JPS5823654A - 3-amino-2-hydroxypropionic acid derivative - Google Patents

Abstract

NEW MATERIAL:A compound shown by the formulaI(R1 is lower alkyl, phenyl, benzyl, etc.; amino group may be protected with a protecting group). EXAMPLE:(2RS, 3R)-3-Amino-2-hydroxy-4-phenylbutyric acid. USE:Useful as an intermediate for bestatin[(2S, 3R)-3-amino-2-hydroxy-4-phenylbutanoyl-(S)-leucine]to be used as a carcinostatic agent, etc. and its analog compound. PROCESS:A nitrile derivative (e.g., 3-amino-2-hydroxy-5-methylhexanenitrile, etc.) shown by the formula II or a nitrile derivative with its amino group protected is hydrolyzed with an acid such as hydrochloric acid, etc. in a temperature range between room temperature and the boiling point of the solvent, to give a compound shown by the formulaI.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention represents the following general formula (wherein R1 is a lower alkyl group, phenyl group, benzyl group, or as a substituent): a benzyl group having a rogene atom, a lower alkyl group, or a nitro group. Moreover, the amino group may be protected with a protecting group. ) 3-
This is an amino-2-hydroxypropionic acid derivative, and this compound is useful as an anticancer agent.
(Bestatin) [: (2S, 3R)-3-
Amino-2-hydroxy-4-phenylbutanoyl-(
81-leucine] and its analogues.

The lower alkyl group in the general formula (Il) of the present invention is a methyl group, an ethyl group, and a probyl group.

Examples of the halogen atom include a butyl group and a chlorine atom, a bromine atom and a fluorine atom.

The 3-amino-2-hydroxypropionic acid derivative represented by the general formula (I) of the present invention is a nitrile derivative represented by the general formula I (wherein R0 is the same as above) or a nitrile with its amine group protected. It can be obtained by hydrolyzing the derivative with an acid.

Representative compounds of the nitrile derivatives represented by the above general formula (1) are:

3-amino-2-hydroxy-5-methylhexanenitrile 3-amino-2-hydroxy-3-phenylpropionitrile 3-amino-2-hydroxy-4-p-thalolphenyl phthyro-, 171J/L73- Amino-2-hydroxy-4-p-bromphenylbutyronitrile 3-amino-2-hydroxy-4-phenylbutyronitrile 3-amino-2-hydroxy-4-o-chloro/l, phenylbutyronitrile 3 , -amino-2-human waxy 4-p-methino♂phenyl phthyronitrile 3-amino-2-hydroxy-4-p-butylphenyl phthyronitrile 3-amino-2-hydroxy-4-p-methoxyphenylbutyro Examples include nitrile 3-amino-2-hydroxy-4-p-butoxyphenylbutyronitrile 3-amino-2-hydroxy-4-(3,4-methylenedioxyphenyl)butyronitrile. −
! As a protecting group for protecting an amino group, all known protecting groups for amino groups can be used, but acyl-type protecting groups such as formyl-acetyl, trifluoroacetyl, substituted and unsubstituted benzoyl, etc., and urethane-type protecting groups are preferably used. substituted and unsubstituted benzyloxycarbonyl as type protecting group,
Other protective groups such as alkoxycarbonyl-cycloalkanooxycarbonyl having 1 to 6 carbon atoms include substituted and unsubstituted sulfonyl, phthalyl-0-nitrophenylsulfenyl-trityl, and the like.

The type of acid used for hydrolysis is not particularly limited as long as it is an acid commonly used for hydrolysis of nitriles. For example, inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, etc., and organic acids include methanesulfonic acid.

Examples include alkylsulfonic acids such as ethanesulfonic acid, and arylsulfonic acids such as benzenesulfonic acid-toluenesulfonic acid.

The concentration of the acid is not particularly limited as long as it can be used for normal hydrolysis of nitriles, but a concentration of 1N or more is preferred.

Those with protected amine groups that are poorly soluble in monoacid aqueous solutions may be treated with water-soluble organic solvents such as tetrahydro-dioxane, lower alcohols ML-7, dimethylformamide, etc. to increase solubility. Dimethylacetamide.

Dimethyl sulfoxide 1. If a protecting group is used and the protecting group decomposes during hydrolysis to produce an electrophile, a cation scavenger such as anisole can be added.

Any temperature from room temperature to the boiling point of the solvent can be used for hydrolysis.

In this hydrolysis, N-benzyloxy-5-dicarbonyl-3-amino-2-hydroxy-4-p-
Hydrolysis of methoxyphenylbutyronitrile in, for example, a mixture of equal amounts of hydrochloric acid and dioxane yields 3-amino-2-hydroxyphenyl-4-p-hydroxyphenylbutyric acid.

3-Amino-2- of the desired general formula CI] from the reaction mixture
To isolate the hydroxypropionic acid derivative, conventional methods for isolating amino acids can be applied.

For example, when hydrolyzing with a volatile acid, remove excess acid by concentrating under reduced pressure, add water, neutralize with alkali in a trench that has an isoelectric point, and if necessary add acetone, methanol, or ethanol. In addition, the precipitated crystals can be placed at the bottom of the furnace, or in the case of non-volatile acids, diluted with water to less than 1N, passed through a strongly acidic ion exchange resin to adsorb the desired amino acid, washed with water, and then volatilized with aqueous ammonia, etc. It is eluted with aqueous alkali,
The desired product (Il) can be obtained by concentrating under reduced pressure and adding acetone, methanol or ethanol if necessary to separate the precipitated crystals.

