WO2001064661A1

WO2001064661A1 - Antioxidants

Info

Publication number: WO2001064661A1
Application number: PCT/JP2001/001347
Authority: WO
Inventors: Kazuo Nishimura; Kazumasa Takao; Hiroshi Suhara; Hideo Utsumi
Original assignee: Santen Pharmaceutical Co., Ltd.
Priority date: 2000-02-29
Filing date: 2001-02-23
Publication date: 2001-09-07
Also published as: AU2001234148A1

Abstract

The invention provides excellent antioxidants. Compounds of the general formula (1) or pharmacologically acceptable salts thereof are excellent in antioxidant activity and therefore useful as remedies for cataract and ischemic heart diseases due to oxidative stress, wherein R1, R2 and R3 are each independently hydrogen or lower alkyl; R4 is a carboxylic acid group or an ester or amide thereof; m is 0 or 1; and n is 1 or 2.

Description

Description Antioxidant Technical field

The present invention relates to an antioxidant containing a cyclic disulfide derivative as an active ingredient, and is useful as a therapeutic agent for cataract, ischemic disease, and the like caused by oxidative stress. Background art

Diseases involving oxidative stress include ischemic diseases such as myocardial infarction and cerebral infarction, cataracts, lung diseases such as emphysema and asthma, and nervous systems such as Alzheimer's syndrome and Parkinson's disease. Various diseases such as diseases, diabetes, and viral diseases are included.

By the way, daltathione (GSH) is widely present in living organisms and is an important protective factor against oxidative damage processes in living organisms. And, in many diseases in which the involvement of oxidative stress has been discussed, a decrease in GSH in that tissue has been observed. Lipoic acid, a vitamin-like substance having an S—S bond in the molecule, is an antioxidant that improves the oxidative damage process in vivo by increasing GSH in cells. Has been reported (I. Maitra et al., Free Radical Biology & Medicine, 18_ (4), 823-829 (1995)), but due to its strong irritation, It is not suitable for administration, and the effect of increasing GSH is not always sufficient.

On the other hand, the-part of the cyclic disulfide derivative which is the active ingredient of the present invention is a known substance, and Japanese Patent Application Laid-Open No. 2-132871 discloses the treatment of liver disease or autoimmunity such as rheumatoid arthritis. Cure of disease Effectiveness as a therapeutic agent is disclosed in Japanese Patent Application Laid-Open No. 4-208271, which has an effect of promoting the production of interferon, and Japanese Patent Application Laid-Open No. 11-255,648. With respect to the above-mentioned cyclic disulfide derivatives in which the angiogenesis inhibitory action is described, it is a very interesting subject to study further new pharmacological actions and to create new derivatives.

Disclosure of the invention

The present inventors conducted various pharmacological tests and found that the cyclic disulfide derivative of the present invention has an effect of remarkably increasing intracellular daltathione (GSH) and is useful as an antioxidant. The present invention has been found. The present invention is an antioxidant comprising a compound represented by the general formula [1] or a pharmaceutically acceptable salt thereof as an active ingredient.

Wherein R, R 2 and R ₃ are the same or different and each represents a hydrogen atom or a lower alkyl group, R ₄ represents a carboxylic acid or its ester or amide, m represents 0 or 1, and n represents Indicates 1 or 2, respectively. In the compound represented by the general formula [1], an ester is a lower alkyl such as methyl ester, ethyl ester, propyl ester, butyl ester, hexyl ester, isopropyl ester, isobutyl ester, and tert.-butyl ester. Ester; hydroxy lower alkyl ester such as hydroxymethyl ester, hydroxy shester ester, hydroxy propyl ester (this hydroxy group may be protected by a protecting group described later); cyclopropyl ester, cyclohexyl ester Cyclic alkyl esters having 3 to 6 carbon atoms, such as xyl esters; lower alkanes such as acetylaminomethyl ester, acetylaminoethyl ester, propionylaminomethyl ester, and propionylaminoethyl ester; Those generally used as esters of carboxylic acids such as lower amino lower alkyl esters; phenyl lower alkyl esters such as benzyl esters; phenyl esters; methoxy phenyl esters; and dandanyl esters.

In the compound represented by the general formula [1], amide is amide with ammonium; amide with lower alkylamine such as methylamine, dimethylamine, ethylamine and the like. De; Shows those commonly used as amides of ruboric acid, such as amides with phenyl lower alkylamines such as benzylamine.

