SE445223B - SET TO PREPARE 4 "-AMINO-ERYTHOMYLIN-A-DERIVATE - Google Patents

Description

7800270-6 _ Erythromycin § A -OH B -H The structural formula shows that this antibiotic is composed of three main parts, namely: a sugar fragment known as cladinos, a second sugar component, which contains a basic amino substituent and which is known as desosamine and a 14-membered lactone ring, which is referred to as erythronolide-A or -B or, as in the present context, the macrolide ring. The macrolide ring is numbered with numbers without superscripts, as on the other hand the numbering for desosamine and cladinos bears prime and second characters.

A large number of erythromycin derivatives have been prepared in an attempt to modify its biological and pharmacodynamic properties.

U.S. Pat. No. 3,417,077 discloses the reaction product of erythromycin and ethyl carbonate as a highly active antibacterial agent. U.S. Patent No. 5,884,903 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 significant research (British Patent Specification 1,100,504, Tetrahedron Ietters, 1645 (1967) and Croatica Chemica Acta, 59, 275 (1967)) and a dispute over its structural identity ( Tetrahedron Ietters, 157 (1970) and British Patent Specification 1,541,022).

Sulfonamide derivatives of erythromycylamine are described in U.S. Pat. No. 3,985,105 as useful antibacterial agents. Other derivatives are also described (Ryden et al., J. Med. Chem., 16, 1059 (1975) and Massey et al., J. Med. Chem., 17, 105 (1974)) and are reported to have antibacterial activity in vitro and in vitro. live. 3 "Vaoozvo-6 Summary of the Invention.

It has now been found that certain new 4 "-deoxy-4" -amino-erythromycin-A derivatives are excellent as antibacterial agents. These compounds are represented by the formulas: III and a pharmaceutically acceptable acid addition salt thereof, in which formulas each of R 1 and R 4 represents hydrogen or alkanoyl having 2-3 carbon atoms; R 2 represents alkanoyl having 2-3 carbon atoms; R 3 represents hydrogen; R 2 and R 3 together may represent the group -.-; R 3 and R 4 together may represent the group eL-; A is methyl; and B is ethyl. A preferred group of compounds in this class of chemotherapeutic agents are those of formula III. Particularly preferred within this group are those compounds in which R 1 and H 5 together represent - -.

A second preferred group of compounds within this class of antibacterial agents are those of Formula IV. In particular, within this group, those compounds in which 24 represents hydrogen and also those in which R 5 and R 4 together represent 0 -ö- are preferred.

The process of the invention is characterized by reducing compounds of formula 78 wherein Y 'is NH, N-OH or N-OCOCH 2; and Y "is NH, and wherein (a) the reduction was carried out as catalytic hydrogenation, when Y" is NH and Y 'is NH, N-OH or N-OCOCH 3; (b) the reduction is effected by hydride reduction, when Y 'and Y "are NH prepared in situ from the corresponding ketones by condensation of said ketone with an ammonium salt of an alkanoic acid, and, if desired, (c) R 1 and R 4, when they represent hydrogen, are converted to alkanoyl and / or R 1 and R 4, when they represent alkanoyl, are converted to hydrogen, (d) pharmaceutically acceptable acid addition salts are prepared.

The stereochemical designation of the substituents on the sugar components and the macrolide ring is, according to the present invention, with the exception of the epimerization in the 4 "position, when indicated, the same as for naturally occurring erythromycin-A.

Detailed Description of the Invention In accordance with the method used for the synthesis of the 4 "-deoxy-4" -amino-erythromycin-A antibacterial agents of the present invention, the following scheme is applied, starting from a 2 ' alkanoyl-erythromycin-A or a derivative thereof: and the selective oxidation of I 'and II' to I resp. II (Y = O) is the first in the process and comprises reacting the compounds I 'and II' with trifluoroacetic anhydride and dimethyl sulfoxide followed by the addition of a tertiary amine, such as triethylamine.

In practice, trifluoroacetic anhydride and dimethyl sulfoxide are first combined in a reaction-inert solvent at about -65 ° C. After 10-15 minutes, the alcohols I 'and II' are added at such a rate that the temperature is maintained at about -65 ° C and does not rise above -30 ° C. At a temperature above -30 ° C, the trifluoroacetic anhydride-dimethyl sulfoxide complex is unstable. The reaction temperature is maintained between -30 and -65 ° C for about 15 minutes and then lowered to about -70 ° C. A tertiary amine is added at once and the temperature of the reaction mixture is allowed to rise for 10-15 minutes. The reaction mixture is then treated with water and worked up.

Regarding the amounts of the alcohol components used per mole of alcohol substrate, it can be mentioned that one mole each of the trifluoroacetic anhydride and the dimethyl sulfoxide is required. Experimentally, it is advantageous to use a 1- to 5-fold excess of the anhydride and dimethyl sulfoxide to accelerate the reaction. The tertiary amine should be used in an amount corresponding to the molar amount of trifluoroacetic anhydride used.

The reaction-inert solvent should be one which substantially solubilizes the reactants and does not react to any great extent with either the reactants or the products formed. Since this oxidation is carried out at a temperature between -30 and -65 ° C, it is preferred that the solvent, in addition to the above properties, has a freezing point below the reaction temperature. Solvents or mixtures thereof which meet these requirements are toluene, methylene chloride, ethyl acetate, chloroform and tetrahydrofuran. Solvents which meet the above requirements but which have a freezing point above the reaction temperature can be used in small amounts in combination with one of the preferred solvents. The especially preferred solvent for this process is methylene chloride.

Preferred compounds prepared by this process are 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-A, 11,2'-diacetyl-4 "-deoxy-4" -oxo-erythromycin-A-6.9 -hemiketal and 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-A-6,9-hemiketal-11,12-carbonic acid ester.

The reaction time is not critical and depends on the reaction temperature and the reactivity of the starting materials. At temperatures between about -30 and about -65 ° C, the reaction is complete in 15-30 minutes.

Regarding the order in which the reactants are added, it is preferred that the trifluoroacetic anhydride be combined with the dimethyl sulfoxide, whereupon the required alcohol substrate is added. It is further suggested, as pointed out above, that the reaction temperature be kept below -30 ° C. This is in line with Omura and co-workers, J. Org. Chem., 41, 957 (1976).

The second process used for the preparation of intermediates for further processing into the useful antibacterial agents is represented by the following scheme: 7.7800270-6 in 'II and II' The second process represents an oxidation reaction in which the 4 "hydroxy substituent in the compounds of formula I 'and II', in which each of Ac and R, represents alkanoyl having 2-5 carbon atoms; R; represents hydrogen; and R 2 and R 3 together may represent - -, oxidized to a 4 "- deoxy-4 "-oxo-erythromycin A compound. The process involves the use of N-chlorosuccinic acid amide and dimethyl sulfide as oxidizing agents. In practice, these two reactants are first combined in a reaction-inert solvent at about 0 ° C. 10-20 minutes, the temperature is lowered to a value between 0 and -25 ° C and the alcohol substrate I '15 or II' is added, while said temperature is maintained.After 2-4 hours of reaction, a tertiary amine is added, e.g. is triethylamine, and the reaction mixture is hydrolyzed and worked up. For each mole of alcohol substrate, one mole each of the N-chlorosuccinic acid amide and dimethyl sulfide was used. Experimentally, it is advantageous to use a 1-20-fold excess of the succinic acid amide and sulfide to accelerate the reaction. The tertiary amine should be used in an amount corresponding to the molar amount of succinic acid amide used.

