FI68404C - FREQUENCY REQUIREMENT FOR THERAPEUTIC USE OF THERAPEUTIC 4 "DEOXI-4" -AMINOERYTROMYCIN-A DERIVATIVES - Google Patents

Description

68404

Process for the preparation of therapeutically useful 4 "-deoxy-4" -aminoerythromycin A derivatives

Separated by division of the application 780 354.

The present invention relates to a process for the preparation of new antibacterial agents. In particular, the invention relates to a process for the preparation of therapeutically useful 4 "-deoxy-4" aminoerythromycin A-10 derivatives of the formula

Ri J N (CiL) HO? Oi 32 O S * CtCH3> 2 R o 9¾ y \ rx 15 R2 °> //, ✓? ^ r '° R4 O ltt> 1 X. or T H0 1 R3 ° nS, jC. ^ Y r ^>

/ '0CH 0 A NH

3 X OCIL 2 20 111 IV 3 wherein R 1 and R 2 represent a hydrogen atom or an alkanoyl group having 2 to 3 carbon atoms is an alkanoyl group having 2 to 3 carbon atoms, R 1 is a hydrogen atom or 3 R 2 and R 2 together form a group E 1 or R 2a R 2 25 together form n, as well as their pharmaceutically

-O

to prepare acceptable acid addition salts.

Erythromycin is an antibiotic formed by culturing a strain of Streptomyces erythreus in a suitable culture medium (U.S. Patent No. 2,653,899). Erythromycin is composed of two species, A and B, which can be represented by Formula 2.

B

68404 v N (CU,) _ O v | 4 '3 2 0 .3 ^ 7 ^ / N5'. HO „S0 5 jllHO j R - ^ O 5" * Ί? Ρ 31τ ° · '. ^ Ν t> I4' lg 2 'y ^: i’oh ’’ OCH3 10

Erythromycin R A “5h

B -H

15 It is apparent from the structural formula that the antibiotic consists of three main components, the sugar moiety cladinose, the second sugar moiety containing a basic amino substituent called desosamine, and a 14-membered lactone ring called erythronolide A or B or the following macrolide ring. The numbering of the macrolide ring is without dots, one dot is used in the desosamine part and two dots in the cladinose part.

Numerous derivatives of erythromycin have been prepared in an effort to modify its biological or far-25 macodynamic properties.

U.S. Patent 3,417,077 describes the reaction product of erythromycin and ethylene carbonate as a highly active antibacterial agent. U.S. Patent No. 3,884,903 discloses 4 "-deoxy-4" -oxoerythromycin A and B derivatives as useful anti-30s.

Erythromycylamine, a 9-amino derivative of erythromycin A, has been the subject of extensive research / English Patent 1,100,505, Tetrahedron 35 Letters, 1645 (1967) and Croatica Chemica Acta, 39, 273 (1967 //), and its structure is somewhat contradictory! - 3 68404 to Tetrahydron Letters, 157 (1970) and English Patent 1 341 022 / U.S. Patent No. 3,983,103 discloses that erythromycylamine sulfonamide derivatives are useful as anti-bacterial agents. effect in vitro and in vivo ^ Ryden, et al., J. Med Chem., 16 1959 (1973) and Massey, et al., J. Med. Chem., 17, 105 (1974).

It has now been found that certain novel 4 "-deoxy-4" -10 aminoerythromycin A derivatives are excellent antibacterial agents.

Ri <; N (Cl 1) - λαΥιΙ! R ~ O.-L_ and T H0 1 'yX AXx 20 X CCH, ό X 1 NH_ m IV «, 3 and pharmaceutically acceptable acid addition salts thereof, wherein R 1 and R 2 are each hydrogen or 2- Alkanoyl having 3 carbon atoms, R 1 is alkanohydrogen having 2 to 3 carbon atoms, or ≤ 2 and 0 nm together are -C- and R 1 and R 2 together are -C-.

A preferred group of compounds 30 for these chemotherapists are the compounds of formula III. Particularly preferred compounds of this group are those in which O 11 R 2 and R 2 together form a group -C-.

4 68404

Another preferred group of antibacterial agents are compounds of formula IV. Of these, those having hydrogen and those in which R 1 and R 2 together form a group 5 O are particularly preferred.

II

C-.

Another object of the present invention is to provide intermediates from which antibacterial agents of formula III and IV can be prepared. These intermediates 10 have the formulas R, Λ nIchJ, 0? 0 Ϊ 3 2. \ NtCH)

Vs _ vS "" 'NCi 15 HO ι., Λ0Λ «^^“ Λ Ax ’*" Ά Av

s Xt *, 1 τ 'CX

/ 'OCH * 0 S "' 0

3 'OCIL

Wherein R 1 is hydrogen or alkanoyl having 2 to 3 carbon atoms, R 2 is alkanoyl having 2 to 3 carbon atoms,

O

Y is N-OH or N-O-CCH 2 R 2 is water or R 2 and R 2 may be

O

M

together be -C-.

Preferred among these intermediates are compounds of formula I, of which intermediates in which R 1 is hydrogen or acetyl are particularly preferred.

Another preferred group of intermediates are compounds of formula II, of which R 1 is acetyl are particularly preferred.

The invention also relates to a process for the preparation of intermediates of formula I and 35 of formula II 5 68404 Λ \ N (CH_), = O I 3 2 „Ac N (CH) _

vvri% s.VV

5 <»j> Ηο'γ '^' ^ '" 2 ° Y 0'γ-Ο ^' Ο ^ ΗαΛ fC ^ y la V-k ri.o 5 ^ 'V <£.

3 'OCH- I II 3 10 wherein Ac and are each alkanoyl having 2-3 carbon atoms, R. is hydrogen or R and R- o

II

15 may together be -C-, which method is characterized in that the formulas, Λ \ N (CH) = O I .Ac N (CHJ.

20 ° yS π "vy \ X

no ,,,, -) v sf ° \ X0X

"-K & A, 33 κ, ο-Ι Λ

XX CC K&X

The compounds of the reaction are reacted in a reaction-inert solvent at about -30 to -65 ° C with 1 mole of dimethyl sulfoxide and 1 mole of trifluoroacetic anhydride, followed by contacting the reaction mixture with at least 1 mole. with a mole of triethylamine.

In a preferred embodiment of this process, the compound of formula 1 'and II' is oxidized in a reaction-inert solvent which is methylene chloride.

6 68404

In another method according to the invention, ÖZW of the formulas Λ \ N (CHJ_ 5 = 0 I 3 2 .Ac N (CH_), γνη ”sA vy H0" "f ho '« 2 ° v' u / ¾

T 3 'CC1L

Compounds of formula I II 3 in which Ac and R2 are each alkanoyl having 2 to 3 carbon atoms, R is

D O

It is hydrogen, or and R 2 together can be -C-, which process is characterized in that the formulas Λ \ N (CU) 20 Ξ O I 3 2 .Ac N (CH-),

° yS ^ ‘VN \ A

J “° λ la ** Y yVmY

P? 1 R3 ° ~ 7> | The compounds of I · 3 II '' 5 are reacted in a reaction inert solvent at about 0 to -25 ° C with 1 mole of N-chlorosuccinimide and 1 mole of dimethyl sulfide, after which the reaction mixture is contacted with at least 1 mole of triethylamine. .

7 68404

In the process according to the invention, toluene or benzene is preferably used as the reaction-inert solvent.

Throughout the invention, the stereochemical designation of the sugar and macrolide ring 5 substituents is the same as for erythromycin A, with the exception of the possible epimerization of the 4 "position, using the designations of naturally occurring erythromycin A.

The processes for the preparation of antibacterial agents derived from 4 "-deoxy-4" -amino-10 erythromycin A according to the invention can be described by the following reaction schemes starting from 2'-alkanoyl erythromycin A or a derivative thereof:

Ac Ac = \> Γ "32 Γ No Γ" 3'2 ° <yY. ' n ογΥ 'n

20 »0„ „> _v /’C.O’SjA

| HO I '110 1 ho__I X L · O] c ΓΏ - ,, / γ

S ϊχ ^ ΟΗ Y

^ 'OCH X' OCH

25 1 'I (Y = 0) and 8 68404 Η ° ^ Α ^ X "3'2 HO γ ÄC \ rM3'2" 2 ° y r2 °

r.o —L A -> D F I

5 rl ίγγ 3 Λ γ> Υ

<>> roH It '' lX

X OCH 0 YT0

.3 x OCH

II 'II 3 10

As a first part of the process, oxidation of compounds 1 'and II' (Y = O) by reacting compounds I * and II1 with trifluoroacetic anhydride and dimethyl sulfoxide and then adding a tertiary amine such as triethylamine.