—A good example of general formula [1] is R! R 2 and R ₃ are the same or different and each represents a hydrogen atom or a lower alkyl group such as a methyl group, an ethyl group or a butyl group; and R ₄ represents a lower alkyl ester such as a methyl ester or an ethyl ester, a hydroxymethyl ester, Amides with hydroxy lower alkyl esters such as hydroxysethyl ester or ammonia; lower alkyl amines such as methylamine, dimethylamine, ethylamine, etc. Amido.

Among the compounds represented by the general formula [1], a particularly preferred compound is, for example, (4R) -hexahydro-17,7-dimethyl-16-oxo-1,2,5- Dithiazosin-4-carboxylic acid methyl ester [Compound A], hexahydro-3,3-dimethyl-2-oxo-1,1,2,5-dithiazosin-16-methyl carboxylate [Compound B], ( 4 R) 1-hexahydrite 1-4-Rubimoyl 7, 7-dimethyl-1-2,-5-dithiazosin [Compound C], (4R)-Hexahydrid 1-7, 7 —Dimethylyl 4 -— (N, N—Dimethylcarbamoyl) -1-6-oxo-1,2,5-dithiazosin [Compound D], (4R) hexahydro-7,7 — Dimethyl-yl-6-oxo-1 1,2,5—dithiazosin-4-acetic acid acetomethyl ester [compound E] It is.

Further, the present invention is a novel compound represented by the general formula [2] or a pharmaceutically acceptable salt thereof.

Wherein the R _5, R 6 and R ₇ are identical or different and represent a hydrogen atom or a lower alkyl group, R ₈ is - CON (R ₉₎ R ₁₀ or _{- C 00 (CH 2) r} OH ( provided that 0 H R ₉ and R ₁₀ may be the same or different and represent a hydrogen atom or a lower alkyl group, p is 0 or 1, q is 1 or 2, r Represents an integer of 1 to 5, respectively. - general formula [1] represented by compounds of protecting groups arsenide mud key sheet group arsenide Doroki sheet in the lower alkyl in R _4, and in the general formula [2] R ₈ in the compound represented by at - CO 0 ( The protecting group for the hydroxy group in CH 2) r OH may be a general-purpose protecting group for a hydroxy group, and specifically, a formyl group or a lower alkanol group An alkoxy group such as a lower alkoxycarbonyl group or a phenyl lower alkoxycarbonyl group; an aryl group, a lower alkoxy lower alkyl group, or a substituted lower alkyl carbonyl group such as a halogeno lower alkanol group or a benzoyl group; Substituted alkyl derivatives such as lower alkoxy lower alkyl groups, phenyl lower alkyl groups, tetrahydrobiranyl groups, tetrahydrofuranyl groups; lower alkylsilyl groups, phenylsilyl groups; Substituted Li Le groups such as Le group.

Examples of the compound represented by the general formula [2] include (4R) -hexahydro-14-carno, moyl-17,7-dimethyl-61-oxo- 1,2,5-dithiazocine [compound C], (4 R) — Hexahydro-7,7 — Dimethylyl 4 — (N, N—Dimethylcarbamoyl) 1 6 —Oxo 1 1,2,5 — Dithiazosin [Compound D], (4R) —Hexahydro-1,7,7-Dimethyl —6 —Sodium 1,2,5 —Dithiazosin_4 _Acetoxymethyl carboxylate [Compound E] and the like. In the compounds of the present invention represented by the general formulas [1] and [2], the lower alkyl group or the lower alkanol group is a straight-chain or branched alkyl group or an alkanol having 1 to 6 carbon atoms. Refers to an aryl group.

The compounds represented by the general formulas [1] and [2] have optical isomers and, in some cases, diastereoisomers, and those containing these as an active ingredient are also included in the present invention. Included You. In addition, the present compound may be in the form of a solvate, for example, a hydrate.

The salt of the compound represented by the general formula [1] or the general formula [2] may be any salt that is acceptable as a medicament, such as a sodium salt, a calcium salt, a calcium salt, and a magnesium salt. Salts with alkali metals or alkaline earth metals, such as salts, and salts with organic amines, such as ammonium salts, methylamine salts, and triethanolamine salts, and the like.

—As a result of studying the effect of increasing the amount of glutathione (GSH) in cultured Jurkat cells on the cyclic disulfide derivative represented by the general formula [1], these cyclic disulfide derivatives were found to be more in vitro than lipoic acid. Also show a strong GSH increasing effect, and may be an excellent antioxidant. The details will be specifically described in the section on pharmacological tests below.

