JPH041140A - Method for producing 3-amino-2-oxo fatty acid derivative - Google Patents

Abstract

PURPOSE:To produce a physiologically active substance having 3-amino-2- oxocarbonyl group by oxidizing a 3-amino-2-hydroxyfatty acid derivative with an oxidizing agent such as DMSO and, as necessary, eliminating protecting group. CONSTITUTION:The objective compound having a partial structural formula II can be produced by oxidizing a compound having partial structural formula I (R1 is saturated or unsaturated hydrocarbon group) and, as necessary, eliminating protecting group. The reaction is preferably carried out under a mild condition not to exert adverse effect to the compound except for the oxidation at 2-site. Preferably, DMSO is used as the oxidizing agent and a combination of a weak base and an acid (preferably salt of trifluoroacetic acid and pyridines) as a catalyst. Further, it is preferable to use a carbodiimide, acetic anhydride, oxalyl chloride, etc., in combination with the above compounds, The reaction is carried out at 0-40 deg.C for 3-50hr in DMSO or other solvent.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field 1 The present invention relates to endopeptidases (particularly serine enzymes or
The present invention relates to a method for producing a novel synthetic peptide that has inhibitory activity against enzymes classified as HIW molecules.

[Prior art] Peptide aldehydes such as leupeptin and elastatinal have been known as natural products of low molecular weight peptides that inhibit endopeptidase, and many peptide aldehydes have been developed based on these. Synthesized as an inhibitor.

Thompson has shown that these peptide aldehydes exhibit inhibitory activity by binding to N# and occupying the reaction site between the aldehyde and the peptide carbonyl group on the enzyme.
It has been pointed out by R.C. (Biochemist
ry, Vol. 12, pp. 47-51 (1973)).

[Problem to be solved by the invention Since the carbonyl group, which is the active center of the cobebutide aldehyde group, is located at the C-terminus of the peptide chain, the molecular structure of the cobebutide aldehyde group can be changed to selectively bind to enzymes. Only the N-terminal side of the active center carbonyl group. Poststatin (compound No. 01 described below), which was previously invented by the present inventors, has a 3-amino-2-oxocarbonyl group with the following partial structural formula % formula % in the peptide chain, and has amino acid residues on both sides of the 3-amino-2-oxocarbonyl group. It has a structure. If the amino acid residues on both sides can be changed appropriately and combined with the structural part containing this carbonyl group, it can be synthesized.
It is believed that it is possible to produce substances that selectively exhibit inhibitory activity against various peptidases.

An object of the present invention is to provide a new method for producing a physiologically active substance containing the above partial structure.

In the past, peptides containing 3-amino-2-keto acids were
Examples have been reported. Hori et al. reacted C-
It has been reported that elastase can be inhibited by synthesizing a peptide having ethyl 3-amino-2-oxo-pentanoate or benzyl ester at the end.
s, 5structure and Funct+on,
Proceedings of the N1n
thAleriCan pet+tide Sym
posiugi, DD 8 1 9 -8 22
). However, because they used the Dakin-West reaction, the root of the 3-amino group was racemized, and this method had problems such as being able to synthesize only C-terminal esters.

[Means for Solving the Problems] Therefore, the present inventors have made extensive studies to overcome the above problems, and as a result, as a precursor of a physiologically active substance having the above partial structure, the following partial structural formula (in the formula R1 represents a saturated or unsaturated hydrocarbon group) The functional groups other than the human Ooxy group at the 2-position of the 3-amino-2-hydroxy fatty acid derivative are protected as necessary. ), then
This 3-amino-2-hydroxy fatty acid derivative is oxidized to obtain 3-=amino-2-oxofat I having the following partial structural formula (wherein R1 is the same as above). By obtaining the derivative and removing the protective groups of the functional groups as necessary,
The present invention was completed by discovering that the physiologically active substance '1K (or an intermediate thereof) having the above-mentioned 3-amino-2-oxocarbonyl group can be obtained.

The oxidation reaction of the present invention is preferably carried out under mild conditions so as not to have any adverse effects other than the oxidation at the 2-position of the above partial structural formula.

For example, DMSO (dimethyl sulfoxide) as an oxidizing agent
Oxidation using is preferred.

When using DMSO, the catalyst is usually a weak base (for example, pyridines with a PKa' of about 7.5 or less) and an acid (
For example, using a combination catalyst with a PKa of about 3 or less, such as trifluoroacetic acid, phosphoric acid, etc., preferably a salt of trifluoroacetic acid and pyridines, It is preferable to use propylcarbodiimide), acetic anhydride or oxalyl chloride in combination.

The reaction may be carried out using DMSO as a solvent, or may be carried out in another inert solvent as long as it does not adversely affect the reaction. It is preferable that the reaction temperature is approximately 0°C to 40°C. The reaction time varies depending on the conditions, but is usually 3 to 50 minutes.
The time is about 1 to 1 minute.

The physiologically active substance (or its intermediate) obtained by the method of the present invention has the partial structural formula % formula % [111 (wherein R1 is the Same as , Y represents a residue obtained by removing hydrogen from the amino group of an amino acid or peptide or their ester.When Y has a functional group, it may be protected with a protecting group. Preferably general formula X-N
H-CII-CO-CO-Y [
IV ] R1 <In the formula, R and Y are the same as above, X represents a residue obtained by removing OH from an amino acid or a carboxyl group of a peptide, and when X has a functional group, it may be protected. ) Poststatin and its related compounds can be mentioned.

These poststatins and their analogues are represented by the general formula (in the formula, X' and Y' represent X and Y with protected functional groups, respectively, X and Y are the same as above, and R1 is the same as above). It can be obtained by oxidizing a 3-amino-2-human O-oxy fatty acid derivative.

The substance represented by the general formula [Vl can be produced by a conventional peptide synthesis method using 3-protected-amino-2-human 0xy-propionic acid having a substituent corresponding to R1 as an amino acid.

If a general liquid phase method is used, for example when catalytic hydrolysis is used for the final deprotection, amino acid benzyl ester or benzyl ester of a peptide, e.g. p-methoxybenzyloxycarbonyl-3-amino-2-hydroxy- Fatty acids or t-butoxycarbonyl-Oxy-fatty acids, etc., are mixed in a solvent commonly used for peptide synthesis, such as dimethylformamide, and a condensing agent, such as dicyclohexylcarbodiimide, carbonyl carbide, posiimide, N-(3- Using carbodiimides such as dimethylaminopropyl)-N'-ethylcarbodiimide or a salt thereof, or a combination thereof with a condensation aid, such as 1-hydroxybenzotriazole (HoBt), N-hydroxy'succinimide, etc. The p-methoxybenzyloxycarbonylbonyl (BOC) group was removed using a selective deprotecting agent such as trifluoroacetic acid, and the general formula [Vl
Depending on Y' and X' in , it can be synthesized by condensation using a conventional method.

