FI68059B - FOERFARANDE FOER FRAMSTAELLNING AV TERAPEUTISKT ANVAENDBARA HAVSYNTETISKA 4 '' - AMINO-OLEANDOMYCINDERIVAT - Google Patents

Description

1 68059

Process for the preparation of therapeutically useful semisynthetic M "-amino-oleandomycin derivatives

Separated from application 780355 (patent 65262) 5

Oleandomycin, its production using broth, and its use as an antibacterial agent are first described in U.S. Patent No. 2,757,123. The naturally occurring compound is known to have the following structure: 8a Hv fCH3} 2 HOV "10 15 I'll 5I" 6 '> 13 3 <6 ”" k - - ° .........

1 fl l h "o 2" s4 ^ j, //// oh 20 '0CH3

The conventionally accepted numbering scheme and stereochemical representation of oleandomycin and related compounds are indicated in their various positions.

Several synthetic variants of this compound are known, in particular those in which one, two or three of the free hydroxyl groups observed at positions 2 1, 4 "and 11 are esterified to acetyl esters. In addition, U.S. Pat. No. 3,022,219 describes similar variants. those in which acetyl in the abovementioned esters has been replaced by another, preferably unbranched, lower alkanoyl having from three to six carbon atoms.

The invention relates to a process for the preparation of therapeutically useful 4 "-amino-Olean-35 domycin derivatives of the formula IV, V or VI and their pharmaceutically acceptable acid addition salts, 2,68059; νΛ Ά'2 s, n (ch,

- f - ‘f S | 'o I

s Rv r Y \ X

i A R0 Y 'A

v'j C ti n r ° ...... <^ x S | te3 Y '' Ά OCH3 i | ! * 2 10 M 0GH9 IV V 3 N (CH „) 0 - Ri n I 3 2 ° Ύ <Vk RO„ „/ 15>; 11 "Ύ; nh2 0 OCHo

20 VI

wherein R and represent hydrogen or alkanoyl having two or three carbon atoms; and R 1 is hydrogen or

O

Alkyl of 1 to 6 carbon atoms.

The compounds of formula IV form the most preferred group in this class of chemotherapeutic agents. Particularly preferred in this group are compounds wherein R 2 is hydrogen and R is acetyl. Compounds of formulas V and VI wherein R is acetyl are also preferred.

30 Compounds V and VI, although all derived from naturally occurring oleadomycin, differ in structure from position 8. In natural matter and in compound IV, this structure is an epoxide ring similar to the following: 3 68059

Compounds related to Compound V contain a methyl group at position 8 with its associated stereochemical structures and are described as follows:

5 ° yS

The oleadomycin variants of formula V are termed 8,8a-10 deoxy-8,8a-dihydro-oleandomycins.

The compounds of formula VI which contain a cyclopropyl ring in position 8 are designated 8,8a-deoxy-8,8a-methyleneaminomomycins and are described as follows: 15 ° Yh 20 The compounds of formulas IV, V and VI are prepared in such a way that i ) reducing a compound of formula I, II or III, R, N (CHJ0 1 \ 6 1 N (CH-) 0 0 XT '' 0 1 r 3 l 2 5 Y v 1 '' ό Λ R0 "r ' ν-σ ^ ο-ν f)> i

1 L

'1 X X

CL / X! J X] r "" 0 „.rO · ^

30 O L X C> V | X '' X

° ocHj ii rX

1 * ls * lat3> 2 11 0CH3

Y is Vs-% y ^ oy li tl T ^ x ITI 0CH3, 68059 wherein R and are each hydrogen or alkanoyl having two or three carbon atoms and X is NR, N-OH, O-OCH2 or N-OC- CHg such that 0 a) a) when X is N-OH, N-OCH4 or N-OC-CH2, said reduction is carried out by catalytic hydrogenation and b) when X is NR4, this imino group is formed from the corresponding compounds of formulas I, Of ketones II or III (X = O) by condensing said ketone with an amine or ammonium salt of a lower alkanoic acid or an amine or ammonium salt of an inorganic acid, and the reduction which is a hydride reduction is carried out in situ, and if desired ii) when R or R 1 is hydrogen, they are converted to alkanoyl, respectively, and / or when R 1 or R 2 is alkanoyl, they are converted to hydrogen, respectively; and / or iii) forming pharmaceutically acceptable

