1°O62 g, 8.6 mmol) was added and the reaction was quenched. Stir at 78°C for 15 minutes, then warm to 0°C for 45 minutes 1::THF 302i2e(S)-benzyloxazolidinone (2,59,14゜1 mmol ) was dissolved and cooled to -78°C under argon. Butyl lithium (hexane) 2.5M of 5.6=Q> was added slowly and the mixture was stirred for 5 minutes. The mixed anhydride obtained above was cooled to -7860, and the lithiated oxazolidinone was Added via eule.

The reaction was stirred at -78°C for 15 minutes, then warmed to room temperature for 1.5 hours. anti The reaction mixture was quenched with IM NaH3O4100112 and the THF was removed under reduced pressure.

The aqueous residue was extracted with CH,C4), and the combined extracts were dissolved in 0.25M NaH3 Washed with O4, dried with 1g5OJ and evaporated to dryness.The crude product was By furanosinichromatography on silica gel in 0% ethyl acetate/hexane. Purification yielded the desired product 1.409.

FAB-MS, m/2318 (M+H)”.

6-(2-oxo-4-(phenylmethyl)-3-oxazolidinyl)-5-( phenylmethyl)-trans-hex-3-ene a (4S.

5S)-methyl C) The oxacylidinone (3.9 mmol) obtained from Example 93(b) was dissolved in THF. It was dissolved in 5QxQ and cooled to -78°C. Potassium hexamethyldisilazi 0.3 M in CTHF, sic, 3.9 mmol) and the mixture The mixture was stirred at -78°C for 15 minutes. Benzyl bromide (0.93m (!, 7.8m) The mixture was allowed to warm to room temperature and stirred overnight. Concentrate the reaction mixture , the residue was purified by gravity chromatography using 20% ethyl acetate/hexanes. to give 0.389 of the desired product (and 0.099 of the 5(R) isomer). Ta.

(3H, ml 4.05 + 2H, ml, 3.60 + 38. sl, 3. 0 (78, ml.

EAB-MS: m/z 408 (M+H).

5-(phenylmethyl)-trans-hex-3-ene F ((5S)-monomethyl le d) THF 13.9=C and water 4.6 JT (in Example 93(c)) RQ oxazolidinone (0,389,0,93 mmol) until O″C Cooled. Hydrogen peroxide (0.63:C of a 30% solution) is added, followed by hydroxyl Lithium chloride (78.1 Mg, 2 eq.) was added. After stirring for 30 min at o'C , 1.5M NatS Os, 4.5M (l) was added to quench the reaction. Below, THF was removed, the pH of the aqueous solution was adjusted to 10 with NaHCOs, and acetic acid was added. Washed with ethyl and ether.

The pH was then lowered to 1 by IMHC (and extracted with ethyl acetate. The extract was The combination was dried over N a I S O 4 and evaporated to give the crude product. 2 % Acetic Acid: Fluorized on silica gel using chlorine.

Purification by chromatography yields the desired product 50. ．． I got a 9.

NMR (CDC13): δ9.43 (11,s), 7.25 (SH,m l, 5.64 (2H, ml, 3.65 (3H, s), 3.05 (5H , m).

(5S)-Monomethyl 3-(phenylmethyl)-trans-hex-3-ene Mido e) The carbonic acid of Example 93(d) (50=9.0.2 mmol) was dissolved in toluene 3 Dissolve L in 2C, cool to -0"C. Triethylamine (28, :Q, 0 ．． 2 mmol) and then ethyl chloroformate (166C30, 2 mmol) was added. ) was added. After stirring for 1 hour at −5°C, ammonia was bubbled through the solution. , then warmed to room temperature and stirred for 2 days. The solution was evaporated and the residue was dissolved in acetic acid. Dissolved in ethyl, washed with 0.5 MXaHC○, and dried over MgSO4. Applicable Evaporation of the solvent gave the desired product 5029.

NMR (CDC13): δ7.25 (5H, ml, 5.84 +2H, m l, 3.6843H, s), 3.0 (5H, ff1l.

5-Amino-6-phenyl-trans-hex-3-enoic acid (5S)-methyl f) Dissolve the carboxydiamide of Example 93(e) in acetonitrile-water (4:l). 1,1-bis(trifluoroacetoxy)iodosobenzene (1,5 equivalents) ) was added and the reaction was stirred at room temperature until completion. The reaction mixture was diluted with 1N Hce and washed with ether. Then, the pH of the aqueous solution was adjusted to 1N NaO The product was extracted with ethyl acetate and the extract was diluted with Na2. Drying with S04 and evaporation gave the desired product.

5-t-butyloxydicarbonylamino-6-phenyl-trans-hex-3 -(5S)-methyl enoic acidg) in DMF with triethylamine (3 equivalents) The compound of Example 93(f) was dissolved. Di-t-butyl dicarbonate (3 units jl ) was added and the reaction was allowed to proceed overnight at room temperature. Remove the solvent under reduced pressure and silify the product. Purified by furanosinichromatography on Kagel.

(5S) -Boc-5-amino-6-phenyl-trans-hexer-3-e acid h) Example 9 to 1:1 dioxane:IN NaOH (1 per fiNaOH) Compound 3(g) was dissolved and stirred at room temperature until the reaction was complete.

The solution was acidified with IN HC (!) and extracted with ethyl acetate to give the desired product. .

Example 94 (5S)-alanyl-alanyl-(5-ami/-5-phenyl-trans-hex Preparation of ter-3-enoyl)-valyl-valinamide According to the conventional method of Example 1, Protective peptidyl resin (5S) BOC-Ala-Ala-(5-amino-6-peptidyl resin) phenyl-trans-hexer-3-enoyl) -Val -Val -B HA was manufactured. Treated with anhydrous liquid HF 10ff (/anisole 1XQ) for 1 hour at 0°C. The peptidyl resin was cleaved and deprotected by. HF under vacuum After removal, the resin was washed with diethyl ether, air dried and treated with glacial acetic acid 2X50x. 12 and lyophilized to obtain crude peptide. 10% vinegar as eluent Gel on Sephadex G-15 using an acid aqueous solution. The crude product was purified by filtration.

Example 95 Production of Asn-Tyr-Pro-Val-Val-NH, 0.5 mmol scale Protected peptidyl resin Boa-Asn-Tyr(BrZ)-Pr in the usual manner o-Vat-\'al-BHAf-manufactured. 1 hour at 0°C, anhydrous liquid HF After removing the cHF that had cleaved the peptide from the resin, the resin was washed with ether. , air dried and extracted with 10% HOA C3X30+Tg. HOA C extract in water and lyophilized to give crude product 145vy. The crude peptide (100 z9) in 1% HOAc, 2.6 x 70c: Sephadex (S ephadex”) G-15 column. Appropriate fractions were pooled and lyophilized. The purified product was 84.4F! I got a 9.

HPLC: ni' = 2.93 ("Milton PRP -1 (Hamilton P RP-IR), 5% CH, CN 10.1% T F A ~ 40% CH, CN/ Q.

1% TFA, 15 minutes).

MS (FAB): m/Z 590 (MH)".

Example 96 Production of Ala-Asn-Tyr-Pro-Val-Val-NH2 Same as above The title compound was obtained.

HPLC: k' = 2.47 (Hamilton P RP -1 (Hamilto n　P　RP-1　5%CH,CN10.1%T　F　A~40%CH,CN 10°1% TFA, 15 minutes).

λ1s (FAB): m/z 661 (M-"H)".

Example 97 S er - G In - A sn - T yr - P ro - V al - Preparation of Val The title compound was obtained in the same manner as above.

HPLC: k' = 2.86 ("Milton PRP -1 (Hamilton PRP-1”), 1O%CH, (, N10.1%TFA~40%CH,CN 10.1% TFA, 15 minutes).

MS (FAB): m/z 806 (M÷H)°.

Example 98 Preparation of Set-Gin-Asn-Cha-Pro-Val-Val-NH2 The title compound was obtained in the same manner as above.

) IPLc: k'=3.31 (Hamilton P RP -1 (Hamilt on P RP-I R), 0% CH3c X10.19’6T F A~4 0% CHsCX10.

1% TFA for 15 minutes).

\TS (FAB): m/z 793 (M-"H)".

Amino acid analysis: Aspl, 0O1Ser0.72, Glui, 02, Pr.

0.97, Val2.01゜ Example 99 , Ac-3er-Gln-Asn-Cha-Pro-Val-Val-NH, Preparation The title compound was obtained in the same manner as described above.

HPLC: k' = 3.27 ("Milton PRP -1 (Hamilton PRP-1”), 5%cHscN10.1%TFA~40%cH,cNlo .

1% TFA, 15 minutes).

MS (FAB): m/z 837 (λi?H)".

Amino acid analysis + Asp 1.00. Ser　O,75,GIuo,99,Pr.

0.96, ~'al　1.90゜ Example 100 Ac-3e r-G I n-As n-Ty r-P ro-Va 1- Production of NHt The title compound was obtained in the same manner as above.

MS (FAB): ! Tl/Z 74B, 7 (M+H)”HPLC: k' -2,08 (Hamilton PRP-1", 5% C H3CN10. IS TTA to 40r CH3CN10.1% TEA, 15 min,).

Am1no Ac1d Analysis: Input sp 1.00.　 Ser O, 55, Glu O, 98, Pro 1D19゜ Val　O,95,Tyr　O,99゜Example 101 Ac-Asp-Gin-Asn-Tyr-Pro-Va l-Arg-NH, manufacturing The title compound was obtained in the same manner as above.