The gantenot reaction inert solvent is one which markedly solubilizes the reactants but does not react to any appreciable degree with either the reactants or the products formed. Since the reaction was carried out at a temperature between about 0 and about -25 ° C, it is preferred that the solvent has a freezing point below the reaction temperature in addition to the above properties. Among the solvents or mixtures thereof which meet these requirements may be mentioned toluene, ethyl acetate, chloroform, methylene chloride and tetrahydrofuran. Solvents which meet the above requirements but which have a freezing point above the reaction temperature can also be used in small amounts in combination with one or more of the preferred solvents. The especially preferred solvent is toluene-benzene.

The preferred compounds prepared by this process are 11,2'-diacetyl-4 "-deoxy-4" -oxo-erythromycin-A-6,9-hemiketal, 2'-acetyl-4 "-deoxy -4 "-Oxy-erythromycin-A-6,9-hemiketal-11, 12-carbonic acid ester and 2'-acetyl-4" -deoxy-4 "-oxo-erythromycin-A.

The reaction time is not decisive and depends on the concentration, reaction temperature and the reactivity of the reactants. At a reaction temperature between 0 and -25 ° C, the reaction time is about 2-4 hours. As noted above, it is preferred that the alcohol substrate I 'or II' be added to the premix of the succinic acid derivative and the dimethyl sulfide. Both of the processes described here are peculiar due to the selectivity of the oxidation, which takes place exclusively at the 4 "hydroxy substituent and leaves the other secondary alcohols in the molecule unaffected.

The useful 4 "-deoxy-4" -oxo intermediates of the formula 78 wherein each of R 1 and R 2 represents alkanoyl having 2-3 carbon atoms and R 3 is hydrogen, are prepared by a compound of the formula wherein Y represents O and R 1 alkanoyl having 2-3 carbon atoms is treated with an alkanoic anhydride (R 2 O) and pyridine.

In practice, the ketone I is contacted with an excess of the anhydride in pyridine as solvent. It is preferred to use the anhydride in the reaction in such a large excess as a 4-fold excess. The reaction is conveniently carried out at ambient temperature.

At these temperatures, the reaction time is between about 12 and about 24 hours. The alkanoyl group in the 2 'position of the intermediate ketones I (Y = O) and II is prepared by solvolysis, the 2'-alkanoyl-4 "-deoxy-4" -oxo-erythromycin- A related compound is stirred with an excess of methanol overnight at room temperature. By removing the methanol and, if necessary, carrying out a subsequent purification of the remaining product, compounds of formulas I (Y = O) and II are obtained, in which R 1 represents hydrogen. As pointed out above, the ketones of formulas I (Y = O) and II useful intermediates for the preparation of the 4 "-deoxy-4" -amino-erythromycin A antibacterial agents of the present invention of formulas III and IV. Preferred intermediates in this group are 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-A-6,9-hemiketal-11,2-carbonic acid ester and 4 "-deoxy-4" -oxo-erythromycin -A-6,9-hemiketal-11,12-carbonic acid ester.

Several synthetic routes can be used to prepare the antibacterial agents of formulas III and IV from the required ketones I (Y = O) and II. The 4 "-deoxy-4" -amino-erythromycin A compounds of formula III are prepared by condensing the ketones II with the ammonium salt of a lower alkanoic acid and subsequently reducing the imine formed in situ. By "lower alkanoic acid" in the present context is meant an acid having 2-4 carbon atoms.

In practice, a solution of the ketone II in a lower alkanol, for example methanol or isopropanol, is treated with the ammonium salt of a lower alkanoic acid, for example acetic acid, and the cooled reaction mixture is treated with the reducing agent sodium cyanoborohydride.

The reaction is allowed to proceed at room temperature for several hours, before being subjected to hydrolysis and the product is isolated.

Although one mole of ammonium alkanoate is required per mole of ketone, it is preferred to use an excess as large as a 10-fold excess to ensure complete and rapid formation of the imine.

Such excess quantities have been shown to have little detrimental effect on the quality of the product. 7800270-6 - 11 It is preferred that the reducing agent be used in an amount of about two moles of sodium cyanoborohydride per mole of ketone.

The reaction time varies with the concentration, the reaction temperature and the reactivity of the reactants. At room temperature, which is the preferred reaction temperature, the reaction is substantially complete after 2-3 hours.

When the lower alkanol solvent is methanol, as pointed out above, substantial solvolysis of any alkanoyl group in the 2 'position occurs. To avoid the removal of such a group, it is preferred to use isopropanol as the reaction solvent.

As noted above, ammonium acetate is preferred as the ammonium alkanoate for this reaction.

In isolating the desired 4 "-deoxy-4" -amino-erythromycin A derivatives from any non-basic by-products or starting materials, the basic nature of the final product is used. Accordingly, an aqueous solution is extracted in a range of gradually rising pH, so that neutral or non-basic materials are extracted at a lower pH and the product at a pH above 5. The extraction solvents, either ethyl acetate or diethyl ether, are washed with brine. , dried over sodium sulphate and the product obtained is recovered by removing the solvent.

Further purification, if required, can be accomplished by column chromatography on silica gel according to known procedures.

As noted above, solvolysis of the 2'-alkanolyl group in the appropriate 2'-alkanolyl-4 "-deoxy-4" -amino-erythromycin A derivative can be accomplished by allowing a methanol solution of said compound to stand overnight at ambient temperature.

It is to be noted that amines related to both formula III and formula IV are formed during the reductive amination of ketones of formula II, in which R 2 and R 3 together represent the group -ë- and R 1 alkanoyl having 2-3 carbon atoms or hydrogen . This is illustrated in the following reaction scheme: 78ÛU27Û-6 n NH4ococH3 NacNBH3 o H = -C-) ON (cH3) 2 'å' z 0 CH3 3 I o CH3 _ l¶H¿ Écn III R / Ä 3, N (R3 + R4 = -C ") The amine products III and IV represented above are conveniently separated by selective crystallization in diethyl ether. Recrystallization of a mixture of the represented compounds III and IV in acetone-water induces the formation of half ketal in the amine IV, resulting in isolation of III as the only product.

The first direct synthetic route to the amine compounds IV is the same as discussed above and involves condensation of the ketone I with an ammonium alkanoate followed by reduction of the imine formed imine with sodium cyanoborohydride. Compounds of formula IV, in which R 1, R 3 and R 4 have the meanings given above, are also prepared by reduction of the above-mentioned imine using hydrogen and a suitable hydrogenation catalyst.

Experimentally, the appropriate ketone (I) in a lower alkanol, for example methanol or isopropanol, is treated with ammonium salt of a lower alkanoic acid, for example acetic acid, and the hydrogenation catalyst and mixture are shaken in a hydrogen atmosphere until the reaction is substantially decayed.

Although one mole of ammonium alkanoate is required per mole of ketone, it is preferred to use an excess up to a 10-fold excess to ensure complete and rapid imine formation. Such excess quantities have been shown to have little detrimental effect on the quality of the product.

The hydrogenation catalyst can be selected from a large number, but Éaney nickel and 5-10 ß palladium on charcoal are preferred. These catalysts can be used in varying amounts depending on how quickly the reaction is to be completed. Amounts between 10 and 200% by weight can be used effectively.

The hydrogen pressure in the hydrogenation vessel also affects the reaction rate. It is preferred for convenient reaction time that the initial pressure be 5 # 5 kPa. It is also preferred for convenience that the reduction be carried out at ambient temperature.

The reaction time depends on a number of factors, e.g. temperature, pressure, concentration and reactivity of the reactants. Under the above preferred conditions, the reaction is complete in 12-24 hours.

The product is isolated by filtering off the spent catalyst and removing the solvent in vacuo. The residue is then treated with water and the product is isolated from non-basic materials by extraction of the basic product with water of varying pH in the manner described above.

