JPH09136875A - 2-episiastatin b derivative as metastasis inhibitory substance and its production - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new compound as a metastasis inhibitory substance, capable of markedly inhibiting metastasis, thus useful for cancer therapy. SOLUTION: This new compound is expressed by formula I (R<1> is acetyl or trifluoroacetyl; R<2> is amino or guanidino), e.g. (3R,4S,5S,6R)-6ace tamido-4- amino-5-hydroxypiperidine-3-carboxylic acid. The compound of formula I is obtained by the following process: siastatin B of formula II is converted into an N-protected 4-deoxy-3-enesiastatin B by a known method, and the N-protected 4-deoxy-3-enesiastatin is esterified and then reacted with trichloroacetonitrile into a compound of formula III (R<a> is a carboxyl-protecting group) which is then hydrolyzed, and the trichloroacetyl group is eliminated and then the imino- protecting group followed by carboxyl-protecting roup are also eliminated to obtain the objective compound (R<1> is acetyl; R<2> is amino).

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel 3-epiastatin B derivative having a cancer cell metastasis inhibiting activity,
Specifically, (3R, 4S, 5S, 6R) -6-acetamido-
4-amino-5-hydroxypiperidine-3-carboxylic acid,
(3R, 4S, 5S, 6R) -6-acetamido-4-guanidino-5-hydroxypiperidine-3-carboxylic acid, (3R
R, 4S, 5S, 6R) -4-Amino-5-hydroxy-6-
Trifluoroacetamidopyridine-3-carboxylic acid,
(3R, 4S, 5S, 6R) -4-Guanidino-5-hydroxy-6-trifluoroacetamidopiperidine-3-carboxylic acid and pharmaceutically acceptable salts thereof. The invention also relates to methods for their production.

[0002]

2. Description of the Related Art Conventionally known carcinostatic agents mainly consist of drugs having a cytotoxic active substance that kills malignant cells as an active ingredient or an active substance that kills malignant cells through the human immune system as an active ingredient. Met. Many anticancer agents of this type have been studied and used clinically. However, conventional anticancer agents are not yet sufficient for treating cancer.
In addition, for solid cancer, many treatments such as surgery and radiation are also performed. On the other hand, the metastasis of cancer reaches the target tissue through (a) release from the primary focus of cancer cells, (b) dissemination of cancer cells into blood vessels, (c) stagnation in capillaries, embolization, etc. It is known to be a process that consists of cancer cells initiating growth.

[0003]

The effectiveness of various current cancer therapies is expected to be further enhanced by suppressing the metastasis of cancer cells. However, there are few active substances that mainly act to suppress the metastasis of cancer cells, and none of them are clinically used. Therefore, it is strongly desired to provide a new compound having low toxicity, water solubility, and a strong cancer metastasis-inhibiting action, which is effective in treating and preventing cancer.

[0004]

[Means for Solving the Problems] The present inventors have advanced the research for the purpose of obtaining a novel cancer cell metastasis-suppressing substance, and have conducted diligent research focusing on the cystatin B derivative. As a result, they succeeded in synthesizing a novel 3-episistatin B derivative represented by the following general formula (I) having excellent cancer cell metastasis inhibiting activity, and found a method suitable for producing these novel compounds. It was This completes the present invention.

In the first aspect of the present invention, the general formula (I) (Wherein R ¹ is an acetyl group or a trifluoroacetyl group, and R ² is an amino group or a guanidino group), and a 3-epicistatin B derivative and a pharmaceutically acceptable salt thereof are Provided.

The 3-epiciastatin B derivative of the general formula (I) according to the present invention includes the following four compounds.

(1) Compound No. 1: the following formula (3R, 4S, 5S, 6R) -6-acetamide
-4-Amino-5-hydroxypiperidine-3-carboxylic acid [corresponding to a compound of the general formula (I) in which R ¹ is an acetyl group and R ² is an amino group].

(2) Compound No. 2: (3R, 4S, 5S, 6R) -6-acetamide
-4-guanidino-5-hydroxypiperidine-3-carboxylic acid [corresponding to a compound of the general formula (I) in which R ¹ is an acetyl group and R ² is a guanidino group].

(3) Compound No. 3: (3R, 4S, 5S, 6R) -4-amino-5-hydroxy-6-trifluoroacetamidopiperidine-3-
Carboxylic acid [corresponding to a compound of the general formula (I) in which R ¹ is a trifluoroacetyl group and R ² is an amino group].

(4) Compound No. 4: (3R, 4S, 5S, 6R) -4-guanidino-
5-Hydroxy-6-trifluoroacetamidopiperidine-3-carboxylic acid [corresponding to a compound of the general formula (I) in which R ¹ is a trifluoroacetyl group and R ² is a guanidino group].

The salts of the compounds of general formula (I) according to the first aspect of the invention include salts with pharmaceutically acceptable metals at their carboxylic acid groups, such as alkali metal salts, such as sodium salts or alkaline earth salts. There are metal salts such as calcium and ammonium salts. The general formula (I)
Also included is an acid addition salt at the imino group and / or guanidino group of the compound of, especially a pharmaceutically acceptable mineral acid,
For example, there are pharmaceutically acceptable acid addition salts with hydrochloric acid, sulfuric acid, or organic acids such as acetic acid, propionic acid and the like.

Next, the physicochemical properties of the compound according to the first aspect of the present invention (as a hydrochloride salt) will be shown. (A) (3R, 4S, 5S, 6R) -6-acetamido-4
- dihydrochloride salts of amino-5-hydroxy-piperidine-3-carboxylic acid (Compound No.1): Color and shape: colorless amorphous solid Molecular _{Formula: C 8 H 15 N 3 O} 4 · 2HCl Specific rotation: [alpha] ²⁶ _D + 31.8 ° (c 0.19, water) ¹ H-NMR spectrum (400 MHz, D ₂ O, δppm): 2.13 (3
H, s, NHCOC H ₃ ), 3.20 (1H, dt, J = 3.9, 11.5 Hz, 3-H),
3.28 (1H, t, J = 12.0 Hz, 2-Hax), 3.61 (1H, dd, J = 3.9,
12.0 Hz, 2-Heq), 3.98 (1H, dd, J = 2.7, 11.5 Hz, 4-
H), 4.32 (1H, t, J = 2.7 Hz, 5-H), 5.45 (1H, d, J = 2.7 H
z, 6-H) Solubility: Well soluble in water.

(B) (3R, 4S, 5S, 6R) -6-acetamido-4-guanidino-5-hydroxypiperidine-
Dicarboxylic acid salt of 3-carboxylic acid (Compound No. 2): Color and shape: colorless amorphous solid Molecular formula: C ₉ H ₁₇ N ₅ O ₄ .2HCl Specific rotation: [α] ²⁶ _D + 16.8 ° (c 0.26, water) ¹ H-NMR spectrum (400 MHz, D ₂ O, δppm): 2.31 (3
H, s, NHCOC H ₃ ), 3.35 (1H, dt, J = 4.4, 11.0 Hz, 3-H),
3.54 (1H, t, J = 12.0 Hz, 2-Hax), 3.69 (1H, dd, J = 4.4,
12.0 Hz, 2-Heq), 4.38 (1H, t, J = 3.2 Hz, 5-H), 4.45
(1H, dd, J = 3.2, 11.0 Hz, 4-H), 5.52 (1H, d, J = 3.2 H
z, 6-H) Solubility: Well soluble in water.

(C) Dihydrochloride of (3R, 4S, 5S, 6R) -4-amino-5-hydroxy-6-trifluoroacetamidopiperidine-3-carboxylic acid (Compound No. 3): Color and shape: Colorless amorphous solid Molecular formula: C ₈ H ₁₂ F ₃ N ₃ O ₄ .2HCl Specific rotation: [α] ²⁹ _D + 22.1 ° (c 0.36, 1N HCl) ¹ H-NMR spectrum (400 MHz, D ₂ O , δppm): 3.07 (1
H, dd, J = 4.0, 11.2 Hz, 3-H), 3.15 (1H, t, J = 12.7 Hz,
2-Hax), 3.50 (1H, dd, J = 4.0, 12.7Hz, 2-Heq), 3.93 (1
H, dd, J = 2.9, 11.2 Hz, 4-H), 4.25 (1H, t, J = 2.9 Hz,
5-H), 5.41 (1H, d, J = 2.9 Hz, 6-H) Solubility: Well soluble in water.

(D) Dihydrochloride of (3R, 4S, 5S, 6R) -6-trifluoroacetamido-4-guanidino-5-hydroxypiperidine-3-carboxylic acid (Compound No. 4): Color and shape: Colorless amorphous solid Molecular formula: C ₉ H ₁₄ F ₃ N ₅ O ₄ .2HCl Specific rotation: [α] ²⁹ _D + 1.3 ° (c 0.47, 1N HCl) ¹ H-NMR spectrum (400 MHz, D ₂ O , δppm): 3.27 (1
H, dt, J = 4.1, 11.0 Hz, 3-H), 3.38 (1H, t, J = 12.0 Hz,
2-Hax), 3.55 (1H, dd, J = 4.1, 12.0Hz, 2-Heq), 4.25 (1
H, t, J = 3.9 Hz, 5-H), 4.39 (1H, dd, J = 3.9, 11.0 Hz,
4-H), 5.42 (1H, d, J = 3.9 Hz, 6-H) Solubility: Well soluble in water.

The first test example below demonstrates that the compound of the general formula (I) according to the present invention has a cancer cell metastasis inhibiting activity.

Test Example 1 The test for evaluating the cancer cell metastasis inhibitory activity of the compound of formula (I) according to the first aspect of the present invention is the method of Kijima-Suda et al.
("Cancer Research", 46, 858-862, 1986).

The test method is as follows. That is, from the melanoma B16 strain, which is a tumor of mouse,
dler) Method in Cancer Research, Vol. 15, 399
-Page 439, 1978), a highly metastatic melanoma B16 strain was selected and used as a test cancer cell.

First, the B16 high-metastatic strain was planted in Dulbecco's medium supplemented with fetal bovine serum and incubated at 37 ° C. for 24 hours in the presence of 5% CO _2.
And cultured. Then, the compound of the general formula (I) was added as a reagent at each concentration shown in Tables 1 and 2 below,
Cultures were incubated at 37 ° C in the presence of 5% CO ₂ for 72 hours or 72 hours. Then, the grown cells were peeled off from the culture vessel with trypsin and EDTA solution. The cancer cells thus treated and proliferated with the above-mentioned agents were suspended in a balanced salt solution to give a cell suspension containing 1 × 10 ⁶ cells per ml as living cells.

0.1 ml of this cancer cell suspension was administered to the tail vein of a BDF ₁ mouse (female, 7 weeks old) for transplantation. After breeding for 14 days, the mice in the reagent-treated group were subjected to laparotomy to remove the lungs, and the colonies of the B16 high transfer strain formed on the surface and inside the lungs were counted. Furthermore, it compared with the control group which did not perform drug treatment. B16 with lung metastases in its treated and control groups
The number of colonies of the high transfer strain and the metastasis inhibition rate (%) calculated from the number of colonies are shown in Tables 1 and 2 below as test results.

[0021]

[0022]

As is clear from the above test results, (3R, 4S, 5S, 6R) -6-acetamido-4-amino-5-hydroxypiperidine-3-carboxylic acid (compound of formula (I) according to the present invention (compound No. 1) dihydrochloride, (3R, 4S, 5
S, 6R) -6-acetamido-4-guanidino-5-hydroxypiperidine-3-carboxylic acid (Compound No. 2) dihydrochloride, (3R, 4S, 5S, 6R) -4-amino-5-hydroxy- 6-trifluoroacetamidopiperidine-3-carboxylic acid (compound No. 3) dihydrochloride and (3R, 4S, 5
S, 6R) -4-guanidino-5-hydroxy-6-trifluoroacetamidopiperidine-3-carboxylic acid (compound N
o.4) Dihydrochloride reduces the number of cancer cell colonies formed in the lung as the drug concentration for cancer cell treatment by these increases, and strongly suppresses the metastasis of cancer cells to the lung. .

The compound of the general formula (I) having a cancer cell metastasis-inhibiting activity or a salt thereof according to the present invention generally extremely remarkably suppresses the diffusion and metastasis of cancer cells. Therefore, by simultaneously administering the compound of the present invention or a salt thereof at the same time as an existing anti-cancer agent, or by administering it during surgery or radiation therapy, the anti-cancer effect can be remarkably enhanced, which is extremely useful as a method for treating cancer. It is a drug.

Furthermore, a production method for producing a 3-epistatinstatin B derivative of the general formula (I) which is a novel cancer cell metastasis inhibitor will be described.

To produce the 3-epiciastatin B derivative of the present invention, cystatin B is used as a raw material. Cyastatin B is a radiobacterium Streptomyces verticillus Streptomyces verticillus
var. quintum ) ( Mikken Kenkyukai No. 507) is cultured and collected from the culture solution (Japanese Patent Publication No. 55-46714).