Also, if trifluoroacetic acid is used for final deprotection,
Amino Ilt-butyl ester is condensed with p-methoxybenzyloxycarbonyl-hydroxy-fatty acid or benzyloxycarbonyl-like, p-methoxybenzyloxycarbonyl group or benzyloxycarbonyl group is removed by catalytic hydrogenolysis, and then the general formula (Il) is condensed with amino Ilt-butyl ester. YSX or appropriate derivatives thereof may be condensed. Alternatively, if an in-phase method is used, 9-fluorenylmethyloxycarbonyl-aminoacyl-p-alkoxybenzyl alcohol resin is used as the starting material and 9- Removal of fluorenylmethyloxycarbonyl (FmOC) group, 1”
It can be synthesized by sequentially repeating the synthesis of moc-amino acids and finally condensing benzyloxycarbonyl-amino acids, Boc-amino acids, or carboxylic acids.

In the general formula [■1, when X' and Y' contain an amino acid containing a water 11f group such as serine or threonine, and when trifluoroacetic acid is used for final deprotection in the in-phase method or liquid phase method, each The amino acid may be protected as t-butyl ether. Furthermore, when catalytic hydrogenolysis is used for final deprotection in the liquid phase method, each amino acid may be protected as a benzyl ether.

The "amino acids" appearing in the definitions of X and Y or the amino acids constituting peptide 1 include α-amino acids, but they do not necessarily have to be α-amino acids.

As α-amino acids, the following formula % formula % [wherein R2 is (1) hydrogen, (2) unsubstituted lower alkyl, or (3) substituted lower alkyl (II substituents include amino group, hydroxy, mercapto, lower (alkylthio, carboxylphenyl, human Oxyphenyl, imidazole group, indolyl group, etc.). ) or an amino acid represented by 2H (in the formula, R3 represents hydrogen or a hydroxy group).

Specifically, these amino acids include glycine, alanine, valineleucine, isoleucine, serine, threonine, cysteine, methionine, aspartic acid, glutamic acid, lysine, arginine, phenylalanine, tyrosine, histidine, tryptophan, proline, oxyproline, etc. can give.

Examples of peptides include generally low-polymerized peptides in which about 2 to 3 of the above amino acids are bonded.

In addition, as a protecting group for the amino group in X etc., (1)
Acyl groups, including phthalyl groups and sulfonyl groups, and (
2) Examples include oxycarbonyl groups that are protected in the form of urethane. For example, the acyl group in (1) is an unsubstituted lower acyl group. Examples include alkylcarbonyl, substituted lower alkylcarbonyl (substituents include halogen atom, nitro, lower alkoxy, )I-yl, etc.), benzoyl, phthalylarylsulfonyl, and the like. These acyl groups having a phenyl group may have a substituent on the phenyl group, such as lower alkyl, halogen atom, lower alkoxy, or nitro.

Specific examples of these include acetyl, trifluoroacetyl, phenylacetyl, phenylbutanoyl, phenylpropionyl, benzoyl, and the like.

The oxycarboxy group to be urethanized and retained in (2) is unsubstituted lower flukoxycarbonyl, substituted lower alkyloxycarbonyl (the substituent is the one described in (1) above).
The same examples are given for the acyl group.

) can be given. Specific examples include isopropyloxycarbonyl, t-butoxycarbonyl, impendeloxycarbonyl, and benzyloxycarbonyl. ) etc.

In addition to the protective groups (1) and (2) above, benzyl, substituted benzyl (the substituents include the same ones as above),
O-nitrophinyldio, triphenylmethylthio, and triphenylmethylthio can be used.

Examples of the saturated or unsaturated hydrocarbon group in R1 (2) include saturated or unsaturated lower aliphatic hydrocarbon groups and aromatic hydrocarbon groups. These may have substituents. Specific examples of these include lower alkyl groups, benzyl groups, and phenyl groups.

Examples of esters for Y include esters with lower alcohols, benzyl alcohol, and the like.

In the present invention, examples of lower alkyl include C1 to C5 furkyl such as methyl, ethyl, propyl, butyl, and pentyl.

Examples of lower alcohols include C1 to C5 alcohols such as methanol, ethanol, propatool, butanol, and pentyl alcohol.

In the general formula [V], preferably R is methyl, ethyl, propyl, benzyl,
Examples include peptides in which any of these amino acids such as phenylbutanoyl-valine, benzyloxycarbonyl-aspartic acid, and benzyloxycarbonyl-lysine are condensed with amino acids such as valine, phenylalanine, proline, or threonine.

Y includes D-leucyl-valine, glycyl-valine, di-leucyl-valine and esters thereof, such as lower fulkyl esters such as methyl or t-butyl ester, or benzyl varnish. The structure, physicochemical properties, and endobebutidase inhibitory activity of the endobebutidase inhibitory peptide produced according to the present invention are summarized in Table 1.

Family prolyl endopeptidase activity was determined using the method of Yoslwoto et al. [(8iochim, Pi
phys, Acta), Vol. 569, 184-192 (1979)]. That is, 40 in a test tube
0.518 N-benzyloxycarbonyl-di-glycyl-prolyl-β-naphthylamide containing % dioxane, 0.2e, 0.05M Tris-salt 1m! lii
Liquid (+) H7,8) 0.5ae, aqueous solution containing specimen 0
．． 251! After heating the mixed solution to 37 degrees for 3 minutes,
1. CheIm, vol. 1251, 7593-7599
This is an enzyme purification method based on an improvement of the method described in Page (1976), using ammonium sulfate fractionation from pig kidneys, DE
Prolyl endopeptidase solution purified by AE-Sephadex A-50 column chromatography.
05d was added and reacted at 37 degrees for 30 minutes. After the reaction, 1M sodium acetate (pH 4,2) IIl containing 10% of Tween Bween-20) and 0.1% of First Garnet GBC@ was added, and after being left at temperature V for 1 b, the absorbance at 525 nl was measured. (a) was measured. At the same time, the absorbance (b) of a blind reaction solution using only the mild WjJ solution containing no specimen was measured, and the prolyl endopeptidase inhibition rate was calculated from the formula [(ba)/b]×100.

Anti-elastapi inhibitory activity was determined by HLIIlfOrd, RA
[J, Biol, Chet vol. 225, 2
227 (1980)], the anti-tebusin B inhibitory activity is K
11i (lhtCG, method [Biochew, J.
189, p. 447453 (1980) 1.

f1 Rf3 f4 Rf2 =TLC Reverse Phase
(RP-18) KOAC 5%-Quenll 1%:H
eCN=65235-TLC, silica gel 1゜n-BuOH:AcOH:H2O=4:1:1=T
LC, silica gel F.