In practice, the reaction is carried out by introducing trifluoroacetic anhydride and dimethyl sulfoxide into a reaction-inert solvent at about -65 ° C. After about 10 to 15 minutes, the alcohol 1 'and II' are added at such a rate that the temperature remains at about -65 ° C and does not rise above -30 ° C. At temperatures above -30 ° C, the trifluoroacetic anhydride-dimethyl sulfoxide complex is not stable. The temperature is maintained at -30 to 65 ° C for about 15 minutes and then lowered to -70 ° C. The tertiary amine is added in one portion and the reaction mixture is allowed to warm for 10-15 minutes. The reaction mixture is then treated with water and the reaction product is isolated.

Reagents, trifluoroacetic anhydride and dimethyl sulfoxide are each required in 1 mole per 30 moles of starting alcohol. In practice, it has been found that it is preferable to use a 1-5-fold excess of anhydride and dimethyl sulfoxide to accelerate the reaction.

9 68404

The amount of tertiary amine should correspond to the amount of trifluoroacetic anhydride used.

The reaction-inert solvent used in this process should be one that dissolves the reactants sufficiently well and does not react more substantially with the reactants or reaction products. Since this oxidation is carried out at -30 to -65 ° C, it is preferable that, in addition to the above-mentioned properties, the solidification point of the solvent is below the reaction temperature.

Solvents or solvent mixtures that meet these requirements include toluene, methylene chloride, ethyl acetate, chloroform and tetrahydrofuran. Solvents which otherwise meet the above requirements, but which have a pour point above the reaction temperature, may be used in combination with preferred solvents in minor amounts. A particularly preferred solvent for use in the process is methylene chloride.

Preferred compounds of this method are 2'-acetyl-4 "-oxoerythromycin-A, 11,2'-deoxy-4" -oxoerythromycin-A-6,9-hemiketal and 2'-acetyl-4 "-deoxy -4 "-oxo-erythromycin A 6,9-hemiketaa-ni-11,12-carbonate ester.

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

In the most preferred order of reagent addition, the trifluoroacetic anhydride is first combined with dimethyl sulfoxide, and then the required alcohol compound is added. Keeping the temperature below -30 ° C proposed herein also follows Omura1 et al., J. Org. Chem., 41, 957 (1976).

Another process according to the invention for the preparation of intermediates from which useful antibacterial agents are obtained can be represented by the following reaction formulas: 68404? \ TVa? \ «· <Α.3» 2. -OCw

no — L ho 1 JL

V & V &, ^ "3 OCH3 10 '1 (γ = 0) and 15 H ° XAC <0 HO - N <CH.

20 picks up NcS VS

IU> j I _> 2 "Y γοΥ, Λ /> 1 Γο ,, / 0 ^ / R3 ° —jL Λ 25 * Λ '™ Κλ Λ 25' OCH, 0> C0 II '3 OCH, 11 II 3 This the second method comprises an oxidation reaction in which the compound 1 'or II' in which Ac and R_ are alkanoyl groups having 2 to 3 carbon atoms is hydrogen, or

O

n R 2 and R 2 are together a group -C, 4 "-hydroxy substituent is oxidized to give 4" -deoxy-4 "-oxoerythromycin -A compound.

35 11 68404

The process uses N-chloro-succinimide and dimethyl sulfide as oxidizing agents. In practice, the two reagents are first combined in a reaction-inert solvent at about 0 ° C. After 10 to 20 minutes, the temperature drops to 0 to 25 ° C, then the alcohol compound 1 'or II' is added, and the temperature is kept the same. After a reaction time of 2 to 4 hours, a tertiary amine such as triethylamine is added to the reaction mixture, the reaction mixture is hydrolyzed, and the desired product is recovered.

One mole of N-chlorosuccinimide and 1 mole of dimethyl sulfide per mole of alcohol compound are required for the reagents. Experiments have shown that it is advantageous to use a 1 to 20-fold excess of succinimide and sulfide reagents to speed up the reaction. The amount of tertiary amine used should correspond to the molar amount of succinimide used.

The reaction-inert solvent used in this process of the invention should be one that dissolves the reactants sufficiently well and does not react more substantially with the reactants or reaction products. Since the reaction is carried out at about 0 to -25 ° C, it is preferable that, in addition to the above-mentioned properties, the solidification point of the solvent is below the reaction temperature. Solvents or solvent mixtures meeting these requirements include toluene, ethyl acetate, chloroform, methylene chloride and tetrahydrofuran. Solvents which otherwise meet the above requirements but which have a solidification point above the reaction temperature may also be used in small amounts in combination with one or more preferred solvents. A particularly preferred solvent for use in this process is toluene-benzene.

Preferred compounds prepared by this method are 11,2'-diacetyl-4H-deoxy-4 "-oxoerythromycin 35 -A-6,9-hemiketal, 2'-acetyl-4" -deoxy-4 "-oxoerythromycin-A -6,9-hemiketal-11,12-carbonate ester and 2'-12,680,404 acetyl-4 "-deoxy-4 '' - oxoerythromycin A.

The reaction time is not critical and depends on the concentrations of the reagents, the reaction temperature and the reactivity of the reagents. At reaction temperatures of 50 ° C to -25 ° C, the reaction time is about 2 to 4 hours.

As mentioned above, the addition of the reagents is preferably carried out by adding the alcohol compound 1 'or II' to a premixed mixture of the succinimide derivative and dimethyl sulfide.

Both of the methods described herein can be considered surprising because they selectively oxidize only when the 4 "hydroxy substituent is oxidized, leaving other secondary alcohols in the molecule unchanged.

15 4 "-deoxy-4" -oxo intermediates of formula% ^ * 1χΛα, 3) 2

20 X

Wherein R 1 and R 2 are each alkanoyl having 2-3 carbon atoms and R 2 is hydrogen, is prepared by treating the formula R 1 N (CIL)., 3 ° O JL \ X 3 2

‘’ .Χ XtXX

Γ H0 1 no_L Js.

p. -, x ° Y

'v' X

'' OCH I J

68404 13 wherein Y is O is alkanoyl having 2 to 3 carbon atoms, alkanoic anhydride (R 2 O) and pyridine.

In practice, ketone I is contacted with anhydride used in excess of 5 in a pyridine solvent. The reaction preferably uses up to a 4-fold excess of anhydride.

The reaction is preferably carried out at room temperature. In this case, the reaction time is about 12 to 24 hours.

Removal of the alkanoyl group from the 2 'position of intermediate ketone I (Y = 0) or II is performed in a solvolysis reaction in which the 2'-alkanoyl-4 "-deoxy-4" -oxoertyromycin A derivative is stirred with excess methanol overnight at room temperature. Removal of methanol 15 and purification of the residual product, if necessary, gives compounds of formulas I (Y = O) and II in which R 1 is hydrogen.

As mentioned, the compounds of formulas I (Y = O) and II are intermediates to give the 4 "-deoxy-4" -aminoerythromycin A antibacterial agents of formulas III and IV of the present invention. Preferred intermediates in this class of compounds are 2'-acetyl-4 "-deoxy-4" -oxoerythromycin A-6,9-hemiketal-11,12-carbonate ester and 4 "-deoxy-4" -oxoerythromycin-25A-6,9 -hemiketaali-II, 12-carbonate.

Several different synthetic routes can be used to prepare antibacterial agents of formulas III and IV from ketones I (Y = O) and II.

The 4 "-deoxy-4" -aminoerythromycin-30-A compounds of formula III are prepared by condensing a ketone of formula II with an ammonium salt of a lower alkanoic acid and then reducing the imine formed in situ. The term "lower alkanoic acid" as used herein means an acid having 2 to 4 carbon atoms.

In practice, a solution of ketone II in a lower alkanol, such as methanol or isopropanol, is treated with ammonium salt of a lower alkanoic acid, such as acetic acid, and the cooled reaction solution is treated with a reducing agent, such as sodium cyanoborohydride. The reaction is carried out at room temperature for several hours, after which the reaction mixture is hydrolyzed and the product is isolated.

Although one mole of ammonium alkanoate is required per mole of ketone, it is preferable to use this in excess, up to a 10-fold excess, in order to complete and rapidly form the imine. Such an excess does not appear to have much detrimental effect on product quality.

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

The reaction time varies depending on the concentrations of the reagents, the reactivity and the reaction temperature. At the preferred reaction temperature, room temperature, the reaction is substantially complete after 2-3 hours.

When the lower alkanol solvent is methanol, as mentioned above, solvolysis of the alkanoyl group in the 2 'position takes place. To prevent the removal of this group, isopropanol is preferably used as a solvent in the reaction.

The preferred ammonium alkanoate in this reaction is ammonium acetate.

To isolate the 4 "-deoxy-4" -aminoerythromycin A derivative of the desired final product from non-basic by-products or starting materials, the basic nature of the final product is utilized. In this case, the aqueous solution of the product is gradually extracted at an elevated pH, whereby neutral or non-basic substances are extracted at lower pH ranges and the reaction product at a pH above 5. Extraction solvents, ethyl acetate or diethyl ether, are extracted with brine and water, dried over sodium sulfate, product 15 68404 is obtained by removing the solvent. Any further purification is carried out by known methods by column chromatography on silica gel.