The first (3R, 4S, 5S, 6R) -6-acetamido-4-amino-5-hydroxypiperidine-3-carboxylic acid of formula (Ia) according to the present invention (compound No. 1) ) And the formula (Ib) (3R, 4S, 5
S, 6R) -6-acetamido-4-guanidino-5-hydroxypiperidine-3-carboxylic acid (Compound No. 2) can be prepared from Cyastatin B as N-protected-4-deoxy-3-
It can be produced by using Enciastatin B as a starting compound and chemically converting it in several steps.

In the second aspect of the present invention, a method of producing the compound of the present invention represented by the formula (Ia) is represented by the following formula (II) The following formula (III) derived by the known method from cystatin B represented by (Wherein R represents an imino protecting group)
The carboxyl group of -4-deoxy-3-enciastatin B is reacted with an esterifying agent to esterify it, and the compound of the following formula (IV) (Wherein R has the same meaning as described above and R ^a represents a carboxyl-protecting group which is an ester-forming group), an ester compound is produced, and then the compound of formula (IV) is reacted with trichloroacetonitrile. Thereby forming an oxazoline ring, (Wherein R and R ^a have the same meanings as described above), and then the compound of formula (V) is hydrolyzed to open the oxazoline ring, whereby the following formula (VI) (Wherein, R and R ^a have the same meanings as defined above) to produce a compound represented by the further trichloroacetyl group elimination from the compound of formula (VI) the following formula (VII) (Wherein R and R ^a have the same meaning as described above), and the imino protecting group (R) and carboxyl protecting group (R ^a ) are removed from the compound of formula (VII). The following formula (Ia) characterized by separation (3R, 4S, 5S, 6R) -6-acetamide
-4-amino-5-hydroxypiperidine-3-carboxylic acid,
Alternatively, a method for producing the salt is provided.

In the second method of the present invention, 1-N-protected-4-deoxy-3-ene-siastatin B of the general formula (III) used as a starting compound is the cystatin B of the above formula (II).
Nishimura et al. "Natural Product Lett."
1, pp. 39-43 (1992), and Kudo et al .; "J. Antibi.
otics ", Volume 46, pages 300-309 (1993), and JP-A-4-89.
It can be prepared by the method described in Japanese Patent Publication No. 481. For example, 4-
Deoxy-3-en-1-N- (tert-butoxycarbonyl) siastatin B is a di-tertiary butyl carbonate of siastatin B in N, N-dimethylformamide in the presence of an organic amine such as triethylamine. And the like and reacted at 5 to 100 ° C. for 2 to 5 hours.

Next, a method for carrying out each step of the second method of the present invention will be described. The imino protecting group (R) in the compound of the enolized cystatin B of the general formula (III) is preferably an alkoxycarbonyl group, an aryloxycarbonyl group or an aralkyloxycarbonyl group, and particularly preferably a tertiary butoxycarbonyl group. It is convenient. The imino protecting group (R) can be introduced by a conventional method.

First, in order to protect the carboxyl group of the compound of formula (III), the compound of formula (III) is reacted with an esterifying agent which introduces an ester-forming group into the carboxyl group.
Conveniently, the esterifying agent is a compound that can be used to introduce ^a diphenylmethyl group, which is easily eliminated by alkaline hydrolysis, as a carboxyl protecting group (R ^a ), for example, diphenyldiazomethane.

When diphenyldiazomethane is used as the esterifying agent, 1-N-protected-4-deoxy-3-enciastatin B represented by the formula (III) is dissolved in a mixed solvent of dichloromethane and methanol, Diphenyldiazomethane can be added to the solution to carry out the esterification reaction at room temperature, which gives the ester compound of formula (IV). It should be noted that the above-mentioned esterifying agent can generally be a (C ₁ -C ₆ ) alkanol such as methanol, an aralkyl alcohol such as benzyl alcohol, or an equivalent reactive derivative thereof. ,
The corresponding ester compound (IV) is obtained in each case.

Next, the compound of formula (IV) is dissolved in an organic solvent such as dichloromethane, and trichloromethane is added to the solution in the presence of an organic base such as 1,8-diazabicyclo [5.4.0] -7-undecene. The oxazoline ring is formed by adding acetonitrile and reacting at room temperature. This gives the formula
The compound of (V) is obtained. The compound of formula (V) is dissolved in a mixed solvent of pyridine and water or another suitable solvent, and an acid such as para-toluenesulfonic acid is added to the solution to hydrolyze the compound, whereby the oxazoline compound of formula (V) is hydrolyzed. Open the ring. A compound of formula (VI) is thus obtained.

The compound of the formula (VI) is dissolved in a solvent such as ethanol, a metal hydride such as sodium borohydride is added to the solution, and the mixture is reacted at room temperature. This gives the compound of formula (VII) upon detrichloroacetylation of the compound of formula (VI). The compound of formula (VII) is treated with 1-4 N HCl
-Dissolve in an organic solvent containing hydrogen chloride such as dioxane,
When the elimination reaction of the protecting group is carried out at room temperature, the amino protecting group (R) and the carboxyl protecting group (R ^a ) are removed. This produces the compound No. 1 of the invention of formula (Ia) above as the hydrochloride salt.

The third aspect of the present invention relates to a method for producing the compound of the present invention represented by the above formula (Ib). The third method of the present invention comprises a step of amidinating the 2-position amino group of the compound of formula (VII) starting from the compound of formula (VII) obtained as an intermediate in the method of the second invention. including. That is, specifically, in the third aspect of the present invention, the following formula (VII) (In the formula, R represents an imino protecting group and R ^a represents a carboxyl protecting group which is an ester forming group).
R, 4S, 5S, 6R) -6-acetamido-4-amino-5
-Hydroxypiperidine-3-carboxylic acid protected derivative (Wherein A represents an amino-protecting group which is an alkoxycarbonyl group, an aryloxycarbonyl group or an aralkyloxycarbonyl group), and amidinized by reacting with N, N′-di-protected-thiourea Formula (VII
I) (Wherein R, R ^a and A have the same meanings as described above) and then the respective protecting groups (R, R ^a and A) are eliminated from the compound of formula (VIII). Which is characterized by the following equation (Ib) (3R, 4S, 5S, 6R) -6-acetamide
A method for producing -4-guanidino-5-hydroxypiperidine-3-carboxylic acid, and salts thereof is provided.

A method for carrying out each step of the third method of the present invention will be described below. In this method, in the step of reacting the compound of formula (VII) with N, N′-di-protected-thiourea as described above to amidinate, the amidinization reaction of the compound of formula (VII) is performed by , N-dimethylformamide or the like in an organic solvent in the presence of an organic base such as triethylamine,
It may be carried out by adding N, N'-di-protected-thiourea and mercuric chloride, cooling the mixture, and reacting with stirring. This gives the formula (VII
The compound of I) is obtained. The reaction for obtaining the compound of the formula (VIII) from the compound of the formula (VII) can also be carried out by a general guanidination reaction.

Then, the compound of formula (VIII) is added to 1 to 4 NH
It is dissolved in an organic solvent containing hydrogen chloride such as Cl-dioxane, and this solution is subjected to a reaction for eliminating a protecting group at room temperature. Thereby, the amino protecting group (R), the guanidino group protecting group (A) and the carboxyl protecting group (R ^a ) are eliminated from the compound of formula (VIII) to give the compound of the present invention of formula (Ib) above. No. 2
Is produced as the hydrochloride salt.

The fourth aspect of the present invention relates to a process for producing the compound of the present invention represented by the above formula (Ic). In this method, the compound of the general formula (I
X) (3R, 4S, 5R, 6S) -N-protected-6-amino-3-hydroxymethyl-4,5-dihydroxy-4,5-
O-di protected piperidine (see Japanese Patent Application No. 7-250142)
Of the formula (Ic) included in the compound of the general formula (I) according to the present invention by preparing a compound of formula (IX) and chemically converting the compound in a plurality of steps using the compound of formula (IX) as a starting compound.
To produce (3R, 4S, 5S, 6R) -4-amino-5-hydroxy-6-trifluoroacetamidopiperidine-3-carboxylic acid (Compound No. 3).

That is, specifically, in the fourth aspect of the present invention, the following formula (I
X) (Wherein R represents an imino protecting group, X ¹ and X ² are each a monovalent hydroxyl protecting group, or both represent a divalent hydroxyl protecting group together) (3R , 4S, 5R, 6S) -N-Protected-6-amino-3
-Hydroxymethyl-4,5-dihydroxy-4,5-O-di
-Protection-The 6-amino group of piperidine is protected by an amino protecting group (R ^b ) which can be eliminated by a different elimination method from the imino protecting group (R), and the following formula (X) (Wherein R, R ^b , X ¹ and X ² have the same meanings as described above), and then the 3-hydroxymethyl group of the compound of formula (X) is oxidized to a carboxyl group. Convert to the following equation (XI) (Wherein R and R ^b and X ¹ and X ² have the same meanings as defined above) to produce a carboxylic acid compound, and then reacting the compound of formula (XI) with an esterifying agent (XI
I) (In the formula, R, X ¹ and X ² have the same meanings as described above, R ^a represents a carboxyl-protecting group which is an ester-forming group, and R ^b represents an amino-protecting group which can be eliminated by an elimination method different from R. Represent)
The ester compound of formula (XII) is formed, and then the 4-position hydroxyl group of the compound of formula (XII) is eliminated (deoxygenation) and the bond between the 3- and 4-position carbons is converted to a double bond (enol). Then, the following equation (XIII) A compound of formula (wherein R, ^Ra and ^Rb have the same meanings as described above) is formed, and then the compound of formula (XIII) is reacted with trichloroacetonitrile to form an oxazoline ring. Formula (XIV) (Wherein R, ^Ra and ^Rb have the same meanings as described above), and then the compound of formula (XIV) is hydrolyzed to open the oxazoline ring of the compound of formula (XIV). Ring, which gives the following formula (XV) (Wherein R, R ^a and R ^b have the same meanings as described above), and the trichloroacetyl group is eliminated from the compound of formula (XV) to give the following formula (XVI) (Wherein R, ^Ra and ^Rb have the same meanings as described above), and the 4-position amino group of the compound of formula (XVI) is protected with the same amino group as the imino protecting group (R). Protected by a group of formula (XVII) (Wherein R, R ^a and R ^b have the same meanings as described above), and the carboxyl protecting group (R ^a ) is eliminated from the compound of formula (XVII) to give the following formula (XVII- a) A method of producing a compound represented by the formula (wherein R and R ^b have the same meaning as described above), and removing the imino protecting group (R) from the 3-position carboxyl group of the compound of formula (XVII-a) Protected by a carboxyl protecting group (R ^c ) which can be eliminated in the same reaction as in the following formula (XVIII) (Wherein, R and R ^b have the same meanings as defined above, R ^c is a carboxyl protecting group) to produce a compound represented by the following amino protecting group from the compound of formula (XVIII) and (R ^b) Desorption and the following formula (XVIII-a) (Wherein R and R ^c have the same meanings as described above), and the amino group at the 6-position of the compound of formula (XVIII-a) is trifluoroacetylated to give the following formula (XIX) (Wherein R and R ^c have the same meanings as described above), and then the amino protecting group (R) and the carboxyl protecting group (R ^c ) are removed from the compound of formula (XIX). The following equation (Ic) characterized by (3R, 4S, 5S, 6R) -4-amino-5-
Hydroxy-6-trifluoroacetamide piperidine-
Methods for producing 3-carboxylic acids, and salts thereof, are provided.

Next, a method for carrying out each step of the fourth method of the present invention will be described. General formula (IX) used as a starting compound
Are novel compounds (3R, 4S, 5R, 6
S) -6-Amino-N- (tertiary butoxycarbonyl)-
3-hydroxymethyl-4,5-dihydroxy-4,5-O-
Isopropylidene-piperidine (see Japanese Patent Application No. 7-250142, filed on Sep. 5, 1995) was subjected to acid hydrolysis, whereby the following formula (A) To obtain (3R, 4S, 5R, 6S) -6-amino-3-hydroxymethyl-4,5-dihydroxy-piperidine, and this compound is provided with an imino protecting group (R) and a hydroxyl protecting group (X ¹ , X ² ) can be prepared by introducing a conventional functional group protection method.

In the compound of formula (XI), the imino protecting group which is R is a tertiary butoxycarbonyl group (CH ₃ ) ₃ C
It is convenient that it is O-CO- (abbreviation: Boc). As the imino-protecting group (R), in general, a commonly used imino-protecting group can be introduced by a protecting method known in sugar chemistry instead of the Boc group. For example, an alkoxycarbonyl group, an aryloxycarbonyl group or an aralkyloxycarbonyl group can be used as a suitable imino protecting group (R) which can be eliminated by a hydrolysis method.

