CS199258B2 - Process for preparing new antibiotic compounds of rifamycine-s,substituted in 3 position by aliphatic substituted aminogroup - Google Patents

Abstract

PROCESS FOR THE MANUFACTURE OF NEW ANTIBIOTICALLY ACTIVE COMPOUNDS The invention refers to a process for the manufacture of selected 3-amino-rifamycin S and -SV derivatives, the amino group being of the aliphatic type, and more precisely being a 4R-1-piperazinyl- radical which is optionally substituted at the C-atoms by lower alkyl groups, and wherein R represents a hydrocarbon radical of the formula (III) , wherein Z1 - Z3 represent various hydrocarbon groups, including aromatic and cycloaliphatic groups, and Z' and Z" are optionally hydrogen or lower alkyl groups. These new compounds possess beyond a high general antibiotic action an especially high anti - tuberculosis effect, and are distinguished by a low toxicity.

Description

It is an object of the present invention to provide novel rifamicin-S and -VV derivatives with high antibiotic activity. They are compounds of rifamycin-S (1) substituted in the 3-position by an aliphatic substituted amino group of the formula I

3f

wherein in formula I

A represents a 4R-1-piperazinyl radical, optionally substituted on carbon atoms with C1-C7 alkyl groups in which

R is a hydrocarbon radical of formula III

and their respective hydroquinones (compounds II), i.e. rifamycin SV derivatives, which, instead of the naphthoquinone ring, contains a naphthohydroquinone ring of the sub-formula wherein n = 0 or 1,

Z 1 represents an alkyl radical having from 1 to 7 carbon atoms or an alkenyl radical having from 2 to 7 carbon atoms which may be substituted by unsubstituted or alkyl groups having from 1 to 7 carbon atoms substituted by phenyl, cycloalkyl or cycloalkenyl, or phenyl or unsubstituted; a C 1 -C 7 alkyl group substituted by a cycloalkyl or cycloalkenyl radical, wherein the phenyl radicals may also be substituted by halogens,

7.2 is hydrogen or an alkyl radical of 1 to 7 carbon atoms or an alkenyl radical of 2 to 7 carbon atoms, and

Z3 is C1-C7alkyl or C2-C7alkenyl, or optionally together with " Z " is a double bond, and Z2 and Z3 together are C1-C7alkylidene or C2-C7alkylidene. up to 7 carbon atoms and Z 1 and Z 3 together represent an alkylene or alkenylene residue, but Z 1 + Z 2 may also represent such an alkylene or alkenylene residue when Z 3 with "Z" represents a double bond, and each of Z 1 and Z 1 represents either hydrogen or an alkyl group having 1 to 7 carbon atoms, wherein in said groups or cycloaliphatic groups formed by an alkylene or alkenylene group Z 1 + Z 3 or Z 1 + Z 2, one or more non-adjacent ring atoms C-C in the ring may optionally be connected directly or indirectly to each other an alkylidene group having 1 to 7 carbon atoms or an alkylene group, or one carbon atom of an alkylene group it may be spirocyclically substituted and these groups have at most 12 carbon atoms, or R represents an allyl group, and their salts and quaternary ammonium salts.

Alkenyl, cycloalkenyl, alkenylidene or alkenylene groups are to be understood as meaning hydrocarbon radicals having at least one double bond.

Lower alkyl, lower alkenyl and lower alkylidene or lower alkenylidene groups are groups of up to 7 carbon atoms, in particular 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, straight or branched, butyl, pentyl bonded in any position , hexyl or heptyl groups, vinyl, allyl or methallyl, 1-propenyl, methylene, ethylidene, propylidene, butylidene, isopropylidene or isobutylidene groups. The alkylene or alkenylene group Z 1 + Z 3 together with the carbon atom to which it is attached form a cycloalkyl or cycloalkenyl group having a total of at most 12 carbon atoms, again being a cycloalkenyl group having at least one double bond. Preferred are straight or branched alkylene groups which thus form cycloalkyl radicals having 3 to 8 ring carbon atoms and in particular 5 to 6 ring carbon atoms or cycloalkenyl radicals having preferably 5 to H8 ring carbon atoms. Alkenylene groups or straight chain alkylene groups together with said carbon atom of the above formula form unsubstituted cycloaliphatic rings; in the case of branched alkylene groups, the rings formed are substituted with alkyl groups, in particular lower alkyl groups of 1 to 4 carbon atoms, primarily methyl groups . Such cycloalkyl groups having 3 to 8 ring carbon atoms and ^r are for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 4-methyl-cyclohexyl, 2,6-dimethylcyclohexyl, 3,5-dimethyIcyklohexylové groups or l-Methyl-44sopropylcyklohexylové (p 1-methyl-3-isopropylcyclohexyl)

(m-menthyl) groups.

Cycloalkenyl groups include, for example:

1-cyclopenten-1-yl

2-cyclopenten-1-yl,

3-cyclopenten-1-yl, 2,4-cyclopentadiene-1-yl,

1-cyclohexen-1-yl,

2-cyclohexen-1-yl,

3-cyclohexen-1-yl,

1,3-cyclohexadien-1-yl a

1,4-cyclohexadien-1-yl,

1- p -Menthen-4-yl

2- p-Menthen-1-yl

3- p-menthen-1-yl or, optionally, other isomers of this group, or the corresponding compounds with two double bonds, such as the hydrocarbon radicals of terpinines, fellandrenes, limonenes or menthadienes.

A cycloalkyl or cycloalkenyl group, optionally or occurring in or together with Z1 + Z2, may be unsubstituted or substituted by lower alkyl groups and contain at most 12 carbon atoms, and in particular is unsubstituted or lower alkyl groups, in particular alkyl groups (C až-C,) groups, in particular methyl / C skupin-C cyk-substituted cycloalkyl radical, in particular C až-C 6 6 ring, or C 5-C cyk-C cyk-cycloalkenyl radical and may be for example, one of the specific groups just mentioned.

