SE446006B - (20S) 3ALFA - / (AMINOACETYL) -AMINO / -5ALFA-PREGANAN-20-OL AND ADDITIONAL SALTS THEREOF, AND PHARMACEUTICAL COMPOSITIONS CONTAINING THESE - Google Patents

Description

The invention thus also relates to (208) 3α-β-Ramino-acetyl) -aminyl-Su-pregnan-20-ol and its addition salts with the pharmaceutically acceptable acids as medicaments. The invention relates to pharmaceutical compositions which, as active main ingredient, contains at least one medicinal product as defined above. These compositions can be administered in particular orally, rectally and parenterally. They may be solids or liquids and be in the pharmaceutical forms commonly used in human medicine, such as e.g. tablets, single or coated, capsules, granules, suppositories, injectable preparations. They are produced by the usual procedures. .

The active ingredient or ingredients may be incorporated therein with excipients commonly used in these pharmaceutical compositions, such as talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, vehicles, aqueous or not, fats of animal or vegetable origin, paraffin derivatives, glycols, wetting, dispersing or emulsifying agents, preservatives.

The dose administered varies depending on the condition being treated, the individual being treated, the mode of administration and the intended product. It can e.g. be between 1 and 100 mg / day when administered orally to the adult.

The compounds defined above can be prepared by reacting funtumidine or (20S) 3α-amino-5H-pregnan-20-ol with an amino acid of the formula O HO-C-CH 2 NHR II wherein R represents a protecting group which is readily is digested, in particular by hydrogenolysis, to give a compound of formula III which is reacted with a cleavage agent for the R group to produce the desired compound of formula I OH which, if desired, is reacted with an acid to produce the salt thereof.

As an R-group, one can e.g. use the carbobenzyloxy group or the carboter-butyloxy group. The invention relates in particular to a process which is characterized in that the protecting group R is the carbobenzyloxy group.

The reaction between (20S) 3α-amino-5α-pregnan-20-ol and the amino acid of formula II can take place in the presence of a condensing agent.

The condensing agent naturally has the task of activating the acid function of the amino acid of formula II ä? H0-C-CH 2 NHR II 8005309-3 As the condensing agent one can use a carbodiimide of the formula R -N = C = NR wherein R 1 and R 2, the same or different, represent an alkyl group having 1 to 8 carbon atoms, and optionally are carriers of a dialkylamino group, or R 1 and R 2 represent a cycloalkyl group.

Among the latter, e.g. mention is made of dicyclohexylcarbodiimide or 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide.

It is also possible to use an alkyl chloroformate such as e.g. methyl chloroformate or ethyl chloroformate. It is also possible to use an alkyl pyrophosphite such as e.g. ethyl pyrophosphite. As the condensing agent, 1-ethyl-3- (3-dimethylamino-propyl) -carbonylamines are preferably used.

As the cleaving agent, hydrogen is preferably used in the presence of palladium.

The salts are prepared by the usual procedures, e.g. by the addition of acids. 7 The starting product used, namely (208) 3u-amino- ~ 5a-pregnan-20-ol is a known product (see for this purpose the Merck Index, ninth edition, section 4144).

The following examples describe the invention without limiting it.

Example 1 (208) 3a-Liaminoacetyl) -amino] -Su-pregnan-20-ol (chlorohydrate and base) step A (205) 3u-Zibenzyloxy-carbonylamino-acetyl) -amineQ7-5a- -pregnane-20- ol 5.2 g (205) of 3u-amino-Su-pregnan-20-ol and 5.2 g of N-carbobenzuloxyglycine are dissolved in 150 ml of chloroform and 15 ml of pyridine. The solution is stirred in an ice bath under nitrogen, and 3.9 g of 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride are added. After 1 hour, 310 mg of 1-ethyl-3- (3-methylaminopropyl) -carbodiimide hydrochloride are added again, and the stirring is maintained for half an hour in an ice bath, after which dilute 8005309-3 with a saturated solution of sodium hydrogencarbonate. The insoluble material is filtered, washed separately with a sodium bicarbonate solution, with water, with 1 N hydrochloric acid and with water and then dried at 500 DEG C. 5.1 g of insoluble material are thus obtained. The chloroform phase is separated from the filtrate and then washed with water, 1 N hydrochloric acid, with water, dried and distilled to dryness. 4.6 g of semi-crystallized residue are obtained. The insoluble matter (5.1 g) and the dry extract (4.6 g) are combined and stirred into a dough with 25 ml of methanol and refluxed. It is cooled with ice, centrifuged, washed with methanol and dried at 500 DEG C. Thus 7.05 g of the desired product with a melting point of 250 DEG-252 DEG C. are obtained, which is used as it is in the following steps. . The analytical sample, m.p. 254 DEG C., is obtained by recrystallization from acetic acid.

Step B Hydrochloride of (20S) 3 - (- Llaminoacetyl) -aminq] -5d.- pregnan-20-ol. 7 g of the product obtained in step A are dissolved in heat in 150 ml of acetic acid. Stir the resulting solution under nitrogen bubbling, cool to 45 ° C and add 700 mg of palladium on 10% carbon. The nitrogen bubbling is replaced by a light hydrogen bubbling. After 1 hour, a stream of nitrogen is replaced by a stream of hydrogen, and the bubbling is continued for 15 minutes. The catalyst is then filtered and rinsed with acetic acid. The filtrate is evaporated to dryness. The crystallized product is dissolved in 50 ml of methanol. By adding 4 ml of a 5.5 N hydrochloric acid solution in ethanol, the desired precipitate precipitates.

