KR820001217B1 - Process for preparing semi-synthetic 4-erythromycin a derivative - Google Patents

Abstract

Title compds. [III(or IV); R1 = R4 = H, C2-3 alkanoyl; R2 = C2-3 alkanoyl; R3 = H , useful as fungicides, were prepd. by the reduction of I(or II; Y1 = NH, N-OH, N-OCOCH3; Y" = NH). Thus, 14.0 g 2'-acetyl-4"-deoxy-4"-oxoerythromycin A O-acetyl oxime was reduced with 60 g Ra-Ni catalyst at room temp, 1000 psi in 400 ml iso-propanol to give 8.1 g 2'-acetyl-4"-deoxy-4"-amino-erythromycin A.

Description

Method for preparing a semisynthetic 4 "-erythromycin A-derivative

The present invention relates to a 4 "-deoxy-4" -amino-erythromycin A derivative of the following structural formulas (III) and (IV) useful as an antibacterial agent and a method for preparing a pharmaceutically toxic acid addition salt thereof.

이고, R₃ 및 R₄는 함께일 경우

이다.R 'and R' are each hydrogen or alkanoyl having 2 to 3 carbon atoms, R2 is alkanoyl having 2 to 3 carbon atoms, R₃ is hydrogen, or R₂ and R₃ together

When R₃ and R₄ are together

to be.

인 화합물이다.Among the compounds, preferred compounds as chemotherapeutic agents are compounds of formula (III). Particularly preferred among these compounds is that R2 and R₃ together

Phosphorus compound.

A second preferred compound of the above compounds which is useful as an antimicrobial agent is the compound of formula IV.

인 화합물이다.Particularly preferred among these compounds is a compound wherein R₄ is hydrogen, and R₃ and R₄ together

Phosphorus compound.

Erythromycin is an antibiotic that is formed during incubation of a strain of Streptoices erythrois in a suitable medium, as described in US Pat. No. 2,653,899. Erythromycin is produced in two forms, A and B, and is represented by the following structural formula.

From the above structural formula, it can be seen that the antibiotic consists of three main parts. Sucrose fraction known as cladinose, a second sucrose site containing a basic amino substituent, known as desosamine, and a 14-membered lactone or microlide ring known as erythrolide A or B. The number of the microride ring is just a number, the desosamine is represented by prime (') and the cladinose is represented by double prime (").

Many derivatives of erythromycin have been prepared for the purpose of improving biological and pharmaceutical properties.

US Pat. No. 3,417,077 describes reaction products of erythromycin and ethylene carbonate as excellent antibacterial agents, and US Pat. No. 3,884,903 describes 4 "-deoxy-4" -oxo-erythromycin A and B derivatives useful as antibiotics. It is.

, 273(1967)〕, 그리고 그의 구조에 관해서 논쟁되어 왔다〔참조 : Tetrahedron Letters, 157(1970) 및 영국특허 제1,341,022호〕. 에리트로마이실아민의 설폰아미드 유도체는 항균제로서 유용하며 미합중국 제3,983,103호에 기술되어 있다.Erythromylamine, a 9-amino derivative of erythromycin A, has been steadily studied [see, eg, British Patent No. 1,100,504, Tetrahedron Letters 1645 (1967) and Croatica Chemica Acta,

, 273 (1967), and their structure (Tetrahedron Letters, 157 (1970) and British Patent No. 1,341,022). Sulfonamide derivatives of erythromyylamine are useful as antibacterial agents and are described in US Pat. No. 3,983,103.

Other derivatives have also been reported to have antimicrobial activity in vivo and ex vivo [see Ryden et al. J. Med. Chem. 16, 1059 (1973) and Massey et al. J. Med. Chem., 17, 105 (1974)].

Compounds used as intermediates to prepare the structural formula (III) or (IV) antimicrobial agents are represented by the following structural formula (I) and (II).

-CH₃이고, R₃는 수소이고,In the structural formula R 'is hydrogen or alkanoyl having 2 to 3 carbon atoms, R2 is alkanoyl having 2 to 3 carbon atoms, Y is N-OH or NO-

-CH₃, R₃ is hydrogen,

이다.When R₂ and R₃ are together

to be.

Preferred intermediates are compounds of formula (I), particularly when R 'is hydrogen or acetyl.

The second preferred compound is of the formula (II) and is particularly preferred when R ₁ is hydrogen or R ₁ is acetyl.

Compounds of the following formula (I) or (II) to be used as starting materials in the present invention are dimethyl at -30 to -65 ° C in the presence of a solvent inert to the reaction of the following formula (I ') or (II') Prepared by reacting with each of 1 mole of sulfoxide and trifluoroacetic anhydride followed by contact with at least one mole of triethylamine.

In the structural formula, Ac and R2 are each alkanoyl having 2 to 3 carbon atoms, R₃ is hydrogen,

이다.R₂ and R₃ together

to be.

Preference is given to oxidizing the compounds of formulas (I ') and (II') in an inert reaction solvent such as methylene chloride.

Another method for preparing a compound of formula (I) or (II) used as a starting material in the present invention is to prepare a compound of formula (I ') or (II') in the presence of a solvent inert to the reaction. After reacting with 1 mole each of N-chlorosuccinimide and dimethyl sulfide at 25 ° C., it is reacted with one or more moles of triethylamid.

In the above structural formula

Ac and R2 are each alkanoyl of 2 to 3 carbon atoms,

R₃ is hydrogen,

이다.R₂ and R₃ together

to be.

Preferred among the reactions is the use of benzene and toluene as inert-reaction solvents.

Throughout the present invention, the chemical meaning of the sucrose and macrolide cyclic substituents is that of naturally occurring erythromycin (except epimerization at the 4 "-position).

Also included in the present invention are derivatives of erythromycin B corresponding to the derivatives of structural formulas (I) and (II).

Erythromycin B compounds are also useful as intermediates and are prepared by the same synthetic procedures as erythromycin A compounds. Erythromycin B intermediates are also converted to erythromycin B amines corresponding to the compounds of formulas (III) and (IV) of the present invention according to the methods described above. Erythromycin B amines are also useful as antibacterial agents.