The present invention also provides a method for treating a disease caused by oxidative stress, which comprises administering to a patient a therapeutically effective amount of a compound represented by the general formula [1] or a pharmaceutically acceptable salt thereof. About the method.

The antioxidant of the present invention includes cataracts, ischemic diseases such as myocardial infarction and cerebral infarction, lung diseases such as emphysema and asthma, nervous system diseases such as Alzheimer's syndrome and Parkinson's disease, diabetes, and viral It is expected to be useful as a therapeutic agent for diseases involving oxidative stress such as diseases.

—The cyclic disulfide derivative represented by the general formula [1] can be administered orally or parenterally. Tablets, capsules, powders, granules, and percutaneous absorption Preparations, injections, eye drops, eye ointments, suppositories, etc., and can be formulated using commonly used techniques. The dose of the cyclic disulfide derivative represented by the general formula [1] can be appropriately selected depending on symptoms, age, dosage form, and the like. For example, when used as an oral preparation, it is usually 0.1 to 5 per day. It may be administered in a dose of 1000 mg, preferably 1-100 mg, in single or divided doses.

The results of the pharmacological tests are shown below, which are for better understanding of the present invention and do not limit the scope of the present invention. Best mode for carrying out the invention

[Production example]

A production example of the cyclic disulfide derivative represented by the general formula [2] is shown below.

Preparation Example 1 (4R) -Hexahydro-4,1-Rubamoyl-1,7,7-Dimethyl-16-Oxo-1,2,5—Dithiazosin [Compound C]

(4R) 1-hexahydro-1,7,7-dimethyl-1-6-oxo—1,2,5—dithiazosin-4—carboxylic acid 5.0 g (21.3 mm 0 1) and N—hydroxy To a solution of 3.7 g (31.8 mmo1) of droxysk cinimide in 15 ml of dimethylformamide (DMF) was added 6.6 g (31.8 g) of dicyclohexylcarbodiimide. (mmol) in DMF l O ml. Then, after cooling with ice water for 5 minutes, a catalytic amount of 4-dimethylaminopropylin was added, and the mixture was stirred at room temperature for 3 hours. A solution of 1.3 g (10.6 mmol) of oxalic acid in ethyl acetate was slowly added dropwise to the solution while taking care not to foam. The precipitated crystals were removed by filtration, the filtrate was washed with water, and the organic layer was dried over anhydrous sodium sulfate. The organic layer is concentrated under reduced pressure, The crystals obtained are washed with benzene and then with ethyl acetate, and (4R) -hexahydro-1,7,7-dimethyl-6-oxo-1,1,

2,5-Dithiazosin-4-carboxylic acid N-hydroxysuccinimide ester (5.0 g, yield 71%) was obtained.

IR (KBr): 3412, 1818, 1782, 1737, 1719, 1686, 1514, 1510, 1364, 1200, 1131, 1086, 1062, 652 cm " ¹

m 192.2-200.2 ° C

[a] _D ²⁰ -16.5 ° (DMSO, c 1.1) Then, (4R) -hexahydro-17,7-dimethyl-16-oxo-1,2,5-dithiazosin-14-carboxylic acid N- To a solution of 2.0 g (6.0 mmo1) of hydroxysuccinimide ester dissolved in 60 ml of tetrahydrofuran (THF) was added 0.83 g (6%) of 28% ammonia water. (6 mmol) in 10 ml of THF was added dropwise with stirring at room temperature. After stirring at room temperature for 30 minutes and evaporating the solvent under reduced pressure, the obtained residue was diluted with 70 ml of ethyl acetate, diluted with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogencarbonate, and then with saturated saline. Washed. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The resulting crude crystals were recrystallized with a mixed solvent of acetic acid-ethyl-n-hexane (1: 1) to obtain the target compound [Compound C].

38 g (yield 27%) of crystals were obtained.

IS (KBr): 3444, 3352, 1686, 1667, 1519, 1388, 729 cm- ¹ mp 177.0-178.0 ° C

[a] _D ²⁰ -39.8 ° (MeOH, c 1.0) Production Example 2 (4R) -hexahydro-1,7,7-dimethyl-14- (N, N-dimethylcarbamoyl) -16-oxo-1,2 , 5 —Synthesis of dithiazosin [Compound D]

(4 R) 1-hexahydro 7, 7 — dimethyl 6-oxo — 1, 2, 5 — dithiazosin 4 4 monorubonic acid N — hydroxyxsuccinimide 1.0 g (3.O mmol) was dissolved in 30 ml of THF, and 0.37 g (3.3 mmol) of 40% aqueous dimethylamine solution was added to 5 ml of THF.The solution was stirred at room temperature. While dripping. The same operation as in Production Example 1 was performed to obtain 0.36 g (yield 46%) of crystals of the target compound [Compound D].

