JPS6050780B2 - 3-Amino-2-hydroxypropionic acid derivative - Google Patents

Description

Detailed Description of the Invention The present invention is based on the following general formula R1-CH-CH-COOH iI(I) (wherein R is a lower alkyl group having two carbon atoms, a benzyl group, or a halogen atom as a substituent, Indicates a benzyl group having a lower alkyl group or a hydroxy group.

Further, the amino group may be protected with a urethane type protecting group to become a urethane group. ) 3-amino-2
-Hydroxypropionic acid derivative, this compound is bestatin (Bestatin), which is useful as an anticancer agent, etc.
atin) [(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl(S)-leucine] and its analogues.
In the present invention, the lower alkyl group of R is a methyl group,
Examples of the halogen atom include ethyl group, propyl group, butyl group, and chlorine atom, bromine atom, and fluorine atom.

The 3-amino-2-hydroxypropionic acid derivative represented by the general formula (I) of the present invention has the general formula (■) R1-℃H
-CH-CN 1I(■) (In the formula, R is the same as above.

) or a nitrile derivative with its amino group protected can be obtained by hydrolyzing with an acid.

Representative compounds of the nitrile derivatives represented by the above general formula (■) include 3-amino-2-hydroxy-5-methylhexane nitrile 3-amino-2-hydroxy-4
-P-Chlorenylbutyronitrile 3-amino-2-hydroxy-4-P-bromienylbutyronitrile 3-amino-2-hydroxy-4-phenylbutyronitrile 3
-amino-2-hydroxy-4-0-chloroenylbutyronitrile 3-amino-2-hydroxy-4-P-methylenylbutyronitrile 3-amino-2-hydroxy-4-P-butylenylbutyronitrile 3-amino-2-
Hydroxy-4-P-methoxyphenylbutyronitrile 3-amino-2-hydroxy-4-P-butoxyphenylbutyronitrile 3-amino-2-hydroxy-4-(
Examples include 3,4-methylenedioxyphenyl)butyronitrile.

In the case of protecting an amino group, all known amino group protecting groups can be used, but acyl type protecting groups are preferably used, such as formyl, acetyl, trifluoroacetyl, substituted and unsubstituted benzoyl, and urethane type protecting groups. substituted and unsubstituted benzyloxycarbonyl as groups,
Other protective groups such as alkoxycarbonyl and cycloalkanooxycarbonyl having 1 to 6 carbon atoms include substituted and unsubstituted arylsulfonyl, phthalyl, o-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 alkylsulfonic acids such as methanesulfonic acid and ethanesulfonic acid, and arylsulfonic acids such as benzenesulfonic acid and toluenesulfonic acid. It will be done. 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.

For those with protected amino groups that are poorly soluble in aqueous acid solutions, use water-soluble organic solvents such as tetrahydrofuran, dioxane, lower alcohols, acetone, dimethylformamide, dimethylacetamide, etc. to increase solubility. Dimethyl sulfoxide or the like may also be added.

Furthermore, if a protecting group is used and the protecting group decomposes during hydrolysis to produce an electrophile, a cation trapping agent 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, for example, when N-benzyloxycarbonyl-3-amino-2-hydroxy-41P-methoxyphenylbutyronitrile is hydrolyzed in a mixture of equal amounts of concentrated hydrochloric acid and dioxane, 3-amino-2 -Hydroxyphenyl-4-P-hydroxyphenylbutyric acid is obtained.

From the reaction mixture, the desired 3-amino-2- of general formula [1]
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 and neutralize with alkali until the isoelectric point is reached, and if necessary add acetone, methanol, ethanol, etc. Collect the precipitated crystals, or in the case of a non-volatile acid, dilute it to less than 1N with water and pass it through a strongly acidic ion exchange resin to adsorb the desired amino acid. After washing with water, add a volatile solution such as aqueous ammonia. The target substance (1) is eluted with an alkali, concentrated under reduced pressure, and if necessary, acetone, methanol, ethanol, etc. are added to remove the precipitated crystals.
You can get

In the above reaction, when (R)-nitrile derivatives are used as starting materials, amino acids (2RS, 3R
) When a (S)-nitrile derivative is used, a (2RS,3S) body is obtained, and when a (RS)-nitrile derivative is used, a (2RS,3RS) body is obtained.

