CH633300A5 - Spiramycin esters - Google Patents

Abstract

The spiramycin esters correspond to the formula below, in which R<1> = H, acetyl or propionyl, A = butenylene or butadienylene with trans double bonds, and R<2> = COZCOYR<3> (Z = alkylene, Y = S or O, R<3> = alkyl, cycloalkyl, aryl or aralkyl), and to their acid addition salts. Applications as antibiotics against Staphylococcus aureus, pneumococci, streptococci, in veterinary medicine for the treatment of mycoplasmosis, and as growth stimulating factors for animals. <IMAGE>

Description

       

  
 

** ATTENTION ** start of the DESC field may contain end of CLMS **.

 represents an alkyl group containing from 1 to 4 carbon atoms and from an acid acceptor, at a temperature of - 30 to + 30 C and in the presence of an anhydrous inert organic solvent, and, where appropriate, it is transformed the compound obtained in one of its salts by reaction with an acid.



   4. Process for the preparation of the compounds of the general formula (I) according to claim 1 and in which the symbols Rt, A and R2 are as defined in claim 1 and their acid addition salts, characterized in that that compounds of general formula are reacted:
EMI2.1
 in which R1 and A are as defined in claim 1, with compounds of general formula:
 X2OCZCOYR3 (VIII) in which X2 represents a bromine or chlorine atom or a group of general formula:

  :
 -OCOOR5 (IX) in which R5 is as defined in claim 3 and Z, Y and
R3 are as defined in claim 1, in the presence of an acid acceptor or of a mineral base, at a temperature of -30 to +30 "C and in the presence of an anhydrous inert organic solvent, and, where appropriate, the compound obtained is converted into one of its salts by reaction with an acid.



   5. Process for the preparation of the compounds of general formula (I), in which the symbol A represents a trans butenylene group and the symbols R 'and R2 are as defined in claim 1, characterized in that a compound of general formula (I) in the formula of which the symbol A represents a butadienylene group, in which the double bonds are trans, and the symbols R1 and R2 are as defined in claim 1, by the method according to claim 3, and that it is partially reduced by catalytic hydrogenation.



   6. Method according to claim 5, characterized in that a noble metal or one of its oxides is used as catalyst, either alone or deposited on alumina, charcoal or barium sulphate.



   7. Process for the preparation of the compounds of general formula I in which the symbol A represents a trans butenylene group and the symbols Rl and R2 are as defined in claim 1, characterized in that a compound of general formula is prepared (I) in which the symbol A represents a butadienylene group, in which the double bonds are trans, and the symbols R1 and R2 are as defined in claim 1, by the method according to claim 4, and that it is reduced partially by catalytic hydrogenation.



   8. Method according to claim 7, characterized in that a noble metal or one of its oxides is used as catalyst, either alone or deposited on alumina, charcoal or barium sulphate.



   9. Pharmaceutical composition comprising, as active ingredient, at least one spiramycin derivative according to claim 1, or one of its non-toxic acid addition salts, in association with a compatible and pharmaceutically compatible support or coating. acceptable.



   10. Composition according to claim 9, characterized in that it comprises ethylsuccinate-2 'of a mixture of spiramycins containing approximately, by weight, 60% of spiramycin I, 20% of spiramicin II and 20% of spiramicin III .



   The present invention relates to novel antibiotics, methods of preparing them and pharmaceutical compositions containing them.



   The present invention relates to new spiramycin derivatives of general formula:
   (Formula at the head of the next page) in which R1 represents an acetyl or propionyl group or, preferably, a hydrogen atom, A represents a butene group or, preferably, butadienylene group (all the double bonds present in the group A being trans), and R2 represents a group of general formula:

  :
EMI2.2
 in which Z represents a bivalent alkylene group, saturated or unsaturated, straight or branched chain, containing from 1 to 6 carbon atoms (for example an ethylene group), Y represents a sulfur atom or, preferably, a d atom oxygen, and R3 represents a straight or branched chain alkyl group containing from 1 to 6 carbon atoms, preferably from 1 to 3, a cycloalkyl group containing from 3 to 8 carbon atoms, an aryl group (for example phenyl) or an aralkyl group (for example phenylalkyl) in which the alkylene residue is straight or branched chain and contains from 1 to 6 carbon atoms, and their mixtures,

   especially mixtures in which the compounds of general formula (I) contained therein differ only with regard to the symbol R1 and are identical with regard to the symbols A and R2, as well as the non-toxic addition salts acid of these compounds.



   The spiramycin derivatives of general formula (I) in the formula of which R2 represents a group of general formula (II) in which Z represents an ethylene or trimethylene group, Y
EMI3.1




  represents an oxygen atom and R3 represents a straight chain alkyl group containing from 1 to 3 carbon atoms, preferably the ethyl group, are the preferred derivatives.



   By the term non-toxic acid addition salt is meant a salt whose anion is relatively harmless to the animal organism when it is used in therapeutic doses, so that the beneficial properties of the cation are not adversely affected by side effects attributable to this anion.



   In the present text, whenever reference is made to the compounds of general formula (I), it should be understood that these are also the abovementioned salts, when the context permits.



   As can be seen, the group of general formula (II) can have a chirality resulting from the chirality of one or both residues
Z and R3. The presence of chirality, as we know, can lead to isomerism. The present invention encompasses in its field all the isomers of general formula (I) originating from said chirality in the groups of general formula (II), as well as their mixtures.



   According to a characteristic of the present invention, the compounds of general formula (I) in which the symbols R ', A and R2 are as defined above are prepared from compounds of general formula:
EMI3.2
 in which R 'and A are as defined above, and R4 represents a group of general formula:
EMI3.3
 in which Z is as defined above, by application or adaptation of known methods for the preparation of esters from carboxylic acids, for example by reaction with compounds of general formula:
 R3YH (V) in which Y and R3 are as defined above.



