SU1327789A3 - Method of producing steroid derivatives with substituted 3-aminogroup or acid additive salts thereof - Google Patents

Abstract

The title compounds of formula I and their salts are, with certain exceptions, new: <IMAGE> W is H or OH; X is ethyl, 1-hydroxyethyl, or acetyl; or W + X = ethylidene; R, is H or Me; R2 and R3 are each H, alkyl or hydroxyalkyl, R3 alternatively being acyl (including alkoxycarbonyl and radicals derived from amino acids and peptides), and at least one of R2 and R3 being other than H. Preparation: from the corresponding primary amines (R2 = R3 = H) Use: treatment of autoimmune diseases. <IMAGE>

Description

one

This invention relates to a process for the preparation of novel steroid compounds, specifically to steroid derivatives with a substituted 3-amino group of the general formula

SNS X (I) Rlf -f - W

% KG

where w

: N

Xai

hydrogen or hydroxyl group;

- group CH-x or CH

one

X

sn-he

R is hydrogen or methyl;

R is hydrogen or methyl;

RJ is a hydrogen or oi-amino-carboxylic acid radical, an alanine residue, glutamic acid, serine, phenyl alanine, tryptophan, proline, or a peptide residue containing 2-3ci---mined acids, glycyl-glycine, with this, if at the same time R is methyl, and W and Rg are hydrogen, and X is the CH radical

in this case, Rj is not a mo

It can be a residue of a CS-aminocarboxylic acid, if at the same time R is methyl, RJ and W are hydrogen, and X is a group

HE

the hydroxyl of which has the configuration (S), and the amino group is in the ZL- position, then R is not mo. It can be a glycyl-glycine radical, or their acid addition salts with immunotherapeutic properties.

The purpose of the invention is to obtain new derivatives of steroids that have pharmacological advantages over known structural analogues.

The invention is illustrated by the following examples.

Example 1. (2S) 2-amino-L- - (20S) -20-hydroxy-5 |) 6-progenan-III-ylJ propanamide (hydrochloride and base).

Stage A: 2- (1,1-dimethylethoxycarbonyamino) -N- (205) -20-hydroxy- (5l) pregnan-3 (-yl propane amide.

1.92 g (20S) of Zo-amino-3c-Pregnan-20-ol and 2.27 g of tert-butyloxy carbonyl-b-alanine are dissolved in 60 cm of chloroform and 12 cm of pyridine. Pere- -

a 327789. 2

stir the solution in an inert atmosphere at 0-5 ° C and add 1.1 A g of 1-ethyl-3- (3-dimethylamide-nopropyl) -carbodiimide hydrochloride. After 1 hour and 15 minutes, 1.15 g of hydrochloride-1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide is added anew, stirred for 50 minutes at 0-5 ° C, dried, taken in water sodium bicarbonate solution, extracted with ethyl acetate, washed with saturated aqueous sodium chloride solution and then with normal aqueous solution of hydrochloric acid and then again with saturated aqueous solution of sodium chloride, dried, crystallized in isopropyl ether, sucked off, washed with isopropyl ether, dried

20 at a reduced pressure, and get the target product, so pl. 171 ° C. .

Stage B: hydrochloride and base (28) 2-amino-N- (208) -20-hydroxy-25 - (5oi) -pregnan-3 (xi-shl-propanamide.

Option 1. In an inert atmosphere, 1.55 g of the product obtained at station A in -100 cm of nitromethane are introduced into a suspension, hydrochloric acid is bubbled in for 10 minutes, stirred at 20–25 ° C for 35 minutes, the excess of hydrochloric acid is removed, sucked off, washed with ethyl ether, dried at 60 ° C under reduced pressure of 35 mercury, recrystallized in methanol, and 990 mg of the desired hydrochloride are obtained, m.p. 220 ° C.

Option 2. In an inert am; In the first sphere, 2.7 g of the product obtained in days 40 A are dissolved in 10 cm of ethanol, 40 cm of hydrochloric ethanol (3.5 N) is added, stirred for 8 hours, dried, taken up in ethyl acetate, and filtered off with suction. - washed, washed with ethyl acetate, dried at 60 ° C under reduced pressure, recrystallized in methanol, and 2.2 g of the expected hydrochloride are obtained, mp. 220 ° C.