As pointed out above, when using methanol as the lower alkanol solvent, significant solvolysis of any alkanoyl group in the 2 'position occurs. To avoid the removal of such a group, it is preferred to use isopropanol as the reaction solvent.

The second synthetic route to 4 "-deoxy-4" -amino-erythromycin-A antibacterial agents of formula IV comprises a first conversion of the ketones of formula I (Y = O) to an oxime or an oxime derivative, i.e. Y = N-OH and N-O-SCH5 and subsequent reduction of the oxime or derivative thereof.

The oximes of the ketones I (Y = O) are prepared by reacting said ketones with hydroxylamine hydrochloride and barium carbonate in methanol or isopropanol at room temperature. In practice, it is preferred to use an excess of hydroxylamine and as much as a 5-fold excess gives the desired intermediate in good yields.

By carrying out the reaction at ambient temperature and selecting an excess of hydroxylamine, the desired oxime derivative of 1-5 hours can be prepared. The barium carbonate was used in molar amounts, 10 M which are twice as large as the amount of hydroxylamine hydrochloride used. The product is isolated by adding the reaction mixture to water and then making the whole basic to pH 9.5 and extracting with a water-immiscible solvent, for example ethyl acetate.

Alternatively, the reaction mixture can be filtered and the filtrate evaporated in vacuo to dryness. The evaporation residue is then partitioned between water of pH 9.0-9.5 and a water-immiscible solvent.

Preparation of the O-aetoethyloxime compounds of formula I (Y = N-O-8CH 5) is accomplished by acetylation of the corresponding oxime.

Experimentally, one mole of the oxime is reacted with one mole of acetic anhydride in the presence of one mole of pyridine or triethylamine. The reaction is facilitated to complete, if an excess of the anhydride and pyridine is used and an excess of 50-40% is preferred. The reaction was best carried out in an aprotic solvent, for example benzene or ethyl acetate, at room temperature all night. After the reaction is completed, water is added, the pH is adjusted to 9.0 and the product is separated in the solvent phase.

Preferred oximes and oxime derivatives which are useful intermediates for the preparation of the antibacterial agents derived from 4 "-deoxy-4" -amino-erythromcin-A are e.g. 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-A-oxime, 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-AO-acetyl-oxime, 4 "-deoxy-4 "-oxo-erythromycin-A-oxime and 4" -deoxy-4 "-oxo-erythromycin-A-O-acetyloxime.

The reduction of the ketone derivatives (Y = N-OH or NO-3CH5) is effected by catalytic hydrogenation, whereby a solution of the oxime or derivative thereof in a lower alkanol, for example isopropanol, and a Raney nickel catalyst are shaken in a hydrogen atmosphere at an initial pressure of 6 .9 MPa at room temperature all night. Thereafter, the catalyst is filtered off and the solvent is removed from the filtrate, whereby the desired 4 "-deoxy-4" -amino-antibacterial agent of formula IV is obtained. If methanol is used as the solvent in this reduction, it is likely that a 2'-alkanoyl group is solvolyzed. To avoid this side reaction, isopropanol was used.

Among these antibodies of formulas III and IV derived from 4 "-deoxy-4" -amino-erythromycin-A, both epimers of 4 "-deoxy-4" -amino-erythromycin-A-6,9 are preferred. hemiketal-11,12-carbonic acid ester both epimers of 4 "-deoxy-4" -amino-erythromycin-A and both epimers of 4 "-deoxy-4" -amino-erythromycin-A-11,12-carbonic acid ester.

In utilizing the chemotherapeutic activity of the salts of the formulas III and IV of the present invention which form salts, it is, of course, preferred to use pharmaceutically acceptable salts. Although water insolubility, high toxicity or lack of crystallinity may make any particular salts unsuitable or less desirable for use as such in a given pharmaceutical application, water insoluble or toxic salts may be converted to the corresponding pharmaceutically acceptable bases by decomposition of the salt in the manner indicated above or may they are converted to any desired pharmaceutically acceptable acid addition salt.

Examples of acids which give pharmaceutically acceptable anions are hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid and sulfuric acid, phosphoric acid, acetic acid, lactic acid, citric acid, tartaric acid, succinic acid, maleic acid, gluconic acid and aspartic acid.

As pointed out above, the stereochemistry of the starting materials leading to the antibacterial agents of the present invention is the same as that of the natural material. The oxidation of the 4 "hydroxyl group to a ketone and the subsequent conversion of said ketone to the 4" amines allow the stereochemistry of the 4 "substituent to be changed from that of the natural product. Accordingly, when the compounds I (Y = O) and II is converted to amines in any of the ways described above, it is possible to form two epimeric amines, it is experimental to note that both of the epimeric amines are present in the final product in varying proportions depending on the choice of synthesis method. predominantly consists of one of the epimers, this can be purified by repeated recrystallization in a suitable solvent to a constant melting point.The other epimer, namely that present in minor amounts in the originally isolated solid material, is the predominant pro » It can be recovered from it in a known manner, for example by evaporation of the mother liquor and repeated recrystallization of the residue to a a product with a constant melting point.

Although said mixture of epimers can be separated in known manner, for practical reasons it is advantageous to use said mixture as it is isolated from the reaction mixture. However, it is often advantageous to purify the epimer mixture by at least one recrystallization in a suitable solvent, subject it to column or high pressure liquid chromatography, solvent distribution or trituration in a suitable solvent. Although such purification does not necessarily involve separation of the epimers, it removes such foreign materials as starting materials and unwanted by-products.

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

The novel 4 "-deoxy-H" -amino-erythromycin-A derivatives described herein exhibit in vitro activity against a variety of gram-positive microorganisms, for example Staphylococcus aureus and Strepttococcus pyogenes, and against certain gram-negative microorganisms, for example those with spherical or ellipsoidal shape (cooks). Their activity is easily demonstrated by in vitro testing against various microorganisms in brain-heart infusion medium using the usual dual serial dilution technique. Their in vitro activity makes them useful for topical application in the form of ointments, creams and the like, for sterilization purposes, for example for hospital utensils, and as antimicrobial agents for industrial use, for example for water treatment, slime control and as preservatives in paints and for wood.

For in vitro use, for example for topical application, it is often convenient to mix the selected product with a pharmaceutically acceptable carrier, for example a vegetable oil or a mineral oil or an emollient cream. They can likewise be dissolved or dispersed in liquid carriers or solvents, for example water, alcohol, glycols or mixtures thereof, or other pharmaceutically acceptable inert media, i.e. media which do not have a detrimental effect on the active ingredient. For such purposes, it is generally accepted to use concentrations of the active ingredients between about 0.01 and about 10% by weight based on the total mixture.

In addition, many compounds of the present invention and their acid addition salts are active against gram-positive and certain gram-negative organisms, for example Pasteurella multocida and Neisseria sicca, in vivo upon oral and / or parenteral administration to humans and animals. Their in vivo activity is more limited with respect to susceptible organisms and is usually determined by infecting mice of substantially uniform body weight with the test organism and subsequent oral or subcutaneous treatment of the experimental animals with the test compound. In practice, for example, in 10 mice, intraperitoneally suitably diluted cultures containing approximately 1 (1) times ID100 (the lowest concentration of organisms required to inactivate the experimental animals) are inoculated.

Control tests are performed simultaneously, with lower dilutions being inoculated on mice as a test for possible variations in the virulence of the test organisms. The test compound is administered 0.5 hours after inoculation and the administration is repeated 4, 24 and 48 hours later. Surviving mice are kept for four days after the last treatment and the number of survivors is recorded.