The starting material used to prepare 4 "-deoxy-4" -amino-erythromycin A of the present invention is 2'-alkanoyl-erythromycin A, or a derivative thereof using the starting material in the following scheme To produce accordingly.

Selective oxidation of the compounds of the formulas (I ') and (II') into the compounds of the formulas (I) and (II) (Y = 0) respectively is the first step of the present invention. II ') is reacted with trifluoroacetic anhydride and dimethylsulfoxide followed by addition of a tertiary amine such as triethylamine.

In practice, trifluoroacetic anhydride and dimethyl sulfoxide are first mixed at about −65 ° C. in the presence of a reaction-inert solvent and after 10-15 minutes the alcohol (I ′) or (II ′) is kept at -65 ° C. Do not add more than -30 ° C. At temperatures above -30 ° C, the trifluoroacetic anhydride-dimethylsulfoxide complex is not stable. The reaction temperature is maintained at -30 to -65 ° C for 15 minutes and then lowered to -70 ° C. Tertiary amine is added at once and the reaction is warmed for 10 to 15 minutes, then the reaction mixture is treated with water and finished.

In terms of the amount of reactants, one mole each of trifluoroacetic anhydride and dimethylsulfoxide per mole of alcohol used is required. Experimentally, it is preferable to use an anhydride and dimethyl sulfoxide in 1 to 5 times excess to complete the reaction quickly. The tertiary amine used should correspond to the molar amount of trifluoroacetic anhydride used.

The reaction-inert solvent used in this reaction must be capable of dissolving the reactant and not reacting with the reactant or the formed product. This oxidation reaction is carried out at -30 to -65 ° C. In addition to the above characteristics it is preferable to use a solvent having a freezing point below the reaction temperature.

Such solvents or mixtures thereof include such as toluene, methylene chloride, ethyl acetate, chloroform or tetrahydrofuran.

Solvents that meet the above requirements or have a freezing point or higher reaction temperature can be used in small amounts by mixing with one of the preferred solvents. Particularly preferred solvents for this reaction are methylene chloride.

Preferred compounds prepared by this reaction are 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin A, 11,2'-diacetyl-4 "-deoxy-4" -oxo-erythromycin A 6,9-hemictal and 2'-acetyl-4 "-deoxy-4" -oxo-erythymycin A 6,9-hemictal 11, 12-carbonate ester.

The reaction time is not limited and depends on the reaction temperature and the intrinsic reactivity of the starting reactants.

The reaction at a temperature of -30 to -65 ° C. is completed within 15 to 30 minutes.

In the addition of the reactants, it is preferred to add the necessary alcohols after mixing trifluoroacetic anhydride with dimethylsulfoxide, and as mentioned above the reaction temperature is kept below -30 ° C. : Omura et al. J. Org. Chem. 41, 957 (1976).

The second method used to prepare intermediates to make useful antimicrobials is shown in the following table.

이다)를 산화시켜 4"-데옥시-4"-옥소-에리트로마이신 A 화합물로 전화시키는 산화반응에 관한 것이다. 이 반응에서는 산화제로서 N-클로로석신이미드와 몰메틸설파이드를 사용하여 실제에서, 이들 두 반응물은 처음에 0°C, 반응-불활성 용매에서 함께 혼합하고 10 내지 20분후 온도를 0 내지 -25°C로 내리고 알콜물질(Ⅰ') 또는 (Ⅱ')를 첨가하며 상기 언급한 온도를 유지시킨다. 2 내지 4시간 반응시킨후, 트리에틸아민 같은 3급아민을 첨가하고 반응 혼합물을 가수분해시킨후 끝처리한다.The second method is a 4 "-hydroxy substituent of the compounds of formulas (I ') and (II'), where Ac and R2 are each alkanoyl having 2 to 3 carbon atoms, R3 is hydrogen, and R2 and R₃ together

Is converted to a 4 "-deoxy-4" -oxo-erythromycin A compound. In this reaction, using N-chlorosuccinimide and molmethylsulfide as oxidants, in practice, these two reactants were first mixed together in a 0 ° C, reaction-inert solvent and after 10-20 minutes the temperature was 0--25 ° Lower to C and add alcohol (I ') or (II') and maintain the above mentioned temperature. After reacting for 2 to 4 hours, tertiary amines such as triethylamine are added and the reaction mixture is hydrolyzed and finished.

For the amount of reactants, one mole of N-chlorosuccinimide and one dimethylsulfide per mole of alcohol used is required. Experimentally, to complete the reaction quickly, it is preferred to use succinimide and sulfide reactant in an excess of 1 to 20 times and the tertiary amine used should correspond to the molar amount of succinimide used.

The reaction-inert solvent used in this reaction must be one that dissolves the reactants and does not react with the reactants or the formed product. Since the reaction is carried out at 0 to -25 ° C, it is preferable to use a solvent having a freezing point below the reaction temperature in addition to the above characteristics.

Such solvents or mixtures thereof include toluene, ethyl acetate, chloroform, methylene chloride or tetrahydrofuran. Solvents that meet the above requirements but have a freezing point above the reaction temperature can be used in small amounts by mixing with one or more of the preferred solvents. Particularly preferred solvent for this reaction is toluene-benzene.

Preferred compounds prepared by this reaction are 11,2'-diacetyl-4 "-deoxy-4" -oxo-ethromycin A 6,9-hemiketal, 2'-acetyl-4 "-deoxy-4 "-Oxo-erythromycin A 6,9-hemictal, 11,12-carbonate ester and 2'-acetyl-4" -deoxy-4 "-oxo-erythromycin A.

The reaction time is not limited and depends on the concentration, the reaction temperature and the intrinsic reactivity of the reactants.

If the reaction temperature is 0 to -25 ° C the reaction time is 2 to 4 hours.

In the order of addition of the reactants, as already mentioned, it is preferable to add the alcohol substance (I ') or (II') to the succinimide derivative and dimethyl sulfide mixed beforehand.

The two methods described above appear to be the same because of the selectivity of oxidation that occurs violently at the 4 ″ -hydroxy substituents, leaving other secondary alcohols in the unaffected molecule.

A 4 "-deoxy-4" -oxo compound of formula (II), useful as an intermediate, is prepared by treating a compound of formula (I) with alkanol anhydride (R2O) and pyridine.