Preparation of the base. 1.74 g

hydrochloride is dissolved in 100 cm of tetrahydrofuran containing 30% water, 4 cm 2 n is added. caustic soda, brought to dryness, taken up in 100 cm of chloroform, washed with water, dried, recrystallized in ethyl acetate, and obtain 1.27 g of the desired base, m.p. 224 C.

According to the proposed method

: l

BOCK-b-Glutamino | (f CH-CHi-CH COOH ISI

PL ST ST If MG YEARS

wa acid

NHi CH-CH20H 1SI

(chlorine

WOCK-b-serine

The derivatives described in Table 1 were obtained.

Table 1

isi he

: l

 | (

 | (f CH-CHi-CH COOH ISI

NHi CH-CH20H 1SI

. IN

(hydrochloride)

210 (hydrochloride)

BOC-L-Phenyl Alanine

Side-by-trypto fan

240

FSF (hydrochloride)

ISI

135 (base)

WOCK-l-Proline

CFj COOH

BOK-glycyl-glycine

The second function of the acid, L-glutamic acid, is blocked as a benzyl ester; it is released by catalytic hydrogenation in acetic acid in the presence of palladium before elimination of BOC (tert-butyloxycarbonyl).

Example 8. 2-Amino-N- (20S) -17 hydrochloride (i, 20-dihydroxy-5 1 -preg-nan-Css-yl-acetic amide.

Analogously to example 1 (option 2), starting from BOC-glycine and (20S) CcC-am-; but-5 (-pregnan-1 7 (1, 2b-diol, get the target product, t.pcs a; 200 C. (D6) v ° (to 1.5%, ethanol 95),

/ SI

275 (hydrochloride)

Sn, 2-N-CO-SNg-IN2

240

(hlorgidI at) 200 then

252 (base)

The starting (20S) 3 6-amino-5b6-pregnan-17 ((, a 20-diol can be obtained as follows.

Stage 1: (d) - (5U-Pregn-17- (20) - -En-3 -ol.

To 16.1 g of potassium t-butylate in a solution of 160 cm of tetrahydrofuran, 59.4 g of bromide triphenyl ethylphosphonium were added, stirred for 30 minutes, 23.2 g of epidrosterone were added, stirred for 15 hours, poured into ice water, extracted with ethyl acetate, washed with water, dried, distilled to dryness, purified by chromatography 513277896

Cesky on silicon dioxide (elant - tylen and 16.5 cm of pyridine cool

before, enter

cyclohexane - ethyl acetate (7: 3), crystallized in methanol, ice nt, sucked off, dried, and 23.1 g of the expected product are obtained,

m.p. .

Stage 2. (g) -Zy4azido (5 ()) - Pregn-17-17 (-20),

To a solution of 1.66 g of this product for 5 minutes, 16.5 cm of trifluoroacetic anhydride, stirred for 15 minutes at room temperature, distilled to dryness under reduced pressure, added 200 cm of water, sucked off, washed with water, dried under reduced pressure of 18, 1 g of crystals.

nie and get - About so pl. 204 ° C.

In 30 cm of benzene and 5 cm of rofuran tetrahydrate, 1.92 g of ethyl azodicarboxylic ester and 3.02 g of azidophosphoric acid diphenyl ester are added, the mixture is stirred in an ice bath, a solution of 2.88 g of triphenylphosphine in 30 cm of benzene is added stirred for another 40 minutes at 10 ° C5 washed with a solution of 2 n. hydrochloric acid and then with water, dried and

-

-

- 2

the mixture is dried to dryness, purified by chromatography on silica (eluent is heptane, and then heptane is ethyl efficiency 1: 1) and 1.67 g of crystals of the target product are obtained, mp. 114 ° C after recrystallization in methanol.

Stage 3: (d) -5o (, - Pregn-17 (20) -en-Zo-amine (and hydrochloride).

In 290 cm of tetrahydrofuran is dissolved 14.5 g of the obtained above (Z) - -Zo (, - azido- (5a,) - Pregn-1 7 (20) -en (in this form), mixed by heating at 25-27 0 800 mg of lithium aluminum hydride is added over 1 hour, stirred for another 1 hour, excess hydride is removed with methanol, filtered, the filtrate is washed with an aqueous solution of ferro salt, and then saturated with sodium chloride, dried, distilled off and dry. 13.1 g of crystals of the desired amine are obtained, mp.