Suitable divalent hydroxyl protecting groups (X ¹ , X ² ) which simultaneously protect the hydroxyl groups at the 4 and 5 positions of the compound of formula (IX) are conveniently isopropylidene groups, Generally, various other groups can be used.
That is, [Wherein Y and Z may be the same or different from each other, and each is hydrogen, an alkyl group (preferably a lower alkyl group having 1 to 4 carbon atoms) or an aryl group (preferably a phenyl group or a substituted phenyl group, for example, para- And a divalent hydroxyl protecting group (including an isopropylidene group). Alternatively, it may be convenient to protect the 4- and 5-position hydroxyl groups simultaneously with a benzylidene group or with a cycloalkylidene group, such as a cyclohexylidene group or a tetrahydropyranylidene group.

The introduction of such a divalent hydroxyl protecting group follows the conventional techniques of hydroxy group protection in sugar chemistry to simultaneously protect two of the hydroxyl groups bound to two adjacent carbon atoms. Done. Alternatively, a trialkylsilyl group, especially a tert-butyldimethylsilyl group, can be used as a suitable monovalent hydroxyl protecting group and can be introduced by a conventional technique.

To protect the hydroxyl groups at the 4- and 5-positions of the compound of formula (A), 4,5-O-as a protecting group.
As an example of the reaction for introducing an isopropylidene group, the compound of the formula (A) is dissolved in an organic solvent such as N, N-dimethylformamide, and the solution is mixed with, for example, 2,2-dimethoxypropane (the isopropylidene group-introducing agent). ) Is added, and a chloride alkylsilane such as chlorotrimethylsilane, or an acid catalyst such as para-toluenesulfonic acid is added, and the mixture is reacted at room temperature for several hours to give a compound of formula (IX)
As an example of the compound represented by, a 4,5-O-diisopropylidene derivative is obtained.

When 2,2-dimethoxypropane used in the above-mentioned hydroxyl group protection reaction is replaced by benzaldehyde dimethyl acetal or 1,1-dimethoxycyclohexane or other various acetal and ketal reagents, 4 A compound of the formula (IX) is obtained which is loaded with a 5-O-benzylidene group or a cyclohexylidene group or other acetal and ketal groups.

In the first step of the method of the fourth aspect of the present invention, an amino-protecting group which can be eliminated by an elimination method in which the 6-position amino group of the compound of formula (IX) used as a starting compound is different from the imino-protecting group (R) Protected by (R ^b ). The amino protecting group (R ^b ) is conveniently an aralkyloxycarbonyl group, preferably a benzyloxycarbonyl group, which can be eliminated by hydrolysis. The amino protecting group (R ^b ) can be introduced by a conventional method. For example, a compound of formula (IX) is dissolved in an organic solvent such as methanol, and aralkyloxycarbonyl chloride, preferably benzyloxycarbonyl chloride, is added to the solution in the presence of a base such as N, N-diisopropylamine to obtain a solution. By cooling and reacting the resulting mixture,
A compound of formula (X) is obtained in which the 6-position amino group is protected.

Next, the 3-hydroxymethyl group of the compound of formula (X) is oxidized to a carboxyl group. For this the formula
The compound (X) is dissolved in a mixed solvent of carbon tetrachloride, acetonitrile and water, an oxidizing agent, preferably ruthenium dioxide, is added to the solution in the presence of sodium metaperiodate, and the resulting mixture is subjected to an oxidation reaction at room temperature. I do. This gives the free carboxylic acid compound of formula (XI).

The compound of formula (XI) is dissolved in a mixed solvent of methylene chloride and methanol, an esterifying agent is added to the solution, and the carboxyl group of the compound of formula (XI) is protected by esterification at room temperature. This gives the compound of formula (XII). The esterifying agent can be the same as that used in the second method of the present invention.

Next, the 4-hydroxyl group of the compound of formula (XII) is eliminated and enolized. For this purpose, the compound is dissolved in an organic solvent such as tetrahydrofuran, and an alkoxyalkali metal, preferably potassium tertiary butoxide is added to the solution, and the reaction is carried out under cooling. This gives the compound of formula (XIII).

Further, a reaction for forming an oxazoline ring between the 5-position hydroxyl group and the 4-position of the compound of formula (XIII) is carried out. For this purpose, the compound is dissolved in an organic solvent such as benzene, and 1,8-diazabicyclo [5.4.
Trichloroacetonitrile is added in the presence of an organic base such as 0] -7-undecene and reacted at room temperature. Thereby, an oxazoline ring is formed and a compound of formula (XIV) is obtained.

Further, the oxazoline ring of the compound of formula (XIV) is opened. For this purpose, the compound is dissolved in a solvent such as a mixed solvent of pyridine and water, and an acid such as para-toluenesulfonic acid is added to the solution for hydrolysis. This opens the oxazoline ring to give the compound of formula (XV).

Next, the trichloroacetyl group is eliminated from the compound of formula (XV). For this purpose, the compound is dissolved in a solvent such as ethanol, a metal hydride such as sodium borohydride is added to the solution, and the mixture is reacted at room temperature. Thereby, the compound of formula (XV) undergoes detrichloroacetylation to obtain the compound of formula (XVI).

Further, the 4-position amino group of the compound of formula (XVI) is protected with the same protecting group as the imino protecting group (R). For this purpose, the compound is dissolved in a solvent such as methanol, and an amino-protecting group-introducing reagent, preferably di-tertiary carbonate, is added to the solution in the presence of a base such as N, N-diisopropylamine. React. By carrying out protection of the 4-position amino group, a compound of formula (XVII) is obtained.

Further, in order to eliminate the carboxyl protecting group (R ^a ) of the compound of the formula (XVII), the compound is dissolved in a solvent such as methanol, and 1M aqueous sodium hydroxide solution is added to the solution to give an alkaline hydrolyzate. When decomposed, the formula (XV
A deesterified product of II-a) is obtained. An ester for dissolving a compound of the formula (XVII-a) in a solvent such as methylene chloride and introducing into the solution a carboxyl protecting group (R ^c ) of a kind that can be eliminated by the same elimination method as the imino protecting group (R). Agent, N,
The re-esterification reaction is carried out in the presence of a base such as N-diisopropylamine. This gives the compound of formula (XVIII).

Next, the protecting group (R ^b ) is removed from the 6-position amino group of the compound of formula (XVIII). For this purpose, the compound is dissolved in a solvent such as acetonitrile and 10% is added to the solution.
Palladium on carbon is added, and hydrogenolysis is performed at room temperature in a hydrogen atmosphere. This gives the compound of formula (XVIII-a). Trifluoroacetylation of the 6-amino group of the resulting compound of formula (XVIII-a) is then carried out. Therefore, the compound is dissolved in a solvent such as methylene chloride, and a trifluoroacetylating agent, preferably trifluoroacetic anhydride, is added to the solution in the presence of a base such as pyridine, and the reaction is conducted. Trifluoroacetylation is performed. This gives the compound of formula (XIX).

Finally, a reaction for eliminating the amino protecting group (R) and the carboxyl protecting group (R ^c ) from the compound of the formula (XIX) is carried out. To this end, the compound is
N hydrochloric acid gas-dissolving in an organic solvent containing hydrogen chloride such as dioxane and removing the amino protecting group (R) and the carboxyl protecting group (R ^c ) by acid hydrolysis, the compound of the present invention of formula (Ic) No. 3 is produced as the hydrochloride salt.

Furthermore, the fifth aspect of the present invention is based on the above formula (Id)
Of the present invention. A compound of formula (Id), ie (3R, 4S, 5S, 6R) -4-guanidino-5-hydroxy-6-trifluoroacetamidopiperidine-3-
The carboxylic acid (Compound No. 4) can be prepared by subjecting it to an amidinolysis reaction starting from the compound of formula (XVI) obtained as an intermediate in the method of the fourth aspect of the present invention.

That is, in the fifth aspect of the present invention, the following formula (XVI) [In the formula, R represents an imino protecting group, R ^a represents a carboxyl protecting group which is an ester forming group, and R ^b represents an amino protecting group which can be eliminated by a different elimination method from the imino protecting group (R)]. Indicated by (3R, 4S, 5S, 6R or 6
The protected derivative of (S) -6-N-protected-4,6-diamino-5-piperidine-3-carboxylic acid is represented by the following formula (Wherein A represents an amino-protecting group which is an alkoxycarbonyl group, an aryloxycarbonyl group or an aralkyloxycarbonyl group), and is N, N'-di-protected.
-The compound of the following formula (XX) (In the formula, R, ^Ra , ^Rb and A have the same meanings as described above.)
The compound represented by the formula (XX) is then removed by selectively removing the amino-protecting group (R ^b ) at the 6-position of the compound of the formula (XX).
-a) (Wherein R, R ^a, and A have the same meanings as described above), and then the amino group at the 6-position of the compound of formula (XX-a) is trifluoroacetylated to Formula (XXI) (Wherein R, R ^a and A have the same meanings as described above), and the respective protecting groups (R, R ^a and A) are removed from the compound of formula (XXI). The following formula (Id) characterized by (3R, 4S, 5S, 6R) -4-guanidino-
Methods for making 5-hydroxy-6-trifluoroacetamidopiperidine-3-carboxylic acid, and salts thereof are provided.

A method for carrying out each step of the fifth method of the present invention will be described below. In this method, the compound of formula (XVI) is first prepared by reacting the compound of formula (XVI) in an organic solvent such as N, N-dimethylformamide in the presence of an organic base such as triethylamine.
N'-di-protected-thiourea and mercuric chloride are added and reacted under cooling. This gives the compound of formula (XX). The amino protecting group of the above thiourea derivative is generally an alkyloxycarbonyl group, an aryloxycarbonyl group, an aralkyloxycarbonyl group or the like, which is a commonly used protecting group for urea, as in the method of the third aspect of the present invention. Available. The reaction for obtaining the compound of the formula (XX) from the compound of the formula (XVI) can also be carried out by a general guanidination reaction.

Next, the protecting group (R ^b ) for the amino group at the 6-position of the compound of formula (XX) is selectively eliminated. For this purpose, the compound is dissolved in a solvent such as acetonitrile and the solution is subjected to the elimination reaction of the amino protecting group (R ^b ), whereby the compound of formula (XX-
The compound of a) is obtained. The 6-position amino group of the resulting compound of formula (XX-a) is then trifluoroacetylated. For this purpose, the compound is dissolved in a solvent such as methylene chloride, and a trifluoroacetylating agent, preferably trifluoroacetic anhydride, is added to the solution and reacted in the presence of a base such as pyridine. The amino group at the 6-position is trifluoroacetylated to give a compound of formula (XXI).

Finally, a reaction for eliminating the protecting groups (R, R ^a and A) from the compound of formula (XXI) above is carried out. For this purpose, it is convenient to dissolve the compound of formula (XXI) in an organic solvent containing hydrogen chloride such as 1-4N HCl-dioxane and subject it to acid hydrolysis. Thereby an amino protecting group (R),
And the carboxyl protecting group (R ^a ) and the guanidino protecting group (A) are eliminated to give a compound N of the formula (Id) of the present invention.
o.4 is produced as the hydrochloride salt.

In the second to fifth methods of the present invention, the formulas (Ia) and (I
When the compound (b), (Ic) or (Id) is obtained in the form of an acid addition salt such as a hydrochloride as described above, an aqueous solution of the acid addition salt is prepared by a conventional method in a cation exchange resin such as Dow. Compound No. 1, No. 1 of the invention in the form of the free base, either by treatment with X50W (Dow Chemical Company, USA) (H ⁺ form) or by purification by means of a solvent system containing ammonia. .2, No. 3 or No. 4 is obtained.

According to the second method of the present invention, 4-deoxy-
Starting from 3-en-1-N- (tert-butoxycarbonyl) siastatin B, (3R, 4S, 5S, 6R) -6-acetamido-4-amino-5 of the formula (Ia) according to the invention. The method for producing -hydroxypiperidine-3-carboxylic acid (Compound No. 1) can be carried out, for example, by a reaction route consisting of a plurality of steps shown in the following Reaction Chart 1. However, in Reaction Chart 1, Boc represents a tertiary butoxycarbonyl group (CH ₃ ) ₃ CO-CO- as an imino protecting group,
Ph represents a phenyl group.

Reaction Chart 1 That is, to explain the Reaction Chart 1, 4-deoxy-3-ene-1-N- (tert-butoxycarbonyl) siastatin B represented by the formula (III-1) was added to a mixed solvent of dichloromethane and methanol. After dissolution, diphenyldiazomethane is added to the solution and reacted at room temperature with stirring to esterify the carboxyl group of the compound of formula (III-1) with diphenylmethyl group -CHPh ₂ . This gives the compound of formula (IV-1).