19 '' 2 * N 8 i 5

Cycloalkyl - or - cycloaikenyl - groups may also be linked - via a bridge directly or through a lower alkylidene or alkylene - group, preferably - from 1 to 4 - carbon atoms, such as methylene, ethylene - or isopropylidene, or may be spirocyclically-substituted, i. may - also form a bicyclic polycyclic particular - ^are systems that are derived from or polycykloalkanů - bicycloalkanes - - or -spirocykloalkanů and their unsaturated derivatives ·. . Such groups are, for example, hydrocarbon residues of bicyclohexanes, -bicycloheptanes or bicyclooctanes and their derivatives substituted with lower alkyl groups, such as hydrocarbon residues of bicyclic terpenes - thujane, pinane - or bornane groups.

The aforementioned phenyl residues may be unsubstituted - or -. -substituted by - halogen, such as chlorine, fluorine or bromine, or by - lower alkyl radicals, i.e. radicals having from 1 to 7 carbon atoms, in particular by methyl groups.

The group --R preferably - has - at most. 35 carbon atoms, in particular -4 to 16 carbon atoms.

The lower -alkyl groups Z 'or - Z' - in - the above formula III - are the alkyl groups - - s. - - to - 7 - carbon atoms, - especially with 1 to 4 carbon atoms, as one of the above-mentioned - groups,. . especially methyl groups.

Of the compounds (I) and (II), the following are of particular interest - those in which the A group - represents a 4R-1-piperazinyl moiety - unsubstituted on carbon atoms, where R has one of the formulas below:

8 is substituted by C 4-C p alkyl, p _γ represents such a -cycloalkyl- or cycloalkenyl or phenyl group. -group, - like ..- is -. as defined above - phenyl, and alk is - straight - or branched alkylene - with 2 - to - 4 atoms. a carbon - or - alkylidene - group. : s - 1 -. up to -4 carbon atoms, and - their. derivatives, - - which - - - on - one. or - both carbon atoms - (CH 2) are substituted by a methyl group. The alkyl, alkenyl, alkylene, or alkenylidene groups having from 1 to 7 carbon atoms are, for example, the - above - said groups having, in particular, 1 to 4 carbon atoms, and are, in particular, methyl, ethyl or propyl groups, optionally vinyl or allyl groups; - or - methallyl - groups, - optionally - methylene, - ethylidene, propylidene - or p-propylidene - groups, optionally vinylidene - - or - 2-propene-ylidene groups. 2 - to -4 carbon - atoms - is - in particular - it trimethylene ethylene - group. - cycloalkyl - or - ^{cycloalkenyl:.} a group -. in - - the substituents R - - c _y - is - in particular. such - group - - with 5 - to - 6 atoms - - carbon - in ¹ ring, - hence cyclopentyl not - a cyclohexyl group, or, respectively, a hydrocarbon residue containing one unsaturated one. bond - and / or substituted by lower-alkyl groups such as - for example. above. groups. Preferably, 1 - to 3 - of said phenyl groups or - are optionally present in said phenyl - groups - or - cycloalkyl rings. cyclobytes, as described above, are linked by a bridge, and are then the residues of the aforementioned type, for example, the residues of said bicyclic terpenes.

Of interest are - i - derivatives of the just-described compounds - sub-formulas - IV - to - VII, - where - V3 stands for - alkyl - or. an alkenyl group having from 1 to 7, for example from 1 to 4 carbon atoms.

Also preferred are a group of (I) -a (II) compounds of which A is 4R-1-piperazinyl, unsubstituted on carbon, wherein R is below - of formula VIII iz iz - Ш, --x, I -1-¾ p Щ (V //))

wherein - n - 1 - or - 2 and - each - of the substituents VI and V 2 is an alkyl - or - alkenyl group having 1 to 7 carbon atoms,

V3 - represents - - hydrogen or together - with - V-2-alkylidene or -alkenylidene-group having 1 to 7 carbon atoms, ph represents - unsubstituted. or - chlorine or bromine atoms - and / or - methyl - substituted - phenyl, cy - means -. cycloalkyl. or a -cycloalkenyl-group having 5 to 8 carbon atoms in the ring, the ring optionally being where η = 1 or 2 and m = 1 to 4,

P is hydrogen or (C 1 -C 4) alkyl;

W 1, W 2 and W 3 denote alkyl and / or alkenyl groups having up to 4 carbon atoms, optionally present on any ring carbon atom, their unsaturated derivatives and derivatives of these compounds with a cycloaliphatic ring bridge of formula VIII, especially those wherein two non-adjacent carbon atoms are endocyclically bonded together by a direct bond C, C or via a lower alkylidene or alkylene group having 1 or 2 to 4 carbon atoms, and / or derivatives thereof, which are substituted on one or both carbon atoms of the group [ (Methyl). Examples of saturated and unsaturated cycloaliphatic rings of formula VIII can be found among those generally described above for compounds (I) and (II), which implies that, in the case of unsaturated rings, double bonds in the 1 or 2 position are particularly suitable. alkyl or alkenyl groups as well as groups forming a bridge of the bicyclic or polycyclic radicals, in particular the groups mentioned above for the similar cycloaliphatic _C.

In particular, m = 1 or 2 in these compounds. In particular, when substituted, 1 to 2 methyl groups are attached to these rings.

Of the compounds according to the invention of sub-formulas III to VIII of the radical R as defined above, those compounds in which n = 1, and in particular those in which the radicals Vi to V3, py, cy and alk in the formula VII have the as defined above, optionally in formula VIII, P is hydrogen and the cycloaliphatic ring is a cyclopentyl or cyclohexyl ring or a corresponding moiety containing one unsaturated bond and / or substituted with methyl groups, in particular 1 to 2 methyl groups.