In the above reaction, when a (5)-nitrile derivative is used as a starting material, C2R8,3R is used as an amino acid.
) unitary is obtained when (S)-nitrile derivative is used, (2'fLS, 3S) unitary is obtained, and (2'fLS, 3S) unitary is obtained when (R8,)-nitrile derivative is used, (2R8°3R8) unitary is obtained. It will be done.

The following are examples of representative products of the present invention obtained in this manner.

(2R8,38)'E or (2R8,3R,)-3-amino-2-hydroxy-5-methylhexanoic acid (2Rs
, 38) or (2FLS, 3R)-3-amino-2
-Hydroxy-3-7enylpropionic acid ("2 R, S, 3'R'S ) 73-amino-2
- Hydrocoat-4-p-chlorophenylbutyric acid (2R8,38) and Fi(2R8,3R)-3-amino-2-hydroxy-4-p-bromphenylbutyric acid (2R8,38)-! Take' (2R8, 3J-3-7
Mino-2-hydroxy-4-phenylbutyric acid (2R8,3
S)i! or < 2Rs, 3R)-3-amino-2-hydroxy-4-o-chlorophenylbutyric acid (2R8,38') or (2R8,3R)-3-amino-2-hydroxy-4-p-methylphenyl Butyric acid (2R8,3s)'i or (2R8,3FL,) -3
-amino-2-hydroxy-4-p-butylphenylbutyric acid C2R8,'38) or (2R8,3R,)-3-amino-2-hydroxy-4-p-nitrophenylbutyric acid (2R8,3S')-E or (2R8,3J-3-amino-2-hydroxy-4-p-hydroxyphenylbutyric acid 3-aminol 2-hydroxy-4-(3,4=dihydroxyphenylbutyric acid) The above compound is used as a raw material for bestatin related compounds. One example that can be used is benzyloxycarbonyl-(2S) as a precursor of bestatin.

3R)-3-amino-2-hydroxy-4-phenylbutyric acid (rA'HP) -3-amino-2-hydroxy-4-phenylbutyric acid (rA'HP)
My name is AJ. ) is an optically active substance, so in order to obtain this compound, it is necessary to fractionally crystallize (2R8°3R)-AHP'A to form (2S, 3R,)-AHPA.

Therefore, as a result of various studies on fractional crystals of (2R8, 3R), we first obtained (2R8,, 3R) using a conventional method.
It has been found that Z-(-2R8,'3J-AHPA obtained by benzyloxycarbonylation of -AI(PA) can be fractionally crystallized by forming a salt with brucine in ethyl acetate. That is, Z-(2R8,3R )-AH
An equivalent amount or a slight excess of brucine to PA is dissolved by heating in ethyl acetate, filtered if necessary, and the filtrate is left to cool or cooled to precipitate crystals.

Optically pure Z-(28,3-
The brucine salt of R)-AHPA is obtained.

If this salt is debrucinated by a conventional method, optically pure Z-(28,3R)-AHPA can be obtained.

Z-(2B, 3B)-A) JPA and a small amount of Z-
(2S, 3R') - Ethyl acetate solution F containing brucine salt of AHPA is shaken with dilute hydrochloric acid to remove brucine, the ethyl acetate layer is dried with a dehydrating agent such as anhydrous sodium sulfate, concentrated under reduced pressure, and a small amount of ethyl acetate is removed. Optically pure Z
-(2R,3R)-AHPA is obtained.

As a result of further research, the present inventors found that (2R,3R) of the compound of the general formula (Il) was benzyloxycarynylated.
It has been found that - derivatives have lower solubility in organic solvents than (28,3R)- derivatives.

That is, Z~(2Rs, 3fL)-AHPA is dissolved in a first soluble solvent at room temperature or heated, and a second insoluble solvent is added to precipitate crystals. By repeating the operation, optically pure Z7(2R,3R)-A)JPA is obtained.

Here, the first soluble solvent is methanol.

Lower alcohols such as ethanol and propatool, ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, and dioxane - methyl acetate,
Esters such as ethyl acetate, acetone, ketones such as methyl ethyl ketone, halogenated hydrocarbons such as methyl chloride, chloroformate, amides such as dimethylformamide and dimethylacetamide, and nitriles such as acetonitrile can be used. As the second insoluble solvent, petroleum hydrocarbons such as petroleum ether, petroleum benzene, and ligroin, aromatic hydrocarbons such as benzene and toluene, and alkanes and cycloalkanes such as hexane and cyclohexane can be used.

On the other hand, optically impure Z-(28,3n)lAI(
pH is 1 by forming a salt with brucine in ethyl acetate as described above.
Optically pure Z-(2S, 3R) by fractional crystallization
-AHPA.