The compound of formula (IV-1) was dissolved in dichloromethane, and 1,8-diazabicyclo [5.4.0] -7 was added to the solution.
-Oxazoline ring formation by addition of undecene and trichloroacetonitrile and reaction at room temperature gives compounds of formula (V-1). Furthermore, the formula (V-
When the compound of 1) is dissolved in a mixed solvent of pyridine and water and para-toluenesulfonic acid is added to the solution for hydrolysis to open the oxazoline ring, the compound of formula (VI-
The compound of 1) is obtained.

When the compound of the formula (VI-1) is dissolved in ethanol, sodium borohydride is added to the solution, and the reaction is carried out at room temperature to detrichloroacetylate the 4-position amino group. The compound of 1) is obtained.

The compound of the above formula (VII-1) is added to 1 to 4N
When the compound is dissolved in HCl-dioxane and the protecting group is eliminated at room temperature, the amino protecting group Boc and the carboxyl protecting group diphenylmethyl group are removed, and the compound No. 1 of the present invention of the formula (Ia) is converted into a hydrochloride salt. To generate.

According to the third method of the present invention, starting from the compound of the formula (VII-1) shown in the above reaction chart 1,
The compound represented by the formula (Ib) according to the present invention (3R, 4S, 5S, 6
The method for producing R) -6-acetamido-4-guanidino-5-hydroxypiperidine-3-carboxylic acid (Compound No. 2) is carried out, for example, by a reaction route consisting of a plurality of steps shown in the following Reaction Chart 2. it can. However, in Reaction Chart 2, Boc
Represents a tertiary butoxycarbonyl group as an imino protecting group, and Ph represents a phenyl group.

Reaction Chart 2 That is, to explain the above Reaction Chart 2,
The compound (VII-1) is dissolved in N, N-dimethylformamide, and the solution is mixed with an organic base such as triethylamine and N,
N'-di- (tert-butyloxycarbonyl) thiourea and mercuric chloride are added and the reaction is carried out under cooling, for example, at 0 ° C. This gives the compound of formula (VIII-1). Then, the compound of formula (VIII-1) is dissolved in 1 to 4N HCl-dioxane, and the compound of formula (VIII-1) is subjected to acid hydrolysis to give an amino protecting group (Boc) and a carboxyl protecting group.
The elimination reaction with (-CHPh ₂ ) is performed. This gives the formula (I
Inventive compound No. 2 of b) is produced as the hydrochloride salt.

According to the fourth method of the present invention, the following formula (IX-
1) starting from (3R, 4S, 5R, 6S) -6- (N-tert-butoxycarbonylamino) -3-hydroxymethyl-4,5-dihydroxy-4,5-O-isopropylidene-piperidine To produce (3R, 4S, 5S, 6R) -4-amino-5-hydroxy-6-trifluoroacetamidopiperidine-3-carboxylic acid of formula (Ic) according to the present invention (Compound No. 3). The method can be carried out, for example, in a reaction process consisting of a plurality of steps shown in the following Reaction Chart 3.

However, in Reaction Chart 3, Boc and Ph have the same meanings as described above, Me represents a methyl group, and Z represents a benzyloxycarbonyl group C ₆ H ₅ CH as an amino protecting group.
₂ -O-CO-, MEM is a 2-methoxyethoxymethyl group CH ₃ OCH ₂ CH ₂ OCH which is a carboxyl protecting group
Indicates _2- .

Reaction Chart 3 That is, in order to explain the above Reaction Chart 3, the formula
(3R, 4S, 5R, 6S) -6-amino represented by (XI-1)
-1-N- (tert-butoxycarbonyl) -4,5-dihydroxy-3-hydroxymethyl-4,5-O-isopropylidenepiperidine was dissolved in methanol and N, N-diisopropylamine and benzyl chloride were added to the solution. Oxycarbonyl is added and the reaction is carried out under cooling, for example at 0 ° C. This gives the compound of formula (X-1) in which the amino group at the 6-position is protected.

The compound of the formula (X-1) is dissolved in a mixed solvent of carbon tetrachloride, acetonitrile and water, sodium metaperiodate and ruthenium dioxide are added to the solution, and the mixture is reacted at room temperature to obtain the compound of the formula (X-1) The compound of formula (XI-1) is obtained by oxidizing the 3-hydroxymethyl group of the compound of The compound of formula (XI-1) is dissolved in a mixed solvent of methylene chloride and methanol, diphenyldiazomethane is added to the solution, and the mixture is stirred at room temperature to carry out an esterification reaction in which the 6-position carboxyl group is protected with a diphenylmethyl group. . This gives the compound of formula (XII-1).

The compound of the formula (XIII-1) is dissolved in tetrahydrofuran, potassium tert-butoxide is added to the solution, and the elimination reaction and the enolization reaction are carried out under cooling at, for example, 0 ° C. The compound of -1) is obtained. The compound of formula (XIII-1) is dissolved in benzene, 1,8-diazabicyclo [5.4.0] -7-undecene and trichloroacetonitrile are added to the solution, and the mixture is reacted at room temperature. This provides a compound of formula (XIV-1) upon forming an oxazoline ring.

The compound of formula (XIV-1) is dissolved in a mixed solvent of pyridine and water, and para-toluenesulfonic acid is added to the solution for hydrolysis. This opens the oxazoline ring to give the compound of formula (XV-1). Expression (XV-
The compound of 1) is dissolved in ethanol, sodium borohydride is added to the solution, and the mixture is reacted at room temperature. Thereby, when the trichloroacetyl group is eliminated from the compound of formula (XV-1), the compound of formula (XVI-1) is obtained.

A reaction of dissolving the compound of the formula (XVI-1) in methanol, adding N, N-diisopropylamine and ditertiary carbonate to the solution, and protecting the 3-position amino group with the Boc group at room temperature. I do. This allows
A compound of formula (XVII-1) is obtained. The compound of formula (XVII-1) is dissolved in methanol, 1 M aqueous sodium hydroxide solution is added to the solution, and alkali hydrolysis is performed to obtain a free carboxylic acid compound of formula (XVII-a-1). The obtained compound (XVII-a-1) is dissolved in methylene chloride, and N, N-diisopropylamine and chloromethyl (2-methoxy) ethyl ether are added to the solution to react. Thus, when the carboxyl group is re-esterified with the MEM group, the compound of formula (XVIII-a-1) is obtained.

The compound of formula (XVIII-a-1) was dissolved in acetonitrile, 10% palladium-carbon was added to the solution, and hydrogenolysis was carried out under stirring in a hydrogen atmosphere at room temperature to debenzyloxycarbonylate. To give a compound of formula (XVIII-a-1). The compound of formula (XVIII-a-1) is dissolved in a methylene chloride solvent, pyridine and trifluoroacetic anhydride are added to the solution, and the mixture is reacted with stirring. By performing trifluoroacetylation of the 6-position amino group, the formula (XI
The compound of X-1) is obtained.

The compound of the above formula (XIX-1) is added to 1 to 4N
It is dissolved in HCl-dioxane and subjected to elimination reaction of the protecting group at room temperature. As a result, the amino protecting group Boc and the 2-methoxyethoxymethyl group (-ME
Removal of M) produces an inventive compound No. 4 of formula (Id) as the hydrochloride salt.

According to the fifth method of the present invention, starting from the compound of formula (XVI-1) shown in Reaction Chart 3, a compound of formula (Id) according to the present invention (compound No. 4) is obtained. The production method can be carried out, for example, in a reaction process including a plurality of steps shown in the following reaction chart 4. However, in Reaction Chart 4, Boc, Ph and Z have the same meanings as described above.

Reaction Chart 4 To explain Reaction Chart 4, the compound of formula (XVI-1) was dissolved in N, N-dimethylformamide, and triethylamine and N, N'-di- (tert-butoxycarbonyl) thiourea and mercuric chloride were added. In addition, the reaction is carried out at 0 ° C. with stirring. This gives the compound of formula (XX-1). The compound of formula (XX-1) is dissolved in acetonitrile, 10% palladium carbon is added to the solution, and the debenzyloxycarbonylation reaction is carried out under stirring in a hydrogen atmosphere at room temperature. This gives the compound of formula (XX-a-1). The obtained compound of formula (XX-a-1) is dissolved in methylene chloride, pyridine and trifluoroacetic anhydride are added to the solution, and the mixture is reacted under stirring. As a result, trifluoroacetylation of the 6-position amino group is performed to obtain the compound of formula (XXI-1).

The compound of the above formula (XXI-1) was added to 1 to 4N
It is dissolved in HCl-dioxane and the elimination reaction of the protecting group is carried out at room temperature with stirring. As a result, when the amino protecting group Boc and the carboxyl protecting group diphenylmethyl group are removed, the compound No. 4 of the formula (Id) of the present invention is produced as a hydrochloride.

The compound of the general formula (IX) used as the starting compound in the fourth method of the present invention is a novel compound,
The manufacturing method of this is demonstrated below.

The compound of general formula (IX) is summarized in the following steps: (In the formula, R is an imino protecting group having the same meaning as described above, preferably an alkoxycarbonyl group).
The hydroxyl groups at positions 4 and 5 of cystatin B are
Protecting with a monovalent or divalent hydroxyl protecting group (X ¹ , X ² ), the following general formula (C) (In the formula, R is an imino protecting group, and X ¹ and X ² are 1
A divalent hydroxyl protecting group, or both X ¹ and X ² together represent one divalent hydroxyl protecting group, such as isopropylidene, benzylidene, cycloalkylidene or tetrahydropyranylidene.] A step of producing the represented N-protected-4,5-di-O-protected cystatin B, and reacting the 3-position carboxyl group of the compound of formula (C) with an esterifying agent ) (Wherein R, X ¹ and X ² have the same meanings as described above, and T is an ester-forming group), and a step of producing a protected cystatin B derivative, and a 3-position carboxy of the compound of formula (D) The rate group —CO ₂ T is converted into a hydroxymethyl group by reduction to give the following general formula (E) (Wherein R, X ¹ and X ² have the same meaning as described above) (3R, 4S, 5R, 6R or 6S) -6-acetamido-N-protected-3-hydroxymethyl-4,5 -Dihydroxy
-4,5-O-diprotected-piperidine is formed, and further, a free amino group is formed from the acetamide group at the 6-position of the compound of the general formula (E) by deacetylation to give the following general formula (F ) (Wherein R, X ¹ and X ² have the same meanings as described above) (3R, 4S, 5R, 6S) -1-N-protected-6-amino-
3-hydroxymethyl-4,5-dihydroxy-4,5-O-
It can be produced by a method comprising the step of producing diprotected-piperidine (see Reference Example 1 below).

From the compound of the above formula (F), an imino protecting group (R) and a hydroxyl protecting group (X ¹ , X ² ) can be prepared by a conventional method.
Is eliminated, the compound of the above formula (A), namely (3
R, 4S, 5R, 6S) -6-Amino-3-hydroxymethyl-4,5-dihydroxypiperidine can be obtained. Further, if desired, by introducing an arbitrary imino protecting group and hydroxyl protecting group into the compound of the formula (A) by a conventional method,
Compounds of general formula (IX) can be prepared.

N-protected-siastatin B of formula (B) above
In, it is convenient that the imino protecting group which is R is a tertiary butoxycarbonyl group (abbreviation: Boc).

When a 4,5-O-isopropylidene group is introduced as a protecting group in order to protect the hydroxyl groups at the 4- and 5-positions of N-protected-cystatin B of the formula (B), the formula ( B) The compound is dissolved in an organic solvent such as N, N-dimethylformamide, and the solution is dissolved in, for example, 2,2-
By adding dimethoxypropane (an isopropylidene group-introducing agent) and further adding an acid catalyst such as chloroalkylsilane such as chlorotrimethylsilane or paratoluenesulfonic acid, and reacting at room temperature for several hours, 5-O-di-protected-siastatin B is obtained, provided that X ¹ and X ² together represent an isopropylidene group.

If benzaldehyde dimethyl acetal or 1,1-dimethoxycyclohexane or other various acetal and ketal reagents are used in place of the above 2,2-dimethoxypropane, 4,5-O-
A compound of the general formula (C) having a benzylidene group or a cyclohexylidene group or other acetal and ketal groups introduced therein is obtained.

Next, in order to introduce an ester-forming group (T) into the 3-position carboxyl group of the compound of the general formula (C),
The compound is dissolved in an organic solvent such as N, N-dimethylformamide, a base such as diisopropylamine or triethylamine is added to the solution, and, for example, methoxyethoxymethyl chloride is added as an esterifying agent in the presence of the base. By reacting the obtained reaction mixture at room temperature for several hours, an esterified compound of the general formula (D) (where T is a methoxyethoxymethyl group) is obtained.

As the above-mentioned esterifying agent, alkyl chloride instead of methoxyethoxymethyl chloride,
Aryl chloride, aralkyl chloride, alkoxide-substituted lower alkoxymethyl chloride such as lower alkoxymethyl chloride, aralkyloxymethyl chloride, methoxymethyl chloride, etc. are ester-forming groups.
When used as an introducing agent of (T), the ester-forming group (T) in the compound of formula (D) is an alkyl group, an aryl group, an aralkyl group, a lower alkoxymethyl group, an aralkyloxymethyl group or an alkoxy-substituted-lower group, respectively. A compound of formula (D) is produced when it is an alkoxymethyl group.