CHCl3:He0H=^cOH= 90:10:5P
CE 1ast at-a Boc B P A z OBz l 0 But 0 He = Prolyl endopeptidase inhibitory activity, IC50 (u9/me) 11 Lastase inhibitory activity.

ICAO (u9/me) Single tepsin B inhibitory activity IC50 (u9/me) #Benzyloxycarbonyl = t-butoxycarbonyl = phenylbutanoyl = phenylpropionyl = phenylacetyl monobenzoyl = benzyl ester - t-butyl ester monomethyl ester [Example 1] The present invention will be specifically explained below with reference to Examples, but the present invention is not limited thereto.

Example 1 Synthesis of Boc-D-isoyl-L-valine benzyl ester 0C-D-Cl ysine monohydrate 0.7499 (3,
OOsmol), L-valine benzyl ester p-
Toluene sulfonate 1.2529 (3, 30
1101) and N-human Oxylbenzotriazole 0
．． 8 1 1 9 (6.
Ethyl-3-(3-dimethylaminopropyl)carbodiimide perchlorate 1.108g (4.20tl
) and stirred for 2 hours, then roughly stirred at room temperature for 4 hours.

After the reaction was completed, 30e of water and 15d of a 1% aqueous citric acid solution were added, and the precipitated crystals were filtered and separated. Dip the organic layer into 20 m of water.
, Washed with 20 ml of saturated aqueous sodium bicarbonate solution and 10 d of water, then anhydrous! After drying the lI-coated sodium chloride solution, the solvent was distilled off to obtain 1.39 CI of crude Boc-D-leucyl-monovaline benzyl ester as white crystals. Next, the crude crystals were purified by column chromatography (silica gel, ethyl acetate:hexane (5:2)) to obtain 1.233 g of BOC-D-leucyl-shi-valine benzyl ester as white crystals. Yield 97.8%.

1982.0-83.0℃.

[α] D+35.1' [α] +76, 6.

(cl.02, chloroform).

1) 1-NMR (CDC1 3) δ 0.85.0.92 (d, d.3H.3H.

J=7Hz. CH3') 0, 90-0.98 (d, d, 3H.3H.

J = 2, 7 Hz, C H3) 1, 3
6-1.54 (m, IOH.

(CH3)3COCO, (CH3)2 CH - CH a H b )
1, 62-1.76 (m, 2H.

(CH3)2CH-CHaHb) 2. 14-2. 27 (m, IH.

(CH3)2CHCH) 4, 14 (br. 1H. NHCHcO
NI-1) 4, 57 (dd. 1) (, J=
9. 2.

4, 2) 1z. CONMCI-1cOO)4,
79 (br. 1H.NH>5, 13.
5. 19 (d, d, IH. IH.

J-1 2.0Hz. CI-12Pl"t) 6, 68
(br, IH, NH) 7,29-7.40 (m.5H.Ph) Example 2 In Example 1, N-human O-oxysuccinimide 0,6939 (6 ．．
Boc-D-0 isyl-L-valine benzyl ester 0.71057 was obtained in the same manner.

Example 3 Synthesis of threo(2R.38), (28.3R)-3-(p-methoxybenzyloxycarbonyl)amino-2-hydroxypentanoic acid threo(2R,3S), (2S,3R)-3- Amino
2-Hydroxypentane 115.999 (45,0s■
01) with 8-4.6-dimethylpyrimidin-2-ylthiolcarbobenzyl 15. 07SF (49.5g+nol
), 25 m of water, 25 m of dioxane, and 9,45 ae (67.5 veol) of triethylamine were added, and the mixture was stirred at room temperature for 24 hours. After the reaction was completed, 65 ae of water was added, and after washing twice with 100 d of ethyl acetate, the aqueous layer was cooled in a water bath. The aqueous layer was adjusted to pH 2 by adding 5N hydrochloric acid (22-mL), and then extracted once with 65 d of ethyl acetate and twice with 30 M1. The oil layer was diluted with 50 ae of 5% hydrochloric acid three times and 50 ae of saturated saline.
After washing twice with

3S), (28.3R)-3-(p-methoxybenzyloxycarbonyl)amino-2-hydroxypentanoic acid 9.6591 was obtained. Yield 72.1%.

Example 4 Elito0 (2R.38), (28,3R)-3-(p
Synthesis of -methoxybenzyloxycarbonyl)amino-2-hydroxypentanoic acid Threo (2R, 38), (28) of Example 3.

3R)-3-amino-2-hydroxypentanoic acid instead of erythro (2R.38), (28.

3R)-3-amino-2-hydroxypentanoic acid 5,
Using 99SF (45. Ollol), 110.769 of erythro (2R.38), (28.3R)-3(p-methoxybenzyloxycarbonyl)amino-2-hydroxypentane was obtained in the same manner.

Example 5 Z (OMe)- (2R.38)-3-amino-2-
Synthesis of hydroxypentanoyl-D-leucyl-d-valine benzyl ester 3oc-D-leucyl-d-valine benzyl ester 1
89.2N (0.450 msol) was added with 2 m of trifluoroacetic acid, and after stirring at room temperature for 40 minutes, the solvent was distilled off.
Further, add 2I11 of toluene and distill off the solvent.
By repeating this process twice, D-leucyl-shibaline benzyl ester trifluoroacetate was obtained as white crystals. Here, (2R.38) 3-(p-methoxybenzyloxycarbonylamino-2-human oxypentane 1133.
811g (0.45011o1), N-hydroxybenzotriazole 91, 2Q (0.67
5gvol) and DMF 2ae were added and dissolved, and N-methylmorpholine 50.4μ! was added under water cooling. (0.4
50 mmol) and 97% 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide perchlorate 1
48. Add 3ae (0.563gimol) and add 2
After stirring for an hour, the mixture was further stirred at room temperature overnight. After the reaction is complete, the reaction mixture is diluted with 30 m of dichloromethane, washed sequentially with 10 m of a 1% aqueous citric acid solution, 10 m of water, 10 m of a saturated aqueous sodium bicarbonate solution, and 10 m of water, dried over anhydrous sodium sulfate, and then the solvent is distilled off. Thus, the oily crude Z (OMe)- (2R.38)-3-amino-2-
Hydroxypentanoyl-D-leucyl-di-valine benzyl ester 316, 5* was obtained. Next, this assembly was purified by column chromatography (silica gel, ethyl acetate:hexane (5:1)) to obtain Z (OMe)- (2R.38)-3-7 minnow 2 as white crystals.
-Human O-oxypentanoyl-leucyl-L-valine benzyl ester 197.6 µm was obtained. Yield 73.2%
. ■p115.0~115.5102, 9' (cl
．． 04, chloroform).