Among the novel compounds (I) and (II) are, for example

3- (4-isobutyl-1-piperazinyl) -rifamycin S and SV,

3- (4- (2-ethylbutyl) -1-piperazinyl} -rifamycin S and SV,

3- [4- (2-methylbutyl) -1-piperazinyl] -rifamycin S and SV,

3- (4- (2-methylpentyl) -1-piperazinyl] -rifamycin S and SV,

3- (4- (2-phenylpropyl) -1-piperazinyl] -rifamycin S and SV,

3- (4-Methallyl-1-piperazinyl) -rifamycin

S and SV,

3- (4- (2-Methyl-3-butenyl) -1-piperazinyl] -rifamycin S and SV,

3- (4-cyclohexylmethyl-1-piperazinyl) -rifamycin S and SV,

3- (4- (4-methylcyclohexylmethyl) -1-piperazinyl] -rifamycin S and SV,

3- (4- (3-tert-butylcyclohexylmethyl) -1-piperazinyl) -rifamycin S and SV,

3- (4- (4-isobutylcyclohexylmethyl) -1-piperazinyl] -rifamycin S and SV,

3- (4- (4-isopropylcyclohexylmethyl) -1-piperazinyl] -rifamycin S and SV,

3- [4- (3-methylcyclohexylmethyl) -1-piperazinyl] -rifamycin S and SV,

3- (4- (3,4-dimethylcyclohexylmethyl) -1-piperazinyl] -4-rifamycin S and SV,

3- [4- (3,5-dimethylcyclohexylmethyl) -1-piperazinyl] -rifamycin S and SV,

3- (4- (3-cyclohexenylmethyl) -1-piperazinyl] -rifamycin S and SV, v

3- (4-cyclopropylmethyl-1-piperazinyl) -rifamycin S and SV,

3- (4-cycloheptylmethyl-1-piperazinyl) -rifamycin S and SV,

3- (4- (2-cyclohexylpropyl) -1-piperazinyl] -rifamycin S and SV;

3- (4- (2-cyclohexylbutyl-1-plperazinyl) -rifamycin S and SV,

3- (4-cyclooctylmethyl-1-piperazinyl) -rifamycin S and SV,

3- (4- (2-methylcyclohexylmethyl) -1-piperazinyl] -rifamycin S and SV,

3 - ((bicyclo-2,2,1) hex-2-ylmethyl) -1-piperazinyl / rifamycin S or SV.

Derivatives of 3-aminorifamycin S and SVsantibiotic activity are already known. Thus, French patent specification no.

No. 490,183 discloses rifamycin S derivatives

SVs which are substituted at the 3-position with an amino group of aliphatic nature. These compounds have high; they have an antibiotic effect and in particular have antituberculous activity. The compounds according to the invention then have a higher antituberculous effect compared to the compounds described in the French patent and other known rifamycin derivatives with antituberculous activity, as can be demonstrated in animal experiments, for example mice. Thus, when administered orally to mice infected with Mycobacterium bovis, these compounds exhibit a marked tuberculostatic effect at doses ranging from 1 mg / kg to 40 mg / kg. For example, 3- [44butyl-1-piperazinyl) riflamycin SV has an ED 50 value of 1 mg / kg in this assay. At the same time; 3- (4-Cyclohexylmethyl-1: -piperazinyl) rif amycin SV also shows an ED59 = 1 mg / kg. In addition, the compounds produced by the method ¹ according to the invention are distinguished by a very wide therapeutic range, since the more pronounced toxicity occurs only at very high doses. Thus, for example, the oral ILD50 of both of the above specific compounds is greater than 500 mg / kg.

The high antituberculous activity of these compounds can also be demonstrated by in vitro experiments. Thus, the minimum inhibitory concentration in vitro against Mycobacterium bovis of the aforementioned 344butyl-1-piperazinyl) rifamycin SV is 30 times lower than that of the known tuberculostatic rifamicin drug rifampicin, i.e. 3-4 (methyl-1-piperazinyl-iminomethyl) rifamycin SV .

These compounds also have good antibacterial activity, as can also be demonstrated in animal experiments such as mice. Thus, they exhibit marked antibacterial efficacy in mice infected with staphylococci at oral doses of 0.2 to 40 mg / kg.

The novel compounds can therefore be used as medicaments, primarily for tuberculous infections, but also for other infections, such as leprosy, or those caused by pyogenic germs, such as staphylococci. However, the novel compounds are also valuable intermediates for the production of other useful substances, in particular for the production of pharmacologically active compounds.

The novel compounds according to the invention can be obtained in a manner known per se, for example according to the data contained in the aforementioned French patent, i.e. by reacting rifamycin S with an amine corresponding to the group to be introduced at the 3-position.

For example, according to another method, an unsubstituted - or carbon-substituted pieprazinyl residue 'is introduced into the 3-position of rifamycin S or SV as described above, followed by introduction of the residue R by reaction with an alkylating agent into the N' position of the piperazinyl residue.

The process for the preparation of the compounds 1 and 11 described in this application is characterized in that rifamycin S is reacted with an amine of the formula HA, where A is as defined above, in an organic solvent free of hydroxyl groups, preferably of low polarity, especially in dioxane, or even without solvent, using an excess of the amine, at temperatures ranging from room temperature to 100 ° C, or with 3- (1-piperazinyl) -rifamycin S or SV, which may optionally be substituted with alkyl radicals of 1 to 7 carbon atoms on the piperazinyl radical, reacted with an alkylating agent suitable for the N-alkylation of the piperazines of the general formula XR, where X is a halogen atom or a residue ^, sulfonic acids or a acidic inorganic acid residue, in the presence of bases at temperatures ranging from room temperature to 100 ° C using 1 mole each of the reaction reagents in an internal solvent; Such as any chlorinated aliphatic hydrocarbon or lower 'aliphatic' alkanol, through which alkylating agent introduces the radical R, wherein R is as defined above, and the obtained 3-amino-4-substitution product rifamycin S or rifamycin SV is to be and / or if desired, the hydroquinone obtained before or after isolation is oxidized to quinone, or the obtained hydroquinone is reduced to hydroquinone and / or the compound obtained is converted into a salt.

The reaction of rifamycin S with an amine-HA ¹ * is conveniently carried out in an organic solvent free of hydroxyl groups of preferably low polarity such as halogenated aliphatic hydrocarbons, for example methylene chloride, chloroform, esters or ethers, for example ethyl acetate, butyl acetate, amyl acetate or In this case, in tetrahydrofuran and especially in dioxane. If the reacting amine of the formula HA is liquid, the addition of a solvent may be omitted. It is expediently used; excess (5 to 10 moles) of the amine. The reaction is preferably carried out at room temperature or, for example, slowly; at an elevated temperature, for example at a temperature between room temperature and 1-10 ° C. The progress of the reaction can be monitored by thin layer chromatography.