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

(2RS,3S) or (2RS,3R)-3-amino-2-hydroxy-5-methylhexanoic acid (2RS,3S)
RS)-3-amino-2-hydroxy-4-P-chlorophenylbutyric acid (2RS, 3S) or (2RS, 3R)
-3-amino-2-hydroxy-4-P-bromphenylbutyric acid (2RS,3S) or (2RS,3R)-3-
Amino-2-hydroxy-4-phenylbutyric acid (2RS,
3S) or (2RS,3R)-3-amino-2-hydroxy-4-0-chlorophenylbutyric acid (2RS,3S)
or (2RS,3R)-3-amino-2-hydroxy-4-P-methylphenylbutyric acid (2RS,3S) or (2RS,3R)-3-amino-2-hydroxy-4-
P-butylphenylbutyric acid (2RS, 3S) or (2R
S,3R)-3-amino-2-hydroxy-4-P-nitrophenylbutyric acid (2RS,3S) or (2RS,3
R)-3-amino-2-hydroxy-4-p-hydroxyphenylbutyric acid 3-amino-2-hydroxy-4-(3
, 4-dihydroxyphenyl)butyric acid The above compound can be used as a raw material for bestatin related compounds. For example, benzyloxycarbonyl-(2S,3R)-3-amino-2-hydroxy-4-phenylbutyric acid (hereinafter referred to as "Z" for benzyloxycarbonyl group
, 3-amino-2-hydroxy-4-phenylbutyric acid as “
AHPA”.

) is an optically active form, so in order to obtain this compound, (2RS,3R)-Al(PA is fractionally crystallized and (2S,
3R)-AHPA. So (2RS,
As a result of various studies on fractional crystals of 3R), we first obtained Z-(2RS,3R) obtained by benzyloxycarbonylation of (2RS,3R)-Al(PA) by a conventional method.
It has been found that -AHPA can be fractionally crystallized by forming a salt with brucine in ethyl acetate.

That is, equivalent to Z-(2RS,3R)-AHPA or
A slight excess of brucine is dissolved by heating in ethyl acetate, and if necessary, after filtering, the Oki liquid is allowed to cool or is cooled to precipitate crystals. Optically pure Z-1 (2S
,3R)-AHPA brucine salt is obtained.

If this salt is debrucinated by a conventional method, optically pure Z-(2S,3R)-AHPA can be obtained. Also Z-(
2R,3R)-AHPA and a small amount of Z-(2S,3R)
- Shake the ethyl acetate liquor containing the brucine salt of AHPA with dilute hydrochloric acid to remove brucine, dry the ethyl acetate layer with a dehydrating agent such as anhydrous sodium sulfate, concentrate under reduced pressure, dissolve in a small amount of ethyl acetate, and remove petroleum ether. In addition, optically pure Z-(2R,3R)-
AHPA is obtained. As a result of further research by the present inventors, the benzyloxycarbonylated (2R,3R) derivative of the compound of general formula (1) is more soluble in organic solvents than the (2S,3R) derivative. I discovered a small thing. That is, Z-(2RS,3R)-AFIPA is dissolved in a first soluble solvent at room temperature or warmed, and a second insoluble solvent is added to precipitate crystals to collect the crystals, and this operation is performed as necessary. By repeating , optically pure Z-(
2R,3R)-AlIPA is obtained.

Examples of the first soluble solvent include lower alcohols such as methanol, ethanol, and propanol, ethers such as diethyl ether, diisoproblether, tetrahydrofuran, and dioxane, esters such as methyl acetate, and ethyl acetate, acetone, and methyl ethyl ketone. Ketones such as methylene chloride and chloroform, 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 benzine, 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, the optically impure Z-(2S,3R)-AHPA in the stream liquid is formed into a salt with brucine in ethyl acetate, which is fractionated and crystallized, resulting in optically pure Z-(2S,3R)-AHPA. It can be done.

Furthermore, Z-AHPA used above can be synthesized as follows. For example, in the presence of alkali (2RS, 3R) -
Benzyloxycarbonylation reagent by reaction of AlIPA with benzyloxycarbonyl chloride or in the presence of an organic tertiary base such as triethylamine or N-methylmorpholine in a mixed solvent of dioxane, tetrahydrofuran, acetonitrile, dimethylformamide, etc. and water. For example, benzyloxycarbonyl p-nitrophenyl ester, benzyloxycarbonyl azide, benzyloxycarbonyl N-hydroxysuccinimide ester,
It is obtained by reaction with benzyl S-4,6-dimethylpyrimido-2-ylthiol carbonate and the like.