In the above formula, R ₁ and R ₂ are each alkanoyl having 2 to 3 carbon atoms, R 3 is hydrogen, and Y is oxygen.

In practice, it is preferable to react ketone (I) with excess anhydride in a pyridine solvent and to use an excess of 4 times the anhydride in the reaction. The reaction is typically carried out at ambient temperature at which the reaction time is about 12 to 24 hours.

Removal of the alkanoyl sites at the 2'-position of the intermediates, ketones (I) (Y = 0) and (II), involved 2'-alkanoyl-4 "-deoxy-4" -oxo-erythromycin A The compound is carried out by solvolysis with excess methanol and stirring overnight at room temperature. Methanol is removed and, if necessary, the residue is subsequently purified to yield compounds of formula (I) (Y = 0) and (II), wherein R 'is hydrogen.

As already mentioned, the ketones of formulas (I) (Y = 0) and (II) are the 4 ″ -deoxy-4 ”-amino-erythromycin A of formulas (III) and (IV), which are the antimicrobial agents of the present invention. Useful intermediates for the preparation of the Preferred intermediates are 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin A 6, 9-hemictal 11, 12-carbonate ester and 4 "-deoxy-4" -oxo-erythromycin A 6, 9-hemichemal 11, 12-carbonate esters.

There are various synthetic methods for producing the antimicrobial agents of structural formulas (III) and (IV) from essential ketones (I) (Y = 0) and (II).

4 "-deoxy-4" -amino-erythromycin A of formula (III) is prepared by condensing the ketone of formula (II) with an ammonium salt of lower alkanoic acid and then reducing the imine produced in situ. "Lower alkanoic acid" means an acid having 2 to 4 carbon atoms.

In practice, a solution of ketone (II) dissolved in lower alkanol such as methanol or isopropane is treated with an ammonium salt of lower alkanoic acid such as acetic acid, and then the cooled reaction mixture is treated with sodium cyanoborohydride as a reducing agent. The reaction is allowed to proceed for several hours at room temperature, followed by hydrolysis and product separation.

One mole of ammonium alkanoate is required per mole of ketone, but it is preferred to use it in excess of 10 times to quickly and completely form imine. This overuse has a slightly detrimental effect on the quality of the product.

The amount of reducing agent used per mole of ketone is preferably about 2 moles of sodium cyanoborohydride per mole of ketone used.

The reaction temperature depends on the concentration, reaction time and intrinsic reactivity of the reactants and the reaction is almost complete after 2 to 3 hours at room temperature, which is the preferred reaction temperature.

When methanol is used as the lower alkanol solvent, the alkanoyl group is solvolyzed at the 2'-position. In order to prevent such a site from being removed, it is preferable to use isopropanol as a reaction solvent.

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

It is preferred to use the basic product of the final product to separate the desired 4 "-deoxy-4" -amino-erythromycin A derivative from the non-basic by-product or starting material. Thus, an aqueous solution of the product is extracted with increasing pH gradually so that the basic or non-basic material is extracted at a low pH and the product is extracted at a pH of 5 or more. The extraction solvent, ethyl acetate or diethyl ether is backwashed with brine and water, dehydrated over sodium sulfate and the solvent is removed to give the product. If desired, purification can be carried out by column chromatography on silica gel by known methods.

As already mentioned, the solvolysis of the 2'-alkanoyl group from the 2'-alkanoyl-4 "-deoxy-4" -amino-erythromycin A derivative prevents the methanol solution of the compound overnight at ambient temperature. It is effective to carry out.

이고 R₁이 탄소수 2 내지 3의 알카노일 또는 수소인 구조식(Ⅱ)케톤의 환원성아민화 반응동안 구조식(Ⅲ) 및 (Ⅳ) 화합물에 관련된 아민이 생성되며, 이를 반응식으로 표시하면 다음과 같다.R₂ and R₃ together

And amines related to the compounds of the formulas (III) and (IV) are produced during the reductive amination reaction of the formula (II) ketone, wherein R₁ is alkanoyl or hydrogen having 2 to 3 carbon atoms.

Amine products (III) and (IV) are usually separated by selective crystallization with diethyl ether.

Recrystallization of the mixture of formulas (III) and (IV) with acetone-water results in the formation of hemimetals in the amines of formula (IV) resulting in a single product of the compound of formula (III).

Direct synthesis of the amine compound of formula (IV) is the same route as described above, condensation of ketone of formula (I) with ammonium alkanoate and reduction of imine produced in the reaction system to sodium cyanoborohydride. .

Compounds of formula (IV), wherein R ', R3 and R', as described above, may be prepared by reducing the imine using hydrogen and a hydrogenation catalyst. Experimentally, ketone (I) is dissolved in lower alkanol, such as methanol or isopropane, treated with ammonium and a hydrogenation catalyst of lower alkanoic acid, such as acetic acid, and then shaken under hydrogen atmosphere until the reaction is complete.

One mole of ammonium alkanoate is required per mole of ketone, but it is preferred to use in excess of 10 times to form imine quickly and completely. This excessive use slightly harms the quality of the product.

There are many hydrogenation catalysts, but preferably 5 to 10 percent pd / c with Ranickel and the amount is determined by how quickly the reaction is completed and 10 to 200 percent of the weight of the compound of formula (I) is effective.

The pressure of the hydrogen gas in the hydrogenation vessel affects the reaction rate. In order to facilitate the reaction time, it is preferable to use an initial pressure of 50 psi, and the reduction reaction is preferably performed at ambient temperature.

The reaction time depends on the temperature, the pressure, the concentration of the reactants and the intrinsic reactivity of the reactants. Under the preferred conditions already mentioned, the reaction is completed within 12 to 24 hours.

The product is separated by filtration and removal of the catalyst under vacuum, the residue being treated successively with water and the basic product is extracted with water at the pH change described above to separate the product from the nonbasic material.

As already mentioned, when methanol is used as the lower alkanol solvent, most of the alkanoyl groups in the 2'-position are solvent decomposed. It is preferable to use isopropanol as the reaction solvent in order to avoid the removal of such sites.