In the reaction of 3-1-piperazinyl-riphamycin S and / or SV or its copper C-methyl homologues with said alkylating agent, especially alkylating agents of the formula XR are used, where X is halogen, for example chlorine; bromine or iodine, or the remainder of acidic inorganic acids, for example certain sulfuric or sulfuric acids, acidic acids, in particular acids ^; · Fluorosuron. Such an alkylation agent. the reagents are, for example, haloalkyls, such as the bromides, iodides or chlorides of the hydrocarbon radical R or R-mono- or ^β- ester of sulfuric acid or fluorosulfonic acid.

The reaction of said rifamycin compound with these alkylating agents is preferably carried out in the presence of a base, in particular a strongly basic, non-nucleophilic tertiary amine, in particular an amine of the formula

Xi /

X — N \

X2 kde

X is lower alkyl and each of X 1 and X 2 is a bulky aliphatic hydrocarbon radical. The groups X 1 and X 2 are, for example, lower alkyl groups having 1 to 12 carbon atoms, preferably 1 to 7 carbon atoms having a branched hydrocarbon chain, while X preferably represents a lower alkyl group having 1 to 7 carbon atoms. In particular, the so-called Hunig's base, i.e. ethyldiisopropylamine, is used. The reaction is preferably carried out in an inert solvent such as, in particular, a chlorinated aliphatic hydrocarbon, for example methylene chloride, or an alcohol, for example methanol, at temperatures between room temperature and about 100 ° C, 1 mol of alkylating agent is used per mol of rifamycin compound. and also the base is preferably added in one molar ratio. The duration of the reaction varies according to the reactants and may range from half an hour to 24 or 48 hours.

The starting materials for the above-described processes are known or can be prepared in a manner known per se. For example, 3-piperazinorifamycin SV, useful as a possible starting material, is obtained from the method of rifamycin S and piperazine as described in the aforementioned French patent and additional reduction with ascorbic acid.

The reaction product of the reaction is often present partly in the form of quinone and partly in the form of hydroquinone. It is preferred that the reaction product be unified in this regard prior to processing, i.e. the existing hydroquinone is oxidized to quinone or the existing quinone is reduced to hydroquinone, whereupon the obtained 3-amino derivative is then isolated in one or the other form. The oxidation is preferably carried out with inorganic oxidizing agents, preferably potassium ferrocyanide, the reduction preferably with ascorbic acid or sodium dithionite.

The isolation of the reaction product from the reaction mixtures thus obtained or from the reaction mixtures obtained immediately is carried out in a manner known per se, for example by dilution with water and / or optionally neutralization with an aqueous acid, for example a mineral acid or preferably citric acid. for example a chlorinated hydrocarbon such as chloroform or methylene chloride, the reaction product being transferred to the organic phase from which it can be obtained by conventional means, i.e., drying, evaporation and crystallization and / or chromatographic or other purification processes in pure form.

The quinones or hydroquinones thus obtained can be easily converted into one another, for example by treatment with the abovementioned reducing or oxidizing agents.

Quinones are mostly violet-red colored compounds. Hydroquinones are mostly colored yellow and crystallize well. Hydroquinones form metal salts, for example alkali metal salts. With acids, quinones and hydroquinones form addition salts and optionally also quaternary ammonium salts, in particular with esters of lower alkanols with hydrohalic acids, sulfuric acids or sulfonic acids. For the preparation of acid addition salts, acids which are particularly suitable for the formation of therapeutically useful salts are used in particular. These include, for example, hydrogen halides, sulfuric acids, phosphoric acids, nitric acid, perchloric acid, aliphatic, alicyclic, aromatic or heterocyclic carboxylic or sulfonic acids such as formic, acetic, propionic, succinic, glycolic, lactic, malic acids , tartaric, citric, ascorbic, maleic, hydroxymalein or pyruvic acid, phenylacetic acid, benzoic acid, p-aminobenzoic acid, anthranilic acid, p-hydroxybenzoic acid, salicylic or p-aminosalicylic acid, ammonium, methanesulfonic, ethanesulfonic, hetesulfenesulfonic, toluenesulfonic, naphthalenesulfonic or sulfanylic acid, also methionine, tryptophan, lysine or arginine.

These or other salts of the novel compounds, such as picranes, can also serve to purify the bases obtained by converting the bases into salts, separating them and releasing the bases from the salts. Because of the close relationship between the bases in the free form and in the form of their salts, it is to be understood in the foregoing and the following that the free bases are to be understood according to the meaning and purpose of the salt, if any.

The novel compounds can be used, for example, in the form of pharmaceutical compositions. These compositions comprise said compounds in admixture with a pharmaceutical organic or inorganic, solid or liquid carrier suitable for enteral, topical or parenteral administration. Suitable carriers for the carrier are those which do not react with the novel compounds produced, such as water, gelatin, lactose, starch, stearyl alcohol19925?

14 hol, magnesium stearate, talc, vegetable oils, benzyl alcohols, gum, propylene glycol, polyalkylene glycols, vaseline, cholesterin or other known drug carriers. For example, the pharmaceutical compositions may be in the form of tablets, dragees, ointments, creams, capsules, or in liquid form as solutions, suspensions, or emulsions. Optionally, they are sterilized and / or contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solubilizing aids or salts for varying the osmotic pressure or buffering agent. These compositions may also contain other therapeutically valuable substances. The compositions are obtained by conventional methods.

The novel compounds can also be used in veterinary medicine in one of the above forms.

The invention also relates to those embodiments of the above-described process for the preparation of novel 3-aminorifamycin compounds starting from a compound obtainable at some stage as an intermediate and carrying out the missing process steps or starting materials formed under the reaction conditions.

The invention is illustrated by the following examples. Temperatures are given in degrees Celsius.