Dissolve the base in 150 cm of ethyl acetate and 90 cm of methylene chloride, add 27 cm of 1.7 n. acetic acid ethyl ester, sucked off, washed with acetic acid ethyl ester and obtained to: the crystals dried under reduced pressure to obtain 13.2 g of the crystals of the hydrochloride of the target product, so pl. 300 C.

(ii) j at 1% in pyridine, at 10% in water +38.5 ± 1.5.

Stage 4. N- (Z, 5jC) -nperH-17 (20) -en-Zsb-yl-trifluoroacetamide.

In an inert atmosphere, 16.5 g of the hydrochloride obtained in stage 3 in 165 cm of honey are introduced into the suspension, injected

for 5 minutes 16.5 cm of trifluoroacetic anhydride, stirred for 15 minutes at room temperature, distilled to dryness under reduced pressure, add 200 cm of water, suction, rinse with water, and dry under reduced pressure to 18.1 g of crystals.

g and get Oh so pl. 204 ° C.

15

20

-

- WITH

25

thirty

Stage 5: N- (20S) -17 ", 20-dihydroxy- (5 ° C) -pregnan-Zy-yl-trifluoroacetamide.

In an inert atmosphere, 18.1 g of the product obtained are dissolved in 100 cm of methyl ethyl acetone, 9 g of trimethyl-amine N-oxide dihydrate are added, and also, as a solution of 360 mg of four osmium fetoxide in 71 cm of methyl acetone, stirred for 2 hours - flux, allowed to cool, add 200 cm of 10% sodium thiosulfate solution in water, stir for 30 minutes at room temperature, decant, wash with water, dry over magnesium sulfate, filter, and distilled to dryness under reduced. pressure, purify the resulting oil, chromatographically on silica (eluant-benzene-ethyl ester of acetic acid 7: 3) and obtain 14 g of the desired product, so pl. 172 ° C and then

192 C.

35

4Q

45

 6. (205) -3 (-amino-5 (), - pregan-17 (, 20-diol.

In an inert atmosphere, 4 g of the product obtained above is dissolved in 20 cm of methanol, 8 cm of sodium hydroxide is added, stirred for 1 h 30 min, 50 cm of water are added, stirred for 10 min, sucked off, washed with water, dried at 40 under reduced pressure and get 3 g of the target product, so pl. 210 C.

Example 9. 2-Amino-H- (205) -20-hydroxy-19-nor- (5-6) - -pregnan-3 (-yl-acetamide) hydrochloride

Analogously to example 1 (option 1), Q but starting from BOC-glycine and (20S) -amino-19-nor- (5 () -pregnan-20-ol, the desired product is obtained, mp = 270 ° C by sublimation, (ot) "39.5 ° i: 1.5 ° (to 1% methanol). Source (203) -3-o-amino-19-nor- - (5o (,) - pregnan-20-ol can be obtained as follows.

Analogously to example 8 (steps 1-3) to obtain (Z) -5-ti-nperH-1 7 (20) 713277898

-eH-3ei-aMHHa, isocyan from 19-nor-epian. Example 11. 2-amidrosterone hydrochloride was obtained (Z) -5 (i-19-Hop-. (20R) -20-hydroxy-5 (x -pregnannegreg -1 7 (20) -en-3b-amine.

156 mg of sodium borohydride in 5 cm of tetrahydrofuran were introduced into the suspension under nitrogen atmosphere, a solution of 0.5 cm of ethereal extraction of boron trifluoride in 2.5 cm of tetrahydrofuran was added dropwise at 5 ° C, stirred for 1 hour in an ice bath, and added 296 mg of (Z) -5 () i-19-Hop-nperH-17 (20) -en-3-amine-amine in a solution in 3 cm of tetrahydrofuran are mixed, stirred for 1 h 30 min at room temperature, cooled on ice 2 cm 6 and are added slowly. of sodium hydroxide solution, stirred for 5 minutes at room temperature, decanted, the aqueous layer was extracted with tetrahydrofuran, the organic layer was washed with water, 4 cm 5 n were added. caustic soda, 2 cm 110 volume hydrogen peroxide, stirred for 45 minutes, extracted

-Zo Il J-acetamide,

Analogously to example 1 (option 1), 5 but starting from BOC-glycine and (20R) -3ei- -amino-5 (1b-pregnan-20-ol, the desired product is obtained, mp 210 C (then 260 C) . (, + 22 ° t G (to 0.8%. Pyridine containing 10% water), fO Example 12. Chlorohydrate 2 (S) 2-, -amino-M-methyl-K- (20S) 20-hydroxy - -5 (-pregnan-3 () 6-yl-propanamide.