I (KBr): 3348, 1667, 1643, 1627, 1512, 1472, 1400, 668 cm— ¹ m 159.7-160.6 ° C

[a] _D ²⁰ -61. Γ (D SO, c 0.99) Production Example 3 (4R) -Hexahydro-17,7-Dimethyl-1-6-oxo-1,2,5—Dithiazosin_4— Synthesis of Acetoxymethyl Carboxylates [Compound E]

(4R) -hexahydro-17,7-dimethyl-1-6-oxo-1,2,5-dithiazosin-4-carboxylic acid 13.0 g (55.2 mm 0 1) dimethylforma After adding 110 ml of the mixture, the mixture was stirred at room temperature. Then, 10.0 ml (71.8 mm 01) of triethylamine was added, and then methyl acetate bromide 7.O ml (71.8 mmol) was added. After stirring for 1 hour, the reaction solution was diluted with 100 ml of ethyl acetate, washed twice with 200 ml of water, and then with 50 ml of saturated saline. The obtained organic layer was dried over sodium sulfate and concentrated under reduced pressure. The resulting white solid was washed with cyclohexane to obtain 12.2 g (yield: 72%) of crude crystals. By recrystallizing this crude crystal from 2-propanol 120 m 1, the target compound was obtained. [Compound E] 11.8 g (yield 69%, recrystallization yield 97%) was obtained as colorless crystals.

IR (KBr): 3361, 2982, 1759, 1670, 1511 cm— ¹

m 136.0-137.5 ° C (i-PrOH)

[a] _D ²⁰ -107.0 ° (MeOH, c 1.0)

[Formulation example]

Formulation examples of the compounds (Compounds A to E) which are the active ingredients of the present invention are shown below.

Compound A: (4R) -hexahydro-1,7,7-dimethyl-1-6-oxo-1,2,5—dithiazosin-14-methyl carboxylate

Compound B: Hexahydro-1,3,3-dimethyl-4-oxo-1,2,5—dithiazosin-6—methyl carbonyl ester

Compound C: (4 R)-Hexahydro Draw 4-Levamoyl-7, 7-Dimethylyl 6-Oxo-1, 2, 5-Dithiazosin Compound D: (4 R)-Hexahydro-7, 7-Dimethyl Chiro 4-(N, N-dimethyl carbamoyl) 1-6-oxo-1, 2, 5--dithiazo.

Compound E: (4R) -hexahydro-1,7,7-dimethyl-1-6-oxo-1,2,5—dithiazosin-4 monoacetic acid acetonoxymethyl ester Formulation Example 1

Compound A or Compound C and a excipient were mixed, and the mixture was directly compression molded to prepare the following tablets. Compound A 0 mg Crystalline cellulose 20 mg Lactose 4 4 mg Low-substituted hydroxypropylcellulose 5 mg Magnesium stearate 1 mg

70 mg Compound C 80 mg Crystallose mouth 20 mg Lactose 44 mg Low-substituted hydroxypropylcell mouth 5 mg Magnesium stearate 1 mg

150 mg Formulation example 2 Granules

Compound B, lactose and potato starch were mixed, and a hydroxypropylcellulose solution was added as a binder to this mixture, and the following condyle-granules were prepared by a conventional method.

Compound B D 0 mg Lactose 60 mg Non-starch starch 20 mg Low-substituted hydroxypropylcellulose 4 mg talc

135 mg Compound E, mannitol and corn starch are mixed, and povidone solution is added as a binder to this mixture, and the whole is granulated in a conventional manner. Coating agent (trade name: o (Idragit RL) was coated by a conventional method to prepare the following coated granules.

Compound E 30 mg mannitol 40 mg corn starch 10 mg povidone 3 mg oil drag R L15 mg mg triacetylene 2 mg

100 mg Formulation Example 3 Capsules

The following capsules were prepared using Compound D, lactose and magnesium stearate.

Compound D 5 mg Magnesium stearate 3 mg Lactose 1 42 mg

150 mg By changing the compounding ratio of compound D to lactose, the amount of compound is JL 0 mgZ capsule, SO nig Z capsule, 50 mg Z capsule, 100 mg Z capsule Agent can be prepared.