Example 1 g of rifamycin S are dissolved in a mixture of 100 ml of dioxane and 96 g of 1-isobutylpiperazine and the solution is allowed to stand for 28 hours at room temperature. After this time, water was added to the solution, the solution was acidified with citric acid, and the reaction product was taken up in chloroform. The chloroform solution was washed with brine, dried (MgSO4) and evaporated. The residue was dissolved in methanol, an aqueous concentrated ascorbic acid solution was added dropwise until light yellow in color, and the reaction mixture was allowed to stand. Shortly, 3- (4-isobutyl-1-piperazinyl) -rifamycin SV precipitates as yellow crystals which, after recrystallization twice from a mixture of chloroform, methanol and water, melted at 170 ° C. Further amounts of this compound are in both last mother liquors.

UV spectrum in 0.01 N alcoholic HCl, maximum in nm (log ε): 228 (4.53), 298 (4.28), 433 (3.90).

Example 2

Analogously to Example 1, 100 g of 1- (2-ethylbutyl) piperazine was added to 90 g of rifamycin S to give 3- [4- (2-ethylbutyl) -1-piperazinyl] -trimycin SV after completion of the reaction.

Example 3

In a similar manner to Example 1, 90 g of rifamycin S is reacted with 100 g of 1- (2-methylbutyl) piperazine to give 3- [4- (2-methylbutyl) -1-piperazinyl] rifamycin SV.

He attaches

Pharmaceutical compositions containing 3- (4-isobutyl-1-piperazinyl) -rifamycin SV as a capsule antibiotic.

Ingredients:

3- (4-isobutyl-1-piperazines) rifamycin SV 50.00mg lactose 100.00mg ethylcellulose 1.50mg stearic acid 1.50mg

153.00 mg

Éříprava:

1) The active substance is mixed with lactose.

2) Ethylcellulose is dissolved in 10 times the amount of methylene chloride.

(3) The mixture (1) is moistened with solution (2) and passed through a sieve with 3 to 5 mm apertures and then dried at a temperature not exceeding 40 ° C.

4) Pass the dry granulate through a 0.5 mm sieve and blend with stearic acid powder.

The resulting mixture is then filled with size 2 capsules.

Example 1 a d 5

To a solution of 30 g of rifamycin S with 100 ml of dioxane was added 30 g of 1-cyclohexylmethylpiperazine and the mixture was allowed to stand at room temperature until the initial violet-blue color of the reaction mixture turned orange-brown. The reaction product is taken up in chloroform. After drying and evaporating the chloroform solution, the residue is dissolved in a small amount of methanol and a concentrated aqueous solution of ascorbic acid is added dropwise until the initial dark color of the methanol solution turns golden yellow. After standing briefly, 3- (4-cyclohexylmethyl-1-piperazinyl) -rifamycin SV crystallized, which was recrystallized from chloroform / methanol / water twice after purification. M.p. 176-178 ° C. In the mother liquor after the first crystallization, in addition to the reaction product produced, rifamycin SV is almost exclusively.

UV spectrum in 0.01 N alcoholic HCl, peak in nm. (log ε): 228 (4.53), 298 (4.28), 433 (3.90).

199;

Example 1 a d '6

In a similar manner, as described in the previous example, it is formed from 25 g of riamycin-S in 40 ml of dioxane and from -30. - g - (4-cycloprophylmethyl-piperazinyl) rifamycin - - - - - - - which is obtained after three recrystallization from a mixture of chloroform, methanol and water - yellow crystals having a melting point of 217 degrees Celsius.

UV spectrum · v.- 0.01 · - Ni. Alcohol -HCl, λmax: - (log s): -225 - (4.54), 272 - (4.30), 430 (3.81).

Example 7

Pharmaceutical preparations,. - containing 3s (4-Gakcoheeylmethyl-piperazinyl) -1-rifamycin SV as an antibiotic in the form of a capsule.

.Ingredients:

3- (-4-Cycloisexylmethyl-1-piperazryl) rifamycin SV 100.00 mg lactose - 50.00 mg ethylcellulose -. 1.50 mg stearic acid 1.50 mg: 153.0 µg

Production:

1) Active - the substance is mixed with szlactose.

2) Ethylcellulose is. - dissolves - in - 10 times the amount of methylene chloride.

(3) The mixture (1) is moistened with the solution (2), then passed through a sieve having a mesh size of 3 to 5 mm and then dried at a temperature not exceeding 40 ° C.

4) The dry granulate is passed through a sieve having a mesh size of 0; 5. mm, and then mixed with powdered stearic acid. . - - The resulting - - mixture is: - then - filled - retractable. capsule size. 2.

Example 1 - 8

- g rifamycin - S ·. - dissolve in a mixture of 100 ml of dioxane and 27 g of N-methallylpiperazine and allow the resulting solution to stand for 35 hours at room temperature. The water is added to the solution and the solution is added. It is acidified with citric acid and the reaction product is taken up in chloroform. The chloroform solution is washed with a sodium chloride solution, dried over sodium sulphate and evaporated. - -The rest. it dissolves in methanol; To the - - solution - - - - - was added dropwise an aqueous - - concentrated ¹ - solution. - - - ascorbic acid, - to - the solution - light - - turns color,. and - the solution - is allowed to stand. 3- (4-βιξβ1 · Ιι 1 -. Piperazinyl) rifamycin: - SV - in the form of yellow crystals. - which - after - double recrystallization - from - - - a network of chloroform, - -methanol and - - water - - melts; at a temperature of 172-174 ° C. Other amounts of this compound are in both last mother liquors.

В! 1в

UV spectrum in .alpha., .Delta. (log ε) -229- (4.59js.298 (4-J ^ 1)), 435 (3.90).

Example 9

In a similar manner as in Example 8, - 10 g of rifamycin - S is reacted with - 12 g of 1-allylpiperazine, thereby obtaining 344-4ialiyl. 1-Piperazin-1-phenylamine SV - m.p. 174-177 ° C. .

UV - - MS - - -How-capture - - V - - ^{* 1} Example 8: --228 (4.60) - 297- (4.32) 4435 - (3.91).