The suspension is diluted with 50 ml of absolute ethanol. It is concentrated in half and allowed to stand overnight at +50 C. It is centrifuged, washed with ethanol and dried. 4.43 g of the desired product are obtained.

The filtrate, added with 0.5 ml of a 5.5 N hydrochloric acid solution in ethanol and concentrated to a small volume gives a second yield of 0.35 g of the desired product. The desired product is obtained in pure form with a melting point of 2710 DEG C. after recrystallization from 80% ethanol. 8005509-3 / az / âo = + 19 ° s 1 1 ° (1%, pyrene with io% water) Step C Preparation of the base.

The crude hydrochloride obtained in step B has been used. The crude hydrochloride is dissolved in hot water. By adding an excess of 2 N sodium hydroxide, the base precipitates. It is centrifuged, washed with water, dried and recrystallized from methanol.

Thus, the desired pure product is obtained with a melting point of 269 ° C. chlorohydrate, hereinafter referred to as product A, performed in comparison with Levamisole.

-Levamisole is a known product (Merch Index ninth edition, section 8949), which has immunoregulatory properties, Cancer Research 35 927 (1975) or New England Journal of'Meaicine 289 (21) 1148 (1973).

A Strengthening of IgE production. 7 Female rats weighing 28-30 g are immunized subcutaneously with ovalbumin, mixed with alumina on days 0 and 14, and their serum is taken on day 21 to determine the IgE antibodies formed. The respective injected amounts of the antigen (ovalbumin) and the adjuvant (alumina) are selected so that the production of antibodies is minimal.

The tested compounds are administered subcutaneously 3 hours before the first immunization.

The determination of IgE is performed with passive cutaneous anaphylaxis test. This test consists in causing, by intravenous administration of the antigen in an animal, an antigen-antibody reaction to a skin site where antibodies produced in another animal with the same antigen have been previously injected. This reaction is illustrated by a dye injected simultaneously with the antigen. Suddenly, a colored spot appears at the injection site for the antibodies, evidence 8005309-3 of a sensitized cell and increased capillary permeability as a result. The greatest dilution of the serum is determined, which gives a spot with a diameter of ll - 13 mm in all animals. The appropriate dilution of the serum is injected intradermally in a volume of 0.1 ml to male rats with a mean weight of 250 g in the dorsal region. 48 hours later, the animals receive intravenously 0.5 ml of a solution containing 0.5% ovalbumin and 1% Evans blue in isotonic sodium chloride solution. 30 minutes after this injection, they are killed by bleeding, and the diameter of the blue spot is measured on the inside out.

The following results were obtained: product A 0.5 mg / kg for subcutaneous administration Levamisole 20 mg / kg for subcutaneous administration Conclusion: in this test, product A is much more immunostimulatory than Levamisol. _ B Chronic arthritis caused by adjuvant.

Injection of Freund's adjuvant (Mycobacterium butyricum 6 mg / ml in Bayol 55) into the hind paw causes primary inflammatory damage in rats, then after a latency period of 13 - 15 days initiation of secondary inflammations in the uninjected hind paw and in the front paws, tail and ears .

This secondary arthritis can be compared to human rheumatoid arthritis, as autoimmune reactions are thought to be associated with its determinants.

Male rats aged 42-50 days receive an injection in the sole of the foot of 0.10 ml of Freund's adjuvant.

The treatment begins on the day of the injection of the adjuvant and lasts until the animals are killed, which takes place on the seventeenth day.

The criteria for evaluating the activity of the substances are generally: - the weight gain of the animals, always slowed down in proportion to the intensity of the arthritis; - the volume increases of the injected and non-injected hind paws in relation to the average volume of the corresponding 8005309-3 paws of the control animals; - the arthritis of the front paws, the excessive volume of which is not suitable for a plethysmometric measurement and is thus determined subjectively from 0 to 3 depending on the intensity of inflammation; _ - arthritis of the ears and tail, rated l or 0 depending on the presence or absence of nodules.

In the present study, the effect on secondary inflammations was always observed, i.e. on arthritis in the uninjected hind paw, the arthritis in the hind paws and the inflammations in the ears and tail. w. The active dose, which reduces the secondary inflammations by at least 50%, was determined.

The results were as follows: product A active dose 1 mg / kg (subcutaneous administration) Levamisole active dose 50 mg / kg (oral administration) Conclusion: product A is thus very active against secondary phenomena of arthritis, caused by adjuvant, while a very high subtoxic dose of Levamisole is required for a comparable effect to be observed.

Example 3 Examples of pharmaceutical compositions.

Tablets have been prepared with the following composition: V 'product according to Example 1 (chlorohydrate, 10 mg excipient' (talc, starch, magnesium stearate) q.s.p. 1 tablet

Claims (2)

8005309-3 a I Parehtkrav (208) 3u-N- (aminoacetyl) -amino; 7-Sa-pregnan-20-ol and addition salts thereof with the organic acids or mineral acids. Pharmaceutical compositions having an immunostimulatory effect, characterized in that they contain as active ingredient (ZOS) 3u-Å_ (aminoacetyl) -amino_7 ~ 5a- -pregnan-20-ol or addition salts thereof with the pharmaceutically acceptable acids such as drugs.