DD140048A5 - PROCESS FOR PREPARING 4 "-EDOXY-4" -AMINO-ERYTHROMYCIN-A COMPOUNDS - Google Patents

Description

Process for the preparation of 4 ^u -sythromycin ~ A derivatives

Application of the invention:

The invention relates to a process for the preparation of 4 "-deoxy-4" amino-erythromycin A derivatives which are useful as novel antibacterial agents. The invention further relates to production processes for pharmazentrisch acceptable acid addition salts of these compounds.

Characteristic of the known technical solutions :

Erythromycin is an antibiotic obtained during the growth of a strain of Streptornyces erythreus in a suitable medium as described in US Pat. No. 2,653,899. Erythromycin, which is formed in forms A and B, has structural formula V.

The structural formula shows that the antibiotic is composed of three main parts: a sugar fragment known as cladinose, a second sugar moiety which has a basic substituent and is known as a desoamine, and a fourteen-membered lactone ring which is called erythronolide A or B or the description - as a

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crolidring is called. The numbering of the macrololid ring is made with simple numbers, those of the Desoarain with a dashed number and those of the Cladinose with two numbers.

Numerous derivatives of erythromycin have been prepared to modify its biological or pharmacodynamic properties.

In U.S. Patent 3,141? In 077, the reaction product of erythromycin and ethylene carbonate has been described as a very active antibacterial agent. U.S. Patent 3,884,903 discloses 4-deoxy-4-oxo-erythromycin A and B derivatives as useful antibiotics.

Erythromycylamine, the 9-amino derivative of erythromycin A, has been the subject of considerable research (British Patent 1,100,504, Tetrahedron Letters, 1645 (1967) and Croatica Chemica Acta, 22 " ² ? 3 (1967))" and there was some controversy in terms of its structural identity, (Tetrahedron Letters, 167 (1970) and British Patent Specification 1 y + Λ 022). SuIfonaraid derivatives of erythromycylamine are disclosed in U.S. Patent 3,983,103 as being useful as antibacterial agents. Other derivatives have also been reported (Ryden et al., J. Med. Chem., 16, 1059 (1973) and Massey et al., J. Med. Chem., 17, 105 (1974)) to have antibacterial activity in vitro and in vivo.

Explanation of the essence of the invention:

It has now been found that certain novel 4 "-deoxy-4" -araino-erythromycin A derivatives have superior properties as antibacterial agents. These compounds are represented by the formulas III and IV.

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and pharmaceutically acceptable acid addition salts thereof, wherein R. and iir are each a hydrogen atom or an alkanoyl group having up to 3 carbon atoms, R _? an alkanoyl radical having 2 to 3 carbon atoms, R-. represents a hydrogen atom, Rp and R-, when taken together, each represents the radical O.

If

-C- are and R_

and Bi, if taken together, the remainder 0 ^ * tt

-C- represent.

A preferred group of compounds within this class of chemotherapeutic agents are those of Formula III. Particularly preferred within this group are compounds in which R ₉ and R ", when taken together, are the radical

-C- represent.

A second preferred group of compounds in this class of antibacterial agents are those of formula IV. Particularly preferred within this group are compounds in which R _{1 is} a hydrogen atom and in which R "and R, when taken together, form the Rest 0

represent ti -C-.

In the second class of compounds according to the invention which are advantageous as intermediates which lead to the antibacterial agents of the formulas III and IV, the formulas I and II in which R.sup.1 is a hydrogen atom or an alkanoyl radical having 2 to 3 carbon atoms have an alkanoyl radical with 2 to 3 carbon atoms, Y is the radical

0 η

N-OH or N-O-CCH

R 1 is hydrogen and R g and R taken together are 0

Il

-C- represent.

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Preferred within this class of intermediates are the compounds of formula I. Particularly preferred within this group of intermediates are compounds wherein R. is hydrogen atom or the acetyl radical.

A second group of preferred intermediates are those of formula II. Particularly preferred within this group are the intermediates in which R.sup.2 is hydrogen, furthermore those in which R. is the acetyl radical.

Also within the scope of the invention are processes for the preparation of intermediate compounds of the formulas Ia and Ha, wherein Ac and R _{2 are} each an alkanoyl radical having 2 to 3 carbon atoms, R_ represents a hydrogen atom and R "and R_ when taken together, the radical 0

It

-C-, whereby the method thereby

characterized in that a compound of the formula I 'or II ^{1 is} reacted with 1 mol of each dimethyl sulfoxide and trifluoroacetic anhydride in a reaction-inert solvent at about -30 C to -65 C, and then the reaction mixture is brought into contact with at least 1 mol of triethylamine ,

A preferred feature of this process is the oxidation of the compounds of formula I ¹ and II 'in which the reaction-inert solvent is methylene chloride.

A second method within the invention involves the preparation of compounds of formulas Ia and Uk wherein Ac and R are each an Al

I S

kanoyl radical having 2 to 3 carbon atoms, R_ is a hydrogen atom and R ₉ and R ^ ₄ , when taken together, the radical 0

-C represent, wherein the method is characterized in that a

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Compound of formulas I ¹ or II ^{1 is} reacted with 1 mol of each N-chlorosuccinimide and dimethyl sulfide in a reaction-inert solvent at about ^{0 0} C to -25 ° C, and then the reaction mixture with at least 1 mol of triethylamine in contact. .

A preferred feature of the process of the invention is the use of toluene and enol as reaction inert solvents.

In the present invention, the stereochemical designation of the substituents on the sugars and the macrolide ring is that of the naturally occurring erythromycin A, except for the epimerization in the 4 "division where indicated.

Also within the scope of the invention are erythromycin B derivatives which correspond to those of the formula I and II. These erythromycin B compounds are useful intermediates and are prepared by the same synthetic route as described herein for the erythromycin A compounds. The erythromycin B intermediates are also converted into erythromycin B amines according to the procedures described herein, which correspond to the compounds of formulas III and IV according to the invention. The erythromycin B-amines are also useful as antibacterial agents.

The invention will be explained in more detail below.

For the methods used to synthesize the V-deoxy-α-amino erythromycin-A derived antibacterial agents of the invention, reference is made to Schemes 1 and 2, wherein of a 2α-alkanoyl erythromycin-A or a derivative is assumed.

The selective oxidation of compounds of the formula I ¹ and II ¹ . to I or

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II (Y = O) is the first of the processes according to the invention and comprises the reaction of the compounds I ¹ and II ¹ with trifluoroacetic anhydride and dimethyl sulfoxide with subsequent addition of a tertiary amine such as triethylamine.

In practice, trifluoroacetic anhydride and dimethylsulfoxide are initially combined in a reaction-inert solvent at about -65 ° C. After 10 to 15 minutes, the alcohols I ¹ and II ^{f are added} at a certain rate above -3O ⁰ C increases. At temperatures above -30 C, the complex of trifluoroacetic anhydride diraethylsulfoxide is not stable. The reaction temperature is maintained between -30 ° C and -65 ° C for about 15 minutes, then it is lowered to about -? 0 ° C. A tertiary amine is added all at once, and the reaction mixture is allowed to warm for a period of 10 to 15 minutes. The reaction mixture is then treated with water and worked up.

With respect to the amounts of reactants, 1 mole of the trifluoroacetic anhydride and the dimethyl sulfoxide are required for each mole of the alcohol substrate used. Experimentally, it is advantageous to use a 1 to 5 fold excess of the anhydride and the dimethyl sulfoxide to accelerate the completion of the reaction. The tertiary amine used should correspond to the molar amount of trifluoroacetic anhydride used.

The reaction-inert solvent used in this process is that which solubilizes the reactants appreciably and does not react with any of the reactants or the products formed to any great extent. Since this oxidation process is carried out at -30 C to ⁰ -65 ° C, it is preferred that the solvent in addition to the aforementioned properties an overall

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has a freezing point below the reaction temperature. Such solvents or mixtures thereof which meet these criteria are toluene, methylene chloride, ethyl acetate, chloroform or tetrahydrofuran. Solvents which meet the above requirements, but have a freezing point above the reaction temperature, can be used in minor amounts in combination with one of the preferred solvents. The most preferred solvent for the process is methylene chloride.

The preferred compounds prepared by this process are S ^ acetyl-1-deoxy-1-oxo-erythroraycih-A, 11,2'-diacetyl-4'-deoxy-4'-oxo-erythromycin A-6 , 9-hemiketal and 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin AS ^ -hemiketal 11,12-carbonate ester.