When used in vivo, these new compounds can be administered orally or parenterally, for example by subcutaneous or intramuscular injection in doses from about 1 mg to about 200 mg per kg of body weight and day. Favorable doses are between about 5 and about 100 mg / kg body weight and day and preferred doses between about 5 and about 50 mg / kg body weight and day.

Vehicles suitable for parenteral injection can be either aqueous, such as water, isotonic saline, isotonic dextrose solution, Ringer's solution or non-aqueous, such as fatty oils of vegetable origin (cottonseed oil, peanut oil, corn oil, sesame oil) and dimethyl sulfide. Aqueous vehicles which do not interfere with the therapeutic efficacy of the preparation and which are non-toxic in the volume or proportion used (glyorol, propylene glycol, sorbitol). In addition, such mixtures are suitable, from which solutions can be prepared before administration. Such mixtures may contain liquid diluents, for example propylene glycol, diethylene carbonate, glycerol, sorbitol, etc .; buffer substances, hyaluronidase, local anesthetics and inorganic salts, which additives give the desired pharmacological properties. These compounds can also be combined with various pharmaceutically acceptable, inert carriers, e.g. solid diluents, aqueous vehicles, non-toxic organic solvents in the form of capsules, tablets, lozenges, throat lozenges, dry mixes, suspensions, solutions, elixirs and parenteral solutions or suspensions. In general, the compounds are used in various dosage forms at concentrations between about 0.5 and about 90% by weight of the total mixture.

The following examples illustrate the invention, but this is of course not limited to the special measures and details set forth in the examples, and many variations may be made without departing from the spirit of the invention or exceeding the scope of the invention.

Example 1: 2'-Acetyl-4 "-deoxy-4" -oxyn-erythromycin-A. To 3 ml of methylene chloride and 0.528 ml of dimethyl sulfoxide, which were cooled to about -65 ° C and kept under a nitrogen atmosphere, was added 0.652 ml of trifluoroacetic anhydride. After about one minute, a white slurry formed, indicating the presence of the trifluoroacetic anhydride-dimethyl sulfoxide complex. To the slurry thus obtained was added dropwise a solution in 7 ml of methylene chloride of 1.0 g of 2'-acetyl-erythromycin-A-ethyl acetate, which is obtained by recrystallization of 2'-acetyl-erythromycin-A in ethyl acetate, \ I1 10 §O 55 40 7800270-6- 18 the temperature was kept at about -65 ° C. The mixture thus obtained was stirred for 15 minutes at about -60 ° C and then cooled to -10 ° C.

Triethylamine (1.61 ml) was quickly added to the reaction mixture and the cooling bath was removed. After stirring for 15 minutes, the solution was added to 10 ml of water and the pH of the aqueous phase was adjusted to 10.

The organic phase was separated, washed first with water (3: 10 ml) and then with brine (1x10 ml) and dried over sodium sulphate. Removal of the solvent under reduced pressure gave 929 mg of the crude product. This was recrystallized from methylene chloride-hexane to give 320 mg of the purified product, m.p. 105-108 ° C.

NHR (δ, CDCl 3): 5.28 (§H) s, 2.21 (6H) s and 2.05 (5H) s.

In the same manner, 2'-propionyl-4 "-deoxy-4" -oxo-erythromycin-A was prepared using 2'-propionyl erythromycin A-ethyl acetate as starting material.

Example 2: 4 "-Deoxy-4" -oxo-erythromycin-A. A solution of 4.0 g of 2 "-acetyl-4" -deoxy-4 "-oxo-erythromycin-A in 75 ml of methanol was stirred at ambient temperature for 20 hours. The solvent was removed in vacuo and the remaining white foam was recrystallized from methylene chloride-hexane, §, 4 g, m.p. 170.5-1.5 ° C. man (6, cnc15) = 5.56 (5205 and 2.55 (eine.

A product identical to the above was isolated when 2'-propionyl-# "- deoxy-4" -oxo-erythromycin-A was treated with methanol at room temperature.

Example 5: 2'-Acetyl-4 "-deoxy-" "-oxo-erythromycin-A. To a stirred solution of 13.7 g of 4 "-deoxy-4" -oxo-erythromycin-A in 100 ml of ethyl acetate was added 2.5 ml of acetic anhydride, and the reaction mixture thus obtained was stirred at room temperature for 2 hours. The solution was added to 100 ml of water and the pH of the aqueous phase was raised to 9.5 by adding 6N = NaOH. The organic phase was separated, dried over sodium sulfate and evaporated to give 14.5 g of a white foam which, after recrystallization from methylene chloride-hexane, was found to be identical with the product of Example 1.

Example 4: 2'-Acetyl-4 "-deoxy-4" -oxo-erythromycin-A-oxime. To 500 ml of methanol was added 10.8 g of 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-A, 1.94 g of hydroxylamine hydrochloride and 11.0 g of barium carbonate, and the suspension thus obtained was stirred at room temperature for 5 hours. 5 hours. The mixture was filtered and the filtrate was evaporated under reduced pressure. The remaining foam was taken up in ethyl acetate, which was then washed with water at pH 9.5. The organic phase was separated, dried over sodium sulfate and evaporated in vacuo to give 10.6 g of the desired product.

NHR (δ, CDCl 3): 5.55 (5H) s, 2.50 (6H) s and 2.06 (5H) s.

Example 5: 2'-Acetyl-4 "-deoxy-4" -oxo-erythromycin-A-O-acetyloxime. To a solution of 550 mg of 2'-acetyl-4 "-deoxy-H" -oxyn-erythromycin-A-oxime in 50 ml of ethyl acetate was added with stirring 64.2 μl of acetic anhydride and the reaction mixture was stirred overnight at room temperature. An additional 15.8 μl of acetic anhydride and 25.4 μl of triethylamine were added and stirring was continued for 4 hours. The reaction mixture was added to water and the pH was adjusted to about 9.0. The ethyl acetate phase was separated, dried over sodium sulfate and evaporated in vacuo to give 500 mg of the desired product.

NMR (δ, CDCl 3): 5.58 (5H) s, 2.25 (6H) s, 2.20 (5H) s, 2.05 (5H) s and 1.56 (5H) s.

The procedure described above was repeated but 2'-propionyl-4 "-deoxy-4" -oxo-erythromycin-A-oxime and 4 "-deoxy-4" -oxyn-erythromycin-A-oxime were used instead of 2'-acetyl ~ 4 "-deoxy-4" -oxo-erythromycin-A-oxime and the respective O-acetyl derivatives were obtained.

Example 61 2'-Acetyl-4 "-deoxy-4" -amino-erythromycin-A. A mixture of 14.0 g of 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-A-O-acetyloxime and 10 g of isopropanol-washed Raney nickel in 400 ml of isopropanol was stirred in a hydrogen atmosphere at an initial pressure of 6 .9 MPa all night at room temperature, the catalyst was filtered off and the filtrate was evaporated to a white foam. This was dissolved in 400 ml of isopropanol and combined with 50 g of new isopropanol washed Raney nickel catalyst. The hydrogenation was continued all night at room temperature and an initial hydrogen pressure of 6.9 Ma. The catalyst was filtered off and the filtrate was evaporated to dryness in vacuo. Nan thus obtained 8.1% of the desired product. Example Gel 7: The procedure described in Example 6 was repeated using the appropriate O-acetyloxime to prepare the following 4 "- amino-erythromycin-A analogs: SO 55 40 7800270-6 2 ° Example" 8: 4 "-Deoxy-4" -amino-erythromycin-A. A solution of 2.17 g of 2'-acetyl-4 "-deoxy-4" -amino-erythromycin-A in 50 ml of methanol was stirred at room temperature overnight. The solvent was removed under reduced pressure and the remaining foam was treated with a mixture of 50 ml of chloroform and 50 ml of water. The pH of the aqueous phase was adjusted to 9.5 and the organic phase was separated. The chloroform phase was treated with fresh water and the pH was adjusted to 4.0. The pH of the acidic aqueous phase containing the product was successively adjusted to 5; 6; 7; 8 and 9 by adding base, extracting at each pH with new chloroform. The extracts at pH 6 and 7 contained the bulk of the product and these extracts were combined and treated with fresh water at pH 4. The aqueous phase was readjusted to pH 5; 6 and 7 and extracted at each pH with new chloroform. The chloroform extract at pH 6 was dried over sodium sulfate and concentrated to give 249 mg of the product as an epimer mixture. mm (δ, δ 5.50 (ounces, 5.26 (ams, 2.30 (sz-ns and 1, + + 6 (5H))).

