JPS62909B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to an epoxysuccinic acid diester compound, and more particularly to an epoxysuccinic acid diester compound whose thiol group inhibits the activity of a protease involved in the expression of the activity. As a result of various studies, the present inventors discovered that the thiol group of a trans-epoxysuccinic acid diester compound strongly inhibits the activity of proteases involved in the expression of its activity, and that it has extremely low toxicity. Completed the invention. The present invention will be explained in detail below. The object compound of the present invention has the general formula (In the formula, R is a benzyl group having 1 to 3 substituents selected from "halogen atom, methyl group, methoxy group and trifluoromethyl group", piperonyl group, 1-naphthylmethyl group, "halogen atom and 1-naphthylmethyl group, 3-phenyl group having 1 to 2 substituents selected from ``methyl group''
It represents a 2-propenyl group, a furfuryl group, a 2-tenyl group, or a tetrahydrofurfuryl group. ) is an epoxysuccinic acid diester compound represented by Here, the halogen atoms are fluorine, chlorine, bromine, and iodine. Also, benzyl group or 1-
When there are two or more substituents of the naphthylmethyl group,
They may be the same or different. A compound represented by the general formula () [hereinafter abbreviated as compound ()]. ] can be produced by the following method. That is, epoxysuccinic acid and an alcohol represented by the general formula R-OH () (wherein R has the same meaning as above) are reacted by a conventional method in the presence of an acidic catalyst such as sulfuric acid, or epoxysuccinic acid Epoxysuccinic acid dihalide obtained by treating with a halogenating agent such as phosphorus pentachloride, oxalyl chloride or thionyl chloride and the alcohol ()
to esterify epoxysuccinic acid.
After distilling off the solvent of the obtained reaction solution, the solidified substance is n-hexane, n-hexane-acetone, or n-hexane.
Recrystallize using a suitable solvent system such as -hexane-ethyl ether, and prepare the oil by fractional distillation under reduced pressure or by using silica gel as a carrier and methanol,
Compound () can be obtained by separation and purification by column chromatography using ethyl acetate, chloroform, benzene, etc., or an appropriate mixture thereof as a developing solvent. In addition, all compounds () in this specification are trans isomers. The compound (), which is the object of the present invention, strongly inhibits the activity of proteolytic enzymes such as papain, huicin, bromelain, bromelain, and cathepsin B, whose thiol group is thought to be involved in the expression of their activity, and furthermore, Extremely low toxicity. In order to demonstrate that compound () strongly inhibits the activity of these enzymes, a test example using papain is shown below. Test example 0.25 ml of a cysteine solution (PH6.8) adjusted to a 40 mmol concentration with a 20 mmol disodium ethylenediaminetetraacetic acid solution and 0.25 ml of a 10% dimethyl sulfoxide solution of compound () were mixed with an aqueous papain solution (80 μg/ ml, manufactured by Sigma, 2
Add to 0.5 ml of 1% milk casein solution [33 mmol concentration phosphate buffer (PH
6.8)] was added, and the mixture was further heated at 40°C for 10 minutes to react, and then 5 ml of a 440 mmol trichloroacetic acid solution was added to stop the reaction. After that, the absorbance A of the solution at 280 nm was measured, and at the same time, as a control, 10
Absorbance B was measured using a % dimethyl sulfoxide solution, and the activity inhibition rate was calculated from B-A/B×100. The amount of the compound showing 50% inhibition of activity by this method was defined as ID ₅₀ and shown in Table 1.