[Pharmacological test]

For the compounds A to D, the following pharmacological tests were performed to examine the amount of daltathione (GSH) increase in the cells.

Each test drug and comparative drug of Compounds A to D is dissolved or suspended in ethanol, respectively, and their concentration is adjusted to 100 μ μ.

Two Was adjusted. Lipoic acid was used as a comparison drug, and ethanol was used as a control. The obtained ethanol solution of the test drug and the comparative drug was mixed and dissolved with an RPMI-164 (10% FBS) medium, and sterilized with a filter. Then, after the these drug solutions were aliquoted 7 5 0 ml of medium 2 4-hole plates min, Jurkat cells (approximately ^{2 xl 0 6 cells / ml in} RPMI - 1 6 4 0 (1 0% FBS) medium) were seeded each 7 5 0 1, and cultured 3 7 ° C, 24 h C 0 ₂ b incubator. 1 ml of the cell suspension was collected, centrifuged (100 rpm, 100 min), and the culture supernatant was removed. Cell sediment was added with 3% trichloroacetic acid and stored frozen at -80 ° C. This was thawed, centrifuged (1500 rpm, 5 min), and the supernatant was used for quantitative analysis of GSH. In the quantitative analysis of GSH, analysis was performed using HPLC, and the amount of GSH in the cell suspension lm 1 was calculated from a calibration curve prepared using a GSH standard. Using the remaining cell suspension, the number of cells in 1 ml of cell suspension was calculated by the trypan-blue exclusion method.

Table 1 shows the intracellular GSH after culture using each test drug and comparative drug. The values in the table are the average values of 2 to 4 cases.

Three Drug GSH picture 1/1 0 ⁶ cells Compound A 9. 3 is apparent from compound B 8. 4 Compound c 1 1. 3 Compound D 1 2.5 lipoic acid 6.6 Con Bok port one Honoré 2.5 Table 1 As described above, all of the compounds A to D have a stronger action of increasing daltathione (GSH) than lipoic acid. Therefore, the antioxidant containing the cyclic disulfide derivative of the present invention as an active ingredient is useful for oxidizing a living body. It affects the process of dysfunction, such as ischemic diseases such as myocardial infarction and cerebral infarction, lung diseases such as emphysema and asthma, nervous system diseases such as Alheimer's syndrome, Parkinson's disease, cataract, diabetes, and viral diseases. Useful as a therapeutic agent. Industrial applicability

TECHNICAL FIELD The present invention relates to an antioxidant containing a cyclic disulfide derivative as an active ingredient. According to the present invention, it acts on the oxidative damage process of the living body, and ischemic diseases such as myocardial infarction and cerebral infarction, lung diseases such as emphysema and asthma, nervous system diseases such as Alzheimer's syndrome and Parkinson's disease, cataract, and diabetes Thus, an antioxidant effective for treatment of viral diseases and the like is provided.

Four

Claims

The scope of the claims

1. An antioxidant comprising a compound represented by the general formula [1] or a pharmaceutically acceptable salt thereof as an active ingredient.

Wherein R i, R 2 and R ₃ are the same or different and are a hydrogen atom or a lower alkyl group, R ₄ is a carboxylic acid or its ester or amide, m is 0 or 1, n Represents 1 or 2, respectively.

2. A method for treating a disease caused by oxidative stress, which comprises administering to a patient a therapeutically effective amount of a compound represented by the general formula [1] or a pharmaceutically acceptable salt thereof.

Where R! , R 2 and R ₃ are the same or different and represent a hydrogen atom or a lower alkyl group, R ₄ is a carboxylic acid or its ester or amide, m is 0 or 1, and n is 1 or 2 Are respectively shown.

3. Use of a compound represented by the general formula [1] or a pharmaceutically acceptable salt thereof for the manufacture of an antioxidant.

4. A compound represented by the general formula [2] or a pharmaceutically acceptable salt thereof.

Wherein the R _5, R 6 and R ₇ are identical or different and represent a hydrogen atom or a lower alkyl group, R ₈ is - CON (R 9) R ₁₀ or _{_{- COO (CH 2) r OH}} ( provided that 〇 H group May be protected by a protecting group), R ₉ and R ₁₀ are the same or different and are a hydrogen atom or a lower alkyl group, p is 0 or 1, Q is 1 or 2, and r is An integer of 1 to 5 is shown.

6