HPLC: k’-2,04 (Hamilton PRP-10,9CH3 CN/0.1STr included -40% CH3CN/0.1% TE'A, 1 5 min,) MS (FAB): mHz 932 (M+H)”7’ tノ＠ld’k　　　　　　　　　In sp　　1.88. Glu　O,90,E 'ro　O,76, Val　1.00゜Tyr　O,33, company g　101. ・Example 1.02 Production of Asp-Gin-Asn-Tyr-Pro-~'al-\'al-NH, The title compound 1 was obtained in the same manner as above.

? ,? LC: k’-2,44 (Hamilton PRP-10) 5% CH3CN/　O, 1% TFA ~<0% CH3CN/　O11%TTA, 15 min, IMS (E’AB): mHz 833 (M+H1 ”r=) Revised/*”F4〒〒　　　　　　:　　　　　　sp　　2.00. Glu O, 98, Pro O, 96, Val 2.04° Example 103 Ac-Ala-Ala-Set-Gin--Asrl-Tyr-Pr.

Production of Va1-Va1NHt The title compound was obtained in the same manner as above. .

Example 104 Production of Ac-Ala-Gin-Gly-Tyr-Pro-Val-Val-NH. Construction The title compound was obtained in the same manner as above.

HPL (:= k' -4,48' (Beckman t71trasphe re■ODS, 5% C)! 3CN/　O,]JTFλ　-40%　CH3C N/O, 1% TTA, 15 min,) Example 105 D-3e r-Gin-Asn-Tyr-Pro-Va 1-Va l-NH ,Manufacturing of The title compound was obtained in the same manner as above.

TrA 6740% CH3CN/ 0.1% TE”A, 15 wi n,)MS　1rAB):　mHz　805　(M+H)"7mi)7@plasma*1 7　　　　、　　　sp　1.00.　Glu　　O,99,Pro　 1.11. VaL L, s9゜Tyr O, 57, Ser O, 68゜ Example 106 Ac-D-5e r-G I y-, As n-Ty r-P r o-Va Production of l-VaI-XH, In the same manner as above, (Gyuhi compound was obtained.

HPLC: k’-4,16 (Beckrnan Ultrasphere "ODS, 9C, '13CN/O, 1STr included Hano 40% CH3 CN/ O, 1% TrA, 15 min,) MS ffAB): mHz 847 (M+H)”7 Mi no m friend’;?’Xr: Enter sp 1.00. Ser O, 67, Glu O, 99, Pro O,! 93K Val　2.00. Tyr O, 30° Example 107 Production of Ser-Gin-Asn-Tyr-Pro-Ala-\°a1-NHt The title compound was obtained in the same manner as above.

HPLC:　　　k’　　-3,96(Beckmanυ1traspera@ ODS, 5% CH3CN/O,1STr included, 40% CH3CN/ O, i% TTA, 15 win,) MS (F): mHz 777 (841>”Ryo Mino vt*t: Enter sp 1.05.Se r　O,91,Glu　1.00. Input 1a 1.02゜Pro O, B6 ．． Val 1.09. Tyr O, 61° Example 108 Production of Ac-ser-gin-asn-tyr-108Ac-ser-1~NH, Construction The title compound was obtained in the same manner as above.

HPLC: k’-3,87 (Beckman Ultrasphere 0 005.54 CH3CN/0.1STr included A/40% CH3C N/0.1% τrA, 15 min,) Example 1○9 Ac-3er-Gin-Asn-Tyr-Pro-~731Val-A r g -Manufacture of NHt The title compound was obtained in the same manner as above.

Example 110 Ac-Se r-Gln-Asn-Tyr-Pro-Glu-Va 1-xH, Manufacturing of The title compound was obtained in the same manner as above.

HPLC: k' # 3.83 (Beckman Ultrasphe: e■00s, 5% CH3CN/O,lt Example 111 Production of Ser-Gln-Asn-Tyr-Pro-Glu-\'a1-XH1 The title compound was obtained in the same manner as above.

! ! PLC: k’ = 3.35 (Beckman tn sezasphe re■ODS, 5% CH3CN/0.14? TA ~40% CH3C N/　　　0.1%　TFA,　15　win,)　’MS　　(E’λB ): m/z 835 (M 10H)” Example 112 Ac-5e　r　-G　n　-G　y-Ty　r-P　ro-Ly　5 Production of -Va 1-NH, The title compound was obtained in the same manner as above.

Example 113 Production of AC-Se t-Gl n-Asn-Tu r-Pro-NH: Same as above The title compound was obtained in the same manner.

Example 114 Ac-3e r -G l n-Asn-Ty r-P ro-VaI -Va Production of 1-Gln-Asn-NHy The title compound was obtained in the same manner as above.

Example 115 Ac-3e r-Gln-Asn-Phe-Pro-Va I-Va 1-N Manufacture of H. The title compound f: was obtained in the same manner as above.

MS (rAB): m/z 831 (M+H)” HPLC: k’ -3,02 (Hamilton PRP-1', 9'CH3CN1 0.1% T! "A ~, 40■ C”3CN10.1%τrλ, 15 min,).

Example 124 9oc-L-valine Production of inpropylamine Boc-L-paris/(0.5g , 2.3 mmol inpropylamine (0.36 ml 1.4.6 mmol) , Bensito IJ 7zol-1-yloxytowas(dimethylamino)-phosph Onium hexafluorophosphate (2.03g, 4.6mmol), I -Hydrobovine dibenzotriazole (0.62 g, 4.6 mmo I) and di- DMF (,2 5 mI). The reaction solution was stirred at room temperature for 2 hours, evaporated to dryness, and the residue was dissolved in vinegar. Dissolved in ethyl acid, lN N a HSO4, iN NaHCOz and Rinse with water and dry fill with 2 sOa of Na. Remove the solvent under reduced pressure and remove the crude 0.53 g of product was obtained. The product was transferred onto silica gel in 25% 5ieOH/CHCl. , to obtain 358 mg of purified product. Ta.

MS (DC engineering), m/z 259 (M+H)” NMR (CDC13 ): δ0.90 ppm, d, 0.98 ppm d, 6H; 1.16 PPm1 d, 6 towns 1.35 ppm, s, 9H; 4.1 ppm, dd, IM; 5.25 ppm, d, 11 (; 6.14 ppm.

d, IH.

Example 117 Production of Ac-Tyr-Pro-＼’al　Val　:’;Ht Same as above The title compound was obtained.

HPLC: k’-2,96 (Hamilton PRP-10,9CH3 CN/O, 111τy, qν4CnCH3CN/0.1%τF’A, 15 　min’,).

MS (FAB): m/z 518 (M+H) 10 Example 118 Ac-Arg-Se r-G I n-Asn-Ty r-P ro-NHt Manufacturing of The title compound was obtained in the same manner as above.

85 (E’AB): m/z 876 (M+g)” Example 119 Preparation of Ac-3er-Gln-Asn-Tyr-Pro In the same manner as above, the title The compound was obtained. This compound was used in the following examples without further purification.

Example 120 Ac-Se r -G l n-, Asn-Ty r-Pro-Va l -H Manufacture of N iPr The title compound was obtained in the same manner as previously described.

HPLC: ) c’ wm 4.97 (Beckr+an υ1 t=asphe:e■ 005. 5’l CH3CN/@ O, Co τr included ^I 404 C: (3CN/0.1%?TA, 15 min,) MS (TAB): m/z 790.5 (M+H)” 4 members moxibustion fi Ding: Asp 1. OOt Ser O , 72, Glu 1.02.　Pro　　1. I3# Val　O,65,Tyr　O,83゜Example 121 Ac-Arg-Lys-I Ie-Leu-Phe-1, eu-Asp-Gl) ’ Manufacture of NHt The title compound was obtained in the same manner as above.

HPLC k' -3,81' (Harniyuton PRP-1' , 5% CH3CN/' 0.1% TFA -w@ 40% 7'H＞) 敞、S@　　　、　　　　　　　　　　sp　　1.00.　Gly　 1.05. Engineering 1e O,91,Leu 2.0BB Lys O, 94, krg 1.07. Phe 1.29° Example 122 Ac-Arg-Lys-11e-Leu-(4°NO,)Phe-Leu-A Production of sp-G1y-XHt A gyuhi compound was obtained in the same manner as described above.

HPLC: k’ = 3.87 (Hamilton PRP71, 5% CH3CN/O, 1% TrA 4,40 wires CM3CN/O, 1% TFA, 15 min,) Example 123 (4S,55)-4-(t-butyldemethylsiloxy)-5-(t-butoxyca Production of (S)-t-butoxycarboxylic acid)amino-6-funinylhexanoic acid Nylphenylalanine N-methoxy-N-methylamide a) The method of Org, Syn, 67.69 (1988) (=therefore the title compound was manufactured.

[α 365-'-, 34° (c=1. ethanol) (6S)-5- Oxo-6-(t-butoxycarbonyl-7-phenylhebut-1-ene b) Using Mg compound of 4-bromo-1-butene (X, implementation PI 1 2 3a) The title compound was obtained from the compound.

mpnia 4-75°C Co α Co D = -36,8' (c = 1. Ethanol) (5S) -4-oxo -5-(t-butoxycarbonyl)amino-6-phenylhexanoic acid C) The compound of Example 123 b) was stirred with benzene, water and acetic acid and the compound of Example 123 b) was stirred with tetrabutylene. Tylammonium and KMnOa were added to obtain the title compound.

[αko, = -36,1° (,5S)-4-oxo-5-(t-butoxycarbonyl)amino-6-phenylene Ruhexanoic acid benzyl ester d) Example 1 The compound of 123c was diazibicyclo Reaction with endecene and benzyl bromide gave the title compound.

mp: 35-87.5℃ This α co = 27.8” (4S, 5s)-4-(L-buterodimethyl 7)-5-(t-butoxy (cycarbonyl)amino-6-phenylhexanoic acid benzyl ester e) Reacting the compound of Example 123d) with t-butyldimethylsilyl chloride The title compound was obtained.