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

Next, 3 expressed by the general formula (1) obtained as above
- In order to obtain bestatin and its analogues by condensing amino-2-hydroxypropionic acid or its optically active substance with leucine, the condensation is carried out by a known condensation method for peptide bond formation, and then functional groups not involved in the reaction are removed. If it has been protected, the protecting group can be removed. The condensation method of the compound acid represented by the general formula (1) with, for example, leucine includes dicyclohexylcarbodiimide, a carbodiimide method using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, etc., an azide method from hydrazide, ethyl chlorocarbonate, Mixed acid anhydride method using isobutyl chlorocarbonate etc., activated ester method using cyanomethyl ester, vinyl ester, substituted and unsubstituted phenyl ester, thiophenyl ester, hydroxysuccinimide ester etc., O- using acetoxime, cyclohexanone oxime etc. Examples include an acyl hydroxylamine derivative method and an N-acyl compound method using carbodiimidazole.

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, tetrahydrofuran, and dioxane; esters such as ethyl acetate; ketones such as acetone and methyl ethyl ketone; halogenated hydrocarbons such as methylene chloride and chloroform;
Amides such as dimethylformamide and dimethylacetamide, and nitriles such as acetonitrile can be used.
When condensing a compound of general formula (1) 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 that is miscible with water, such as sodium hydrogen carbonate, magnesium oxide, etc. The condensation reaction is preferably carried out in the presence of an inorganic base or an organic tertiary base such as triethylamine or N-methylmorpholine. The protected target products thus obtained are subjected to palladium-catalyzed catalytic reduction, hydrogen bromide in acetic acid, trifluoroacetic acid, hydrogen chloride in organic solvents, saponification with alkali, hydrazine, liquid ammonia, liquid hydrogen fluoride. The protective group can be removed by a known protective group removal method such as, etc., to lead to bestatin and its analogues.

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

The first method is to prepare amides obtained from an α-ami7 acid with a protected amino group and a secondary amine into a metal hydride in an ether such as diethyl ether, tetrahydrofuran, or dioxane at a temperature below 0°C.
) to form an 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. , i.e. 3
-Amino-2-hydroxynitrile.

As the secondary amine used above, N,N-dimethylamine, aziridine, N-methylaniline, imidazoles, pyrazoles, etc. are preferable. Preferred examples of the metal hydride include lithium aluminum hydride, lithium di- or trialkoxyaluminum hydride, and sodium-bis(2-methoxyethoxy)aluminum hydride.

As a protecting group for an amino group, all known protecting groups including an acetyl group and an acyl group including a benzoyl group can be used, but a urethane type protecting group, especially a benzyloxycarbonyl group, is preferable. The second method involves converting the lower alkyl ester of an α-amino acid protected with the same amino-protecting group as above into diethyl ether at a low temperature of -40°C or lower.
Amino groups were protected by reduction with sodium aluminum hydride, diisobutylaluminum hydride, sodium bis(2-methoxyethoxy)aluminum hydride, etc. in ethers such as tetrahydrofuran, hydrocarbons such as toluenehexane, and pyridine. The aminoaldehyde is converted into 3-amino-2-hydroxynitriles in the same manner as above.
Second, the physiological activities of bestatin and its related compounds obtained using the compound of the present invention as raw materials will be described. (4) Test method for inhibitory activity against aminopeptidase B: 1. Measurement of aminopeptidase B activity was performed as described by Hops et al. [V. K. HOPSU
, K. KOMAKINEN, G. G. GLENNER,
ArchivesOfBiOchemistryand
B10phySicsTi, 557 (1966)] was modified.

That is, 1.0 ml of 0.1 M Tris-HCl buffer was added to 0.3 ml of 0.1 mM arginine-1β-naphthylamide.
A mixed solution (PH
7. O) was heated at 37°C for 3 minutes, and then purified using an enzyme purification method according to the method of Hops et al.
Add 0.2 ml of the aminopeptidase B solution purified in , react for 30 minutes at 3rC, and add garnet G°C (0-aminoazotoluene diazonium salt) at a concentration of 1.0m9/ml to a concentration of 1.0%. After adding 0.6 mt of 1.0 M acetate buffer (PH4.2) containing Fin(920) and leaving it at room temperature, the absorbance (a) at 530 m was measured.At the same time, only the buffer containing no analyte was added. The absorbance (b) was measured in a blind manner using the following method, and the aminopeptidase B inhibition rate was calculated by (ba)/b×100.

Results: The inhibition rate at several concentrations of each specimen was determined by the above test method, and 50% inhibition (n) 50) was derived from it.