Example 10

Obdobnýmz-way -jako- in -Example 10- 8 g of rifamycin ^S-1 was allowed to react - with - -10 g of N- (2-methyl-2-pentenyl) piperazine; - '· .čímž'- give 3- - [4- -1-Piperazinyl] -rifamycin- -SV, m.p. '

UV spectrum captured: as in Example 8: -228- (4,558), -298- (4.90), 435- (3.92).

Example 11

In a similar way - - as · - - in; - Example 8 - 20 g of rifamycin-S-reacted with -20 g of N- (2-ethyl-2-bulenyl) -piperazine, thereby yielding 3- ( 4- (2-ethyl-2-bulenyl) -1-piperazinyl] -rifamycin SV, m.p. 174 DEG -17 DEG-175 DEG.

UV spectrum (captured as in Example 8): - 229 - (4.60, 298 - (4.33), - ^; -435 (3.93)).

Example. 12 ......

In a similar manner, 15 g of rifamycin are dispersed in the examples, and 16 g of N- (2-ethyl-2'-h (eyenyl) are omitted. - - - - to give 3- [4- (2-ethyl-2-hexenyl) -1-piperazinyl] -1-rifamycin-SV-, m.p. 156 degrees Celsius.

UV - spectrum - captured - - - as - - - - in Example 8 :. -229- (4.61), -298- (4.33), -435- (3.94).

Example - 1 and d - 1 3

In a similar way: - as in the example-- · - 8 se. 20 g of rifamycin S is reacted with 20 g of N-2- (2,3-dimethyl-2-biphenyl) -piperazine to give 3- [4- - 2-methyl] -piperazine. (4-Dimethyl-2-butenyl) -1-piperazines-1 H-rifamycin SV, m.p. 171-141 ° C.

UV spectrum, - captured; as in Example 8: 230 (4.60), 300 (4.33), 435. (3.93).

Example 14

In a similar manner as in Example 8, 10 g of rifamycin S is reacted with 11 g of

N- (2-6 ^ 11311 ^ 1) (-piperazine, - to give - - -) is obtained

3- [4- (2-ethylbutyl) -1'-piperazinyl-1'-rifamycin

M.p. 168-170 ° C.

UV spectrum recorded as in Example 8: 228 (4.47), 299 (4.32), 435 (3.94).

Example 15

To 3 g of 3- (1-piperazinyl) -rifamycin SV is added a mixture of 55 ml of ethanol, 1 ml of water,

1.15 g of cyclobutylmethyl bromide (2 equivalents) and 2.5 ml (approximately 2 equivalents) of Hunig's base (ethyldiisopropylamine) and the solution was refluxed for 20 hours. Water was then added to the solution, the solution was acidified with citric acid, and the reaction product was taken up in chloroform. The chloroform solution was washed with brine, dried over sodium sulfate and evaporated. The residue was dissolved in methanol, a few drops of aqueous concentrated ascorbic acid solution added dropwise and the reaction mixture was allowed to stand. Shortly, 3- (4-cyclobutylmethyl-1-piperazinyl) -rifamycin SV precipitates in the form of yellow crystals which slowly decompose from a mixture of chloroform, methanol and water at a temperature above 195 degrees Celsius after recrystallization twice. Another amount of this compound is in the last two mother liquors.

UV spectrum in 0.01 N alcoholic HCl, maximum in nm (log ε): 230 (4.58), 299 (4.30), 435 (3.90).

Example 16

In a similar manner to Example 15, 5 g of 3- (1-piperazinyl) -rifamycin SV was reacted with 2.1 g (2 equivalents) of cyclopentylmethyl bromide and 2.5 ml (2 equivalents) of Hunig's base to give 3- (4). -cylklooped ^ ⁱ ^] Limi TLP ^ ^ l ^ _i-r pipe azines; L) -rifamycin SV of melting point 180-185 DEG C. (decomposition).

UV spectrum recorded as in Example 8: 228 (4.58), 298 (4.31), 435 (3.92).

Example 17

In a similar manner to Example 15, 10 g of rifamycin S was reacted with 10 g of N- (1,2-dimethylpropyl) -piperazine to give 3- [4- (1,2-dimethylthylpiperazin). 184-186 degrees Celsius -1-piperazinyl-rifamycin SV.

UV spectrum recorded as in Example 8: 230 (4.58), 299 (4.33), 435 (3.93).

Example 18

If rifamycin S is reacted as follows with the N-substituted piperazines listed in the first column of the table below, the rifamycin SV derivatives substituted at the 3-position with the corresponding Ν'-substituted 1-piperazinyl moiety, which are characterized by physical Parameters listed in the table:

Reagents

Solvent rifamylamine tin S

Reaction products temperature UV spectrum in 0.01 N melting point ° C alcoholic HCl, maximum in nm (log ε)

50 ml dioxane 25 g 35 g of N- (2-phenylpropyl) - 157 až 226 (4.59), 298 (4.32), -piperazine 158 434 (3,93) 70 ml dioxane 25 g 33 g of N- (2-methylpentyl) - 163 229 (4.59), 300 (4.32), -piperazine 437 (3,93) 70 ml dioxane 25 g 30 g of N- (cycloheptylmethyl) - 174 229 (4.59), 299 (4.31), -piperazine 432 (3.93) ' 70 ml dioxane 25 g 30 g of N- (3-methylpentyl) - 173 226 (4.58), 298 (4.30), -piperazine 433 (3,90) 70 ml dioxane 20 g 28 g of N - ((cyclohexen-1-yl)) - 170 to 228 (4.77), 297 (4.50), -methyl] -piperazine 172 435 (4.09) 100 ml 25 g 30 g of N- [2- (nobornyl) methyl] - 162 228 (4.55), 302 (4.27), dioxane -piperazine 440 (3,94) 100 ml 25 g 30 g of N- (cyclooctylmethyl) - 171 227 (4.53), 297 (4.24), dioxane -piperazine 432 (3,98) 70 ml dioxane 20 g 30 g of N- [(2-methylcyclohexyl) - 169 al -methyl] -piperazine 170 70 ml dioxane 20 g 30 g of N- (2-benzylpropyl) - 149 až 228 (4.57), 299 (4.32), -piperazine 150 435 (3,93) 30 ml dioxane 6 g 5.2 g N-neopentyl- 180 227 (4.56), 430 (3.86), piperazine 295 (Schulter) 100 ml 30 g 44 g λM3,3-dimethylbutyl] - 178 dioxane -piperazine 100 ml 30 g 40 g of N- (3-methylbutyl) - 178 až dioxane -piperazine 179 150 ml 20 g 27 g of N- (3-phenyl-2-propenyl) - 165 až 206 (4.71), 211 (4.73), dioxane of the azine 166 230 (4.63), 249 (4.60), 295 (4.32), 435 (3.93)