The reaction time is not critical and depends on the reaction temperature and the given reactivity of the starting reagents. At temperatures of about -30 C to -65 C, the reaction is completed in 15 to 30 minutes.

With regard to the order of addition of the reagents, it is preferred that the trifluoroacetic anhydride be combined with the dimethylsulfoxide, followed by the addition of the required alcohol substrate. Furthermore, it is advantageous, as already mentioned, if the temperature of the reaction is kept below -30 ° C. This is in accordance with the specifications of Omura et al., J. Org. Chem., 41, 957 (1976).

The second method of the invention, which is used to prepare intermediates useful in antibacterial agents, is represented by Scheme 3.

The second experienced represents an oxidation reaction in which

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4 ⁿ -hydroxy substituent of I ¹ and II · in which Ac and R "are each alkanoyl radicals having from 2 to 3 carbon atoms, R - represents a hydrogen atom and R ₂ and E-, when taken together, is the radical 0

-C-, is oxidized to a ^ '- deoxy - ^' - oxo-erythromycin A compounds.

The process involves the use of N-chlorosuccinimide and dimethyl sulfide as the oxidizing agent. In practice, these two reagents are first combined together in a reaction-inert solvent at about 0 ° C. After 10 to 20 minutes, the temperature is lowered to ⁰ ° C to -25 ° C, and the alcohol substrate I 'or II ¹ is added while maintaining the above-mentioned temperature; After a reaction time of 2 to 4 hours, a tertiary amine such as triethylamine is added, the reaction mixture is hydrolyzed and worked up.

With respect to the amounts of reactants, for each mole of the alcohol substrate used, 1 mole each of the N-chlorosuccinimide and the dimethylsulfide is required. Experimentally, it is advantageous to use a 1 to 20 fold excess of the succinimide and sulfide reactants to accelerate the completion of the reaction. The tertiary amine used should correspond to the molar amount of succinimide used.

The reaction-inert solvent used in this process should be a solvent which solubilizes the reactants appreciably and does not react to any appreciable degree with any of the reactants or the products formed. Since the reaction is carried out at about ⁰ ° C to -25 ° C, it is preferred that the solvent has a freezing point below the reaction temperature in addition to the aforementioned properties.

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Such solvents or mixtures thereof which meet these criteria are toluene, ethyl acetate, chloroform, methylene chloride or tetrahydrofuran. Solvents that meet these requirements but have a freezing point above the reaction temperature may also be used in minor amounts in combination with one or more of the preferred solvents. The most preferred solvent for this process is toluene-benzene.

The preferred compounds prepared by this method are: 11,2'-diacetyl-4 "-deoxy-4" -oxo-erythromycin A-6,9-hemiketal, ^2f -acetyl 1-4 "-deoxy" - oxo-erythromycin-A-6,9-hemiketal-11,12-catenate ester and 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-A.

The reaction time is not critical and depends on the concentration, the reaction temperature and the reactivity inherent in the reagents. At a reaction temperature reaction time about 2 to 4 hours.

from. At a reaction temperature of 0 C to -25 C is the

With regard to the order of addition, it is preferable, as mentioned above, when the alcohol substrate I ¹ or II ^{1 is added} to the previously prepared mixture of the succinimide derivative and the methylsulfide id.

Both of the methods described herein are considered to be unique because of the selectivity of the oxidation occurring exclusively at the 4 "hydroxy substituents, leaving the other secondary alcohol functions unaffected in the molecule.

The useful intermediates in the form of the 4 "-deoxy-4" -oxo compounds of formula II wherein R 1 and R _{2 are} each alkanoyl radicals having 2 to 3 carbon atoms and R ₁ is a hydrogenatora are prepared by treatment of a compound of formula I ₁ where Y = O and R. a

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Alkanoyl radical having 2 to 3 carbon atoms, prepared with an alkanoic anhydride (R ^ O) and pyridine.

In practice, the ketone of formula I is contacted with an excess of the anhydride in pyridine as the solvent. Preferably, an amount as large as a 4-fold excess of the anhydride is used in the reaction.

The reaction is suitably carried out at ambient temperatures. At these reaction temperatures, the reaction time etva is 12 to 24 hours.

The removal of the alkanoyl residue at the 2'-position of the intermediate ketones I (Y = O) and II is carried out by a solvolysis reaction in which the compound related to 2'-alkanoyl-4'-deoxy-4'-oxo-erythromycin A is is stirred with an excess of methanol overnight at cinnamon © rtemperatur. Removal of the methanol and subsequent purification of residual product, where necessary, provides compounds of formula I (Y s 0) and II in which R _x . represents a hydrogen atom.

As previously described, the ketones of formula I (Y = O) and II are useful intermediates which result in the 4 "-deoxy-4" -amino erythromycin A antibacterial agents of formula III and IV of the present invention. Preferred intermediates in this group are 2'-acetyl-4 ^ll -deoxy-4 "-oxo-erythromycin-A-6,9-heraiketal ^/ l1, i2-carbonate ester and 4" -deoxy-4 "-oxo erythromycin A-6,9-hemiketal 11,12-carbonate ester.

Several synthetic routes can be used in the preparation of the antibacterial agents of formulas III and IV from the required ketones I (Y = 0) and II.

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The preparation of the 4-deoxy-4-amino-erythroraycon-A compounds of formula III is carried out by condensation of the ketones II with the ammonium salt of a lower alkanecarboxylic acid and subsequent reduction of the imine formed in situ. The term "nidere alkanecarboxylic acid" used herein refers to acids having 2 to 4 carbon atoms.

In practice, a solution of ketone II in a lower alkanol such as methanol or isopropanol is treated with the ammonium salt of a lower alkanecarboxylic acid such as acetic acid, and the cooled Reaktionsgeraisch is treated with the reducing agent sodium cyanoborohydride. The reaction is allowed to proceed at room temperature for several hours before hydrolyzing the reaction mixture and isolating the product.

Although 1 mole of ammonium alkanoate per mole of ketone is required, it is preferred to use an excess as large as a 10-fold excess to ensure complete and rapid formation of the imine. Such surplus quantities appear to have little adverse effect on the quality of the product.

With respect to the amount of reducing agent to be used per mole of ketone, it is preferred that about 2 moles of sodium cyanoborohydride be used per mole of ketone.

The reaction time varies with the concentration, the reaction temperature and the reactivity inherent in the reagents. At room temperature, the preferred reaction temperature, the reaction is essentially complete after 2 to 3 hours.

When the solvent in the form of the lower alkanol is methanol, as mentioned above, a substantial solvolysis at any alcohol level occurs.

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kanoylrest in the 2 ^f- position on. To avoid the removal of such a best, the use of isopropanol as a reaction solvent is preferred.

The preferred ammonium alkanoate or atnmonium alkanecarboxylic acid salt for this reaction is ammonium acetate, as indicated above.

In isolating the desired ^ '' - deoxy - '' - amino erythromycin A derivatives from any nonbasic by-products or any starting material, the basic nature of the final product is advantageously exploited. Accordingly, an aqueous solution. of the product is extracted over a range of gradually increasing pH so that neutral or non-basic materials are extracted at lower pH values and the product at a pH greater than 5. The extraction solvents, either ethyl acetate or diethyl ether, are backwashed with brine and water, dried over sodium sulfate, and the product is obtained by removal of the solvent. Additional purification may be carried out, if necessary, by column chromatography on silica gel according to the known procedures.

As described above, the solvolysis of the 2'-alkanoyl residue from the corresponding 2'-alkanoyl-4 "-deoxy-4" -araino-erythromycin A derivative can be achieved by allowing a methanol solution of this compound to stand overnight at ambient temperature ,

During the reducing amination of ketones of the formula II, in which R ₉ and R, when taken together, form the ^K est 0

-C- represent

and R is alkanoyl of 2 to 3 carbon atoms or hydrogen, it should be noted that amines related to both compounds of formula III and IV are formed. This is done by the scheme

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4 reproduced.

The amine products III and IV as shown are conveniently separated by selective crystallization from diethyl ether. Ucrystallization of the mixture of III and IV, as shown, from acetone-water induces hemicetalation into the amine of formula IV, resulting in the isolation of III as the sole product.

The first direct route to the amine compounds of formula IV is the same as previously described and involves condensation of ketone I with an ammonium alkanoate, followed by reduction of in situ formed imine with sodium cyanoborohydride.