The results are shown in Table 1. Leu: leucine or its residue AOC: 2-aminooctanoic acid or its residue AIa
: Alanine or its residue Me-1se: β-methyl isoserine (3-amino 4f-2-hydroxybutyric acid) Is
O-Bu-1se: β-isobutyl isoserine (3-amino-2-hydroxy-5-methyl to xanoic acid or its residue Ph-1se: β-phenylisoserine (3
-Amino-2-hydroxy-3-phenylpropionic acid or its residue AHPA (p-C1): 3-amino-2-hydroxy-4-p-chlorophenylbutyric acid or its residue AHPA (0-C1): 3-amino-2- Hydroxy-4-0-chlorophenylbutyric acid or its residue AI (PA(p-Me): 3-amino-2-hydroxy-4-p-methylphenylbutyric acid or its residue AH
PA (p-NO2): 3-amino-2-hydroxy 4-p-nitrophenylbutyric acid or its residue AHPA
(p-0H): 3-amino-2-hydroxy 4-p
-Hydroxyphenylbutyric acid or its residue
Note that the term "residue" refers to the amino acid mentioned above, with one hydrogen atom removed from the amino group, a hydroxyl group removed from the carboxyl group, or one hydrogen atom from the amino group and a hydroxyl group from the carboxyl group. d is shown after removing .

3) Test method for enhancing the transfer of bleomycin to organs by bestatin: 3 male ICR mice (4 weeks old) received 100 mg/K9 subcutaneously of bleomycin B2 alone, and the other 3 mice received 100 m of bleomycin B21OOm9/Kg and bestatin.
g/K9 was administered subcutaneously at the same time, each mouse was sacrificed 1 hour later, and the amount of bleomycin in the serum and organs was determined by Bacillus subtilis (B3cllus subtilis).
) Quantification was carried out by the thin disk method using PCI2l9 as the test bacterium.

result: The above results are shown in Table 2.

Note that the numbers are average values (μ/m1). (C) Bestatin contraceptive effect testing method: 10 mg/K9 of bestatin was injected subcutaneously into ICR-JCL female mice in the estrus stage, and the mice were allowed to cohabit with male mice of the same strain.
For those mice in which mating was confirmed, the female mice were sacrificed two days after vaginal plug formation, and eggs were collected from the oviduct to examine the presence or absence of fertilization.

The fertilized egg at this stage is in the 8 to stent cell stage. result: The above test results are shown in Table 3.

The present invention will be specifically explained below with reference to Examples, 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 were used in the examples.

HOBt: 1-hydroxybenzotriazole DCCD
:N,N''-dicyclohexycarbodiimide HOSU
:N-hydroxysuccinimide 0Bz1●TOsO
H. Benzyl ester p-toluene sulfonate-0
SU: N-hydroxysuccinimide ester 0M
e: Methyl ester-0Nb: p-nitrobenzyl ester BOc: t-butoxycarbonyl DCHA: dicyclohexylamine Note that the Rf values in the examples are expressed using silica gel GF254 plate (manufactured by Merck & Co., Ltd.), n-butanol diacetic acid: water (
4:1:1).

Example 1 (1) Synthesis of (2RS,3R)-AHPA Z-(2R
S,3R)-3-amino-2-hydroxy-4-phenylbutyronitrile (35.4y) was dissolved in concentrated hydrochloric acid (300ml).
11 Dissolved in 300 ml of dioxane, anisole 21.
Add 2y and heat to reflux for 1 full hour.

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

Next, 200 mL of water was added to the residue to remove insoluble matter, and the same amount of acetone was added, followed by adjusting the pH to 5.5 with aqueous ammonia. The mixture was left in a refrigerator overnight, and the precipitated crystals were harvested to obtain (2RS, 3R)-AFIPAl3.66q.

[α] Finished. +16.8F (c=1.25, 1NHC1
), RfO. 23, O. 26 elemental analysis values (ClOHl3
As NO3) Actual value C: 61.06%, H: 6.55
%, N: 6.80% Theoretical value C: 61.52%, H: 6.
71%, N: 7.18% (2) Z-(2S,3R)-A
Synthesis of FIPA (2RS,3R)-AHPAl3.6
Dissolve 6y in 70ml of 1N aqueous sodium hydroxide solution,
While stirring vigorously under ice-cooling, add 70% of benzoyloxycarbonyl chloride 15TILt11N aqueous sodium hydroxide solution.
Divide m1 into 3 portions and add for 3 minutes.

After the dropwise addition, the mixture was vigorously stirred for 1 hour under ice cooling and for 3 hours at room temperature. After the reaction? Add hydrochloric acid to adjust to PHL.

When an oily substance precipitates, it is extracted twice with 100 ml of ethyl acetate, and the ethyl acetate layers are combined, washed with water, and dehydrated and dried over anhydrous magnesium sulfate. The magnesium sulfate was removed, concentrated under reduced pressure, and the residue was recrystallized from ethyl acetate-petroleum ether to give optically impure Z-(2R,3R)-AHPA8.48y.
get.