Furthermore, Z-AHPA used above can be synthesized as follows. For example, in the presence of alkali (2R8°3R)-
A) Benzyloxycarbonylation test by reaction of IPA with benzyloxycarbonyl chloride or in a mixed solvent of dioxane, tetrahydrofuran, acetonitrile, dimethylformamide, etc. and water in the presence of an organic tertiary base such as triethylamine or N-methylmorpholine. ! For example, it can be obtained by reaction with benzyloxycarbonyl p-nitrophenyl ester, benzyloxycarbonyl azide, benzyloxycarbonyl N-hydroxysuccinimide ester, benzyl S-4,6-dimethylpyrimidol-2-yl-thiol carbonate, and the like.

Although the method for separating optically active forms using the base AHPA as an example has been described, optically active forms of other amino acids can be similarly separated if necessary.

Next, 3 represented by the general formula (I) obtained as above
Pestatin and its analogs can be obtained by condensing -amino-2-hyperoxypropionic acid or its optically active substance with leucine by a known condensation method for forming a peptide bond, and then If the group has been protected, the protecting group may be removed.

Condensation of a compound acid represented by the general formula (long) with, for example, leucine, using dicyclohexylcarbodiimide,
Carbodiimide method using 1-ethyl-3-(3-dimethylaminopropyl)carbo'□diimide, etc., and azide method from hydrazide.

Mixed acid anhydride method using ethyl chlorocarbonate, isobutyl chlorocarbonate, etc.; active ester method using cyanomethyl ester, vinyl ester, substituted and unsubstituted phenyl ester, thiophenyl ester, hydroxysuccinimide ester, etc.

Examples include the 0-acylhydroxylamine derivative method using acetoxime, cyclohexanone oxime, etc., the N-acyl hydroxyl compound method using carbodiimidazole, etc.

Further, as the solvent used for the condensation, any solvent that does not normally participate in amide bonds can be used. For example, ethers such as diethyl ether and tetrahydrofuran-dioxane, esters such as ethyl acetate, ketones such as acetone and methyl ethyl ketone, methylene chloride,
Halogenated hydrocarbons such as chloroform, amides such as dimethylformamide and dimethylacetamide, and nitriles such as acetonitrile can be used.

When condensing a compound of the general formula (Il) with a protected amino group and an amino acid such as leucine with an unprotected carboxyl group by the active ester method, use an organic solvent miscible with water and add sodium hydrogen carbonate.

The condensation reaction is preferably carried out in the presence of an inorganic base such as magnesium oxide or an organic tertiary base such as triethylamine or N-methylmorpholine.

The thus obtained protected target product was subjected to palladium-catalyzed catalytic reduction of hydrogen bromide in acetic acid.

14- Remove the protecting group by a known protecting group removal method such as trifluoroacetic acid, hydrogen chloride in an organic solvent, saponification with an alkali, hydrazine, liquid ammonia, liquid hydrogen fluoride, etc. can lead to.

The nitrile derivative represented by the general formula (2) can be synthesized as follows.

The first method is to prepare amides obtained from an α-amino acid with a protected amino group and a secondary amine with a metal hydride in an ether such as diethyl ether or tetrahydrone dioxane at a temperature below OC. This is reduced to α-aminoaldehyde with a protected amino group, which is then converted into an adduct such as sodium bisulfite, and then this adduct is reacted with cyanide or directly with prussic acid to form cyanohydrin, That is, 3-
This leads to amino-2-hydroxynitrile.

Preferred secondary amines used above include N,N'-dimethylamine, aziridine, N-methylaniline, imidazoles, and pyrazoles.

Examples of metal oxides include lithium aluminum hydride,
Lithium hydride dib or trialkoxyaluminum,
Sodium-bis(2-methoxyethoxy)aluminum hydride and the like are preferred.

As the protecting group for the amine group, all known protecting groups can be used, including acyl groups including acetyl and benzoyl groups, but urethane-type protecting groups, especially benzyloxycarbonyl groups, are preferred.

In the second method, lower alkyl esters of α-amino acids protected with the same amine-protecting group as described above are heated at low temperatures below 140C to ethers such as diethyl ether, tetraheptadurofuran, hydrocarbons such as toluenehexane, etc. The aminoaldehyde with the amine group protected is prepared by reducing with sodium aluminum hydride, diisobutylaluminum hydride, sodium bis(2-methoxyethoxy)aluminum hydride, etc. in pyridine, and then the 3-
This leads to amino-2-hydroxynitriles.

Next, the physiological activities of bestatin and its related compounds obtained using the compound of the present invention as raw materials will be described.

(5) Inhibitory activity test method for aminopeptidase B Measurement of aminopeptidase B activity was performed by Hopps et al.
.

QLENNER, Archives of Bioch
emistry and biophysics 114
．． 557 (1966)) was modified. - i.e. '0.1mM arginine-β-naphthylamide 0.3 ml K'0.1M Tris-HCl buffer 1
．． A mixed solution (pH 7.0) containing 0.0InC sample was heated at 37C for 3 minutes, and then purified using the enzyme purification method of Hopps et al.
Add 0.2 genus of aminopeptidase B solution purified by
After reacting at 37C for 30 minutes, 1. Garnet QB at a concentration of o1n9/ml
1. Containing C (o-aminoazotoluene diazo triple salt) and containing Twin 20 at a concentration of 1.0%.
After adding 0.6 ml of 0M acetate buffer (pH 4,2) and leaving it at room temperature for 15 minutes.

Absorbance (al) was measured at 530 nm.