Compounds of Formula IV in which R., R, and R have the previously indicated definition are also prepared by reduction of the aforementioned Itnins using hydrogen and a suitable hydrogenation catalyst. Experimentally, the appropriate ketone (I) in a lower alkanol such as methanol or isopropanol is treated with the ammonium salt of a lower alkanoic acid such as acetic acid and, after addition of the hydrogenation catalyst, the mixture is shaken in a hydrogen atmosphere until the reaction is essentially complete. ,

Although 1 mole of the ammonium alkanoate is required per mole of ketone, preferably an excess of up to 10 times _{5 is} employed to ensure complete and rapid formation of the imine. Such excess levels appear to have little adverse effect on the quality of the product to own",

^The hydrogenation catalyst can be selected from a wide variety of agents; Raney nickel and 5 % palladium on charcoal charcoal), however, are the preferred catalysts. You can

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Different amounts are used, which depend on the desired speed to complete the reaction. Amounts of 10-200 % of the weight of Compound I can be effectively used.

The hydrogen gas pressure in the hydrogenation vessel also affects the reaction rate. Preferably, and because of suitable reaction times, an initial pressure of 30 atm should be used. Furthermore, it is preferred for reasons of convenience that the reduction be carried out at ambient temperatures.

The reaction time depends on a number of factors including temperature, pressure, the concentration of reactants and the reactivity inherent in the reagents. Under the preferred conditions mentioned above, the reaction is completed in 12 to 24 hours.

The product is isolated by filtering off the spent catalyst and removing the solvent in vacuo. The remaining material is then treated with water and the product is isolated from the non-basic materials by extraction of the basic product from water at varying pH's, as previously described.

As previously described, when the solvent in the form of the lower alkanol is methanol, substantial solvolysis of any alkanoyl group occurs at the 2'-position. To avoid removal of such residue, the use of isopropanol as a reaction solvent. prefers,

, the second synthesis route to the ^ '- deoxy - ^ - amino erythromycin A antibacterial agents of formula IV comprises the initial conversion

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ketones of formula I (Y = O) into an oxime or oxime derivative, i.

rf

Y s N-OH or N-O-CCH ", followed by the reduction of the Oxitns or its derivative followed.

The oximes of ketones I (Y = 0) are prepared by reacting these ketones with hydroxylamine hydrochloride and β-carbonate in methanol or isopropanol at room temperature. In practice, the use of an excess of hydroxylamine is preferred, and as much excess as a 3-fold excess gives the desired intermediate in good yields. The use of ambient temperatures and an excess of the hydroxylamine allows the preparation of the desired oxime derivative in a reaction period of 1 to 3 hours. The barium carbonate is used in 2 times the molar amount as the hydroxylamine hydrochloride used. The product is isolated by addition of the reaction mixture to water and subsequent basification to pH = 9.5 and extraction with a water immiscible solvent such as ethyl acetate.

Alternatively, the reaction mixture can be filtered and the filtrate evaporated to dryness in vacuo. The residue is then partitioned between water at pH = 3.0 - 9.5, and distributed with a V / ater-immiscible solvent.

The preparation of the O-acetyloxime compounds of the formula I (Y = NO-CCH.) Is carried out by acetylation of the corresponding oxime. Experimentally, 1 mole of the oxirane is reacted with 1 mole of acetic anhydride in the presence of Ί mole of pyridine or triethylamine. The use of an excess of the anhydride and pyridine facilitates completion of the reaction and an excess of 30-40 % is preferred. The reaction is best carried out in an aprotic solvent such as benzene or ethyl acetate at room temperature overnight. At the completion of the reaction, water is added, the pH becomes 9.0

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adjusted, and the product is separated in the solvent layer. The preferred oximes and oxime derivatives, which are useful intermediates resulting in the antibacterial agents derived from 4- "~ deoxy-A" -amino-erythromycin A, include: 2'-acetyl-1-deoxy-3-oxo erythromycin A oxime, 2'-A ce ty 1-4 "-de oxy-4 ^{M-oxo-erythromycin-A-0} ~ acetyloxira, 4 ^ll -deoxy-4" ~ oxo-erythromycin A oxime and 4 "deoxy ~ 4" -oxo-erythroraycin-A-0 acetyloxime.

If

The reduction of the ketone derivatives (Y = N-OH or NO-CCH-) is carried out by catalytic hydrogenation, in which a solution of the oxime or derivative thereof in a lower alkanol such as isopropanol and © in Raney nickel catalyst in a hydrogen atmosphere at a Initial pressure of 70.3 atm at room temperature above N eight. Filtration of the spent catalyst followed by removal of the solvent from the filtrate gives the isolation of the desired 4-deoxy-4-amino-antibacterial agent according to formula IV. If methanol is used as solvent in this reduction, solvolysis is used a 2'-alkanoyl probably. To avoid this side reaction, isopropanol is used.

Preferred among these 4 "-deoxy-4" -amino-erythromycin-A derived antibacterial agents of the formula III and IV are both epimers of the 4 "-deoxy-4" -amino-erythromycin A-6 _l 9-hemiketal. " 11 _{L of} 12-carbonate ester and 4 "-deoxy-4" -amino-erythromycin A and of the 4 "-deoxy-V ^T -araino-erythroraycin- ^ A-1'1, i2-carbonate ester.

Of course, in utilizing the chemotherapeutic activity of such compounds of formula III and IV according to the invention which form salts, the use of pharmaceutically acceptable salts is preferred. Although the water solubility, high toxicity or lack of crystalline properties is a special type of salt

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may be unsuitable for use as such in a given pharmaceutical application, the water-insoluble or toxic salts may be converted to the corresponding pharmaceutically-acceptable bases by decomposition of the salt as described above, or alternatively they may be incorporated in US Pat Any desired pharmaceutically acceptable acid addition salt may be converted.

Examples of acids which provide pharmaceutically acceptable anions are: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid or sulphurous acid, phosphoric acid, acetic acid, lactic acid, citric acid, tartaric acid, bersetinic acid, maleic acid, gluconic acid and aspartic acid.

As described above, the stereochemistry of the starting materials resulting in the antibacterial agents of the invention corresponds to that of the natural material. The oxidation of A - "- hydroxyl group to a ketone and subsequent conversion of this ketone to the V-amines will stall a possibility for change of the stereochemistry of the 4 ⁿ -Substitueηteη from that of the natural product is away Therefore, it is when the compounds I. (Y = O) and II are converted to amines by any of the methods described above, it is possible to form two epimeric amines Experimentally, it has been found that both epimeric amines are present in the final product in varying proportions, depending on the choice of synthesis step. If the isolated product consists predominantly of one of the epimers, this epimer can be purified by repeated recrystallization from a suitable solvent to a constant melting point The other epimer, namely the lesser amount of the originally isolated solid material, is the epimer overwhelming product in the mother liquor.It may be from this obtained by methods known per se, for example by evaporating the mother liquor and

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repeated recrystallization of the residue to a product of constant melting point.

Although this mixture of epimers can be separated by methods known per se, for practical reasons it is advantageous to use this mixture as it is isolated after the reaction. However, it is often advantageous to purify the mixture of epimers by at least one recrystallization from a suitable solvent, subjecting it to high pressure liquid column chromatography, solvent distribution or trituration in a suitable solvent. This purification, while not necessarily separating the epimers, removes such foreign materials as raw materials and unwanted by-products.

The absolute stereochemical assignment of epimers has not yet been completed. However, both epimers of a given compound show the same type of activity, e.g. as antibacterial agents.

The novel ^ "-deoxy-V-amino-erythrornycin A derivatives described herein exhibit in vitro activity against a variety of Gram-positive microorganisms, e.g. Staphylococcus aureus and Streptococcus pyogenes and against certain Gram-negative microorganisms such as those of spherical or ellipsoidal shape (cocci). Their activity is readily demonstrated by in vitro assays against various microorganisms in a brain heart infusion medium following the usual double serial dilution procedure. The in vitro activity makes them available for topical application in the form of ointments, creams and the like, for sterilization purposes, e.g. B, objects in disease conditions, and as industrial, antimicrobial agents, e.g. suitable for the treatment of water, the control of sludge, the preservation of paints and wood,.

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For use in vitro, e.g. for a topical application, it is often advantageous to mix the selected product with a pharmaceutically acceptable carrier and / or diluent such as a vegetable or mineral oil or a moisturizing cream. Equally, they may be dissolved or dispersed in liquid carriers or solvents, e.g. in water, alcohol, glycols or mixtures thereof or in other pharmaceutically acceptable, inert media, i. Media that have no harmful effect on the active ingredient. For such purposes, it is generally acceptable to use concentrations of active ingredients of from about 0.01% to 10% by weight, based on the total composition.