The mother liquor was concentrated under reduced pressure and crystallized with petroleum ether to give optically impure Z-(2S,3R)-AFIPA8. Obtain O7y. M. p. l42-143℃, [α]? 98+70.2
y (c=1.44, AcOH) The optically impure Z-(2S,3R)-AHPA8. OOy and 10.35y of brucine dihydrate are heated and dissolved in 300ml of ethyl acetate and 10ml of methanol, and the mixture is allowed to stay overnight.

When the liquid is released, the brucine salt of Z-(2S,3R)-AHPA precipitates, which is collected and recrystallized twice from ethyl acetate to obtain pure Z-(2S,3R)-AHPA.
13.51y of brucine salt is obtained. M. p. l443C
l [α] Return. +33.8H (c=1.12, AcOH)
13.5y of this crystal was suspended in 150ml of ethyl acetate,
Brucine was removed with 30 ml of 1N hydrochloric acid, and the ethyl acetate layer was washed with water and dehydrated and dried over anhydrous magnesium sulfate.

Magnesium sulfate was removed, ethyl acetate was distilled off under reduced pressure, and Z-(2S) was recrystallized from ethyl acetate-petroleum ether.
, 3R) - AHPA6.2ly is obtained.

M. p. l54.5℃, [α]? 8+83.5AcO
H) Example 2 Synthesis of (2RS,3R)-AHPA (2RS,3R)-3-amino-2-hydroxy-4-
11.0 da of phenylbutyronitrile hydrochloride was heated under reflux with 200 ml of 6N hydrochloric acid 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 100ml of water to the obtained solid and remove the insoluble matter. Remove, add the same amount of acetone, and remove P with aqueous ammonia.
Adjust to H5.5 and leave in the refrigerator overnight. The precipitated crystals? Example 3 (1) Synthesis of Z-(2S,3S)-AI(PA) is obtained by washing with a water-acetone (11) solution. Z1 (2R
S,? )-AHPA 7.75y and dehydroabiethylamine 6.69y are prepared, and the dehydroabiethylamine salt of Z-(2R,3S)-AFIPA, which is sparingly soluble in ether, is separated.

Concentrate the lemon juice under reduced pressure, and add ethyl acetate to the resulting oil.
Crude Z-(2S,3S)-AHPA by treatment with N-hydrochloric acid
Obtain the crystals.

The crystals were recrystallized twice from ethyl acetate and petroleum ether to obtain pure Z-(2S,3S)-AHPA2. Obtain l6q. M. p. l75~176-C1 [α] Mamoru 98+5.6
. (c=1.39, AcOH) Example 4 Synthesis of (2RS,3R)-Me-1se Z-(2RS,3R)-3-amino-2-hydroxyprobionitrile 9.0y was mixed with Example 1 (1). The same treatment was performed to obtain (2RS,3R)-Me-1se3.16V.

[α]? 8-0.6, (c=1.10,1NHC1),
RfO elemental analysis value (as C4H9NO3) Actual value C
: 40.53%, H: 7.51%, N: 11.95% Theoretical value C: 40.33%, H: 7.62%, N: 11.7
6% Example 5 (2RS, 3R)-1s0-Bu-1se
Synthesis of Z-(2RS,3R)-3-amino-2-hydroxy-5-methylhexanonitrile 2.76y was dissolved in a mixture of concentrated hydrochloric acid 30TrLt and dioxane 30ml,
Add 2.16y 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,
The aqueous layer is then concentrated to dryness under reduced pressure. Dissolve the residue in water,
It is passed through a column packed with 50 ml of Dowex 850x4 (H type) resin of 50 to 100 mesh, and after washing the column with water, it is eluted with aqueous ammonia, and the eluate is concentrated to dryness under reduced pressure. Water-acetone (1:1) was added to the residue, and the precipitated crystals were taken off to give (2RS,3R)-1s0
-Bu-1se0.82y is obtained.

[α]? 8-8.3 When this (2RS,3R)-1s0-Bu-1se is benzyloxycarbonylated, an oily Z-(2RS,3R
)-1s0-Bu-1se is obtained.

Example 6 Synthesis of (2RS,3R)-Ph-1se 3.0y of Z-(2RS,3R)-3-amino-2-hydroxy-3-phenylprobionitrile was added to Example 1
) to (2RS,3R)-Ph-IseO
．． Get 8y.

[α] No. +9.5.

(c=1.0,1NHC1), RfO. l3, O. l7
(Threo and erythro bodies separate on the thin plate.