HPLC: R 9.18 min (451somer) 12.2 min (4 Risomer) (YMCA Yasun:” Boat x,, ++ , 1.0 ml/mln), (4R): (45)-3:l.

[a]o --14,4' (4S,5S)-4-(t-butyldimethylsiloquine)-5-(t-)ethoxy carbonyl)amino-6-phenylhexanoic acid f) Compound of Example L23e) was reduced by 5% palladium-carbon to obtain the title compound.

[α]. --24,7° Example 124 (5S)-benzyloxycarbonylalanyl-alanyl-(5-ami/-5- Production of phenyl-trans-hexene-3-en/yl)valyl-valinamide The title compound was obtained from the compound of Example 94 in the same manner as above.

Example 125 (5S)-benzyloxycarbonylalanyl-alanyl-(,5-ami/-6 -phenyl-cis-3,4-dihydroxy-hexanoyl)-valyl-valinea Manufacture of Mido The title compound was obtained from the compound of Example 125 in the same manner as above.

Example 126 (4S,5S)-alanylalanyl-(6-phenyl-5-amino-4-hydro (4S,5S)Boc- Aha-Ala-(6-phenyl-5-amino-4-t-butyldimethylsiloxy cyhexanoyl)-Va]-Va l-The title compound from Merryfield resin I got something.

Example 127 (4S,5S)-benzyloxycarbonylalanyl-alanyl-(6-phenylalanyl) (5-amino-4-hydroxyhexanoyl)valyl-valine The title compound was obtained from the compound of Example 126.

Example 128 (45,5S)-benzyloxycarbonylalanyl-alanyl-(6-phenylalanyl) -5-amino-4-hydroxyhexanoyl)-valyl-valyl-(5-amino-4-hydroxyhexanoyl) Production of mono-Boc-1,5-diaminopenkun a) 1.3 The title compound was obtained from -diaminopentane and di-tert-butyl dicarbonate.

Garlic MR (CDC Yu3): 61.43 (17H, br, s), 2.53 -2.83 (2H, m), 2.90-3-10 (2H-m) t 4 ．． 80-5.10 (l Hz by Nice) - MS (CI): m /z 237 (M+C1) di(4S, 5S)-benzyloxyca lbonylalanyl-alanyl-(6-phenyl-5-amino-4-hydroxyh) xanoyl) baryl-baryl- b) The compound of Example 128a) and the compound of Example 126 yielded the title compound. Ta.

Example 129 (4S,5S)-benzyloxycarbonylalanyl-alanyl-(6-phenylalanyl) -5-amino-4-hydroxyhexamethyl)-valyl-valyl-(5-gua Production of Nidinopentyl)amide The compound of Example 128 was mixed with ○-methylisourea. The title compound was obtained by reaction.

Example 130 (4S,5S)-alanyl-alanyl-(6-phenyl-5-amino-4-hydro Preparation of (4S , 5S)-Boc-Ala-Ala-(6-phenyl-5-amino-4-t-butyl Titled from (Tyldimethylsiloxyhexanoyl)-Va1-Va1-BHA The compound was obtained.

Example 131 (4S,5S)-alanyl-(6-phenyl-5-ami/-4-hydroquine Preparation of (4S,3S)Bo c-A　a-(5-7znyl-5-amino-4-t-butyldimethylsiloxy The title compound was obtained from (Sihexanoyl)-Va1-VaI-BHA.

Example 132 (4S,5S)-benzyloxycarbonylalanyl-alanyl-6-(phenylalanyl) -5(S)-amino-4(S)-hydroxyhexanoyl)-valyl-valine Preparation of amide The title compound was obtained from the compound of Example 130 in the same manner as described above.

Example 133 (4S,5S)-alanylalanyl-(6-7enyl-5-amino-4-hydro Production of (4S,5S)Boc-valine in the same manner as above. Aha-Ala-(6-phenyl-5-amino-4-t-butyldimethylsiloxy The title compound was obtained from Merryfield resin.

Example 134 (4S,5S)-benzyloxycarbonylalanylalanyl-(6-phenyl -Production of 5-amino-4-hydroxy(bovine shark)-valine The title compound was obtained from the compound of Example 133 in the same manner as above.

Example 135 (4S,5S)-benzyloxycarbonylalanylalanyl-6-(phenyl -5-amino-4-hydroxyhexanoyl)-valyl-(1-aminomethyl- Preparation of 2-methyl)propylamidecarbobenzyloxy-valinol a) The title compound was obtained from L-valinol and benzyl chloroformate.

1-phthaloylami7-2-carbobenzyloxy-3-methibutane b) Compound of Example 135a) The title compound was obtained.

1-t-Butoxycarbonylamino-2-carbobenzyloxydiamino-3-mer Chilbutane C) The title compound was obtained from the compound of Example 135b).

(4S,,5S)-benzyloxycarboalanyl-alanyl-(6-phenyl -5-amino-4-hydroxyhexanoyl)-valyl-(1-aminomethyl- 2-Methyl)propylamide d) The title compound was obtained from the compound of Example 135c). Ta.

(4S,5S)-benzyloxycarbonylalanyl-alanyl=(6-phenylalanyl) -5-amino-4-hydroxyhexanoyl)-valyl-(1-aminomethyl -2-methylbropyramidee) Compound of Example 135d) The title compound Obtained.

Example 136 (2R,5S)-2-(phenylmethyl)-5-t-butoxycarbonylamino -6-phenylhex-3-enoic acid production 2 (S) -t-futoquincarponi Ruamino-3-phenylpropertool a) Reacting Boc-L-phalaene and ethyl chloroformate and marking it Obtain a compound.

(2S)-1-(methylsulfonyl)oxy-2-t-butoxycarbonylamine Nor-3-phenylpropane b) Compound of Example 136a) and methanesulfonyl reaction with chloride gives the title compound.

(2S)-1-phenylthio 2-t-butoxycarbonylamino-3-phenylene Lupropane C) React thiophenol with sodium methoxide, and further react with Example 1. Reaction of compound 36b) gives the title compound.

(2S)-1-(Benzenesulfonyl)-2-t-butoxaldicarbonylamino 3-phenylpropane d) Reacting the compound of Example 136c) and m-chloroperbenzoic acid to form the title Obtain a compound.

(4R)-(2-oxo-4-(phenylmethyl)-3-oxazolidinyl)- 2-(phenylmethyl)-5-methylbent-3-enoate f) Following the method of Example 93C), prepare the title compound of Example 136e). obtain.

(2R)-2-(phenylmethyl)-5-methylbent-3-en-1-ol g) Treatment of the compound of Example 136 f) with LiAIH, gives the title compound.

(2R)-1-(tetrahydropyranyl)oxy-2-(phenylmethyl)-5 -Methyl-3-pentene h) Compound dihydropyran of Example 136(g) and Treatment with pyridinium p-)luenesulfonate gives the title compound.

(2S)-2-(phenylmethyl)-3-tetrahydropyranyloxypropio aldehyde i) Treatment of Example 136h) with ozone gives the title compound.

(2R,5S)-1-(tetrahydropyranyl)okine-2-(phenylmethyl ) 5-t-butoxycarbonylamino-6-phenylhex-3-ene j) The compound of Example 136 h) was treated with methyl-lithium, then the cytosulfonate was treated with sulfonate. dianion and treated with disodium hydrogen phosphate and sodium amalgam. The title compound is obtained.

(2R,5S)-2-(phenylmethyl)-5-t-butox dicarbonylamino -6-Phenylhex-3-enoic acidk) The compound of Example 136j) Treatment with reagents yields the title compound.

Example 137 (2R,5S)-alanylalanyl-2-(2-(phenylmethyl)-5-ami Nor-6-phenyl-trans-hexer-3-enoyl)-valyl-valinamide Manufacturing of The title compound is obtained from the resin-supported protected derivative prepared in the same manner as above.

Example 138 (2R,5S)-benzyloxycarbonylalanylalanyl-(2-(phenylalanyl) (methyl)-5-amino-6-phenyl-trans-hexer-3-enoyl) The compound of Example 137 for production of valyl-valinamide was (bonyloxy)succinimide to give the title compound.

Example 139 (6S)-6-(t-butofoxdicarbonyl)amino-7-phenyl-5-oxo -Production of hebutanoic acid Using the method of Example 12, but using 5-propylene instead of 4-bromo-1-butene. The title compound is obtained using mol-pentene.

Using the method of implementation IPI 19.34.76.77.78.88 and 135 , the following compound is obtained.

a) (5S,6S)-6-(carbobenzyloxy-alanylalanyl)ami7 -Two-phenyl-5-hydroxy-(1-oxo)butyl-valinme Tyl ester b) (53,6S)-6-(carbobenzyloxy-alanyl)a Mino-7-phenyl-5-hydroxy-(1-oxy)heptyl-valyl Bali methyl ester c) (5S,6S)-6-(carbobenzyl-oxalanylalanyl)amino -7-phenyl-5-hydroxy-(1-oxy)hebutyl-valyl Valinia Mido d) (5S,6S)-6-(carbobenzyloxy-alanyl)amino-7-phenyl phenyl-5-hydroxy-(1-oxy)hebutyl-valyl valineamide e) (5S,6S)-6-(carbobenzyloxy-alanylalanyl)amino -7-phenyl-5-hydroxy-(1-oxo)hebutyl-valyl Balino le f) (5S,6S)-6-(carbohensyloxy-"r=l)amino-7- Phenyl-5-hydroxy-(1-oxo)hebutyl-valyl valinol g) (5S,6S)-6-(carbobenzyloxy-alanylalanyl)amino -7-phenyl-5-hydroxy-(1-oxo)heptyl-valyl valine- ((S)l-amino-3-methylbutyl-2-)amide h) (53,6S)-6-(carbobenzyloxy-alanyl)amino-7-ph phenyl-5-hydrobutyl-valyl valine ((S) -1-amino-3-methylbutyl-2-)amide.