Furthermore, many of the compounds of the invention and their acid addition salts are active against Gram-positive and certain Gram-negative microorganisms, e.g. Pasteurella multocida and Neisseria sicca, in vivo via oral and / or parental routes of administration in animals and in humans. Their in vivo activity is more limited with respect to susceptible organisms and determined by the usual procedure which involves the infection of mice of practically equal weight with the test organism and their oral or subcutaneous subsequent treatment with the test compound. In practice, in mice, e.g. 10 animals were subjected to an intraperitoneal vaccination of appropriately diluted cultures containing approximately 1 to 10 times the LD ". (The lowest concentration of organisms required to cause 100/5 deaths). Control tests are performed concurrently in which the mice are vaccinated with lower dilutions to test for a possible variation in organism virulence. The test compound is applied 0.5 hours after seeding and repeated 4, 24 and 48 hours later. Dia surviving mice are kept for 4 days after the last treatment, and the

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Number of surviving animals is determined.

When used in vivo, the novel compounds of the present invention can be administered orally or parenterally, e.g. by subcutaneous or intramuscular injection, at a dosage of about 1 mg / kg to about 200 mg / kg of body weight per day. The preferred dose range is about 5 mg / kg to about 100 mg / kg body weight per day, and a particularly preferred range is from about 5 mg / kg to about 50 rag / kg body weight per day.

Carriers suitable for parenteral injection may be either aqueous carriers such as water, isotonic saline, isotonic dextrose solution, Ringer's solution, or non-aqueous vehicles such as vegetable oils (cottonseed oil, peanut oil, corn oil, sesame oil), dimethylsulfoxide or other non-aqueous vehicles containing the therapeutic agents Effectiveness of the preparation and are non-toxic in the applied volume and the applied proportion (glycerol, propylene glycol, sorbitol), be. In addition, suitable compositions for the unprepared preparation of solutions prior to application can be advantageously prepared. Such compositions may include liquid diluents, e.g. Propylene glycol, diethyl carbonate, glycerine, sorbitol, etc., buffering agents, hyaluronidase, local anesthetics, and inorganic salts to provide the desired pharmacological properties. The compounds may also be combined with various pharmaceutically acceptable inert carriers including solid diluents, aqueous carriers, non-toxic organic solvents in the form of capsules, tablets, lozenges, troches, dry mixes, suspensions, solutions, elixirs, and parenteral solutions or suspensions. In general, the compounds in different. Dosage forms are applied at concentration levels ranging from about 0.5% to about 90% by weight of the total composition.

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Embodiments:

The invention will be explained in more detail with reference to the following examples.

example 1

2'-acetyl-4-deoxy-3-oxo-er-thromycin-A

To 3 ml of methylene chloride and 0.328 ml of dimethylsulfoxide, which were cooled to about -65 ° C and kept under a nitrogen atmosphere, are added, 0.652 ml of trifluoroacetic anhydride. After about

A white slurry forms for 1 minute, indicating the presence of the complex of trifluoroacetic anhydride diraethyl sulfoxide. To the resulting slurry is added dropwise a solution of 1.0 g

2'-Acetyl-erythromycin-A-ethyl acetate, obtained by recrystallization of 2-acetyl-erythromycin-A from ethyl acetate, in 7 ml of methylene chloride, keeping the temperature at about -65 ° C. The resulting mixture is stirred for 15 minutes at about -60 C, then it is cooled to -30C. 1.6 ml of triethylamine are added quickly to the reaction mixture and the cooling bath is removed. After 15 minutes of stirring, the solution is added to 10 ml of water and the pH of the aqueous phase is adjusted to 10. The organic phase is separated off, washed successively with water (3 × 10 ml) and brine (1 × 10 ml) and dried over sodium sulfate. Removal of the solvent under reduced pressure gives 929 mg of the crude product. Recrystallization from methylene chloride-hexane gives 320 r of the purified product; F. 1O5-1O8 ° C,

(/, CDCl _3): 3.28 (3H) s, 2.21 (6H) s and 2.03 (3H) s..

Similarly, using 2'-propionylerythromycin A-ethyl acetate and following the procedure described above, 2-propyl-4-deoxy-4-oxo-erythromycin-A is obtained.

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Example ^ 2 4 ^l 'DeoX2jr-4 ^tl -oxo-erythromycin-A

A solution of 4.0 g of 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-A in 75 ml of methanol is stirred at ambient temperature for 20 hours. The solvent is removed in vacuo and the residual white foam is recrystallized from methylene chloride-hexane to give 3.44 g of product of mp 170.5-172.5 ° C.

NMR (<T, _CDCl3): 3,3δ (3H) s and 2.33 (6H) s. A product identical to this product is isolated when 2 "-propionyl-4" -deoxy-4 "-oxo-erythromycin-A is treated with methanol at room temperature.

Example_3

2 ^l -Acety_l ~ 4 "-deoxy_-4" -oxo-ery_thromy_cin-A

To a stirred solution of 13 _f 7 g of 4 "-deoxy-4" -oxo-erythromycin-A in 100 ml of ethyl acetate is added 2.3 ml of acetic anhydride, and the resulting reaction mixture is stirred at room temperature for 2 hours. The solution is added to 100 ml of water and the pH of the aqueous phase is raised. 9.5 raised by the addition of 6N sodium hydroxide solution. The organic layer is separated, dried over sodium sulfate and concentrated to give 14.5 g of a white foam which, after recrystallization from methylene chloride-hexane, gives a product identical to the product of Example 1.

Beispiel_4

2 * "Acetyl-4 ^lt -deoxy" 4 "-oxo-erythromycin A oxime

To 5OO ml of methanol, 10.8 g of 2 ^l-acetyl ~ 4-deoxy-4 ^{M ll-oxo-erythromycin-A,} 1.94 g of hydroxylamine hydrochloride and 11.0 g Bariümcarbonat

203 547

is added, and the resulting suspension is stirred at room temperature for 3 5 5 hours. The mixture is filtered and the filtrate is concentrated under reduced pressure. The remaining foam is taken up in ethyl acetate, which is then washed with water at pH = 9i5. The organic phase is separated, dried over sodium sulfate and concentrated in vacuo to give 10.6 g of the desired product.

NMR (<T \ CDCl): 3.33 (3H) s, 2.30 (6H) s and 2.06 (3H) s.

Example 5

2'-Acetyl-^ -deoxy-A ^ -oxo-erythromycin-A-O-acetyloxime

To a solution of 330 mg of 2 ^l -acetyl-4 "-deoxy-4" -oxo-erythrotnycin ~ A-oxime in 30 ml of ethyl acetate is added with stirring 64.2 μl of saccharic anhydride and the reaction mixture is stirred overnight at room temperature. An additional amount of 15.8 μl of acetic anhydride and 23.4 μl of triethylamine are added and stirring is continued for 4 hours. The reaction mixture is added to water and the pH is adjusted to about 9 °. The ethyl acetate layer is separated, dried over sodium sulfate and concentrated in vacuo to give 300 r of the desired product.

NMR ((T, CDCl _3): 3.38 (3H) s, 2.25 (6H) s 2.20 (3H,) s, 2, O5 (3H) s.

and 1.56 (3H) s.

In the same V / else be when using S'-propionyl-V'-deoxy-4 "-oxo-erythromycin A 4 oxira and" ^{-deoxy-4-oxo-tl} erythroiaycin ~ A ~ oxime. instead of the 2 '~ acetyl-4 ^lf -deoxy-4 "-oxo-erythromycin A oxime in the procedure described above, the corresponding O-acetyl derivatives prepared.

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example

2fAcetyl ^H-4 -deoxy-4 "-amino-erythromycin A

A mixture of 14.0 g of 2 ¹ -acetyl-4 ^ll deoxy-4 "-oxo-erythromycin A ~ 0-acetyloxime and 60 g of isopropanol-washed Raney nickel in 400 ml of isopropanol is placed in a hydrogen atmosphere at an initial pressure. The catalyst is filtered off and the filtrate is concentrated to a white powder, the residue is redissolved in 400 ml of idopropanol and washed with 50 g of fresh, isopropanol-washed Ranay nickel Hydrogenation is continued overnight at room temperature and an initial hydrogen pressure of 70.3 ata The catalyst is filtered off and the filtrate is concentrated to dryness in vacuo to give 8.1 g of the desired product.