   The reaction is carried out in the presence of a compound of general formula:
   XICOORs (VI) in which Xl represents a bromine or chlorine atom and R5 represents an alkyl group containing from 1 to 4 carbon atoms, and an acid acceptor, for example a tertiary amine, such as triethylamine, preferably between - 30 and + 30 C, and preferably in the presence of an anhydrous inert organic solvent, for example chloroform.

 

   According to another characteristic of the present invention, the compounds of general formula (I) in which the symbols R ', A and
R2 are as defined above are prepared from compounds of general formula:
 (Formula at the top of the next page) in which R 'and A are as defined above, by application or adaptation of known methods for the preparation of esters from alcohols, for example by reaction with compounds of general formula :
 X20CZCOYR3 (VIII) in which X2 represents a bromine or chlorine atom or a group of general formula:
 -OCOOR5 (IX)
EMI4.1

 in which R5 is as defined above, and Z, Y and R3 are
 as defined above.



   The reaction is carried out optionally in the presence of an acid acceptor, for example a tertiary amine, such as triethylamine, or an inorganic base, such as sodium bicarbonate, preferably in the presence of an anhydrous inert organic solvent, for example chloroform or acetone and, preferably, at a temperature of -30 to +30 C.



   According to another characteristic of the present invention, the compounds of general formula (I) in which the symbol A represents a trans butenylene group, and the symbols R1 and R2 are as defined above, are prepared by partial reduction of the compounds of general formula (I) in which the symbol A represents a butadienylene group (in which the double bonds are trans) and the symbols R 'and R2 are as defined above, by application or adaptation of known methods, for example by catalytic hydrogenation.



   Among the suitable hydrogenation catalysts, there may be mentioned noble metals such as palladium and platinum, and their oxides, optionally deposited, for example, on alumina, charcoal or barium sulfate.



   The non-toxic acid addition salts of the compounds of general formula (I) can be prepared by application or adaptation of known methods for the preparation of salts of organic bases, for example by reaction of the compounds of general formula (I) with the appropriate acid, in a suitable solvent. These salts can be isolated by lyophilization of the solution or, if they are sufficiently insoluble in the reaction medium, by filtration, after removal of part of the solvent if necessary.



   Mixtures of compounds of general formula (I) can be separated into their components by application or adaptation of known methods for the separation of mixtures of antibiotics, for example by a countercurrent distribution method, for example in a Craig (A. Weissberger, Technique of Organic
Chemistry, Interscience Publishers, New York, vol. III, p. 286), by chromatography (for example on alumina or on silica gel), or by fractional crystallization, for example from an aromatic hydrocarbon as solvent, for example benzene.



   The compounds of general formula (III) can be prepared from the compounds of general formula (VII) by application or adaptation of known methods for the preparation of hemicers of dicarboxylic acids, for example by reaction with compounds of general formula:
EMI4.2
 in which Z is as defined above.



   The reaction can be carried out in the presence of an inert organic solvent, preferably at room temperature and optionally in the presence of a base, for example an alkali metal carbonate or bicarbonate, a tertiary amine or a heterocyclic tertiary base such as pyridine.



   The compounds of general formula (VIII) can be prepared from the compounds of general formula (V) by application or adaptation of known methods for the preparation of esters from alcohols and thiols, for example by reaction of the compounds of general formula (V) with compounds of general formula (X), in a manner similar to that described above for the preparation of the compounds of general formula (III) from the compounds of general formula (VII), then transformation hemiesters thus obtained in compounds of general formula (VIII) by application or adaptation of known methods for the preparation of acyl halides or mixed anhydrides from carboxylic acids.



   The compounds of general formula (VII) in which the symbol
A represents a butadienylene group (in which the double bonds are both trans) and the symbol R1 represents a hydrogen atom, an acetyl group or a propionyl group, which are known, respectively, under the names of spiramycin I, spiramycin It and spiramycin III, as well as their mixtures, can be prepared by application or adaptation of known methods for the preparation of spiramycins, for example by aerobic culture of Streptomyces ambofaciens NRRL 2420 or of one of its mutants producing spiramycin.



   Spiramycin usually obtained by fermentation of
Streptomyces ambofaciens is a mixture of these three spiramycins, but it is also possible to obtain each of these spiramycins directly by fermentation, and in particular spiramycin I [see
L. Ninet and J. Verrier, Prod. Pharm. , 17 (4) 1 (1962)].



   The compounds of general formula (VII) in which the symbol
A represents a trans butenylene group and the symbol R1 is as defined above, and their non-toxic acid addition salts, can be prepared by partial reduction of spiramycins I, II and
III and their mixtures, and their non-toxic acid addition salts, by application or adaptation of the methods described above for the reduction of the compounds of general formula (I) in which the symbol A represents a butadienylene group (in which the double bonds are trans) and the symbols R1 and R2 are as defined above, for example as described in British patent No 785191.



   The term known methods as used in the present text means methods previously used or described in the chemical literature.



   The compounds of general formula (VII) and their non-toxic acid addition salts are antibiotics with antibacterial properties which make them extremely useful for the treatment of infections caused by Staphylococcus aureus and pneumococcal and streptococcal infections, and also, in veterinary medicine, for the treatment of mycoplasmosis (a disease also known as infectious sinusitis and infection of the air sacs) in turkeys, caused mainly by an organism of the type of those causing pleuropneumonia, known as name of Mycoplasma gatltsepttcurn.