【table】

[Table] Compound () does not inhibit the caseinolytic activity of serine proteases typified by trypsin and acidic proteases typified by pepsin. That is, compound () specifically inhibits only the activity of the protease in which the thiol group is involved, and does not inhibit any other activities. Next, the present invention will be explained with reference to Examples. Example 1 A solution of 1.68 g of epoxysuccinic acid dichloride dissolved in 10 ml of benzene was added dropwise to a solution of 3.74 g of p-bromobenzyl alcohol and 1.58 g of pyridine dissolved in 50 ml of benzene under ice-cooling and stirring, and after stirring for 30 minutes, a solution was formed. Pyridine hydrochloride was removed, the benzene layer was washed with water, and after drying over anhydrous sodium sulfate, the benzene was distilled off, and the residue was recrystallized from n-hexane-acetone to produce colorless needle-shaped di-p-bromobenzyl epoxysuccinate. 3.43 g of ester was obtained. mp 124-125℃ Yield 73% Example 2 In the same manner as in Example 1, 2.84 g of p-chlorobenzyl alcohol and epoxysuccinic acid dichloride were added.
Colorless acicular epoxysuccinic acid di-p- from 1.68g
2.9 g of chlorobenzyl ester was obtained. mp 117°C Yield 76% Example 3 In the same manner as in Example 1, 4.68 g of p-iodobenzyl alcohol and epoxysuccinic dichloride were added.
Colorless scaly epoxysuccinic acid di-p from 1.68g
-3.6g of iodobenzyl ester was obtained. Yield 64% mp 137-139°C Example 4 In the same manner as in Example 1, 4.68 g of o-iodobenzyl alcohol and epoxysuccinic acid dichloride were added.
Colorless acicular epoxysuccinic acid di-o- from 1.68g
3.8 g of iodobenzyl ester was obtained. Yield 67% mp 111-112°C Example 5 In the same manner as in Example 1, 2.52 g of m-fluorobenzyl alcohol and epoxysuccinic acid dichloride were added.
Colorless acicular epoxysuccinic acid di-m- from 1.68g
2.5 g of fluorobenzyl ester was obtained. Yield 72% mp 37-38°C Example 6 In the same manner as in Example 1, 2.44 g of p-methylbenzyl alcohol and epoxysuccinic dichloride were added.
Colorless acicular epoxysuccinic acid di-p- from 1.68g
2.8 g of methylbenzyl ester was obtained. Yield 82% mp 77°C Example 7 In the same manner as in Example 1, 2.76 g of p-methoxybenzyl alcohol and epoxysuccinic acid dichloride were added.
Colorless scaly epoxysuccinic acid di-p from 1.68g
-2.9 g of methoxybenzyl ester was obtained. Yield 78% mp 82-84℃ Example 8 Piperonyl alcohol was prepared in the same manner as in Example 1.
From 3.04 g and 1.68 g of epoxysuccinic acid dichloride, 3.1 g of epoxysuccinic acid dipiperonyl ester in the form of a colorless powder was obtained. Yield 78% mp 129-132°C Example 9 In the same manner as in Example 1, colorless acicular epoxysuccinic acid di-
3.2 g of 2,4-dichlorobenzyl ester was obtained. Yield 71% mp 60℃ Example 10 In the same manner as in Example 1, cinnamyl alcohol
2.6 g of colorless acicular epoxy succinic acid disinnamyl ester was obtained from 2.68 g of epoxy succinic acid dichloride. Yield 71% mp 165°C Example 11 In the same manner as in Example 1, 2.9 g of colorless needle-shaped epoxysuccinic acid di-1-naphthyl methyl ester was obtained from 3.16 g of 1-naphthylmethanol and 1.68 g of epoxysuccinic acid dichloride. Ta. Yield 70% mp 105-107°C In the same manner as in Example 1, the following epoxysuccinic acid diester was obtained using the corresponding alcohol and epoxysuccinic acid dichloride. Epoxysuccinic acid di-m-chlorobenzyl ester Epoxysuccinic acid di-o-chlorobenzyl ester Epoxysuccinic acid di-3-chloro-4-brombenzyl ester Epoxysuccinic acid di-2,5-dichlorobenzyl ester Epoxysuccinic acid di-2,5-dichlorobenzyl ester Di-2-bromo-4-iodobenzyl ester Epoxysuccinic acid di-2-bromo-3-methylbenzyl ester Epoxysuccinic acid di-1-(5'-bromo)naphthylmethyl ester Epoxysuccinic acid di-1-(2) '-Chlor) naphthyl methyl ester Epoxysuccinic acid di-1-(4',5'-dimethyl) naphthyl methyl ester Example 12 In a solution of 2.04 g of tetrahydrofurfuryl alcohol and 1.58 g of pyridine dissolved in 50 ml of benzene,
1.68g of epoxysuccinic acid dichloride and 10% of benzene
ml of the solution was added dropwise under ice-cooling and stirring for 30 minutes. After stirring for 30 minutes, the resulting pyridine hydrochloride was removed, the benzene layer was washed with water, dried over anhydrous sodium sulfate, and the benzene was distilled off. The residue was fractionally distilled under reduced pressure to obtain colorless oily epoxysuccinic acid ditetrahydrofurfuryl ester.
2.4g was obtained. Yield 80% bp 221-224℃ (6mmHg) Example 13 Furfuryl alcohol 1.96g and pyridine 1.58g
A solution of 1.68 g of epoxysuccinic acid dichloride dissolved in 10 ml of benzene was added dropwise to a solution of 1.68 g of epoxysuccinic acid dichloride dissolved in 50 ml of benzene under ice-cooling and stirring. After stirring for 30 minutes, the formed pyridine hydrochloride was removed and the benzene layer was washed with water.
After drying with anhydrous sodium sulfate, benzene is distilled off. The residue was purified by silica column chromatography using n-hexane-acetone (10:1) as a developing solvent to obtain 2.1 g of epoxysuccinic acid difurfuryl ester as a colorless oil. Yield 72% MS m/e: 292 (M ⁺ ) IR ν ^neat _nax cm ^-1 : 1750, 1185 (ester) 91
8
(Epoxy) NMR (CDCl ₃ ) δ: 3.65 (2H, s,