Then, in order to reduce the 3-position carboxylate group (--CO ₂ T) of the compound of the general formula (D), the compound is added to a single organic solvent such as tetrahydrofuran or a mixed organic solvent thereof. After dissolution, a reducing agent such as sodium borohydride, lithium aluminum hydride or diisobutylaluminum hydride is added to this solution and reacted at room temperature for several hours, the 3-position carboxylate group is converted to a hydroxymethyl group by reduction. . Thereby, the alcohol compound of the general formula (E) is obtained.

Then, in order to convert the 6-position acetamide group of the compound of the general formula (E) into a free amino group, the compound is dissolved in hydrazine hydrate, for example, 50 to 100.
When the reaction is carried out at ℃ for 7 to 14 days, a compound of the general formula (F) having an amino group released by elimination of the acetyl group from the acetamide group is obtained.

[0092]

Embodiments of the present invention will be described below. Example 1 is a compound of formula (Ia) according to the invention, Example 2 is a compound of formula
Example 3 shows an example of the preparation of the compound of formula (Ib), Example 3 is the compound of formula (Ic) and Example 4 is the compound of formula (Id). However, these examples are merely illustrative and do not limit the invention. Various variations and modifications are possible within the scope of the invention.

Example 1 (1) (5R, 6S) -acetamido-1-N- (tert-butoxycarbonyl) -3,4-didehydropiperidine-
3-carboxylic acid diphenylmethyl ester [formula (IV-1)
The compound of (5S, 6S) -6-acetamido-1-N- (tert-butoxycarbonyl) -3,4 represented by the above formula (III-1).
-Didehydropiperidine-3-carboxylic acid (1.87 g) was dissolved in 60 ml of a mixed solvent of dichloromethane-methanol (1: 1) and diphenyldiazomethane (3.64 g) was added to the solution.
Was added and stirred overnight at room temperature to carry out an esterification reaction.
After acetic acid (0.9 ml) was added to stop the reaction, the reaction solution was stirred at room temperature for 1 hour, and the solvent was evaporated under reduced pressure. The residue was dissolved in chloroform and washed 3 times with saturated aqueous sodium hydrogen carbonate solution. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was evaporated under reduced pressure.
The residue was subjected to silica gel column chromatography (elution solvent: dichloromethane-methanol, 99: 1 to 98:
2, gradient method), and the title compound (2.66 in the form of a colorless foam) was obtained.
g, 92%).

[Α] ²⁴ _D + 90.7 ° (c 0.86, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, δppm): 1.49 (9 H, s, CO
OC (CH ₃ ) ₃ ), 1.89 (3H, s, NHCOC H ₃ ), 3.86 (1H, broad d,
J = 19.1 Hz, 2-Ha), 4.16 (1H, broad, 5-H), 4.52 (1H, d
d, J = 19.1 and 2.0 Hz, 2-Hb), 6.10 (1H, broad s, 6-
H), 6.97 (1H, s, Ph ₂ C H O-), 7.12 (1H, m, 4-H), 7.27-7.
42 (10H, m, Ph).

(2) (3aS, 4R, 7S, 7aS)-
7-acetamido-6-N- (tert-butoxycarbonyl) -2- (trichloromethyl) -4- (diphenylmethyloxycarbonyl) -3a, 7a-dihydrooxazolo [5,4-c] piperidine [formula (V -1) compound] The compound of the formula (IV-1) (233 mg) obtained in the previous section (1) was dissolved in dichloromethane (5 ml), and trichloroacetonitrile (65.2 μl) and DBU (15.0 μl) were added to the solution. )
Was added and the reaction was carried out by stirring at room temperature for 5 minutes. The reaction was stopped by adding saturated ammonium chloride aqueous solution to the reaction solution. The reaction solution was diluted with chloroform and washed with a saturated aqueous solution of ammonium chloride three times. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent for filtration was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: toluene-acetone, 6: 1) to give the title compound (234 mg, 76%) as a colorless foam.

[Α] ²⁴ _D + 20.5 ° (c 0.86, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.47 (9
H, s, COOC (CH ₃ ) ₃ ), 1.75 (3H, s, NHCOC H ₃ ), 2.77 (1H,
dt, J = 8.3, 5.6 Hz, 3-H), 3.46 (1H, dd, J = 13.7,8.3H
z, 2-Hax), 3.89 (1H, dd, J = 13.7, 4.9Hz, 2-Heq), 4.9
6-5.03 (2H, m, 4-H, 5-H), 6.18 (1H, s, 6-H), 7.10 (1H,
_{s, Ph 2 C H O-)} , 7.26-7.42 (10H, m, Ph).

(3) (3R, 4S, 5S, 6S) -6-
Acetamide-1-N- (tert-butoxycarbonyl) -5-hydroxy-4-trichloroacetamide piperidine-3-carboxylic acid diphenylmethyl ester [formula (VI
-1) Compound] The compound of the formula (V-1) (1.53 g) obtained in the above (2) was dissolved in a mixed solvent of pyridine (24 ml) and water (6 ml), and para-toluene was added to the solution. Sulfonic acid (monohydrate, 0.71
3 g, 3.75 mmol) was added and the reaction was carried out by stirring at 80 ° C. for 2 hours. The reaction solution was cooled to room temperature, diluted with chloroform, and washed 3 times with saturated aqueous sodium hydrogen carbonate solution. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. After azeotropically distilling the residue 3 times with toluene, silica gel column chromatography (eluting solvent: chloroform-methanol,
30: 1) to give the title compound as a colorless foam (1.20).
g, 77%).

[Α] ²⁴ _D + 3.8 ° (c 0.95, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.47 (9
H, s, COOC (CH ₃ ) ₃ ), 1.99 (3H, s, NHCOC H ₃ ), 3.17 (1H,
broad t, J = 12.7 Hz, 2-Hax), 3.27 (1H, dt, J = 11.5, 4.
2 Hz, 3-H), 3.82 (1H, broad m, 5-H), 4.30 (1H, broad
d, J = 12.7Hz, 2-Heq), 4.47 (1H, dd, J = 11.5, 2.7 Hz, 4
-H), 6.05 (1H, d, J = 1.5 Hz, 6-H), 6.84 (1H, s, Ph ₂ C H O
-), 7.25-7.38 (10H, m, Ph).

(4) (3R, 4S, 5S, 6S) -6-
Preparation of acetamido-4-amino-1-N- (tertiary butoxycarbonyl) -5-hydroxypiperidine-3-carboxylic acid diphenylmethyl ester [compound of formula (VII-1)] Formula (3) The compound of VI-1) (629 mg) was dissolved in ethanol (13 ml), sodium borohydride (14 mg) was added to the solution, and the mixture was stirred at room temperature for 50 minutes for reaction. The reaction solution was diluted with chloroform and washed twice with saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was subjected to silica gel column chromatography (elution solvent: chloroform-methanol, 9:
The title compound (298 mg, 62)
%).

[Α] ²⁴ _D + 19.7 ° (c 0.78, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.47 (9
H, s, COOC (CH ₃ ) ₃ ), 1.95 (3H, s, NHCOC H ₃ ), 2.88 (1H,
dt, J = 11.4, 4.4 Hz, 3-H), 3.01 (1H, t, J = 13.2Hz, 2H
ax), 3.17 (1H, dd, J = 10.7, 2.4 Hz, 4-H), 3.73 (1H, b
road s, 5-H), 4.27 (1H, dd, J = 13.7, 3.9 Hz, 2-Heq),
6.03 (1H, d, J = 2.0 Hz, 6-H), 6.92 (1H, s, Ph ₂ C H O-),
7.26-7.41 (10H, m, Ph).

(5) (3R, 4S, 5S, 6R) -6-
Acetamide-4-amino-5-hydroxypiperidine-3-
Production of Carboxylic Acid [Compound of Formula (Ia)] The compound of the formula (VII-1) (46 mg) obtained in the preceding section (4)
It was dissolved in M hydrochloric acid-dioxane solution (1.5 ml), and the solution was stirred at room temperature overnight. The elimination reaction of the protecting group was carried out. Diethyl ether was added to the reaction solution in which a colorless solid had precipitated and was suspended, and the mixture was well stirred, and then the precipitated colorless solid was precipitated by centrifugation, and the supernatant was removed. After this, the procedure of suspending the precipitate in diethyl ether and centrifuging and then discarding the supernatant was repeated twice. The obtained precipitate was air-dried and then dried under reduced pressure with a pump to obtain a compound of the formula (Ia) No.
Crude 1 (31 mg) was obtained as a colorless solid.

This crude product was adsorbed on Diaion HP20 resin and eluted with water for further purification. The obtained purified product was azeotropically distilled three times with methanol to give a colorless amorphous solid, and then dissolved in a very small amount of methanol. 1M hydrochloric acid-
A dioxane solution was added to precipitate a colorless solid, diethyl ether was further added, and the mixture was stirred well. The deposited colorless solid was precipitated by centrifugation and the supernatant was removed. After this, the procedure of suspending the precipitate in diethyl ether and centrifuging and then discarding the supernatant was repeated twice. The obtained precipitate was air-dried and then dried under reduced pressure with a pump to obtain the dihydrochloride salt of the title compound, ie, the present compound No. 1 (27 mg, 100%) as a colorless solid.

Example 2 (1) (3R, 4S, 5S, 6S) -6-acetamido-
5- [N, N'-di- (tertiary butoxycarbonyl)
Guanidino] -1-N- (tertiary butoxycarbonyl)
Production of -4-hydroxypiperidine-3-carboxylic acid diphenylmethyl ester [compound of formula (VIII-1)] The compound of formula (VII-1) (242m) obtained in Example 1 (4).
g) in N, N-dimethylformamide (5 ml),
Cooled to 0 ° C. To this solution was added triethylamine (0.28
ml), N, N'-di- (tertiary butoxycarbonyl)
Thiourea (276 mg), mercuric chloride (II) (272 mg)
Was added, and the resulting mixture was stirred at 0 ° C. for 1 hour to carry out an amidinization reaction.

Ethyl acetate was added to the reaction solution in which an ocher-colored solid had precipitated, and the mixture was centrifuged to precipitate the solid, and the supernatant was transferred separately. After this, the precipitate is suspended in ethyl acetate,
After centrifugation, the operation of transferring the supernatant separately was repeated twice. The supernatants were combined and washed twice with water and once with a small amount of saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was azeotroped with toluene three times, and then purified by silica gel column chromatography (eluting solvent: toluene-acetone, 3: 1) to obtain the title compound (320 mg, 88%) as a colorless amorphous solid.

[Α] ²³ _D -2.3 ° (c 0.72, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.47,
1.48, 1.49 (9H, s, COOC (CH ₃ ) ₃ ), 2.01 (3H, s, NHCOC
H ₃ ), 2.99 (1H, dt, J = 11.7, 3.9 Hz, 3-H), 3.16 (1H, m,
2-Hax), 3.73 (1H, m, 5-H), 4.17 (1H, broad d, J = 13.
7 Hz, 2-Heq), 4.68 (1H, dd, J = 2.5, 11.7 Hz, 4H), 6.0
2 (1H, d, J = 2.0 Hz, 6-H), 6.81 (1H, s, Ph ₂ C H O-), 7.24
-7.34 (10H, m, Ph).

(2) (3R, 4S, 5S, 6R) -6-
Acetamide-4-guanidino-5-hydroxypiperidine
-Preparation of 3-carboxylic acid [compound of formula (Ib)] The compound (73 mg) of formula (VIII-1) obtained in the previous section (1) was added to 4
It was dissolved in M hydrochloric acid-dioxane solution (2.5 ml), and the solution was stirred overnight at room temperature to carry out a deprotection reaction. Diethyl ether was added to the reaction solution in which a colorless solid had precipitated and was suspended, and the mixture was well stirred, and then the precipitated colorless solid was precipitated by centrifugation, and the supernatant was removed. After this, the procedure of suspending the precipitate in diethyl ether and centrifuging and then discarding the supernatant was repeated twice. The obtained precipitate was air-dried and then dried under reduced pressure with a pump to obtain a crude product (37 mg) of the compound of formula (Ib) as a colorless solid.

The crude product was adsorbed on Diaion HP20 resin and eluted with water for further purification. The obtained purified product was azeotropically distilled three times with methanol to give a colorless amorphous solid, and then dissolved in a very small amount of methanol. 1M hydrochloric acid-
A dioxane solution was added to precipitate a colorless solid, diethyl ether was further added, and the mixture was stirred well. The deposited colorless solid was precipitated by centrifugation and the supernatant was removed. After this, the procedure of suspending the precipitate in diethyl ether and centrifuging and then discarding the supernatant was repeated twice. The obtained precipitate was air-dried and then dried under reduced pressure with a pump to obtain the dihydrochloride salt (31 mg, 94%) of the title compound, ie, the present compound No. 2 as a colorless solid.