At the same time, blind absorbance (bl) was measured using only a buffer solution containing no analyte, and the aminopeptidase B inhibition rate (, b
-a)/bx100.

Results: The inhibition rate at several concentrations of each analyte was determined by the above test method, and 50% inhibition (I Dso ) was derived therefrom. The results are shown in Table 1.

18- 1st Table 1 (28,3R) -AHPA-(S)-Leu'
(Bestachi 7) 0.102 (28,3S)-A
HPA-(S)-Leu” 1.250.56 4 (28,3S)-AHPA-Q()-f,eu
O,045(2S,3R)-AHPA-(R8)-
Aoc6 (2R8,3RJ -Me-Ise-(S
)-Leu7 (2R8,3R)-iso-Bu-Is
e-(S)-Leu” 128 (2R8,'3R
)-Ph-Ise-(S)-Leu9 (2R8,3R
8) -AHPA-(p-CI)-(S)-Leu' O
', 0710 (2R8,3R8)-AHPA-(o
-CI)-(S)-Leu O,4811(2R8
,3R8)-AHPA-(p-Me)-(S)-Leu
6.0112 (28,3R)-AHPA-(p-N
O2)-(S)-Leu 0.0113 (28,3
U-AHPA-(S)-Ala O, 2414(
2R8,3R8)-AHPA-(p-OH)-(s)-
Leu 0.1'0AOC: 2-aminooctanoic acid or its residue Ala: Alanine or its residue Me-Ise: β-methyl isoserine (3-amino-2
-Hydroxybutyric acid) iso-Bu-Ise: β-isobutyl isoserine (3-amino-2-hydroxy-5-methylhexanoic acid or its residue Ph-l5e: β-phenylisoserine (3-amino-
=2-hydroxy-3-phenylpropionic acid or its residue A) (PA(p-CI): 3-amino-2-hydroxy-4-p-chlorophenylbutyric acid or its residue A
HPA (o-CI): 3-amino-2-hydroxy-4-〇-chlorophenylbutyric acid or its residue AHP
A (1)-Me ): 3-amino-2-hydroxy-4-p -J f #Phenylbutyric acid is its residue AHPA (p-N02): 3-amino-2'-hydroxy-4- p-nitrophenylbutyric acid or its residue A
) TPA (p-0H): 3-amino-2-hydroxy-4-p-hydroxyphenylbutyric acid or its residue Note that the term "residue" refers to the amine group of the amino acid mentioned above.
This refers to those obtained by removing one hydrogen atom from a carboxyl group, one from which a hydroxyl group is removed from a carboxyl group, or one from which one hydrogen atom from an amino group and one hydroxyl group from a carboxyl group are removed.

(B) Test method for enhancing the transfer of pleomycin to organs by bestatin: ICR, 3 male strain mice (4 weeks old) were given subcutaneous administration of pleomycin B2 alone at 100 μ/ky; the other 3 mice were given pleomycin. B2100■/kg,! -bestatin 10
0 to 100% were administered subcutaneously at the same time.

Each mouse was sacrificed after 1 hour, and the amount of pleomycin B2 in the serum and organs was determined by measuring the amount of pleomycin B2 in the serum and organs.Results: The above results are shown in Table 2. The numbers are average values (87m1
).

Mark *'BLM-Pleomycin B2 **BST-Bestatin (q) Method for testing the contraceptive efficacy of statin: ICR-JCL female medium mice in estrus are subcutaneously injected with bestatin at a dose of 0/ky to male mice of the same strain. Two days after vaginal plug formation, the mice that were confirmed to have mated were sacrificed with acid, and eggs were collected from the oviduct to check for fertilization.The fertilized eggs at this stage had 8 to 16 cells. It is the period.

Result: The above test results are shown in Table 3.

22-Table 3 The present invention will be explained in detail with reference to Examples below, but the method for protecting functional groups, the method for removing them, and the method for forming peptide bonds are not limited to the methods described in Examples. .

In addition to the above, the following abbreviations are used as abbreviations in the examples:
1-hydroxybenzotriazole DCCD: N,
N'-dicyclohexylcarbodiimide HO8u:N-
Hydroxysuccinimide-0Bzl.TosOH: Benzyl ester I)-) Luenesulfonate 108u: N-Hydroxysuccinimide ester-O
Me: Methyl ester 10 Nb: p-nitrobenzyl ester Boc 2t
-Phthoxycarbonyl DCHA dicyclohexylamine Note that the R and f values in the examples were determined using silica gel GF254 plate (manufactured by Merck & Co., Ltd.), n-butanol:acetic acid:water (
4:1:1) and measured.

Example 1 Synthesis of Z-(2R8,,3R)-AHPA
8,3R)-3-Amino-2-hydroxy-4-phenylbutyronitrile (35.4%) was dissolved in concentrated hydrochloric acid (30%).
Dissolved in 0InC dioxane 300M, anisole 21
．． Add 2p and heat under reflux for 12 hours.

Dioxane is distilled off under reduced pressure, the hydrochloric acid solution is washed with nityl, and the aqueous layer is concentrated under reduced pressure to dryness.

Next, 200 ml of water was added to the residue, insoluble matter was removed with F, and the same amount of acetone was added, followed by adjusting the pH to 5.5 with aqueous ammonia.