-CH₃)로 전환시킨후 이를 환원시키는 것이다.A second synthetic process for preparing the antimicrobial 4 "-deoxy-4" -amino-erythromycin A of formula (IV) first involves the preparation of the ketone of formula (I) (Y = 0) with an oxime or oxime derivative (ie Y = N-). OH or NO

-CH₃) and then reduce it.

Oximes of ketone (I) (Y = 0) are prepared by reacting the ketone with hydroxylamine hydrochloride and barium carbonate in the presence of methanol or isopropanol. In practice, using an excess of hydroxylamine, i.e., 3 times excess, results in good yields for the desired intermediate. Ambient temperature and excess hydroxylamine are used to produce the desired oxime derivative during the reaction period of 1 to 3 hours. Barium carbonate is used in twice the amount of hydroxylamine hydrochloride used. The product is added to water and basified to pH 9.5 and extracted by extraction with a solvent that does not mix with water, ie ethyl acetate.

Alternatively, the reaction mixture is filtered, the filtrate is concentrated and dried in vacuo, and the residue is partitioned between water and a solvent which is not mixed with water at pH 9.0 to 9.5.

-CH₃)의 0-아세틸옥심 화합물은 상응하는 옥심을 아세틸화시켜 제조한다.Structural Formula (I) (Y = N-0 =

0-acetyloxime compound of -CH 3) is prepared by acetylating the corresponding oxime.

Experimentally, one mole of oxime is reacted with one mole of acetic anhydride in the presence of one mole of pyridine or triethylamine.

An excess of anhydride and pyridine, ie 30-40%, is preferred to facilitate completion of the reaction. The reaction is most preferably carried out overnight at room temperature in the presence of an aprotic solvent such as benzene or ethyl acetate. When the reaction is complete, water is added, the pH is adjusted to 9.0 and the product is separated from the solvent layer.

2'-acetyl-4 "-deoxy-4" -oxo-erythromycin A oxime, 2 'is preferred among the oxime and oxime derivatives which are useful intermediates for making 4 "-deoxy-4" -amino-erythromycin A antibacterial agents. -Acetyl-4 "-deoxy-4" -oxo-erythromycin A 0-acetyloxime, 4 "-deoxy-4" -oxo-erythromycin A oxime and 4 "-deoxy-4" -oxo-erythro Mycin A 0-acetyloxime.

-CH₃)의 환원반응은 옥심 또는 옥심유도체를 이소프로판올 같은 저급알칸올에 녹여 초기압력 1000psi의 수소대기하에 실온에서 철야 라니니겔 촉매로 촉매적 수소화시켜 수행한다. 소비된 촉매를 여과하고 여액으로부터 용매를 제거하여 원하는 구조식(Ⅳ)의 4"-데스옥시-4"-아미노 항균제를 수득한다. 만약 본반응에서 메탄올을 용매로 사용할 경우 2'-알카노일 부위가 가용매분해된다. 이 부반응을 제거하기 위해 이소프로판올을 사용한다.Ketone derivatives (Y = N-OH or NO-

Reduction of -CH₃) is carried out by dissolving an oxime or oxime derivative in a lower alkanol, such as isopropanol, and catalytic hydrogenation with a Rani Nigel catalyst overnight at room temperature under a hydrogen atmosphere at an initial pressure of 1000 psi. The spent catalyst is filtered and the solvent is removed from the filtrate to obtain the 4 "-desoxy-4" -amino antimicrobial agent of the desired structure (IV). If methanol is used as the solvent in this reaction, the 2'-alkanoyl moiety is solvolyzed. Isopropanol is used to eliminate this side reaction.

Preferred among the 4 "-deoxy-4" -amino-erythromycin A antibacterial agents of the formulas (III) and (IV) are 4 "-deoxy-4" -amino-erythromycin A 6, 9-hemichetal 11, 12 Epimer of -carbonate ester with 4 "-deoxy-4" -amino-erythromycin A, 4 "-deoxy-4" -amino-erythromycin A 11, 12-carbonate ester.

The chemotherapeutic action of the compounds of formulas (III) and (IV) of the present invention preferably uses pharmaceutically nontoxic salts of the compounds. Water-insoluble, highly toxic, or poorly crystalline salts are unsuitable or undesirable for use in scarce pharmaceutical formulations, but water-insoluble or toxic salts will degrade the salts and convert them into the corresponding pharmaceutically toxic bases as described above. Can be converted to the desired pharmaceutically toxic acid addition salt.

Acids that provide pharmaceutically harmless anions include hydrochloric acid, bromic acid, iodide, nitric acid, sulfuric acid or sulfurous acid, phosphoric acid, acetic acid, lactic acid, citric acid, succinic acid, maleic acid, gluconic acid and aspartic acid.

As already mentioned, the stereochemistry of the starting materials used to prepare the antimicrobial agents of the invention is that of natural materials. Oxidizing the 4 "-hydroxyl group to a ketone followed by 4" -amine conversion of the ketone means that the stereochemistry of the 4 "-substituent is changed from that of the natural product. Thus, Compound (I) (Y = Conversion of 0) and (II) to amines according to one of the methods described above results in the formation of two epimeric amines.

Experimentally it is observed that both epimeric amines are present in the final product in various ratios depending on the choice of synthesis method. If one epimer prevails in the isolated product, the epimer can be purified by recrystallization with a suitable solvent with a constant melting point. Other epimers present in small quantities in the original isolated solid material are the dominant product in the mother liquor. This can be recovered by known methods, for example, by evaporation of the mother liquor and the recrystallization of the residue repeatedly to give a product with a constant melting point.

The mixture of epimers can be separated by known methods, but for practical reasons it is preferred to separate the mixture from the reaction.

However, more preferably, the mixture of epimers is recrystallized one or more times with a suitable solvent, purified by column or high pressure liquid chromatography, solvent partitioning or treatment in a suitable solvent. The tablet, which does not require epimer separation, removes unnecessary materials such as starting materials and undesirable byproducts.

Absolute stereochemical studies of epimers are not yet complete. However, two epimers of a given compound exhibit the same activity, ie antimicrobial activity.