The acid addition salts. O

The reduction of ketone derivatives (X = N-OH, N-OCH 4 or N-OCCH 2) is carried out by catalytic hydrogenation in which an oxime or derivative dissolved in a lower alkanol such as isopropanol and Raney nickel, 10% palladium The carbon mixture or platinum oxide catalyst is shaken in hydrogen gas at room temperature overnight at an initial pressure of 50 pounds per square mile (about 3.5 kg / cm). The spent catalyst is filtered off, after which the desired antibacterial 4 "-deoxy-4" -amino-substituted product of formula IV, V or VI is isolated by removing the solvent from the filtrate. If methanol is used as the solvent in the reduction, solvolysis of the 2'-alkanyl group takes place. To avoid detachment of this part 30, isopropanol is recommended as the solvent.

Another, and more preferred way to prepare primary amines of formulas IV, V and VI from ketones of formulas I, II or III (X = O) involves condensing said ketones with the ammonium salt of a lower alkanoic acid and further reducing the imine formed in situ. In addition to the ammonium salts of lower alkanoic acids, other ammonium salts, such as inorganic acids, can also be used.

In practice, solvents of ketone I, II or III (X = O) in a lower alkanol such as methanol are treated with the ammonium salt of an alkanoic acid such as acetic acid and the cooled reaction mixture is treated with a reducing agent, sodium cyanoborohydride. The reaction is allowed to proceed at room temperature for several hours, after which hydrolysis is performed and the product is isolated.

Although one mole of ammonium alkanoate is required per mole of ketone, it is preferable to add an excess thereof to ensure rapid imine formation. An excess as high as ten times can be used without compromising the quality of the final product.

With respect to the amount of reducing agent to be used relative to the molar amount of ketone, it is recommended to use about two moles of sodium cyanoborohydride per mole of ketone.

The reduction reaction time varies from two to three hours at room temperature.

As mentioned above, the preferred solvent is methanol, with ammonium acetate being the preferred ammonium acetate. Isopropanol can also be used as a solvent and is particularly desirable when solvolysis of the 2'-alkanoyl group is to be avoided.

When separating the desired 4 "-deoxy-4" -amino-oleanodomycin derivatives from any non-basic by-products or starting material, the basic nature of the final product 30 is utilized. Thus, the aqueous solution of the product is gradually extracted by raising the pH so that the neutral and non-basic substances are extracted at lower pH values and the product at a pH of about 9. The solvents used for extraction, either ethyl acetate or diethyl ether, are washed with saturated brine and water after extraction. dried over sodium sulfate, the product is obtained by removing the solvent.

6 68059

If further purification is necessary, it can be performed by column chromatography on silica gel according to known methods.

The above-mentioned reductive amination can be performed under other reduction conditions in addition to the use of sodium cyanoborohydride. Some noble metal catalysts, such as a palladium-carbon mixture, can be used effectively with hydrogen and ammonium alkanoate to convert compounds of formulas I, II and III (X = O) to the corresponding compounds of formulas IV, V and 10 VI.

a practically suitable ketone dissolved in a lower alxanol such as methanol or isopropanol is treated with ammonium alkanoate such as ammonium acetate and a 10% palladium on carbon mixture and the resulting suspension is shaken in hydrogen gas at 25-50 ° C, until the theoretical amount of hydrogen has been absorbed.

With respect to the ratios of the amounts of reagents, it is recommended to use a ten-fold excess of ammonium alkanoate-20 to complete the reaction within a reasonable time. The amount of catalyst can vary from ten to fifty percent by weight of the starting ketone. The initial pressure of hydrogen is not critical and it is recommended to use a pressure between one atmosphere and 500 pounds per square inch (35 kg / cm) to reduce the reaction time. Using the above parameters, the reaction time varies from two to six hours.

At the end of the reductive amination reaction, the spent catalyst is filtered off and the filtrate is evaporated to dryness. Purification of the product is carried out as in the above-mentioned method in which sodium cyanoborohydride was used as a reducing agent.

Antibacterial compounds of formula IV wherein R is from one to six carbon atoms

O

35 alkyl, may conveniently be prepared using ketone I (X = O) and the appropriate amine as starting material, and sodium cyanoborohydride as the starting material. To maintain the pH

II

Between 6 68059 and 6-7, an equal molar amount of alkanoic acid, such as acetic acid, is used with the amine. Alternatively, an equivalent amount of hydrogen chloride gas may be used instead of alkanoic acid. The ratio of the amounts of the reagents, the temperature and time of the reaction temperature and the manner of carrying out the reductive amination reaction are the same as the corresponding parameters in the preparation of the compounds in which R 1 is hydrogen, and sodium cyanoborohydride is used as the reducing agent.