Production:

The amine was added to a solution of rifamycin S in dioxane and the reaction mixture was allowed to stand at room temperature until the off-blue color of the reaction mixture turned orange-brown. Water was then added dropwise and the reaction mixture was acidified with citric acid. The reaction product is taken up in chloroform. After drying and evaporating the chloroform solution, the residue is dissolved in a small amount of methanol and a concentrated aqueous solution of ascorbic acid is added dropwise until the initially dark color of the methanol solution turns golden yellow. After standing for a short time, the product obtained crystallizes out and is recrystallized from chloroform / methanol / water twice after purification. Its melting point is given in the table. In the mother liquor after the first crystallization, in addition to the reaction product produced, rifamycin SV is almost exclusively.

Substituted piperazines which serve as starting materials can be prepared generally by reacting the corresponding alkyl bromides with piperazine in a manner known per se.

In some cases it is recommended to use the appropriate tosylate instead of bromide; for example, the aforementioned N- (neopentyl) -piperazine can be obtained by reacting piperazine with neopentyl tosylate.

Example 19

730 mg of isobutyl tosylate, 930 mg of N-ethyldiisopropylamine in 11 ml of anhydrous methanol and 3.12 g of 3- (1-piperazinyl) -tramycin SV are heated under reflux for 48 hours under a nitrogen atmosphere. Then the solvent was distilled off and the residue was chromatographed on 200 g of 230 mesh silica gel. A 9: 1 mixture of chloroform and methanol was used as eluent.

Fractions containing the desired reaction product according to thin layer chromatography analysis were combined, dissolved in a mixture of 15 ml of methanol and 15 ml of chloroform, and 3 ml of 10% sodium ascorbate solution were added to the solution under nitrogen. The mixture was stirred for a further 15 minutes and then shaken with chloroform and water. The organic phase is dried over sodium sulphate, evaporated and the residue is recrystallized from a small amount of chloroform and 80% methanol. Yellow crystals of 3- (4-isobutyl-1-piperazinyl) -rifamycin SV of melting point 170 ° C are obtained.

UV spectrum in 0.01 n alcoholic HCl: maximum in nm (log ε): 228 (4.53), 298 (4.28), and 433 (3.90).

As mentioned above, the compounds produced by the process according to the invention are distinguished in particular by a very pronounced antituberculous action, as evidenced by in vivo tests on mice infected with Mycobacterium bovis Ravenel. The results of these tests are shown in the following table:

Table

Results of in vivo tests performed with the reaction products described in the previous examples on mice infected with Mycobacterime bovis Ravenel

Example reaction product ED 50 (mg / kg) in mice infected with Mycobacterium bovis Ravenel hch 1 1 ch 1 2 hch 1 3 hch 1 3 ch 1 5 hch 1 5 ch 1,2 6 hch > 2 8 hch 1 8 ch 1.3 10 hch 1.4 11 hch 1 12 hch 1.5 13 hch > 2 14 hch 1 15 Dec hch > 8 16 hch 1.1 17 hch 1 17 ch 1.3 18 compound

The reaction product of Example ED 50 (mg / kg) in mice infected with Mycobacterlum bovis Ravenel

1 hch 2 2 hch 1 3 hch 1,2 4 hch 2 5 hch 1 6 hch 1.5 7 hch <2 8 hch 1 9 hch 1.5 10 hch 2 11. 12 hch 3 13 hch 1.7

hch = ’hydroquinone form ch = quinone form

As can be seen from the results, the effective doses are very small and hence the efficacy is very pronounced. Compounds produced by the method of. The invention is one of the most potent antituberculous antibiotics and is comparable to the commercial antibiotic Rifampicin, which in many cases exceeds the intensity of its action. This approach is that they are relatively much easier to manufacture than said Rifampicin.

Claims (33)