Expose (20S) -3ti -methylamino-5b6 pregnan-20-ol to BOC-5 -L-alanine of Example 1 (option 1) to obtain the desired product, the hydrochloride of which is also prepared, so pl. 270 ° C.

Example 13. 2 (R) 2-20-amino-N-methyl-N- (205) -20-hydroxy-5 -6-6-pregnan-3O6-yl-propanamide hydrochloride.

(20S) -Th-methylamino-5a {-pregnan--20-ol is reacted with BOC-b-alanine of Example 1 (option

ethyl acetate, pro-25 1) to obtain the target product, washed with water, dried and distilled to dochloride, which is prepared dry under reduced pressure. A dry extract is taken in 10 cm meta (sublimation at about 260 s).

Example 14. Obtaining (2S) -2NOL with 5 cm 1 n. hydrochloric acid.

Example 14. Obtaining (2S

Heat 30 minutes in a water bath at 30-amino-N (205) -20-hydroxy-5y-pregnen50 50, drink into a saturated solution (1-H-indole-3-propanamide).

sodium bicarbonate, extracted with methylene chloride, washed with water,

dried, scaled under reduced

pressure and get 257 mg of crystal

fishing target product, so pl. 190 ° C.

According to Example 1 (Option 2), starting from 3-amino-5-6-pregnan-20-ol and tert-butoxycarbonyl - (.- alan

, get the target product, tp 140 ° C and ((i) j -34 1 ° (1% CHClj)

Example 15. i-Dimethylamino -N- (20S) 20-hydroxy-5 (-pregnane-3-yl-acetamide.

Example 10. 2-Ami-Ho-N- (1.7 ° (. - hydroxy-20-oxo-5c6-pre-benan-3 (1 / -yl) acetamide) hydrochloride.

Analogously to example 1 (option 2), starting from BOC-glycine and So6-amino-176-hydroxy-5 (-pregnan-20-one, the desired product is obtained, mp. "

(about) + 50 ± 1 ° (to 1% ethanol

95). ..

I

The starting 3 {-amino-17ei-hydroxy-5 () 6-pregnan-20-one can be obtained by the following method. .

The nadhromic oxidation of the product obtained in Example 9 in stage 5 gives N- (1 7 "; - hydroxy-20-oxo-5

The introduction of the animal product, prompting the activity of the immune sispregnan-3 / -yl) -trifluoroacetamide,

m.p. 178 C (then 18b C), which 06–55 is expressed by an increase in shock

The test is performed in accordance with Example 9 in Stage 6, the response to the introduction of the intigen, to the cathode of preparing the So6-amino-17th hydroxy-rum, was sensitized. -5 () C-pregnan-20-she, so pl. (male mice male weighing 30-35 g of water crystallization sensibil).

-Zo Il J-acetamide,

Analogously to example 1 (option 1), but starting from BOC-glycine and (20R) -3ei- -amino-5 (1b-pregnan-20-ol, the expected product is obtained, mp 210 C (then 260 C)). (, + 22 ° t G (to 0.8% pyridine containing 10% water), Example 12. Hydrochloride 2 (S) 2- ,.-amino-M-methyl-K- (20S) 20-hydroxy- - 5 (-pregnan-3 () 6-yl-propanamide.

Expose (20S) -3ti -methylamino-5b6 pregnan-20-ol to BOC-L-alanine of Example 1 (option 1) to obtain the desired product, the hydrochloride of which is also prepared, mp. 270 ° C.

Example 13. 2 (R) 2- -Amino-N-methyl-N- (205) -20-hydroxy-5B6-pregnan-36-yl-propanamide hydrochloride.

(20S) -Th-methylamino-5a {-pregnan--20-ol is reacted with BOC-b-alanine of Example 1 (option

1) to obtain the desired product, the hydrochloride of which is prepared

1) to obtain a target product whose hydrochloride prepared

(sublimation at about 260 s).

to obtain the desired product, the hydrate of which is prepared

Example 14. Obtaining (2S) -2-3 -yl- (1-H-indole-Z-propanamide).