[Formula]), 5.60 (4H, s, 2×-o-CH ₂ -), 6.25-6.45 (4H, m,

[Formula]), 7.30-7.43 (2H, m,

[Formula]) Example 14 In the same manner as in Example 13, 2.28 g of 2-thiophene methanol and 1.68 g of epoxysuccinic acid dichloride were added.
2.5 g of epoxysuccinic acid dithenyl ester was obtained as a pale yellow oil. Yield 77% MS m/e: 324 (M ⁺ ) IR ν ^neat _nax cm ^-1 : 1750, 1185 (ester),
890
(Epoxy) NMR (CDCl ₃ ) δ: 3.75 (2H, s,

[Formula]), 5.41 (4H, s, 2×-O-CH ₂ -), 6.95-7.85 (6H, m,

[Formula]) Example 15 Colorless oily epoxysuccinic acid di-m-trifluoromethylbenzyl ester was prepared from 3.52 g of m-trifluoromethylbenzyl alcohol and 1.68 g of epoxysuccinic acid dichloride in the same manner as in Example 13.
3.3g was obtained. Yield 74% IR ν ^neat _nax cm ^-1 : 1755, 1185 (ester) 89
Five
(Epoxy) NMR (CDCl ₃ ) δ: 3.68 (2H, s,

[Formula]), 5.17 (4H, s, 2×-O-CH ₂ -), 7.43 (8H, br s,

[Formula]) Example 16 In the same manner as in Example 13, a colorless oily epoxysuccinic acid di-
2.9 g of 3,5-dimethylbenzyl ester was obtained. Yield 79% MS m/e: 368 (M ⁺ ) IR ν ^neat _nax cm ^-1 : 1750, 1185 (ester),
895
(Epoxy) MMR: δ ^CDCl3 2.28 (12H, s, 4×-CH ₃ ),
3.66(2H,s,

[Formula]), 5.07 (4H, s, 2×-O-CH ₂ -), 6.88 (6H, s,

[Formula]) In the same manner as in Example 13, the following epoxysuccinic acid diester was obtained using the corresponding alcohol and epoxysuccinic acid dichloride as raw materials. Epoxysuccinic acid di-O-fluorobenzyl ester Epoxysuccinic acid di-O-methylbenzyl ester Epoxysuccinic acid di-2,4,6-trimethylbenzyl ester

Claims (1)

[Claims] 1. General formula (In the formula, R is a benzyl group having 1 to 3 substituents selected from "halogen atom, methyl group, methoxy group and trifluoromethyl group", piperonyl group, 1-naphthylmethyl group, "halogen atom and methyl 1-naphthylmethyl group, 3-phenyl-2 having 1 to 2 substituents selected from
- Indicates a propenyl group, furfuryl group, 2-thenyl group or tetrahydrofurfuryl group. ) An epoxysuccinic acid diester compound represented by