@l,”,’,, +16.8° (c=1.25.
IN HCI), RfO, 23°0.26 Elemental analysis value (as C1o Hz3NO3) Actual measurement value C
: 61.06%, H: 6.55%, N: 6.80% Theoretical value C: 61.52%, H: 6.71%, N near, 18
%(21Z-(2S, -3J-AHPA synthesis (2R8
, 3ft) -AHPA, 13.6' 6y- was dissolved in 70ml of IN aqueous sodium hydroxide solution, and while stirring vigorously under water cooling, 15ml of benzoyloxycarbonyl chloride was dissolved.
-IN Add 70 ml of aqueous sodium hydroxide solution in 3 portions over 30 minutes. After the dropwise addition, the mixture was vigorously stirred for 1 hour under water cooling and for 3 hours at room temperature.

After the reaction, 6N hydrochloric acid is added to adjust the pH to -1.

-25= An oily substance precipitates, so extract twice with 100 ml of ethyl acetate, combine the ethyl acetate layers, wash with water, and dehydrate and dry over anhydrous magnesium sulfate.

Magnesium sulfate was removed from the furnace, concentrated under reduced pressure, and the residue was recrystallized from ethyl acetate-petroleum ether to obtain optically impure Z-(
2R,3R)-AHPA 8.489- is obtained. The mother liquor was concentrated under reduced pressure and crystallized with petroleum ether to obtain optically impure Z.
-(28,3B)-AHPA8.07ψ is obtained.

m, p, 142-143C-@:8+70.2°(c
, = 1.44-AcOH) The optically impure Z-(28,3R)-AH obtained above
PA 8. OOp and Bu/L'shi/”dihydrate 10.3
Dissolve 5P in 300M ethyl acetate and 10WL methanol by heating and filter. When the P liquid is allowed to cool, Z-
(28,3B,)-AHPA brucine salt precipitates, which is separated and recrystallized twice from ethyl acetate to obtain pure Z-(2S,,3R)-AHPA brucine salt 13
．． 51fI- is obtained.

m+p, 144 C, ((X) 5 几+ 33.8°
(c = 1.12. AcOH) -26 ~ This crystal 13.57 was suspended in ethyl acetate 150 mA,
Remove brucine with 30 ml of IN hydrochloric acid.

The ethyl acetate layer was washed with water, dehydrated and dried over anhydrous magnesium sulfate, the magnesium sulfate was removed, ethyl acetate was distilled off under reduced pressure, and recrystallized from ethyl acetate-petroleum ether to give Z-
(2S,,'3R)-AHPA6.21F! − is obtained.

! 111.1), 154.5 c-Cc+)5::
+83.5° ('c==1.34, Ac0H) Example 2
゜(2R,S,' 3R)-A) Synthesis of T, PA (2R8
, 3R) f3-amino-2-hydroxy-4-phenylbutyronitrile hydrochloride 11i o y to 6N hydrochloric acid 20
0 ml, and heat to reflux for 4 hours.

Hydrochloric acid is distilled off under reduced pressure, 200 ml of water is added, and the mixture is concentrated again under reduced pressure. Add water Loom to the obtained solid, remove insoluble matter, add the same amount of acetone, and adjust the pH with aqueous ammonia.
Adjust to 5.5 and leave in the refrigerator overnight. The precipitated crystals were separated and washed with a solution of ice water and setone (1:1) to obtain (2R8,3R)-AHPA6.19P.

Example 3゜(1) Synthesis of Z-('28.38)-AHPA Z-< obtained in the same manner as in Example 1(1) and (2)
2R8,3S) -A) Make a salt from 7.75y of TPA and 6.699y of dehydroabiethylamine, and from it make Z-(2R,38)-A of sparingly soluble plum in ether.
The dehydroabiethylamine salt of HPA is separated by furnace. The p solution was concentrated under reduced pressure, and the resulting oil was diluted with ethyl acetate-IN.
Treatment with hydrochloric acid yields crude Z-(28,3B)-AHPA crystals. The crystals were mixed with ethyl acetate-petroleum ether.
Pure Z-(28,38)-AHPA by double recrystallization
2.16 f! − is obtained.

m, p, 175~x 76 c-@5:: + 5.
6° (c = 1.39. AcOH) Example 4° (2'R8,,3R) - Synthesis of Me-Ise Z-(2
R8,3R)-3-amino-2-hydroxypropionitrile 9.0? is processed in the same manner as in Example 1 (1) to obtain (2
R8,3R) -Me-Ise 3.16 P is obtained.

@5-0.6° (C21.10. IN HCI)
, RfO elemental analysis value < C4H9NO3) Actual measured value C: 40.53%, N near, 51%, N: 11.9
5% theoretical value C: 40.33%, N near, 62%, Nu
ll, 76% Example 5゜(2R8,3R) -1so-Bu-Ise synthesis Z
-(2R,S, 3[) -276 of -3-amino-2-hydroxy-5-methylhexanonitrile? with concentrated hydrochloric acid 3
Dissolve in a mixture of 0m/( and 30rrLl of dioxane.

Add 2.167 ml of anisole to this and heat under reflux for 4 hours. Dioxane was distilled off under reduced pressure, the residual liquid was washed with ether, and then the aqueous layer was concentrated to dryness under reduced pressure.

