JPH075631B2 - Glutathione derivative - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION “Industrial field of application” The compound of the present invention has an effect of suppressing liver damage and is useful as a therapeutic agent for liver diseases.

"Problems to be solved by conventional techniques and inventions" Glutathione is a thiol compound that is present in many organisms, and is present in high concentration in the liver, which is particularly important for the detoxification function. It is well known that it plays an important role in detoxification metabolism and regulation of redox state. Although it is widely used in clinical medicine for the treatment of various liver diseases, poisoning due to drugs and heavy metals, allergic diseases, etc., a large amount of glutathione is currently used. In the treatment of various liver diseases, the purpose is to maintain the level of glutathione in hepatocytes and improve detoxification and metabolic functions.However, glutathione is not taken up by the liver, and is completely hydrolyzed by other organs and then transported to the membrane as an amino acid. Then, it enters the liver cytoplasm from the blood and glutathione is resynthesized. This is extremely disadvantageous for the purpose of clinically administering glutathione and directly increasing / maintaining glutathione level in hepatocytes.

"Means and Actions for Solving Problems" In order to solve these problems, a compound represented by the general formula [I] in which the N-terminal amino group of glutathione and its ester compound is phosphoramidated is synthesized, As a result of examining their uptake into the liver, it was found that the compound of the present invention has a liver damage suppressing effect superior to that of glutathione.

(In the formula, R ¹ and R ² are the same or different and are a lower alkyl group, a hydroxy group, a lower alkoxy group, a phenyl group,
A phenyloxy group, a phenyl lower alkyl group or a phenyl lower alkoxy group is shown, and a phenyl ring of a phenyl group, a phenyloxy group, a phenyl lower alkyl group or a phenyl lower alkoxy group is a lower alkyl group, a hydroxy group, a lower alkoxy group, a nitro group or a halogen group. It may be substituted with one or more groups selected from atoms.

R ³ and R ⁴ are the same or different and each represents a hydrogen atom, a lower alkyl group, a phenyl group or a phenyl lower alkyl group, and the phenyl ring of the phenyl group and the phenyl lower alkyl group is further a lower alkyl group, a hydroxy group or a lower alkoxy group. It may be substituted with one or more groups selected from groups, nitro groups and halogen atoms.

Y represents oxygen or sulfur. same as below. Each group will be described in more detail. The lower alkyl group means an alkyl group having 1 to 6 carbon atoms such as a methyl group, an ethyl group, a propyl group and a hexyl group, and the lower alkoxy group means a methoxy group. Group, ethoxy group,
An alkoxy group having 1 to 6 carbon atoms such as propoxy group and hexyloxy group is shown, and a lower alkanoyloxy group means 2 to 6 carbon atoms such as acetyloxy group, propionyloxy group and hexanoyloxy group. Indicates an alkanoyloxy group having, a halogen atom is fluorine,
Indicates chlorine, bromine and iodine.

The method for producing the compound of the present invention will be briefly described in the following figure and is shown by the following method.

(1) A method of using oxidized glutathione or an ester thereof as a starting material.

(2) A method using SH group-protected glutathione or its ester as a starting material.

In the above reaction formula, X represents a halogen atom, and R ⁵ represents a protecting group such as a trityl group. The protective group such as trityl group can be removed by a conventional method.

The compound represented by the formula [I] can be converted to a pharmaceutically acceptable salt such as sodium salt and potassium salt, if necessary.