Example 140 By the method of Example 151, (2R,,5S)-2-methyl-5-(1-butoxy Preparation and implementation of cyclocarbonyl)amino-6-phenyl-4-oxo-hexanoic acid By the method of Examples 19.34.76.77.78.88 and 135, the following compounds get something

a) (2R,,4S,5S)-5-(carbobenzylo+cy7ranylalanyl )amino-6-phenyl-4-hydroxy-2-methyl-(1-oxy)hexy Rubalyl valine methyl ester b) (2R,4S,5S)-5-(carbobe alanyloxo-alanylalanyl)amino-6-phenyl-4-hydroxy-2 -Methyl-(1-oxo)hexyl-valyl valine 7mido c) (2R, 43, 5S)-5-(carbobenzyloxy-alanylalanyl)amino-6-pheny -4-hydroxy-2-methyl-(1-oxo)hexyl-valyl Balino (2R,4S,3S)-5(carbobenzyloxy-alanyl)amino- 6-phenyl-4-hydroxy-2-methyl-(1-oxo)hexyl-valyl Valinol f) (2R,4S,5S)-5-(carbobenzyloxy-arani lualanyl)amino-6-phenyl-4-hydroxy-2-methyl-(1-oxy So)hexyl-valyl valine-((S)-1-amino-3-methylbutyl-2 -) amide g) (2R,4S,5S)-5-(carbobenzyloxy-alanyl ) amido-6-phenyl-4-hydroxy-2-methyl=(1-oxo)hexy Rubaryl valine-((S)-1-amino-3-methylbutyl-2-)amide h) (2R,4S,5S)-5-(carpobenzyloxy-alanylalanyl) Amino-6-phenyl-4-hydroxy-2-methy/l/-(1-oxo)hex Syl-valyl valine(2R,4S,5S)-5-(carbobenzyloxy alanyl)amino-6-phenyl-4-hydroxy-2-methyl-(1-oxy ) Hexyl-baryl valine.

Example 141 (2R,5S)-2-benzyl-5-(t-butoxycarbonyl)amino-6- Production of phenyl-4-oxo-hexanic acid according to the method of Example 151, but, 3-bromo-2-methyl-1-benzyloxypropane instead of 3-bromo-2-methyl-1-benzyloxypropane Using 2-phenylmethyl-1-benzyloxypropane, the title compound is obtained.

Example 19.34.76.77.78.88 and 135 using the title compound According to the method, the following compounds are obtained.

a) (2R,4S,5S)-5-(carbobenzyloxy-alanylalanyl) Amino-6-phenyl-4-hydroxy-2-benzyl-(1-oxo)hexy Rubalyl valine methyl ester b) (2R,4S,5S)-5-(carbobe 5-alanyl)amino-6-phenyl-4-hydroxy-2-benzyl ndyl-(1-oxy)hexyl-valyl valine methyl ester c) (2R, 4S,5S)-5-(carbobenzyloxy-alanylalanyl)amino-6- Phenyl-4-hydroxy-2-benzyl-(1-oxo)hexyl-valyl Valinamide d) (2R,4S, 5S)-5-(carbobenzyloxia ranyl)amino-6-phenyl-4-hydroxy-2-benzyl-(1-oxo ) Hexyl-valyl Valinamide e) (2R,4S,5S)-5-(Carbohe amino-6-phenyl-4-hydroxy-2 -benzyl-(l-oxo)hexyl-valyl valinol f) (2R,4S, 5S)-5-(carbobenzyloxy-alanyl)amino-6-phenyl-4- Hydroxy-2-benzyl-(l-oxy)hexyl-valyl g) (2R,4S,5S)-5-(carbobenzyloxy-alanylalanyl)ami Nor-6-phenyl-4-hydroquine-2-benzyl-(1-oxo) Valyl valine-((S)-1-amino-3-methylbutyl-2-)amide h) (2R,4S,5S)-5-(carbobenzyloxy-alanyl)amino- 6-phenyl-4-hydroxy-2-benzyl-(1-oxy)hexyl-vari Valine-((S)-1-amino-3-methylbutyl-2-)amide f) (2R,4S,5S)-5-(carbobenzyloxy-alanylalanyl ) Amino-6-phenyl-4-hydroxy-2-benzyl-(1-oxo)hex Syl-valyl valine j) (2R,4S,5S)-5-(carbobenzyloxy -alanyl)amino-6-phenyl-4-hydroxy-2-benzyl-(1-o xo)hexyl-baryl valine.

Example 142 (3RS,5S)-3-inbutyl-5-(t-butoxycarbonyl)amino- Preparation of 6-phenyl-4-oxo-hexanoic acid to 4-hydroxy-6-methyl- Budol-l-ene a) Reacting isobutyraldehyde with allylmagnesium bromide The title compound is obtained.

4-Bromo-6-methylhept-1-eneb) The compound of Example 142a) Treatment with dichlorochloride and triethylamine gives the mesylate. LiB this Treatment with r gives the title compound.

(3R3,5S)-3-inbutyl-5-(t-butoxycarbonyl)amino- 6-phenyl-4-oxy-hexanoic acid C) Using the method of Example 12, but with: 4-bromo-6-metalhebutol-ene instead of 4-bromo-1-butene to obtain the title compound.

Following the method of Examples 19.34.76.77 and 78, the following compounds are obtained.

a1) (3SR,4S,5S)-5-(carbobenzyloxy-alanylara amino-6-phenyl-4-hydroxy-3-inbutyl-(1-oxy ) to beef sylubalyl valine methyl ester bl) (3SR, 4S, 5S)-5-(carbobenzyloxi-7-alanyl) Ami/-6-phenyl-4-hydroxy-3-inbutyl-(1-oxo)hex Syl-valyl valine methyl ester C1) (3SR,43,,5S)-5-( Carbohensyloxy-alanylalanyl)amino-6-phenyl-4-hydro xy-3-inbutyl-(1-oxo)hexyl-valyl valineamide dl) (3SR,4S,3S)-5-(carbobenzyloxy-alanyl)amino-6 -Phenyl-4-hydroxy-3-isobutyl-(1-1xo)hexyl-vari Valaldehyde.

Example 143 (3RS,5S)-3-methyl-5-(t-butoxycarbonyl)amino-6- Production of phenyl-4-oxo-hexanic acid Using the method of Example 142, however, Substituting acetaldehyde for inbutyraldehyde gives the title compound.

Using the method of Examples 19.34.76.77 and 78 the following compounds are obtained.

a) (3SR,4S,,5S)-5-(Carbohennrut+S7 Ranilalani ) amino-6-phenyl-4-hydroxy-3-methyl-(1-oxo)hexyl Syl-valyl valine methyl ester b) (3SR, 4S, 5S)-5-( Carbobenzyloxy-alanyl)amino-6-phenyl-4-hydroxy-3 -Methyl-(1-oxo)hexyl-valyl valine methyl ester c) (3S R,4S,5S)-5(carbobenzyloxy-alanylalanyl)amino-6 -Phenyl-4-hydroxy-3-methyl-(1-oxo)hexyl-valyl Valinamide d) (3SR, 4S, 5S)-5-(carbobenzyloxy- alanyl)amino-6-phenyl-4-hydroxy-3-methyl-(1-oxo ) Hexyl-valyl valinamide.

Example 144 (3RS, 5S)-3-benzyl-5-(t-butoxycarbonyl)amino Preparation of -6-phenyl-4-oxo-hexanoic acid Using the method of Example 142, Dashi, using phenylacetaldehyde instead of inbutyraldehyde, marking Obtain a compound.

Using the method of Examples 19.34.76.77 and 78, the following compounds are obtained: .

a) (3SR,4S,5S)-5-(carbobenzyloxy-alanylalanyl ) Amino7-6-phenyl-4-hydroxy-3-benzyl-(1-oxo)heki Syl-valyl valine methyl ester b) (3SR,4,S, 5S)-5- (carbobenyloxyalanyl)ami/-6-phenyl-4-hydroxy- 3-Benzyl-(1-oxo)hexyl-valyl valine methyl ester c) ( 3SR14S,,l5)-5-(carbobenzyloxy-alanylalanyl ndyl-(1-oxo)hexynrubalyl valinamide d) (3SR, 4S, 5S)-5-(carbobenzyloxy-alanyl)amino-6-phenyl-4 -Hydroxy-3-benzyl-(1-oxo) to bovine silubalyl valinamide .

Example 145 (,5S)-5-(t-butoxycarbonyl)amino-5-phenyl-4-o Production of xo-pentanoic acid Using the method of Example 12, but replacing Boc-phenylalanine with Boc - Using phenylglycine, the title compound is obtained.

Using the methods of Examples 34, 76, 77 and 78, the following compounds are obtained.

a) (53.4S) 5 (carbobenzyloxy-alanylalanyl)a Mino-5-phenyl-4-hydroxy-(1-oxo)pentyl-baryl Bali b) (5S,4S)-5-(carbobenzyloxy-alani) ) Amino-5-phenyl-4-hydroxy-(1-oxy)ventruva! J /l, valine methyl ester.