   The compounds of general formula (VII) and their non-toxic acid addition salts also have a growth stimulating activity which makes them extremely useful for improving the growth rate and / or the rate of food transformation in mammals and birds, in particular those of domestic species, for example pigs, cattle, for example calves, and poultry, for example chickens and turkeys. This improvement in the growth rate of the animals means that they reach the desired weight, for example for marketing, in a shorter period of time than that which is usually necessary, or that they reach a higher weight within the same lapse of time. of time.



  The improvement in the rate of transformation of food by animals means that they consume less food to reach a given weight than similar animals which do not receive a compound of general formula (VII) or one of its non-toxic salts of addition of acid. When administered to poultry, the compounds of general formula (VII) are also useful for increasing egg production. The compounds of general formula (VII) are normally administered to mammals and birds in association with a balanced diet.



   The compounds of general formula (I) have a therapeutic activity and a growth-stimulating activity of type and degree similar to those of the compounds of general formula (VII) and, moreover, they have the important advantage of being less bitter than the compounds of general formula (VII), or even to be practically tasteless. This advantage is particularly important for the treatment of children and animals by oral administration of the compounds of general formula (I).



   Although the non-toxic acid addition salts of the compounds of general formula (I) may not have the advantage of being less bitter than the compounds of general formula (VII), they can be used for the purification of the compounds of general formula (I), for example by taking advantage of the differences in solubility between these salts and the bases from which they are derived, in water and in organic solvents, by techniques well known to specialists. The bases of general formula (I) can be regenerated from their salts by known methods, for example by treatment with a mineral base, for example an aqueous solution of sodium hydroxide.



   In British Patent No. 796311 are described hemiesters formed by spiramycins with dicarboxylic acids of the aliphatic series, hemiesters which are less bitter than the compounds of general formula (VII).



   However, the compounds of general formula (I) are even less bitter than the compounds described in this British patent
No 796311.



   The following compounds of general formula (I) and their mixtures are of particular interest:
Compound
A 2 'ethylsuccinate of spiramycin I;
B 2 'spiramycin ethylglutarate I;
C 2 'spiramycin methylsuccinate I;
D propylsuccinate-2 'of spiramycin I;
E 2 'spiramycin methylglutarate I;
F 2-spiramycin I methyl fumarate;
G 2-spiramycin I S-methylthiosuccinate-2 ';
H cyclohexylsuccinate-2 'of spiramycin I;
J spiramycin I ethylmalonate-2 ';
Spiramycin I phenylsuccinate-2 ';
L methyl- (RS) - (methyl-3) -succinate-2 'of spiramycin I;
M (2-phenylethyl) succinate-2 'of spiramycin I;
N tert.-butylsuccinate-2 'of spiramycin I;
O isopropylsuccinate-2 'of spiramycin I;
P ethylsuccinate-2 'of a mixture of spiramycins, and
Q dihydrochloride of compound A;
R dioxalate of compound A.



   The above compounds have been assigned the letters A to H and J to R for easy reference later in the text, for example in the tables.



   In experiments on therapeutic activity and biological activity in vitro, carried out on compounds of general formula (I), the following test results were obtained:
Test I - Mouse treated orally
 Mice are injected intraperitoneally with Staphylococcus aureus Smith suspended in mucin (0.5 ml). This infection represents approximately 100 times the minimum dose calculated to kill 100% of the mice. Each mouse is treated, orally, with doses of 16,31,62,5, 125 or 250 mg per kilo of body weight of the animal, of a compound of general formula (I). Each mouse is treated 1, 4, 24, 30 and 48 h after infection, with the dose chosen. The mice are observed for 7 d after infection.



   The results obtained from a compound of general formula (I) are compared with those obtained from spiramycin I used as standard compound.



   The results obtained are shown in Table I (b).



   The term DC50 signifies the curative dose calculated to cure 50% of the treated mice.



  Test 2 - Mouse treated subcutaneously
 Mice are infected intraperitoneally with Staphylococcus aureus Smith [approximately 100 times the minimum lethal dose for 100% of infected mice] suspended in mucin (0.5 ml).



  Each mouse is treated, subcutaneously, 1, 4, 24, 30 and 48 hours after infection, with doses of 1.6, 3.1, 6.25, 12.5 or 25 mg per kilo body weight of the animal, of a compound of general formula (I). The mice are observed for 7 d after infection.



   The results obtained from a compound of general formula (I) are compared with the results obtained from spiramycin
I used as a standard compound. Thus, compound A showed a CD50 of 17.5 mg per kilo of body weight of the animal, while spiramycin I showed a DC50 of 5.8 mg per kilo of body weight.

 

  Test 3 - In vitro activity
 For each compound to be tested, a range of concentrations is prepared in soy tryptone broth. Each test tube containing a different concentration, taken in this range, of test compound is inoculated with the organism chosen for the test:
Staphylococcus aureus (Oxford) or Escherichia coli B (Ennis) (approximately 105 organisms per tube) and, after incubation at 37oC, the lowest concentration of test compound, in said range, necessary to prevent the growth of the microorganism to testing is noted as the minimum inhibition concentration for this compound and this organism.



  The results obtained are shown in Table 1 (a).
EMI6.1


 <tb>



    <SEP> (a) <SEP> In <SEP> vitro. <SEP> Concentration <SEP> minimum Inhibition <SEP> <SEP> (b) <SEP> Mouse <SEP> DC50 <SEP> (channel <SEP> oral)
 <tb> <SEP> (ug / ml) <SEP> in <SEP> vîvo <SEP> (mg / kg)
 <tb> <SEP> S. <SEP> S.