"Example" Example 1. Production of N- (dibenzylphosphoryl) glutathione dibenzyl ester (i) Production of N, N'-bis (dibenzylphosphoryl) oxidized glutathione tetrabenzyl ester Dibenzyl hydrogen phosphite (9.23) A solution of sulfuryl chloride (2.6 ml) in anhydrous carbon tetrachloride (10 ml) was added dropwise to the solution of g) in anhydrous carbon tetrachloride (30 ml) under nitrogen gas at 0 ° C or lower with stirring, and then the mixture was stirred at room temperature for 2 hours. The above solution and triethylamine (2.5m
The solution of l) in DMF (10 ml) was simultaneously added dropwise to a solution of oxidized glutathione tetrabenzyl ester (7.79 g) in DMF (30 ml) at 0 ° C. or lower, and then the mixture was stirred under ice cooling for 2 hours and further at room temperature for 16 hours. Chloroform was added to the reaction mixture, N hydrochloric acid, water,
It is washed with a saturated aqueous solution of sodium hydrogencarbonate, water and saturated saline in this order, dried over anhydrous magnesium sulfate and concentrated under reduced pressure.
The residue was purified by silica gel column chromatography to give the title compound.
Obtain 8.93 g (78%).

Melting point 164-165.5 ° C (ethyl acetate) ▲ [α] ²⁵ _D ▼ + 6.93 ° (c = 2.0, CHCl ₃ ) IR (KBr, cm ^-1 , the same unless otherwise specified): 1728,1634,1213,998 (Ii) Production of N- (dibenzylphosphoryl) glutathione dibenzyl ester N, N'-bis (dibenzylphosphoryl) oxidized glutathione tetrabenzyl ester (2.
A mixture of 24 g), dioxane (40 ml), water (1 ml) and tri-n-butylphosphine (1.1 ml) is stirred at room temperature for 14 hours. After concentration under reduced pressure, the residue was purified by silica gel column chromatography to obtain 1.86 g (83%) of the title compound.

Melting point 105-107 ° C (ethyl acetate-n-hexane) ▲ [α] ²⁵ _D ▼ -19.1 ° (c = 2.0, CHCl ₃ ) IR: 1727,1633,1212,1003. Can be obtained.

-N- (diethylphosphoryl) glutathione dibenzyl ester, melting point 127-129 ° C (ethyl acetate-n-hexane) ▲ [α] ²⁵ _D ▼ -21.2 ° (c = 2.0, CHCl ₃ ) IR: 1726,1636,1222, 1029. ・ N- (dipropylthiophosphinyl) glutathione
Diethyl ester IR: 1732,1629. N- (dibutylphosphoryl) glutathione t-butyl 4-nitrobenzyl ester (R ³ = t-butyl, R ⁴
= 4-nitrobenzyl) IR: 1727,1634,1205,1009. ・ N- (diphenylphosphinyl) glutathione benzyl ethyl ester (R ³ = benzyl, R ⁴ = ethyl) IR: 1728,1639. ・ N- (Diphenylphosphoryl) glutathione dibenzyl ester IR: 1730,1633,1230,1005. ・ N- [bis (4-iodobenzyl) phosphoryl] glutathione di-t-butyl ester IR: 1728,1629,1210,1005. N- (O-benzyl-P-phenethylphosphonyl) glutathione ethyl cyclohexyl ester (R ¹ = benzyloxy, R ² = phenethyl, R ³ = ethyl, R ⁴ = cyclohexyl) IR: 1732,1636,1212,1008 . eXAMPLE 2. N-(dibenzyl phosphoryl) glutathione-benzyl ethyl ester (R ³ = benzyl, R ⁴ = ethyl) preparation of (i) N-(dibenzyl phosphoryl) -S- (trityl) glutathione Emissions benzyl ethyl ester (R ³ = benzyl, R ⁴ = ethyl) prepared dibenzyl hydrogen phosphite phosphite of (2.88 g), sulfuryl chloride (0.8 ml), carbon tetrachloride (30ml) of Example 1 than the same manner as (i) A carbon tetrachloride solution of dibenzyl phosphoric acid chloride is prepared by the procedure of. This solution and triethylamine (1.4 ml) were added to S- (trityl) glutathione benzyl ethyl ester (3.0 g) in DMF (10 m
l) The solution was added dropwise to the solution at 0 ° C. or lower at the same time, and the mixture was further stirred overnight under ice cooling at room temperature for 2 hours. The insoluble matter is removed, and the solution is concentrated under reduced pressure. The residue is dissolved in ethyl acetate, washed with N hydrochloric acid, 5% aqueous sodium hydrogen carbonate solution and water in this order, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography to give 3.80 g (91%) of the title compound as an oil.

