FI60204C - FREQUENCY REQUIREMENT FOR THERAPEUTIC THERAPEUTIC PLEUROMUTILINER - Google Patents

Description

ΓβΙ ANNOUNCEMENT rr \ nf \ A jjgrg W <11> UTLÄGGNINGSSKRIFT 602 0 4 • fevB C (45) n - n'tl iy: 'n! Vtty 10 12 17-! 1

Patent oeddelat (S1) Kv.lk?/lnt-CI.3 C 0? Ώ 295/04 FINLAND —FINLAND (21) Pit * n «lh« k.nw * -Pm.nun.eki »tet 79 ^ 061 (22) Htktmiipilvl - AMeknlnpdig 27 * 12.79 * * (23) AlkupUvt — Giltlgh« t »Dag 26.09.72 (41) Has become slipped - Bllvlt offundig 27.12.79

Patent and Registration Office .... " ,., ., ., ..

_. . . . ^. (44) Nihtlvllulptnon> kuvl. - aD q,

Patent-och registerstyrelsen '' Antokin utltgd oeh uti. * Krift «n pubHcund 31.08.81 (32) (33) (31) ttuoikuus — Begird prloritaf 05.10.71 O5.IO.7I, 25.Ο5.72, 25.Ο5 .72, 25-5.72 Swiss Switzerland (CH) II + I + 50 / 7I * 1M51 / 71 7738/72, 7739/72, 77 ^ 0/72 (71) Sandoz AG, Basel, Switzerland ( CH) (72) Helmut Egger, Vienna, Hellmut Reinshagen, Vienna, Austria-Austria (AT) (71 *) Oy Kolster Ab (5U) Method for the preparation of new therapeutically useful pleuromutilins - For the preparation of new pleuromutilins ) Separated from application 261 + 5/72 (patent 58635) ~

Avdelad frän ansökan 261 + 5/72 (patent 58635)

The present invention relates to a process for the preparation of novel therapeutically useful pleuromutilins of the formula __-CO-CH 2 -S- (CH 2) n - / β 2 -R 2 ch 3 oh wherein is ethyl or vinyl, n is an integer from 2 to 5, and R 2 is 260-204 C 1-4 alkylcarbonyloxy-C 1-4 alkyl or benzoyloxy-C 1-4 alkyl and for the preparation of acid addition salts and quaternary salts of these compounds.

According to the invention, novel pleuro-mutilines of the formula I and their acid addition salts and quaternary salts are obtained in such a way that the compound of the formula Ia is O-CO-CH, -S- (CH 2) V 1 -R 2. o I 2 n v_y 2 \ - <“· CH3 oh where R. j and h have the same meaning as above and is lower hydroxyalkyl, is reacted with a compound of formula

, R4-B III

wherein B is alkoxycarbonyl or chloro or bromoformyl, or the group R 1 -CO-O-CO-, and R 1 is lower alkyl or phenyl, and the resulting pleuromutilins of formula I are, if desired, converted into an acid addition salt or a quaternary salt.

The process can be carried out, for example, by reacting a compound of the formula Ia with an acid derivative of the formula III in a solvent. Preferably, the acid derivative can also be used as a solvent, or a mixture of an acid derivative and an inert solvent, e.g. a chlorohydrocarbon such as dichloromethane, is used. The reaction product can be separated from the reaction mixture by methods known per se and purified, if appropriate.

The compounds of formula I can be converted into their acid addition salts and vice versa.

The corresponding quaternary salts can be obtained from the compounds of the formula I by methods known per se.

• · · I

The lower hydroxyalkyl group R 1 preferably contains 1 to 4, and especially 2 carbon atoms.

3 60204

The number n is preferably 2 or 3.

The compounds of the formula I, their pharmacologically acceptable acid addition salts and their quaternary salts, while having very low toxic, interesting biological, in particular antimicrobial, properties, can therefore be used as medicaments.