Similarly, 4 "-deoxy-4" -amino-erythromycin-A was prepared by methanol solvolysis of 2'-propionyl-4 "-deoxy-4" -amino-erythromycin-A.

Example 9: 4 "-Deoxy-4" -amino-erythromycin-A. To a solution of 5.0 g of 4 "-deoxy-4" -oxo-erythromycin-A in 50 ml of methanol kept under a nitrogen atmosphere and stirring was added 5.16 g of dry ammonium acetate. After 5 minutes, 188 mg of sodium oyanoborohydride were rinsed into the reaction mixture with 5 ml of methanol and the reaction mixture was stirred at room temperature overnight. The light yellow solution was poured into 300 ml of water and the pH was adjusted to 6.0. The aqueous phase was extracted at pH 6; 7; 7.5; 8; 9 and 10, using 125 ml of diethyl ether for each extraction. The extracts at pH 8; 9 and 10 were combined and washed with 125 ml of fresh water. The separated aqueous phase was extracted with ether (1x10 ml) at pH 7, with ethyl acetate (1x10 ml) at pH 7, with ether (1x100 ml) at pH 7.5, with ethyl acetate (1x10 ml) at pH 7.5 and with ethyl acetate (1x100 ml) at pH 8; 9 and 10. The ethyl acetate extracts at pH 9 and 10 were combined, washed with a saturated brine and dried over sodium sulfate. the solvent was removed in vacuo to give 50 mg of an epimeric mixture of the desired product as an ivory foam.

Example 10: 4 "-Deoxy-4" -amino-erythromycin-A (a single epimer). A solution of 10.0 g of the epimer mixture of 2'-acetyl-4 "-deoxy-4" -amino-erythromycin-A in 150 ml of methanol was stirred at room temperature under nitrogen for 92 hours. . The solvent was removed in vacuo and the residue was dissolved in a stirred mixture of 150 ml of water and 200 ml of chloroform. The aqueous phase was discarded and 150 ml of fresh water was added. The pH of the aqueous phase was adjusted to 5 and the chloroform phase was separated. The pH of the aqueous phase was then adjusted to 5.5; 6; 7; 8 and 9 and extracted after each adjustment with 100 ml of new chloroform. Chloroform extracts obtained at pH 6; 7 and 8 were combined, washed first with water and then with a saturated brine solution and dried over sodium sulfate. The solvent was removed under reduced pressure to obtain 2.9 g of an epimer mixture of 4 "-deoxy-1" -amino-erythromyin. A. A 1.9 g sample of the mixture was triturated with diethyl ether, whereby a small portion of the undissolved foam crystallized. The solids were filtered off and dried, yielding 67 mg of a single epimer of 4 "-deoxy-4" -amino-erythromycin-A, m.p. 140 DEG-140 DEG.

Example 11: 11,2'-Diacetyl-4 "-deoxy-4'-oxo-erythromycin-A-6,9-hemiketal.

A solution of 10 g of 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-A in 250 ml of pyridine was treated with 40 ml of acetic anhydride and the reaction mixture thus obtained was allowed to stand at room temperature for 10 days. The bulk of the solvent was removed in vacuo and the remaining concentrate was added to a mixture of 150 ml of water and 100 ml of chloroform. The pH of the aqueous phase was raised to 9.0 and the chloroform was separated, dried over sodium sulfate and evaporated to dryness. NHR (δ, CDCl 5): 5.55 (5H) s, 2.26 (6H) s, 2.10 (5H) s, 2.05 (5H) s and 1.55 (5H) s.

Example Gel 12: The procedure described in Example 11 was repeated but starting from the appropriate 4 "-deoxy-4" -oxo-erythromycin-A and required alkanoic anhydride for synthesizing the following compounds: uccuaøz 5.1 .. .L 9 0 3 0 9 I CH 3 CH 2g- CH 3 CH 2 C-CH 2 O 3 H CH 3 CH 2 -CH 3 C- 10 15 55 7800270-6 22 Example 15f 11-Acetyl-4 "-deoxy-4" -oxo-erythromycin-A-6,9-hemiketal.

A solution of 5.0 μl of 1,2'-diacetyl-4 "-deoxy-4" -oxo-erythromycin-A-9,6-hemiketal in 50 ml of methanol was stirred under a nitrogen atmosphere overnight.

The solvent was removed in vacuo to give the desired product (5.0 g) as a yellow foam.

NHR (δ, CDCl 5): 5.55 (5H) s, 2.51 (6H) s, 2.15 (5H) and 1.55 (5H) s.

In the manner set forth in this Example 15, the compounds of Example 12 were converted to 11-acetyl-4 "-deoxy-4" -oxo-erythromycin-A-6,9-hemiketal and 11-propionyl-4 "-deoxy-4" - oxo-erythromycin-A-6,9-hemiketal.

Example 14: 11-Acetyl-4 "-deoxy-4" -amino-erythromycin-A-6,9-hemiketal.

To a stirred solution of 4.4 μl of acetyl-4 "-deoxy-4" -oxo-erythromycin-A-6,9-hemiketal and 4.58 g of ammonium acetate in 75 ml of methanol was added 505 mg of 85% sodium cyanoborohydride. . After stirring at room temperature overnight, the reaction mixture was poured into 500 ml of water and the whole was added with 250 ml of chloroform. The pH of the aqueous phase was adjusted to 9.8 and the chloroform phase was separated. The aqueous phase was again extracted with chloroform and the chloroform extracts were combined, dried over sodium sulphate and evaporated to a white foam. This was dissolved in a stirred mixture of 125 ml of water and 125 ml of new chloroform and the pH was adjusted to 4.9. The chloroform was separated and discarded and the aqueous phase was adjusted to pH 5; 6; 7 and 8 and extracted after each pH adjustment with new chloroform. The extracts obtained from the aqueous phase at pH 6 and 7 were combined, washed with saturated brine and dried over sodium sulfate. By removing the solvent, 1.72 g of the desired product was obtained in the form of a white foam. The product was dissolved in the least amount of diethyl ether and the solution was then treated with hexane to turbidity. The crystalline product formed was filtered off and dried, 155 g, m.p. 204.5-206.5 ° C.

NNE (6, CDCl 5): 5.51 (2H) s, 5.28 (1H) s, 2.51 (6H) s, 2.11 (5H) s and 1.5 (5H) s.

Example 15: The procedure described in Example 14 was repeated but with isopropanol instead of methanol as the reaction solvent and with the appropriate 4 "-deoxy-4" -oxo-erythromycin-A to give the following compounds: ". N (CH3) 2 3 CH3 H. N 2 1 C83 ä fi; 0 0 II YI ChsC- C313 C- O 0 "I! case-c3cu2c- 0 0 ll U C33 CHZC- C113 CHZC- 0 0 N II _ cxacazc-cassc- Example 16: 2'-Acetylerythromycin-A-6,9-hemiketal-11,2-carbonic acid ester.

