JPH0725823A - Water-soluble benastatin derivative and pharmaceutical use thereof - Google Patents

Abstract

PURPOSE:To obtain a new benastatin derivative, having strong immun opotentiating activity at a low concentration and immunosuppressive activity at a high concentration and useful as an immunomodulator and a physiological ly permissible salt thereof. CONSTITUTION:This benastatin derivative is expressed by formula I {R<1> is CH=CH or CH2CH2; R<2> is CO-(CH2)n-Y or (CH2)n+1-Y [n is 0-5: Y is CO2H or NR<3>R<4> (R<3> and R<4> are H or 1-5C alkyl or R<3> and R<4> are bound to represent a lower alkylene which may contain O or N in the binding chain thereof)]} and a physiologically permissible salt thereof, e.g. 1-(4-carboxybutyryl)benastatin C. The compound expressed by formula ], e.g. the benastatin derivative in which R<2> is CO-(CH2)n-CO2H and (n) is 2-5 is obtained by reacting the benastatin C or D with a dicarboxylic acid anhydride expressed by formula II by using a base such as potassium carbonate or pyridine in a solvent such as hexane or benzene.

Description

Detailed Description of the Invention

[0001]

The present invention relates to an immunomodulator (immun
The present invention relates to a novel water-soluble Benastatin derivative useful as an omodulator) and its pharmaceutical use.

[0002]

2. Description of the Related Art Immunosuppressants such as cyclosporin A and immunostimulants such as Krestin (registered trademark) and Ubenimex (generic name) are known as immunomodulators.

[0003]

Benastatin C and D used as raw materials in the present invention are Streptomyces sp. MI384-DF12 isolated by Takaaki Aoyagi et al.
It is a compound that was isolated from a fermentation product of a strain (Microtechnology Research Institute No. 3228) as a novel physiologically active substance and found to have a strong immunostimulatory action and an immunosuppressive action at a high concentration. The chemical structures of Benastatin C and D are as follows:

[Chemical 2] Benasutachin C: R ¹ = -CH = CH- Benasutachin _{^{D: R 1 = -CH 2 CH}} 2 - Benasutachin C and D but is expected as adjuvants, these compounds have a problem in terms of water-soluble, For example, Benastatin C is poorly soluble in water, and thus it was considered necessary to devise a formulation when formulating it. Therefore, development of Benastatin C and D derivatives which are excellent in water solubility is desired.

[0004]

Therefore, as a result of various studies, the present inventors have introduced a specific substituent into the hydroxyl group at the 1-position present in the molecules of benastatin C and D,
It was found that a benastatin derivative having significantly improved solubility in water was obtained, and that this derivative has a strong immunostimulatory activity at a low concentration and an immunosuppressive activity at a high concentration, and thus completed the present invention. Therefore, the present invention provides the following formula (1)

[Chemical 3] Wherein, R ¹ is -CH = CH- or -CH ₂ CH ₂ - indicates, R ² is -CO- (CH _{2) n} -Y or - (C
Shows the H _{2) n +} 1 -Y. Where n is an an integer of 0 to 5, Y is -CO ₂ H or NR ³ R ⁴ (R ^3, R ⁴ are each a hydrogen atom, a lower alkyl group of 1 to 5 carbon atoms, or R ³ and R ⁴ represents a lower alkylene group which is bonded to the bond chain and may contain an oxygen atom or a nitrogen atom in its bonding chain).], And a physiologically acceptable salt thereof. Furthermore, the present invention is an immunomodulator containing, as an active ingredient, a benastatin derivative represented by the above formula (1) or a physiologically acceptable salt thereof.

The benastatin derivative represented by the above formula (1) and the physiologically acceptable salt thereof have a higher solubility in acidic, neutral and basic aqueous solutions than Benastatin C and D, and are extremely useful as pharmaceuticals. It is a compound. In the definition of the above formula (1), when R ³ and R ⁴ each represent a lower alkyl group having 1 to 5 carbon atoms, the lower alkyl group is, for example, a methyl group, an ethyl group, a propyl group, a butyl group or a pentyl group. And an alkyl group having 1 to 5 carbon atoms. The bonding chain optionally substituted lower alkylene group which may contain an oxygen atom or a nitrogen atom in, for example, _{- (CH 2) 3 -,} - (CH 2) 4 -, - (CH 2) 5
-,

[Chemical 4] _{- (CH 2) 2 -O- (} CH 2) 2 - is a lower alkylene group having 1 to 5 carbon atoms, and the like.