They have an inhibitory effect on bacteria, which has been found in in vitro (agar plate) and in vivo (mouse experiments) studies using different bacterial strains. This inhibitory effect has always been observed in the concentration range of about 0.002-5 μg / ml. In particular, the inhibitory effect on mycoplasma has been studied and found to be above a concentration of about 0.008-2.5 μg / ml. As a result, the compounds of the invention can be used as effective bactericidal antibiotics.

As medicaments, the compounds of the formula I and their water-soluble, physiologically acceptable salts can be used alone or in suitable dosage forms together with inorganic or organic pharmacologically inert excipients. They are used, for example, as ingredients in capsules and injection or instillation preparations which contain a sufficient amount of active ingredients to achieve an optimal blood count, i.e. about 10-500 mg per capsule.

In addition, the compounds according to the invention form excellent additives for nutrient mixtures and drinking water.

Starting compounds of formula Ia ’

O-CO-CHo-S- (CHo) - / 'n-rJ

LTT j / 1 y CH3 oh where R 1, n and R 2 are as defined above can be prepared by reacting a compound of formula IV 60204 0-CO-CH 2 -O-SO 2 -R 5 fy <r ^ 1/1 iv y - \ CH3 CH3 oh wherein is as defined above and R1 is alkyl or aryl to react with a compound of formula V / ~~ \ i

H-S- (CH2) n-N2 "* 2 V

where n and denote the same as above.

This process can be carried out, for example, by dissolving an acid addition salt of a compound of formula V, e.g. a hydrohalide, or a free base in a solution of sodium in an anhydrous lower alcohol, e.g. ethanol. To this solution is then added a solution of a compound of formula IV in an inert solvent, e.g. an aliphatic ketone such as ethyl methyl ketone or acetone. The reaction preferably takes place at room temperature up to the boiling point of the reaction mixture, especially at 25 to 55 ° C, and lasts for 2 to 12 hours.

Compounds of formula

ψ / Λ - (CH2) n-SH V

wherein n and R 2 are as defined above, can be obtained by reacting a compound of formula VI with a thiourea and then hydrolyzing the complex formed alkalally.

If the starting material products have not been described, they are known or can be prepared by methods known per se, i.e. analogously to the methods described here or by methods known per se.

60204

Pharmacological tests and their results

Method: Determination of the minimum effective concentration by the dilution method (dilution factor 0,5) Medium: Modified Bakos broth (without phenol red) Culture time: 48 hours at 37 ° C in a tube, after which the sample was transferred to an agar plate and a solution containing the test compound was added. , and growth was monitored for 48 hours (37 ° C).

The compounds tested, the test organisms used and the results obtained are shown in Tables I and II.

Table I. Minimum effective concentration (μg / ml) of the compounds according to the invention

Micro-14-deoxy-14-14-deoxy-14-Tetra organism ££ 2- (4-acetoxy- [[2- (4-acetoxycycline ethyl) piperazino] ethyl) piperathydroethyl mercaptoacetate sino-ethyl chloride oxy-4-mutilin captoacetoxy-mutilin δcc: M. hcmirus 14152 0.0078 0.0039 3.125 M. gallisepticum 19610 0.0156 0.0078 0.78 M. m.p. 1 SH | 0.0128 0.625 6.25 M. Hyorhirus 17981 0.0062 0.156 6.25

Table TI. Minimum effective concentration of reference compound (pg / ml)

Microorganism 14-deoxy-14-p-hydroxy-Tetracycline-phenylmercaptoacetoxy-hydrochloride mutilin

ATCC

M. hominis 14152 3.9 3.12 M. gallisepticum 19610 1.95 3.12 M. sp. I R1 15.625 12.5

The results show that the reference compound (DE-OS 2 036 027, Example 16) has almost the same activity as tetracycline hydrochloride and that the new compounds have a significantly better activity.

6 60204

In the following examples, which illustrate the invention in more detail, all temperatures are shown in degrees Celsius.