1 H-NMR (CDC1 3) 6 0, 80-1.04 (m.1 5H.CH3×5
) 1, 51-1. 80 (m, 5
G1. CH 2x2. CH (Leu)
) 2, 19 (m.1H.CH (Val)) 3, 7
0-3.85 (m.4H. CH30.

CH (EtAhp)) 4, 12 (dd, IH.J=6.4.

3.0Hz. CHOH) 4, 52 (dd.1H.J=8.4.

4,8Hz. CH(Val))4.49
(m, IH, CH(Leu))4.61 (br
d, IH, J=6.4Hz, 0H) 4.98 (s, 2H, CH2-C6H4) 5.08,
5.18 (d, d, 2H.

J=12.4Hz, CH2Ph) 5.31 (br d', IH, J=8, 4
t1z, NH(EtAhp) )6.79
(br d, IH, J= 8.4Hz, NH(Val)) 6.84-6.92 (m, 2H, C6H4) 7.07
(br d, IH, J= 8.4Hz, NH(Leu)) 7.22-7.40 (m, 7H, C6H4゜Ph) However, EtAhp is 3-amino-2-hydroxypentanoic acid Example 6 Boc- L-t<+) Leu (2R,3S)-3-7 minnow 2-human Oxypentanoyl D-leucyl
Synthesis of valine benzyl ester Z (OMe)-(2R,3S)-3-amino-2-hydroxypentanoyl-D-leucyl-L-valine benzyl ester 190.2ay (0,317--01) with anisole O, Add 1m and 2d of trifluoroacetic acid,
After stirring at room temperature for 40 minutes, the solvent was distilled off, and toluene
By adding 〆 and distilling off the solvent twice, (2R,3S)-D-leucyl-valine benzyl ester trifluoroacetate was obtained as white amorphous.243.6
I got ■. To 219.2 μg (equivalent to 0,285 smol) of this salt was added 65.4 II g (0,301 mol) of BOC-L-valine.
Peng mol), N-hydroxybenzotriazole 5
8.1■ (0,430mg1ol) and DMF 2a
Add and dissolve N-methylmorpholine 32 under water cooling.
μm (0,286 wmol) and 97% 1-ethyl-
3-(3-dimethylamitube 0vir)carbodiimide.
After adding 94.0 μl (0,357 μl) of perchlorate and stirring for 2 hours, the mixture was further stirred at room temperature overnight. After the reaction is complete,
The reaction mixture was diluted with 30 d of dichloromethane and 10 d of 1% aqueous citric acid solution! , Wednesday 10ad! , sequentially washed with 10 Id of saturated aqueous sodium bicarbonate solution and 10 m of water, dried with sodium viate anhydride, and then distilled off the solvent to obtain an oily crude Boc-L-valyl-(2R13S)-3-amino-
520.8 ml of 2-hydroxypentanoyl-Qoyl-valine benzyl ester was obtained.

The mouth assembly was then purified by column chromatography (silica gel, dichloromethane:methanol (24:1)).
By doing so, Boc-L-valyl-(2
R,38)-3-Amino-2-human Oxypentanoyl D-leucyl-L-valine benzyl ester 149.
I got 7■. Yield 82.8%. It) 167.0-16
8.081.0° (C1,00, chloroform).

1 H-NMR (CDCI 3) 60.80-1.03 (m, 21H, CH3X7) 1, 44 (S, 91-1° (CH3)3COCO) 1.52-1.90 (m, 5) 1 ．． CH2x2, CH
(Leu)) 2.11 (m, IH, CH(Vat)>2.19(
m, 1H, C)-1(Val)>3, 87 (m,
1H.

CHNH(EtAhp)) 3.93 (dd, 1H, J=5.8°8.5Hz,
CH(Val))4.10 (dd, IF(
, J=3.4°7.0) (z, C)-108) 4.36 (m, It-1, Ct-1(Leu))4.
58 (dd, IH, J=5.0°8.6Hz, CH
(Val)) 5.09 (br, IH, NH(Val))5.
10,5.20 (d, d, 2H, J=12.2Hz
, CH2Ph) 5.36 (br, 1H, OH) 6.98 (br d, 1H, J=8.6H
z, NH(Va I )) 7.04 (br
d, IH, J = 8.4Hz, NH (EtAhp)) 7.20 (br d, 1H, J = 8.4Hz, NH (Leu) > 7.30-7.40 (m, 5H, Ph) However EtA
hp is 3-amino-2-hydroxypentanoic acid Example 7 Z-L-valyl-shi-valyl-(2R,3S)-3-amino-2-hydroxypentanoyl-D-〇isyl L
-Synthesis of valine benzyl ester 0C-L-valyl-
(2R,38)-3-Amino-2-human Oxypentanoyl D-leucyl-shi-valine benzyl ester 14
Add 2IIIl of trifluoroacetic acid to 5.0IIg (0,228--01), stir at room temperature for 40 minutes, evaporate the solvent, add 2d of toluene, and evaporate the solvent.
By repeating this process twice, white crystals are obtained as L-valyl (2R93
3)-3-amino-2-hydroxypentanoyl-〇-
Isyl-[-valine benzyl ester trifluoroacetate salt was obtained. This salt was diluted to 10 ae with ethyl acetate and 1 ml was added! The solvent remaining after sampling was distilled off (equivalent to 0.2061101).

Here, Z-L-valine 54.8■ (0,218+uol
), N-hydroxybenzotriazole 42, 4j
FF (0,314*mol) and DMFi, 5IIi
Add and dissolve 23μ of N-methylmorpholine under water cooling.
m (0,206 smol) and 97% 1-ethyl-3
-(3-dimethylaminopropyl)carbodiimide perchlorate 68.0Mg (0,258wnol) was added to 2
After stirring for an hour, the mixture was further stirred at room temperature overnight. After the reaction was completed, the reaction mixture was diluted with 15 ae of dichloromethane and diluted with 1%
By sequentially washing with 7 ml of citric acid aqueous solution, 7 d of water, 7 d of saturated sodium bicarbonate aqueous solution, and 7 d of saturated brine, and drying with anhydrous sodium sulfate, the solvent was distilled off to obtain crude Z-L-valyl as partial crystal amorphous. Bariloo (2R
,3S)-3-amino-2hydroxypentanoyl-〇-leucyl-valine benzyl ester 277.9■
I got it. This phase was then purified by column chromatography (silica gel, dichloromethane:methanol (40
:1)) As a white crystal, Z-shi-baryl-
L-valyl-(2R,38)-3-amino-2-human O
125.9 ml of xypentanoyl-D-leucyl-d-valine benzyl ester was obtained. Yield 79.7%.