As already mentioned above, solvolysis of the 2'-alkanoyl group 5 from the 2 "-alkanoyl-4" -deoxy-4 "-aminoerythromycin A derivative can be performed by allowing a methanolic solution of the compound to stand at room temperature overnight.

In the reductive amination of a ketone of formula II wherein and together form

10 O

The group -C-, and R 1 is alkanoyl or hydrogen having 2 to 3 carbon atoms, amines of both formula III and IV are formed. This can be represented by the following reaction scheme: 15 - R, H (CH3> 2 »» Jv VA-i „Χ ° Χ.ΛΑ o. ,,, / (^ NH.OCOCH, I 1 4 3 N, 20 CK * NaCNBH3 II ^ ° ah o 25 (R2 + R3 = -C-) (ΐ, α, 3> 2, N, CH)

- r ° 1X vN VX

30 0 = <oi X „o-P ...... f H </ T

• f ^ CL, VO.

OCH3 / "ocni

3S III IV

O O

{R2 + R3 = ~ C_) (R3 + R4 = 68404 16

The amines III and IV obtained can be conveniently separated from the diethyl ether by selective crystallization. Upon crystallization of a mixture of III and IV from an acetone-water mixture, the amine of formula IV forms a hemiketal from which compound III can be isolated as the sole product.

The direct synthesis of the amine compound of the formula IV is carried out as already discussed above by condensing ketone I with ammonium alkanoate and reducing the imine obtained in situ with sodium cyanoborohydride.

Compounds of formula IV in which and R 1 are as defined above are also prepared by reduction of the above imine with hydrogen and a suitable hydrogenation catalyst. The process is carried out by treating the appropriate ketone I in a lower alkanol such as methanol, an ammonium salt of a lower alkanoic acid such as ammonium acetate, adding a hydrogenation catalyst and shaking the reaction mixture under a hydrogen atmosphere until the reaction is substantially complete.

1 mole of ammonium alkanoate is required per mole of ketone, however, an excess, preferably up to a 10-fold excess, is preferably used in order for complete and rapid imine formation. Such an excess does not appear to have a major effect on shark quality.

Many different catalysts can be used as the hydrogenation catalyst, however, Raney nickel and 5 to 10% palladium carbon are preferred. They can be used in varying amounts depending on how quickly the no-30 reaction is desired to take place. Preferably 10 to 200% by weight of the amount of compound I is used.

The pressure of the hydrogen gas in the hydrogenation vessel also affects the reaction rate. To shorten the reaction time, it is preferable to use an initial pressure of 0.34 MPa. The most suitable temperature for carrying out the reduction is room temperature.

17 68404

The reaction time depends on several factors, such as temperature, pressure, reagent concentrations, and reactivity. Under the above-mentioned preferred reaction conditions, the reaction time is 12 to 24 hours.

5 The product is isolated by filtration of the catalyst and evaporation of the solvent in vacuo. The evaporation residue is then treated with water, and the product is isolated from non-basic substances by extracting the basic product from water at varying pH ranges, as described above.

As mentioned above, when methanol is used as the lower alkanol solvent, solvolysis of the alkanoyl group in the 2 'position occurs. To prevent the removal of this group, it is preferable to use isopropanol as the solvent in the reaction.

In another synthetic route, the 4 "-deoxy-4" -aminoerythromycin A-antibacterial agents of formula IV are prepared by first reacting ketone I (Y = O) to the oxime or oxime derivative, i.

0

II

Y = N-OH, or N-O-CCH 2, and then the oxime or oxime derivative is reduced.

The oxime of ketone I (Y = O) is prepared by reacting the ketone with hydroxylamine hydrochloride and barium sulfate in methanol or isopropanol at room temperature. In practice, an excess of hydroxylamine is used, up to a 3-fold excess, whereby the desired intermediate is obtained in good yield. By carrying out the reaction at room temperature and using an excess of hydroxylamine, the desired oxime-30 derivative is obtained in a reaction time of 1 to 3 hours. Barium carbonate is used in twice the molar amount of hydroxylamine hydrochloride used. The product is isolated by adding water to the reaction mixture, adjusting the pH to 9.5, and extracting with a water-immiscible solvent such as ethyl acetate.

68404 18

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

An O-acetyloxime compound of formula I.

O

II

(Y = N-O-CCH 2) is prepared by acetylation of the corresponding oxime. In practice, this is done by reacting one mole of oxime with one mole of acetic anhydride in the presence of one mole of pyridine or triethylamine. By using an anhydride and an excess of pyridine, preferably an excess of 30 to 40%, the reaction proceeds more completely. Preferably, the reaction is performed in an aprotic solvent such as benzene or ethyl acetate at room temperature overnight. After completion of the reaction, water is added to the mixture, the pH is adjusted to 9.0, and the product is isolated from the solvent layer.

Preferred oximes and oxime derivatives which are suitable intermediates in the preparation of 4 "-deoxy-4" -aminoerythro-20 mycin A antibacterial agents are 21-acetyl-4 "-oxoerythromycin A-oxime, 2'-acetyl-4" - deoxy-4 "-oxoerythromycin-AO-acetyloxime, 4" -deoxy-4 "-oxoerythromycin-A-oxime and 4" -deoxy-4 "oxoerythromycin-AO-acetyloxime.

II

The reduction of ketone derivatives (Y = N-OH or N-O-CCH 2) is carried out by catalytic hydrogenation by shaking a solution of the oxime or its derivative in a lower alkanol such as isopropanol in the presence of a Raney nickel catalyst under an atmosphere of initial pressure at 6.9 MPa overnight at room temperature. The catalyst is then filtered, and the solvent is removed from the filtrate to give the desired 4 "-deoxy-4" -amino antibacterial compound of formula IV. If methanol is used as the solvent in this reduction, it may undergo solvolysis of the 2'-alkanoyl group. To avoid this side reaction, isopropanol is used as the solvent.

68404

Preferred 4 "-deoxy-4" -aminoerythromycin A derivatives of formulas III and IV are 4 "-deoxy-4" -aminoerythromycin A-6,9-hemiketal-11,12-carbonate ester, 4 "-deoxy Both epimers of -4 "-aminoerythromycin-A and 4" -deoxy-5 4 "-aminoerythromycin-A-11,12-carbonate ester.

Of course, in utilizing the chemotherapeutic activity of the compounds of formulas III and IV of the present invention which form salts, it is preferred to use pharmaceutically acceptable salts. Although certain salts may be unsuitable or less suitable for such pharmaceutical use due to their water insolubility, high toxicity, or lack of crystalline form, water-insoluble or toxic salts may be converted to the corresponding pharmaceutically acceptable bases by cleavage, or alternatively may be converted to the desired drug. .

Acids whose anions are pharmaceutically acceptable include, for example, hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, sulfuric, phosphoric, acetic, lactic, citric, tartaric, succinic, maleic, gluconic and and aspartic acid.

As mentioned above, the stereochemistry of the starting materials of the antibacterial agents of the present invention is found in natural materials. By oxidizing the 4 "hydroxyl group to a ketone and then converting this ketone to a 4" amine, it is possible to change the stereochemistry of the 4 "substituent to a different product from the natural product. Thus, by converting compound I (Y = O) or II to an amine in one of the above ways Experiments have shown that both amine epimers are present in different proportions in the final product, depending on the chosen synthetic method 35. If the isolated product contains mainly one epimer, this epimer can be purified from the constant melting point by repeated crystallizations. , the minor amount of epimer in the original isolated product is the major epimer in mother liquor 5. It can be recovered by methods known in the art, for example, by evaporating the mother liquor and performing repeated recrystallizations of the residue until the melting point is constant.

Although the separation of these epimer mixtures by methods known in the art can be performed, for practical reasons these mixtures are preferably used as they are isolated. However, it is often advantageous to purify the epimer mixture at least once by crystallization from a suitable solvent, by purification by column or high pressure liquid chromatography, solvent partitioning or trituration in a suitable solvent. Such purification, while not necessarily separating the epimers, removes excess materials such as starting materials and unwanted by-products.

Absolute stereochemical determination of the epimers was not performed. However, both epimers of a particular compound have the same type of activity, e.g., antibacterial activity.

Novel 4 "-deoxy-4" -aminoerythromycin A-derivative women have in vitro activity against a variety of fram-positive microorganisms, such as Staphylococcus aureus and Streptococcus pyogenes, and certain gram-negative microorganisms. such as spherical or elliptical (cooks) 30 microorganisms. Their activity has been demonstrated in in vitro experiments in brain-heart infusion medium against various microorganisms using the standard double serial dilution technique. Due to their in vitro activity, they are useful for topical use as ointments, creams, etc., for sterilization purposes, e.g., sterilization of patient room equipment, and as industrial antimicrobes, e.g., for water treatment, slime control, paint and wood preservation.

For in vitro use, e.g. topical application, it is often most convenient to incorporate the selected compound into a pharmaceutically acceptable carrier, such as a vegetable or mineral oil or ointment base. They may also be dissolved or dispersed in liquid carriers or solvents such as water, alcohol, glycols or mixtures thereof, or other pharmaceutically inert women, i. media which do not have an adverse effect on the active substance. In such preparations, the active ingredient is generally used in an amount of from about 0.01% to about 10% by weight of the total composition.