) Elemental analysis value (as C9H, NO3) Actual measurement value C:
59.18%, H: 6.15%, N: 7.67% Theoretical value C: 59.66%, H: 6.12%, N: 7.73% Example 7 (2RS, 3RS)-AHPA Synthesis of (p-Cl) Z-(2RS,3RS)-3-amino-2-hydroxy-4-p-chlorophenylbutyronitrile 10.3y
is processed in the same manner as in Example 1 (1) (2RS, 3RS)
-AHPA (p-C1) 3.10y is obtained.

RfO. 25 element analysis values (as ClOHl2NO3Cl) Actual values C: 52%, H: 5.17%, N: 5.
99% theoretical value C: 52.29%, H: 5.27%, N:
6.10% of this (2RS,3RS)-Al(PA(p-
When C1) is benzyloxycarbonylated, Z-(2R
S,3RS)-AHpA(p-Cl)(M.p.l4r
c) is obtained. Example 8 (2RS, 3RS)-AHP
Synthesis of A(0-C1) 10.3y of Z-(2RS,3RS)-3-amino-2-hydroxy-4-0-chlorophenylbutyronitrile was treated in the same manner as in Example 1(1) to synthesize (2RS ,3RS)-AHPA(0-C1)4.82
Get V.

RfO. 25. O. 29 (Threo body and erythro body are separated on the thin plate.) Elemental analysis value (ClOFIl2NO
(As 3Cl) Actual value C: 52.01%, H: 5.17
%, N: 5.91% Theoretical value C: 52.29%, H: 5.
27%, N: 6.10% this (2RS, 3RS)-AH
When PA(0-C1) is benzyloxycarbonylated, Z-(2RS,3RS)-AHPA(0-Cl)(M
．． p. 136.5°C) is obtained.

Example 9 Synthesis of (2RS,3RS)-AFIPA(p-Me) Z
-(2RS,3RS)-11.20y of 3-amino-2-hydroxy-4-p-methylphenylbutyronitrile was treated in the same manner as in Example 1(1) to (2RS,3RS)-
AllPA(p-Me) 3.41y is obtained.

RfO. 2O elemental analysis value (as CllHl5NO3) Actual value C: 62.94%, H: 7.01%, N: 6.
79% theoretical value C: 63.14%, H: 7.23%, N:
6.99% this (2RS,3RS)-AHPA(p-M
When e) is benzyloxycarbonylated, Z-(2RS
, 3RS)-AHPA(p-Me) DCHA salt (M.
p. 1666.5-168MC) are obtained. Example 1
Synthesis of 0(2RS,3RS)-AllPA(p-0H) Z-(2RS,3RS)-3-amino-2-hydroxy-4-p-methoxyphenylbutyronitrile 22.0y
is processed in the same manner as in Example 1 (1) to obtain (2RS, 3RS
)-AlIPA (p-0H) 6.73V is obtained.

When this (2RS,3RS)-AHPA(p-0H) is benzyloxycarbonylated, Z-(2RS,3RS
)-AHPA (p-0Z) (M.p.l 38-140℃
) is obtained. Reference Example 1 Synthesis of benzyloxycarbonyl-(2RS,3R)-3-amino-2-hydroxy-4-phenylbutyronitrile. Oily Z-(R)-phenylalani was added to a solution of 20.8y of sodium bisulfite and 50ml of water. Naar approx. 52.
7y is added and the separated material is collected and washed with water and ether to yield the crude sodium salt of 2-benzyloxycarbonylamino-1-hydroxy-3-phenylpropanesulfonic acid 77y.

Suspend 68y of this substance in 250wL1 of water,
After adding ether 50070.1, a solution of potassium cyanide 13y dissolved in 100 ml of water was added 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.

The magnesium sulfate was removed, and the ether was distilled off under reduced pressure to obtain oily benzyloxycarbonyl (2RS, 3R).
-3-Amino-2-hydroxy-4-phenylbutyronitrile 49y is obtained.

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

Reference example 2 (1) Eu- on Z-(2S,3R)-AHPA-(S)
Synthesis of 0Bz1 Z-(2S,3R)-AHPA2.25
y. ．． 945mg of HOBt in 70T of tetrahydrofuran
Dissolve in rLt, (S)-Leu-0Bz1●TOSO
After adding H2.75 da, triethylamine 0.98
Neutralize with mt.

After cooling the solution to -5°C, DCCDl. Add 4 Oy and react overnight. Tetrahydrofuran was distilled off under reduced pressure, 200 ml of ethyl acetate was added, and insoluble materials were filtered off.
After washing in the order of 1N sulfuric acid, water, 5% aqueous sodium bicarbonate solution, and water, it was dehydrated and dried over anhydrous magnesium sulfate.