11) 222.0-223.0°C. [α], +9.2°
[α] +24.4° (C1,01, on
Holm), Si-MS m/z 76B (M+1
+), 790 (M+Na+).

1)-1-NMR (CDCI 3) 60.76-1.00 (m, 271-1.CH3X9
) 1.37 (m, 1H, (CH3-CHaHb) 1.53-1.85 (m, 4H.

(CH) CHCH, CH3 HaHb) 1.93-2. 10 (m, 2H, CHX2
(Val) ) 2.17 (m, IH, CH (Vat)) 4.13-4.
27 (m, 2H.

HNCHCH(OH)Go) 4.41-4.64 (m, 4H, CHx3(Va
l), CH(Leu)) 5.06.5.15 (d, d, 2H.

J=12.0Hz, C00CH2Ph)5.06.5.
09 (d, d, 2H.

J=12.0Hz, PhCH2000) 5.49 (
br s, 1)-1,0H) 5.86 (br, I
H, NH(Val)) 7.09 (br, 1
day, NH (Va I)) 7.22 (br, 1H,
NH (EtAM)) 7.22-7.39 (m, 10H.

Phx2) 7.45 (br, d, 1H, J= 7.8Hz, NH (Leu) 7.94-8.23 (br, 1H.

NH(Val)) However, EtAhp is 3-amino-2-hydroxypentanoic acid Example 8 Z-L-valyl-shi-valyl-(38)-3-amino-
Synthesis of 2-oxopentanoyl-D-leucyl-shi-valine benzyl ester Z-L-valine-L-valyl-(2R,38)-3-amino-2-hydroxypentanoyl-D-leucyl-L-
Valine benzyl ester 108, ill (0,14
1Smol) Nibilicin trifluoroacetate 19.
9■ (0,103s103s, 97% 1-ethyl-3-
<3-dimethylaminopropyl)carbodiimide perchlorate 116.2q (0,441Smol), DMS
Add O0,5ae and benzene 2IIIl and stir at room temperature for 24
Stir for hours.

After the reaction was completed, the reaction mixture was diluted with ethyl acetate 15e,
After washing with 5 m of water, drying with anhydrous sodium sulfate, and distilling off the solvent, a partially gel-like crude ZL-valyl-L-valyl-(38)-3-amino-2-oxopentanoyl-D was obtained. -Leucyl-d-valine benzyl ester was obtained.

This assembly was then purified by 11-layer chromatography (silica gel, black methane:methanol (15:1)) to obtain Z-L-valyl-valyl-(38)-3-amino as white crystals. -2-oxopentanoyl-D-leucyl-d-valine benzyl ester 63.2
I got ay. Yield 58.6%.

1D201. o~202.0°C. [α], +8.4°
[α] +50.3° (co, 90, chloroform).

1H-NMR (CDC13) δ 0.77 1.04 (m, 27H, CH3X9) 1.50-1.85 (m, 4H.

CHCH, CH3-Cl-1aHb) 2.00 (m, IH, CH3-CHaHb) 2.05
-2.25 (m, 3H, CHx3(■al)) 4.06 (m, 1H, CH(Val)) 4.32
(dd, 1H, J=7.4Hz.

8.6) 1z, Cl-1(Val))4.47-4
．． 63 (m, 2) (.

CH(Val), C)-1(Leu))5, 0
8. 5. 16 (d, d, 2
1.

J=12.0Hz, CH2-Ph) 5.10.5.11 (d, (1,2H.

CH2-Ph) 5.34 (ddd, IH, J=4.8°8.2.8.
2Hz.

CH(EtAOl))) 5, 57 (br, 6. 1H.

J=8.6Hz, NH(Val))6.69 (b
r, d, 1H.

J-8,6Hz,NH(Val))6.74(b
r, d, IH.

J=9.1 Hz, NH(Va I ))6.86 (
br, d, IH.

J=8.2Hz, NH(EtAho))7.25-7.
40 (m, IOH.

phx2) 7.48 (br　d, IH.

J=8.8Hz, NH(Leu)) However, EtAhp is 3-amino-2-hydroxypentanoic acid Example 9 Poststatin (shi-valyl-L-valyl-(3S)-
Synthesis of 3-amino-2-oxopentanoyl-D-leucyl-valine) Z -L-ha + Jru L-ha + Jru (3S)
-3-Amino-2-oxopentanoyl-〇-leucyl-1-valine benzyl ester 46.9q (0,061
Palladium black 6.6 and methanol 12- were added to the mixture (g+gtol), and after purging the system with hydrogen, catalytic reduction was carried out at room temperature for 20.5 hours. After removing the catalyst by filtration, the solvent of the filtrate was distilled off to obtain 40.0 cm of white crystals. Of this, 39.7q was analyzed by HPLC (column: CAPCELL PAK C1
8,5HISEIDO, 20X250am, i After leaving:
1% citric acid, 0.65% potassium acetate aqueous solution niacetonitrile = 8:2, flow rate: 8ai! /sin, detection wavelength: 25411), the unreacted raw material with the target product and a portion of the raw material produced by ebimerization are valyl-(3R)-3-amino-2-hydroxypentanoyl-D- Separate isyl-L-valine and
A fraction containing L-valyl-(38)-3-amino-2-hydroxypentanoyl-D-leucyl-shi-valine was collected.

After concentrating this fraction at 25°C, the concentrated liquid was transferred to CHP2
0 resin, washed with water, and eluted with 80% methanol. The eluate was evaporated and dried overnight using a vacuum pump to obtain 15.2 ay of poststatin as a white powder. Since this poststatin contained impurities derived from CHP20 resin by NMR, 5.1Mg of this was applied to CHP20 resin again and eluted with 55VO1% methanol. After the eluate was distilled off, the resulting solid was decanted. By washing with ether in a vacuum pump and drying with a vacuum pump, Poststatin 2.4■ was obtained as a white powder. Yield 21.7%. 81-MS m/z542 (M+1+
).

1H-NMR (DMSO-d6) δ 0. 70 0.95 (m, 27 H, CH
3x9) 1.40-1.67 (m, 3H.

CHaHbCH(Leu), CH2 OHa　Hb　) 1, 61 (m, IH.

Cl-1aHbcH(Leu)) 1.76 (m, 1H, CH3-C)-1aHb)1.
86-2. 10 (m, 3F1°CH (Va
l )) 3.20 (m, I H, CH(Va l ))
4.06 (m, 1H, CH (Val)) 4, 27
(m, 1H,CH(VaI))4.4
3 (m, IH, CH (Leu)) 4.95
(m, IH, C)-1cOco)7.98(br
d, IH.