The residue was dissolved in water and passed through a column packed with 50 to 100 mSO of Dow X 50X4 (H type) resin 50M. After washing the column with water, it was eluted with 4N aqueous ammonia, and the eluate was concentrated to dryness under reduced pressure. do.

Water-acetone (1:1) was added to the residue, and the precipitated crystals were collected to give (2R8,':3R)-iso-Bu-Is.
eo, obtain 82%.

@5°-8,3° (c= 1.0. Ac0H), R
f 0118 this (2R8,3R)-iso-Bu-I
Penzyl-29-oxycarbonylation of se yields an oily Z-(2R8,3R)-
iso-Bu-Ise is obtained.

Example 6 Synthesis of Z-(2R8,3R)-Ph-Ise
,3R)-3-amino-2-hydroxy-3-phenylpropionitrile3. Oz is treated in the same manner as in Example 1(1) to obtain (2R8,3R)-Ph-Iseo, 81.

@5-%+9.5° (c = 1.0. IN HCI,
), Rf O,13,0,17 (Threo body and erythro body are separated on the thin plate.) Elemental analysis value (C0H,,
As NO3) Actual value C: 59.18%, H: 6.1
5%, N near, 67% theoretical value C: 59.66%, H:
6.12%, Nia, 73% Example 7 Synthesis of (2R8,3R8)-AHPA (p-CI) Z-(2R8,3R8)-3-amino-2-hydroxy-4-1)-chlor Phenylbutyronitrile 10.3t
was processed in the same manner as in Example 1 (1) (2R8°3R8)
-AHPA (p=cI) 3.10 LiP is obtained. Rf
O, 25 elemental analysis value (C, as R12No3C1) 30- Actual value C: 52.11%, H: 5.17%, N: 5.
99% of this (2R8,'3R8)-AHPA (p
-CI) is benzyloxycarbonylated to form Z-(2
R8,3R8)-'AHPA (p-CI) (m-p
-147C) is obtained.

Synthesis Example 8 of (2R8,3R8)-AHPA(o-CI) Example 1 Process in the same way as (1), (2R8°3'R8
) -AHPA (o-CI) -4 no 82? get.

Rf O,25,0,29 (Threo body and Erimo body separate on the door laminate.) Elemental analysis value (CIo) h2NOa (as C1) Actual measurement value C: 52.01%,) T: 5.17% ,N:5,
91% theoretical value C: 52.29%, H: 5.27%, N
:6.10% this (2R8,3R8)-AHPA('o
When -CI) is converted to benzyloxycarboxylic acid/L/bonylation, -
Z-(2R8,3R8)-7HPA (o-CI) (
m, p, 136.5 C) are obtained.

(2R8,3R8)-AHPA (p-Me) synthesis Z
-(2R8,3R8) -3-amino-2-hydroxy-4-p-methylphenylbutyronitrile 1 i, 2
．． Oy-wo Example 1 (1) I'llt, te(2)
R8゜3R8)-AHPA(p,-Me) 3.419
- get le. Rfo, 20 elemental analysis value (C11, Hts
(as NOa) Actual measured value C: 62.94%, H near,
01%, N: 6.79% theoretical value C: 63.14%, H
Near, 23%, N: 6.99% this (2R8, 3R8)
-AHPA(p-Me) is pendyl-dicarbonylated and Z-(2R8,3R8)7AHPA(p-Me)
The DCHA salt (m, p, 166.5-168 C) is obtained.

Example 10 Synthesis of (2R8,3R8)-AHPA(p-OH) Z-
(2R8,3R,S)-3-amino-2-hydroxy'
/ -4-p-methoxyphenylbutyronitrile22.
0p is 1111 as in Example 1 (1), (2R8゜3R8) -AHPA (p-OH), 6.739
− is obtained.

? (2R8,3BS)-AHPA(p-OH)
When subjected to bare zyloxycarbonylation, +z-(2R
s, 3R8)-AHPA(p-0Z) (m, p, 13
8-1' 40 C) is obtained.

Reference Example 1 Synthesis of benzyloxycarbonyl-(2R8,3R)-3-amino-2-hydroxy-4-phenylbutyronitrile Sodium bisulfite 20.8! Z-(2)-phenylalaninal in oil form in a solution of i' and 50 ral of water.
．． Add 7P, divide the separated substance, and add water.

and washing with ether to obtain crude sodium salt 77P of 2-benzyloxycarbonylamino-1-hydroxy-3-phenylpropanesulfonic acid. This substance 68?
was suspended in 250M of water, cooled to 10-12C, and 500 rnl of Emel was added, then potassium cyanide 131 was added to 100mA of water! Add the solution dissolved in water dropwise over 30 minutes.

After the dropwise addition, the solution was allowed to stand at room temperature for 3 hours, and then the ether layer was separated, washed with brine, and dehydrated and dried over anhydrous magnesium sulfate.

33- Magnesium sulfate is removed from the furnace and ether is distilled off under reduced pressure to obtain oily benzyloxycarbonyl-(2R8°3R)-
3-Amino-2-hydroxy-4-phenylbutyronitrile 49FI- is obtained.

Other nitrile derivatives used in the present invention can also be obtained in the same manner as above.