In addition, several of the compounds of this invention and their acid addition salts are active against fram-positive and certain gram-negative microorganisms, e.g. Pasteurella multocida and Neisseria sicca, when administered orally and / or parenterally in vivo to animals and humans. . Their in vivo activity against harmful microorganisms is limited and can be determined by a conventional method by infecting mice of substantially equal weight with the test organism and then treating them orally or with a subcutaneous test compound. The experiment is performed by infecting 25, e.g., 10 mice intraperitoneally with an appropriately diluted culture containing about 1 to 10 times the dose of LD10Q (LD1Q0 is the lowest concentration of microorganism that causes 100% death). Control experiments are performed in Sima, in which 30 mice are given lower concentrations of infectious agent to detect possible variations in the viability of the test organism. The test compound is administered 0.5 hours after infection, and the administration is repeated after 24 and 48 hours. Surviving mice are monitored for four days 35 after the last treatment, and the number of survivors is recorded.

68404 22

When used in vivo, these novel compounds may be administered orally or parenterally, e.g., by subcutaneous or intramuscular injection, at doses of from about 1 to about 200 mg / kg of body weight per day. The preferred dosage is from about 5 to about 100 mg / kg body weight in color, and most preferably from about 5 to about 50 mg / kg.

For parenteral administration, the injection medium may be either an aqueous medium such as water, isotonic saline, isotonic dextrose-10 solution, Ringer's solution, or an anhydrous medium such as vegetable oil (cottonseed oil, peanut oil, peanut oil, corn oil, corn oil, corn oil, corn oil). do not affect the therapeutic efficacy of the preparation and are not toxic in the amounts used (glycerol, propylene glycol, sorbitol). In addition, compositions can be advantageously prepared from which a solution is prepared immediately before use. Such compositions may contain liquid diluents, e.g., propylene glycol, diethyl carbonate, glycerol, sorbitol, etc., buffering agents, hyaluronidase, local anesthetics, and inorganic salts to provide the desired pharmacological properties. These compounds can also be combined with various pharmaceutically acceptable inert carriers, such as solid diluents, aqueous media, non-toxic organic solvents, and formed into capsules, tablets, lotions, troches, dry mixtures, suspensions, solutions, elixirs, and parenterals. . The compounds are generally used in various dosage forms in concentrations of from about 0.5% to about 90% by weight of the total composition.

68404 23

Comparative experiments R,? (“3> 2

0 V.X

5 Y '"' 'III According to the invention J \ OS0' 'R4 HO | 1 compound i! F · VS]' lx ^ 10 o OCH3 MIC, yug / ml 15 R = - * NH ~ R = NH? R = ^ - 'NH2 R = "' NH?

R = H R 1 = H R 1 = H 3 CC> R 1 = H

R ~ = H r3 = H R1 + R4 = R3 + R4 =

Microorganism R 1 = H R 2 = H -CO- -CO- 1) Staph, aur. 01A005 0.39 0.39 0.78 <0.10 2) "" 01A052 0.78 0.78 0.78 <0.10 3) "" 01A109 R> 200> 200> 200 200 20 4) "" 01A110 R> 200> 200> 200 200 5) "" 01A111 R - 0.39 <0.10 6) "" 01A087 RR> 200> 200> 200> 200 7) "" 01A400 R 3.12 6.25 25 6.25 8) Strp. fae. 02A006 0.39 1.56 0.78 £ 0.10 9) Strp. pyog. 02C203 <0.10 <0.10 0.20 <0.10 10) "" 02C020 R - 200 25 11) Myco. smeg. 05A001 100 200> 200 50 12) B. sub. 06A001 1.56 0.39 0.78 <0.10 13) E. coli 51A229 50 25 25 1.56 14) "" 51A266 50 50 25 1.56 15) "H 5IA125 R 50 50 25 3.12 16) Ps. Aerug. 52A104 200> 200> 200 100 17) Klebs. Pn. 53A009 200 100> 200 6.25 18) "" 53A031 R> 200 200> 200 6.25 iU 19) Erot. Mira. 57C064> 200> 200> 200 100 20) Prot. Norg. 57G001> 200> 200> 200 50 21) Salm. Chol-su. 58B242 50 200 50 3.12 22) Salm. Typhm. 58D009 50 100 100 3.12 23) "" 58D013-C 50 200 50 6.25 24) Past, multo. 59A001 1.56 1.56 0.39 <0.10 25) Serr. Mar 63A017 200> 200> 200 25 35 26) Ent. Aero 67A040 100> 200 100 6.25 27) Ent eloa 67B003 200> 200 100 12.5 28) Neiss. Sic. 66C000 0.39 6.25 <0.10 <0.10 68404 24 ^ (CH3> 2

s 0r \ x: O

RO,. „> 1 Reference compound

! Α ^ Λ [. L

10 (! ÖSO CH-.

6 'bcH3 1 J

15 Microorganisms R. = H

_ R1 = H

1) Staph, aur. 01A005 1.56 2) "" 01A052 1.56 3) "" 01A109 R> 50 4) "" 01AL10 R> 50 5) "" 01A111 R 0.39 2 0 6) "" 01A087 RR> 50 7) "" 01A400 R 50 8) Step. fae. 02A006 0.39 9) Strp. pyog. 02C203 0.20 10) "" 02C020 R> 50 11) Myco. smeg. 05A001> 50 12) B. sub. 06A001 0.20 13) E. coli 51A229> 50

Zb 14) "" 51A266> 50 15) "" 51A125 R> 50 16) Ps. Aerug. 52A104> 50 17) Klebs. pn. 53A009> 50 18) "" 53A031 R> 50 19) Prot. mira. 57C064 20) Prot. Morg. 57G001> 50 30 21) Verse. chol-Sun. 58B242 22) Ps. typhm. 58D009> 50

23) "" 58D013-C

24) Past, multo. 59A001 12.5 25) Serr. November. 63A017> 50 • 26) Ent. aero. 67A040> 50 27) Ent. life. 67B003> 50 35 28) Neiss. sic. 66C000 12.5 25 HO Λ qJ! “3’2 68404 ° '' V 5 R_o - .. L · .k ΰ of the invention. in accordance with 3>. γ "ο" 'Γ I prepared ίο A k L compound n "OCH-,,, 0 3 MIC, yug / ml 10 R = * NH- R = R = ^ NH R = ~ NH_ R, = HR, = HA Ri = H 2 R, + R = R_ + R = RT + R_ = Rl = CH-00 Micro-organisms_-CO -_- op-_z-co-J ις = hj 1) Staph, aur 01A005 £ 0.10 £ 0.10 0.39 3.12 2) "" 01A052 £ 0.10 £ 0.10 0.39 12.5 3) "" 01A110 R 200 200> 200> 200 15 4) "" 01A110 R 200 200> 200> 200 5) "" 01A111 R £ 0.10 £ 0.10 <0.1> 200 6) "" 01A087 RR 200 200> 200> 200 7) "" 01A400 R 3.12 3.12 3.12 25 8) Strp. fae. 02A006 £ 0.10 £ 0.10 0.39 50 9) Strp. pyog. 02C203 £ 0.10 £ 0.10 <0.10 1.56 10) "" 02C020 R - 11) Myoo. smeg. 05A001 50 50 100 0.39 20 12) B. sub. 06A001 £ 0.10 £ 0.10 £ 0.10 1.56 13) E. coli 51A229 3.12 1.56 3.12 100 14) "" 51A266 3.12 1.56 3.12 200 15) "" 51A125 R 3.12 3.12 6.25 200 16) Ps. Aerug. 52A104 50 100 200> 200 17 ) Klebs. Pn. 53A009 12.5 6.25 12.5 200 18) "" 53A031 R 6.25 6.25 50> 200 25 19) Prot. Mira. 57C064 200 100 200> 200 20) Prot. Morg. 57G001 50 50 200> 200 21) Salm. chol-su. 58B242 6.25 6.25 6.25> 200 22) Sal. typhm. 58D009 6.25 6.25 6.25> 200 23) "" 58D013-C 6.25 6.25 12.5 100 24) Past, mulch 59A001 £ 0.10 £ 0.10 £ 0.1 25 25) Serr Mar 63A017 25 25 50> 200 26) Ent Aero 67A040 6.25 6.25 50> 200 30 27) Ent eloa 67B003 25 25 25> 200 28) Neiss sic 66C000 £ 0.10 £ 0.10 £ 0.1 0.20

The results show that the compounds according to the invention are more active against a number of microorganisms than the reference compound.

