JPH10338690A - Ascorbic acid derivative and its use - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject new compound capable of selectively inducing apoptosis to unnecessary cells or pathogen cells and useful for prevention or treatment of diseases generated from the cells. SOLUTION: This compound is represented by formula I (X, Y and Z are each H, a alkyl halide, OH or an alkoxy; L is H or deuterium; R is H, an alkyl, an alkoxyalkyl or an acyl), e.g. 5,6-O-p-trifluoromethylbenzylidene-ascorbic acid. The compound of formula I (R is H) is obtained by reacting a compound of formula II with ascorbic acid of formula III in the presence of a catalyst such as p-toluenesulfonic acid in a dipolar solvent such as dimethylformamide. The compound of formula I has low toxicity and is useful as a carcinostatic agent and an anticancer agent and further useful for prevention and treatment of infectious diseases due to HIV virus, especially AIDS.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to an ascorbic acid derivative represented by the following formula [I] or a pharmaceutically acceptable salt thereof, and more specifically to selectively induce apoptosis of unnecessary or pathogenic cells, The present invention relates to a novel compound useful for prevention and / or treatment of a disease caused by the cell, and use thereof.

[0002]

2. Description of the Related Art In recent years, with respect to death of cell tissue, apoptosis (apoptosis) means cell suicide or suicide.
Apoptosis). This apoptosis, unlike necrosis, which is a pathological cell death, is considered to be cell death based on the expression of the gene of the cell integrated from the beginning. That is, some external or internal factors trigger the activation of genes that carry out apoptosis, apoptosis proteins are biosynthesized based on these genes, and the cells themselves actively self-generate by the generated apoptosis proteins. It is thought to break and die.

[0003]

An object of the present invention is to provide a novel compound having a selective apoptosis-inducing effect on unnecessary or pathogenic cells. Another object of the present invention is to provide a pharmaceutical use of the novel compound.

[0004]

The present inventors have conducted intensive studies to achieve the above object, and as a result, have found that ascorbic acid derivatives represented by the following general formula [I] or pharmaceutically acceptable salts thereof can be obtained. The present inventors have found that the compound has an apoptosis-inducing effect on unnecessary or pathogenic cells and is useful for prevention and / or treatment of diseases caused by the cells, and completed the present invention.

That is, the present invention provides a compound represented by the general formula [I]:

[0006]

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(Wherein X, Y and Z may be the same or different and each represents hydrogen, alkyl halide, hydroxyl or alkoxy, L is hydrogen or deuterium, R is hydrogen, alkyl, alkoxyalkyl or acyl The present invention relates to an ascorbic acid derivative represented by the following general formula (I) or a pharmaceutically acceptable salt thereof, and an ascorbic acid derivative represented by the general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient; For selectively inducing apoptosis of the cells to prevent and / or treat diseases caused by the cells.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION In the present specification, examples of the halogen in the alkyl halide include fluorine, chlorine, bromine and iodine, and fluorine is preferred. The alkyl of the halogenated alkyl may be linear or branched, and is preferably a lower alkyl having 1 to 6 carbon atoms. Specifically, methyl, ethyl, propyl, isopropyl, butyl,
Examples include isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl and the like. More preferably, it is a lower alkyl having 1 to 4 carbon atoms. Examples of the halogenated alkyl include those in which some of the hydrogen atoms of the aforementioned alkyl are halogenated, for example, (C
_{F 3) 2 CHCH 2 -,} CF 3 CH 2 -, CHF 2 -,
CH ₂ F— or the like, or one in which all hydrogen atoms are halogenated, for example, trifluoromethyl, pentafluoroethyl, or the like may be used.

In the present specification, the alkoxy is preferably a lower alkoxy having 1 to 6 carbon atoms, specifically, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentoxy. , Isopentyloxy, isopentyloxy, neopentyloxy, hexyloxy and the like.

As the alkyl in the present specification, those exemplified as the alkyl of the above-mentioned alkyl halide can be mentioned. As the alkoxyalkyl, the total number of carbon atoms is 3
To 7 are preferred, and specific examples thereof include methoxyethyl, ethoxyethyl, propoxyethyl, isopropoxyethyl, butoxyethyl, 2-methoxy-1-methylethyl, 2-ethoxy-1-methylethyl and the like.
Acyl may be either aliphatic acyl or aromatic acyl. Examples of the aliphatic acyl include those having 2 to 20 carbon atoms, and specifically, acetyl, propanoyl, butanoyl, 2-isobutanoyl, pivaloyl, hexanoyl,
Examples include undecanoyl, octanoyl, 5,7,7-trimethyl-2- (1,3,3-trimethylbutyl) -octanoyl, palmitoyl, cyclohexanoyl and the like. Examples of the aromatic acyl include henzoyl and the like.

The reactive substituent in the present specification includes, for example, halogen, and examples of the halogen include those described above as the halogen of the alkyl halide.

The compound of the present invention is specifically represented by the substituents X, Y, Z, L shown in Table 1 in the formula [I].
And compounds having R.

[0013]

[Table 1]

The ascorbic acid derivative [I '] of the present invention wherein R is hydrogen is a compound represented by the formula [II]:
I]] to produce ascorbic acid.

[0015]

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Wherein X, Y, Z and L are as defined above.

This reaction is preferably carried out using a catalyst usually used in known reactions for producing acetal from aldehydes, such as the reaction between benzaldehyde and dialcohol or polyhydric alcohol.
Examples of the catalyst include an acidic catalyst such as sulfuric acid and p-toluenesulfonic acid.

The reaction is preferably carried out in a dipolar solvent such as dimethylformamide, dimethylsulfoxide, dimethylacetamide and the like.

The reaction conditions and the solvent used are appropriately selected depending on the reactivity and solubility of the reactants.

Also, by a known method for preparing acetal from aldehyde, such as reaction of substituted benzaldehyde or lower acetal thereof with dialcohol or polyhydric alcohol in the presence of an acidic catalyst. Can also be prepared.

When R is alkyl, alkoxyalkyl or acyl, for example, the following synthesis method can be used.

First, there is a method for synthesizing the compound [I "] of the present invention by reacting the compound [I '] with the compound [IV].

[0023]

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Wherein X, Y, Z and L are as defined above, R ′ is alkyl, alkoxyalkyl or acyl, and X is a reactive substituent.

This is a normal de-HX reaction. The reaction is performed in an inert organic solvent such as methylene chloride, chloroform and the like. The reaction is preferably performed in the presence of a catalyst such as a base (for example, a tertiary amine such as pyridine and triethylamine).

Second, 3-O-substituted ascorbic acid [II
I ′] is reacted with the compound (II) to give the compound of the present invention (I ″
] Is synthesized.

[0027]

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(Wherein X, Y, Z, L and R ′ are as defined above) This synthesis can be carried out according to the above-mentioned first reaction.

The compound [III '] is known and disclosed in JP-A-58-131978 and JP-A-1-313476.
No. 6,009,036.

The salts of the ascorbic acid derivative of the present invention include, for example, alkali metal salts such as lithium, sodium and potassium; alkaline earth metal salts such as calcium and magnesium; aluminum salts; triethylamine, basic amino acids (lysine, arginine, Pharmaceutically acceptable salts such as organic salts such as histidine).

The compound of the present invention has low toxicity and has an apoptosis-inducing action on unnecessary or pathogenic cells, and thus is useful for the prevention and / or treatment of diseases caused by the cells. For example, it is useful as an anticancer agent, an anticancer agent, and also for the prevention and / or treatment of many diseases caused by various viral infections, infection by HIV virus, particularly acquired immunodeficiency syndrome (AIDS).

When the compound of the present invention is used as a prophylactic / therapeutic agent for a disease, the compound of the present invention is appropriately mixed with pharmacologically acceptable additives (eg, carriers, excipients, etc.), auxiliary components, etc. , Granules, tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions, injections, syrups,
A pharmaceutical composition in the form of an external preparation or the like can be administered orally, parenterally or externally.

Such auxiliary ingredients include, for example, solid, semi-solid or semi-solid such as water, glucose, lactose, gelatin, mannitol, starch paste, magnesium trisilicate, corn starch, keratin, colloidal silica, potato starch, urea and the like. Includes those that can be used effectively for the production of liquid formulations. In addition, auxiliary substances such as stabilizers, fillers, colorants and fragrances can be included. Preservatives can also be included to maintain the activity of the compound of the present invention, and contain a sufficient amount of the compound of the present invention to produce the desired therapeutic effect on the progress or condition of the associated disease. Should be.

When administering the agent for preventing or treating a disease of the present invention to a human, an effective amount of the compound of the present invention is desirably administered parenterally, for example, by injection, or orally. The dose of the compound of the present invention varies depending on sex, age, body weight, symptom, therapeutic effect, administration method, administration period and the like, and for example, about 1 to 3000 mg / day for an adult patient.

[0035]

EXAMPLES The present invention will be described in more detail with reference to Examples, Experimental Examples and Formulation Examples, but the present invention is not limited thereto.

Example 1 Synthesis of 5,6-O-p-trifluoromethylbenzylidene-ascorbic acid (compound 1) (Part 1) In 150 ml of a benzene solution containing 20 g of ascorbic acid and 20 g of p-trifluoromethylbenzaldehyde, p
-70 mg of toluenesulfonic acid was added, and the mixture was heated under reflux for 2 hours and simultaneously azeotropically dehydrated. After completion of the reaction, the resultant was washed with a sodium hydrogen carbonate solution and dried over anhydrous sodium sulfate. After evaporating the solvent, the target compound was separated and purified by gel filtration chromatography.

Example 2 Synthesis of Compound 1 (Part 2) Ascorbic acid (20 g), p-trifluoromethyl-dimethoxytoluene (20 g) and a catalyst amount (160 mg) of p
-Toluenesulfonic acid is converted to dimethylformamide 60m
and heated under reflux for 4 hours. At the same time, methanol produced by azeotropic distillation was removed. After completion of the reaction, petroleum ether
The crystals were washed with an aqueous system and then crystallized from a hexane-water system to obtain the target compound.

Example 3 Synthesis of Compound 1 (Part 3) Ascorbic acid (20 g) and p-trifluoromethylbenzaldehyde (20 g) were mixed with dimethylformamide 150
Dissolved in ml. 4 ml of concentrated sulfuric acid was added and reacted at room temperature for 2 days. The solvent was distilled off to obtain the target compound.

Example 4: Synthesis of sodium 5,6-Op-trifluoromethylbenzylidene-ascorbate (Compound 2) Compound 1 (15 g) was suspended in 300 ml of water, and 5 g of sodium hydrogencarbonate was added. Lyophilize the solution,
The desired compound was obtained. The physical property data were as follows.

HPLC: C ₁₈ -ODS (octadecylsilyl) type reversed phase column, developing solvent is methanol-water (9:
In 1), it was a single peak. IR (cm ^-1 , KBr): 830, 1100, 1320, 1570, 172
0, 3400 ¹ H-NMR [ppm, solvent is D ₂ O, target is DSS
(Sodium 2,2-dimethyl-2-silapentane-5-sulfonate)]: 4.0-4.5 (m, 2H), 4.5-4.9 (m, 4
H), 5.9 (m, 1H), 7.3-7.9 (m, 4H)

Example 5: 3-On-ethyl-5,6-
Synthesis of Op-trifluoromethylbenzylidene-ascorbic acid (Compound 3) 5,6-Op-trifluoromethylbenzylidene-ascorbic acid 15 g, sodium methoxide 3 g, ethyl iodide 10 g, and DMSO (dimethyl sulfoxide) 150 ml Was prepared and reacted at room temperature for 72 hours. The reaction solution was extracted with ethyl acetate, and the extract was washed with a saturated aqueous solution of sodium chloride. The ethyl acetate layer was separated, dried over anhydrous sodium sulfate, and the solvent was distilled off. The target compound was isolated by twice TLC silica chromatography (developing solvent: first time, methanol-toluene-ethyl acetate system, second time, ethyl acetate-toluene system).
The physical property data were as follows.

HPLC: C ₁₈ -ODS reverse phase column, developing solvent was a single peak in methanol-water (9: 1). IR (cm ^-1 , NEAT): 820, 1090, 1320, 1370, 1680,
1720, 3000, 3300 ¹ H-NMR [ppm, solvent: CDCl ₃ + d ₄ -methanol, target: TMS (tetramethylsilane)]: 1.3
(t, 3H), 3.7-4.8 (m, 7H), 5.7-6.0 (m, 1H), 7.1-8.0
(m, 4H)

Example 6: 3-O-palmitoyl-5,6
Synthesis of -Op-trifluoromethylbenzylidene-ascorbic acid (compound 4) (part 1) A target compound was prepared according to Example 1 except that 3-O-palmitoyl-ascorbic acid was used instead of ascorbic acid. .

Example 7 Synthesis of Compound 4 (Part 2) 8 ml of pyridine was added to 100 ml of a methylene chloride solution containing 3 g of 5,6-Op-trifluoromethylbenzylidene-ascorbic acid, and 3.3 g of palmitoyl chloride was added.
10 ml of a methylene chloride solution was added dropwise. After reacting at room temperature for 3 hours, the mixture was washed successively with dilute hydrochloric acid and water, dried over anhydrous sodium sulfate, and the solvent was distilled off. The residue was separated by gel chromatography to obtain the target compound. Physical properties data of compound 4 were as follows.

HPLC: C ₁₈ -ODS reverse phase column, developing solvent was a single peak in methanol-water (1: 9). IR (cm ^-1 , KBr): 820, 1120, 1320, 1680, 174
0, 2800, 2900, 3400 ¹ H-NMR [ppm, solvent: CDCl ₃ + d ₄ -methanol, target: TMS]: 0.6-1.8 (m, 31H), 3.5-4.8
(m, 5H), 5.6-5.9 (m, 1H), 7.3-7.5 (m, 4H)

Experimental Example 1 The apoptosis-inducing effect of Compound 2 on human promyelocytic leukemia cells (HL-60) was confirmed. HL-
After treating 60 cells (5 × 10 ⁵ cells) in the presence of the compound of the present invention at 37 ° C. for 5 hours, the occurrence of DNA fragmentation characteristic of apoptosis was confirmed by agarose gel electrophoresis. (Apoptosis induction ability) was calculated. Table 2 shows the results.

[0047]

[Table 2]

Experimental Example 2 In accordance with Experimental Example 1, the apoptosis-inducing effect of each of the compounds 2 to 4 was confirmed. The final concentrations of Compounds 2 to 4 were 3 mM, 1 mM, and 0.1 mM, respectively. Table 3 shows the ratio of the fragment compounds of DNA (induction of apoptosis).

[0049]

[Table 3]

Experimental Example 3 (Toxicity) When a solution of compound 2 (10 mg / ml) was administered to mice, no mortality was observed even when intravenously administered at 1000 mg / kg body weight.

Formulation Example 1 (tablets) (1) Compound 2 50 g (2) Fine particles for direct hit No. 209 (manufactured by Fuji Chemical Co., Ltd.) 70 g Magnesium aluminate metasilicate 20% Maize starch 30% Lactose 50% (3) Crystal Cellulose 60g (4) CMC calcium 18g (5) Magnesium stearate 2g

After uniformly mixing (1) to (4), (5) was added and further mixed, and the mixed powder was tabletted to give a tablet of 200 mg per tablet. The tablet may be coated, if necessary, with a commonly used enteric or gastric film coating (eg, polyvinyl acetal diethylaminoacetate) or an edible colorant.

Formulation Example 2 (Capsule) (1) Compound 2 1000 g (2) Lactose 960 g (3) Magnesium stearate 40 g

The above components were uniformly mixed, and the mixed powder was filled into hard gelatin capsules at 200 mg each.

Formulation Example 3 (Injection) (1) Compound 2 100 mg (2) Glucose 100 mg (3) Water for injection 10 ml in total amount

The solution obtained by dissolving (1) and (2) in (3) was filtered through a membrane filter and then sterilized and filtered again.
The filtrate was aseptically dispensed into vials, filled with nitrogen gas, and sealed to give an intravenous injection.

[0057]

EFFECTS OF THE INVENTION The ascorbic acid derivative of the present invention or a pharmaceutically acceptable salt thereof has low toxicity, and has an apoptosis-inducing effect on insoluble or pathogenic cells. Prevention and / or
Or it is useful for treatment. For example, it is useful as an anticancer agent, an anticancer agent, and also for the prevention and / or treatment of many diseases caused by various viral infections, infection by HIV virus, particularly acquired immunodeficiency syndrome (AIDS).

Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 31/365 AGZ A61K 31/365 AGZ

Claims (2)

[Claims] 1. A compound of the general formula [I]: (Wherein X, Y and Z may be the same or different and each represents hydrogen, alkyl halide, hydroxyl group or alkoxy, L represents hydrogen or deuterium, R represents hydrogen, alkyl, alkoxyalkyl or acyl, respectively) Or a pharmaceutically acceptable salt thereof. 2. An ascorbic acid derivative or a pharmaceutically acceptable salt thereof according to claim 1 as an active ingredient, which selectively induces apoptosis of unnecessary or pathogenic cells to prevent diseases caused by the cells. And / or an agent for treating.