   <SEP> aureus <SEP> Escherichia <SEP> coli <SEP> B <SEP> Spiramycin <SEP> I
 <tb> <SEP> Compose <SEP> (Oxford) <SEP> (Ennis)
 <tb> <SEP> A <SEP> 1.25 <SEP> 62 <SEP> 88 <SEP> 71
 <tb> <SEP> B <SEP> 1.25 <SEP> 62 <SEP> 135 <SEP> 71
 <tb> <SEP> C <SEP> 2.5 <SEP> 250 <SEP> 62 <SEP> 47
 <tb> <SEP> D <SEP> 2.5 <SEP> 250 <SEP> 90 <SEP> 47
 <tb> <SEP> E <SEP> 2.5 <SEP> 125 <SEP> 125 <SEP> 79
 <tb> <SEP> F <SEP> 2.5 <SEP> 250 <SEP> 125 <SEP> 62
 <tb> <SEP> G <SEP> 2.5 <SEP> 500 <SEP> 114 <SEP> 62
 <tb> <SEP> H <SEP> 2.5 <SEP> 250 <SEP> 67 <SEP> 41
 <tb> <SEP> J <SEP> 5 <SEP> 250 <SEP> 120 <SEP> 95
 <tb> <SEP> K <SEP> 2.5 <SEP> 250 <SEP> 76 <SEP> 41
 <tb> <SEP> L <SEP> 2.5 <SEP> 250 <SEP> 58 <SEP> 41
 <tb> <SEP> M <SEP> 2.5 <SEP> 250 <SEP> 64 <SEP> 41
 <tb> <SEP> N <SEP> 2.5 <SEP> 125 <SEP> 130 <SEP> 79
 <tb> <SEP> O <SEP> 1.25 <SEP> 125 <SEP> 96 <SEP> 62
 <tb> <SEP> P

    <SEP> 5 <SEP> 250 <SEP> 175 <SEP> 110
 <tb> Spiramycin <SEP> 0.62-2.5 <SEP> 62-250
 <tb>
 These results prove that the compounds of general formula (I) have an activity of the same order as that of the compounds of general formula (VII).



   The usefulness of the compounds of general formula (I) as antibiotics and growth stimulators is increased by the fact that they are relatively harmless to mammals, as demonstrated by the following test:
Toxicity to mice
 Mice are each treated orally with one of the compounds of general formula (I), and they are kept under observation for the next 3 d.



   The LD50 figures obtained (lethal doses for 50% of the treated mice) are shown in Table II, expressed in milligrams per kilo of body weight of the animal. None of the mice died during the experiment, even at doses of 1000 mg per kilogram of body weight.



   Table II
EMI6.2


 <tb> Compound <SEP> DL50
 <tb> <SEP> Au11000 <SEP>
 <tb> <SEP>> <SEP> 1000
 <tb>
 The examples which follow illustrate the preparation of the new compounds according to the present invention.



  Example 1:
Compound A
 A solution of ethyl chloroformyl-3-propionate (3.2 g) in anhydrous chloroform (50 ml) is added dropwise with stirring to a solution of spiramycin 1 (15 g) in anhydrous chloroform (100 ml). ). Maintaining a temperature of 0 to 10 "C throughout the addition, cooling in an ice bath. The mixture is left for 4 h at room temperature.



  The solvent is removed in vacuo and the residue is shaken with an aqueous solution of acetic acid (100 ml; 1N). The resulting mixture is extracted with diethyl ether (150 ml). The aqueous layer is separated and adjusted to about pH 7 by the addition of an aqueous solution of sodium hydroxide (1 ON). The mixture is then extracted with diethyl ether (150 ml) and the resulting ether extract is dried over magnesium sulfate. The diethyl ether is removed by distillation under reduced pressure to give 2 'ethylsuoeinate 2' of spiramycin 1 (4.5 g), in the form of a white solid, P.F. 100-110 C.



  Example 2:
Compound B
 Triethylamine (3 g) is added, with stirring, and, dropwise, a solution of ethyl chloroformyl-4 butyrate (5.1 g) in anhydrous chloroform (50 ml) to a solution of spiramycin I ( 24 g) in anhydrous chloroform (100 ml). The temperature is maintained at 0 to 10 ° C. for the entire duration of the addition, cooling in an ice bath. The mixture is left for 4 h at room temperature. The solvent is removed in vacuo and the residue is shaken with a aqueous acetic acid solution (100 ml; IN).

  The resulting mixture is extracted with diethyl ether (150 ml). The aqueous layer is separated and adjusted to about pH 7 by the addition of an aqueous solution of sodium hydroxide (1 ON). The mixture is then extracted with diethyl ether (150 ml) and the resulting ether extract is dried over magnesium sulfate. The diethyl ether is removed by distillation under reduced pressure to give 2 'ethylglutarate of spiramycin 1 (3.0 g), as a white solid, mp 87-95 C.



  Example 3:
Compound C
 Triethylamine (1.0 g) is added with stirring and a solution of methyl chloroformyl-3-propionate (1.7 g) in anhydrous chloroform (10 ml) is added dropwise to a solution of spiramycin I (8.4 g) in anhydrous chloroform (100 ml). The mixture is stirred for 3 h, then the solvent is removed in vacuo. To the resulting residue is added an aqueous solution of acetic acid (75 ml; 1N) and diethyl ether (100 ml). The mixture is shaken until the solid dissolves. The aqueous layer is separated and adjusted to about pH 7 by the addition of an aqueous solution of sodium hydroxide (1 ON).



  The mixture is then extracted with diethyl ether (100 ml) and the ether extract is dried over magnesium sulfate. The diethyl ether is removed by distillation under reduced pressure to give 2 'methylsuccinate of spiramycin 1 (1.0 g), as a white solid, mp 108-118'C.