The substituent R ^{2 in the} above general formula [I] is --CO--
As the substituent R ² when representing (CH ₂ ) _n —CO ₂ H, for example, an oxalo group, a 2-carboxylacetyl group,
Examples thereof include a 3-carboxypropanoyl group, a 4-carboxybutyryl group and a 5-carboxypentanoyl group. The substituent R ² is —CO— (CH ₂ ) _n —NR ³ R ⁴
Examples of the substituent R ^{2 in the case} of are, for example, dimethylaminoacetyl group, diethylaminoacetyl group, diethylaminopropionyl group, butylaminoacetyl group, dimethylaminopropionyl group, dimethylaminobutyryl group,
Examples thereof include a pyrrolidinobutyryl group, a pyrrolidinoacetyl group, a piperazinoacetyl group and a morpholinoacetyl group. When the substituent R ² represents — (CH ₂ ) _{n + 1} —CO ₂ H, the substituent R ² is carboxymethyl group, 2-carboxyethyl group, 3-carboxypropyl group, 4-carboxybutyl group, 5 Examples include -carboxypentyl group and 6-carboxyhexyl group.

The substituent R ² is-(CH ₂ ) _{n + 1} --NR ³ R
^As the substituent R ² when ⁴ is shown, for example, an aminomethyl group, a dimethylaminomethyl group, a diethylaminomethyl group, a 2-aminoethyl group, a 2-methylaminoethyl group,
2-dimethylaminoethyl group, 2-ethylaminoethyl group, 2-diethylaminoethyl group, 2-pyrrolidinoethyl group, 2-piperidinoethyl group, 2-morpholinoethyl group, 2- (4-piperazino) ethyl group, 3- Aminopropyl group, 3-methylaminopropyl group, 3-dimethylaminopropyl group, 3-ethylaminopropyl group, 3-diethylaminopropyl group, 3-pyrrolidinopropyl group,
3-piperidinopropyl group, 3-morpholinopropyl group, 3- (4-piperazino) propyl group, 4-aminobutyl group, 4-methylaminobutyl group, 4-dimethylaminobutyl group, 4-ethylaminobutyl group , 4-diethylaminobutyl group, 4-pyrrolidinobutyl group, 4-piperidinobutyl group, 4-morpholinobutyl group, 4- (4-piperazino) butyl group, 5-aminopentyl group, 5-methylaminopentyl group, 5- Dimethylaminopentyl group,
5-ethylaminopentyl group, 5-diethylaminopentyl group, 5-pyrrolidinopentyl group, 5-piperidinopentyl group, 5-morpholinopentyl group, 5- (4-piperazino) pentyl group, 6-aminohexyl group, 6-methylaminohexyl group, 6-dimethylaminohexyl group,
Examples thereof include a 6-ethylaminohexyl group, a 6-diethylaminohexyl group, a 6-pyrrolidinohexyl group, a 6-piperidinohexyl group, a 6-morpholinohexyl group, and a 6- (4-piperazino) hexyl group.