Example 3 (1) (3R, 4S, 5R, 6S) -6-Benzyloxycarbonylamino-1-N- (tert-butoxycarbonyl) -3-hydroxymethyl-4,5-dihydroxy-
Production of 4,5-O-isopropylidenepiperidine [compound of formula (X-1)] The compound represented by the above formula (IX-1) (3R, 4S, 5R, 6)
S) -1-N- (tert-butoxycarbonyl) -6-amino-3-hydroxymethyl-4,5-dihydroxypiperidine (3.24 g) was dissolved in methanol (65 ml) and the solution was cooled to 0 ° C. . To this, N, N-diisopropylethylamine (7.46 ml) and benzyloxycarbonyl chloride (4.58 ml) were added, and the mixture was stirred at 0 ° C for 1 hr. Then, benzyloxycarbonyl chloride (0.76 ml) was added to the reaction solution, and the mixture was stirred at 0 ° C. for 30 minutes to carry out N-benzyloxycarbonylation reaction. Water (1.5m
l) was added to stop the reaction, and the obtained reaction solution was stirred at room temperature for 30 minutes. Thereafter, the solvent was distilled off under reduced pressure.

The residue was dissolved in chloroform and washed twice with a saturated aqueous sodium hydrogen carbonate solution and twice with water. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: chloroform-methanol, 40: 1) to give the title compound (4.00 g, 86%) as a colorless foam.

[Α] ²⁴ _D + 12.8 ° (c 1.09, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.34 (3H,
s, isopropylidene), 1.44 (12H, s, COOC (CH 3) 3, isopropylidene), 1.94 (1H, t, J = 5.9 Hz, OH), 2.04 (1
H, m, 5-H), 3.17 (1H, t, J = 12.2 Hz, 6-Hax), 3.45 (1
H, dd, J = 12.2,4.4 Hz, 6-Heq), 3.75 (2H, t, J = 5.9 H
z, -C H ₂ OH), 4.53 (1H, dd, J = 7.8, 2.4Hz, 3-H), 4.57
(1H, broad d, J = 7.8 Hz, 4-H), 4.87 (1H, broad, -NHC
O-), 5.08, 5.13 (1H, ABq, J = 12.0Hz, PhC H ₂ O-), 5.65 (1
H, dd, J = 6.1, 2.4Hz, 2-H), 7.25 (1H, s, Ph ₂ C H =), 7.2
9-7.35 (5.4, m, Ph).

(2) (3S, 4S, 5R, 6S) -6-
Benzyloxycarbonylamino-1-N- (tert-butoxycarbonyl) -4,5-dihydroxy-4,5-O-
Isopropylidenepiperidine-3-carboxylic acid [formula (XI-
1) Compound] The compound of the formula (X-1) (2.18 g) obtained in the above item (1) is mixed with carbon tetrachloride-acetonitrile (1: 1) mixed solvent (6).
0 ml) and to this solution was added a solution of sodium metaperiodate (3.12 g) in water (45 ml). Ruthenium (IV) dioxide (8.
0 mg) was added, and the mixture was vigorously stirred at room temperature for 40 minutes to carry out an oxidation reaction. The precipitated off-white solid was removed by filtration under reduced pressure, and the organic phase and aqueous phase were separated.

The aqueous phase was extracted twice with ethyl acetate. The ethyl acetate extract and the organic phase are combined and combined with 2-propanol (5 m
l) was added and the mixture was stirred at room temperature for 2 hours. After removing the precipitated solid using Kiriyama wax, the filtrate was washed once with water.
The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was subjected to silica gel column chromatography (elution solvent: chloroform-
Purify with methanol-concentrated aqueous ammonia, 60: 10: 1) to give the title compound as a colorless amorphous solid (1.79 g, 90).
%).

[Α] ²³ _D + 5.7 ° (c 0.96, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.32, 1.
37 (3H, s, isopropylidene), 1.44 (9H, s, COOC (CH ₃ )
₃ ), 2.96 (1H, ddd, J = 12.7, 5.4, 2.9 Hz, 3-H), 3.46 (1
H, t, J = 12.7 Hz, 2-Hax), 3.55 (1H, dd, J = 12.7, 5.4
Hz, 2-Heq), 4.45 (1H, dd, J = 7.6, 2.0 Hz, 5-H), 4.82
(1H, dd, J = 7.6, 2.9 Hz, 4-H), 5.08,5.12 (1H, ABq, J
= 12.7 Hz, PhC H ₂ O-), 5.63 (1H, d, J = 2.0 Hz, 6-H), 7.
27-7.38 (5H, m, Ph).

(3) (3S, 4S, 5R, 6S) -6-
Benzyloxycarbonylamino-1-N- (tert-butoxycarbonyl) -4,5-dihydroxy-4,5-O-
Preparation of isopropylidenepiperidine-3-carboxylic acid diphenylmethyl ester [compound of formula (XII-1)] The compound (117 mg) of formula (XI-1) obtained in the above (2) was diluted with dichloromethane (1.5 ml) -methanol. This was dissolved in a mixed solvent (1.5 ml), diphenyldiazomethane (0.152 g) was added to the solution, and the mixture was stirred overnight at room temperature to carry out an esterification reaction. Acetic acid (60 μl) was added to the reaction solution to stop the reaction, the mixture was stirred at room temperature for 30 minutes, and the solvent was evaporated under reduced pressure. The residue was dissolved in chloroform and the solution was washed once with saturated aqueous sodium hydrogen carbonate solution and three times with water. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: hexane-ethyl acetate, 3: 1) to give the title compound (150 mg, 94%) as a colorless foam.

[Α] ²⁴ _D + 3.55 ° (c 0.87, MeOH): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.33, 1.
36 (3H, s, isopropylidene), 1.43 (9H, s, COOC (CH ₃ )
₃ ), 3.49 (1H, t, J = 12.7 Hz, 2-Hax), 3.64 (1H, dd, J = 1
2.2, 4.9 Hz, 2-Heq), 4.52 (1H, dd, J = 7.8, 2.0 Hz, 5
-H), 4.93 (1H, dd, 7.8, 2.4 Hz, 4-H), 5.07, 5.13 (1H,
ABq, J = 12.2 Hz, PhC H ₂ O-), 5.66 (1H, d, J = 2.0 Hz, 6-
H), 6.89 (1H, s, Ph ₂ C H O-), 7.25-7.37 (15H, m, Ph).

(4) Diphenylmethyl (5S, 6S) -6-benzyloxycarbonylamino-1-N- (tert-butoxycarbonyl) -3,4-didehydropiperidine-3-carboxylate [formula (XIII-1 Compound of formula (XII-1) (799 mg) under an argon atmosphere.
Was dissolved in tetrahydrofuran and the solution was cooled to 0 ° C. Tertiary butoxy potassium (15 mg)
Was added and the mixture was stirred at 0 ° C. for 2 hours to carry out the reaction of deoxygenation and enolization. The reaction solution was diluted with chloroform, washed twice with a saturated aqueous solution of ammonium chloride, twice with water, and once with a small amount of a saturated aqueous solution of sodium chloride. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: toluene-acetone, 7: 1) to give the title compound (508 mg, 70%) as a colorless foam.

[Α] ²⁴ _D + 62.9 ° (c 0.89, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.47 (9
H, s, COOC (CH ₃ ) ₃ ), 3.81 (1H, d, J = 19.0 Hz, 2-Ha),
4.19 (1H, d, J = 5.4 Hz, 5-H), 4.53 (1H, d, J = 19.0Hz,
2-Hb), 5.06, 5.10 (1H, ABq, J = 12.7 Hz, PhC H ₂ O-), 5.
92 (1H, d, J = 1 Hz, 6-H), 6.95 (1H, s, Ph ₂ C H O-), 7.70
(1H, m, 4-H), 7.26-7.40 (1.5H, m, Ph).

(5) (3aS, 4R, 7S, 7aS) -7
-(Benzyloxycarbonylamino) -6-N- (tert-butoxycarbonyl) -2- (trichloromethyl) -4-
(Diphenylmethyloxycarbonyl) -3a, 7a-dihydrooxazolo [5,4-c] piperidine [Formula (XIV-1)
Compound of formula (XIII-1) (168 mg) obtained in the previous section (4) was dissolved in benzene (4 ml), and trichloroacetonitrile (39 μl) and DBU (9.0 μl) were added to the solution,
The reaction was carried out by stirring at room temperature for 45 minutes. The reaction was stopped by adding saturated ammonium chloride aqueous solution to the reaction solution. The reaction solution was diluted with chloroform and washed with water three times. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: toluene-acetone, 50: 1) to give the title compound as a colorless foam (14
7 mg, 70%) was obtained.

[Α] ²⁶ _D + 2.0 ° (c 0.89, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.42 (9H,
s, COOC (CH ₃ ) ₃ ), 3.39 (1H, broad t, J = 11.7 Hz, 2-Ha
x), 3.54 (1H, ddd, J = 12.7, 4.9, 4.4 Hz, 3-H), 3.64 (1
H, dd, J = 11.7, 4.9 Hz, 2-Heq), 5.09, 5.14 (1H, ABq,
J = 11.7 Hz, PhC H ₂ O-), 5.18 (1H, dd, J = 9.8, 4.4 Hz, 4
-H), 5.27 (1H, broad d, J = 9.8 Hz, 5-H), 5.82 (1H, d,
J = 2.0 Hz, 6-H), 6.92 (1H, s, Ph ₂ C H O-), 7.23-7.41 (15
H, m, Ph).

(6) (3R, 4S, 5S, 6S) -6-Benzyloxycarbonylamino-1-N- (tert-butoxycarbonyl) -5-hydroxy-4-trichloroacetamidepiperidine-3-carboxylic acid diphenylmethyl Production of ester [compound of formula (XV-1)] The compound of formula (XIV-1) (1.55 g) obtained in the preceding paragraph (5) was dissolved in a mixed solvent of pyridine (24 ml) -water (6 ml),
Para-toluenesulfonic acid (monohydrate, 63
8 mg) was added and the mixture was stirred at 80 ° C. for 3 hours. After the obtained reaction solution was cooled to room temperature, the solvent was distilled off under reduced pressure. The residue was dissolved in chloroform and washed twice with saturated aqueous sodium hydrogen carbonate solution and twice with water. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was azeotropically distilled with toluene three times, and then purified by silica gel column chromatography (eluting solvent: chloroform-methanol, 30: 1) to obtain the title compound as colorless foam (1.51 g, 95%).

[Α] ²⁶ _D -5.7 ° (c 0.92, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.45 (9
H, s, COOC (CH ₃ ) ₃ ), 3.14 (1H, broad t, J = 12.4 Hz, 2-
Hax), 3.23 (1H, dt, J = 11.7, 4.1 Hz, 3-H), 3.84 (1H,
broad s, 5-H), 4.24 (1H, broad d, J = 12.2 Hz, 2-He
q), 4.52 (1H, dd, J = 11.5, 2.7 Hz, 4-H), 5.10, 5.14 (1
H, ABq, J = 12.7 Hz, PhC H ₂ O-), 5.87 (1H, d, J = 2.0 Hz,
6-H), 6.83 (1H, s, Ph ₂ C H O-), 7.24-7.38 (15H, m, P
h).

(7) (3R, 4S, 5S, 6S) -6-
Benzyloxycarbonylamino-4-amino-1-N- (tert-butoxycarbonyl) -5-hydroxypiperidine-3-carboxylic acid diphenylmethyl ester [Formula (XV
I-1) compound] The compound of formula (XV-1) (144 mg) was added to ethanol (3 ml).
, Sodium borohydride (23 mg) was added thereto, and the mixture was stirred at room temperature for 35 minutes. Removal of the trichloroacetyl group was performed. The obtained reaction solution was diluted with chloroform and washed twice with a saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: chloroform-methanol, 20: 1) to give the title compound (57 mg) as a colorless foam. The portion that could not be purified completely was re-purified by preparative TLC (chloroform / methanol, 15: 1) to obtain the title compound (21 mg) as a colorless amorphous solid.

[Α] ²⁶ _D + 6.3 ° (c 1.90, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.44 (9
H, s, COOC (CH ₃ ) ₃ ), 2.84 (1H, dt, J = 11.4, 4.1 Hz, 3-
H), 2.99 (1H, t, J = 12.5 Hz, 2-Hax), 3.22 (1H, dd, J = 1
0.7, 2.9 Hz, 4-H), 3.74 (1H, broad s, 5-H), 4.22 (1
H, dd, J = 13.4,4.6 Hz, 2-Heq), 5.09 (2H, s, PhC H ₂ O
-), 5.86 (1H, d, J = 1.5 Hz, 6-H), 6.90 (1H, s, Ph ₂ C H O
-), 7.25-7.40 (15H, m, Ph).