The above-mentioned 4 "-deoxy-4" -amino-erythromycin A derivatives are Gram-positive bacteria such as Staphylococcusaureus and Streptococcus Pyogenes, and Gram-like or spherical Gram. It shows the activity of living body against negative bacteria. This activity can be quickly determined by the testing of various organisms in brain-heart leaching media by 2-fold dilution technique.

By the activity of these living organisms, it can be seen that it can be used locally in the form of ointments, creams, sterilization of hospital room products and industrial bactericides such as water treatment, mucus control, paint and wood preservation.

For the use of a living body for topical use, the compound is mixed with a pharmaceutically toxic carrier (eg vegetable oil or mineral oil or softening cream. In a similar manner, the compound is a liquid carrier such as water, alcohol, glycol or mixtures thereof. Or dissolved or dispersed in a solvent or in another pharmaceutically inert medium (the medium should not have a deleterious effect on the active ingredient).

For this purpose generally 0.01 to 10% by weight of the active ingredient in the total composition weight is used.

In addition, many of the compounds of the present invention and these acid addition salts have been found to be gram positive and Gram negative bacteria (e.g. Pasteurella multocide) and Neisseria sicca These in vivo activities are more limited for sensitive organisms and are determined by conventional methods of infecting test organisms in mice of almost uniform weight and then treating the test compounds by oral or subcutaneous administration. In 10 mice are inoculated intraperitoneally in a diluted culture containing 1-10 times LD ₁₀₀ (minimum concentration required to kill 100% organic).

The comparative test was performed at low concentrations in the test mice to observe their toxicity. Test compounds are administered 0.5 hours after inoculation and 4, 24 and 48 hours after inoculation. The surviving mice are left for 4 days after the final treatment and the number of surviving mice is recorded.

When used in in vivo testing, the new compounds can be administered orally or parenterally (eg, subcutaneously or intramuscularly) at 1 mg / kg to 200 mg / kg per day.

The dosage is preferably 5 mg / kg to 100 mg / kg per day, and more preferably 5 mg / kg to 50 mg / kg per day for vehicles suitable for parenteral injection, water, isotonic saline, isotonic dextrose, rinser solution. Non-aqueous substances such as aqueous substances or plant fat oils (cotton seeds, peanut oil, corn, sesame seeds), dimethylsulfoxide and other non-aqueous bakes that are nontoxic to volume and width without compromising the therapeutic effect of the formulation (e.g. glycerol, Propylene glycol, sorbitol). In addition, it is desirable to make a composition suitable for solution preparations prior to administration. Such compositions include liquid diluents such as propylene glycol, diethyl carbonate, glycerol, sorbitol, and the like; Buffers, degrading enzymes, local anesthetics, and inorganic salts to achieve the desired pharmaceutical properties.

These compounds are mixed with pharmaceutically inert carriers, such as solid diluents, aqueous vehicles, and non-toxic organic solvents, to form capsules, tablets, lozenges, troches, anhydrous mixtures, suspensions, solvents, ariers and parenteral solutions. Or in suspension. In general, the compounds are used in various dosage forms at a concentration of 0.5 to 90% by weight of the total composition weight.

The following examples illustrate the invention in detail and thereby do not limit the scope of the invention.

Example 1

2'-acetyl-4 "-deoxy-4" -amino-erythromycin A

60 g of Ranickel washed with 14.0 g of 2'-acetyl-4 "-deoxy-4" -oxoerythromycin A 0-acetyloxime and isopropanol was dissolved in 400 ml of isopropanol and shaken at room temperature under a hydrogen atmosphere of 1000 psi of superatmospheric pressure. Let's do it. The catalyst is filtered off and the filtrate is concentrated to give a white foamy material.

The residue is dissolved with 400 ml of isopropanol and mixed with 50 g of Ranickel washed with fresh isopropanol and then hydrogenated overnight at room temperature under an initial hydrogen pressure of 1000 psi. The catalyst is filtered off and the filtrate is concentrated in vacuo and dried to give 8.1 g of the desired product.

Example 2

In the method of Example 1, 4 ''-amino-erythromycin A homologues described below were prepared using 0-acetyloxime as starting material:

Example 3

4 "-deoxy-4" -amino-erythromycin A

2.17 g of 2'-acetyl-4 "-deoxy-4" -amino-erythromycin A is dissolved in 50 ml of methanol and stirred overnight at room temperature. The solvent is removed under reduced pressure and the residual foam is treated with 50 ml of a mixture of chloroform and 50 ml of water. Adjust the pH of the aqueous layer to 9.5, separate the organic layer, treat the chloroform layer with fresh water and adjust the pH to 4.0. The pH of the acidic layer containing the product is gradually adjusted to 5, 6, 7, 8, and 9 by addition of a base and extracted with fresh chloroform at each pH. Extracts at pH 6 and 7 contain mostly product and are combined and treated with fresh water at pH 4. The aqueous layer is again adjusted to pH 5, 6 and 7 and extracted with fresh chloroform at each pH. The chloroform extract at pH 6 was dehydrated over sodium sulfate and concentrated to give 249 mg of product as epimer mixture.

NMR (δ, CDCI 3): 3. 30 (1H) s, 3. 26 (2H) s, 2. 30 (6H) s and 1. 46 (3H) s.

In a similar manner, 2'-propionyl-4 "-deoxy-4" -amino-erythromycin A is subjected to methanol solvent hydrolysis to yield 4 "-deoxy-4" -amino-erythromycin A.

Example 4

4 "-deoxy-4" -amino-erythromycin A

3.0 g of 4 "-deoxy-4" -oxo-erythromycin A is dissolved in 30 ml of methanol under nitrogen atmosphere and 3.16 g of anhydrous ammonium acetate is added to the stirred solution. After 5 minutes 188 mg sodium cyanoborohydride is added to the reaction mixture with 5 ml of methanol and stirred overnight at room temperature. Pour the light yellow solution into 300 ml of water and adjust the pH to 6.0. The aqueous layer is extracted with 125 ml of diethyl ether at pH 6,7,7.5,8.9 and 10.