Antibacterial compounds of formula IV wherein R 1 is methyl are prepared by alkylation of a reducing agent of formula 4 with 4 "-deoxy-4" -amino-oleandomycin wherein R 1 is hydrogen using formaldehyde and 10% palladium on carbon. carbon mixture.

The ratio of the amounts of the reagents, the reaction temperature, the solvent pressure, the reaction time and the procedure are the same as the parameters in the reaction for preparing the compound IV in which Rg is hydrogen, and hydrogen gas and 10% palladium are used as a reducing agent.

The compounds of the formula I, II or III used as starting materials are prepared from the corresponding compounds of the formula I, II or III in which X = OH and R = alkanoyl having 2 or 3 carbon atoms. The OH group is oxidized to a ketone group as described in FI application No. 780 35 5 (patent 65262).

Removal of the 2'-alkanoyl group is accomplished by a sulfolysis reaction in which the 2'-alkanoyl-4 "-deoxy-4" -oxo-oleandomycin compound is stirred overnight in excess methanol at room temperature. After removal of methanol and purification of the residue, compounds of formulas I, II and III in which R 1 is hydrogen and X is 0 are obtained.

The hydroxyl groups in positions 11 (R = H) and 2 '(R1 = H) of ketones (X = O), I, II and III can be acylated by treating said compounds with two moles of pyridine and an excess of alkane anhydride at ice bath temperature. In practice, the compound containing hydroxyl groups is added to chilled alkane anhydride, followed by the addition of pyridine. After the additions are complete, the ice bath is removed and the mixture is stirred at room temperature overnight. The product is obtained by hydrolyzing the reaction mixture with water and then extracting it with ethyl acetate. Alternatively, the excess alkane anhydride can be removed in vacuo and the residue purified by conventional methods.

Preferred intermediates of formula II or III are 11,2'-diacetyl-4 "-deoxy-4" -oxo-oleandomycin, 11'-acetyl-4 "-deoxy-4" -oxo-oleandomycin, 4 " -deoxy-4 "-oxo-oleandomycin, 2'-acetyl-H" -deoxy-4 "-oxo-oleandomycin, 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-4" -deoxy-4 "-oxo-oleandomycin, 11-acetyl-8,8a-deoxy-8,8a-dihydro-4" -deoxy-4 "-oxo-oleandomycin, 8,8a-deoxy-8,8a -dihydro-4 "-deoxy-4" -oxo-oleandomycin, 2'-acetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin, 11,2'- diacetyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-oleandomycin, 11-acetyl-8, Rain oxy-8,8a-methylene-4 "-deoxy-4 "-oxo-oleandomycin, 8,8a-deoxy-8,8a-methylene-4" -deoxy-4 "-oxo-oleandomycin and 2'-acetyl-8,8a-deoxy-8,8a-methylene- 4 "-deoxy-4" -oxo-oleandomycin.

Oximes of ketones (X = O) are prepared by reacting said ketones with hydroxylamine hydrochloride in methanol-water solution at room temperature. In practice, an excess of hydroxylamine should be used and a triple excess should give a good yield of the desired intermediate. When room temperature and excess hydroxylamine are used in the preparation of the desired oxime derivative, a reaction time of 1-2 hours is obtained. The product is isolated by stirring the reaction mixture in water and basifying to a pH of 9.5, and then extracting it with a water-immiscible solvent such as ethyl acetate.

When O-methylhydroxylamine hydrochloride is used instead of hydroxylamine hydrochloride, reaction 3 68059 results in an O-methyloxime derivative. When O-methylhydroxylamine is used, it is preferable to extend the reaction time to 6-12 hours. The separation of the product is carried out in the same manner as described above for the oxime derivative. O-Acetyloxime compounds (X = N-OCCHg) are prepared by acetylation of the corresponding oxime. Experimentally, one mole of oxime is reacted with one mole of acetic anhydride in the presence of one mole of pyridine. The use of excess anhydride and pyridine speeds up the completion of the reaction and it is recommended to use a two to three times excess. The reaction is preferably carried out in an aprotic hydrocarbon solvent such as benzene or toluene overnight. When the reaction is complete, water is added and the product is separated into a hydrocarbon layer. Alternatively, O-acetyl derivatives can be prepared by treating the required ketone with O-acetylhydroxylamine hydrochloride under the reaction conditions used to prepare oxime derivatives.