35 The following examples are intended to illustrate the invention.

68404 26

Example 1 To a cooled mixture of 2'-acetyl-4 "-deoxy-4" -oxoerythromycin-A o -65 ° C with 3 ml of methylene chloride and 0.328 ml of dimethyl sulfoxide is added 0.652 ml of trifluoroacetic anhydride under nitrogen. After about a minute, a white slurry forms, indicating that a trifluoroacetic anhydride-dimethyl sulfoxide complex has formed. A solution of 1.0 g of 21-acetyleryl-romycin A * ethyl acetate obtained by crystallization from 2'-acetylerythromycin A in ethyl acetate in 7 ml of methylene chloride is added dropwise to the mixture, keeping the temperature at about -65 ° C. The mixture is stirred at about -60 ° C for 15 minutes, then cooled to -70 ° C. Triethylamine (1.61 mL) is added rapidly to the reaction mixture and the cooling bath is removed. The solution is stirred for 15 minutes, then added to 10 ml of water and the pH of the aqueous phase is adjusted to 10 ° C. The organic phase is separated, washed successively with water (3 x 10 ml) and brine (1 x 10 ml) and dried over sodium sulfate. The solvent is evaporated under reduced pressure to give 929 mg of crude product. Recrystallization from methylene chloride-hexane gives 329 mg of pure product, m.p. 105-108 ° C.

NMR (δ, CDCl 3): 3.28 (3H) s, 2.21 (6H) s and 2.3 (3H) s.

Similarly, using 2'-propionyl-erythromycin A-ethyl acetate as the starting material and following the above procedure, 2'-propionyl-4 "-deoxy-4" -oxoerythromycin-A is obtained.

Example 2

30 4 "-deoxy-4" -oxoerythromycin-A

A solution of 4.0 g of 2'-acetyl-4 "-deoxy-4" -oxoerythromycin-A in 75 ml of methanol is stirred at room temperature for 20 hours. The solvent is removed in vacuo and the residual white foam is crystallized from methylene chloride-hexane to give 3.44 g, m.p. 170.5-172.5 ° C.

68404 27 NMR (δ, CDCl 3) δ 3.36 (3H) s and 2.33 (6H) s.

An identical product is obtained when 2'-propionyl-4 "-deoxy-4" -oxoerythromycin-A is treated with methanol at room temperature.

5 Example 3

21-acetyl-4 "-deoxy-4" -oxo-erythromycin A

To a stirred solution of 13.7 g of 4 "-deoxy-4" -oxoerythromycin A in 100 ml of ethyl acetate is added 2.3 ml of acetic anhydride, and the resulting reaction mixture is stirred at room temperature for two hours.

The solution is added to 100 ml of water and the pH of the aqueous phase is adjusted to 9.5 with 6N sodium hydroxide solution. The organic layer is separated, dried over sodium sulfate and concentrated to give 14.5 g of a white foam which is recrystallized from methylene chloride-hexane. The compound obtained is identical to that obtained in Example 1.

Example 4 21-Acetyl-4n-deoxy-4 "-oxoerythromycin A-oxime To 20 500 ml of methanol is added 10.8 g of 2'-acetyl-4" -deoxy-4 "-oxoerythromycin-A, 1.94 g of hydroxylamine hydrochloride and 11.0 g of barium carbonate, and the resulting suspension are stirred at room temperature for 3.5 hours, filtered and the filtrate is evaporated under reduced pressure, the resulting foam is dissolved in ethyl acetate and the solution is extracted with pH-adjusted water. The organic phase is separated, dried over sodium sulfate and evaporated in vacuo to give 10.6 g of the desired product NMR (ε, CDCl3): δ 3.33 (3H) s, 2.30 (6H) s and 2 .06 (3H) s.

Example 5 2'-Acetyl-4 "-deoxy-4" -oxoerythromycin A-O-acetyloxime

To a solution of 330 mg of 2'-acetyl-4 "-deoxy-35 4" -deoxy-4 "-oxoerythromycin A-oxime in 30 ml of 68404 28 ethyl acetate is added, with stirring, 64.2 μl of acetic anhydride, and 15.8 .mu.l of acetic anhydride and 23.4 .mu.l of triethylamine are added and stirring is continued for 4 to 5 hours, the reaction mixture is then added to water and the pH of the mixture is adjusted to about 9.0, the ethyl acetate layer is separated, dried over sodium sulfate and dried. evaporate to dryness in vacuo to give 300 mg of the desired product NMR (Sf CDCl3): 3.38 (3H) s, 2.25 (6H) s, 2.20 (3H) s, 2.05 (3H) s and 1.56 (3H) s.

Correspondingly, using 2'-propionyl-4 "-deoxy-4" -oxoerythromycin A and 4 "-deoxy-4" -oxoerythromycin A-oxime in the above method, 2'-acetyl-4 "-deoxy-4" - the corresponding O-acetyl derivatives are obtained from oxoerythromycin A-oxime.

Example 6 2'-Acetyl-4 "-deoxy-4" -aminoerythromycin-A A mixture of 14.0 g of 21-acetyl-4 "-deoxy-4 '' - oxoerythromycin A-O-acetyloxime and 60 g of large -20 propane-washed Raney nickel in 400 ml of isopropanol, stirred under an atmosphere of hydrogen at an initial pressure of 6.9 MPa overnight at room temperature, the catalyst is filtered off and the filtrate is evaporated, the residual white foam is dissolved in 400 ml of isopropanol and a further 50 g of batch of isopropanol freshly washed

Raney-nickel. The hydrogenation is continued overnight at room temperature with an initial hydrogen pressure of 6.9 MPa. The catalyst is filtered off and the filtrate is evaporated to dryness in vacuo to give the desired product.

Example 7 Starting from the appropriate O-acetyloxime and following the procedure of Example 6, the following 4 "-aminoerythromycin A analogs are prepared: 68404 29 R, \ N (CH): 0 3 2 Ογ ^ 5 mn„ y

I J! 0 I

ho — L A.

A f ° ·: / ° Y 0 ^ X ^ H2

X OCIL

10 3 fl 15 H-CH3CH2C-Example 8

4 4 "-deoxy-4" -aminoerythromycin-A

A solution of 2.17 g of 2'-acetyl-4 "-deoxy-4" -aminoerythromycin-A in 50 ml of methanol is stirred at room temperature overnight. The solvent is removed under reduced pressure and the residual foam is treated with a mixture of chloroform (50 ml) and water (50 ml). The pH of the aqueous layer is adjusted to 9.5, and the organic layer is separated. The chloroform layer is treated with a new portion of water and the pH is adjusted to 4.0. The pH of the aqueous layer containing the acidic product is adjusted by gradually adding base 30 to 5, 6, 7, 8 and 9, and already at this pH the mixture is extracted with a new batch of chloroform. The extracts obtained at pH 6 and 7 contain most of the product, and these extracts are combined, water is added and the pH is adjusted to 4. The pH of the aqueous layer is again adjusted to 5, 6 and 7, and at each pH the extract is extracted with chloroform. The chloroform extract obtained at pH 6 is dried over sodium sulfate and concentrated to give 249 g of product as an epimer mixture.

NMR (ε, CDCl 3): 3.30 (1H) s, 3.26 (2H) s, 2.30 (6H) s and 1.46 (3H) s.

In a similar manner, 4 "-deoxy-4" -aminoerythromycin-A is prepared by solvolyzing 2'-propionyl-4 "-deoxy-4" -aminoerythromycin-A.

Example 9 4 "-deoxy-4" -aminoerythromycin A To a stirred solution of 3.0 g of 4 "-deoxy-4" -oxoerythromycin A in 30 ml of methanol is added 3.16 g of dry ammonium acetate under nitrogen. After 5 minutes, 188 mg of sodium cyanoborohydride are rinsed into the reaction mixture with 5 ml of methanol, and the reaction mixture is stirred overnight at room temperature. The pale yellow solution is poured into 300 ml of water and the pH is adjusted to 6 ° C. The aqueous phase is extracted at pH 6, 7, 7.5, 8, 9 and 10 with 125 ml of diethyl ether for extraction. The extracts obtained at pH 8, 9 and 10 are combined and washed with 125 ml of water. The separated aqueous layer is extracted with ether (1 x 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 ethyl acetate (1 x 100 mL) at pH 8, 9 and 10.

The ethyl acetate extracts obtained at pH 9 and 10 are combined, washed with brine and dried over sodium sulfate. Evaporation of the solvent in vacuo gives 30 mg of an epimeric mixture of the desired product as an ivory foam.

Example 10 4 "-deoxy-4" -aminoerythromycin A (second epimer) A solution of 10.0 g of an epimer mixture of 21-acetyl-4 "-deoxy-4" -aminoerythromycin-A in 150 ml of methanol is stirred at room temperature. at 35 ° C for 72 hours. The solvent is removed in vacuo and the residue Liu is taken up in a stirred mixture of water (150 ml) and chloroform (200 ml) 68404 31. The aqueous layer is discarded and a new 150 ml portion of water is added. The pH of the aqueous layer is adjusted to 5 ° C, and the chloroform layer is separated. The pH of the aqueous phase is then adjusted to 5.5, 6, 7, 8 and 9 and extracted after adjustment with 100 ml of chloroform. The extracts obtained at pH 6, 7 and 8 are combined, washed successively with water and saturated brine and dried over sodium sulfate. Evaporation of the solvent under reduced pressure gives 2.9 g of an epi-10 meri mixture of 4 "-deoxy-4" -aminoerythromycin A. A 1.9 g sample of the mixture is triturated in diethyl ether to crystallize some of the insoluble foam. The solid is filtered and dried to give 67 mg of the second epimer of 4 "-deoxy-4" -aminoerythromycin A, m.p. 140-147 ° C.