  Example 4:
Compound D
 A solution of propyl chloroformyl-3-propionate (1.8 g) in anhydrous chloroform (10 ml) is added dropwise with stirring to a solution of spiramycin 1 (8.4 g) in anhydrous chloroform (70 ml). The mixture is stirred for a further 5 h, then the solvent is removed in vacuo. The residue is dissolved in an aqueous solution of acetic acid (100 ml; 1N) and the resulting mixture is extracted with diethyl ether (100 ml). The aqueous layer is separated and adjusted to about pH 7 by the addition of an aqueous solution of sodium hydroxide (1 ON). The mixture is extracted with diethyl ether (100 ml) and the resulting ether extract is dried over magnesium sulfate.

  The diethyl ether is removed by distillation under reduced pressure to give propylsuccinate-2 'of spiramycin 1 (0.7 g), in the form of a white solid, mp 88-96'C.



  Example 5:
Compound E
 Triethylamine (1.0 g) is added, with stirring, and, dropwise, a solution of methyl chloroformyl-4 butyrate (1.65 g) in anhydrous chloroform (20 ml) to a solution of spiramycin I (8.4 g) in anhydrous chloroform (70 ml). The mixture is stirred for a further 3 h and allowed to stand for 17 h. The reaction mixture is extracted with an aqueous solution of acetic acid (100 ml; 1N). The chloroform layer is separated and washed with an aqueous solution of sodium carbonate at 5% w / v (100 ml). The chloroform layer is dried over magnesium sulfate.

  The chloroform is removed by distillation under reduced pressure to give 2 'methylglutarate of spiramycin 1 (3.2 g), in the form of a buff-colored solid, mp 85-95 C.



  Example 6:
Compound F
 Sodium bicarbonate (8 g) and, dropwise, a solution of methyl chloroformyl-3 transacrylate (2.2 g) in anhydrous acetone (40 mil) are added to a solution of spiramycin, with stirring. 1 (8.4 g) in anhydrous acetone (80 ml). The mixture is stirred for a further 3 h and allowed to stand for 17 h, then the solid is filtered off. The filtrate is evaporated and the residue is dissolved in ethyl acetate (100 ml). This solution is extracted with an aqueous solution of acetic acid (75 ml; 1N). The aqueous layer is separated and extracted with chloroform (2 times 50 ml). The chloroform extracts are combined and washed with water (100 ml) and with an aqueous solution of sodium bicarbonate at 5% w / v (100 ml).

  The chloroform layer is then dried over magnesium sulfate. The chloroform is removed by distillation under reduced pressure, to give spiramycin 1-methyl-fumarate-2 '(2.7 g), m.p. 105-115 "C.



     Example 7 ..



  Compound G
 Triethylamine (1.1 g), ethyl chloroformate (1.1 g) are added with stirring and a solution of methanethiol (1.0 g) in anhydrous chloroform (50 ml) is added dropwise. ) to a solution of hemisuccinate-2 'of spiramycin 1 (9.4 g) in anhydrous chloroform (80 ml) cooled to 9-15 ° C. A temperature of -15 to -10 ° C. is maintained during the addition. the mixture is allowed to warm up to ambient temperature and is left there for 18 hours The solvent is removed by distillation under reduced pressure, diethyl ether (100 ml) is added to the residue and the insoluble materials are removed by filtration.

  The filtrate is extracted with an aqueous solution of acetic acid (100 ml; 1N). The aqueous layer is separated and adjusted to about pH 6 by the addition of an aqueous solution of sodium hydroxide (1 ON). The mixture is extracted with diethyl ether (100 ml), the ether extract is washed with a 5% w / v aqueous solution of sodium bicarbonate (100 ml) and dried over magnesium sulfate. The diethyl ether is removed by distillation under reduced pressure, to give 2 methylthiosuccinate-2 'of spiramycin 1 (1.4 g), m.p. 114-124 "C.



   The 2 'succinic hemiester of spiramycin I used in the preparation example above is prepared as follows:
 Succinic anhydride (6.0 g) is added with stirring to a solution of spiramycin I (50.4 g) in ethyl acetate (600 ml).



  The mixture is stirred for a further 3 h and is left for 18 h at room temperature. The solvent is removed in vacuo and diethyl ether (200 ml) is added to the residue. The solid is separated by filtration and dried, which gives the sought hemiester (38.6 g),
P.F. 140-148 C.



  Example 8:
Compound H
 Sodium bicarbonate (4.0 g) is added with stirring and a solution of 3-cyclohexyl chloroformyl propionate (2.4 g) in anhydrous acetone (10 ml) is added dropwise to a solution. of spiramycin 1 (8.4 g) in anhydrous acetone (70 ml). The mixture is stirred for 3 h, then allowed to stand for 16 h. This mixture is filtered and the filtrate is evaporated under reduced pressure. The residue is dissolved in ethyl acetate (70 ml) and the resulting mixture is extracted with an aqueous solution of acetic acid (75 ml; 0.5N). The aqueous layer is separated and extracted with chloroform (2 times 50 ml).

  The chloroform extracts are combined, washed with water (100 ml) and with an aqueous solution of sodium bicarbonate at 5% w / v (75 ml), and dried over magnesium sulfate.



  The chloroform is evaporated in vacuo, which gives cyclohexylsuccinate-2 'of spiramycin 1 (3.4 g), m.p. 90-100 "C.



  Example 9:
Compound J
 Sodium bicarbonate (8 g) and, dropwise, a solution of 2-chloroformyl ethyl acetate (1.5 g) in anhydrous acetone (20 ml) are added with stirring to a solution. of spiramycin 1 (8.4 g) in anhydrous acetone (75 ml). Stirring is continued for 5 h, then the mixture is left to stand for 17 h. The solid is removed by filtration and the filtrate is evaporated in vacuo. The residue is dissolved in ethyl acetate (100 ml) and this solution is extracted with an aqueous solution of acetic acid (75 ml; 0.5N).