Example 146 (5S)-5-(t-butoxycarbonyl)amino-7-phenyl-4-oxo -Production of hebutanoic acid Follow the method of W Zeppetsu 12, add BoC- in place of Boc-phenylalanine. Using homophenylalanine, the title compound is obtained.

Using the methods of Examples 34, 76, 77 and 78, the following compounds are obtained.

a) (4S,5S)-5-(carbobenzyloxy-alanylalanyl)a Mino-7-phenyl-4-hydroxy-(1-oxo)hephthyluvalyl Bali methyl ester b) (4S,5S) -5- (carbobenzyloxy-ara Nyl)amino-7-7enyl-4-hydroxy-(1-oxo)heptyl-ba Lil valine methyl ester.

Example 147 (5S)-5-(carbobenzyloxyalanyl)amino-4-hydroxy-1 -Production of oxo-6-phenyl-beki/louvaline isobutyramide (5S)-5-(t-butoxy)ami/-1,4-dioxy-6-phenyl Silvaline Isobutyramide a) Compound of Example 12b) and NMM treatment with butyryl chloroformate. This is called L-valine isobutyramide. After reaction, the title compound is obtained.

(53)-5-(t-butoxycarbonyl)-4-hydroxy-1-oxo-6 - Phenyl-hexylvaline isobutyramide b) Compound of Example 147a) Reduction of the product with NaBH gives the title compound.

(5S)-5-Amino-4-hydro-1-oxo-6-phenyl-hexyl - Bawan Isobutyramide C) The title compound was obtained from the compound of Example 147b). Ru.

(,5S)-5-(carbobenzyloxyalanyl)amino-4-hydroxy- 1-oxo-6-phenylhexylvaline isobutyramide cl) Treat Cbz-alanine with inbutyryl chloroformate and perform Reaction of the compound of Example 147C) gives the title compound.

Example 148 (5S)-5-(carbobenzyloxyalanylalanyl)amino-4-hydro xy-1-oxo-6-phenylhexane valine isobutyramide In the same manner as Example 147d), cbZ-alanyl was used instead of Cbz-alanine. The title compound is obtained using lanin.

Example 149 to 5-(myristyryl)amino-4-hydroxy-1-oxo-6-phenyl- xyl valine isobutyramide Example 147c) Treatment with acid gives the title compound.

Example 150 5-(stearyl)amino-4-hydroxy-1-oxo-6-phenyl-hex Production of sylvaline isobutyramide Anhydrous stearic acid in place of myristic anhydride The title compound is obtained in the same manner as above using phosphoric acid.

Example 151 5-(carbobenzyloxyalanyl)amine-4-hydroxy-2-methyl- Production of 1-oxo-6-phenylhexylvalyl valine methyl ester 5-(t-butoxycarbonyl)amino-4-oxo-2-methyl-6-phenylene -1-benzyloxyhexane a) Following the method of Example 12a), but 3-bromo-2-methyl-1-benzyloxy instead of 4-bromo-1-butene using sea-propane (J. Med. Chem., 30.374, (1987)). , to obtain the title compound.

5-(t-butoxycarbonyl)amino-4-okine-2-methyl-1-hydro Roxy-6-phenylhexane b) Hydrogenolysis of the compound of Example 151a) The title compound is obtained.

5-(t-butoxycarbonyl)amino-4-oxo-2-methyl-6-phenylene Ruhexanoic acid C) The compound of Example 151b) (n-Bu), NBr and KMnOa Treatment with gives the title compound.

5-(carbobenzyloxyalanyl)amino-4-hydroboxy-oxo- 6-Funinyru to bovine sylbaryl valine methyl ester d) Following the method of Example 35a), the compound of Example 151c) was added to valylvaline. Coupling with a methyl ester and reducing the resulting ketone with NaBH, The amino group was deprotected using the method of Example 35c) and the Cbz-ara Coupling with nin gives the title compound.

Example 152 7-Methyl-5-(carbobenzyloxoalanylalanyl)amino-3,4- Dihydroxy-2-phenylmethyl-1-oxo-octyl valine isobutylene Ruamide 7-Methyl-4,5-○-(t-butoxycarbonyl)-N-impropylidene -3-Hydroxy-2-phenylmethyl-octanoic acid benzyl ester a”) 2,2-dimethyl-3-(t-butoxycarbonyl)-4-isopropyl -5-formyl-1,3-oxazolidine (J, Med, Chem, 30, 976 (1987)) with lithio-benzyl-3-phenylpropionate. The title compound is obtained.

7-methyl-5-(t-butoxycarbonyl)ami/-3,4-dioquine isopr Lopylidene-2-phenylmethyl-octanoic acid benzyl ester b) Treatment of the compound of Example 152a) with camphor-sulfonic acid to give the title compound get.

7-Methyl-5-(t-butoxycarbonyl)amino-3,4-cyoxyisobutylene Robylidene-2-funinylmethyl-octanoic acid C) Compound of Example 152b) Hydrogenation using palladium catalyst gives the title compound.

7-Methyl-5-(t-butoxyamino)-3,4-dioxy-impropylide -2-phenylmethyl-1-oxo-octylvaline isobutyramide d) Following the method of Example 87a) and using the compound of Example 152c), the title compound get something

7-Methyl-5-amino-3,4-dihydroxy-2-phenylmethyl-1-o Xo-octyl valine isobutyramide hydrochloride e) Same as Example 147c) The title compound is obtained from the compound of Example 152d).

7-Methyl-5-(carbobenzyloxyalanyl)amino-3°4-dihydro xy-2-phenylmethyl-1-oxo-octylvaline isobutyramide f) Analogously to the method of Example 147d), from the compound of Example 152e) the title Obtain the compound.

Example 153 (4S,5S)-5-(alanylalanyl)amino-4-hydroxy-1-oxy So-6-(4-hydroxyphenyl)hexyl-valyl valinol (4S,5S)-5-(carbobenzyloxo-alanylalanyl)ami/-4 -Hydroxy-1-oxo-6-(4-benzyloxyphenyl) to beef silva Lyruvaline a) Treatment of the compound of Example 72a) with KOH gives the title compound .

(4S,5S)-5-(carbobenzyloxy-alanylalanyl)amino-4 -Hydroxy-1-oxo-6-(4-benzyloxyphenyl)-hexyl- Valyl Valinol b) The compound of Example 153a) was converted into borane methyl sulfide. Treatment with gives the title compound.

(4S, 5S) 5 (alanylalanyl)amino-4-hydrobix 1-oxo-6-(4-hydroxyphenyl)-hexyl-valyl valinol C) Hydrogenation of the compound of Example 153'b) with a palladium catalyst to give the title compound. obtain.

Example 154 4-Methyl-3-[5-(alanylalanyl)amino-4-hydroxy-1-o xo-6-(4-hydroxyphenyl)-to-bovine sylvalylcoaminopentane- Preparation of 2-one 4-methyl-3-[5-(carbobenzyloxy-alanylara) amino-4-hydroxy-1-oxo-6-(4-benzyloxyphenylene) l) to bovine silubalyl]amino/-pentan-2-onea) Example 153a) Treatment of the compound with oxalyl chloride followed by cyazmethane gave the title compound. get.

4-Methyl-3-[5-(alanylalanyl)amino-4-hydroxy-1-o xo-6-(4-hydroxyphenyl)hequin-barylniami/benkun-2 −on b) Hydrogenation of the compound of Example 154a) over palladium catalyst to give the title compound. Ru.

Example 155 Ac-Arg-Ala-Ser-Gln-Asn-Tyr-Pro-Va I Production of -Val-NH, Boc-Tyr ( The title compound is obtained by replacing Boc-Phe with BrZ).

Example 156 Boc-His (Tos) in place of Boc-Ala using the method of Example 58 , Boc-~’al, Boc-11e, Boc-Lys(C1z), Boc -5e r (Bz I), Boc-D-Arg (TOS) or Boc-X The following compound is obtained using 1et.

His-5er-Gln-Tyr-E’ro-Val-Van-NF2 ;Val-5er-Gin-Asn-Tyr-Pro-Val-Val-NF2 　;Work1e-5e;-Gyun-Asn-τy=-p=r:+-v6you Vaney N! 42; Lys-5ay-Gin-λsn-τy knee? :o-VB1-Va -NH2;, D-) rg-5ex-GLn-Asn- Tyx-Pro-Val-Va knee NH2;5er-5er-Gin-Asn- Tyr-Pro-vax-Val-NF2; Met-5er-Gln-Asn- Tyr-Pro-Va Knee Va Knee NX2 Example 157 Following the procedure of Example 39, the following compound is obtained.

Ac-5e knee G1n-person sr+-Tyr-E’ro-Va knee Ala-NF2 ;Inc-5er-Gin-Asn-Tyr-Pro-Val-Gly-NF2; Input c-5er-Gin-Asn-Tyr-Pro-Val-Engineering1e-NH2; Input c-5ew-Gin-Asn-Tyr-Pro-Val-Leu-NF2; Implementation Example 158 Following the procedure of Example 39, the following compound was obtained: Ac-5ar-Gln-Ash -Tyr-Pro-Person La-Val-NH2; Ac-5er-Gin-Person s*- Tyr-Prci-Gly-Val-NF2; Enter c-5er-Gin-Asn- Tyr-Pro-11e-Val-NF2; entered c-5ar-Gin-person 5n-T yr-Hero-human rg-Val-NH2゜Ac-5er-Gln-Asn-T yr-Pro-Leu=Val-NF2　;Ac-5er-Gin-Asn-τ yr-Pro-Met-Val-N) 12 loquat example 159 According to the operations of implementation PJ40, obtain the following f and compound 2AC-5er-λ u a- H1S-Tyr-? ro-Va1-Vs=l-NH2; λe-5e knee λ1a- Val-Tyr-Pro-Va knee Val-NF, 2; Ac-5e knee Ala- Leu-Tyr-:’ro-Va knee Val-NH2; entered c-5ar-Ala- Tbr-Tyr-F'ro-Val-Val-NF2; λc-5er-Ala- Gl,n-Tyr-Pro-Val-Val-NF2 Example 160 (5S,4R3)-5-(t-butoxycarbonyl-serylalanylalanyl) Amino-4-hydroxy-1-oxo-6-phninylhexyl-valyl Ha II manufacturing The compound of Example 34d) is treated with potassium carbonate to give the compound of Example 34d).