Reference example 2゜(11Z-(28,3R)-AHPA-(81-Leu
Synthesis of -OBzl Z-(28,3R)-AHPA 2.
Dissolve 25 f/-HoBt 945■ in 70 ml of tetrahydrofuran and prepare isl -Leu-OBzl IIT
After adding os OH2,759-, neutralize with 0.98 ml of triethylamine. After cooling the solution to -5C, DCCD 1.4 of total processing was allowed to react overnight. Tetrahydrofuran was distilled off under reduced pressure, 260 ml of ethyl acetate was added, and insoluble materials were removed. After washing in the following order: IN sulfuric acid, water, 5% hydrogen carbonate solution, and then water, dehydrate and dry with anhydrous magnesium sulfate. The magnesium sulfate is removed, the P solution is concentrated under reduced pressure, and the resulting residue is solidified with ethyl acetate-petroleum ether. Solidification-3
4 = Recrystallize the product in the same solvent to form Z-(2S,3R)-A H
Crystal of P A 7 (S) -Leu -OBzl 3.
Get 26p m, p, 122~3 c-Cc,)5”
,: +14.2° (-c=1-AcOH) elemental analysis value (C31H36N206 L'C)'.

Actual value C: 69.66%, H near, 01%, , N
: 5.1.3% theoretical value C: 69.90%, H: 6.
81%, N: 5.26% (2+ (28,3J-3-
Amino-2-hydroxy-4-phenylbutanoyl-(
Synthesis of S)-leucine Z (28,3FL) AH
PA, 48) Dissolve Leu-OBzl 3.221 in 1.00 ml of methanol and add palladium black 100r.
Add n9 and perform catalytic reduction at normal pressure for 3 hours. The catalyst was removed from the furnace, the solvent was concentrated under reduced pressure, and then recrystallized from methanol-ethyl acetate to obtain (2s, 3R)-3-amino-2-hydroxy-4-phenylbutanoyl-(81-leucine 889).

m-p,, 233-236 C (decomposition) @57:
23.5° (c=0.4.8. ACOH), Rf
Q, 48 elemental analysis value (C16Hu N2O4
') Actual measurement value C: 62.61%, H near, 86%, N
: 8.83% theoretical value C: 62.31%, H near, 85
%, N: 9.09% Reference example 3°. '(1) Z-(2R8,3R)-Me-Ise
-Synthesis of (S)-Leu-OBzl.

(2R8,3R) -Me -Ise was converted into an oily Z'-(2R8,3R-
Me-Ise 1.10 fl- and (S) -Leu
-0BzJ ・TosOH1,975', If-JO
Bt 6757Q-triethylamide 70.7~l.

DCCD9481157 was treated in the same manner as in Reference Example 2 (1) to obtain oily Z -(2R8,3R) -Me-Ise.
-(81-Leu -0Bzl is obtained.

This was proven to be unique by thin layer chromatography.

(21(2R8,3R)-3-amino-2-hydroxypionyl-(S)-, Z-(2R8,3R)-Me-Ise-(
81-Leu-013zl as reference example 2 (2)
At the same time, hydrogenation treatment is performed to obtain (2R8,3R)-3-aquinone-2-hydroxyph, lopio, nil-(St-leucine 765).

@5:: -2,8,7°(c==1.09. Ac0
H) Elemental analysis value (as C+o R20N204) Actual value C: 51.49%, H: 8.42%, N
: 11.96% theoretical value C: 51.70%, I
-1: 8.58 and N: 12.06% Reference Example 4゜(1) Synthesis of (2R8,3R)-3-amino-2-hydroxy-5-methylhexanoyl-(S)-leucine (2R8, An oily Z= (2BS) obtained by benzyloxycarbonylation of 3R)-iso-Bu-Ise.

3 R) -1so-Bu-Ise 1.32 F and (
81-Leu -0Bzl aTos OH1,76P
, HOBt O,6'Ofl-, triethylamine 0
．． Reference example 2 using 63mL DCCD 0.92 f
Process in the same manner as (1) to obtain Z −(2R8,3R) −1
An oil of so-Bu7 l5e-(S)-Leu-OBzl is obtained.

This oily substance was subjected to hydrogenation treatment in the same manner as in Reference Example 2 (2). Obtain 75L.

@52I: -26,6° (c=1.0. Ac0H)
, Rf O,43,,,0,49 (Threo body - Eri 1
0 bodies separate on the thin plate.

37- Elemental analysis value (9 actual measured value as C13R23N204
C: 57.01%, )J: 9.80%, N: 10.1
4% theoretical value C: 56.91%, H: 9.55%, N
: 10.21% Reference example 5° (11Z -(2R8,3R) -Ph-Ise-(8
1=Synthesis of Leu -0Bzl Z -(2R8,3J -Ph obtained by benzyloxycarbonylation of (2R8,3B)-Ph-Ise
-Ise [m, p-170~l 71 tll',
@57: +18.5° (c=-0,47,'Ac0H
) ] 25 om9 and (S) -Leu-OBzl
・TosOH394mg-HOBt 130mg-)
IJ: r-f Luami 70.14ml, DCCD
Z-
(2R8,'3R)-Ph-l5e-(81-L
Obtain eu-OBzl 285 mQ.

m-p, 93z95C, @5弗-84.7° (c=0
．． 49. Ac0T-1')(2) (2R8,3R
)-3-Amino-2-hydroxy-3-phenylpropionyl-(S)-leucine synthesis Z -(2R8,3R)-Ph -l5e-(81-L
eu -OBzl) 200Tng was hydrogenated in the same manner as in Reference Example 2 (2) to give 38- (2R8,3J-3-amino-2-hydroxy-3-phenylprohionyl-(S)-leucine 45)
get.