  The aqueous layer is separated and extracted with chloroform (2 times 50 ml). The chloroform extracts are combined, washed with water (100 ml) and with an aqueous solution of sodium bicarbonate at 5% w / v (100 ml), and dried over magnesium sulfate. The chloroform is removed by distillation under reduced pressure to give 2 'ethylmalonate of spiramycin 1 (2.0 g), m.p. 89-99 "C.



   Example 10:
 Compound K
 Triethylamine (1.5 g) and chlorofor are added with stirring
 ethyl miate (1.7 g) to a solution of phenyl acid succinate
 (2.7 g) in anhydrous chloroform (50 ml), cooled to -15 C.
 dropwise a solution of
 spiramycin 1 (8.4 g) in anhydrous chloroform (30 ml), while maintaining the temperature of the mixture at - 15 to - C. The mixture is allowed to warm up to room temperature and is left there for 5 p.m.

  This mixture is evaporated in vacuo, and to the residue is added ethyl acetate (75 ml) and an aqueous solution of sodium bicarbonate at 5% w / v (75 ml). The organic layer is separated and extracted with an aqueous solution of acetic acid (75 ml; 0.5N). The aqueous layer is then extracted with chloroform (2 times 50 ml). The chloroform extracts are combined, washed with water (100 ml) and with an aqueous solution of sodium bicarbonate at 5% w / v (75 ml) and dried over magnesium sulfate. By evaporation of the chloroform under reduced pressure, phenylsuccinate-2 'of spiramycin I (5.1 g), P.F. 105-115 "C.



  Example ll:
Compound L
 Sodium bicarbonate (8 g) is added, with stirring, and, dropwise, a solution of (RS) -methyl-3-chloroformyl-3-methyl-propionate (2.5 g) in anhydrous acetone (20 mi) to a solution of spiramycin 1 (8.4 g) in anhydrous acetone (80 ml). Stirring is continued for 4 h, then the mixture is left to stand for 18 h.



  The solid is removed by filtration and the filtrate is evaporated in vacuo.



  The residue is dissolved in an aqueous solution of acetic acid (75 ml; 0.5N). The resulting mixture is extracted with ethyl acetate (2 times 50 ml) then with chloroform (2 times 50 ml). The chloroform extracts are combined, washed with water (100 ml) and with an aqueous solution of sodium bicarbonate at 5% w / v (100 ml) and dried over magnesium sulfate.



  The chloroform is removed by distillation under reduced pressure, to give methyl- (RS) - (methyl-3) -succinate-2 'of spiramycin I (1.7 g), m.p. 92-102 "C.



  Example 12:
Compound M
 Triethylamine (1.1 g) and ethyl chloroformate (1.1 g) are added with stirring to a solution of spiramycin 1-2 'hemisuccinate (9.4 g) in anhydrous chloroform ( 80 ml), cooled to - i 5 C. A solution of 2-phenyl ethanol (1.8 g) in anhydrous chloroform (20 ml) is added dropwise, while maintaining the temperature at - 15 to - C. The mixture is then allowed to warm to room temperature and left there for 18 h.

  The solvent is removed under reduced pressure and the residue is dissolved in an aqueous acetic acid solution (100 ml; 1N). The solution is extracted with diethyl ether (100 ml). The aqueous layer is separated and adjusted to about pH 6 with an aqueous solution of sodium hydroxide (ION). The mixture is extracted with diethyl ether (100 ml), the resulting ether extract is washed with an aqueous solution of sodium bicarbonate 5% w / v (100 ml) and dried over magnesium sulfate .



  The diethyl ether is removed by distillation under reduced pressure, to give spiramycin 1 (2-phenylethyl) succinate-2 '(2.3 g), m.p. 58-68 C.



  Example 13:
Compound N
 Triethylamine (1.1 g) and ethyl chloroformate (1.1 g) are added, with stirring, to a solution of spiramicin 1-2 'hemisuccinate (9.4 g) in anhydrous chloroform ( 80 ml), cooled to -15 "C. Then added dropwise to the resulting mixture
 solution of tertiary butanol (1.2 ml) in anhydrous chloroform
 (40 ml), while maintaining the temperature between - 15 and - i OC. We
 continue to stir while allowing the reaction mixture to warm to room temperature, then allow the mixture to stand
 for 6 p.m.

  The solvent is removed under reduced pressure and
 dissolves the residue in an aqueous solution of acetic acid (100 ml;
   IN). The mixture is extracted with diethyl ether (100 ml). We
 separates the aqueous layer and is adjusted to about pH 6 with an aqueous solution of sodium hydroxide (ION). We extract the
 mixture with diethyl ether (100 ml), the ether extract is washed
 resulting in a 5% aqueous sodium bicarbonate solution
 w / v (100 ml), and dried over magnesium sulfate.



   The diethyl ether is removed by distillation under reduced pressure, to give tert.-butylsuccinate-2 'of spiramycin 1 (0.9 g), P.F.



      99-108 C.



  Example 14:
 Compound O
 A solution of triethylamine (1.0 g) in anhydrous chloroform (10 ml) is added, with stirring, and, dropwise, a solution of isopropyl-3-chloroformyl propionate (2.2 g) in anhydrous chloroform (20 mil) to a solution of spiramycin 1 (8.4 g) in anhydrous chloroform (75 ml). The mixture is stirred for 5 h, then allowed to stand for 18 h.

  The solvent is removed under reduced pressure, and, with shaking, an aqueous solution of acetic acid (100 ml; IN) and diethyl ether (100 ml) are added. The aqueous layer is separated and adjusted to about pH 6 with an aqueous solution of sodium hydroxide (ION). The mixture is extracted with diethyl ether (100 ml), the resulting ether extract is washed with an aqueous solution of sodium bicarbonate 5% w / v (100 ml) and dried over magnesium sulfate . The diethyl ether is removed under reduced pressure, to give isopropylsuccinate-2 'of spiramycin 1 (1.2 g), mp 105-115C.