Example 161 (33,4R3)-5-[(t-butoxy/carbonyl-serylalanylalanyl )-Aminony 4-hydroquine-1-dimention 6-phenylhexylvalyl Manufacture of valine-diethylamide; N-methylmorpholine , then Isobutilc C! treated with roformate and reacted with diethylamine The title compound is obtained.

Similarly, using n-butylamine instead of diethylamine, (5S, 4RS )-5-C(t-butofoxdicarbonylserylalanylalanyl)aminoco-4 -Hydroquine-1-oxo-6-phenyl to bovine silubalyl valinebutyramide Dora get le.

Example 162 (5S, 4R3) -5-[(t-butobovine dicarbonylcerylalanylalani ) Amino-[4-hydroxy-1-odine-6-7 phenylhexyl valine- Inbutyramide (53,4R3)-5-E (t-butoxycarbonyl-seryl alanylalanyl)aminoC-4-hydrocyanyl-oxo-6-phenyl Xyl valine methyl ester a) Implementation% Using the method of J34, substitute valine for baryl valine methyl ester. Using methyl ester, the title compound is obtained.

(5S,4R3)-5-:(t-butoxycarbonyl-serylalanylalanyl ) Amino[-4-hydroxy-1-oxo-6-phenylhexyl valine b) Obtain the title compound from the compound of Example 162a) using the method of Example 160. Ru.

(5S,4R3)-5-[(t-butoxycarbonyl-serylalanylalanyl ) Amino[-4-hydroxy-1-oxo-6-phenylhexyl valineiso Butyramide C) From the compound of Example 162b) using the method of Example 1.61 The title compound is obtained.

Example 163 Liposomal dosage unit composition Phosphatidylcholine (1,4g) and phosphatidylglycerol (0,6 g) was dissolved in 20% methanol (300 ml) in chloroform solvent and evaporated to dryness. Let it harden. A solution of peptide (30 mg in 200 ml of phosphate buffered saline) Add to this dry phospholipid film and equilibrate for 1-2 hours at room temperature. generated lipo Stir the somomolecule d solution to mix it evenly. The resulting suspension was poured into a 0.2μ polycarbonate Extrude through a carbonate filter 5 times to ensure uniform size distribution. Essential The suspension is then dialyzed or ultracentrifuged to remove unencapsulated peptide.

Example 164 Liposomal dosage unit composition In the beaker, cholesterol (49 mg) and olein! (0゜358g ) at 65°C for 20-30 minutes. While maintaining this 1 degree, phospholipids ( 1 g) slowly to ensure complete wetting by the oleic acid. Argini (0.22 g in 3.37 g of water) was added little by little at 40°C, mixed well and Let's call it Larry. Continue mixing at 40'C. After one week of equilibration, the peptide (15 0mg) and mix well to form a gel. If necessary, adjust the pH to 7.0 with acetic acid. Ru.

While stirring, add phosphorylated buffered saline (pH 7.4) little by little to 1 ml. The concentration is 200 mg of liposome gel per sample. Liposomes form naturally.

This method produces 5 g of liposome suspension. vA Single Dose Unit Liposo 1 g of suspension.

Example 165 Parenteral dosage unit composition A formulation containing 25 mg of the peptide of the invention is produced as follows.

25mg of peptide in 15ml of distilled water! dissolve in This solution was mixed under sterile conditions for 25 m Filter into one multi-dose ampoule and lyophilize.

For intravenous or intramuscular injection, this powder can be mixed with a 5% de-bovine strose solution (D5W). ) Reconstitute with 20ml. The dosage is determined by the injection volume. This solution It is also suitable for administration, for example in a bottle or bag, as in an EV drip.

Example 166 Oral dosage unit composition 35 mg of peptide was mixed with 7.5 mg of lactose and 5 mg of magnesium stearate. Combine, grind, and pass the resulting powder through a screen into hard gelatin capsules. and manufacture capsules for oral administration.

Although the present invention has been described in detail above, these are only specific examples, and the present invention is limited to these. However, all such variations are within the scope of the present invention.

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Claims (61)