Example ^?

Using the corresponding O-acetoxy-oxime as the starting material and following the procedure of Example 6, the α, β-aminom erythromycin A-analogous derivatives of Formula VII were prepared.

example

4 "-Deoxy_-4" -araino-erythromycin A

A solution of 2.17 g of ^2f -acetyl-4 "-deoxy-4" -araino-erythromycin-A in 50 ml of methanol is stirred at room temperature overnight. The solvent is removed under reduced pressure and the residual foam is treated with a mixture of 50 ml of chloroform and 50 ml of water. The pH of the aqueous layer is adjusted to 9.5, and the organic layer is separated. The chloroform layer is treated with fresh water and the pH is raised

-25-203 547

4.0 set. The pH of the acidic aqueous layer containing the product is adjusted gradually to 5 »6, 7» 8 and 9 by addition of base, extracting with fresh chloroform at each pH. Extracts at pH = 6 and 7 contain the major portion of the product, and these extracts are combined and treated with fresh water at pH = 4. The aqueous layer is again adjusted to pH's 5, 6 and 7, extracting with fresh chloroform at each pH. The chloroform extract at pH = 6 is dried over sodium sulfate and concentrated to give 249 mg of the product as a spiro mixture.

NMR (π, CDCl J: 3.30 (iH) s, 3.26 (2H) s, 2.30 (6H) s, and 1.46 (3H) s.

Similarly, "-deoxy-4" -amino-erythromycin A is prepared by Methanolsolvolyse of 2 ^f propionyl ~ 4 "-deoxy-4" ~ araino-erythromycin-A. 4

example ^

4 "-Deöxy 4-amino ~ ⁿ ~ erythromycin A

To a stirred solution of 3.0 g of 4 "-deoxy-4" oxo-erythromycin-A in 30 ml of methanol under a nitrogen atmosphere is added 3 »16 g of dried ammonium acetate, and after 5 min 188 mg of sodium cyanoborohydride are added to the reaction rinsed with the aid of 5 ml of methanol, and the reaction mixture is stirred at room temperature overnight. The light yellow solution is poured into 3OO ml of water and the pH is adjusted to 6.0. The aqueous layer is at pH = 6; 7; 7.5; 8th; 9 and 10 extracted using 125 ml of diethyl ether with each extraction. The extracts at pH 8, 9 and 10 are combined and washed with 125 ml of fresh water. The separated, aqueous layer is washed with ether (1 × 100 ml) at pH = 7.5, ethyl acetate (1 × 100 ml) bai pH = 7,

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Ether (1 × 100 ml) at pH = 7.5, ethyl acetate (1 × 100 ml) at pH = 7.5 and ethyl acetate (1 × 100 ml) at pH = 8, 9 and 10 extracted. The ethyl acetate extracts at pH = 9 and 10 are combined together, washed with a saturated saline solution and dried over sodium sulfate. Removal of the solvent in vacuo gives JO mg of an epimeric mixture of the desired product as off-white foam,

Example 10

V-Deoxy_-4 "amino-ery_thromycin-A (single epimeres)

A solution of 10.0 g of the epimeric mixture of 2'-acetyl-4 · "oxy-4" -amino-erythromycin-A in 150 ml of methanol is stirred at room temperature under nitrogen for 72 hours. The solvent is removed in vacuo and the residue is dissolved in a stirred mixture of 150 ml of water and 200 ml of chloroform. The aqueous layer is discarded and 150 ml of fresh water are added. The pH of the aqueous layer is adjusted to 5, and the chloroform layer is separated. The pH of the aqueous phase is adjusted sequentially to 5 »5; 6; 7; .8 and 9 are adjusted, after each adjustment with 100 ml of fresh chloroform is extracted. The chloroform extracts at pH = 6; 7 and 8 are combined together, washed successively with water and a saturated saline solution and dried over sodium sulfate. Removal of the solvent under reduced pressure gives 2.9 g of an epimer mixture of 4 "-deoxy-4" -amino-erythromycin-A. A sample of 1.9 g of the mixture is triturated with diethyl ether, with a small amount of the undissolved foam being made to crystallize. The solids are filtered off and dried to give 67 mg of a single epimer of 4 - "- deoxy-4" -amino-erythromycin-A with F, 140-14-7 C.

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Beispie1_11

1 Ί _λ 2 · diacetyl-Λ "-deoxy-4" -oxo-er ^ throraycin-A-6,9-hemiketal

Sine solution of 10 g 2 ^f-acetyl-4 "-deoxy-4" -oxo-erythroraycin-A in 250 ml of pyridine is treated with 40 ml of acetic anhydride, and the reaction mixture is allowed to stand at room temperature for 10 days. The bulk of the solvent is removed in vacuo and the remaining concentrate is added to a mixture of 150 ml of water and 100 ml of chloroform. The pH of the aqueous layer is raised to 9.0 and the chloroform is separated, dried over sodium sulfate and concentrated to dryness.

NMR (cT, CDCl _3): 3.33 (3H) s, 2.26 (6H) s, 2.10 (3H) s, 2.03 (3H) s and 1.55 (3H) s.

Example 12

Using the corresponding 4 ^ll -deoxy-4 "-oxo-erythrotnycin-A as the starting material and the corresponding alkanecarboxylic anhydride and using the procedure of Example 11, the compounds of formula VIII were synthesized.

Beispiel__i3

11-acetyl-A "-deoxy-4 ^M -oxo-erythroiylin-A ~ 6,9-he; ni-ketal

A solution of 3 »g of 11,2'-diacetyl-^ -deoxy-JV-oxo-erythromycin A-6,9-hemiketal in 50 ml of methanol is stirred under a nitrogen atmosphere overnight. The solvent is removed in vacuo to give 3 g of the desired product in the form of a yellow foam.

NMR (cT, CDCl _3): 3.35 (3H) s, 2.31 (6H) s, 2.13 (3H), and i, 55 (3H) s

-28- 203 547

Similarly, the compounds of Example 12 are prepared according to the procedure of Example 13 in 11-acetyl-4 "-deoxy-4" -oxo-erythromycin A-6 _t 9-hemiketal and 11-propionyl-4 "deoxy-4 "oxo-erythromycin A-6 _t 9-hemiketal converted.

Beisgiel_i4 11-acetyl-4 ^l '-deoxy-4 "-amino-erythromycin _A-6-i 9-hemiketal

To a stirred solution of 4.4 g of 11α-acetyl-4 "-deoxy-4" -oxo-erythromacin ~ A-6,9-hemiketal and 4.38 g of ammonium acetate in 75 ml of methanol is added 305 mg of 85% Added sodium cyanoborohydride. After stirring overnight at room temperature, the reaction mixture is poured into 3OO ml of water, to which then 250 ml of chloroform are added. The pH of the aqueous layer is adjusted to 9.8, and the chloroform layer is separated. The aqueous layer is re-extracted with chloroform and the chloroform extracts combined, dried over sodium sulfate and concentrated to a white foam. The residual foam is dissolved in a stirred mixture of 125 ml of water and 125 ml of fresh chloroform, and the pH is adjusted to 4.9. The chloroform is separated and discarded, and the aqueous layer is adjusted to pH = 5; 6; 7 and 8 is set, which is extracted after each adjustment with fresh chloroform. The extracts from the aqueous layer at pH = β and 7 are combined together, washed with a saturated saline solution and dried over sodium sulfate. Removal of the solvent gives 1.72 g of the desired product as a white foam. The product is dissolved in a minimum amount of diethyl ether and then treated with hexane until cloudy. The crystalline product which forms is filtered off and dried to give 1.33 g of product of mp 204.5-206.5 ° C. NMR C </>, CDCl3): 3.31 (2H) s, 3.28 (iH) s, 2.31 (6H) s, 2.11 (3H) s and

1.5 (3H) s.

- ² ^ - 203 547

Example 15

The procedure of Example 14 is repeated, starting from the corresponding 4 "-deoxy-4" -oxo-erythromycin-A and instead of methanol, isopropanol is used as reaction solvent to obtain the compounds of formula IX.