To a solution of 15.2 g of erythromycin A-6,9-hemiketal-11,12-carbonic acid ester (U.S. Patent No. 5,417,077) in 150 ml of benzene was added 1.8 ml of acetic anhydride and the reaction mixture was stirred at room temperature. for 1.5 hours. The solution was poured into 200 ml of water and the aqueous phase was basified to pH 9.0. The benzene phase was separated, dried over sodium sulfate and evaporated in vacuo to 15.5 white foam. Upon trituration with 50 ml of diethyl ether, the foam crystallized. By filtering off and drying the product, 1.2.6 g of pure product with a melting point of 224.5-228.5 ° C were obtained.

NHR (δ, CDCl 5) = 3.56 (5H) s, 2.50 (6H) s, 2.06 (5H) s and 1.61 (5H) s. The procedure described in Example 16 was repeated but using of an equivalent amount of propionic anhydride instead of acetic anhydride to give 2'-propionylerythromycin-Af6,9-hemiketal-11,12-carbonic acid ester.

Example 17: 2'-Acetyl-4 "-deoxy-4" -oxyn-erythromycin-A-6,9-hemiketal-11,12-carbon gas ester. To a suspension cooled to -5 ° C of 6.19 g of N-chlorosuccinic acid amide in 150 ml of toluene and 50 ml of benzene was added 4.46 ml of dimethyl sulfide. After stirring for 20 minutes, the resulting suspension was cooled to -25 ° C and 12.4 g of 2'-acetylerythromycin-A-6,9-hemiketal-11,12-carbonic acid ester partially dissolved in 80 ml of toluene were added. The temperature, which was kept between -19 and -25 ° C during the addition, was kept at -25 ° C for two hours, at the end of this period 6.79 ml of triethylamine was added the whole amount of The cooling bath was removed and the temperature was allowed to rise to -10 ° C. The reaction mixture was then poured into water and the aqueous phase was adjusted from pH 8.4 to pH 9.0 The organic phase was separated, dried over sodium sulfate and evaporated in vacuo to a white foam (1.4.0 g).

By triturating the residue with diethyl ether, the foam crystallized. By filtering and drying the product, 11.5% of a crystalline material with a melting point of 212-215.5 ° 0 was obtained.

NHR (δ, CDCl 5): 5.26 (1H) t, 5.56 (5H) s, 2.30 (6H) s, 2.15 (3H) s, 1.65 (arne and 1.50 (ams) .

The procedure described in Example 17 was repeated but with an equivalent amount of 2'-propionyl-erythromycin-A-6,9-hemiketal-11,12-carbonic acid ester instead of the 2'-acetyl ester. Nan thus obtained 2'-propionyl-4 "-deoxy-4" -oxo-erythromycin-A-6,9-hemiketal-11,12-carbonic acid ester.

Example 18: 4 "-Deoxy-4" -oxo-erythromycin-A-6,9-hemiketal-11,12-carbon-acid ester. To 800 ml of methanol was added 42.9 g of 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin-AG, 9-hemiketal-11,12-carbonic acid ester, and the solution thus obtained was stirred at room temperature for 72 hours. . After the solvent was removed in vacuo, 41 g of the product remained in the form of a white foam. This was dissolved in about 100 ml of acetone, after which water was carefully added to the precipitation point. The crystalline solid thus obtained was stirred for 40 minutes and filtered off and dried, yielding 54.2 g of the desired product, m.p. 185.5-188 ° C.

NN (δ, CDCl 3) = 5.56 (1H), 5.55 (5H) S, 3.55 (6H) S, 1.65 (5H) S and 1.51 ((H) s.

In the same manner, the same product was prepared when an equivalent amount of 2'-propionyl-4 "-deoxy-4" -oxo-erythromycin-A-6,9-hemiketal-11,12-carbonic acid ester was used in the procedure described above instead of 2 '-acetyl ester.

Example 19: 4 "-Deoxy-4" -amino-erythromycin-A-6,9-hemiketalyl, 12-carbonic acid ester. To 189 g of 4 "-deoxy-4" -oxo-erythromycin-A-6,9-hemiketal-11,1? -Carboxylic acid ester in 1200 ml of methanol was added at room temperature and with stirring_195 g of ammonium acetate. After 5 minutes the solution thus obtained was cooled. to about -5 ° C and then treated with 15.4 g of 85 3 sodium cyanoborohydride in 200 ml of methanol for an additional period of 45 minutes, the cooling bath was removed and the reaction mixture was stirred at room temperature overnight.The volume of the reaction mixture was 7800270-6 was reduced to 800 ml in vacuo and the reaction mixture was added to a stirred mixture of 1800 ml of water and 900 ml of chloroform, the pH was adjusted from 6.2 to 4.3 with GN-H01 and the chloroform phase was separated, the chloroform was combined with 1 liter of water and the pH was adjusted to 9.5 The organic phase was separated, dried over sodium sulfate and evaporated under reduced pressure to give a white foam, which was dissolved in a mixture of 1 liter of water and 500 ml of ethyl acetate. ch pH was adjusted to 5.5. The ethyl acetate phase was separated and the aqueous phase was successively adjusted to pH 5. and 9.5 and was extracted after each pH adjustment with 500 ml of fresh ethyl acetate. The ethyl acetate extract obtained at pH 9.5 was dried over sodium sulfate and evaporated in vacuo to dryness (150 °). Of the evaporation residue obtained in the form of a foam, 120 was dissolved in a mixture of 1 liter of water and 1 liter of methylene chloride, the pH of the aqueous phase was successively adjusted to 4.4; 4.9 and 9.4 and was extracted after each pH adjustment with 1 liter of new methylene chloride. The methylene chloride extract obtained at pH 4.9 was dried over sodium sulfate and evaporated under reduced pressure to give 32 g of the product as a white foam. Crystallization in 250 ml of a mixture of acetone and water (1: 1 by volume) gave 28.5 g of the crystalline epimers.

NNE 100 Mz (5, CDCl 3): 5.20 (1H) m, 5.57 (1.5H) s, 3.54 (1.5H) s, 2.56 (6H) s, 1.66 (5H) ) s and 1.41 (5H) s.

Example 20: Separation of 4 "-deoxy-4" -amino-erytxcngcinrA-6, Qdraniketal 11,12- 'kogäggæsterns ggnmnæmu In högtrydærmätsh al oxnatumzdkuingskohmni (13x90 uno pæmadrm fi ldsaküox al kael KE 259 ümmegmuatrm fi al nmæmd O fl xehnnat down khnx fi Onn, häIkks 200 mg of an EP al mæi al breathing 4 "« & xndr4Wem fi noorythrycnicAf6,9-hemiketal-11f12 was cerated at a rate of 4.76 cubic centimeters per minute and a fraction size of 10 ng was used. Fractions 14-21 and 24-36 were upsanüades.

Fractions 14-21 were combined and evaporated to about 50 ml. Water (50 ml) was added and the pH was adjusted to 9.0. The chloroform phase was separated, dried over sodium sulfate and evaporated to give 106 mg of a white foam. Upon trituration with diethyl ether, the foam crystallized. After stirring at room temperature for 1 hour, the crystalline product was filtered off and dried to give 51 mg with a melting point of 194-196 ° C.

NNE 100 NZ (6, CDCl 3): 5.24 (1H) d, 5.00 (1H) t, 3.40 (3H) s, 2.40 (6H) s, 1.66 (5H) s and 1 , 40 (5H) s.