The substituent R ² of the general formula [I] includes those mentioned above, and among the compounds of the general formula [I] having such a substituent R ² , a particularly preferred general formula [I]
The following compounds can be mentioned as the compounds of Compound number 1. 1- (3-dimethylaminopropanoyl) benastatin C 2. 1- (4-dimethylaminobutyryl) benastatin C 3. 1- (5-dimethylaminopentanoyl) benastatin C 4. 1- (3-carboxypropanoyl) benastatin C 5. 1- (4-carboxybutyryl) benastatin C 6. 1- (5-carboxypentanoyl) benastatin C 7. 1- (2-aminoethyl) benastatin C 8. 1- (2-methylaminoethyl) benastatin C 9. 1- (2-Dimethylaminoethyl) benastatin C 10.1- (3-aminopropyl) benastatin C 11.1- (3-methylaminopropyl) benastatin C 12.1- (3-dimethylaminopropyl) benastatin C 13 1- (3-Dimethylaminopropanoyl) Benastatin D 14.1- (4-Dimethylaminobutyryl) Benastatin D 15.1- (5-Dimethylaminopentanoyl) Benastatin D 16.1- (3-Carboxypropa Noyl) benastatin D 17.1- (4-carboxybutyryl) benastatin D 18.1- (5-carboxypentanoyl) benastatin D 19.1- (2-aminoethyl) benastatin D 20.1- (2-methyl Aminoethyl) benastatin D 21.1- (2-dimethylaminoethyl) Nasutachin D 22.1- (3- aminopropyl) Benasutachin D 23.1- (3- methylamino-propyl) Benasutachin D 24.1- (3- dimethylaminopropyl) Benasutachin D

The benastatin derivative of the general formula (1) may be in the form of a physiologically acceptable salt. Examples of such salts include inorganic acid salts such as hydrochloride, hydrobromide, sulfate and phosphate, or formate, acetate, fumarate, maleate, citrate and tartrate. , Organic acid salts such as lactate and methanesulfonate. Alternatively, sodium salt, potassium salt, magnesium salt,
Examples thereof include metal salts such as calcium salts and organic base salts such as quaternary ammonium salts and pyridinium salts.

The benastatin derivative represented by the general formula [I] is produced by the method described below. 1. In the general formula [I], the substituent R ² is —CO— (CH
₂ ) The Benastatin derivative which is _n- Y can be produced by any of the following methods. (1) R ² is —CO— (CH ₂ ) _n —CO ₂ H
To produce a benastatin derivative in which n is an integer from 2 to 5 :
Sodium hydroxide, potassium hydroxide, in a solvent such as hexane, benzene, toluene, ether, ethyl acetate, acetone, tetrahydrofuran, dioxane, pyridine, etc.
Sodium carbonate, potassium carbonate, sodium hydrogen carbonate,
Inorganic bases such as potassium hydrogen carbonate aqueous ammonia, 1,5-diazabicyclo [4,3,0] non-5-ene, 1,8-diazabicyclo [5,4,0] -7-undecene, pyridine, triethylamine, etc. Benastatin C or D using the organic base of

[Chemical 5] It can be obtained by reacting a dicarboxylic acid anhydride represented by the formula (n represents an integer of 2 to 5).

(2) R ² is --CO-(CH ₂ ) _n --C
Benastatin inducer, which is O ₂ H and n is an integer from 0 to 5
Production of conductor : a) Using a condensing agent such as N, N′-dicyclohexylcarbodiimide in the presence of a catalyst such as 4-dimethylaminopyridine in the same solvent as the above (1), if necessary, the following formula

Embedded image A compound represented by R ⁵ O ₂ C (CH ₂ ) _n CO ₂ H (wherein R ⁵ is a protecting group for an ester group, and n is an integer of 0 to 5) is converted to benastatin C or D. It can be obtained by reacting and removing the protecting group R ⁵ after the reaction. As the ester protecting group, a commonly used protecting group can be used, and elimination thereof can also be carried out by adopting a method known per se.

B) In a solvent similar to the above (1), if necessary, in the presence of a catalyst such as 4-dimethylaminopyridine,
Using the same base as the above (1), the following formula

Embedded image R ⁵ O ₂ C (CH ₂ ) _n COZ (wherein R ⁵ is a protecting group for an ester group, n is an integer of 0 to 5, Z is a halogen atom such as Cl, Br, I) It can also be obtained by reacting the compound represented by with Benastatin C or D, and deprotecting the protecting group R ⁵ after the reaction.