The extracts at pH 8, 9 and 10 are combined and washed with 125 ml of fresh water. The separated aqueous layer was washed with ether (1 × 100 ml) at pH7, ethyl acetate (1 × 100 ml) at pH7, ether (1 × 100 ml) at pH7.5, ethyl acetate (1 × 100 ml) at pH7.5 and pH8,9 and Extract with ethyl acetate (1 × 100 ml) at 10. Ethyl acetate extracts at pH 9 and 10 were combined, washed with saturated brine solution, dehydrated over sodium sulfate and the solvent was removed under vacuum to give 30 mg of the desired product in the form of an ivory foam in the form of epimer mixture.

Example 5

4 "-deoxy-4" -amino-erythromycin A (single epimer)

10.0 g of an epimer mixture of 2'-acetyl-4 "-deoxy-4" -amino-erythromycin is dissolved in 150 ml of methanol and stirred for 72 hours at room temperature under nitrogen atmosphere. 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 tilted and 150 ml of fresh water is added, the pH of the aqueous layer is adjusted to 5 and the chloroform layer is separated. The pH of the aqueous layer is continuously adjusted to 5. 5, 6, 7, 8 and 9 and extracted with 100 ml of fresh chloroform, respectively. The chloroform extracts at pH 6, 7 and 8 were combined and washed sequentially with water and saturated brine solution, then dehydrated over sodium sulfate and the solvent removed under reduced pressure to give an epimer mixture of 4 "-deoxy-4" -amino-erythromycin A. Get 2.9 g. Treatment of 1.9 g of the sample of the mixture with diethyl ether crystallizes the insoluble foam.

The solid is filtered and dried to give 67 mg of a single epimer of 4 "-deoxy-4" -amino-erythromycin A having a melting point of 140 to 147 ° C.

Example 6

11-acetyl-4 "-deoxy-4" -amino-erythromycin A 6,9-hemiketal

4.4 g 11-acetyl-4 "-deoxy-4" -oxo-erythromycin A 6,9-hemiketal and 4.38 ammonium acetate dissolved in 75 ml of methanol in 305 mg of 85% sodium cyanobohydro Add a ride. After overnight stirring at room temperature, the reaction mixture is poured into 300 ml of water and 250 ml of chloroform are added. Adjust the pH of the aqueous layer to 9.8, separate the chloroform layer, extract the aqueous layer again with chloroform, combine the chloroform extracts, dehydrate on sodium sulfate and concentrate to give a white foamy material. Dissolve in the stirred mixture, adjust the pH to 4.9, separate the chloroform, decanter and adjust the pH of the aqueous layer to 5, 6, 7 and 8 and extract with fresh chloroform at each pH. Combine extracts at pH 6 and 7, wash with saturated brine solution, dehydrate on sodium sulfate and remove solvent to afford 1.72 g of the desired product as a white foam. The product is dissolved in a small amount of diethyl ether and then treated with hexane to make cloudy.

The crystalline product formed is filtered and dried to yield 1.33 g of product having a melting point of 204.5 to 206.5 ° C.

NMR (δ, CDCI 3): 3.31 (2H) s, 3.28 (1H) s, 2.31 (6H) s, 2.11 (3H) s and 1.5 (3H) s.

Example 7

In the method of Example 6, the following compounds were prepared using isopropanol instead of methanol as a reaction solvent and 4 " -deoxy-4 " -oxo-epitamycin A as starting materials.

Example 8

4 "-deoxy-4" -amino-erythromycin A 6,9-hechemal 11,12-carbonate ester

189 g- of 4 "-deoxy-4" -oxo-erythromycin A 6,9-hemiketal 11,12-carbonate ester is added to a solution dissolved in 1200 ml of methanol at room temperature with stirring of 193 g of ammonium acetate. After 5 minutes the resulting solution was cooled to -5 ° C and then treated with 13.4 g of 85% sodium cyanoborohydrogen in 200 ml of methanol for 45 minutes.

The cooling bath is removed and the reaction mixture is stirred overnight at room temperature.

The reaction mixture was reduced in volume to 800 ml in vacuo and added to a stirred mixture of 1800 ml of water and 900 ml of chloroform. 6N hydrochloric acid is added to adjust the pH to 6.2-4.3 and the chloroform layer is separated.

Chloroform is mixed with 1 L of water and the pH is adjusted to 9.5. The organic layer is separated, dehydrated over sodium sulfate and concentrated under reduced pressure to give 174 g of a white foamy material. The residue is dissolved in 1 L of water and 500 ml of ethyl acetate and the pH is adjusted to 5.5. The ethyl acetate layer was separated, and the aqueous layer was gradually adjusted to pH 5.7 and 9.5, and the pH was adjusted and extracted with 500 ml of fresh ethyl acetate. The ethyl acetate extract at pH 9.5 was dehydrated over sodium sulfate and concentrated in vacuo. Drying yields 130 g of product.

120 g of residual foamy material is dissolved in a mixture of 1 liter of water and 1 liter of methylene chloride and the pH of the aqueous layer is adjusted to 4.4, 4.9 and 9.4, respectively, and extracted with 1 liter of fresh methylene chloride at each pH. The ethylene chloride extract at pH 9.4 was dehydrated over sodium sulfate and concentrated under reduced pressure to give 32 g of a white foamy product which was crystallized from 230 ml of acetone-water (1: 1, V: V) to give 28.5 g of crystalline epimer.

NMR 100 Mz (δ, CDC1 2): 5.20 (1H) m, 3.37 (1.5H) s, 3.34 (1.5H) s, 2.36 (6H) s, 1.66 (3H) s and 1.41 (3H) s.

Example 9

Separation of Epimers of 4 "-Deoxy-4" -Amino-erythromycin A 6,9-hemical 11,12-carbonate ester

Filled with Gf 254 silica gel immersed in formamide and eluted with chloroform, 200 mg on a pressurized check chromatography column (1/2 "% 9 cm) at 4.7 cc / min under a pressure of 240 psi and a stroke size of 10 ml. Collect the fractions 14-21 and 24-36.

Fractions 14-21 are combined, concentrated to 50 ml, water (50 ml) is added and the pH adjusted to 9.0. The chloroform layer was separated, dehydrated over sodium sulfate and concentrated to give 106 mg of white foamy material, which was treated with diethyl ether to crystallize the foamy material. After stirring for 1 hour at room temperature, the crystalline product was filtered and dried to obtain 31.7 mg of the desired product having a melting point of 194 to 196 ° C.