SUBJECT A process for the preparation of novel antibiotic active compounds of rifamycin S (I) substituted at the 3-position with an aliphatic substituted amino group of the general formula I 31 31 ChK ^υ (I) and their respective hydroquinones (compound II), derivatives of rifamycin SV, which contains a naphthohydroquinone ring of nu nu instead of the naphthoquinone ring OH, wherein in the formula I I invent R is a hydrocarbon radical of formula III wherein n = 0 or 1, Z 1 represents a C 1 -C 7 alkyl radical or a C 2 -C 7 alkenyl radical which may be substituted by an unsubstituted or C 1 -C 7 alkyl radical substituted by a phenyl, cycloalkyl or cycloalkenyl group, or a phenyl radical or unsubstituted; a (C 1 -C 7) alkyl group substituted by a cycloalkyl or cycloalkenyl radical; the phenyl radicals may also be substituted with halogens, Z2 ^: represents hydrogen or an alkyl radical of 1 to 7 carbon atoms or an alkenyl radical of 2 to 7 atoms. carbon, and Z3 represents an alkyl radical of 1. and Z 2 and Z 3 together represent an alkylidene radical having 1 to 7 carbon atoms or an alkenylidene radical having 2 to 7 carbon atoms and Z 1; Z3 taken together are an alkylene or alkenylene residue, but 1 Z1-Z2 may be such an alkylene or alkenylene residue when Z3 and Z 'are a double bond, and each of Z' and Z 'is either A represents a 4R-1-piperazinyl radical, optionally substituted on carbon atoms, with C1-C7 alkyl groups in which hydrogen or a C1-C7 alkyl group, wherein in said groups optionally cycloaliphatic groups formed by an alkylene or alkenylene group Z1 + Z3 or Z1 + Z2, one or more non-adjacent C-C atomic pairs in the ring may be optionally linked directly or indirectly with an alkylidene group of 1 to 7 carbon atoms or an alkylene group, or one carbon atom of an alkylene group may be spirocyclically or R is allyl, and their salts and quaternary ammonium salts, characterized in that rifamycin S is reacted with an amine of formula HA wherein A is as defined above in an organic solvent free of hydroxyl groups, preferably he of low polarity, especially in dioxane or even without solvent, using an excess of amine, at temperatures ranging from room temperature to 100 ° C, or with 3- (1-piperazinyl) -trimycin S or SV, which may optionally be substituted with alkyl radicals of 1 to 7 carbon atoms on the piperazinyl radical, reacted with an alkylating agent suitable for the N-alkylation of the piperazines of formula XR wherein X is a halogen atom or a sulfonic acid residue or an oxygenated inorganic acid residue in the presence of a base at temperatures ranging from room temperature up to 100 ° C, using 1 mole each of the reactants in an inert solvent such as a chlorinated aliphatic hydrocarbon or a lower aliphatic alkanol, through which alkylating agent introduces the radical R, wherein R is as defined above, and the 3-amino-substitution obtained the product of rifamycin S or rifamycin SV is isolated and / or, if desired, before or after isolation, the hydroquinone obtained is oxidized to quinone or the obtained quinone is reduced to hydroquinone and / or the compound obtained is converted into a salt. 2. The process according to claim 1, wherein the base is a strongly basic, not nucleophilic tertiary amine, in the alkylation. 3. The process according to claim 2, wherein the base is ethyl diisopropylamine. 4. Process according to claim 1, characterized in that the obtained hydroquinone is oxidized to the corresponding quinone with hydrogen peroxide, ammonium persulfate or potassium ferrocyanide. 5. A process according to claim 1, wherein the quinone obtained is reduced to hydroquinone by ascorbic acid or a salt thereof. 6. Process according to claim 4, characterized in that the oxidation or reduction is carried out in the reaction mixture before the product is isolated. 7. A process according to any one of claims 1 to 6, wherein the compound containing the rifamycin moiety is reacted with 4R-1-piperazine and optionally an alkylating agent of the formula XR wherein X is halogen an oxygen inorganic acid or sulfuric acid halogen; represents one of the groups of formulas IV, V, VI or VII At ^pH ₃ (IV), (V), folk) _y P, V (VI) (VII) wherein n is 1 or 2 and each R Vi and V2 is an alkyl or alkenyl group having 1-7 carbon atoms; V3 represents hydrogen or an alkyl or alkenyl group having 1 to 7 carbon atoms, or together with V2; alkylidene or alkenylidene of 1 to 7 carbon atoms, ph represents unsubstituted or chlorine or bromine atoms and / or methyl-substituted phenyl, cy represents cycloalkyl or cycloalkenyl of 5 to 8 carbon atoms in the ring, the ring optionally being substituted by lower C 1-4 alkyl groups, py represents a cycloalkyl or cycloalkenyl group or a phenyl group such as the phenyl group as defined above, and alk represents a straight or branched alkylene group having 2 to 4 carbon atoms or an alkylidene group having 1 to 4 carbon atoms . 8. The method of claim 7, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X and R are as defined in point 7 and cycloalkyl or cycloalkenyl groups of cy or py are 5-7. 6 carbon atoms in the ring. 9. The method of claim 7, wherein 4R-1-piperazine or an alkylating agent XR is used, wherein X is as defined in point 7, and R is IV or V, wherein n is 1. 10. The process of claim 9, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is 199238 of a pin of formula IV or V wherein V3 is hydrogen. 11. A process according to any one of claims 1 to 6, wherein the compound containing the rifamycin moiety is reacted with 4R-1-piperazine and optionally an alkylating agent of formula XR wherein X is as defined in 7 and R is a cycloaliphatic-aliphatic hydrocarbon radical. . 12. The process of claim 11 wherein the cycloaliphatic moiety of R is a cycloalkyl or cycloalkenyl moiety having 3 or 5 to 8 carbon ring atoms and the aliphatic moiety is an alkyl moiety having 1 to 4 carbon atoms. 13. The method of claim 9, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is an isobutyl radical. 14. The method of claim 1, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in item 7 and R is a 2-ethyl-2-butenyl radical. 15. The method of claim 9, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is 2-ethylbutyl. 16. The process of claim 9, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is 2-methylbutyl. _F 17. The method of claim 9, wherein 4R-1-piperazine or an alkylating agent of formula XR is used wherein X is as defined in 7 and R is 2-methylpentyl. 18. The process of claim 9, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is 2-phenylpropyl. 19. The process of claim 9, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is a methallyl group. 20. The process of claim 1, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is a 2,3-dimethyl-2-butenyl radical. 21. The method of claim 11, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in item 7 and R is a cyclohexylmethyl radical. 22. The process of claim 1, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is 2-methylcyclohexylmethyl. 23. The method of claim 1, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is cyclobutyl. 24. The process of claim 1, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is cyclopentyl. 25. The process of claim 1, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is a 1,2-dimethylpropyl radical. 26. The method of claim 11, wherein 4R-1-piperazine or an alkylating agent of formula XR is used wherein X is as defined in 7 and R is a 3-cyclohexenylmethyl radical. 27. The process of claim 11, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in item 7 and R is cyclopropylmethyl. 28. The process of claim 11, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in item 7 and R is a cycloheptylmethyl radical. 29. The method of claim 1, wherein 4R-1-piperazine or an alkylating agent of formula XR is as defined in item 7 and R is 3-methylbutyl or 3-methylpentyl. 30. The method of claim 11, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in item 7 and R is a 2-cyclohexylbutyl radical. 31. The method of claim 1, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in 7 and R is cyclooctylmethyl. 32. The method of claim 1, wherein 4R-1-piperazine or an alkylating agent of formula XR is used wherein X is as defined in 7 and R is a 2-methyl-cyclohexylmethyl radical. 33. The method of claim 1, wherein 4R-1-piperazine or an alkylating agent of formula XR is used, wherein X is as defined in point 7 and R is bicyclo [2.2. 1] hept-2-yl-methyl residue.