[Claims] 1. Formula (I): A-B-(Q)a-(C)b-(D)-M-(W)d-(X)e-Y-Z(I) [In the formula, A is BocNH, CbzNH, H, R′R′′N, R′′CONR′ or or DnsNH, or when a, b and c are O and Y is a covalent bond, A is H, Boc, Cbz, R″ or R″CO: B is one or more D or L-ami Glx, A sx, Ala, β-Ala, Arg, Glly, He, Leu, Lys, Se r, Thr, Val, Met or His; Q is a D or L-amino acid, Ser, Thr, Asp, His, Cys, Ar g or Ala; W is Pro or Δ3-dehydro-Pro; X is Ala, Gly, He, Le u, Val, Met, Lys, Glx or Asx; Y is one or more D or L-amino acids or a covalent bond; Z is CO2R'', CON R'R''''', COR', CH2OH, CH2NR'R''''' or H Alternatively, if d and e are 0 and Y is a covalent bond, Z is OR′′′′ or NR′R′ '''; a, b, c, d and e are each independently 0 or l, provided that c and e cannot be 0 at the same time; M is Cha, Phe (4'Ra) or -NHCHR1R2-; where R1 is Independently, C1-5Alk, (CH,)nSC1-5Alk, (CH2)nOC 1-5Alk, (CH2)nC3-7cycloalkyl or (CH2)mC6H 4-Ra; Ra is halogen, OR', NO2, NH2 or H; (CH2)n-, CHR3 (CH2)mCO-, CO(CH2)mCO-, CHR3CHR3, (CH2) mCO-,CHR3CHR3,CHR1(CH2)mCO-,CHR3CHR1 CHR'(CH2)mCO-, CHR3CHR'CHR1(CH2)mCO-, COCR1CHR'(CH2)mCO-, COCF2CR'R''(CH2)m CO-, COCF2(CH2)mCO-, CH=CR'CHR''(CH2)m CO-, COCH=CR'(CH2)mCO-, CH2S(O)pCHR'(C H2)mCO-, CH2NR'CHR''(CH2)mCO-, P=O(OR' )(CH2)mCO-,(CH2)n-,CHR3(CH2)mCO-,CO( CH2)mCO-,CHR3CHR3, (CH2)mCO-,CHR3CHR3 ,CHR1(CH2)mCO-,CHR3CHR1CHR'(CH2)mCO- ,CHR3CHR'CHR1(CH2)mCO-,COCR1CHR'(CH2 ) mCO-, COCF2CR'R''(CH2) mCO-, COCF2(CH2 ) mCO-, CH=CR'CHR''(CH2)mCO-, COCH=CR'( CH2)mCO-, CH2S(O)pCHR'(CH2)mCO-, CH2NR 'CHR''(CH2)mCO-, P=O(OR')(CH2)mCO-, ▲number There are formulas, chemical formulas, tables, etc. ▼, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼, ▲ Mathematical formulas, There are chemical formulas, tables, etc. ▼, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼, ▲ Mathematical formulas, chemistry There are formulas, tables, etc. ▼▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ or ▲ Mathematical formulas, chemical formulas , tables, etc. ▼; R3 and R3' are each independently OH, H or NH2: R4 is OH, H, NH2 or =O; m is independently 0, 1, 2 or 3; n is independent 1 or 2; p is 0, 1 or 2; R' is H or C1-5Alk; R'' is H or C1-16Alk; R'''' is C1-5Alk, C3-6 cycloalkyl, (CH2)nC6H5 , (CH2)nC5H4N, (CH2)n〇H, (CH2)nNH2 or (C H2)nNHC(NH)NH2 means. ] A peptide represented by or a pharmaceutically acceptable salt thereof. 2. B is Ala, Gly, Va1, Ile, Leu, Met, Met, His, From the group consisting of Lys, Arg, Glx, Asx, Cys, Ser and Thr Claim 1 which is one or two selected D or L-amino acids or a covalent bond. Peptides described. 3. Y is Ala, Gly, He, Leu, Met, Va1, Arg, Lys, 1-3 selected from the group consisting of Thr, Ser, Cys, Glx and Asx The peptide according to claim 1, which is a D or L-amino acid or a covalent bond. 4. B is Ala, β-Ala, Gly, Va1, He, Leu, Met, Hi a group consisting of s, Lys, Arg, Glx, Asx, Cys, Ser or Thr one or two D or L-amino acids selected from or a covalent bond, where Q is S D or L-amino selected from the group consisting of er, Thr, Asp and His The peptide according to claim 3, which is an acid. 5. D is Glx, Asx, Ser, Ala, β-Ala, Ile, Leu, Va 5. The peptide according to claim 4, selected from the group consisting of 1 and Met. 6. Y is from Ala, Gly, Ile, Met, Asx, Arg and Val. The peptide according to claim 5, which is an amino acid selected from the group consisting of: or a covalent bond. 7. The peptide according to claim 6, wherein M is -NHCHR1R2-. 8. 8. The peptide according to claim 7, wherein d is 0. 9. R1 is CH2C6H4-Ra, CH2C6H11 or C1-5Alk The peptide according to claim 8. 10. R2 is CHR3CHR3'(CH2)mCO-, CHR3CHR3'CHR1CO-, CHR3CHR1CHR'CO-,CHR3(CH2)mCO-,CO(CH2 )mCO-, COCR1CHR'(CH2)mCO-,CHR3CF2(CH2 )mCO-, -COCF2(CH2)mCO-,P=O(OR')(CH2)m CO-, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼, ▲ There are mathematical formulas, chemical formulas, tables, etc. 10. The peptide according to claim 9, which is ▼ or ▲ has a mathematical formula, chemical formula, table, etc. 11. 11. The peptide according to claim 10, wherein B is a covalent bond. 12. A claim in which C and D are selected from the group consisting of Gln, Asn and Ala. Peptide according to 11. 13. Q is a D or L-amino acid selected from the group consisting of Ser and Thr; 13. A peptide according to claim 12. 14. 14. The peptide according to claim 13, wherein X and Y are Val. 15. 15. The peptide according to claim 14, wherein C is Gln and D is Asn. 16. 15. The peptide according to claim 14, wherein C and D are Ala. 17. The peptide according to claim 5, wherein M is (4'Ra)Phe. 18. 18. The peptide according to claim 17, wherein at least one of X and Y is Val. 19. Acetyl-seryl glutaminyl asparaginyl tyrosyl prolyl valilva Linamide; Acetyl-serylalanylalanyltyrosylprolylvalyl valineamide; also teeth Acetyl-arginylalanylserylglutaminyl asparaginyl tyrosylpro 19. The peptide according to claim 18, which is lylvalylvalinamide. 20. The peptide according to claim 11, wherein R1 is -CH2C6H5Ra. 21.5-(carbobenzyloxy-alanylalanyl)amino-6-phenyl -4-hydroxy-(1-oxo)hexylvalylvalinol; Carbobenzyloxy-alanyl-(3-hydroxy-5-amino-6-phenylene) ) hexanoyl-valyl-(4-aminomethyl)pyridine; 5-(carbobenzyloxyalanylalanyl)amino-4-hydroxy-6- Phenyl(1-oxo)hexylubaline isobutylamide; 2-(acetyl-serylglutaminyl asparaginyl)amino-3-phenylp ropyruprolylvalylvalinamide; 2-(serylglutaminyl asparagini) 4-(acetyl)amino-3-phenylpropylprolyl valyl valineamide; (ru-cerylglutaminyl asparaginyl)amino-3-hydroxy-1-oxo -5-phenylpentylprolyl valyl valinamide; 4-(acetyl-serylglutaminyl asparaginyl)amino-3-hydroxy -1-oxo-5-phenylpentylvalylvalinamide; or 4-(serylglutaminylasparaginyl)amino-3-(S)-hydroxy- 1-oxo-5-phenylpentylvalyl valinamide; and a pharmaceutically acceptable salt thereof. 22.2-((2-(t-ptoxycarbonyl-serylalanylalanyl)ami (-1-oxo-3-phenylpropyl)cyclopentylcarbonylvalylmethylene ester: 2-(2-(serylalanylalanyl)amino-1-oxo-3- phenylpropyl)cyclopentylcarbonylvalylvaline methyl ester; 2-(2-(t-ptoxycarbonyl-serylalanylalanyl)amino-1- Hydroxy-3-phenylpropyl) cyclopentylcarbonyl valyl valinme Tyl ester; 2-(1-hydroxy-3-phenyl-2-(serylalanyl) ranyl) aminopropyl) cyclopentyl carbonyl valylvaline methyl ester Le: 2-((1-methoxy-1-(2-phenyl-1-(t-ptoxycarbonyl- serylalanylalanyl)amino)ethyl)phosphinyl)cyclopentylcal Bonyl-valylvaline methyl ester 2-((1-hydroxy-1-(2-phenylene) Nyl-1-(serylalanylalanyl)amino)ethyl)phosphinyl)cyclo Pentylcarbonyl-valylvaline methyl ester; 5-(t-ptoxycarboxylate) amino-4-hydroxy-1-oxo-6-ph Phenylhexylvalylvaline methyl ester: 5-(t-ptoxycarbonyl-serylalanylalanyl)amino-4-hydro xy-1-oxo-6-(4-hydroxyphenyl)hexylvalylvalinme Tyl ester; 4-(t-ptoxycarbonyl-serylalanylalanyl)ami Nor-2,2-difluoro-3-hydroxy-1-oxo-5-phenylpentyl -valylvaline methyl ester; 4-(serylalanylalanyl)amino-2, 2-difluoro-3-hydroxy-1-oxo-5-phenylpentyl-valyl Valine methyl ester; 4-(serylalanylalanyl)amino-2,2-difluoro-1,3-dioki So-5-phenylpentyl-valylvaline methyl ester; 5-(t-ptoxycarbonylserylalanylalanyl)amino-4-hydroxy C-6-(4-hydroxy)phenyl-1-oxohexylvalylamide 5-((carbobenzyloxy-D-seryl)alanylalanyl)amino-4- Hydroxy-1-oxo-6-phenylhexylvalylvaline methyl ester ; 5-((D-seryl)alanylalanyl)amino-4-hydroxy-1-oxo -6-phenylhexylvalylvaline methyl ester; 5-(t-ptoxycarbonylserylalanylalanyl)amino-6-(4-hyperoxycarbonyl) droxy)phenyl-4-hydroxy-1-oxohexylvalylvaline; 5-(serylalanylalanyl)amino-6-(4-hydroxy)phenyl-4 -hydroxy-1-oxo-hexylvalylvaline; or 2-((1-hydroxy-2-(serylalanylalanyl)amino-3-cyclo Hexyl)propylcyclopenkune carbonyl-valylvaline methyl ester or a carboxamide thereof; and a pharmaceutically acceptable salt thereof. Peptides described. 23. (1R,2R)-2-((1S,2S)-(1-hydroxy-3-phenylene) -2-(serylalanylalanyl)aminopropyl)cyclopentylcarbonyl Claim 22 which is rubaryruvaline methyl ester and a pharmaceutically acceptable salt thereof. Peptides described. 24. 12. The peptide according to claim 11, wherein a is 0. 25. 25. The peptide according to claim 24, wherein R2 is -CHOCH2CH2CO-. 26. 25. The peptide according to claim 24, wherein b is 0. 27. 25. The peptide according to claim 24, wherein Y is a covalent bond. 28. 26. The peptide according to claim 25, wherein D is Ala and X is Val. 29. 27. The peptide according to claim 26, wherein c is 0. 30. (4RS,5S)-5-((t-ptoxycarbonyl)isoleucyl)a Mino-7-methyl-4-hydroxy-1-oxo-octyl-leucyl-aspa Laginate methyl ester; (4RS,5S)-5-((t-ptoxycarbonyl ) isoleucyl) amino-7-methyl-4-hydroxy-1-oxo-octyl -Leucyl-(0-benzyl)aspartic acid methyl ester: (4S,5S) -5-(carbobenzyloxyalanyl)amino-4-hydroxy-1-oxo -6-phenylhexylvalylvaline methyl ester: (4S,5S)-5-(alanyl)amino-4-hydroxy-1-oxo-6- Phenylhexylvalylvaline methyl ester; (4S,5S)-5-(t-ptoxycarbonylalanyl)amino-4-hydro xy-1-oxo-6-(4-hydroxyphenyl)hexylvalylvalinme Tyl ester: or (4S,5S)-5-alanylamino-4-hydroxy- 1-oxo6-(4-hydroxyphenyl)-hexylvalylvaline methyl ester Stell; and a pharmaceutically acceptable salt thereof. 31. 28. The peptide according to claim 27, wherein e is 0. 