Example 1; 14-Deoxy-N- [S- (4-acetoxyethyl) piperazino] ethyl] mercaptoacetoxy} mutilin a) (4-Hydroxyethylpiperazino) ethanethiol 4-Hydroxy 4-hydroxyethylpiperazine is heated together with a benzene solution of ethylene sulfide for 17 hours at a molar ratio of 3: ° C. Allow to cool, separate the polymeric material by filtration, wash with benzene, evaporate the solvent, and distill the product through a Vigreux column. The product obtained has a boiling point of 97.5 ° / 0.1 l torr.

b) 14-Deoxy-14β-2- (4-acetoxyethyl) piperazino] ethyl-mercaptoacetoxy-4-mutilin 1.81 g of (4-hydroxyethylpiperazino) ethanethiol are added under nitrogen to a solution of 0.35 g of sodium in 25 ml of anhydrous ethanol. and then dropwise a solution of 5.35 g of 14-deoxy-14-tosyloxyacetoxymutilin in 15 ml of ethyl methyl ketone. After stirring for 4 hours at room temperature, the solvent is evaporated off under vacuum and the residue is dissolved in 50 ml of ethyl acetate, with partial transesterification. A. The separation of the acetylated product from the unacetylated product is carried out by column chromatography on silica gel using chloroform / methanol (7: 1) as eluent. The faster moving fraction is converted to the hydrochloride with a softening point of 137-140 ° C by means of an ethereal solution of hydrogen chloride. The corresponding bis (hydrogen maleate) has a melting point of 142-144 ° C.

Example 2: 14-deoxy-1H-1H. - (4-Acetoxyethyl) piperazino-7-ethylmercaptoacetoxy) -mutiline a) d4-Deoxy-14- (C2- (4-hydroxyethyl) piperazin-7-ethylmercaptoase) The procedure is as described in Example Ib, but is dissolved by evaporation in vacuo. the residue is taken up in 50 ml of methylene chloride, washed with 5 times the amount of water and dried over magnesium sulphate, 5 ml of an ethereal solution of 5.8% hydrogen chloride are added to the solution, and the hydrochloride crystallizes on addition of the ether.

7 60204 b) 14-Deoxy-14fa - (4-acetoxyethyl) piperazino and ethyl mercaptoacetoxy) mutilin to 4.0 g of 14-deoxy-14β- (7- (4-hydroxyethyl) piperazine] ethylmercaptoacetoxy-4-mutilin 10 ml of acetic anhydride are added under ice-cooling and the mixture is stirred for 5 hours at room temperature, then the mixture is poured into 150 ml of cold water, stirred for 1 hour and extracted three times with ether. The precipitated free base is dissolved in ethyl acetate, and after drying the solution over magnesium sulfate and evaporating the solvent, a pure base remains, and the melting point of bis (hydrogen maleate) is 142-144 ° C.

Example 3: 14-Deoxy-14- ({((4'-propionyloxyethyl) piperazino) ethylmercaptoacetoxy} mutilin 0.50 g of 14-deoxy-14-4-hydroxyethyl) piperazino-7-ethylmercaptoacetoxy}} mutilin is boiled in 5 ml. in dichloromethane with 0.20 g of propionyl chloride for 2 hours under reflux After cooling, the crystalline dihydrochloride is precipitated by adding ethereal hydrogen chloride solution and diluting with anhydrous ether, the salt is filtered off with suction and washed with ether, mp 182-187 °.

Example 4: 14-deoxy-14-Jh. - (4-pivaloyloxyethyl) piperazino-7-ethylmercaptoacetoxy) mutilin 0.50 g of 4-deoxy-N- (4- (4-hydroxyethyl) piperazino-7-ethylmercaptoacetoxy} -mutiline is boiled in 5 ml of dichloromethane to 0.15 g. with pivaloyl chloride for 4 hours at reflux. After cooling, the precipitated is taken up in ethereal hydrogen chloride solution, and diluted with anhydrous ether to precipitate a crystalline dihydrochloride. After suction filtration, it is washed briefly with anhydrous ether and dried in vacuo. The compound is very easily hydrolyzed in the presence of water to give the starting material.