(3) R ² is --CO-(CH ₂ ) _n --N
R ³ production of R ⁴ a is Benasutachin derivative: a). Under the reaction conditions using the same solvent, catalyst and condensing agent as in the above (2) a), the following formula

[Chemical 8] R³R^FourN (CH₂)_nCO₂H (in the formula, R³, R^FourIs the same as above, and n is 0 to 5
The compound represented by the formula) is Benastatin C or
It can be obtained by reacting with D. R³R^Four
If a protecting group is required for N, it is introduced
Compound R ³R^FourN (CH₂)_nCO₂React with H, react
Then, it can be obtained by deprotecting the protecting group.
As the protecting group used here, as a protecting group for an amino group,
A commonly used protecting group can be used, and its elimination is also possible.
It can be carried out by a method known per se.

B). Under the reaction conditions using the same solvent, catalyst and base as in (b) of (2) above, the following formula

Embedded image R ³ R ⁴ N (CH ₂ ) _n COZ (wherein R ³ and R ⁴ are the same as defined above, n is an integer of 0 to 5 and Z is a halogen atom such as Cl, Br or I). Can also be obtained by reacting a compound represented by the formula) with Benastatin C or D. When a protecting group is required for R ³ R ⁴ N, a compound having a protecting group introduced R ³ R
It can be obtained by reacting ⁴ N (CH ₂ ) _n COZ, and deprotecting the protecting group after the reaction.

C). Under the same reaction conditions as a) or b) in the above (3), the following formulas are respectively used.

Embedded image W (CH ₂₎ in _n CO ₂ H (wherein, W is, Cl, Br, a halogen atom such as I, or a methanesulfonyloxy group,, p-toluenesulfonyloxy indicates an oxy group, etc., n represents 0 A compound represented by the formula:

Embedded image W (CH ₂ ) _n COZ (wherein W and n are the same as above, Z is Cl, B
It can also be obtained by reacting a compound represented by a halogen atom such as r or I) with benastatin C or D, and then reacting with an amino compound HNR ³ R ⁴ .

2. The benastatin derivative in which the substituent R ² in the general formula [I] is — (CH ₂ ) _{n + 1} —Y can be produced by any of the following methods. (1) In a solvent such as hexane, benzene, toluene, ether, ethyl acetate, acetone, tetrahydrofuran, dioxane, and pyridine, in the presence of diethylazodicarboxylate (DEAD) or triphenylphosphine,
The following formula

Reaction of a compound represented by Y- (CH ₂ ) _{n + 1} -OH (wherein Y is the same as defined above and n is an integer of 0 to 5) with benastatin C or D. Can be obtained by When a protecting group is required for Y in the above compound, the compound Y containing a protecting group
It can be obtained by reacting — (CH ₂ ) _{n + 1} —OH, and deprotecting the protecting group after the reaction.

(2) Hexane, benzene, toluene,
In aprotic solvents such as xylene, diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, diglyme, triglyme, etc., alkali metals such as sodium and potassium; methyllithium, ethyllithium, butyllithium, phenyllithium, methylmagnesium iodide, ethylmagnesium. bromide,
Organic metals such as phenyl magnesium bromide and phenyl magnesium iodide; organic hydrides such as lithium hydride, sodium hydride and potassium hydride; or lithium bis (trimethylsilyl) amide, lithium amide, sodium amide, potassium amide, lithium dicyclohexyl amide In the presence of a metal amide such as lithium diisopropylamide, the following formula

Embedded image Y- (CH ₂ ) _{n + 1} -V (Y is the same as above, n is an integer of 0 to 5, V is a halogen atom such as a fluorine atom, a base atom, a bromine atom, and a iodine atom. Or a compound represented by a methanesulfonyloxy group, a p-toluenesulfonyloxy group, or the like) is reacted with Benastatin C or D. When a protecting group is required for Y in the above compound, the compound Y- (CH ₂ ) _{n + 1} in which a protecting group is introduced is introduced.
It can be obtained by reacting —V and then deprotecting the protecting group.