Fractions 24-56 were combined and worked up as indicated above to give 47.1 mg of a white product. Foam, which was found to be identical to the material of Example 25.

Example Gel 21: To a suspension of 11,1 g of 2'-acetyl-4 "-deoxy-4" -oxyn-erythromycin-A-6,9-hemiketal-11,13-carbonic acid ester in 500 ml of isopropanol was added with stirring at room temperature 10 , 7 g of ammonium acetate.

After 5 minutes, 747 mg of sodium cyanoborohydride in 150 nl of isopropanol were added over the course of 50 minutes, and the reaction mixture thus obtained was stirred at room temperature overnight. The pale yellow solution was poured into 1100 ml of water, which was then added with 400 ml of diethyl ether. The pH was adjusted to 4.5 and the ether phase was separated. The aqueous phase was basified to pH 9.5 and extracted with chloroform (2x500 ml). The chlorophore extracts were combined, dried over sodium sulfate and evaporated to give 7.5 S of a yellow foam.

By recrystallization of the evaporation residue into diethyl ether, 1.69

The nitrate was treated with 75 ml of water and the pH was adjusted to 5.0. Ether phase exchange sum 75 ml of new ether and the pH was adjusted to 5.4.

The ether-exchanged ethyl acetate and the pH were raised to 10. The aqueous phase, which was basified, was extracted with ethyl acetate (2x75 ml) and the first ethyl acetate extract was dried over sodium sulphate and evaporated to dryness. The remaining foam (1.96 g) was added to a mixture of 75 ml of water and 50 ml of diethyl ether and the pH was adjusted to 5.05. The ether was separated and the aqueous phase was successively adjusted to pH 5.4; 6.0; 7.05 and 8.0 and extracted after each pH adjustment with 50 ml of new diethyl ether. The pH was finally adjusted to 9.7 and the aqueous phase was extracted with 50 ml of ethyl acetate. The ether extract, obtained at pH 6.0, was combined with 75 ml of water and the pH was adjusted to 9.7. The ether phase was separated, dried and evaporated in vacuo to give 460 mg of a white foam. mm 100 Hz (a, cnc15) = 5.20 (une, 3.45 (2105, 5.40 (nns, 2.58 (6H) s, 2.16)), fans, 1.70 (fans and 1.54 (5H).

NHR data show that the product consisted of the epimers of_gL; acetyl-4 "-deoxy-4" -amino-erythromycin-A-6,9-hemiketal-11,2-carbonic acid ester.

The above recrystallized residue (1.69 g) was dissolved in a mixture of 75 ml of diethyl ether and the pH was adjusted to 4.7. The ether was separated and the aqueous phase was further extracted with fresh ether (75 ml) at pH 5.05 and 5.4 and with ethyl acetate (2x75 ml) at pH 9.7. The combined ethyl acetate extracts were dried over sodium sulfate and evaporated under reduced pressure to give 1.26 g of a white foam which, by recrystallization, gave 411 mg of product, m.p. 193 DEG-196 DEG C. (decomposition). The mother liquor was evaporated to dryness and the residue was dissolved in hot ethyl acetate.

The solution was allowed to stand overnight at room temperature. The crystalline precipitate was filtered off and dried, yielding an additional 182 mg of product, m.p. 198-202 ° C (dec.).

NHR 100 M 2 (h, CDCl 5): 5.1C (1H) t, 5.54 (2H) s, 5.50 (1H) s, 2.50 (sms, 2.02, (ans, 1.62) sms and 1.48 (5H) s.

NHR data show that the product was the epimers of 2'-acetyl-4 "-deoxy-d" -amino-erythromycin-A-11,12-carbonic acid ester.

The procedure described in Example E1 was repeated but starting from 2'-propionyl-4 "-doxy-4" -oxo-erythromycin-A-6,9-hmiketal-11,13-carbonic acid ester and 2'-propionyl-H " -deoxy-4 "-amino-erythro-nino-A-6,9-hemiketal-11,1-carbonic acid ester and 2'-propionyl-4" -deoxy-1 "-amino-erythromycin-A-11,12- carbonic acid ester.

Example 23: A solution of 400 mg of β-acetyl-β-deoxy-α-amino-erythromycin-A-6,9-hemiketal-11,2-carbonic acid ester in 20 ml of methanol was stirred overnight at room temperature. The reaction solution was poured into 100 ml of water, then 50 ml of ethyl acetate were added. The pH was adjusted to 9.5 and the organic phase was separated. The extraction was repeated with 50 ml of fresh ethyl acetate. The combined ethyl acetate extracts were dried over sodium sulfate and evaporated to give 592 mg of a white foam which crystallized on trituration with diethyl ether and scraping with a glass rod. After standing at room temperature for 50 minutes, the crystalline solid was filtered off and dried to give 125 mg. Mother liquor was maintained. The product was found by HM to be identical to the material prepared according to Example 24.

NHR 100 MZ (e, cnc15) = 5.26 (5H) s, 2.32 (6H) s, 1.61 (5H) s and 1.44 (5H) s.

NNE data show that the crystalline product is a single epimer of 4 "-deoxy-4" -amino-erythromycin-A-11,12-carbonic acid ester.

The mixture was evaporated in vacuo to give 244 mg of a white foam. The product was found to be identical to the material of Example 19.

NHR data show that this product is a mixture of the epimers of 4 "-deoxy-4" -amino-erythromycin-A-6,9-hemiketal-11, 12-carbonic acid ester and identical to the product of Example 19.

Example Gel 25: In a manner similar to that described in Example 22, methanolysis of 2'-propionyl-4 "-deoxy-4" -amino-erythromycin-A-6,9-hemiketal-11, 12-carbonic acid ester and 4 "-deoxy-4" -amino-erythroglycine-A-11,12-carbon gas ester and 4 "-deoxy-4" -amino-erztroglcin-A-6,2-hemiketal-11,12-carbonic acid ester were obtained. Example ZAÄ Eight grams of the epimer mixture of 4 "-deoxy-4" -amino-erythromycin-A-11,12-carbonic acid ester from the non-crystalline product of Example 19 was dissolved in 50 ml of diethyl ether. Crystallization of the product was induced by scraping with a glass rod. After stirring for 20 minutes, the crystalline product was filtered off and dried, yielding 1.91 g, m.p. 198.5-200 ° C. nns 100 Hz (s, cno15) = 5.26 (5H) s, 2.50 (6H) s, 1.61 (3H) s and 1.45 (5H) e.

NHR data show that the crystalline product is a single epimer of 4 "-deoxy-4" -amino-erythromycin-A-11,12-carbonic acid ester and identical to the ketone product of Example 22.

Example 25: One gram of the epimer of Example 24 was dissolved in 20 ml of acetone and the solution was heated at a water vapor bath temperature until the boiling point was reached. Water (25 ml) was added and the solution thus obtained was stirred at room temperature. After stirring for 4 hours, the precipitate was filtered off and dried, yielding 581 mg, m.p. 47-149 ° C.

NER 100 Mz (5, CD015): 5.12 (1H) d, 5.50 (5H) s, 2.50 (6H) s, 1.62 (5H) s and 1.56 (5H) s.

NHR data show that the product is a single epimer of 4 "-deoxy-4" -amino-erythromycin-A-6,9-hemiketal-11,12-carbonic acid ester and identical to the epimer in fractions 24-36 in Example 20.