Example 16

ZJ-acetyl-erythromycin A-O-hethnicetal-H, 12-carbonate ester

To a solution of 13 g of erythromycin A-δ-hemiketal H. 1-carbonate ester (US Pat. No. 3 4-17077) in 150 ml of benzene is added 1.8 ml of acetic anhydride and the reaction mixture is dissolved in 200 ml 1 ml of water and the aqueous phase is basified to pH = 9.0. The benzene layer is separated off, dried over sodium sulfate and concentrated in vacuo to give 15 g of a white foam. Trituration with 50 ml of diethyl ether of the foam. Filtration and drying of the product gives 12 g of pure product F. ο 224-, 5-228.5 ⁰ C.

NMR ( _c r, CpCl_): 3.3 $ (3H) s, 2.30 (6H) s, 2.06 (3H) s and 1.61 (3H) s.

Similarly, substituting an equivalent amount of propionic anhydride for the acetic anhydride in the procedure of Example 16, 2 ^L -propionyl-erythromycin A-6,9-hemiketal-11,12-carbonate ester was prepared,

Example 17

S- ^ -acetyl -4 "-deoxy_-4" -oxo-erythromycin A-6,9-hemi-ketal-11,12-car bonate ester

To a suspension of 6.19 g of N-chlorosuccinimide in 150 ml of toluene and 50 ml of benzene, cooled to -5 C, 4.46 ml Diniethylsulfid added-

, 203 547

given. After stirring for 20 minutes, the resulting suspension is cooled to -25 ° C, and there are 12.4 g of 2 ^I -acetyl erythromycin A-6,9-hemiketal-11,12-carbonate ester, partially dissolved in 80 ml of toluene , added dropwise. The temperature, which is maintained between -19 C to -25 C during the addition, is kept at -25 C for 2 hours. At the end of this period, 6.79 ml of triethylamine are added all at once. The cooling bath is removed and the temperature is allowed to rise to -10 ° C. The reaction mixture is then poured into water and the aqueous phase is adjusted to 8.4 to 8.4. The organic layer is separated, dried over sodium sulfate and concentrated under vacuum to a white foam (14.0 g), admixing the residue with diethyl ether causes the foam to crystallize, filtering off and drying the product gives 11.3 g of crystalline material with F 212-213.5 ° C.

NMR (<r, CDCl ₃ ): 5.26 (iH) t, 3.36 (3H) s, 2.30 (6H) s, 2.13 (3H) s,

1.63 (3H) s and 1.50 (3H) s.

In the same way, 2 ^t -propionyl-4 "-deoxy-4" -oxo-erythromycin A-6,9-hemiketal-11,12-carbonate ester is prepared according to the procedure of Example 17 using the ^2f -acetyl ester is replaced by an equivalent amount of 2 ^l -propionyl-erythroraycin-A-6,9-hemiketal-11, i2-carbonate ester.

Example 18

4 "-deoxy_ ~ 4" -oxo-er-thromycin-A-6-hemiketal-H, 12-parbonate ester

42.9. g 2 ^t ~ -acetyl 4 ^M -deoxy-4 "-oxo-erythrotnycin A 6,9-hemiketal 11,12-carbonate ester are added to 800 ml of methanol, and the resulting solution is stirred at room temperature for 72 hours. When Removal of the solvent in vacuo leaves 4 g of the product as a white foam and the remaining material is dissolved in about 100 ml of acetone, followed by careful addition of water

- ³¹ . - 203 547

to the point of precipitation. The resulting crystalline solid is stirred for 40 minutes, then filtered and dried to yield 34.2 g of the desired product, mp 186.5-188 ° C.

NMR (cT, CDCl _3): 5.66 (IH) t, 3.35 (3H) s, 2.35 (6H) s, 1.65 (3H) s

and f, 5i (3H) s.

Similarly, the same product is obtained when an equivalent amount of 2-propionyl-4- ^ll- deoxy-4'- ^t- oxo-terythromycin A-6,9-hemiketal-11,12-carbonate ester is used in this procedure instead of the 2'-acetyl ester is used.

Example ^ 9 ^:

4 "-amino-4--Deoxy_ ^lt erythromv _i cin-A-6 _i-hemiketal 9-'l'l 'i2-carbonate ester

To 189 g of 4 "-deoxy-4 ^ll -oxo-erythromycin A 6,9-hemiketal 11, 12-carbonate esters are in 1200 ml of methanol at room temperature with stirring 193 g of ammonium acetate was added, ^ alas is 5 minutes, the resulting solution cooled to about -5 C, and then it is treated with 13.4 g of 85 / Sigera sodium cyanoborohydride in 200 ml of methanol for a 45-minute addition period. The cooling bath is removed and the reaction mixture is stirred at room temperature overnight. The reaction mixture is concentrated in vacuo to a volume of 8OO ml and added to a stirred mixture of 18QO ml of water and 9OO ml of chloroform. The pH is above. 6.2 to 4.3 with 6N hydrochloric acid, and the chloroform layer is separated. The chloroform is combined with 1 liter of water and the pH is adjusted to 9.5. The organic phase is separated, dried over atriumsulfat and concentrated under reduced pressure to give 174 g of a white foam, the remaining material is dissolved in a mixture of 1 1 V / acer and 5OO ml of ethyl acetate, and the pH-Uert is set to 5.5. The Äthylace

-32- 203 547

being extracted after each pH adjustment with 500 ml of fresh ethylacetate 5,7und then adjusted 9.5 - tatschicht is separated, and the aqueous layer is adjusted to pH. The ethyl acetate extract at pH = 9.5 is dried over sodium sulfate and concentrated to dryness in vacuo to give 130 g of product. 120 g of the remaining Schauemes are dissolved in a Geraisch of 1 1 of water and 1 1 of methylene chloride. The pH of the aqueous layer is adjusted to 4.4; 4.9 and then set 9> 4, which is extracted after each adjustment with 1 1 of fresh methylene chloride. The methylene chloride extract at pH = 9 »4 is dried over sodium sulfate and concentrated under reduced pressure to give 32 g of the product as a white foam. Crystallization from 250 ml of acetone-water (1: 1, v / v) gives 28.5 g of the crystalline spimers.

NMR 100 Mz ((T ₁ CDCl ₃ ): 5.20 (iH) m, 3.37 (i, 5H) s, 3.34 (i, 5H) s,

2.36 (6H) s, 1.66 (3H) s and 1.41 (3H) s.

Example 20

Separation of 3-dimers of the 4 "-deoxy_-4" -amino-erythrorriy_cin-A ~ 6,9-hemiketal 11,12-carbonate ester

A high pressure liquid chromatography column (1.27 χ 9 era) filled with Gf 254 silica gel impregnated with formamide and eluted with chloroform is charged with 200 mg of starting material. A pressure of 16.9 atm is applied at a rate of 4.76 ecm / min and a fraction size of 10 ml is chosen. Fractions 14 to 21 and 24 to 36 are collected.

Fractions 14 to 21 are combined and concentrated to about 50 ml. 50 ml of water are added and the pH is adjusted to 9.0. The Chloroforraschicht is separated, dried over sodium sulfate and concentrated to give 106 rag of a white Schauemes

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become. The Vereiben with diethyl ether causes the crystallization of the foam. After einstundigem stirring at room temperature, the crystalline product is filtered off and gertocknet, whereby 31.7 mg of F 194-196 ⁰ C are obtained.

NMR 100 Mz ( (Tt CDCX ₃ ): 5.2 '+ (iH) t, 5.00 (iH) t, 3,' K) (3H) s,

2.40 (6H) s, 1.66 (3H) s and 1.40 (3H) s.

Fractions 24 to 36 are combined and worked up as before to give 47.1 mg of product as a white foam which is identical to the product obtained in Example 25.

Beispieljii

To a suspension of 11.1 g of 2'-acetyl - ^ - oxo-erythromycin-Ao ^ - hemiketal-11, i2-carbonate ester in 300 ml of isopropanol at room temperature, 10.7 g of ammonium acetate are added with stirring. After 5 minutes will be 74? mg of sodium cyanoborohydride in 130 ml of isopropanol over a period ^of 30 minutes, and the resulting reaction mixture is stirred overnight at room temperature. The pale yellow solution is poured into 1100 ml of water, then 400 ml of diethyl ether are added, the pH is adjusted to 4.5, and the ether layer is separated. The aqueous layer is basified to pH = 9.5 and extracted with chloroform (2 × 500 ml). The chloroform extracts are combined together, dried over sodium sulfate and concentrated to give 7.5 g of a yellow foam. Recrystallization of the residual material from diethyl ether gives 1.69 g of product which is retained along with the mother liquors.