(b) 59 sous 2.4° (c=0.33. Ac0H)
, Rf O,36,0,45 (Threo body and Eri21 body are separated on the thin plate.) Elemental analysis value (C15H2□
As N2O4] Actual value C: 60.22%, H near,
67%, N: 9.46% theoretical value C: 61.20%, H
Near, 53%, N: 9.52% Reference example 6° (11Z-(2R8,3R,5)=AHPA(p-ct
) -(sl -Leu-OBzl Synthesis (2'R8,3R8)-AHPA(p-ct) is synthesized by benzyloxycarbonylation of Z-'(2R8゜3
R8)-AHPA(1)-CI) (m, p, 147
tT )1.81 P and (81-Leu -0Bzl
-TosOH2,37f-HOBt 810η triethylamine O184d, DCCI) 1.03
8) AHPA(p C1) (S)-Leu
-0Bzl Get 2.50g.

ml, p-122-124 c, ■s:W-12,
3° (c=3.18. AcOH) (2) (2R8
,3R8) -3-amino-2-hydroxy-4,-1
)-Chlorphenylbutanoyl-(S)-leucine synthesis Z-(2R8,3R8)-AHPA(p-CI)-(S
i -Leu'-0Bzl 1.5 f in Reference Example 2 (2
) was hydrogenated in the same manner as (2R8,3R8)-3-
650 m9 of amino-2-hydroxy-4-1)-chlorphenylbutanoyl-(81-leucine are obtained.

(b) 5b: -11.3° (C2 1.33. kcOH
), Rf O, 42, 0, 52 (threo, Eli 21
The body separates on the lamina. ) Elemental analysis value (C16H2
3N204 C1) Actual value C: 55.88%,
H near, 29%, N: 8.11c/b theoretical value C: 56
．． 05%, H: 6.76%, N: 8.17% Reference example 7゜(11(2R8,3R,S)-3-amino-2-hydroxy-4-p-methylphenylbutanoyl-(81
- Synthesis of leucine (2R8, 3R8) = AHPA (p -Me ) obtained by benzyloxycarbonylation Z-(2R8
゜3 R8)-AHPA (p-Me) DCHA salt (
m, p-166,5~1681T) 1.57 P and (
81=Leu −0Bzl −TosOH1,42f
-HOBt 486■, triethylamine 0.50mL
DCCD613my was processed using a conventional method to obtain oily Z.
-(2R8,3R8) -AHPA (p-Me)
-(St-Leu-0Bzl was obtained quantitatively.

This oily substance was treated in the same manner as in Reference Example 2 (2) (2R8
,3R8)-3-amino-2-hydroxy-4-p =
Methylphenylbutanoyl (81-leucine 497m
get g.

(b), -7.2° (c=0.96. Ac0H)
, Rf O,58,0,53 (threo body, Eri21 body is separated on the thin plate.) Original elemental analysis value (C17R35
As N20a) Actual measurement value C roller 308%, H: 8.2
3%, N: 8.49% theoretical value C: 63.33%, H:
8.13%, N: 8.69% Reference example 8゜(11(2R8,3R8)-3-amino-2-hydroxy-479-hydroxyphenylbutanoyl-(81-
Synthesis of leucine (2R8,3R8)-Z-(2FL8゜4l) obtained by benzyloxy7carbonylation of AHPA(p-OH)
-3R8)-AHPA(p-0Z) (m, p, 138
~140 U) 479 mg and (S)-Leu'-0B
zl-TosOH472~9. HOBt1621n9
．． Triethylamine 0.168 ml, DCCD20
Z-(
2R8,3R8)7AHPA(p-OZ)-(S)-L
Obtain 450 mg of eu-OBzl.

m-p-98~99C+ @57: ' 140° (c
~0.58. Ac0H) obtained Z-(2R8,3
R8)-AHPA(p-OZ)-(81-Leu-OB
zl 400-9 was treated in the same manner as in Example 2 (2) to obtain (
2R8,,3R8)-AHPA(p-OH)-(S)-
Leu 219-9 is obtained.

@s:: '8.8°(c-0,9,O,Ac0H)
, Rf 0151.0.33 (Threo body and Eri 21 body are separated on the thin plate.) Elemental analysis value (C+e R2
4N2.0s) Actual value C: 59.38%, H near 23%, N: 8.95% Theoretical value C: 59.24%
, I (near, 46%, N: 8.64% 42-

Claims (1)

[Claims] (1) General formula (in the formula R,, tri lower alkyl group to phenyl group, to 7
It represents a syl group 'E or a benzyl group having a halogen atom, lower alkyl group or nitro group as a substituent. Moreover, the amino group may be protected by a protecting group C-protection. ) A 3-amino-2-hydroxypropionic acid derivative represented by: