NL8101630A - 3,7,11,15-TETRAMETHYL-2,4,6,10,14-HEXADAPAPENTOIC ACID. - Google Patents

Description

s * *

Br / B1 / 1h / 18 3,7,11,15-tetramethyl-2,4,6,10,14-hexadecapentaenoic acid.

The invention relates to a new compound, namely the 3,7,11,15-tetramethyl-2,4,4,10,10,14-hexadecapentaenoic acid of the formula 1 as well as pharmaceutically acceptable salts thereof. The invention further relates to methods of preparation thereof, an anticancer agent containing the acid or a pharmaceutically acceptable salt thereof, and to a therapeutic composition for the treatment of keratinization associated with skin diseases.

10 W. Bollag et al. Iri Europ. J. CJancer, vol. 10, p. 731 {1974}, that retinoids such as the ethyl ester of 9- (2,3,6-trimethyl-4-methoxyphenyl) -3,7-dimethyl-2,4,6 , 8-nonatetraenoic acid are effective against cancer. However, these retinoid compounds are highly toxic and present further problems such as causing vitamin A hypervitaminosis administration.

The novel compound of the formula I provided according to the invention exhibits anti-cancer activity, causes practically no hypervitaminosis of vitamin A 20 and is also little toxic in other respects.

The compound of the invention can. be prepared according to the following procedures.

Method A

This process includes: (1) Reaction of a compound of formula II with a Wittig reagent prepared from a compound of general formula 3 wherein X represents a halogen atom and a lower alkyl group to form a compound of the general formula 4, which has the meaning as defined above, and (2) hydrolysis of the compound of the general "formula 4" thus obtained in the presence of a base for preparing the compound of the formula 1.

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Examples of the Wittig reagents used in the above-described step (1) and prepared from a compound of the general formula 3 are phosphorus compounds, which are prepared by reacting the compound of the general formula 3 and triphenylphosphine, phenyl dialkoxyphosphine, trialkylphosphite and the like. The preparation of the reagent and the Wittig reaction using the reagent are carried out by conventional methods such as the method described by Wadworth et al., 10 in J. Am. Chem . Soc., Vol. 83, p. 1733 (1966), the method described by Greenwald et al., In J. Örg. Chem., Vol. 28, p. 1128 (19631 and the method described by Horner et al, in Ber. Vol. 95, p. 581 (19621.

In the above step (21), the hydrolysis is carried out in the presence of a base, which is generally used for the hydrolysis of carboxylic acid esters, such as sodium hydradide and potassium hydroxide.

Method B—

The method comprises: 20 (1) reacting a compound of formula 5 with a Wittig reagent prepared from a compound of general formula 6, wherein X represents a halogen atom and a lower alkyl group, to form a compound of the general formula 4, and hydrolysis of the compound of the general formula 4 obtained in the presence of a base to prepare the compound of the formula 1.

Each of the above-described steps (Ij.eri (21) can be performed in the same manner as described for Method A.

Method C

This process comprises: (11 reaction of a compound of the general formula 7 wherein Y represents a lower alkyl group or an aryl group, with a compound of the general

Formula 6 to form a compound of the general Formula 8, wherein Y has the above meaning, and

> * -3- (2) subjecting the compound of the general formula 8 thus obtained to desulphination and hydrolysis of the ester in the presence of a base to prepare the compound of the formula 1.

Step (1) is performed in the presence of a base. Examples of the bases are n-butyl lithium and phenyl lithium. Examples of the reaction solvents are tetrahydrofuran, diethyl ether and 1,2-dimethoxyethane.

The conversion is generally carried out at a temperature below room temperature.

The step (2) can be performed in the same manner as the step (2) of the above-described method (A).

Examples of the substituents which may be present in the general formulas 3,4,6,7 and 8 are the following: halogen atoms, such as chlorine, bromine and iodine atoms as substituent X; lower alkyl groups, such as methyl, ethyl and propyl groups as substituent R1, and lower alkyl groups, such as methyl, ethyl and propyl groups, and aryl groups, such as phenyl and p-tolyl groups as substituent Y.

Examples of the pharmaceutically acceptable salts of the compound of the formula 1 are the sodium and the potassium salt thereof.

The novel compound of the formula 1 provided according to the invention also shows therapeutic activity in the treatment of skin diseases, which are associated with keratinization.

Examples of the skin diseases associated with keratinization which can be treated with the compound of the formula J. are skin diseases with symptoms, such as hyperkeratosis, parakeratosis and dyskeratosis. More particularly, examples of these skin diseases are psoriasis, acne, acne vulgaris, Darier's disease, palmoplantar pustulossis, lichen planus, ichthyosis, erythroderma, pityriasis rubra pilasis and keratosis senilis.

For the treatment of keratinization-associated skin diseases, steroid-type preparations for external use are used. However, these preparations have strong side effects, so they do not have 8101630 φ ΐ

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«-4- are useful for repeated administration over a long period of time and the treatment by administration of a large amount of the composition.

In contrast, the 3,7,11,15-5 tetramethyl-2,4,6,10,14-hexadecapentaenoic acid of the invention has the action of inhibiting skin keratinization and exhibiting low toxicity.

The results of pharmacological tests and on the toxicity of the Compound of the invention are described below.

Pharmacological tests (anti-cancer activity) (11 Test instruction.

Mice (ICR, female, 60 days old) were shaved (5 cm) at the back of the neck. 7.12-15 dimethylbenzo-27-anthracene was dissolved in acetone to a solution at a concentration of 75 mg / 100 ml. The solution thus prepared was applied to the mice in an amount of 0.2 ml per mouse on the day they were 60 days old and on the day they were 75 days old.

Croton oil was dissolved in acetone to a solution at a concentration of 250 mg per 100 ml, which solution thus prepared was applied to the mice in an amount of 0.2 ml per mouse twice a week. t until the start of treatment. Then itself. 3-7 papillomata 25 (each with a diameter of 3-8 mm and a total diameter of 30-60 mml per mouse had developed was started with the treatment.

The test compound was dissolved in groundnut oil "to a solution at a concentration of 20 mg / ml and administered orally to the mouse. The solution was administered 10 times over 14 days (once per day), after which the diameters of the papillomata were measured on the 14th day to determine the total diameter per mouse.

(21 Trial connection.

3.7, 1.15-tetramethyl-2,4,6,10,14-hexadecapentaenoic acid (the compound of the invention).

Ethyl ester of 9- (2,3,6-trimethyl-4-methoxy-phenyl) -3,7-dimethyl-2,4,6,8-nonatetraenoic acid (comparative 8101630 δ-5 compound).

(3) The results are shown in Table A below.

TABLE A

Test Number Papilloma (total diameter / binding mice -

Average value Average value Relative (Oth day) (14th day) increase or. . decrease in nut oil 3 33.9 “39.7 m, + 17.1%

Compound of the invention (200 mg / kg. 5 37.5 mm 21.3 mm - 43.2% day)

Comparative compound 3 58/1 mm 32.7 mm -43.7% (40 mg / kg.day)

As shown in Table A above, the compound of the invention is effective against the papilloma.

Pharmacological tests (inhibition of keratinization).

(1) Test prescription.

In a Petri dish (diameter of 6 cm), where 10 were placed in 8 watch glasses (diameter of 15 mm), 5 ml of a suspension of the abnormal epithelial cells

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from a rat bladder, designated as BES 20B (about 2 x 10 cells / ml), incubation took place for 24 hours at 37 ° C and a carbon dioxide concentration of 5%. Each of the watch glasses thus treated was placed in 2 ml of Eagle's MEM medium containing the test compound in different concentrations, after which further incubation was carried out at 37 ° C and a carbon dioxide concentration of 5%. The medium was changed at 2-3 day intervals. On the second, fifth, eighth and fourteenth days from the start of incubation, the watch glass was taken out of the medium and subjected to the Papanicolaou stain test to determine the degree of keratinization. The observation was performed by measuring 8101630 m 3-6 the absorption spectrum in the range 400-750 nm, after which the KI (keratinization index) was calculated according to the following equation.

Absorbance peak near 490 nm attributed to the keratinization cells. . .........

KI = -

Absorption peak in the vicinity of 640 nm attributed to the non-keratinized cells.

A KI value of 1.0 or more indicates strong keratinization, while a KI value of 0.5 or less indicates virtually no keratinization.

The BES 20B cells were incubated for comparison in a medium containing no test compound.

(2) Test connection.

3,7,11,15-tetramethyl-2,4,6,10,14-hexadecapentaenoic acid (the compound of the invention).

(3) Trial results.

The results are summarized in Table B.

: .TABLE · B .................

• _κι; c.: .........

Duration of incubation 2 days. 5 days . 8 days. . 1.4. days

Control 0.43 1.10 3.27 3.08

Compound according to the invention 0.1 pg / ml 0.43 0.67 0.55 0.52 1.0 pg / ml 0.42 0.46 0.38 0.39 5.0 jug / ml 0.48 0.50 - 0 , 22

In the control experiment, the KI value on the fifth day after incubation started was 20.0, indicating strong keratinization. In contrast to the control result, the results obtained at different concentrations of the compound of the invention yield KI values of less than 1.0 in all tests, indicating inhibition of keratinization.

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Toxicity tests (1) Test instruction.

The test compound was repeatedly administered to a group of 6 mice (ICR strain, female] for 14 days. The amount administered was 40 mg / kg.

5 day, 200 mg / kg.day and 400 mg / kg.day for the compound of the invention and 200 mg / kg.day for the comparative compound. In the course of administration, the weight gain or loss of the mice, the occurrence of death, etc. were observed.

10 (2) Test compound.

The compounds described in the pharmacological tests (anti-cancer activity) were used.

(3] Test result.

(a) Weight gain and loss.

The results are shown in Table C below.

(b] Mortality.

All mice treated with the comparative compound in an amount of 200 mg / kg.day had died by the eighth day, while no mortality was observed in the mice treated with the compound of the invention.

(c] Hair loss.

Hair loss was observed on the sixth day in each mouse treated with the comparative compound at 200 mg / kg.day, while no hair loss was observed in the mice treated with the compound of the invention .

(d) Cyanosis.

Cyanosis was observed on the seventh day in each of the mice treated with the comparative compound in an amount of 200 mg / kg.day, while no cyanosis was observed in the compound treated with the invention mice.

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The items in the toxicity tests, hair loss and weight change are known to indicate hypervitaminosis of vitamin A. Since hair loss and weight loss were observed to a very great extent in the group of mice treated with the comparator, assumed that hypervitaminosis of vitamin A occurred. In contrast, such problems were not observed at all in the group of mice treated with the compound of the invention.

In connection with the results of the pharmacological and toxicity tests described above, the compound of the invention is considered to be a very safe compound and is believed to be of value as an anti-cancer agent and as a therapeutic agent for treatment of skin diseases associated with keratinization.

The compound according to the invention can therefore be used for the prevention and treatment of cancer and precartinous conditions and also for the treatment of skin diseases associated with keratinization, such as acne and psoriasis vulgaris, as well as in the treatment of allergic and inflammatory skin diseases. In addition, the compound of the invention can be used to treat muscle tissue diseases caused by inflammation, degeneration or plastic surgery.

For the administration or anti-cancer agent and as a therapeutic agent for the treatment of skin diseases associated with keratinization, the compound of the invention is administered orally in the form of powders, granules, pills, hard capsules, etc., or parente - orally in the form of ointments, suppositories, indicative solutions, etc. The dosage generally requires 40 mg to 4 grams per day for an adult. When the compound of the invention is used in the form of a composition for external use, the dosage can be varied depending on the conditions of the disease.

The compound according to the invention can be combined with a generally usable carrier for medicinal use in the conventional manner for preparing the above-described preparations.

The methods of preparing the compound of the invention are illustrated by the examples below, which, however, are not intended to limit the invention.

Example I

To a suspension of 5.0 g of 55% sodium hydride (oily) in 60 ml of n-hexane was added 28.6 g of triethyl phosphonoacetate. The mixture was then heated to reflux, after which 20 g of 6, 10,14-trimethyl-3,5,9,13-pentadecatetra-one-2-one were added dropwise with stirring. After 30 minutes, the reaction liquid was poured into 200 ml of water, after which 500 ml of n-hexane were added for the extraction. The n-hexane phase was separated twice with 10 ml each of a mixture of methanol and water (2: 1). washed and pimped * · '· centered. The concentrate thus obtained was purified by column chromatography on silica gel to obtain 18 g of ethyl ester of 3,7,11,15-tetramethyl-2,4,6,10,14-hexa-decapentaenoic acid.

To 10 g of the ethyl ester of 3.7, 11,15-tetramethyl-2,4,6,1,14-25 hexadecapentaenoic acid obtained in the manner described above, a solution of 3.9 g of potassium hydroxide in 30 ml was added. isopropanol was added and the mixture was stirred at 50 ° C for one hour. The reaction liquid was then poured into ice water. adjusted to acid reaction by addition of hydrochloric acid and extracted with 100 ml diethyl ether. The ether phase was washed with water, dried over magnesium sulfate and concentrated to give 9.0 g of an oil. The oil was dissolved in 50 mL of n-hexane and crystallized at -20 ° C to give 4.0 g of 3.7,11,15-tetramethyl-2,4,6,10,14-hexadeca-35 pentaenoic acid in the form of pale yellow strokes was obtained. __

Melting point: 78.4 ° C -

Mass spectrum (m / e): 302 (M +) 8101630 Λ -11-

Infrared absorption spectrum (cm KBr tablet): 34.50, 2900, 1680, 1595 NMR spectrum (S CDCl ^): 1.61 (6H, s), 1.68 (3H, s), 1.86 (3H, s), 1.92 - 2.24 (8H, b), 2.35 (3H, s), 5.10 5 (2H, b), 5.76 (1H, bs), 5.98 (1H, d, J = 11 Hz), 6.20 (1H, d, J = 15 Hz), 6.90 (1H, dd, J = 11 Hz, 15 Hz), 11.63 (1H, b)

Ultraviolet absorption spectrum: T ^^^ anole: 304 nm.

Example II

To a suspension of 4.8 g of sodium ethanolate in 100 ml of n-hexane was added 18 g of diethyl ester of 3-ethoxy-carbonyl-2-methyl-2-propanylphosphonic acid. To the mixture were added 10 g of 3.7, 15 l of trimethyl-2,6,10-dodecatriene-1-al under stirring at room temperature. After one hour, the reaction liquid was poured into 50 ml of water and the n-hexane phase was separated. The n-hexane phase was washed twice with 50 ml each of a mixture of methanol and water (2: 1) and concentrated. The concentrate obtained in this way was purified by silica gel column chromatography to obtain 14.5 g of ethyl ester of 3,7,11,15-tetramethyl-2,4,6,10,14-hexadecapentaenoic acid in the form of yellow needles. . The product thus obtained was identified by the melting point, mass spectrum, NMR spectrum, infrared absorption spectrum and ultraviolet absorption spectrum in the same manner as described in Example 25.

Example III

10 g of 1-p-tolyl-30 sulfonyl-3,7,11-trimethyl-2,6,10-dodecatriene were dissolved in 100 ml of tetrahydrofuran and the solution was cooled to -50 ° C. To the solution were added dropwise 18.5 ml of a 15% fis n-butyl lithium n-hexane solution while stirring and under a nitrogen flow, while keeping the temperature of the solution at -50 ° C. Then, 300 ml of a tetrahydrofuran solution containing 5.7 g of ethyl ester of 4-bromo-3-methyl-2-butenoic acid were added dropwise to the solution thus obtained. After a period of 30 minutes, 100 ml of a 10% aqueous ammonium chloride solution was added and the temperature of the mixture was allowed to rise to room temperature. The mixture was then extracted twice with 200 ml of n-hexane each time.

The n-hexane phase was washed three times with 100 ml of water each time, dried over magnesium sulfate and concentrated, whereby 13 g of ethyl ester of 3.7, 1.15-tetrainethyl-5-p-tolyl-sulfonyl-2,6,10,14 hexadecatetraenoic acid were obtained.

To 10 g of the ethyl ester obtained as described above was added a solution of 4.6 g of potassium hydroxide in 50 ml of isopropanol and the mixture was stirred at 50 ° C for 3 hours. The reaction liquid was then poured into ice water, acidified by the addition of hydrochloric acid and extracted with 100 ml diethyl ether. The diethyl ether phase was washed with water, dried over magnesium sulfate and concentrated to give 6 g of an oil. The oil was dissolved in 30 ml of n-hexane and crystallized at -20 ° C to give 1.8 g of 3.7, 1.15-tetramethyl-2,4,6,10,14-20 hexadecapentaenoic acid in the form of light yellow needles were obtained.

The product obtained in this way was identified in the same manner as in Example 1, that is, from the melting point, the mass spectrum, 25 NMR spectrum, infrared absorption spectrum and ultraviolet absorption spectrum.

• Example IV

Granules 3,7,11,15-tetramethyl-2.4.6.10.14-hexadecapentaenoic acid 50 g silicic anhydride 30 g

Crystalline cellulose 50 g

Corn starch 36 g

Hydroxypropyl cellulose 10 g 22 Magnesium stearate 4 g

The above recipe was performed in a conventional manner to form granules (180 mg per granule).

8101630

Claims (6)

2. Process for preparing a compound of the formula I or a pharmaceutically acceptable salt thereof, characterized in that a compound of the formula 2 is reacted with a Wittig reagent prepared from a compound of the general formula 3, wherein X represents a halogen atom and R 1 represents a lower alkyl group, to form a compound of the general formula 4, which has the above meaning, and the compound obtained in this way in the presence of a base hydrolyzes. 3. Process for preparing a compound of the general formula 1 or a pharmaceutically acceptable salt thereof, characterized in that a compound of the formula 5 is reacted with a Wittig reagent prepared from a compound of the general formula 6, wherein X represents a halogen atom and a lower alkyl group, to form a compound of the general formula 4, wherein R 1 has the above meaning, and the compound obtained in this way in the presence of a base hydrolyzes. 4. A process for preparing a compound of the formula I or a pharmaceutically acceptable salt thereof, characterized in that a compound of the general formula 7 in which Y represents a lower alkyl group or an aryl group is reacted with a compound of the general formula 6, wherein X represents a halogen atom and R 1, a lower alkyl group, to form a compound of the general formula 8, wherein Y and R 1 have the above meaning, and the compound thus obtained subject to desulphination and hydrolysis of the ester in the presence of a base. 5. A composition characterized by 3,7,11,15-tetramethyl-2,4,6,10,14-hexadecapentaenoic acid of the formula 1 or 8101630-14 - a pharmaceutically acceptable salt thereof and a pharmacological carrier. Use of the material according to claim 1 or 5, respectively. the product of the method of claim 2, 3 or 4 for the prevention or treatment of cancer, Use of the material of claim 1 or 5, respectively. the product of the method of claim 2, 3 or 4 for treating keratinization associated skin diseases. 8101630? , £ X - CH2 - CC ^ R, 3 CHO 5 x \ A / CO ^, s ^ k / X ^ VxJvX s * o2Y Z 1 sopy I 8101630