NMR 100 Mz (δ, CDC1 3): 5.24 (1H) d, 5.00 (1H) t, 3.40 (3H) s, 2.40 (6H) s, 1.66 (3H) s and 1.40 (3H) s.

The fractions 24-36 were combined and proceeded as above to give 47.1 mg of the same white foamy material as in Example 14 described below.

Example 10

Stir 10.7 g of ammonium acetate in a suspension of 11.1 g of 2'-acetyl-4 "-deoxy-4" -oxoerythromycin A 6,9-hechemal 11,12-carbonate ester in 300 ml of isopropanol at room temperature. While adding.

After 5 minutes, 747 mg of sodium cyanoborohydride dissolved in 130 ml of isopropanol was added over 30 minutes, and the resulting reaction compound was stirred overnight at room temperature. Pour the light yellow solution into 1100 ml of water, add 400 ml of diethyl ether, adjust the pH to 4.5 and separate the ether layer. The aqueous layer is basified to pH9.5 and extracted twice with 500 ml of chloroform each time, the chloroform extracts are combined, dehydrated over sodium sulfate and concentrated to give 7.5 g of a yellow foam. The residue was recrystallized from diethyl ether to give 1.69 g.

The mother liquor is treated with 75 ml of water and the pH adjusted to 5.0, then the ether layer is replaced with 75 ml of fresh ether and the pH adjusted to 5.4. Replace the ether with ethyl acetate and raise the pH to 10. The basified aqueous layer is extracted twice with 75 ml of ethyl acetate each time, and the ethyl acetate extract is dehydrated over sodium sulfate, concentrated and dried. Residual foamy material (1.9 g) is added to a mixture of 75 ml of water and 50 ml of diethyl ether and the pH is adjusted to 5.05. The ether was separated and the aqueous layer was gradually pH 5.5 ,. Adjust to 6076.05 and 8.0 and extract with 50 ml of fresh diethyl ether.

The pH was finally adjusted to 9.7, the aqueous layer was extracted with 50 ml of ethyl acetate, the ether extract at pH 6.0 was mixed with 75 ml of water and the pH was adjusted to 9.7. The ether layer is separated, dried and concentrated in vacuo to yield 460 mg of white foamy material.

NMR 100Mz (δ, CDC1₃): 5.20 (1H) t, 3.43 (2H) s, 3.40 (1H) s, 2.38 (6H) s, 2.16 (3H) s, 2.70 (3H) s and 1.53 (3H)

NMR data show that the product is an epimer of 2'-acetyl-4 "-deoxy-4" -amino-erythromycin A 6,9-hemical 11,12-carbonate ester.

1.69 g of the product is 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 and the aqueous layer is extracted with fresh ether (75 ml) at pH 5.05 and 5.4 and ethyl acetate (2 x 75 ml) at pH 9.7. The combined ethyl acetate extracts were dehydrated over sodium sulfate and concentrated under reduced pressure to yield 1.26 g of white foamy material. This residue was crystallized to give 411 mg of product having a melting point of 193 ° to 196 ° C. (decomposition). The mother liquor is concentrated and dried, the residue is dissolved in hot ethyl acetate, the solution is left overnight at room temperature and the precipitated crystalline solid is filtered and dried to further give 182 mg of product having a melting point of 198 to 202 ° C. (decomposition). .

NMR 100Mz (δ, CDC1₃): 5.10 (1H) t, 3.34 (2H) s, 3.30 (1H) s, 2.30 (6H) s, 2.08 (3H) s, 1.62 (3H) s, 1.48 (3H) s.

NMR and data show that the product is an epimer of 2'-acetyl-4 "-deoxy-4" -amino-erythromycin A 11,12-carbonate ester.

In a similar manner, when the method of Example 10 is repeated, 2'-propionyl-4 "-deoxy-4" -oxo-erythromycin A 6,9-hemictal 11,12-carbonate ester is used as starting material. Using 2′-propionyl-4 ″ -deoxy-4 ″ -amino-erythromycin A 6,9-hemical 11,12-carbonate ester and 2′-propionyl-4 ″ -deoxy-4 ″- Amino-erythromycin A 11,12-carbonate ester is obtained.

Example 11

400 mg 2'-acetyl-4 "-deoxy-4" -amino-erythromycin A 6,9-hemictal-11,12-carbonate.

The ester is dissolved in 20 ml of methanol and stirred overnight at room temperature. Pour the reaction solution into 100 ml of water and add 50 ml of ethyl acetate. The pH is adjusted to 9.5, the organic layer is separated, and then repeatedly extracted with 50 ml of fresh ethyl acetate.

The combined ethyl acetate extracts were dehydrated over sodium sulfate and concentrated to give 392 mg of white foamy material, which was then treated with diethyl ether and crystallized by scraping a test tube with a glass rod, left at room temperature for 30 minutes, and then filtered and dried to obtain crystalline solids. The product is obtained and the mother liquor is retained. The NMR analysis shows that the product is the same as the material prepared in Example 13 described later.

NMR 100Mz (δ, CDC1₃): 3.26 (3H) s, 2,32 (6H) s, 1.61 (3H) s and 1.44 (3H) s

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

The retained mother liquor is concentrated in vacuo to give 244 mg of white foamy material. This product is the same as the product of Example 8.

The NMR data show that the product is an epimer mixture of 4 "-deoxy-4" -amino-erythromycin A 6,9-hechemal 11,12-carbonate esters, which is identical to the product of Example 8. have.

Example 12

In a similar manner as in Example 11, 2'-propionyl-4 "-deoxy-4" -amino-erythromycin A 6,9-hechemal 11,12-carbonate ester was subjected to methanol solvent addition decomposition to 4 "-de Obtain oxy-4 "-amino-erythromycin A 11,12-carbonate ester and 4'-deoxy-4" -amino-erythromycin A 6,9-hemiketal 11,12-carbonate ester.

Example 13

8 g of an epimer mixture of 4 "-deoxy-4" -amino-erythromycin A 11,12-carbonate ester obtained from the amorphous product of Example 8 was dissolved in 50 ml of diethyl ether and the product was scraped with a glass rod. Crystallize. After stirring for 20 minutes, the crystalline product was filtered and dried to yield 1.91 g of a product having a melting point of 198.5 ° to 200 ° C.