60204

Example 5: m-deoxy-m- [2- (4-benzoyloxyethyl) piperazino] ethylmercaptoacetoxy mutilin 0.50 g of 14-deoxy-14-4-hydroxyethyl) piperazino-ethylmercaptoacetoxy-mutilin is reacted as described in Examples 3 and 4. with 0.16 g of benzoyl chloride for 2 hours. The dihydrochloride is then precipitated as described. Softening point 132-135 °.

Example 6: 14-Deoxy-14-4-acetoxyethyl) piperazino-7-ethylmercaptoacetoxy-4-dihydromutilin a) 14-Deoxy-1H-tosyloxyacetoxydihydromutilin

Method «C:

To a solution of 8.86 g of dihydropleuromutilin in 30 ml of dry pyridine at -15 ° is added 6.10 g of p-toluenesulfochloride in one portion with vigorous stirring, then stirred for 2 hours at -15 ° and then for a further 1 hour at -15 °. 0 °. The mixture is then poured into ice water and the product is dissolved in methylene chloride. The organic phase is washed while cooling first with ice water and then with saturated sodium hydrogen carbonate solution. Drying over sodium sulfate and evaporation affords a uniform product with a softening point of 78-80 ° by thin layer chromatography.

Method: 0.5 3 g of 14-deoxy-14-tosyloxyacetoxymutilin Hydrogenate in 10 ml of ethyl acetate, 0.10 g of 10% palladium on carbon as a catalyst, at atmospheric pressure and at room temperature. The calculated amount of hydrogen has elapsed after about 1 hour. The solution is separated from the catalyst by filtration and evaporated to dryness in vacuo. Softening point 78-80 °.

b) 14-Deoxy-14 (4-hydroxyethyl) piperazino-ethyl-mercaptoacetoxy "3-dihydromutilin 14-deoxy-14-tosyloxyacetoxide hydromutilin is reacted analogously to Example 2a with (4-hydroxy-ethylpiperazino) ethanethiol to give 14-deoxy- It-βϊ - (4-hydroxyethyl) piperazino / ethyl mercaptoacetoxy-4-dihydromutilin with a softening point of the dihydrochloride salt of 135-140 ° C.

9 60204 c) 14-deoxy-14-CC2- (4-acetoxyethyl) piperazine (17-ethyl-mercaptoacetoxy) -dihydromutilin.

0.50 g of 14-deoxy-14-4-hydroxyethyl) piperazino / ethylmercaptoacetoxy-4-dihydromutilin is allowed to stand with 1.5 ml of acetic anhydride for 2 hours at room temperature, then diluted with 10 ml of water and stirred for 1 hour at room temperature. to decompose excess acetic anhydride. The solution is extracted three times with ether. From the evaporated solution, the dihydrochloride is mixed with ethereal hydrogen chloride as described in Example 4. Sp. 133-135 °.

Claims (3)

60204 Claim: A process for the preparation of new therapeutically useful pleuromutilins of the formula O-CO-CH 2 -S- (CH 0) -1 H -R-I 2 2 n \ / 2 CH 3 oh wherein is ethyl or vinyl, n is an integer from 2 to 5, and R 2 is C 1-1 + alkylcarbonyloxy-C 1-4 alkyl or benzoyloxy-C 1-6 alkyl, v and for the preparation of acid addition salts and quaternary salts of these compounds, characterized in that the compound of formula Ia O-CO-CH 0 -S- (CH 2) - N 'N-R 1 2 2 n \ __ / 2 5% LiJC i \ CH3 ch3 oh where Rj and n · are as defined above and R1 is lower hydroxyalkyl is reacted with a compound of formula R4-B III wherein B is alkoxycarbonyl or chloro or bromoformyl, or the group R 1 -CO-O-CO-, and is lower alkyl or phenyl, and the pleuromutilins of the formula I obtained are, if desired, converted into an acid addition salt or a quaternary salt.