・ ▲ [α] ²⁵ _D ▼ −4.3 ° (c = 1.0, acetone) IR (film, cm ⁻¹ ): 1739,1656,1209,1003. (Ii) N- (dibenzylphosphoryl) glutathione benzyl ethyl ester Production N- (dibenzylphosphoryl) -S-obtained in (i)
To a solution of (trityl) glutathione benzyl ethyl ester (3.71 g) in DMF (20 ml) is added a solution of silver nitrate (2.04 g) and pyridine (0.97 ml) in methanol (100 ml), and the mixture is stirred at room temperature for 3.5 hours in the dark. Remove the deposit,
This is suspended in a mixed solution of DMF (20 ml) and methanol (20 ml), 2-mercaptoethanol (4.2 ml) is added, and the mixture is stirred at room temperature for 1 hr. The precipitate is removed and the liquid is concentrated under reduced pressure. The residue is dissolved in ethyl acetate, washed with water, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue is recrystallized to give 2.25 g (82%) of the title compound.

Melting point 131-132 ° C (ethyl acetate-n-hexane) ▲ [α] ²⁵ _D ▼ -10.3 ° (c = 1.0, DMSO) IR: 1729,1631,1235,1020. In the same manner as in Example 2 below. The compound can be obtained.

· N-(diethylphosphoryl) glutathione-benzyl 4-methoxybenzyl ester (R ³ = benzyl, R ⁴ =
4-methoxybenzyl) IR: 1725,1659,1222,1015. N- (O-ethyl-P-phenylthiophosphoryl) glutathione diethyl ester (R ¹ = ethoxy, R ² = phenyl) IR: 1735,1655, 1225,1033. N- [bis (4-bromobenzyl) phosphoryl] glutathione diethyl ester IR: 1735,1631,1227,1011. Example 3. Preparation of N- (dibenzylphosphoryl) glutathione (i) N- ( Preparation of dibenzylphosphoryl) -S- (trityl) glutathione From dibenzylhydrogenphosphite (2.30 g), sulfuryl chloride (0.65 ml) and carbon tetrachloride (20 ml), the same procedure as in (i) of Example 1 was performed. Prepare a solution of dibenzyl phosphate chloride in carbon tetrachloride. This solution and triethylamine (2.2 ml) were simultaneously added dropwise to a solution of S- (trityl) glutathione (2.20 g) in DMF (20 ml) at 0 ° C or lower, and the mixture was stirred under ice cooling for 3 hours. The insoluble matter is removed, and the solution is concentrated under reduced pressure. Chloroform and water are added to the residue, and N hydrochloric acid is added to acidify the residue, followed by extraction with chloroform. The extract is washed with water and saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residual oil is crystallized in ether to give 2.01 g (62%) of the title compound.

Melting point 170-174 ° C (decomposition) ▲ [α] ²⁵ _D ▼ -2.9 ° (c = 1.0, CHCl ₃ ) IR: 3276,1647,1517,1209. (Ii) Production of N- (dibenzylphosphoryl) glutathione ( N- (dibenzylphosphoryl) -S- obtained in i)
A solution of (trityl) glutathione (1.62 g) in DMF (10 ml) is added with a solution of silver nitrate (1.02 g) and pyridine (0.5 ml) in methanol (45 ml), and the mixture is stirred at room temperature for 1 hour while protected from light. The precipitate is taken, suspended in a mixed solution of DMF (15 ml) and methanol (15 ml), 2-mercaptoethanol (2.1 ml) is added, and the mixture is stirred at room temperature for 1 hr. The precipitate is removed and the liquid is concentrated under reduced pressure. The residue is solidified in acetone to give 0.59 g (52%) of the title compound.

Melting point 82-85 ° C. (decomposition) ▲ [α] ²⁵ _D ▼ -14.3 ° (c = 0.5, DMSO) IR: 3256,1636,1527,1214. The following compound can be obtained in the same manner as in Example 3. it can.