NMR 100MZ (δ, CDC1₃): 3.26 (3H) s, 2.30 (6H) s, 1.61 (3H) s 1.45 (3H) s

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

Example 14

1 g of the epimer of Example 13 is dissolved in 20 ml of acetone and heated in a steam bath until the boiling point is reached. Molar (25 ml) is added and the resulting solution is stirred at room temperature. After stirring for 1 hour, the formed precipitate was filtered and dried to give 581 mg of a product having a melting point of 147 to 149 ° C.

NMR 100 Mz (δ, CDC1 3): 5.12 (1H) d, 3.30 (3H) s, 2.30 (6H) s, 1.62 (3H) s and 1.36 (3H) s.

The NMR data shows that the product is a single epimer of 4 "-deoxy-4" -amino-erythromycin A 6,9-hemical 11,12-carbonate ester and is identical to the epimers of fractions 9 to 36 in Example 9. It can be seen.

Example 15

4 "-deoxy-4" -amino-erythromycin A

20 g of 4 "-deoxy-4" -oxo-erythromycin A, 31.6 g of ammonium acetate, and 10 g of 10% pd / c are dissolved in 200 ml of methanol and shaken overnight at ambient temperature under a hydrogen atmosphere at an initial pressure of 50 psi. The spent catalyst is filtered, concentrated in vacuo and dried. The residue is partitioned pH-5.5 to water-chloroform. The aqueous layer is separated, the pH is adjusted to 9.6 and chloroform is added. The organic layer is separated, dehydrated over sodium sulfate, concentrated and dried under reduced pressure. The remaining liquid colored foam (19 g) was treated with 150 ml of diethyl ether for 30 minutes at room temperature and the resulting solid was filtered and dried to yield 9.45 g of a single epimer indistinguishable from Example 5.

The diethyl ether filtrate is concentrated and dried to yield 6.89 g of a product consisting of other epimers containing some impurities.

Example 16

4 "-deoxy-4" -amino-erythromycin A

2 g of 4 "-deoxy-4" -oxo erythromycin A, 311 g of ammonium acetate and 2.0 g of rannickel are dissolved in 50 ml of methanol and shaken overnight at room temperature under a hydrogen atmosphere at an initial pressure of 50 psi. An additional 3.16 g of ammonium acetate and 2.0 g of Ranickel were added and hydrogenation continued for 5 hours.

The solid is filtered off and the filtrate is concentrated in vacuo and dried. The residue is added to the mixture of water-chloroform with stirring and the pH is adjusted to 6.4 to 5.5. The aqueous phase is separated, the pH adjusted to 9.6 and fresh chloroform added. The chloroform extract is separated, dehydrated over sodium sulfate and concentrated under reduced pressure to afford 1.02 g of yellow foamed product. The predominant isomer has the opposite arrangement at 17 compared to the compound of Example 5.

Example 17

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

4.66 g of anhydrous ammonium acetate was stirred in a solution of 4.5 g of 2'-acetyl-4 "-deoxy-4" -oxo-erythromycin B (see US Patent No. 3,884,903) in 45 ml of isopropanol under a nitrogen atmosphere. While adding. After 10 minutes, 376 mg sodium cyanoborohydride is added to the reaction mixture together with 10 ml of isopropanol and stirred overnight at room temperature. Pour the light yellow solution into 400 ml of water and adjust the pH to 6.0. The aqueous layer is extracted at pH 6,7,7,5,8,9 and 10 with 250 ml of diethyl ether and the extracts at pH 8,9 and 10 are combined and washed with 250 ml of fresh water. The separated aqueous layer was washed with ether (1% 100 ml) at pH 7, ethyl acetate (1 x 100 ml) at pH 7, ether (1 x 100 ml) at pH 7.5 ethyl acetate (1 x 100 ml) at pH 7.5 and pH 8,9 And extracted with ethyl acetate (1 × 100 ml) at 10, respectively. The ethyl acetate extracts at pH 9 and 10 are combined, washed with saturated brine solution, dehydrated over sodium sulfate and the solvent is removed in vacuo to yield an epimer mixture of the desired product on a creamy colored product.

In a similar manner, 4 "-deoxy-4" -amino-erythromycin B is prepared from -4 "-deoxy-4" -oxo-erythromycin B.

Example 18

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

4.34 g of 2'-acetyl-4 "-deoxy-4" -amino-erythromycin B is dissolved in 100 ml of methanol and stirred overnight at room temperature. The solvent is removed under reduced pressure and the residual foam is treated with a mixture of 100 ml of chloroform and 100 ml of water. Adjust the pH of the aqueous layer to 9.5, separate the organic layer, treat the chloroform layer with fresh water and adjust the pH to 4.0. The pH of the acidic layer containing the product is gradually adjusted to 5,6,7,8 and 9 by addition of base and extracted with fresh chloroform at each pH. Extracts at pH 6 and 7 contain most of the product and are combined and treated with fresh water at pH 4. The aqueous layer is again adjusted to pH 5, 6 and 7 and extracted with fresh chloroform at each pH. The chloroform extract at pH 6 is dehydrated over sodium sulfate and concentrated to give the product as an epimer mixture.

Claims (1)

In reducing compounds of the following formulas (I) and (II), when Y "is NH and Y 'is NH, N-OH or N-OCOCH₃, reduction is carried out by hydrogenation in the presence of a catalyst, and Y 'Or Y' is NH, and in the case of compounds which are naturally produced by condensing the corresponding ketones of formulas (I) and (II) with ammonium of alkanoic acid, the reduction reaction is carried out in the presence of a hydride, Next, the process for preparing 4 "-amino-epimeric compounds of the general formulas (III) and (IV).

In the above general formula R 'and R' are each hydrogen or alkanoyl of 2 to 3 carbon atoms; R2 is alkanoyl of 2 to 3 carbon atoms; R 3 is hydrogen; R₂ and R₃ together

ego; R₃ and R₄ together

Is; Y 'is NH, N-OH or N-OCOCH₃; Y "is NH.