Example 11 11,2'-Diacetyl-4 "-deoxy-4" -oxoerythromycin A-6,9-hemiketal

A solution of 10 g of 2'-acetyl-4 "-deoxy-4" -oxoerythromycin-A in 250 ml of pyridine is treated with 40 ml of acetic anhydride and the reaction mixture is allowed to stand at room temperature for 10 days. Most of the solvent is removed in vacuo, and the remaining concentrate is taken up in a mixture of water (150 ml) and chloroform (100 ml). The pH of the aqueous phase is raised to 9.0 and the chloroform is separated off, dried over sodium sulfate and evaporated to dryness.

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

Example 12 Using the appropriate 4 "-deoxy-4" -oxoerythromycin A and the required alkanoic anhydride as starting materials and following the procedure of Example 11, the following compounds are prepared: 68404 32, R1 N (CH_) _ “χλ \ λ! 2

Η ° -; L

N CX

Example 11 11-acetyl-4 "-deoxy-4" -oxoerythromycin A-6.9-hemiketal. 'cch3 10 [mu] o ch3c- ch3ch2c- o o 15 "11 CH3CH2C-CH3CH2C-

A solution of 3.0 g of 11,2'-diacetyl-4 "-deoxy-4" -oxoerythromycin A-9,6-hemiketal in 50 ml of methanol is stirred overnight under nitrogen. The solvent is removed in vacuo to give the desired is obtained as a yellow foam (3.0 g).

NMR (δ, CDCl 3): 3.35 (3H) δ, 2.31 (6H) δ, 2.13 (3H) and 1.55 (3H) s.

In a similar manner, by the method of Example 13, the compounds of Example 12 were converted to 11-acetyl-4 "-deoxy-4" -oxoerythromycin-A-6,9-hemiketal and 11-propionyl-4 "-deoxy-4" -oxoerythromycin-A-6 , 9-hemiketal.

Example 14 35 11-Acetyl-4 "-deoxy-4" -aminoerythromycin A-6.9-hemiketal 68404 33

To a stirred solution of 4.4 g of acetyl-4 "-deoxy-4" -oxoerythromycin A-6,9-hemiketal and 4.38 g of ammonium acetate in 75 ml of methanol is added 305 mg of 85% sodium cyanoborohydride. After stirring at room temperature overnight, the reaction mixture is poured into 300 ml of water and 250 ml of chloroform are added. The pH of the aqueous layer is adjusted to 9.8, and the chloroform layer is separated. The aqueous layer is re-extracted with chloroform, and the chloroform extracts are combined, dried over sodium sulfate and evaporated to give a white foam. The residual foam is dissolved with stirring in a mixture of water (125 ml) and chloroform (125 ml) and the pH is adjusted to 5, 6, 7 and 8 and extracted with chloroform after each adjustment. The extracts obtained at pH 6 and 7 are combined, washed with saturated brine and dried over sodium sulfate. Evaporation of the solvent gives 1.72 g of the desired product as a white foam. The product is dissolved in as little diethyl ether as possible, and then hexane is added to the solution until it begins to cloud. The crystalline product formed is filtered off and dried, yielding 1.33 g of product, m.p. 204.5-206.5 ° C.

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

25 Example 15

By repeating the procedure of Example 14 using the appropriate 4 "-deoxy-4" -oxoerythromycin A and isopropanol instead of methanol as the solvent, the following compounds are obtained: 68404 34, R1 MCH,), ί0 ^ Λ ^ Λ «2 ° * ,,,> 5 HO '“^ L o pL x ^' fy (j yf: m2 /" 'cc * 3 10 Rj ^ Rj oo

It II

CH-jC-CH3C "

O O

CH, C-CH-.CH „C-i0 0 0 ch3ch2c- ch3ch2c- 0 o CH3CH2C ch3c ~ ^ Example 16 2'-Acetylerythromycin A-6,9-hemiketal-11,12-carbonate ester

To a solution of 13.2 g of erythromycin A-6,9-hemiketal-11,12-carbonate ester (U.S. Patent 3,417,077) in 25,150 ml of benzene is added 1.8 ml of acetic anhydride, and the reaction mixture is stirred. at room temperature for 1.5 hours. The solution is poured into 200 ml of water and the aqueous phase is basified to pH 9. The benzene layer is separated, dried over sodium sulfate and evaporated to dryness to give 15.3 g of a white foam. This foam is triturated in 50 ml of diethyl ether to crystallize. Filtration and drying gives 12.6 g of pure product, m.p. 224.5-228.5 ° C.

NMR (δ, CDCl 3): 3.36 (3H) s, 2.30 (6H) s, 2.06 (3H) s and 1.61 (3H) s.

35 68404

Similarly, following the procedure of Example 16, substituting an equivalent amount of propionic anhydride for acetic anhydride, 21-propionylerythromycin A-6,9-hemiketal-11,12-5 carbonate ester is obtained.

Example 17 21-Acetyl-4 "-deoxy-4" -oxoerythromycin A-6,9-hemiketal-11,12-carbonate ester in a suspension cooled to -5 ° C with 6.19 g of N-chlorosuccinimide in 150 ml in toluene and 50 ml of benzene, 4.46 ml of dimethyl sulfide are added. After stirring for 20 minutes, the suspension is cooled to -25 [deg.] C. and 12.4 g of 2'-acetylerythromycin A-hemiketal-11,12-carbonate ester 15 partially dissolved in 80 ml of toluene are added dropwise. During the addition, the temperature is maintained at -19 to -25 ° C and then at -25 ° C for a further two hours. 6.79 ml of triethylamine are then added in one portion. The cooling bath is removed and the temperature is allowed to rise to -10 ° C. The reaction mixture is then poured into water, and the pH of the aqueous phase is adjusted to 8.4 to 9.0. The organic layer was separated, dried over sodium sulfate and evaporated in vacuo to give a white foam (14.0 g). Trituration of this foam in diethyl ether causes it to crystallize. The product 25 is filtered and dried to give 11.3 g of crystalline material, m.p. 212-213.5 ° C.

NMR (δ, CDCl 3): 5.26 (1H) t, 3.36 (3H) s, 2.30 (6H) s, 2.13 (3H) s, 1.63 (3H) s and 1.50 (3H) s.

Similarly, following the procedure of Example 17 and using an equivalent amount of 2'-propionylerythromycin A-6,9-hemiketal-11,12-carbonate ester instead of 2'-acetyl ester, 2'-propionyl-4 "-deoxy-4 * - oxoerythromycin A-6,9-hemiketal-11,12-carbonate ester.

68404 36

Example 18 4 "-deoxy-4" -oxoerythromycin A-6,9-hemiketal-11,12-carbonate ester 42.9 g of 2'-acetyl-4 "-deoxy-4" -oxoerythromycin-5-n-A-6 The 9-hemiketal-11,12-carbonate ester is added to 800 ml of methanol, and the solution is stirred at room temperature for 72 hours. The solvent is removed in vacuo to give 41 g of product as a white foam. It is dissolved in about 100 ml of acetone, and water is carefully added to the solution until precipitation begins. The resulting crystalline precipitate is stirred for 40 minutes, then filtered and dried to give 34.2 g of the desired product, m.p. 186.5-183 ° C.

NMR (ε, CDCl 3): 5.66 (1H), 3.35 (3H) s, 2.35 (6H) s, 1.65 (3H) s and 1.51 (3H) s.

In a similar manner, the same product is obtained when an equivalent amount of 2'-propionyl-4 "-deoxy-4" -oxoerythromycin A-6,9-hemiketal-11,12-carbonate ester is used in the above process instead of 2'-acetyl ester.