The magnesium sulfate is removed, the liquid is concentrated under reduced pressure, and the resulting residue is solidified with ethyl acetate-petroleum ether. The solidified product was recrystallized from the same solvent to obtain Z-(2S,3R)-AHP.
Crystal 3.26y of A-(S)-reu-0Bz1 is obtained.
M. p. l22~3℃, [α]? 8+14.22 (c=
1. AcOH) Elemental analysis value (as C3lH36N2O6) Actual value C: 69.66%, H: 7.01%, N:
5.13% theoretical value C: 69.90%, H: 6.81%,
N: 5.26%)) (2S,3R)-3-amino-2
-Synthesis of hydroxy-4-phenylptanoyl(S)-leucine Z-(2S,3R)-AHPA-(S)-?
Dissolve u-0Bz13.22f in 100 mt of methanol, add 100 m9 of palladium black, and perform catalytic reduction at normal pressure for 3 hours.

After removing the catalyst and concentrating the solvent under reduced pressure, the residue was recrystallized from methanol-ethyl acetate to obtain 889 mg of (2S,3R)-3-amino-2-hydroxy-4-phenylptanoyl-(S)-leucine. M. p. 233-236) C (decomposition) [α]? 8-23.56 (c=0.48, Ac0H),
RfO. 48 element analysis values (as Cl6H24N2O4) Actual values C: 62.61%, H: 7.86%, N: 8
．． 83% theoretical value C: 62.31%, H: 7.85%, N
:9.09% Example 31) Z-(2RS,3R)-Me
-1se-(S) Synthesis of Eu-0Bz1 (2RS, 3
R)-Me-1se was benzyloxycarbonylated to form an oily Z-(2RS,3R)-Me-1se1.1
0q and (S) Eu-0Bz1●TOSOHl. 97f
l. ．． HOBt67.5m9, triethylamine 0.7
mt. , DCCD948m9 was treated in the same manner as in Reference Example 2(1) to obtain oily Z-(2RS,3R)-Me-1se-
(S)-re u-0Bz1 is obtained.

This was proven to be unique by thin layer chromatography.

2) Synthesis of (2RS,3R)-3-amino-2-hydroxypropionyl-(S)-leucine Z-(2RS,3R)-Me-1se-(S) obtained in (1) Eu
-0Bz1 was subjected to hydrogenation treatment simultaneously with Reference Example 2 (2) to obtain 765 mg of (2RS,3R)-3-amino-2-hydroxypropionyl(S)-leucine.

[α]? 8-28.7 Elemental analysis value (as ClOH2ON2O4) Actual value C
: 51.49%, H: 8.42%, N: 11.96% Theoretical value C: 51.70%, H: 8.58%, N: 12.0
6% Reference Example 4 (1) Synthesis of (2RS,3R)-3-amino-2-hydroxy-5-methylhexanoyl(S)-leucine by benzyloxycarbonylation of (2RS,3R)-1s0-Bu-1se. The resulting oily Z-(2RS,
3R)-1s0-Bu-1se1.32y and (S)-F
u-0Bz1●TOSOHl. 76y, HOBtO. 6
Oyl triethylamine 0.63ml, DCCDO. 9
2y and processed in the same manner as in Reference Example 2(1) to obtain Z-(2
RS,3R)-1s0-Bu-1se-(S)Eu-
A 0Bz1N oil is obtained.

This oily substance was directly subjected to hydrogenation treatment in the same manner as in Reference Example 2 (2), resulting in 0.5% of (2RS,3R)-3-amino-2-hydroxy-5-methylhexanoyl(S)-leucine.
Get 37y.

[α] Ku98-26.6y (c=1.0, Ac0H)
．． RfO. 43, O. 49 (Threo bodies and erythro bodies separate on the thin plate.

) Elemental analysis value (as Cl3H23N2Ol) Actual value C: 57.01%, H: 9.80%, N: 10.14%
Theoretical value C: 56.91%, H: 9.55%, N: 10.
21% Reference Example 5 (1) Synthesis of Eu-0Bz1 on Z-(2RS,3R)-Ph-1se-(S) (2RS,3R)-Ph-1s
Z-(2) obtained by benzyloxycarbonylation of e
RS,3R)-Ph-1seCm. p. 170-171
℃, [α]? 98+18.5■(c=0.47, Ac0
I()] 250mg and (S)-shiuichi0Bz1●TOS
OH394m9, HOBtl3Omg, triethylamine 0.14ml,. Reference example 2 using 165 mg of DCCD
Z-(2RS,3R)-Ph-
1285 mg of Eu-0Bz on Ise-(S) is obtained.