The mother liquor is treated with 75 ml of water and the pH is adjusted to 5.0. The ether layer is replaced by 75 μl of fresh ether and the pH is adjusted to 5.4. The ether is replaced by ethyl acetate and the pH is increased to 10. The

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Basified aqueous layer is extracted with ethyl acetate (2 × 75 ml) and the first ethyl acetate extract is dried over sodium sulfate and concentrated to dryness. The remaining 1 »96 S foam are added to a mixture of 75 ml of V / asser and 50 ml of diethyl ether, and the pH is adjusted to 5.05. The ether is separated off and the aqueous layer is successively adjusted to pH = 5.4-; 6.0; Adjusted to 7.05 and 8.0, wherein after each pH adjustment with-50 ml of fresh diethyl ether is extracted. The pH is finally adjusted to 9 | 7 and the aqueous layer is extracted with 50 ml of ethyl acetate. The ether extract, obtained at pH = 6.0, is combined with 75 ml of water and the pH is adjusted to 9 ». set. The ether layer is separated, dried and concentrated in vacuo to give 460 mg of a white foam *. ,

NMR 100 Mz ((T, CDCl-): 5.20 (1H) t, 3.43 (2H) s, 3,4'O (iH) s, 2.38

(6H) s, 2.16 (3H) s, i, 7O (3H) s and 1.54 (3H).

The NMR data show that the product is the epimer of 2'-acetyl-4 "-deoxy- ^ -amino-erythromycin A o -heraiketal H, 12-carbonate ester.

The aforementioned 1.69 g are dissolved in a mixture of 75 ml of water and 75 ml of diethyl ether, and the pH is adjusted to 4.7. The ether is separated off and the aqueous layer is further extracted with fresh ether (75 ml) at pH = .5.05 and 5.4, and with ethyl acetate (2 x 75 ml) at pH = 9.7. The combined ethyl acetate extracts are dried over sodium sulfate and concentrated under reduced pressure to yield 1.26 g of a white foam. The crystallization of this remaining material gives 411 rag product Rait F. 193-196 ⁰ C (dec.). The mother liquor is concentrated to dryness and the residue is dissolved in hot ethyl acetate. The solution is allowed to stand at room temperature for over eight hours. The crystalline solid which precipitates is filtered off and dried.

-35- 203 547

net, giving 182 mg of additional product with mp 198-202 ⁰ C (dec.).

HMR 100 Mh ( (f, CDClO: 5.10 (1H) t, 3.3 ^ (2H) s, 3.3 O (iH) s, 2.30

(6H) s, 2.08 (3H) s, 1.62 (3H) s and 1.48 (3H) s.

The NMR data show that the product is the epimer of 2'-acetyl-4 "-deoxy-4" -amino-erythromycin A ~ 11, i2-carbonate ester.

Similarly, when repeating the procedure of Example 21 using 2 ^l -propionyl-4 "-deoxy-4" ~ oxo-erythrortiycin A-6,9-heraiketal-11,12-carbonate ester as starting material of the 2nd ^l -PropionyJJ ^tl-4-deoxy-4 "-araino-erythroRiycin A 6,9-hemiketat-11,12-carbonate ester .and of 2 · ^f propionyl-4 ^ll -deoxy- f ^{z lt} -amino-erythroinycin --A-11, i2- <carbonate ester.

Beisgiel_22

Sine solution of 400 rag 2 ^l-Acetyl-4 "-deoxy-4 ^ll -amino-erythromycin A. _{6-t-9-hemiketal} 11,12-carbonate ester in 20 ml of methanol is stirred overnight at room temperature. The 'reaction solution is poured into 100 ml of water, followed by the addition of 50 ml of ethyl acetate, the pH is adjusted to 9.5 and the organic phase is separated off, and the extraction is repeated with 50 ml of fresh ethyl acetate Aqueous ethyl acetate extracts are dried over sodium sulfate and concentrated to give 392 mg of a white foam, which is then crystallized after leaching with diethyl ether and glass rod scraping, and after standing at room temperature for 30 minutes, the crystalline solids are filtered off and dried 123 mg of product are obtained, the mother liquor is retained.The product is identical to the material prepared in Example 24 on the basis of NMR values.

-36- 203 547

NMR 100 Mh (cT, CDCl _3): 3.26 (3H) s, 2.32 (6H) s, 1.61 (3H) s and

1, Vf (3H) s.

The NMR data show that the crystalline product is a single epimer of 4 "-deoxy-4" -araino-erythromycin A-11, i2 ~ carbonate ester.

The residual mother liquor is concentrated in vacuo to give 244 mg of a white foam.

This product is identical to the citerial from Example 19.

The NMR * V / he te show that this product is a Geraisch of the epimers of 4 ⁿ -deoxy-4 ⁿ -amino-erythromycin A 6,9-hemiketal 11, i2 carboantester and also identical with the product of Example 19 is

Beispiel_23

In a similar manner as in the procedure of Example 22, the methanolysis of 2 ^l -propionyl-4 "-deoxy-4 ' ^l -amino-erythromycin A-6,9-hemicotal-11,12-carbonate ester gave the 4" -deoxy- 4- ^lt -amino-erythro! Nyein-A-11, 12-carbonate ester and the 4 "-deoxy-4" -amino-erythromycin A 6,9-herniketal-11,12-carbonatsster.

Example 24

8 g of the epimer mixture of 4 "-deoxy-4 -araino ^{ll-erythromycin} A 11,12-carbonate ester from the micht crystalline product of Example 19 50 ml of diethyl ether were. The product was dissolved brought by scratching with a glass rod for crystallization. After stirring for 20 minutes, the crystalline product was filtered off and dried to give 1.91 k product of mp 198.5-20O ⁰ C.

NMR 100 Mz (cf, CDCl _3): 3.26 (3H) s, 2.30 (6H) s, 1.61 (3H) s and

1.45 (3H) s.

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The NMR data show that the crystalline product is a single epimer of 4 "-deoxy-4" -amino-erythromycin A11, i2-carbonate ester and that it is identical to the ketone product of Example 22,

Beisgiel 25

1 g of the epimer of Example 24 is dissolved in 20 ml of acetone and heated at steam bath temperature until the boiling point is reached. 25 ml of water are added and the resulting solution is stirred at room temperature, after stirring for one hour, the precipitate which forms is filtered off and dried to give 581 mg of product of mp 147-149 ° C.

NMR 100 Mz (□, CDCl-): 5.12 (iH) d, 3.30 (3H) s, 2.30 (6H) s, 1.62

(3H) s and 1.36 (3H) s.

The NMR data show that the product is a single epimer of the 4 "-deoxy-4" -amino-rerythroraycin-A-6,9-heniiketal-11, i2-carbonate ester and with the epireren in fractions 24-36 of the Example 20 is identical.

Example 26 4j! Deoxy-4 "-amino-erythro-nycin-A

20 g of 4 ^ll -deoxy-4 "-oxo-erythromycin A, 31.6 g of ammonium acetate and 10 g of 10% palladium on charcoal in 200 ml of methanol are dissolved at ambient temperatures in a hydrogen atmosphere at an initial pressure of

3.5 atm shaken overnight. The spent catalyst \ iir \ filtered off and the filtrate is · evaporated in vacuo to dryness. The residue is partitioned between Viasser chloroform at pH 5.5. The aqueous layer is separated, the pH is on

9.6, and it is separated, and chloroform is added. The organic layer is separated over sodium sulfate

203 547

dried and concentrated to dryness under reduced pressure. The remaining white foam (19 g) is triturated with 150 ml of diethyl ether at room temperature for 30 minutes. The resulting solids are filtered off and dried to yield 9-4x5 g of a single epimer which is indistinguishable from the product in Example 10. ·

The diethyl ether filtrate is concentrated to dryness to give 6.89 g of the product consisting of the other spiro plus some impurities,

Beispiel_27

4 ⁿ -deoxy_-4 "-amino-erythromycin-A

2 g of 4 ^N -deoxy-4- ^rt- OEO-erythrotnycin-A, 3 »1 B ammonium acetate and 2.0 g of Ranay nickel in 50 ml of methanol are shaken at room temperature in a hydrogen atmosphere at an initial pressure of 3 l-5 atm overnight , An additional amount of 3 g of ammonium acetate is added to 2.0 g of Eanay nickel and the hydrogenation is continued for a further 5 hours. The solids are filtered off and the filtrate is concentrated to dryness in vacuo. The residue is added with stirring to a mixture of water-chloroform and the pH is adjusted from 6.4 to 5 -5. The aqueous phase is separated, the pH-Vi ^r ert is adjusted to 9 »6, and there is added fresh chloroform. The chloroform extract is separated, dried over sodium sulfate and concentrated under reduced pressure to give 1.02 g of product as a yellow foam. The predominant isomer has the opposite configuration in the 4 "position compared to the compound of Example 10.