(3) Under the same reaction conditions as in (1) or (2) of 2 above, the following formula

Embedded image V ₁ — (CH ₂ ) _{n + 1} —OH (In the formula, V ₁ represents a halogen atom such as Cl, Br, I, or a methanesulfonyloxy group, a p-toluenesulfonyloxy group, or the like; Represents an integer of 0 to 5), or a compound represented by the following formula:

Embedded image wherein V ₂ — (CH ₂ ) _{n + 1} —V ₃ (wherein V ₂ and V ₃ are halogen atoms such as Cl, Br and I, or a methanesulfonyloxy group, p-toluenesulfonyloxy group, etc., respectively). And n is an integer of 0 to 5) is reacted with Benastatin C or D, and then the amino compound HNR ³ R ⁴ is reacted.

The benastatin derivative of the above formula (1) can be obtained by the various production methods shown above. As the post-reaction treatment in each of the above-mentioned various production methods, the post-reaction treatment can be carried out according to a usual method depending on the various production methods. For example, if necessary, the precipitate is filtered off,
The reaction solution is dissolved in a solvent such as ethyl acetate, diethyl ether, tetrahydrofuran, dichloromethane, chloroform, toluene, benzene, xylene, and diluted hydrochloric acid, diluted sulfuric acid,
It is washed with an acid such as dilute nitric acid or a base such as dilute sodium hydroxide aqueous solution, dilute sodium carbonate aqueous solution, dilute potassium hydrogen carbonate aqueous solution, and dilute aqueous ammonia. After washing with an acid or a base, preferably, a post-reaction treatment can be carried out preferably by rapidly washing with an aqueous solution having the opposite property, that is, washing with a base or an acid, respectively, and further washing with water to remove the acid or the base.

As a purification method, an ordinary method can be adopted. For example, silica gel, alumina, diatomaceous earth, or the like, or a packing material obtained by modifying them with various coating agents is subjected to column chromatography of normal layer or reverse layer under normal pressure to several hundreds of atmospheric pressure, and hexane, toluene, ether. , Ethyl acetate, dichloromethane, chloroform, acetone, ethanol, methanol, water, acetic acid,
A method of eluting with dimethylsulfoxide, N, N-dimethylformamide, triethylamine, aqueous ammonia or the like or a mixed solvent thereof in an appropriate ratio, or recrystallization or reprecipitation using a similar solvent or a mixed solvent thereof in an appropriate ratio. Alternatively, these methods can be combined.

The benastatin derivative of the above formula (1) of the present invention can be converted into a physiologically acceptable salt, if necessary. The conversion into these salts can be carried out by converting into a salt such as the above-mentioned inorganic acid salt, organic acid salt, metal salt or organic base salt by a generally known method.

The benastatin derivative of the present invention has an immunomodulatory action, particularly a strong immunostimulatory action, and is therefore useful as an immunomodulator. As shown in the following test example, a benastatin derivative has an immunostimulatory action at a low concentration (eg, about 7 μg / ml or less) and an immunosuppressive action at a high concentration (eg, about 12 μg / ml or more) in a lymphocyte blast transformation reaction. Show. Moreover, this compound is not toxic. Therefore, the benastatin derivative is extremely useful as an immunomodulator. The benastatin derivative can be usually administered orally or parenterally such as intravenously, intracutaneously or intramuscularly. The dose varies depending on the administration subject, administration route, etc., but is usually 0.5 to 100 mg /
kg / day, preferably 1 to 50 mg / kg / day.

The formulation for administration includes conventional dosage forms. In the case of oral administration, tablets, granules, capsules and the like which are molded together with usual excipients such as starch are used. In the case of parenteral administration, a normal injection or the like formed by using physiological saline, a dissolving agent or the like is used.

[0024]

INDUSTRIAL APPLICABILITY As described above in detail, the present invention provides a novel physiologically active substance, benastatin derivative. The benastatin derivative of the present invention exhibits high solubility in water. For example, the sodium salt of Compound No. 5 is 0.23 mg / ml in water and shows a solubility 20 times or more that of Benastatin C. Furthermore, the benastatin derivative of the present invention exhibits an immunostimulatory action at a low concentration and an immunosuppressive action at a high concentration, and is therefore extremely useful as an immunomodulator.