J=8.4Hz, NH(Val))8.10 (
br d, 1) 1°J=8.9Hz, NH(Va
l)>8.36 (d, IH, J-6, 6Hz.

NH(EtAhp)') 8.56 (d, 1H, J=8.7Hz.

NH (Leu)) However, EtAhp has a broad peak derived from 3-amino-2-hydroxypentanoic acid (contains 7 amino groups and carboxyl groups at 62.80-3.90).

Example 10 Synthesis of Z-D-o Isil-L-valine t-butyl ester Z-D-o Isil>1.0009 (3,77-■01)
, L-valine t-butyl ester hydrochloride 0,719
(3,43e+mol) and N-hydroxylbensito IJ 7sol 0.9279 (6,00+nol)
was dissolved in 0.530ad of triethylamine and cooled with water. (3,79gimol) and 97% 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide perchlorate 1.2659 (4,8
After stirring for 2 hours, the mixture was further stirred at room temperature for 14 hours. After completion of the reaction, 7 ae of saturated aqueous sodium bicarbonate solution, 7 yd of water, 7 ae of 1% citric acid aqueous solution and 7 ae of water
The mixture was washed successively with MN, dried over anhydrous sodium sulfate, and then the solvent was distilled off to obtain crude Z-D-leucyl-monovaline 1-butyl ester 1.9329 in the form of an oil. Next, the crude crystals were purified by column chromatography (silica gel, methane:methanol (400:3)') to give Z- as white crystals.
D-0 Isil-L-valine t-butyl ester 1.26
I got 79.

(C1,06, Rinoform).

Example 11 Eris0 (2R,3R), (28,33)-Z(OM
e) -3-amino-2-human O-oxypentanoyl〇-
Synthesis of leucyl-l-valine t-butyl ester Z-D-leucyl-L-valine t-butyl ester 50
3.9 mg (1,198-mol) of palladium black 21
．． After adding 2q and 5d of methanol and replacing the system with hydrogen,
Catalytic reduction was carried out at room temperature for 3 hours. After removing the catalyst by filtration, the filtrate was distilled off to obtain oily D-isoyl-L-valine t-
Butyl ester was obtained. Here, Erythro (2R, 3R
), (28°33)-3-(p-methoxybendiJreoxycarbonyl)amino-2-hydroxypentanoic acid 3
91.8η (1,318 I of), 324.1 of N-hydroxylbenzotriazole (2,399 ssol) and 8 m of dichloromethane were added and dissolved, and 97% 1-ethyl-3-(3-dimethylamino) was dissolved under water cooling. Propyl) Carbodiimide/Perchlorate 44
After adding 2.1■ (1,677 dew-01) and stirring for 2 hours,
It was further stirred at room temperature overnight.

After the reaction was completed, the reaction mixture was mixed with 8M1 of a saturated aqueous sodium bicarbonate solution, 8D of water, and 817M of a 1% aqueous citric acid solution. After washing with 8 ae of water and drying with anhydrous sodium sulfate, solution I! By distilling off, a candy-like crude 1 Lith 0 (2R, 3R
), (28,3S)-Z(OMe)3-amino-2-hydroxypentanoyl-D-acyl-[-valine t-
Butyl 1 ster 72B, 1■ was obtained. Next, this assembly was purified by column chromatography (silica gel, dichloromethane:methanol (100:1)) to obtain erythro (2R, 3R) as an amorphous solid.

(28,38)-Z (OMo)-3-amino-2-hydroxypentanoyl-D-leucyl-shi-valine t-
Butyl ester 666.9191 was obtained.

Yield 98.4%. Sl-M5 m/z 566 (M
+1+).

Example 12 Synthesis of Z-L-phenylalanyl<2R8,3R8)-3-amino-2-hydroxypentanoyl-D-leucyl-shi-valine t-butyl ester Eris0(2R,3
R), (28,38)-Z(OMe)-3-F mino-2
-Hydroxypentanoyl-D-leucyl-L-valine t-butyl ester 340.94 (0,603 ssol
) in 4 m of methanol, palladium black 50.7 j
19 was added in three portions every few hours, and the mixture was exposed to contact light for 32 hours in a hydrogen atmosphere in a V-lagoon. After removing the catalyst by filtration, the filtrate was distilled off to obtain Erythro (2R93R), (28
,38)-3-amino-2-hydroxypentanoyl-
D-0 isyl-L-valine t-butyl ester was obtained.

Z-L-phenylalanine 100.
8 m (0.337 uiol) and 10 d of N-hydroxybenzotriazole chloromethane were added, and 97% 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide perchlorate 111.3 jl (0.4 211o
After adding 1) and stirring for 2 hours, the mixture was further stirred at room temperature for 18 hours. DMF 2. After adding Oae and stirring for an additional 22.5 hours, the reaction mixture was diluted with a saturated aqueous sodium bicarbonate solution.
d, water 9ai! , 9d of 1% citric acid aqueous solution and 9d of water, dried over anhydrous sodium sulfate, and the solvent was distilled off to obtain oily crude Z-L-phenylalanyl-(2R
S.

3RS)-3-amino-2-hydroxypentanoyl-
〇-Leucyl-1-valine-t-butyl ester 255.
I got 619. This assembly was used in the next reaction without purification.

Example 13 Z-L-7enylalanyl(RS)-3-amino-2
-oxopentanoyl-[)-0isyl[-valinet
Synthesis of -butyl ester Crude Z-L-phenylalanyl-(
2RS,3RS)-3-amino-2-hydroxypentanoyl-D-leucyl-shi-valine benzyl ester 2
55. Pyridine trifluoroacetate 25.6 ay.
8jlg (0.1 341101), 97% 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide perchlorate 211.6q (0.803veo
l), DMSO 0. 65ae and benzene 2IIIi were added, and the mixture was stirred at room temperature for 20 hours.

After the reaction is complete, the yellow reaction mixture is diluted with 6,57 ml of ethyl acetate and 77 ml of water. After drying with anhydrous sodium sulfate, the amorphous solid Z-1-
244.4 ml of phenylalanyl-(R8)=3-amino-2-oxopentanoyl-D-leucyl-valine t-butyl ester was obtained. Next, this assembly was purified by column chromatography (silica gel, dichloromethane:methanol (100:1)) to obtain an amorphous solid Z-L-phenylalanyl-(R3)-3.
-Amino-2-oxopentanoyl-D-leucyl-shi-valine t-butyl ester 113.7II! I got g. Yield 63.5%. 31M5 m/z681 (M+
1+).