(8) (3R, 4S, 5S, 6S) -6-
Benzyloxycarbonylamino-1-N- (tert-butoxycarbonylamino) -4- (tert-butoxycarbonyl) -5-hydroxypiperidine-3-carboxylic acid diphenylmethyl ester [compound of formula (XVII-1)] The compound of formula (XVI-1) (81 mg) was added to methanol (1.6 m
l) and N, N-diisopropylethylamine (73.2 μl) and di-tert-butyl dicarbonate (64.3 μl) were added to the solution. The reaction for protecting the amino group was carried out by stirring at room temperature for 1 hour. Then, the solvent was distilled off from the reaction solution under reduced pressure. The residue was dissolved in chloroform and washed twice with 10% aqueous citric acid solution. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The filtrate was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: toluene-acetone, 8: 1) to give the title compound as a colorless amorphous solid (87 mg, 91).
%).

[Α] ²⁶ _D -3.1 ° (c 0.93, MeOH): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.33, 1.
42 (9H, s, COOC (CH ₃ ) ₃ ), 2.96 (1H, dt, J = 11.7, 4.1 Hz,
3-H), 3.11 (1H, t, J = 12.7 Hz, 2Hax), 3.77 (1H, m, 5
-H), 4.12 (2H, m, 2-Heq, 4-H), 5.08, 5.11 (1H, ABq,
J = 12.5 Hz, PhC H ₂ O-), 5.83 (1H, d, J = ~ 2.0 Hz, 6-H),
6.81 (1H, s, Ph ₂ C H O-), 7.23-7.37 (15H, m, Ph).

(9) (3R, 4S, 5S, 6S) -6-
Benzyloxycarbonylamino-1-N- (tert-butoxycarbonyl) -4- (tert-butoxycarbonylamino) -5-hydroxypiperidine-3-carboxylic acid (2-methoxy) ethoxymethyl ester [Formula (XVIII-
Compound of formula (XVII-1) (20 mg) was added to methanol (0.4 m)
l) and dissolved in 1M aqueous sodium hydroxide solution (0.
15 ml) was added. Alkaline hydrolysis was carried out by stirring at room temperature for 1.5 hours. The solvent was distilled off under reduced pressure from the reaction solution. The colorless solid obtained contains a compound of formula (XVII-a-1). Chloroform and a 10% citric acid aqueous solution were added thereto, and the mixture was stirred and then separated into an organic phase and an aqueous phase. The aqueous phase was extracted twice with chloroform. The chloroform extract was combined with the organic phase, dried over anhydrous magnesium sulfate, and filtered through a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was dissolved in dichloromethane (0.4 ml) and the solution cooled to 0 ° C. Add N, N-diisopropylethylamine (6.3 μm
1) and chloromethyl (2-methoxy) ethyl ether (4.1 μl) were added, and the mixture was stirred at 0 ° C. for 1.5 hours to carry out a reesterification reaction. The reaction solution was diluted with chloroform and washed with water three times. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was collected by preparative TLC (toluene / acetone, 3:
1) and the title compound as a colorless solid (15 mg, 83
%).

[Α] ²⁵ _D -2.4 ° (c 0.67, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.42, 1.
44 (9H, s, COOC (CH ₃ ) ₃ ), 2.84 (1H, dt, J = 11.7, 4.4 Hz,
3-H), 3.15 (1H, broad t, J = 13.2 Hz, 2-Hax), 3.36 (3
H, s, -OCH ₃ ), 3.55, 3.79 (2H, m, -OCH ₂ CH ₂ O), 3.77 (1
H, m, 5-H), 4.06 (1H, dd, J = 11.2, 2.5 Hz, 4-H), 4.14
(1H, J = 13.2, 3.7 Hz, 2-Hap), 5.10, 5.13 (1H, ABq, J =
12.5 Hz, PhC H ₂ O-), 5.28, 5.31 (1H, ABq, J = 6.1 Hz, -C
O ₂ CH ₂ O-), 5.83 (1H, d, 6-H), 7.27-7.39 (5H, m, Ph).

(10) (3R, 4S, 5S, 6S) -N-
(Tert-butoxycarbonyl) -4- (tert-butoxycarbonylamino) -5-hydroxy-6- (trifluoroacetamido) piperidine-3-carboxylic acid (2-
Preparation of methoxy) ethoxymethyl ester [compound of formula (XIX-1)] A compound of formula (XVIII-1) (43 mg) was dissolved in acetonitrile (2 ml), and 10% palladium carbon (15
mg), and stirred under a hydrogen atmosphere at room temperature for 30 minutes,
The elimination reaction of the benzyloxycarboxyl group was performed. 1
After 0% palladium carbon was filtered off under reduced pressure, the solvent of the filtrate was distilled off under reduced pressure. A residue containing a compound of formula (XVIII-a-1) was obtained.

The residue was dissolved in dichloromethane (0.8 ml) and the solution cooled to 0 ° C. Pyridine (13
μl) and trifluoroacetic anhydride (11 μl) were added to 0
A trifluoroacetylation reaction was performed by stirring at 20 ° C. for 20 minutes. Pyridine (13 μl) and trifluoroacetic anhydride (11 μl) were added to the reaction solution, and the mixture was further added at 0 ° C. for 20 hours.
Stirred for minutes. The reaction solution was diluted with chloroform and washed once with water, twice with a saturated aqueous sodium hydrogen carbonate solution, and twice with water. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was azeotropically distilled three times with toluene and then purified by preparative TLC (developing solvent: chloroform-methanol, 25: 1),
The title compound (27 mg, 68%) was obtained as a colorless solid.

[Α] ²⁷ _D + 8.8 ° (c 1.08, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.43, 1.
46 (9H, s, COOC (CH ₃ ) ₃ ), 2.92 (1H, dt, J = 11.7, 4.0 Hz,
3-H), 3.27 (1H, broad t, J = 11.7 Hz, 2-Hax), 3.37 (3
H, s, -OCH ₃ ), 3.55 (2H, m, -OC H ₂ CH ₂ O-), 3.81 (3H, m,
-OCH ₂ C H ₂ O-, 5-H), 4.04 (1H, broad d, J = 10.7 Hz, 4-
H), 4.21 (1H, dd, J = 13.7, 3.9 Hz, 2-Heq), 5.30, 5.33
(1H, ABq, J = 6.1 Hz, -CO ₂ C H ₂ O-), 6.01 (1H, s, 6-H).

(11) (3R, 4S, 5S, 6R) -4-
Production of Amino-5-hydroxy-6-trifluoroacetamidopiperidine-3-carboxylic acid [compound of formula (Ic)] Compound of formula (XIX-1) (22 mg) was added to 4M hydrochloric acid-dioxane solution (0.7 ml). After dissolution, the solution was stirred at room temperature overnight to carry out a deprotection reaction. After adding diethyl ether to the reaction solution in which a colorless solid has precipitated and is suspended and stirred well,
The deposited colorless solid was precipitated by centrifugation and the supernatant was removed. The obtained precipitate was air-dried and then dried under reduced pressure with a pump to obtain the title compound as a colorless solid, that is, the dihydrochloride of the present compound No. 3 (15 mg, 112%).

Example 4 (1) (3R, 4S, 5S, 6S) -6-Benzyloxycarbonylamino-1-N- (tertiary butoxycarbonyl) -4- [N, N'-di- (tertiary Butoxycarbonyl)] Guanidino-5-hydroxypiperidine-3-carboxylic acid diphenylmethyl ester [compound of formula (XX-1)] The compound of formula (XVI-1) (0.1) obtained in Example 3 (7). 207
g) was dissolved in N, N-dimethylformamide (4 ml) and the solution was cooled to 0 ° C. Triethylamine (0.20 ml), N, N'-di- (tert-butoxycarbonyl) thiourea (0.199 g), mercuric chloride (I
I) (0.196 g) was added, and the mixture was stirred at 0 ° C. for 1.5 hours to carry out an amidinoization reaction. Ethyl acetate was added to the reaction solution in which an ocher-colored solid had precipitated, and the solid was precipitated by centrifugation, and the supernatant was transferred separately. After that, suspend the precipitate in ethyl acetate, centrifuge, and then transfer the supernatant separately.
Repeated times. The supernatants were combined and washed 3 times with water and once with a small amount of saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue was azeotropically distilled with toluene three times and then subjected to silica gel column chromatography (developing solvent:
Purification with toluene-acetone, 10: 1) gave the title compound as a colorless foam (0.29 g, 98%).

[Α] ²⁵ _D -7.7 ° (c 0.98, CHCl ₃ ): ¹ H-NMR spectrum (CD ₃ OD, 40 ° C., δppm): 1.46,
1.47, 1.48 (9H, s, COOC (CH ₃ ) ₃ ), 2.95 (1H, dt, J = 11.7,
3.9 Hz, 3-H), 3.13 (1H, broad t, J = 12.7Hz, 2-Hax,),
3.74 (1H, broad s, 5-H), 4.14 (1H, broad d, J = 12.7
Hz, 2-Heq), 5.11 (2H, broad s, PhC H ₂ O-), 5.85 (1H, bro
ad s, 6-H), 6.80 (1H, s, Ph ₂ C H O-), 7.24-7.38 (15H, m,
Ph).

(2) (3R, 4S, 5S, 6S) -1-N
-(Tertiary butoxycarbonyl) -4- [N, N'-di-
(Tertiary butoxycarbonyl)] guanidino-5-
Hydroxy-6-trifluoroacetamidopiperidone-
3-carboxylic acid diphenylmethyl ester [Formula (XXI-1)
Compound of formula (XX-1) (188 mg)
l) and add 10% palladium on carbon (63m
g) was added, and the mixture was stirred under a hydrogen atmosphere at room temperature for 8 hours to carry out the elimination reaction of the benzyloxycarbonyl group. After 10% palladium carbon was filtered off under reduced pressure, the solvent of the filtrate was distilled off under reduced pressure. The residue obtained contains the compound of formula (XX-a-1).

The residue was dissolved in dichloromethane (4 ml) and cooled to 0 ° C. Pyridine (40 μl) and trifluoroacetic anhydride (35 μl) were added thereto, and the mixture was stirred at 0 ° C. for 30 minutes to carry out a trifluoroacetylation reaction. Pyridine (40 μl) and trifluoroacetic anhydride were added to the reaction solution.
(35 μl) was added, and the mixture was further stirred at 0 ° C. for 30 minutes.
The reaction solution was diluted with chloroform and washed once with water, twice with a saturated aqueous sodium hydrogen carbonate solution, and twice with water. The organic phase was dried over anhydrous magnesium sulfate and then filtered with a cotton plug. The solvent of the filtrate was distilled off under reduced pressure. The residue is 3 with toluene
After azeotropic distillation twice, the residue was purified by preparative TLC (developing solvent: toluene-acetone, 8: 1) to give the title compound (3
8 mg, 33%) was obtained. At this time, unreacted compound of formula (XX-1) (68 mg, 36%) was recovered.

[Α] ²⁴ _D -6.7 ° (c 0.78, CHCl ₃ ): ¹ H-NMR spectrum (CD ³ OD, 40 ° C., δppm): 1.471,
1.477, 1.480 (9H, s, COOC (CH ₃ ) ₃ ), 3.01 (1H, dt, J = 11.
7, 4.4 Hz, 3-H), 3.19 (1H, broad t, J = 12.2Hz, 2-Ha
x), 3.84 (1H, broad s, 5-H), 4.21 (1H, broad d, J = 1
2.6 Hz, 2-Heq), 6.04 (1H, broad d, J = 1.5 Hz, 6-H),
6.82 (1H, s, Ph ₂ C H O-), 7.26-7.34 (10H, m, Ph).

(3) (3R, 4S, 5S, 6R) -5-
Preparation of hydroxy-4-guanidino-6-trifluoroacetamidopiperidine-3-carboxylic acid [compound of formula (Id)] Compound of formula (XXI-1) (44 mg) in 4M hydrochloric acid-dioxane solution (1.3 ml). After dissolution, the solution was stirred at room temperature overnight to carry out a deprotection reaction. After adding diethyl ether to the reaction solution in which a colorless solid has precipitated and is suspended and stirred well,
The deposited colorless solid was precipitated by centrifugation and the supernatant was removed. The obtained precipitate was air-dried and then dried under reduced pressure by a pump to obtain the title compound as a colorless solid, that is, the dihydrochloride salt of compound No. 4 of the present invention (21 mg, 100%).

Reference Example 1 (a) N- (tert-butoxycarbonyl) -4,5-O
-Production of isopropylidene cystatin B N- (tert-butoxycarbonyl) -cystatin B
(0.955 g, 3 mmol) was suspended in N, N-dimethylformamide (DMF) (15 ml), to which 2,2-dimethoxypropane (3.69 ml, 30 mmol) and chlorotrimethylsilane (1.90 ml, 15 mmol) were suspended. ) Was added.
After stirring the resulting mixture at room temperature for 2 hours, pyridine (1.5 ml) was added to the reaction solution to stop the reaction.