According to Example 1 (Option 2), but starting from 3-amino-5-6-pregnan-20-ol and tert-butoxycarbonyl - (.- alany, the desired product is obtained, mp 140 ° C and ((i) j -34 1 ° (1% CHClj).

Example 15. i -Dimethylamino -N- (20S) 20-hydroxy-5 (-pregnane-Zo, -yl-acetamide.

The process is carried out analogously to example 1 (steps A), using 2.522 g (20S) (, α-amino-5 (6-pregnan-20-ol and 3.350 g of N, N-dimethyl glycine hydrochloride. 4 g of crude product are obtained Compound and I, N-disubstituted derivative. A product obtained by spraying with caustic soda in methanol gives the desired product, mp ((Oj +29 t 1 (С 1% CHjClj) ..

Pharmacological study. 1. Auxiliary means of anaphylactic shock.

The administration to animals of a product that induces the activity of the immune systems is expressed by an increase in shock

using serum albumin bull. After 8 days, an antigen is obtained by intravenous route. In terms of Minimal sensitization, control animals do not show a deadly shock during the last administration. The test product is injected intradermally in admixture with antigen: if this product. is an auxiliary agent, fO then it increases sensitization and then a mortal shock follows after administration by the intravenous route.

As the active dose, a dose is taken that causes mortality equal to or higher than 50% of the animals. The results are shown in Table. 2

table 2

2, Test on sockets formed by red blood bodies of a ram. The introduction of a product that stimulates the activity of immunity systems to animals increases their ability to inject the product of immunization.

Male rats aged 3 months. they are sensitized by the intraperitoneal route using b-aran erythrocytes (; a, day 0), 7 days later (day 7) their spleen is withdrawn and splenocytes are brought into contact with sheep erythrocytes, then the percentage of leukocytes around which erythrocytes form rosettes are counted.

The test product is administered by mouth daily.

An immunostimulating dose is that dose of the product, which multiplies by approximately 2 percent of the sockets observed in control animals. The results are shown in Table. 3

Table

Example

Dose per animal in mg / kg

5 6

9 10

Through the mouth 1

2 5

5 5

3. Acute toxicity study. Lethal doses. DL of the various compounds studied were evaluated after oral administration to mice. The dose of DLp is the maximum dose that does not cause mortality after 8 days. The results are given in table 4.

II, Table A

p1327789

Continuation of table 4

ten

1000

eleven

: .400

The dosage, which varies depending on the derivative used, can be from 10 mg and 1 g per day when applied by mouth (men).

Isoprinosine, taken as an analog, showed the absence of a test at a dose of 5 mg / μg. In test 2 for this product, an immunostimulating dose exceeding 50 mg / μm is noted.

Thus, the target compounds are more active than isoprinosine, which has similar biological activity.

Claims (1)

Formula and zo bren The method of producing steroid derivatives with a substituted 3-amino group. sing of general formula (I) --W de W is hydrogen or hydroxyl group; CH3 or pvr group X CH R is hydrogen or methyl; R is hydrogen or methyl; HE 89 R. 12 hydrogen or radical a: -aminocarboxylic acid, the residue of alanine, glutamic acid, serine, phenylalanine, tryptophan, proline or the residue of a peptide containing 2-behind-aminated acids, glycyl-glycine, at that, if R is methyl, W and Rg - wild hydrogen, and X - rasNz in this case R, can not be a residue of cL-aminocarboxylic acid, if at the same time R is methyl, RJ and W are hydrogen, and X is a group - CH .. hydro CH-OH which has the S configuration and the amino group is in the 3tl- position, then RJ cannot be a glycyl-glycine radical, or their acid-additive salts, such that the amine of the general formula (II) ShgH   -w rfW H21ML4J where W, X and R are as defined, are reacted with (L-amino acid or peptide containing 2- or 3-amino acids, whose amino functions are protected by tert-butoxycarbonyl, then the protecting group is removed by acid hydrolysis to obtain the product of general formula (I) and the target the product is added and, if desired, converted to salt. Editor M.Nedoluzhenko Compiled by I. Fedoseeva Tehred L. Oliynyk Proofreader V. But ha Order 3398/58 Circulation 347Subscription VNIIPI USSR State Committee for inventions and discoveries 113035, Moscow, Zh-35, Raushsk nab., 4/5 Production and printing company, Uzhgorod, st. Project, 4