  Example 15:
Compound P
 Sodium bicarbonate (8 g) and, dropwise, a solution of ethyl chloroformyl-3-propionate (2.5 g) in anhydrous acetone (30 ml) are added with stirring to a solution of '' a mixture of spiramycins (8.4 g) in anhydrous acetone (80 ml). The mixture is stirred for 5 h and then allowed to stand for 18 h. The solid is removed by filtration and the filtrate is evaporated under reduced pressure.

  The residue is dissolved in an aqueous acetic acid solution (75 ml; 0.5N) and the resulting mixture is extracted with ethyl acetate (2 times 50 ml), then with chloroform (2 times 50 ml ). The chloroform extracts are combined, washed with water (100 ml) and with an aqueous solution of sodium bicarbonate at 5% w / v (100 ml) and dried over magnesium sulfate.



  The chloroform is removed under reduced pressure, which gives 2 'ethylsuccinate from the mixture of spiramycins (4.4 g), m.p. 80-90 C.



   The mixture of spiramycins used in the preparation example above comprises approximately, by weight, 60% of spiramycin I, 20% of spiramycin II and 20% of spiramycin III.

 

  Example 16:
Compound Q
 A solution of hydrogen chloride in ethanol is added dropwise to a solution of 2 'ethylsuccinate of spiramycin I (1.5 g) in diethyl ether (50 ml) until 'it no longer precipitates solid. The white solid is separated by filtration, washed with diethyl ether and dried, to give the spiramycin 1 ethylsuoeinate-2 'dihydrochloride (1.4 g), PF 64-74 "C .



  Example 17:
Compose
 A saturated solution of oxalic acid in diethyl ether is added dropwise to a solution of 2 'ethylsuccinate of spiramycin 1 (1.5 g) in diethyl ether (50 ml) until that it does not precipitate any more solid. The white solid is separated by filtration, washed with diethyl ether and dried, which gives the dioxalate of 2-ethylsuccinate-2 'of spiramycin 1 (1.6 g), m.p.



     85-97 C.



   The present invention encompasses in its field pharmaceutical compositions which comprise at least one compound of general formula (I), in association with a compatible and pharmaceutically acceptable support or coating. The compounds of general formula (I) will normally be administered vaginally, rectally, parenterally or, more especially, orally.



   Solid compositions for oral administration include tablets, pills, dispersible powders and granules. In these solid compositions, one or more of the active compounds are mixed with at least one inert diluent such as mannitol, calcium carbonate, potato starch, alginic acid or lactose. These compositions can also comprise, as is common practice, additional substances other than inert diluents, for example lubricants, such as magnesium stearate. Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs, containing the inert diluents usually used in pharmacy, such as water and paraffin oil.

  In addition to the inert diluents, these compositions may also include adjuvants, such as wetting agents and suspending agents, sweetening, flavoring agents, perfumes and preservatives. The compositions according to the invention for oral administration also include capsules of absorbable substances, such as gelatin, containing one or more of the active substances with or without the addition of diluents or excipients.



   The compositions for oral administration include granules, prepared in a manner known per se, which means in a manner previously used or described in the literature, and which can be dispersed in water before use.



   Solid compositions for vaginal administration include ova prepared in a manner known per se and containing one or more of the active compounds.



   Solid compositions for rectal administration include suppositories prepared in a manner known per se and containing one or more of the active compounds.



   The preparations according to the invention for parenteral administration include aqueous or non-aqueous sterile solutions, suspensions or emulsions. As solvents or non-aqueous suspension media, there may be mentioned propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. These compositions can also include adjuvants such as preservatives, wetting agents, emulsifiers and dispersants. These compositions can be sterilized, for example by filtration through a bacteria-retaining filter, by incorporation of sterilizing agents, by irradiation or by heating. They can also be manufactured in the form of sterile solid compositions, which can be dissolved extemporaneously in sterile water or another sterile injectable medium.



   The percentage of active ingredient in the compositions according to the invention is variable, but it is necessary that it represents a proportion such that a dosage suitable for the desired therapeutic effect is obtained. Of course, several dosage forms can be administered at approximately the same time. In general, preparations should normally contain at least 0.025% by weight of active substance when they are intended for administration by injection; for oral administration, preparations will normally contain at least 0.1% by weight of active substance. The dose used depends on the desired therapeutic effect, the method of administration and the duration of treatment.

  In adult humans, the doses can range from 0.1 to 10 g / d, but are generally from 1 to 4 g / d, by oral administration for the treatment of bacterial infections.



   The following examples illustrate the pharmaceutical compositions according to the invention.



  Example 18:
 Tablets having the following composition are prepared by the usual technique:
 Spiramycin I ethylglutarate-2 '0.250 g
 starch 0.150g
 colloidal silica 0.070 g
 magnesium stearate 0.030 g
Example 19:
 Granules having the following composition are prepared according to the usual technique:
 2% spiramycin I ethylglutarate 5% by weight
 mannitol 95% by weight


    

Claims (10)