The reaction solution was diluted with chloroform, water was added thereto for extraction, and then the aqueous phase was extracted three times with a mixed solution of chloroform-methanol (9: 1). The organic phases were combined, dried over anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and the residue was azeotropically distilled three times with toluene,
It was purified by silica gel column chromatography using chloroform-methanol-concentrated aqueous ammonia (2: 1: 0.3) as a developing solvent. The following formula is given as a colorless foamy solid. 1.05 g (98%) of the title compound shown by the formula (Boc and Me have the same meanings as described above) was obtained. Specific rotation: [α] ²³ _D + 29 ° (c 0.88, methanol).

(B) 2-of N- (tert-butoxycarbonyl) -4, -5-O-isopropylideneciastatin B
Preparation of methoxyethoxymethyl ester Compound of formula (C ') obtained in (a) above (2.51 g, 7 mmo
l) was dissolved in DMF (50 ml) and to this was added diisopropylethylamine (4.88 ml, 28 mmol) and 2-methoxyethoxymethyl chloride (1.60 ml, 14 mmol). The resulting mixture was stirred at room temperature for 2 hours. afterwards,
The reaction was stopped by adding water (0.2 ml) to the reaction solution.

The reaction solution was diluted with chloroform, water was added thereto for extraction, and then the aqueous phase was extracted three times with chloroform. The organic phases were combined, dried over anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, the residue was azeotropically distilled three times with toluene, and then purified by silica gel column chromatography using chloromethane-methanol (20: 1) as a developing solvent. The following formula for colorless syrup 2.61 g (83%) of the title compound represented by the formula (Boc, Me and MEM have the same meaning as described above) was obtained.
Specific rotation: [α] ²⁵ _D -18.7 ° (c 0.91, chloroform).

(C) (3R, 4S, 5R, 6S) -6-
Acetamide-N- (tertiary butoxycarbonyl) -3-
Preparation of Hydroxymethyl-4,5-dihydroxy-4,5-O-isopropylidenepiperidine Compound of formula (D ') obtained in the above (b) (2.59 g, 5.8)
mmol) was dissolved in a mixed solvent of tetrahydrofuran (50 ml) and 2,2,2-trifluoroethanol (5 ml), and sodium borohydride (0.685 g, 17.4 mm) was added to the solution.
ol). The obtained mixture was stirred at room temperature for 1 hour to carry out a reduction reaction. After that, water (1.5 m) was added to the reaction solution.
l) was added to stop the reaction, and the mixture was further stirred at room temperature for 30 minutes. The obtained reaction solution was concentrated under reduced pressure, water was added to the residue, and the mixture was extracted 3 times with chloroform. The organic phases (chloroform extract) were combined, dried over anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and the residue was used as a developing solvent in dichloromethane-methanol (12: 1).
Purified by silica gel column chromatography using 1.98 g (99%) of the title compound was obtained. Specific rotation:
[α] 25D + 27 ° (c 0.95, chloroform).

(D) (3R, 4S, 5R, 6S) -6-
Production of amino-N- (tert-butoxycarbonyl) -3-hydroxymethyl-4,5-dihydroxy-4,5-O-isopropylidenepiperidine Compound of formula (E ') obtained in the above (c) (1. 03g, 3mm
ol) to hydrazine hydrate (H ₂ NNH ₂ · xH ₂ O, 20 m
l) and stirred at 70 ° C. for 10 days. Formula (E ')
A reaction occurred in which the acetyl group was eliminated from the 6-position acetamide group of the compound (1). After cooling the reaction solution to room temperature, the reaction solution was concentrated under reduced pressure. Water was added to the residue, and the mixture was extracted 3 times with chloroform. Organic phase (chloroform extract)
Were combined, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography using ethyl acetate-methanol (50: 1) as a developing solvent and ethyl acetate as a developing solvent.
-Silica gel preparative T with methanol (10: 1)
Purified by LC chromatography. The following formula for colorless syrup 0.488 g (54%) of the title compound shown by was obtained. Specific rotation: [α] ²⁶ _D -14.4 ° (c 0.99, chloroform). Further, at this time, the unreacted compound of the formula (E ') is reduced to 0.
332 g (32%) was recovered.

When the compound of the formula (F ') above is dissolved in 4M hydrochloric acid-dioxane and the solution is stirred overnight at room temperature to carry out the deprotection reaction, the compound represented by the above formula (A) ( Three
R, 4S, 5R, 6S) -6-Amino-3-hydroxymethyl-4,5-dihydroxypiperidine was obtained as a colorless substance.

Claims (9)

[Claims] 1. Formula (I) (In the formula, R ¹ is an acetyl group or a trifluoroacetyl group, and R ² is an amino group or a guanidino group), and a 3-epicistatin B derivative and a salt thereof. 2. R ^{1 in the} general formula (I) shown in claim ^1.
Is an acetyl group and R ² is an amino group, that is, the following formula (Ia) The compound according to claim 1, which is (3R, 4S, 5S, 6R) -6-acetamido-4-amino-5-hydroxypiperidine-3-carboxylic acid or a salt thereof. 3. In the general formula (I) shown in claim 1, R ¹
Is an acetyl group and R ² is a guanidino group, that is, the following formula (Ib) (3R, 4S, 5S, 6R) -6-acetamide
The compound according to claim 1, which is -4-guanidino-5-hydroxypiperidine-3-carboxylic acid or a salt thereof. 4. R ^{1 in the} general formula (I) shown in claim ^1.
Is a trifluoroacetyl group and R ² is an amino group, that is, the following formula (Ic) (3R, 4S, 5S, 6R) -4-amino-5-hydroxy-6-trifluoroacetamidopiperidine-3-
The compound according to claim 1, which is a carboxylic acid or a salt thereof. 5. R ^{1 in the} general formula (I) shown in claim ^1.
Is a trifluoroacetyl group and R ² is a guanidino group, that is, the following formula (Id) (3R, 4S, 5S, 6R) -4-guanidino-
The compound according to claim 1, which is 5-hydroxy-6-trifluoroacetamidopiperidine-3-carboxylic acid or a salt thereof. 6. The following formula (II) The following formula (III) derived from cystatin B represented by (In the formula, R represents an imino protecting group), N-protected-
The carboxyl group of 4-deoxy-3-enciastatin B is reacted with an esterifying agent to esterify it, and the compound of the following formula (IV) (Wherein R has the same meaning as described above and R ^a represents a carboxyl-protecting group that is an ester-forming group), an ester compound is formed, and then the compound of formula (IV) is reacted with trichloroacetonitrile. Thereby forming an oxazoline ring, (Wherein R and R ^a have the same meanings as described above), and then the compound of formula (V) is hydrolyzed to open the oxazoline ring, whereby the following formula (VI) (Wherein, R and R ^a have the same meanings as defined above) to produce a compound represented by the further trichloroacetyl group elimination from the compound of formula (VI) the following formula (VII) (Wherein R and R ^a have the same meanings as described above), and from the compound of formula (VII) the imino protecting group (R) and the carboxyl protecting group (R ^a )
The following equation (Ia) characterized by desorbing (3R, 4S, 5S, 6R) -6-acetamide
-4-amino-5-hydroxypiperidine-3-carboxylic acid,
Or the manufacturing method of the salt. 7. The following formula (VII) (Wherein R represents an imino protecting group and R ^a represents a carboxyl protecting group which is an ester forming group) (3
R, 4S, 5S, 6R) -6-acetamido-4-amino-5
-Hydroxypiperidine-3-carboxylic acid protected derivative (In the formula, A represents an amino-protecting group which is an alkoxycarbonyl group, an aryloxycarbonyl group or an aralkyloxycarbonyl group), and N, N'-di-protected-
By reacting with thiourea, it is converted to amidino and is converted to the following formula (VIII) (Wherein R, ^Ra and A have the same meanings as described above), and then the respective protecting groups (R, ^Ra and A) are eliminated from the compound of formula (VIII). Which is characterized by the following equation (Ib) (3R, 4S, 5S, 6R) -6-acetamide
A method for producing 4-guanidino-5-hydroxypiperidine-3-carboxylic acid and a salt thereof. 8. The following formula (IX) derived from cystatin B by a known method: (Wherein R represents an imino protecting group, X ¹ and X ² are each a monovalent hydroxyl protecting group, or both represent a divalent hydroxyl protecting group together) (3R , 4S, 5R, 6S) -N-Protected-6-amino-3
-Hydroxymethyl-4,5-dihydroxy-4,5-O-di
-Protection-The 6-position amino group of piperidine is replaced with an imino protecting group (R)
Protected with an amino protecting group (R ^b ) which can be eliminated by a different elimination method from (Wherein R, R ^b , X ¹ and X ² have the same meanings as described above), and then the 3-hydroxymethyl group of the compound of formula (X) is oxidized to a carboxyl group. Convert to the following formula (XI) (Wherein R and R ^b and X ¹ and X ² have the same meanings as defined above) to produce a carboxylic acid compound, and then reacting the compound of formula (XI) with an esterifying agent (XII) (In the formula, R, X ¹ and X ² have the same meanings as described above, R ^a represents a carboxyl-protecting group which is an ester-forming group, and R ^b represents an amino-protecting group which can be eliminated by an elimination method different from R. Represent)
A compound of formula (XII) is removed (deoxygenation) of the 4-position hydroxyl group and the bond between the 3- and 4-position carbons is converted to a double bond. Formula (XIII) (Wherein R, R ^a and R ^b have the same meanings as described above), and then the compound of formula (XIII) is reacted with trichloroacetonitrile to form an oxazoline ring. Formula (XIV) (Wherein R, R ^a and R ^b have the same meanings as described above), and then the compound of formula (XIV) is hydrolyzed to open the oxazoline ring of the compound of formula (XIV). Then, the following formula (XV) (Wherein R, R ^a and R ^b have the same meanings as described above), and the trichloroacetyl group is eliminated from the compound of formula (XV) to give the following formula (XVI) (Wherein R, R ^a, and R ^b have the same meanings as described above), and the 4-position amino group of the compound of formula (XVI) is protected with the same amino group as the imino protecting group (R). Protected by a group of formula (XVII) (Wherein R, R ^a and R ^b have the same meanings as described above), and the carboxyl protecting group (R ^a ) is eliminated from the compound of formula (XVII) to give the following formula (XVII- a) A method of producing a compound represented by the formula (wherein R and R ^b have the same meanings as described above) and removing the imino protecting group (R) from the 3-position carboxyl group of the compound of formula (XVII-a) Protected by a carboxyl protecting group (R ^c ) which can be eliminated in the same reaction as in the following formula (XVIII) (Wherein, R and R ^b have the same meanings as defined above, R ^c is a carboxyl protecting group) to produce a compound represented by the following amino protecting group from the compound of formula (XVIII) and (R ^b) The following formula (XVIII-a) (Wherein R and R ^c have the same meanings as described above), and the amino group at the 6-position of the compound of formula (XVIII-a) is trifluoroacetylated to give the following formula (XIX) (Wherein R and R ^c have the same meaning as described above), and then the amino protecting group (R) and the carboxyl protecting group (R ^c ) are removed from the compound of formula (XIX). The following equation (Ic) characterized by (3R, 4S, 5S, 6R) -4-amino-5-hydroxy-6-trifluoroacetamidopiperidine-3-
Process for producing carboxylic acid and its salt. 9. The following formula (XVI) [In the formula, R represents an imino protecting group, R ^a represents a carboxyl protecting group which is an ester forming group, and R ^b represents an amino protecting group which can be eliminated by a different elimination method from the imino protecting group (R)]. Indicated by (3R, 4S, 5S, 6R or 6
The protected derivative of (S) -6-N-protected-4,6-diamino-5-hydroxypiperidine-3-carboxylic acid is represented by the following formula (Wherein A represents an amino-protecting group which is an alkoxycarbonyl group, an aryloxycarbonyl group or an aralkyloxycarbonyl group), and is N, N'-di-protected.
-The compound of the following formula (XX) (Wherein R, R ^a , R ^b and A have the same meanings as described above), and then the amino protecting group (R ^b ) at the 6-position of the compound of formula (XX) is selectively formed. To the next equation
(XX-a) (Wherein R, ^Ra and A have the same meanings as described above), and the amino group at the 6-position of the compound of formula (XX-a) is trifluoroacetylated to give the following formula (XXI) (Wherein R, R ^a and A have the same meanings as described above), and the respective protecting groups (R, R ^a and A) are eliminated from the compound of formula (XXI). Which is characterized by the following expression (Id) (3R, 4S, 5S, 6R) -4-guanidino-
Process for producing 5-hydroxy-6-trifluoroacetamidopiperidine-3-carboxylic acid, and salts thereof.