Example 19 4 "-deoxy-4" -aminoerythromycin A-6,9-hemiketal-11,12-carbonate ester to 189 g of 4 "-deoxy-4" -oxoerythromycin A-6,9-25 hemiketal-11, 12-Carbonate ester in 1200 ml of methanol is added with stirring at room temperature to 193 g of ammonium acetate. After 5 minutes, the resulting solution is cooled to about -5 ° C and then treated for 45 minutes with 13.4 g of 85% sodium cyano-30 borohydride in 200 ml of methanol. The cooling bath is removed and the reaction mixture is stirred at room temperature overnight. The reaction mixture is evaporated in vacuo to 800 ml and added with stirring to a mixture of water (1800 ml) and chloroform (900 ml). The pH is adjusted to 6.2-4.3 with 6N hydrochloric acid, and the chloroform layer is separated. The chloroform layer is poured into 1 liter of water and the pH is adjusted to 37 68404 95. The organic phase is separated, dried over sodium sulphate and evaporated under reduced pressure to give 174 g of a white foam. This is dissolved in a mixture of water (1 L) and ethyl acetate (500 mL) and the pH is adjusted to 5.5. The ethyl acetate layer is separated, and the pH of the aqueous layer is adjusted successively to 5.7 and 9.5, and after each pH adjustment, 500 ml of ethyl acetate are extracted. The ethyl acetate extract obtained in 9.5 is dried over sodium sulphate and evaporated to dryness in vacuo to give 130 g of a foam. 120 g of this foam are dissolved in a mixture of water (1 L) and methylene chloride (1 L). The pH of the aqueous layer is adjusted successively to 4.4, 4.9 and 9.4 and extracted after adjustment with 1 liter of methylene chloride. The methylene chloride extract obtained at pH 9.4 is dried over sodium sulphate and evaporated to dryness under reduced pressure to give 32 g of product in the form of a white foam. Crystallization from 250 ml of an acetone-water mixture (1: 1, v / v) gives 28.5 g of crystalline epimers.

NMR 100 MHz (ε, CDCl 3): 5.20 (1H) m, 3.37 (1.5) s, 3.34 (1.5H) s, 2.36 (6H) s, 1.55 ( 3H) s and 1.41 (3H) s.

Example 20 Separation of 4 "-deoxy-4" -aminoerythromycin A-6,9-hemiketal-11,12-carbonate ester epimers

A high pressure liquid chromatography column (1.3 x 259 cm) packed with formamide-impregnated GF 254 silica gel and eluted with chloroform is charged with 200 mg of the epimer mixture. At a pressure of 1.65 MPa, 10 ml and boils are collected at a rate of 4.76 ml / min. Fractions 14-21 and 24-36 are recovered.

Fractions 14 to 21 are combined and concentrated to about 50 ml. 50 ml of water are added and the pH is adjusted to 9.0. The chloroform layer is separated, dried over sodium sulfate and evaporated to give 106 mg of a white foam. Trituration with diethyl ether gives a foam crystallization. The crystalline product is stirred at room temperature for 1 hour, then filtered and dried to give 31.7 mg of 31.7 mg, m.p. 194-196 ° C.

NMR 100 MHz (ε, CDCl 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.

Fractions 24-36 are combined and combined and treated in the same manner as above to give 47.1 mg of product as a white foam identical to that obtained in Example 25.

Example 21

To a suspension of 11.1 g of 2'-acetyl-4 "-10-deoxy-4" -oxoerythromycin A-6,9-hemiketal-11,12-carbonate ester in 300 ml of isopropanol is added 10.7 g at room temperature with stirring. g of ammonium acetate. After 5 minutes, 747 mg of sodium cyanoborohydride in 130 ml of isopropanol are added to the reaction mixture over 30 minutes, and the reaction mixture is stirred at room temperature overnight. The pale yellow solution is poured into 1,100 ml of water, to which is added 400 ml of diethyl ether. The pH is adjusted to 4.5 and the ether layer is separated. The aqueous layer is adjusted to pH 9.5 and extracted (2 x 500 mL) with chloroform. The chloroform extracts are combined, dried over sodium sulfate and concentrated to give 7.5 g of a yellow foam. Recrystallization from diethyl ether gives 1.69 g, which is recovered as well as the mother liquors for further work-up.

To the mother liquors obtained above, 75 ml of water are added and the pH is adjusted to 5.0. The ether layer is replaced with a new 75 ml portion of ether and the pH is adjusted to 5.4. The ether is replaced with ethyl acetate and the pH is raised to 30. The basic aqueous layer is extracted (2 x 75 ml) with ethyl acetate, and the first ethyl acetate extract is dried over sodium sulfate and evaporated to dryness. The residual foam (1.95 g) is added to a mixture of water (75 ml) and diethyl ether (50 ml) and the pH is adjusted to 5.05. The ether is separated off and the pH of the aqueous layer is adjusted to 5.4, 6.0, 7.05 and 8.0 in succession, and after each pH adjustment it is extracted with 50 ml of diethyl ether. Finally, the pH is adjusted to 9.7 and the aqueous layer is extracted with 50 ml of ethyl acetate. To the ether extract obtained at pH 6.0, 75 ml of water are added and the pH is adjusted to 9.7. The ether layer is separated, dried and evaporated in vacuo to give 460 mg of a white foam.

NMR 100 MHz (δ, CDCl 3): 5.20 (1H) t, 3.43 (2H) s, 3.40 (1H) s, 2.38 (6H) s, 2.16 (3H) s, 1.70 (3H) s and 1.54 (3H).

NMR values indicate that the product obtained consists of 2'-acetyl-4 "-deoxy-4" -aminoerythromycin A-6,9-hemi-ketal-11,12-carbonate ester epimers.

1.69 g of the product obtained above are dissolved in a mixture of water (75 ml) and diethyl ether (75 ml) and the pH is adjusted to 4.7. The ether is separated and the aqueous layer is extracted with a new portion of 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 are dried over sodium sulfate and evaporated under reduced pressure to give 1.26 g of a white foam as a residue. Crystallization gave 411 mg of product, m.p. 193-196 ° C (decomposes). The mother liquor is evaporated to dryness and the residue is dissolved in hot ethyl acetate. The residue is allowed to stand at room temperature overnight. The precipitated crystalline material is filtered off and dried, yielding 182 mg of additional product, m.p. 198-202 ° C (decomposes).

NMR 100 MHz (δ, CDCl 3): 5.10 (1H) t, 3.34 (2H) s, 3.30 (1H) s, 2.30 (6H) s, 2.08 (3H) s, 1.62 (3H) s and 1.48 (3H) s. NMR values indicate that the product obtained consists of 2'-acetyl-4 "-deoxy-4" -aminoerythromycin A-11,12-carbonate ester epimers.

In a similar manner, repeating the procedure of Example 21 starting from 2, -propionyl-4 "-deoxy-4" -35 oxoerythromycin A-6,9-hemiketal-11,12-carbonate ester, 2'-propionyl-4 "-deoxy is obtained. -4 "-aminoeryt-68404 40 romycin A-6,9-hemiketal-11,12-carbonate ester and 2'-propionyl-4" -deoxy-4 "-aminoerythromycin A-11,12-carbonate ester.

Example 22 A solution of 400 mg of 2'-acetyl-4 "-deoxy-4" -aminoerythromycin A-6,9-hemiketal-11,12-carbonate ester in 20 ml of methanol is stirred at room temperature overnight. . The reaction solution is poured into 100 ml of water, then 50 ml of ethyl acetate are added.

The pH is adjusted to 9.5 and the organic phase is separated.

The extraction is repeated with a further 50 ml of ethyl acetate. The combined ethyl acetate extracts are dried over sodium sulfate and evaporated to give 392 mg of a white foam. By trituration in diethyl ether and crushing with a glass rod, it is made to crystallize. After standing for 30 minutes at room temperature, the crystals are filtered off and dried to give 123 mg of product which is identical in NMR values to the product obtained in Example 24.

NMR 100 MHz (δ, CDCl 3): 3.26 (3H) s, 2.32 (6H) s, 1.61 (3H) s and 1.44 (3H) s.

NMR values indicate that this crystalline product is the second epimer of 4 "-deoxy-4" -aminoerythromycin A-11,12-carbonate ester.

The mother liquor obtained from the above crystallization is evaporated in vacuo to give 244 mg of a white foam. It is identical to the product obtained in Example 19.

NMR values indicate that this product consists of 4 "-deoxy-4" -aminoerythromycin A-6,9-hemiketal-11,12-carbonate ester epimers and is identical to the product of Example 19.

Example 23

By a method similar to Example 35 22, 2'-propionyl-4 "-deoxy-4" -aminoerythromycin A-6,9-hemiketal-11,12-carbonate-68404 41 thiester 4 "-deoxy-4" -aminoerythromycin is obtained by methanolysis. -A-11,12-carbonate ester and 4 "-deoxy-4" -aminoerythromycin-A-6,9-heme ± ketal-11,12-carbonate ester.

Example 24 5 g of the non-crystalline 4 "-deoxy-4" -aminoerythromycin A-11,12-carbonate ester-epimer mixture obtained in Example 19 are dissolved in 50 ml of diethyl ether. The product is crystallized by scraping with a glass rod. The crystalline product is stirred for 20 minutes, then filtered and dried to give 1.91 g, m.p. 198.5-200 ° C.

NMR 100 MHz (g, CDCl 3): 3.26 (3H) s, 2.30 (6H) s, 1.61 (3H) s and 1.45 (3H) s.

NMR values indicate that the crystalline product is the second epimer of 4 "-deoxy-4" -aminoerythromycin A-11,12-carbonate ester, which is identical to the ketone compound of Example 22.

Example 25 1 g of the epimer obtained in Example 24 is dissolved in 20 ml of acetone and the solution is heated on a steam bath until it begins to boil. 25 ml of water are then added and the solution is stirred at room temperature. After stirring for 1 hour, the precipitate formed is filtered off and dried, yielding 581 mg of product, m.p. 147-149 ° C.