Example 14 Z-L-phenylalanyl-(R8)-3-amino-2
-Synthesis of oxopentanoyl-D-D isyl-L-valine Z-L-7enylalanyl-(R8)-3-amino-2
-oxopentanoyl-[)-0isyl-[-valinet
-Butyl ester 105.9■ (0,156gH1ol
) and stirred at room temperature for 90 minutes, the solvent was distilled off, and toluene 2111e was added and the solvent was distilled off twice to obtain Z-L-phenylalanyl as an amorphous solid. 106.511 g of -(R8)-3-amino-2-oxopentanoyl-D-leucyl-d-valine was obtained. Next, this assembly was purified by column chromatography (LH-20, methanol) to obtain Z-L-phenylalanyl-(R8)-3-amino-2-oxopentanoyl-D- as amorphous solid white crystals. 70.5 Mg of leucyl-valine was obtained. Yield 72.6%.

81-M5 m/z 625 (M+1+), 647
(M+Na+).

Example 15 Z (OMe)-(2R8,3R8)-3-ami/2
-Synthesis of hydroxypentanoyl-Q-0 isine t-butyl ester - Leucine t-butyl ester salt 1 salt 0.896g
(4,001aol) to (2R8゜3R8)-3-(p
-methoxybenzyloxycarbonyl)amino-2-hydroxypentanoic acid 1.3099 (4,40■-of
), N-hydroxybenzotriazole 1.081
9 (8,001101) and dichloromethane 2C)d
! Add 0.62 m of triethyl chloride under water cooling.
(4,43 mmol) and 97% 1-ethyl-3-(
1.476 SF (5,6011 o1) of 3-dimethylaminopropyl)carbodiimide perchlorate was added, and after stirring for 2 hours, the mixture was further stirred at room temperature for 15 hours. After the reaction is complete, the reaction mixture is mixed with 8e of saturated aqueous sodium bicarbonate solution and 8a1 of water. , washed sequentially with 1% aqueous citric acid solution 8e and water 8d, dried over anhydrous sodium sulfate, and distilled off the solvent to obtain candy-like crude Z(OMe)-(2R8,3R8)-3.
-Amino-2-hydroxypentanoyl-D-leucine t-butyl ester 2.1229 was obtained. Next, this assembly was purified by column chromatography (silica gel, dichloromethane:methanol (100:1)) to obtain Z (OMe)-(2R3,3R3) as white crystals.
-3-7 minnow 2-hydroxypentanoyl-D-leucine t-butyl ester 1.8589 was obtained. Yield 99
．． 6%. 81-M5 m/Z467 (M+1+).

Example 16 BOC-L-valyl-(2R8,3R8)-3amino-
Synthesis of 2-hydroxypentanoyl-D-leucine t-butyl ester Erythro(2R,3R), (28,38>-Z(OMe
)-3-amino-2-hydroxypentanoyl-D-0
Isin t-butyl ester 303.9■ (0,651s
sol) in methanol 4d, palladium black 43
．． 4Q was added in three portions every few hours, and catalytic reduction was carried out at room temperature in a hydrogen atmosphere for 15 hours. After removing the catalyst by filtration, the filtrate was distilled off to obtain Eris 0 (2R, 3R).

(28,38)-3-amino-2-hydroxypentanoyl-D-leucine t-butyl ester was obtained. Here, Boc-L-valine 155.9m (0,718m5ol
), N-hydro-mainly sylbenzotriazole 176.1
■ (1,303 mmol) and dichloromethane 3.5 m
and 1-ethyl-3-(3-dimethylamitube Ovir)carbodiimide hydrochloride 499.4j1 under water cooling.
9 (2,605■■01) was added and stirred for 2 hours, followed by further stirring at room temperature for 16.5 hours. After diluting the reaction mixture by adding 7IIl of dichloromethane, 6d of 1% citric acid aqueous solution was added.
, water 6111, saturated hydrogen carbonate ± thorium aqueous solution 6I11
and water 6-, dried over anhydrous sodium sulfate, and distilled off the solvent to obtain amorphous crude Boc-L.
-valyl-(2R3,3R8)-3amino-2-human0
Xypentanoyl-D-leucine t-butyl ester was obtained. This assembly was used in the next reaction without purification.

Example 17 Synthesis of E3oc-L-valyl-(R8)-3-amino-2-oxbentanoyl-o-o-isyl t-butyl ester oc-L-valyl-(2R8,3R3)-3amino-
69.59 (0,360-mol) of pyridine trifluoroacetate, 1.3ae of dicyclohexylcarbodiimide DMSO and 2.5d of benzene were added to 0.651 mmol of 2-hydroxypentanoyl-〇-leucyl t-butyl Nisder,
Stirred at room temperature for 18 hours. After the reaction was completed, the yellow reaction mixture was diluted with 10 ml of ethyl acetate, and the DCC urea produced was filtered off. The organic layer was washed with 10 ml of water, dried over anhydrous sodium sulfate, and then the solvent was distilled off. 5 ml of ethyl acetate was added again to the obtained oil, the precipitated DCC urea was filtered off, and the filtrate was distilled off to give crude 3oc-L-valyl-
(RS)-3amino-2-oxopentanoyl-D-leucyl t-butyl ester-L-valine benzyl ester was obtained. Next, this assembly was subjected to column chromatography (silica gel, dichloromethane:methanol (10
0:1), ■ Diku on methane: ethyl vinegar (12:1~
10:1), Boc-
178.9 ml of L-valyl-(RS) 3-amino-2-oxopentanoyl-D-leucyl t-butyl ester was obtained.

31-MS m/z 534 (M+1+-).

Example 18 Synthesis of (2R.38)-3-(9-fluorenylmethyloxycarbonyltanic acid) 0.7599 (5.70 gisol) of -3-amino-2-hydroxypentanoic acid was added with carbonic acid. An aqueous solution of sodium hydrogen (1.1999 <14.28 ssol) dissolved in 20 ae of water and 20 d of dioxane were added, and 9-fluorenylmethyl O-roformate (1.62 ssol) was added under ice cooling.
39 (6.27 ssol) dissolved in dioxane 10- was added dropwise over 30 minutes. After the completion of the dropwise addition, the mixture was stirred at room temperature for 2 hours, and then water 100111e was added to dilute the reaction mixture, and the mixture was washed once with ether 150- and 75I11. Add 2.8-6N hydrochloric acid to the aqueous layer. 5, and extracted three times with 50 Ill of ethyl acetate. The oil layer was washed with 50 m of saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off to obtain crude (2R.3S)-3-(9-fluorenylmethyloxycarbonylcypentanoic acid 1.6229). After adding ethyl acetate 4se and n-hexane 4Id to this assembly and causing crystallization, the crystals were collected by filtration and mixed with ethyl acetate-n-hexane (1:1).
After washing with solution 2d and drying, a white solid (2R.
38)-3-(9-Fluorenylmethyloxycarbonyl)amino-2-hydroxypentanoic acid 0.99(i
I got s. mp160-161.5°C. Then, this N
- Add 4ae of ethyl acetate to 0.9999 of the protected amino acid and bath a! After stirring for 40 minutes while suspending at 50°C,
- 4 d of hexane was added and the crystals were collected by filtration. After washing the crystals with ethyl acetate-n-hexane (1:1) solution 2- and drying, white crystals of (2R.38) 3-(9-fluorenylmethyloxycarbonyl)amino-2-hydroxypentane were obtained. Acid (cl.01.methanol). 2626mp
162-162.5°C. FAB-MSm/z
356 (M+1+).