32. (4S,5S)-5-(methoxycarbonyl-alanylalanyl)amino -6-phenyl-4-hydroxy-1-oxo-hexylvalylvaline; (4S,5S)-5-(carbobenzyloxyalanylalanyl)amino-6- Phenyl-4-hydroxy-1-oxo-hexylvalylvaline (4S,5S)-5-(alanylalanyl)amino-6-phenyl-4-hydro xy-1-oxo-hexylvalylvaline; (4S,5S)-5-(alanyl alanyl)amino-4-hydroxy-1-oxo-6-(4-hydroxyphenylene ) hexylvalylvaline methyl ester; (4S,5S)-5-[(carbobenzyloxy-β-alanyl)alanyl]a Mino-4-hydroxy-1-oxo-6-(4-hydroxyphenyl)hexyl -valylvaline methyl ester; (4S,5S)-5-[(β-alanyl)ara Amino-4-hydroxy-1-oxo-6-(4-hydroxyphenyl) Hexylvalylvaline methyl ester; (4S,5S)-5-(alanylalanyl)amino-4-hydroxy-1-ox So-6-cyclohexylhexylvalylvaline methyl ester: (4S,5S)-5-(carbobenzyloxyalanylalanyl)amino-4- Hydroxy-1-oxo-6-fuconylhexylvalylvaline methyl ester ;or (4S,5S)-5-(alanylalanyl)amino-4-hydroxy-1-ox So-6-phenylhexylvalylvaline methyl ester or its carboxylic acid The peptide according to claim 28, which is cyamide; and a pharmaceutically acceptable salt thereof. 33. (4RS,5S)-5-amino-6-(4-hydroxy)phenyl-4- Hydroxy-1-oxo-hexylvalylvaline benzyl ester: (1R,2R)-2-(((1S,2S)-1-hydroxy-2-amino-3- cyclohexyl)propyl)cyclopentanecarbonyl-valylvaline methyl ester Stell: (4S,5S)-5-amino-4-hydroxy-1-oxo-6-phenylhex sylvalylvaline methyl ester; or (4S,5S)-5-(t-ptoki (cycarbonyl)amino-4-hydroxy-1-oxo-6-phenylhexyl Valylvaline methyl ester or its carboxamide; and its pharmaceutical 30. The peptide of claim 29, which is an acceptable salt. 34. (4S,5S)-5-(alanyl)amino-4-hydroxy-1-oxo -6-(4-hydroxyphenyl)hexylvalylvaline methyl ester and 31. The peptide according to claim 30, which is a pharmaceutically acceptable salt thereof. 35. (4S,5S)-5-(carbobenzyloxyalanyl)amino-4-hydroxy Droxy-1-oxo-6-phenylhexylvalylvaline methyl ester 31. The peptide according to claim 30. 36. (4S,5S)-5-(carbobenzyloxyalanylalanyl)amino -4-Hydroxy-1-oxo-6-phenylhexylvalylvaline methyl ether 33. The peptide according to claim 32, which is a stellate. 37. (4S,5S)-5-(alanylalanyl)amino-4-hydroxy-1 -Oxo-6-phenylhexylvalylvaline methyl ester and its pharmaceuticals 33. The peptide of claim 32, which is an acceptable salt. 38. A pharmaceutical composition comprising the peptide according to claim 9 and a pharmaceutically acceptable carrier. . 39. A pharmaceutical composition comprising the peptide according to claim 9 and a ribosome carrier. 40. The peptide according to claim 9, azidothymidine and a pharmaceutically acceptable carrier. A pharmaceutical composition comprising: 41. A pharmaceutical composition comprising the peptide according to claim 20 and a pharmaceutically acceptable carrier. thing. 42. A pharmaceutical composition comprising the peptide according to claim 22 and a pharmaceutically acceptable carrier. thing. 43. A pharmaceutical composition comprising the peptide according to claim 28 and a pharmaceutically acceptable carrier. thing. 44. A pharmaceutical composition comprising the peptide according to claim 32 and a pharmaceutically acceptable carrier. thing. 44. in a mammal comprising administering an effective amount of the peptide according to claim 9. How to control viral diseases. 45. in a mammal comprising administering an effective amount of the peptide according to claim 9. How to control retroviral diseases caused by cancer. 46. administering an effective amount of the peptide according to claim 9 and azidothymidine; A method for suppressing retroviral diseases in mammals. 47. A method for treating HIV infection comprising administering an effective amount of the peptide according to claim 9. How to treat caries. 48. administering an effective amount of the peptide according to claim 9 and azidothymidine; A method for treating medical conditions associated with HIV infection. 49. The peptide according to claim 17 is infused with a retrovirus protease. Retroviral profiling consists of Method for assaying tease activity. 50. The peptide according to claim 19 is infused with a retroviral protease. Retroviral profiling consists of Method for assaying tease activity. 51. formula ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [wherein R1 is independently C1-5Alk, -(CH2)nSC1-5Alk, -(CH2)nOC1-5Alk, -(CH2)nC3-7cycloalkyl or -(CH2)mC6H4-Ra; Ra is halogen, NO2, OH, OC1-5 Alk, NH2 or H; R2 is (CH2)n-X, (CH2)nCO-X, C HR3CHR1CHR'(CH2)mCO-X, CHR3CF2(CH2)mC O-X, COCF2CR'R''(CH2)mCO-X, CHR3(CH2)m CO-X, CO(CH2)mCO-X, COCR1CHR'(CH2)mCO- X, ▲There are mathematical formulas, chemical formulas, tables, etc.▼,▲There are mathematical formulas, chemical formulas, tables, etc.▼, ▲There are mathematical formulas, chemical formulas, tables, etc.▼,▲There are mathematical formulas, chemical formulas, tables, etc.▼,▲Numbers There are formulas, chemical formulas, tables, etc. ▼, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ or ▲ Numbers There are formulas, chemical formulas, tables, etc. ▼; R3 is H, NH2 or OH; R4 is H, NH2, OH or =O; R5 is H; R6 is CH3CO, C1-5Alk, Dns, Cbz or Boc, or R 5 and R6 together represent phthalimide; X is H, halogen, OH, OC1- 5Alk, NHR'R'' or OCOC1-5Alk R' and R'' are H or C1-5Alk; m is independently 0, 1, 2 or 3 ; n is independently 1 or 2; However, R1 is phenylmethylene, isobutyl or cyclohexylmethylene In the case, R2 is CHR3CH2CO-X, COCH2CO-X, COCF2CO- If R1 is isobutyl and not X or CHR2CF2CO-X, then R2 is C H(OH)CH2CH2CO-X or COCH2CH2CO-X] Compounds shown. 52. formula ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [Wherein, R1 is C1-5Alk, -(CH2)nSC1-5Alk, -(CH2 )nOC1-5Alk, -(CH2)nC3-7cycloalkyl or -(CH 2) mCoH4-Ra: Ra is halogen, NO2, OH, OC1-5Alk, NH2 or H; m is 0, 1, 2 or 3; n is 1 or 2; R4 is H, NH2, OH or =O; R5 is H; R6 is CH3CO, C1-Alk, Dns, Cbz or Boc, or R5 and R6 together form phthalimide; X is H, halogen, OH, OC1-5Al k, NHR'R'' or OCOC1-5Alk; R' and R'' mean H or CI-5Alk] Claim 51 Compounds described. 53. formula; ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [Wherein, Ra is halogen, NO2, OH, OC1-5Alk, NH2 or H; R4 is H, NH2, OH or =O; R5 is H; R6 is CH3CO, C1-5Alk, Dns, Cbz or Boc or R5 and R6 together form phthalimide; X is H, halogen, OH, OC1-5 Alk, NHR2'R'' or OCOC1-5Alk; R' and R'' mean H or C1-5Alk] Claim 52 Compounds described. 54. formula: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [Wherein, R1 is C1-5Alk, -(CH2)nSC1-5Alk, -(CH2 )nOC1-5Alk, -(CH2)nC3-7cycloalkyl or -(CH 2) mC6H4-Ra; Ra is halogen, NO2, OH, OC1-5Alk, NH2 or Hm is 0, 1 , 2 or 3; n is 1 or 2; R5 is H; Ra is CH3CO, C1-5Alk, Dns, Cbz or Boc or R5 and Ra together form phthalimide; X is H, halogen, OH, OC1-5 Alk, NHR'R'' or OCOC1-5Alk; R' and R'' mean H or C1-5Alk] Claim 51 Compounds described. 55. formula: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [Wherein, Ra is halogen, NO2, OH, OC1-5AIk, NH2 or H; R5 is H; R6 is CH3CO, C1-5Alk, Dns, Cbz or Boc or R5 and R6 together form phthalimide; X is H, halogen, OH, OC1-5 Alk, NHR'R'' or OCOC1-5Alk; R' and R'' mean H or C1-5Alk] Claim 54 Compounds described. 56. formula: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [Wherein, R1 is C1-5Alk, -(CH2)nSC1-5Alk, -(CH2 )nOC1-5Alk, -(CH2)nC3-7cycloalkyl or -(CH 2) mC6H4-Ra: R1' is C1-5Alk, -(CH2)nSC1-5Alk, -(CH2)nO C1-5Alk, -(CH2)nC3-7cycloalkyl or -(CH2)m C6H4-Ra; m is 0, 1, 2 or 3: n is 1 or 2; Ra is halogen, NO2, OH, OC1-5Alk, NH2 or HR4 is OH , NH2, H or =O; R5 is H; R6 is CH3CO, C1-5Alk, Dns, Cbz or Boc or R5 and R6 together form phthalimide; X is H, halogen, OH, OC1-5 Alk, NHR'R'' or OCOC1-5Alk; R' and R'' mean H or C1-5Alk. However, R1 is isobutyl It is a group other than chill] 52. The compound according to claim 51. 57. formula: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [Wherein, Ra is halogen, NO2, OH, OC1-5Alk, NH2 or H; R4 is OH, NH2, H or =O; R5 is H: R6 is CH3CO, C1-5Alk, Dns, Cbz or Boc or R5 and R6 together form phthalimide; X is H, halogen, OH, OC1-5 AIk, NHR'R'' or OCOC1-5Alk; R' and R'' mean H or C1-5Alk] Claim 56 Compounds described. 58. 58. The compound according to claim 57, wherein R4 is OH or =O, and Ra is H or OH. thing. 59. formula: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [Wherein, R1 is C1-5Alk, -(CH2)nSC1-5A1k, -(CH2 )nOC1-5Alk, -(CH2)nC3-7cycloalkyl or -(CH 2) mC6H4-Ra: m is 0, 1, 2 or 3; n is 1 or 2; Ra is halogen, N02, OH, OC1-5Alk, NH2 or H; R5 is H ; R6 is CH3CO, C1-5Alk, Dns, Cbz or Boc or R5 and R6 together form phthalimide; X is H, halogen, OH, OC1-5 Alk, NHR'R'' or OCOC1-5Alk; R' and R'' mean H or C1-5Alk] Claim 51 Compounds described. 60. formula: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [Wherein, Ra is halogen, NO2, OH, OC1-5Alk, NH2 or H; R5 is H; R6 is CH3CO, C1-5Alk, Dns, Cbz or Boc or R5 and R6 together form phthalimide; X is H, halogen, OH, OC1-5 Alk, NHR'R'' or OCOC1-5Alk; R' and R'' mean H or C1-5Alk] Claim 59 Compounds described. 61. Peptide instead of one or two amino acids that are subject to proteolysis by proteases. 52. Said protease, characterized in that it incorporates the compound according to claim 51 in said protease. A method for producing protease inhibitory activity in the peptide, which is a substrate for.