NMR 100 MHz (ε, CDCl 3) δ 5.12 (1H) d, 3.30 (3H) s, 2.30 (6H) s, 1.62 (3H) s and 1.36 (3H) s.

NMR values indicate that this is the second epimer of 4 "-deoxy-4" -aminoerythromycin A-6,9-hemiketal-11,12-carbonate thiester, which is identical to the epimer obtained from fractions 24-36 of Example 20.

30 Example 26

4 "-deoxy-4" -amino-A

20 g of 4 "-deoxy-4" -oxoerythromycin A, 31.6 g of ammonium acetate and 10 g of 10% palladium on carbon in 200 ml of methanol are shaken under an atmosphere of hydrogen at an initial pressure of 0.34 MPa overnight. The catalyst is filtered off and the filtrate is evaporated to dryness in vacuo. The residue is partitioned between 68404 42 water and chloroform at pH 5.5. The aqueous layer is separated, the pH is adjusted to 9.6, then chloroform is added. The organic layer is separated, dried over sodium sulfate and evaporated to dryness under reduced pressure.

The residual white foam (19 g) is triturated in 150 ml of diethyl ether at room temperature for 30 minutes. The resulting solid is filtered and dried to give 9.45 g of a second epimer identical to that obtained in Example 10.

The diethyl ether filtrate is evaporated to dryness to give 6.89 g of product containing the second epimer and some impurities.

Example 27 4 "-deoxy-4" -aminoerythromycin A A 2 g of 4 "-deoxy-4, '- oxoerythromycin A, 3.1 g of ammonium acetate and 2.0 g of Raney nickel in 50 ml of methanol shake under an atmosphere of hydrogen at an initial pressure of 0.34 MPa at room temperature overnight, add a further 3.16 g of ammonium acetate and 2.0 g of Raney nickel and continue the hydrogenation for a further five hours, filter the solids and evaporate the filtrate to dryness in vacuo. The aqueous phase is separated, the pH is adjusted to 9.6, then chloroform is added, the chloroform-25 extract is separated, dried over sodium sulfate and evaporated under reduced pressure to give 1.02 g of product as a yellow solid. as its foam. The predominant isomer at the 4 "position has the opposite configuration to the product of Example 10.

Example 28 2'-Acetyl-4 ', - deoxy-4 "-aminoerythromycin-B To a solution of 4.5 g of 2'-acetyl-4" -deoxy-4 "-oxoerythromycin-B (U.S. Patent 3,884,903) in 45 ml of isopropanol, 4.66 g of dry ammonium acetate are added with stirring under a nitrogen atmosphere, after 10 minutes 376 mg of sodium cyanoborohydride are added by rinsing in 10 ml of isopropanol, and the reaction mixture is stirred at room temperature overnight. The resulting pale yellow solution is poured into 400 parts of water and the pH is adjusted to 6.0 The aqueous layer is extracted at pH 6, 7, 7.5, 5, 8, 9 and 10 using 250 ml of diethyl ether for each extraction. and 10 The combined extracts are washed and washed with 250 ml of water The separated aqueous layer is extracted with ether (1 x 100 ml) at pH 7, ethyl acetate (1 x 100 ml) at pH 7, ether (1 x 100 ml) at pH 7.5 with ether (1 x 100 ml). x 100 ml) at pH 7.5, ethyl acetate (1 x 100 ml) at pH 7.5 and ethyl acetate (1 x 100 ml) at pH 8, 9 and 10. At pH 9 and The resulting ethyl acetate extracts were combined, washed, brine and dried over sodium sulfate. Solution 15 is removed in vacuo to give an epimer mixture of the desired product as a cream foam.

In a similar manner, 4 "-deoxy-4" -aminoerythromycin-B is prepared from 4 "-deoxy-4" -oxoerythromycin-B.

Example 29 4 "-deoxy-4" -aminoerythromycin-B A solution of 4.34 g of 2'-acetyl-4 "-deoxy-4" -aminoerythromycin-B in 100 ml of methanol is stirred overnight at room temperature. temperature. The solvent is removed under reduced pressure and the residual foam is treated with a mixture of chloroform (100 ml) and water (100 ml). The pH of the aqueous layer is adjusted to 9.5, and the organic layer is separated. A new portion of water is added to the chloroform layer and the pH is adjusted to 4.0. The pH of the acidic aqueous layer containing the product is gradually adjusted by adding base to 5, 6, 7, 8 and 9 and extracted at each pH with chloroform. The extracts obtained at pH 6 and 7 contain most of the product and are combined and treated with water at pH 4. The pH of the aqueous layer is again adjusted to 5, 6 and 7 and extracted at each pH with chloroform. The chloroform extract 68404 44 obtained at pH 6 is dried over sodium sulfate and evaporated to dryness to give the desired product as an epimer mixture.

Example 30 4 "-deoxy-4" -aminoerythromycin A-6,9-hemiketal-5,11,12-carbonate ester aspartate in a solution heated to 40 ° C with 1.0 g of 4 "-deoxy-4" - aminoerythromycin A-6,9-hemiketal-11,12-carbonate ester in 6 ml of acetone, 20 ml of water are added followed by 175 mg of L-Aspartic acid. The mixture 10 is heated to reflux for 1.5 hours, then filtered hot. The filtrate is evaporated from acetone and the concentrated filtrate is dry cooled to give 1.1 g of the desired product as a white solid.

Example 31 4, '- Deoxy-4 "-aminoerythromycin A-6,9-hemiketal II, 12-carbonate ester dihydrochloride 7.58 g of 4" -deoxy-4 "-aminoerythromycin A-6,9- hemiketal-11,12-carbonate ester in 50 ml of dry ethyl acetate, 20 ml of 1N HCl-ethyl-acetate solution are added, and the resulting solution is evaporated to dryness under reduced pressure, and the residue is triturated with ether, filtered to give the desired salt.

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

Example 32 4 "-deoxy-4" -aminoerythromycin A-6,9-hemiketal-11,12-carbonate ester hydrochloride

The procedure of Example 31 is repeated except that 10 mL of 1 N HCl-ethyl acetate solution is added. The solution 30 is evaporated to dryness, the residual mono-hydrochloride disalt salt is triturated in ether and filtered.

Similarly, other amine compounds of the invention are converted to their monoacid addition salts.

Claims (6)

68404 1. N (CH) _ p,, - Vs · vSVi1 10 R ^ 0, Y ° X ° ui R3 ° [V / V R3 ° ~ Ol ° o / <^ ™ 2 A 'O * 1 * / 0CH3 2 Λχ, Λ 15 111 IV 3 wherein and R ^ represent a hydrogen atom or An alkanoyl group having 2 to 3 carbon atoms is an alkanoyl group having 2 to 3 carbon atoms, R 1 is a hydrogen atom or R 20 3a R 3 together form a group -C- or R 3 and R 2 together form a group -C-, as well as for the preparation of their pharmaceutically acceptable acid addition salts, characterized in that the compound of the formula R 1 \ N (CH 2 O 0 = O 3 3 .1 N (CH 7), ΓΧ H0A ^ kil H0 11 o / r 2 ° „'A. oAqA 30. or ^ "f 1 H ° (VT A / R3 ° -k. A * X '0CH, o / *' ·, Y" A process for the preparation of therapeutically useful 4 "-deoxy-4" -aminoerythromycin A derivatives of the formula Process according to Claim 1, characterized in that Y "is NH and Y 1 is NH, N - OH or N - OCOCH 4 and in that the reduction is a catalyst reduction. Process according to Claim 3, characterized in that the reduction is carried out with hydrogen in the presence of Raney nickel, palladium on carbon or platinum oxide. 3 O / OCH 35 68404 wherein Y 'and Y "are NH, N - OH or N-OCOCH2, the reduction is carried out, wherein a) when Y" is NH and Y' is NH, N - OH or N-OCOCl2, the reduction is carried out catalytically by hydrogenation and 5 b) when Y 'or Y "is NH prepared in situ from the corresponding ketone of formula Λ \ N (CU,),: OI 3 2, flCHJ,» vs -yS “vA \ A 10 HO NX n ^ A vhol "'I 1 Γ H0 1 or ^ T 1 V °" · / Υ 3 vi lC / γ vi 1V v 15 3' OCH I II 3 by condensing said ketone with an ammonium salt of an alkanoic acid, the reduction is carried out by reduction with a hydride, and if desired c) when R 1 or a hydrogen atom is converted into an alkanoyl group and / or when R 1 or R 2 is an alkanoyl group it is converted into a hydrogen atom, and if desired d) pharmaceutically acceptable salts are formed. Process according to Claim 1, characterized in that Y * and Y "both represent an imino group and an excess of ammonium salt of alkanoic acid 35, preferably ammonium acetate, is used. 68404 Process according to Claim 4, characterized in that the reducing agent is sodium cyanoborohydride. p. 68404