・ N- (diethylphosphoryl) glutathione Melting point 89-92 ° C (decomposition) ▲ [α] ²⁵ _D ▼ -10.8 ° (c = 1.0, DMSO) IR: 3244,1635,1519,1213. ・ N- (diphenylthio) Phosphinyl) glutathione IR: 3267,1632,1510.-N- [bis (4-nitrobenzyl) phosphoryl] glutathione IR: 3295,1638,1520,1232.-N- (phosphoryl) glutathione IR: 3250,1640, 1515,1210. Pharmacological test To investigate the inhibitory effect on liver damage, a drug that causes liver damage is administered to animals, and the effect of suppressing experimental liver damage caused thereby is examined. Drugs that cause experimental liver damage include carbon tetrachloride, thioacetamide, bromobenzene, paracetamol, D-galactosamine, etc. Particularly, carbon tetrachloride liver damage is toxic because the carbon-chlorine bond is cleaved by cytochrome P-450. Strong free radical (C
Cl ₃ ) and these free radicals are thought to bind to thiol groups of hepatocyte membrane proteins and promote lipid peroxidation of the membrane to cause damage [Biochem. Pharmacol.
21 , 49 (1972), 25 , 2163 (1976)].

In the present invention, in order to search the liver damage inhibitory effect, carbon tetrachloride is used as a drug causing experimental liver damage, and the liver damage inhibitory effect of the present compound is determined by serum transaminase (S-GO).
T and S-GPT) were used as indicators.

An experiment was conducted using N- (dibenzylphosphoryl) glutathione benzyl ethyl ester (R ³ = benzyl, R ⁴ = ethyl) as a representative example of the compound of the present invention and glutathione as a comparative drug.

Compound I: Glutathione (comparative drug) Compound II: N- (dibenzylphosphoryl) glutathione
Benzyl ethyl ester (Compound of the present application, R ³ = benzyl, R ⁴ = ethyl) Experimental example Male Wistar rat weighing 170 to 200 g was used as a group 3-5
The animals were used as animals after being fasted for 17 hours. Compounds I and II were orally administered at 300 mg / kg body weight. 60 minutes after the administration of each compound, carbon tetrachloride was intraperitoneally administered at 0.25 ml per 1 kg body weight (5 ml per 1 kg body weight as an olive oil solution). As a control, 5 ml of olive oil solution per 1 kg of body weight was intraperitoneally administered. Then, 24 hours after the administration of carbon tetrachloride, serum transaminase was measured.

As a result, as shown in the table, in comparison with the control group, a slight tendency of inhibition was observed in the glutathione (Compound I) administration group, whereas in the compound II, the significance of S-GOT and S-GPT was significant. Suppression was recognized.

"Effects of the Invention" The compound of the present invention, which is a glutathione derivative, has a liver damage suppressing effect superior to that of glutathione, and is useful as a therapeutic agent for liver damage.

Claims (1)

[Claims] 1. A compound represented by the general formula [I] and salts thereof. (In the formula, R ¹ and R ² are the same or different and are a lower alkyl group, a hydroxy group, a lower alkoxy group, a phenyl group,
A phenyloxy group, a phenyl lower alkyl group or a phenyl lower alkoxy group is shown, and a phenyl ring of a phenyl group, a phenyloxy group, a phenyl lower alkyl group or a phenyl lower alkoxy group is a lower alkyl group, a hydroxy group, a lower alkoxy group, a nitro group or a halogen group. It may be substituted with one or more groups selected from atoms. R ³ and R ⁴ are the same or different and each represents a hydrogen atom, a lower alkyl group, a phenyl group or a phenyl lower alkyl group, and the phenyl ring of the phenyl group and the phenyl lower alkyl group is further a lower alkyl group, a hydroxy group or a lower alkoxy group. It may be substituted with one or more groups selected from groups, nitro groups and halogen atoms. Y represents oxygen or sulfur. )