M. p. 93-95℃, [α]? %-84.7 gain (c=
0.49, AcOH))) (2RS,3R)-3-amino-2-hydroxy-3-phenylpropionyl(
S) Synthesis of monoleucine z-(2RS,3R)-Ph-1
Reference Example 2 (
Hydrogenation treatment was performed in the same manner as in 2) to obtain (2RS,3R)-3
-Amino-2-hydroxy-3-phenylpropionyl-(S)-leucine 45m9 is obtained.

[α]? Wow.

-2.4 (c=0.33, Ac0H), RfO. 36
,O. 45 (Threo and erythro bodies separate on the thin plate.

) Elemental analysis value (as Cl5H22N2O4) Actual value C: 60.22%, H: 7.67%, N: 9.46% Theoretical value C: 61.20%, H: 7.53%, N: 9. 52
%do example 6. )Z-(2RS,3RS)-AHPA(p-C1)-
Synthesis of (S)-Fu-0BzI (2RS, 3RS)-A
Z-(2RS,3RS)-AHPA(p-Cl) obtained by benzyloxycarbonylation of HPA(p-Cl)
) (M.p.l47℃)1.81y and (S)-1j2u
-0Bz1◆TOSOH2.37y. , HOBt8lO
mg triethylamine 0.8477111DCCD1.
Z-(
2RS, 3RS)-AH[11)A(p-Cl)-(S
)-re U-0BzI2.50y is obtained.

M. p. l22~124℃, [α]? 8-12.3-(
c=3.18, Ac0H))) (2RS, 3RS)-
Synthesis of 3-amino-2-hydroxy-4-p-chlorophenylptanoyl(S)-leucine Z-(2RS,3RS)-AHPA(p-C1)-(S
) The above Eu-0Bz11.5q was hydrogenated in the same manner as in Reference Example 2 (2) to obtain (2RS,3RS)-3-amino-2
-Hydroxy-4-p-chlorophenylptanoyl (
S) Obtain 650 mg of monoleucine.

[α]? ? 8-11.3i (c=1.33, Ac0H)
, RfO. 42, O. 52 (Threo and erythro bodies separate on the thin plate.

) Elemental analysis value (as Cl6H23N2O4Cl) Actual value C: 55.88%, H: 7.29%, N: 8.11
Theoretical value C: 56.05%, H: 6.76%, N: 8.1
7% Reference Example 7 (1) Synthesis of (2RS,3RS)-3-amino-2-hydroxy-4-p-methylphenylptanoyl(S)-leucine (2RS,3RS)-AlIPA(p-Me) Z-(2RS,3
RS)-AHPA(p-Me) DCHA salt (M.p.
l66.5-168°C) 1.57y and (S) Upper Eu-0BZ1・TOSOHl. 42y. SHOBt486m
g, triethylamine 0.50Tn1. ．． DCCD61
8mg was processed in a conventional manner to obtain oily Z-(2RS,3R
S)-AHPA(p-Me)-(S)Eu-0Bz1
Obtain quantitatively.

This oily substance was treated in the same manner as in Reference Example 2 (2) (2RS
,3RS)-3-amino-2-hydroxy-4-p-methylphenylptanoyl(S)-leucine 497m9
get.

[α]ξwa. -7.2 gain (c=0.96, Ac0H)
, RfO. 58, O. 53 (Threo and erythro bodies separate on a thin plate.

) Elemental analysis value (as Cl7H35N2O4) Actual value C: 63.08%, H: 8.23%, N: 8.49% Theoretical value C: 63.33%, H: 8.13%, N: 8. 69
Example 8) (2RS,3RS)-3-amino-2-hydroxy-4-p-hydroxyphenylptanoyl (
S) Synthesis of one-leucine (2RS, BinS)-AHPA (p
Z obtained by benzyloxycarbonylation of -0H)
-(2RS,3RS)-AlIPA(p-0Z) (M.
p. l38-140℃) 479 mg and (S) Eu-0
Z-(2RS,
3RS)-AHPA(p-0Z)-(S)-ReU-0B
Obtain Z1450m9.

M. p. 98-99℃, [α]A■-14.0 (c=
0.58, AcOH) Obtained Z-(2RS, 3RS)
-AHPA(p-0Z)-(S)Eu-0Bz140
0 mg was treated in the same manner as in Example 2 (2) (2RS, 3
Eu2l9 on RS)-AI(PA(p-0H)-(S)
Get mg. [α]g? 8-8.8) (c=0.90
, AcOH), RfO. 5l, O. 33 (Threo body and erythro body separate on the thin plate.

Claims (1)

[Scope of Claims] 1 General formula ▲ Numerical formula, chemical formula, table, etc. ▼ (The amino group may be protected with a urethane-type protecting group to become a urethane group.)
-Hydroxypropionic acid derivatives.