The present invention will be described in more detail with reference to synthesis examples, formulation examples and test examples of the benastatin derivative of the present invention.

Synthesis Example 1 1- (4-Carboxybutyryl) Benastatin C (Compound
Synthesis of product number 5) Benastatin C (98 mg, 0.21 mmole) was dissolved in pyridine (7.0 ml), and glutaric anhydride (3
1 mg, 0.27 mmole) was added, and the mixture was stirred at room temperature for 3 days. Water was added to the reaction solution, extraction was performed twice with ethyl acetate, and the organic layer was washed with saturated brine. After drying over anhydrous sodium sulfate, the residue obtained by concentrating the organic layer under reduced pressure was purified by silica gel column chromatography (chloroform / tetrahydrofuran = 24/1 to 22/3) to give 1-
(4-Carboxybutyryl) Benastatin C (50m
g, yield 41%). Further, the fraction that is insufficiently purified is purified by preparative silica gel thin layer chromatography (developing solvent; chloroform: methanol = 10: 1),
1- (4-carboxybutyryl) benastatin C (11
mg, yield 9%) was obtained. ¹ H-NMR (200 MHz FT, TMS, CD ₃ O
D) 0.93 (3H, t, J = 6Hz) 1.33 to 1.47 (4H, m) 1.75 (6H, s) 1.62-1.86 (2H, m) 2.11 ( 2H, m) 2.52 (2H, t, J = 7Hz) 2.79 (2H, t, J = 8Hz) 3.01 (2H, t, J = 7Hz) 6.28 (1H, d, J = 2Hz) 6.71 (1H, d, J = 2Hz) 7.20 (1H, d, J = 2Hz) 7.60 (1H, d, J = 2Hz) 7.67 (1H, d, J = 9Hz) 8.23 (1H, d, J = 9Hz) 8.76 (1H, s) Mass spectrum FABmass 571.5 ([M + H] ⁺ ).

Example 2 1- (2-Dimethylaminoethyl) benastatin C
Synthesis of Compound No. 9) Benastatin C (72 mg, 0.16 mmole) was dissolved in tetrahydrofuran (4.0 ml), triphenylphosphine (47 mg, 0.18 mmole) was added, and azodicarboxylic acid ethyl ester (31 m) was added at room temperature.
A tetrahydrofuran solution (3.0 ml) of g, 0.18 mmole) was added dropwise. Then, a tetrahydrofuran solution (5.0 ml) of N, N-dimethylaminoethanol (17 mg, 0.19 mmole) was added dropwise, and the mixture was stirred at room temperature for 1 hour.
Stir for 4 hours. Water was added to the reaction solution, and ethyl acetate was added to 2
It was extracted twice and the organic layer was washed with saturated brine. After drying over anhydrous sodium sulfate, the residue obtained by concentrating the organic layer under reduced pressure is subjected to silica gel column chromatography (chloroform / acetic acid ethyl ester = 24/1 to 1/1, chloroform / methanol = 100/1 to 4/1). 1-
(2-Dimethylaminoethyl) Benastatin C (20m
g, yield 19%) was obtained. ¹ H-NMR (200 MHz FT, TMS, CDC
_{l 3: CD 3 OD = 10} : 1) 0.90 (3H, t, J = 7Hz) 1.21-1.47 (4H, m) 1.77 (6H, s) 1.53-1.92 (2H, m) 2.39 (6H, s) 2.73 (2H, t, J = 8Hz) 2.83 (2H, t, J = 5Hz) 4.17 (2H, t, J = 5Hz) 6 .41 (1H, d, J = 2Hz) 6.78 (1H, d, J = 2Hz) 6.95 (1H, d, J = 1Hz) 7.23 (1H, d, J = 1Hz) 7.62 (1H, d, J = 9Hz) 8.26 (1H, d, J = 9Hz) 9.61 (1H, s) Mass spectrum FABmass 528 ([M + H] ⁺ ).

[0027]

[Formulation Example] Example 3 Compound No. 5, 30 parts by weight, 120 parts of crystalline lactose, 147 parts of crystalline cellulose and 3 parts of magnesium stearate were mixed in a V-type mixer and compressed to give tablets of 300 mg each. .