1 H-NMR (CDC13) 6 0.96 (dd, 3H, J=7.4°7.4H
z, 0H3) 1.69 (m, 2H, CH2) 3, 25 (br, C0OH, 0H) 4.01
(m, IH, CHNH) 4, 18 (dd, 1H, J=6.6゜6.6
Hz, ArCHAr) 4, 24 (br s, 1H, CHOH)
4.40.4.41 (d, d, IH, IH.

J-6,6,6,6H2゜ ArCHCHHaHb) 5.13 (d, 1)-1, J=9.6Hz.

NH) 7.20-7.80 (m, 8H, Ar) On the other hand, the amorphous 0.548 obtained by solvent distillation of the previous crystallization filtrate and washing liquid was purified by column chromatography (
Silica gel, chloroform: methanol: vinegar 1 t (95
:5:1)) As a white solid, (2R,3
S)-3(9-fluorenylmethyloxycarbonyl)
7 minnow 2-hydroxypentane 110.3079 was obtained.

Example 19 Boc-L-Va 1-L-Va l -L-3-amino-2-hydroxypentanoyl-0-1eu-L-V
Synthesis of al resin Fmoc-L-valyl-p-alkoxybenzyl alcohol resin 0.59 (contains 0,345-ol of valine) (purchased from Watanabe Chemical) was mixed with dimethylformamide (3
6dXi minutes).

Then 20% piperidine/DMF6a! The mixture was added and stirred for 3 minutes, and after the liquid was discarded, the piperidine treatment was repeated twice (2 times with stirring for 3 minutes and the third time with stirring for 20 minutes). After waste liquid, DM
Washed with F (3×6se×1 min).

Then washed with N-methylpyrrolidone (3 x 6 aeXI minutes). Next item, Fmoc-D-Leu366■ (3 parts)
), HoBt 140■ (3 of our company), and 0.265 ml of diisopropylcarbodiimide (3 of our company) were dissolved in 6 ml of N-methylpyrrolidone and stirred for 31M. After discarding the solution, it was washed with N-methylpyrrolidone (3×6d×1 min), and after discarding the solution, it was washed with DMF (3×6d×I min).

Piperidine deprotection as described above was then performed to obtain Fmo
c-3-amino-2-hydroxy-pentanoic acid 3681
9. HOBt 140cm, diisopropylcarbodiimide 0.265m (3 parts I) was dissolved in 6d N-methylpyrrolidone, and the mixture was stirred for 3 hours. Wash as below, piperidine deprotection, and Fmoc-L-Val in the same manner after washing.
(351j19), Boc-L-Val (225■
) was condensed.

Example 20 Boc-L-Va 1-L-Va I-L-3-amino-2-hydroxypentanoyl-D-1eu-L-V
Oxidation of al resin After washing the above resin with DMF (3X6dXI minutes) DMS
Wash with O (3 x 6 d x 1) L, then add 125 q of pyridine-trifluoro Oi yate and 0.45 ae of acetic anhydride for 6 liters.
- was dissolved in DMSO, added, and stirred for 188 hours. After waste liquid, add DMSO (3x6a+exI minutes), DMF (3x6
After sequentially washing with IIIX 1 minute) and methanol (3X6s!XI minutes), it was dried under vacuum for 2 hours.

Example 21 Synthesis of L-Va 1-L-Va 1-L-3-amino-2-oxopentanoyl D-Leu-L-Va I The dried resin of Example 20 above was mixed with TFA: Phenol 95
: 5 (3x6dx60 minutes). This place 1
The resulting solution was evaporated in vacuo and the residue was dissolved in 3 m methanol and poured onto a column of Sephadex LH-20 (300++d) swollen with methanol. The reaction mixture was developed with methanol, and the first eluting fraction showing UV absorption was collected and evaporated in vacuo.

One tablespoon of the obtained residue was mixed with n-butanol, acetic acid, and water 10
0:1:100 mixture of upper layer 1111jIe and lower layer 1-
Chromatography was performed using a free-flow chromatography device manufactured by Miki Engineering Co., Ltd. using F as a mobile phase. The fractions exhibiting UV absorption were collected and evaporated in vacuum to obtain a colorless powder of the title compound. It was done.

Example 22 Fmoc- of 0.59 as in Example 19.20.21
Using L-valyl resin as a starting material, 3 times equivalent of FmoC-
By sequentially combining amino acids, followed by oxidation and deprotection, L-valyl-prolyl-3-amino-2-oxopentanoyl-〇-〇isyl-1--valine 124.6
I got q.

The physical property values are shown in Table 1.

Example 23 23.6 μl of monovalyl L-buOlyl-3-amino-2-oxopentanoyl-D-leucyl-shi-valine obtained in Example 22 was dissolved in 2d of methanol to obtain 4-phenylbutanoic acid. A 1-methylene chloride solution of anhydride 3011 and 0.050 d of triethylamine were added, and the mixture was stirred at room temperature for 1 hour.

The reaction solution was acidified with acetic acid and purified using a column of Sephadex [H2O (300 ad) filled with methanol.
3-Amino-2-oxopentanoyl-
D-leucyl-d-valine was obtained.

Claims (1)

[Claims] A 3-amino-2-hydroxy fatty acid having the following partial structural formula ▲ Numerical formula, chemical formula, table, etc. ▼ [I] (In the formula, R_1 represents a saturated or unsaturated hydrocarbon group.) The derivative of is oxidized to obtain a 3-amino-2-oxo fatty acid derivative having the following partial structural formula ▲ Numerical formula, chemical formula, table, etc. ▼ [II] (in the formula, R_1 is the same as above). 3, characterized in that the protecting group of the group is removed.
-Production method of amino-2-oxo fatty acid derivative.