 CLAIMS 1. Spiramycin derivatives, of general formula: EMI1.1  in which R1 represents an acetyl or propionyl group or a hydrogen atom, A represents a butenylene or butadienylene group, any double bond present in group A being trans, and R2 represents a group of general formula: : EMI1.2  in which Z represents a bivalent alkylene group, saturated or unsaturated, straight or branched chain, containing from 1 to 6 carbon atoms, Y represents a sulfur or oxygen atom, and R3 represents a straight chain alkyl group or branched containing from I to 6 carbon atoms, a cycloalkyl group containing from 3 to 8 carbon atoms, an aryl group or an aralkyl group in which the alkylene residue is straight or branched chain and contains from I to 6 carbon atoms, and their acid addition salts.  2. As a compound according to claim 1:  Spiramycin I 2 'ethylsuccinate;  Spiramycin I ethylglutarate-2 ';  spiramycin I methylsuccinate-2 ';  spiramycin I propylsuccinate-2 ';  spiramycin I methylglutarate-2 ';  spiramycin I 2 'methyl fumarate-2';  spiramycin I S-methylthiosuccinate-2 ';  spiramycin I cyclohexylsuceinate-2 ';  Spiramycin I ethylmalonate-2 ';  spiramycin I phenylsuccinate-2 ';  spiramycin I methyl- (RS) - (methyl-3) -succinate-2 ';  (2-phenylethyl) sueeinate-2 'of spiramycin I;  spiramycin I tert.-butylsuccinate-2 ';    Isopropylsueeinate-2 'of spiramycin I;  spiramycin I ethylsuccinate. 2 'dihydrochloride; ;  spiramycin I ethylsuccinate-2 'dioxalate  3. Process for the preparation of the compounds of the general formula (I) according to claim 1 and in which the symbols R1, A and R2 are as defined in claim 1 and their acid addition salts, characterized in that that compounds of general formula are reacted: EMI1.3  wherein R 'and A are as defined in claim 1, and R4 represents a group of general formula: EMI1.4  in which Z is as defined in claim 1, with compounds of general formula:  R3YH (V) in which Y and R3 are as defined in claim 1, in the presence of a compound of general formula: :  X1COOR5 (VI) in which X1 represents a bromine or chlorine atom and R5 represents an alkyl group containing from 1 to 4 carbon atoms and an acid acceptor, at a temperature of - 30 to + 30 C and in presence of an anhydrous inert organic solvent, and, where appropriate, the compound obtained is converted into one of its salts by reaction with an acid.  4. Process for the preparation of the compounds of the general formula (I) according to claim 1 and in which the symbols Rt, A and R2 are as defined in claim 1 and their acid addition salts, characterized in that that compounds of general formula are reacted: EMI2.1  in which R1 and A are as defined in claim 1, with compounds of general formula:  X2OCZCOYR3 (VIII) in which X2 represents a bromine or chlorine atom or a group of general formula: :  -OCOOR5 (IX) in which R5 is as defined in claim 3 and Z, Y and R3 are as defined in claim 1, in the presence of an acid acceptor or of a mineral base, at a temperature of -30 to +30 "C and in the presence of an anhydrous inert organic solvent, and, where appropriate, the compound obtained is converted into one of its salts by reaction with an acid.  5. Process for the preparation of the compounds of general formula (I), in which the symbol A represents a trans butenylene group and the symbols R 'and R2 are as defined in claim 1, characterized in that a compound of general formula (I) in the formula of which the symbol A represents a butadienylene group, in which the double bonds are trans, and the symbols R1 and R2 are as defined in claim 1, by the method according to claim 3, and that it is partially reduced by catalytic hydrogenation.  6. Method according to claim 5, characterized in that a noble metal or one of its oxides is used as catalyst, either alone or deposited on alumina, charcoal or barium sulphate.  7. Process for the preparation of the compounds of general formula I in which the symbol A represents a trans butenylene group and the symbols Rl and R2 are as defined in claim 1, characterized in that a compound of general formula is prepared (I) in which the symbol A represents a butadienylene group, in which the double bonds are trans, and the symbols R1 and R2 are as defined in claim 1, by the method according to claim 4, and that it is reduced partially by catalytic hydrogenation.  8. Method according to claim 7, characterized in that a noble metal or one of its oxides is used as catalyst, either alone or deposited on alumina, charcoal or barium sulphate.  9. Pharmaceutical composition comprising, as active ingredient, at least one spiramycin derivative according to claim 1, or one of its non-toxic acid addition salts, in association with a compatible and pharmaceutically compatible support or coating. acceptable.  10. Composition according to claim 9, characterized in that it comprises ethylsuccinate-2 'of a mixture of spiramycins containing approximately, by weight, 60% of spiramycin I, 20% of spiramicin II and 20% of spiramicin III .  The present invention relates to novel antibiotics, methods of preparing them and pharmaceutical compositions containing them.  The present invention relates to new spiramycin derivatives of general formula:    (Formula at the head of the following page) in which R1 represents an acetyl or propionyl group or, preferably, a hydrogen atom, A represents a butene group or, preferably, butadienylene group (all the double bonds present in the group A being trans), and R2 represents a group of general formula: : EMI2.2  in which Z represents a bivalent alkylene group, saturated or unsaturated, straight or branched chain, containing from 1 to 6 carbon atoms (for example an ethylene group), Y represents a sulfur atom or, preferably, a d atom oxygen, and R3 represents a straight or branched chain alkyl group containing from 1 to 6 carbon atoms, preferably from 1 to 3, a cycloalkyl group containing from 3 to 8 carbon atoms, an aryl group (for example phenyl) or an aralkyl group (for example phenylalkyl) in which the alkylene residue is straight or branched chain and contains from 1 to 6 carbon atoms, and their mixtures,  especially mixtures in which the compounds of general formula (I) contained therein differ only with regard to the symbol R1 and are identical with regard to the symbols A and R2, as well as the non-toxic addition salts acid of these compounds.  The spiramycin derivatives of general formula (I) in the formula of which R2 represents a group of general formula (II) in which Z represents an ethylene or trimethylene group, Y ** ATTENTION ** end of the CLMS field may contain start of DESC **.