[0028]

[Test Example] The test example below shows that the benastatin derivative has an immunomodulatory activity and exhibits no toxicity. Test Example 1 Immunization of benastatin derivative in lymphocyte blast transformation reaction
The test method of the regulatory effect of the benastatin derivative on the blastogenic reaction of regulatory active lymphocytes is as follows. 20% fetal bovine serum, 25 mM Hepes buffer, 100 μ for culture
RPMI 1640 medium supplemented with g / ml streptomycin and 100 units / ml penicillin G was used. Culture was performed on 96-well flat bottom microplates (COSTA
R). As mitogens, lipopolysaccharide (LPS) and concanavalin A (Con A) were used at final concentrations of 100 and 5 μg / ml, respectively.

Spleen cells are used in BALB / c mice (female,
> 20 weeks old) and remove the spleen to make a single cell suspension,
Red blood cells with hyper shock
Was removed and used for LPS as it was,
Nylon Fiber for T cell isolation for Con A
(Wako Pure Chemicals). 2 x 10 in each well ^Five
Add splenocytes and test compound at each dilution
The total volume was 0.2 ml, and this was cultured for 72 hours. End of culture
8 hours prior to completion, add 37 KBq / well [³H] -chi
Add midine and measure its uptake into cells
It was The effect is determined by the control of each test compound addition group.
Against [³[H] -Thymidine uptake ratio
(Japanese Society of Immunology, Immunological Experimental Procedures, 2267-2)
276). Mouse lymphocyte juveniles of benastatin derivatives
The effects on the rejuvenation reaction are shown in Tables 1 and 2. Benasta
Chin derivatives are lymphocyte blastogenic by LPS, Con A
The reaction was very strongly accelerated at low concentrations.

[0030]

Action of TABLE 1 LPS and Compound No. definitive mice lymphocyte transformation reaction by Con A 5 (Na salt) ─────────────────────── ────────── Compound concentration index (%) number (μg / ml) LPS Con A ──────────────────────── ───────── 5 (Na salt) 0.1 155.8 159.8 0.33 396.3 293.8 1 982.6 947.2 3.3 863.0 1320.1 10 22.6 13.3 33 2.0 1.7 100 1.8 1.6 Control 100.0 100.0 ──────────── ─────────────────────

[0031]

Action of Table 2 LPS and Compound No. definitive mice lymphocyte transformation reaction by Con A 9 (HCl salt) ─────────────────────── ────────── Compound concentration index (%) number (μg / ml) LPS Con A ──────────────────────── ───────── 9 (HCl salt) 0.1 111.1 141.4 0.33 189.4 112.0 1 291.3 239.9 3.3 265.3 268.2 10 72.9 87.5 33 4.2 8.3 100 2.3 0.8 Control 100.0 100.0 ──────────── ─────────────────────

 ─────────────────────────────────────────────────── ─── Continued Front Page (72) Inventor Takaaki Aoyagi 3-3-6 Motokugenuma, Fujisawa City, Kanagawa Prefecture (72) Inventor Tomio Takeuchi 5-1-11-701A, Higashigotanda, Shinagawa-ku, Tokyo

Claims (2)

[Claims] 1. The following formula (1): Wherein, R ¹ is -CH = CH- or -CH ₂ CH ₂ - indicates, R ² is -CO- (CH _{2) n} -Y or - (C
Shows the H _{2) n +} 1 -Y. Where n is an an integer of 0 to 5, Y is -CO ₂ H or NR ³ R ⁴ (R ^3, R ⁴ are each a hydrogen atom, a lower alkyl group of 1 to 5 carbon atoms, or R ³ and R ⁴ represents a lower alkylene group which is bonded to the bond chain and may contain an oxygen atom or a nitrogen atom in its bonding chain.]] And a physiologically acceptable salt thereof. 2. An immunomodulator comprising the benastatin derivative according to claim 1 or a physiologically acceptable salt thereof as an active ingredient.