 - »- 203 547

Example_28 ·

2'-acetyl-4 "-deoxy_-4" -amino-erythromycin B

To a solution of 4.5 g of 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin ~ B (US Patent 3,884,937) in 45 ml of isopropanol under a nitrogen atmosphere with stirring 4.66 g of dry Ammonium acetate added. After 10 minutes, 376 mg of sodium cyanoborohydride are added to the reaction mixture with 10 ml of isopropanol, and the reaction mixture is stirred overnight at room temperature. The light yellow solution is poured into 400 ml of water and the pH is adjusted to 6.0. The aqueous phase is treated at pH = 6 ;, 7; 7.5; 8th; 9 and 10 extracted using 250 ml of diethyl ether for each extraction. The extracts at pH = 8, 9 and 10 are combined and washed with 250 ml of fresh water. The separated, aqueous layer is washed with ether (1 × 100 ml) at pH = 7, ethyl acetate (1 × 100 ml) at pH = 7, ether (1 × 100 ml) at pH = 7.5, ethyl acetate (100 ml ) at pH = 7.5 and ethyl acetate (1 x 100 ml) at pH = 8.9 and 10. Add the ethyl acetate extracts at pH = 9 and 10, wash with saturated brine and dry over sodium sulfate. Removal of the solvent in vacuo gives a spiked mixture of the desired product in the form of a cream colored foam.

In a similar manner, 4 "-deoxy-4" -oxo-erythromycin-B is used to prepare the compound 4 "-deoxy-4" -amino-erythrotnycin-B.

Beisgiel_29

4 "-deoxy-4" -amino-erythromycin B

A solution of 4.34 g of 2 ^l-Acetyl-4 "-deoxy-4 ^lt -amino-erythromycin-B in 100 ml of methanol at room temperature over" stirred eight. The solvent is removed under reduced pressure and the residual foam is washed with a mixture of 100 ml of chloroform and 100 ml

203 547

Water is treated, the pH of the aqueous layer is adjusted to 9.5, and the organic layer is separated. The chloroform layer is treated with fresh water and the pH is adjusted to 4-, O. The pH of the acidic aqueous layer containing the product is adjusted gradually to 5, 6, 7, 8 and 9 by the addition of base, extracting with fresh chloroform at each pH. The extracts at pH = 6 and 7 contain most of the product, and they are combined and treated with fresh water at pH = 4. The aqueous layer is readjusted to pH = 5.6 and 7, extracting with fresh chloroform at each pH. The chloroform extract at pH = 6 is dried over sodium sulfate and concentrated to give the product as a mixture of epimers. _:

Sample .30

V-Deoxy-4 "-amino-erythr omj / cin-A-G ^ -hemi-ketal -11 ^ 12 -carbonates terasgaraginat

To a solution of 1.0 g of 4 ^tl -deoxy-4 "-amino ^l _J .erythromycin-A-6,9-hemi

Ketal 11,12-carbonate ester in 6 ml of acetone heated to 40 C, 20 ml of V / asser and then 175 rag of L-aspartic acid zugesezt. The GE-

-1

The mixture is heated for 1.5 hours at reflux temperature, then it is filtered while hot. The filtrate is concentrated by removal of the acetone and lyophilized to give 1.1 g of the desired product as a white solid.

Beispiel_3i

4 "-De ox ^ -4" araino-er ^ thronjrcin-A-6 ^ -hemi ketal-11 _Α 12-Γ3Γ-carbonate terdihydrochloride

7 to _f 58 g of 4 "-deoxy-V-amino-erythromycin A 6,9-heraiketal-ii, 12-carbonate ester in 50 ml dry ethyl acetate, 20 ml of 1N ethyl acetate

203 547

solution of hydrogen chloride, and the resulting solution is concentrated to dryness under reduced pressure. The residual material is triturated with ether and filtered to give the desired salt.

Following a similar procedure, the amine compounds of the invention are converted to their diacid addition salts.

Beis £ iel_32_

4 "-Deoxy_-V-amino-erythrom VCin-A-6,9-hemiketal _11-A-i2 carbanatesterhydrochlorid

The procedure of Example 31 is repeated except that 10 ml of a 1N ethyl acetate solution of hydrogen chloride are added. The solution is concentrated to dryness in vacuo and the remaining monohydrochloride salt is triturated with ether and filtered.

Following a similar procedure, the amine compounds of the invention are converted to their monoacid addition salts.

(T)

oc ^

OCH

OCH N (CH) R ₁ . ' ^{3 2}

(U

HO

N (CH ^)

I ν <

(ml

N (CHJ.

R ₂ O R ₃ O

It

, f * ''''

203 547

A ₀ N (CH) ₂

OCH

3 ·

Ia

OCH.

ΊΓα

HO "Hf !!

_Er ythro rnycin_ _g_

A-QH

B - H

-Mtf-

^% OCH

N-

Il O CH ₃ CH ₂ C

Cm)

-43-6 3 ¹ 547

N (CH)

X ^'32

ςχ

^x 'Och.

1 o

CH.

  ο

CH ₃ CH ₂ C " O CH ₃ CH ₂ C-

O CH CH C-

CH ₃ C-

N (CH ₃ )

"" OCH ₃

OIi CH ₃ C- O Il C-O CH ₃ O Il C- O O if Il c- C - CH ₃ C- CH ₃ CH ₂ O Il CH ₃ CH ₂ ^CH 3 ? CH ₃ CH ₂

Sehern a

203 547

N (CHJ ₂

SchQma 2

HO

R ₂ OiHn ₁

JT '

(CH ₃ I ₂

U; L; -Kvi / i ο,., ...

20 $ 547

Scheme 3

4 »

N (CH ₃₎ (CH ₃ I ₂

N (CH ₃ I ₂

HO

JL

-S3-547

Scheme 4

(X

NH ₄ OCOCH ₃ Na CNBH ^

Il

(R ₂ + R ₃ --C-) _D N (CH? J,

¹ ο,. ^ Κ ο

OCH

Il

ii

Claims (5)

j -42- 203 547 claim of invention; A process for the preparation of 4 - "- deoxy-A ^u -amino-erythromycin A compounds of formula III and pharmaceutically acceptable acid addition salts thereof, wherein R and Rp are each hydrogen or alkanoyl of 2 to 3 carbon atoms; is a hydrogen atom and R _? and R_, when taken together, is the radical O. Il -C, R ^{1 is} a hydroxyl group and R is a bond with the Kohlenstoffumenene to which R ^{1 is} connected or R ¹ JSiologischerstoff and R is hydrogen, with the proviso that when Rp is hydrogen R is hydrogen, dadurc h in that they are prepared by reduction of compounds of the formula I ⁿ in which R, R ¹ , R, Rp and R are as defined above and Y is NH, N-OH or N-OCOCH, - Res te, with the proviso that that when Y is not an NOH or NOCOCII ₃ radical, R forms a bond with the carbon atom to which R ^{f is} attached, wherein A) the above-mentioned reduction is carried out as a catalytic reduction and b) when Y represents an NH group generated in situ from said corrosive ketones of formula I "by condensation of said ketones with the ammonium salt of an alkanecarboxylic acid, the aforesaid reduction is carried out with a hydride or as a catalytic reduction and if so desired c) if R. or R _? Hydrogen and / or if R or R _? Alkanoyl they are converted to alkanoyl or hydrogen and d) Formation of the pharmaceutically acceptable acid addition salts. 2, according to item 1, characterized in that the abovementioned reduction by hydrogen is carried out in the presence of Ranay nickel, palladium on carbon or platinum oxide. 203 547 3. ^ learn after point 1, characterized d a - d u r cn that Y is an NH group and that an excess of an ammonium salt of an alkanecarboxylic acid, preferably ammonium acetate, is used. 4. The method according to item 3, characterized in that the reducing agent is sodium cyanoborohydride. 5 * ^ experience according to item 1, characterized in that Y is an M group which is generated in situ from the corresponding ketone of the formula I by condensation with the ammonium salt of an alkanecarboxylic acid with hydrogen and a hydrogenation catalyst. For this ... / ^ .... pages formulas