Example 26: 4 "-Deoxy-4" -amino-erythromycin-A. Twenty grams of 4 "-deoxy-#" -oxo-erythromycin-A, 51.6 ammonium acetate and 10 g of 10% palladium on charcoal in 200 ml of methanol were shaken at ambient temperature overnight in a hydrogen atmosphere with an initial pressure of 545 kPa. The spent catalyst was filtered off and the filtrate was evaporated to dryness in vacuo. The evaporation residue was partitioned between water and chloroform at pH 5.5. The aqueous phase was separated, the pH was adjusted to 9.6 and chloroform was added. The organic phase was separated, dried over sodium sulfate and evaporated to dryness under reduced pressure. The remaining white foam (19 g) was triturated with 150 ml of diethyl ether at room temperature for 50 minutes. The solid thus obtained was filtered off and dried, yielding 9.45 g of a single epimer which could not be separated from that of Example 10.

The diethyl ether filtrate was evaporated to dryness to give 6.89 g of a product consisting of the second epimer plus some impurities.

Example 27: 4 "-Deoxy-4" -amino-erythromycin-A. Two grams of 4 "-deoxy-4" -oxo-erythromycin-A, 5.1 g of ammonium acetate and 2.0 g of Raney nickel in 50 ml of methanol were shaken at room temperature in a hydrogen atmosphere overnight at a initial pressure of 545 kPa. An additional 5.16 g of ammonium acetate and 2.0 g of Raney nickel were added and hydrogenation was continued for a further 5 hours. The solid was filtered off and the filtrate was evaporated to dryness in vacuo. The evaporation residue was added with stirring to a mixture of water-chloroform and the pH was adjusted from 6.1 to 5.5. The aqueous phase was separated, the pH was adjusted to 9.6 and new chlorophore was added. The chloroform extract was separated, dried over sodium sulfate and evaporated under reduced pressure to give 1.02 g of the product as a yellow foam.

The predominant isomer had the opposite configuration at 4 "compared to the compound of Example 10.

Example 28: 2'-Acetyl-4 "-deoxy-4" -amino-erythromycin-B. To a solution of 4.5 2'-acetyl-4 "-deoxy-#" -oxo-erythromycin-B (U.S. Pat. No. 3,6S,905) in 45 ml of isopropanol under a nitrogen atmosphere was added with stirring 4.66 5 dry ammonium acetate. After 10 minutes, 576 mg of sodium oyanoborohydride were purged into the reaction mixture with 10 ml of isopropanol and the reaction mixture was stirred at room temperature overnight. The light yellow solution was poured into 400 ml of water and the pH was adjusted to 6.0. The aqueous phase was extracted at pH 6; 7; 7.5; 8; 9 and 10, using 250 ml of diethyl ether for each extraction. The extracts at pH 8; 9 and 10 were combined and washed with 250 ml of fresh water. The separated aqueous phase was extracted with ether (1 x 100 ml) at pH 7, with ethyl acetate (1 x 100 ml) at pH 7, with ether (1 x 100 ml) at pH 7.5, with ethyl acetate (1 x 100 ml) at pH 7. And with ethyl acetate (1x10 ml) at pH 8; 9 and 10. The ethyl acetate extracts prepared at pH 9 and 10 were combined, washed with a saturated saline solution and dried over sodium sulfate. The solvent was removed in vacuo to give an epimer mixture of the desired product in the form of a creamed foam.

Similarly, 4 "-deoxy-4" -amino-erythromycin-B was prepared from 4 "-deoxy-4" -oxo-erythromycin-B.

Example 29: 4 "-Deoxy-4" -amino-erythromycin-B. A solution of 4,54 2'-acetyl-4 "-deoxy-4" -amino-erythromycin-B in 100 ml of methanol was stirred at room temperature overnight. The solvent was removed under reduced pressure and the remaining foam was treated with a mixture of 100 ml of chloroform and 100 ml of water. The pH of the aqueous phase was adjusted to 9.5 and the organic phase was separated.

The chloroform phase was treated with fresh water and the pH was adjusted to 4.0.

The acidic aqueous phase containing the product was successively adjusted to pH 5; 6; 7; 8 and 9 by addition of base and extracted at each temperature with new Éloroform. The extracts at pH 6 and 7 contained the bulk of the product and these extracts were combined and treated with fresh water at pH 4. The aqueous phase was readjusted again to pH 5; 6 and 7 and extracted at each pH with new chloroform. The chloroform extract at pH 6 was dried over sodium sulfate and evaporated to give the product as an epimer mixture.

Example 50: 4 "-Deoxy-4" -amino-erythromycin-A-6,9-hemiketal-11,12-carbonic acid ester aspartate. To a solution of 1.0 g of 4 "-deoxy-4" -amino-erythromycin-A-6,9-hemiketal-11,12-carbonic acid ester in 6 ml of acetone heated to 40 ° C was added 20 ml of water and then l? 5 ml Ifasparagin ~ acid. The mixture was heated to reflux for 1.5 hours and then filtered hot. The filtrate was concentrated by removing the acetone and then lyophilized to give 1.1 g of the desired product as a white solid.

Example 31: 4 "-Deoxy-4" -amino-erythromycin-A-6,9-hemiketal-11,12-carbonic acid ester dihydrochloride. To 7.58 5 4-deoxy-H "-amino-erythromycin-A-6,9-hemiketal-11,12-carbonic acid ester in 50 ml of dry ethyl acetate was added 20 ml of a 1-normal ethyl acetate solution of H01 and the resulting the solution was evaporated to dryness under reduced pressure. The evaporation residue was triturated with ether and filtered off to give the desired salt.

Similarly, the amine compounds of the present invention were converted to their diacid addition salts.

Example 52: 4 "-Deoxy-4" -amino-erythromycin-β6,9-hemiketal-11,12-carbonic acid ester hydrochloride. The procedure described in Example 31 was repeated with the change that 10 ml of a 1-normal ethyl acetate solution of H01 was added. the solution was evaporated to dryness in vacuo and the remaining monohydrochloride salt was triturated with ether and filtered off.

Similarly, the amine compounds of the present invention were converted to their monoacid addition salts.

Claims (5)

A method of preparing 4 "-amino-epimeric compounds of the formula N (CH 3) 2 O CH 3 and O C83 = - myN fl 2 OCBB and pharmaceutically acceptable acid addition salts thereof, in which formulas were and one of R 1 and R 4 represents hydrogen or alkanoyl having 2-3 carbon atoms; R 2 alkanoyl having 2 to 3 carbon atoms; R 3 hydrogen; R 2 and R 3 together may form -C-; O R 3 and R 4 together may form -C- ; A is methyl; and B is ethyl, characterized in that compounds of the formula N (CH3) CH3 CH3 * \ || \ Y CH3 wherein ¥ 'is NH, N-OH or N-OCOCH3 And Y "is NH, and wherein (a) the reduction was carried out as catalytic hydrogenation, when Y" is NH and Y 'is NH, N-OH or N-OCOCH 3; (b) the reduction is effected by hydride reduction, when Y 'and Y "are NH prepared in situ from the corresponding ketones by condensing said ketone with an ammonium salt of an alkanoic acid, and, if desired, (c) R 1 and R 4, when representing hydrogen, converted to a1k an ° Y1 °° h / or R1 °° h R4, when they represent alkanoyl, are converted to hydrogen; (d) pharmaceutically acceptable acid addition salts are prepared. 2. A process according to claim 1, characterized in that Y "is NH and Y 'is NH, N-OH or N-OCOCH3 and that the reduction is catalytic reduction. 3. A method according to claim 2, characterized in that the reduction is effected with hydrogen in the presence of Raney nickel, palladium on charcoal or platinum oxide. 4. A method according to claim 1, characterized in that each of Y 'and Y "is NH and that an excess of an ammonium salt of an alkanoic acid, preferably ammonium acetate, is used. 5. A method according to claim 4, characterized in that the reducing agent is sodium cyanoborohydride.