Preferred intermediates useful in the preparation of antibacterial 4 "-deoxy-4" -amino-oleanomycin derivatives are 11,2'-diacetyl-4 "-deoxy-4" -oxo-oleandomycinooxime, 11-acetyl-4 "-deoxy-4" -oxo-oleandomycinoxy-25 ml, 11,2'-diacetyl-4 "-deoxy-4" -oxo-oleandomycin-O-acetyloxime, 11-acetyl-4 "-deoxy-4" -oxo -oleandomycin-O-acetyloxime, 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin oxime, 11-acetyl-8,8a deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin-oxyoxime, 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4-" - oxo-oleandomycin-O-acetyloxime, 11-acetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-Olean-domycin-O-acetyloxime, 11,2'- diacetyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-oleandomycin oxime, 35,11-acetyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-oleandomycin oxime, 11,2'-diacetyl-8,8a-deoxy-8,8a-108080 me Tylene-4 "-deoxy-4" -deoxy-4 "-oxo-oleandomycin-O-acetyloxy and 11-acetyl-8,8a-deoxy-8,8a-methylene-4" -deoxy-4 "- oxo-oleandomycin O-asetyylioksiimi.

Among the compounds of the invention, due to their antibacterial utility, the most preferred are 4 "-deoxy-4" -amino-oleandomycin, 11-acetyl-4 "-deoxy-4" -amino-oleandomycin, 11,2'-diacetyl-4 "- deoxy-4 "-amino-oleandomycin, 11-acetyl-8,8a-deoxy-8,8a-dihydro-4" -deoxy-4 "-amino-oleandomycin, 11,2'-diacetyl-10,8,8a -deoxy-8,8a-dihydro-4 "-deoxy-4" -amino-oleandomycin, 11-acetyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -amino-oleandomycin and 11 , 2'-diacetyl-8,8a-deoxy-8,8a-methylene-phenylene-4 "-deoxy-4" -amino-oleandomycin.

To take advantage of the chemotherapeutic effect of the salt-forming compounds of the present invention, it is, of course, preferable to use pharmaceutically acceptable salts. Although water insolubility, high toxicity, or lack of crystalline nature may make a particular salt species unsuitable or less viable for use as such. In some pharmaceutical applications, water insoluble and toxic salts may be converted to the corresponding pharmaceutically acceptable salts as described above, can be converted to any desired pharmaceutically acceptable acid addition salt.

Examples of pharmaceutically acceptable anion-producing acids include hydrochloric, hydrobromic, hydroiodic, nitric and sulfuric and sulfuric acids, as well as phosphoric, acetic, lactic, citric, tartaric, succinic, maleic, gluconic and and aspartic acid.

The stereochemical structure of the starting materials leading to the antibacterial agents of the present invention is the same as that of the naturally occurring agent. The oxidation of the 4 "hydroxyl group to a ketone and the subsequent conversion of said ketone 35 to 4" amines provide an opportunity to deviate the stereochemical structure of the 4 "substituent from the structure of the natural product of 68059. Thus, when compounds I, II and III (X = 0) If two isolated epimeric amines are present in the final product in varying proportions depending on the choice of synthetic method, it can be found that two epimeric amines are present in the final product in varying proportions depending on the choice of synthetic method. repeatedly from a suitable solvent 10 until a constant melting point is reached.Another epimer that was present to a small extent in the actual isolated solid product is the predominant product in the mother liquor, which can be recovered therefrom by methods known to those skilled in the art, such as evaporation. the mother liquor to dryness and recrystallizing the residue several times until a constant melting point is reached for the product, or using chromatographic methods.

Although epimers can be separated from said mixture by methods known to those skilled in the art, for practical reasons it is preferable to use said mixture as such isolated from the reaction. However, it is usually preferable to purify the epimer mixture by recrystallization at least once from a suitable solvent, to carry out the purification by column chromatography, by solvent demonstration or by trituration from a suitable solvent. Said purification, although not separating the epimers, removes such additional substances as starting materials and undesired by-products.

30 The absolute stereochemical representation of the epimers has not been elucidated. However, both epimers of a particular compound have the same type of activity, e.g. as antimicrobial agents.

The following tables show the antibacterial properties of the compounds of the invention compared to the corresponding properties of oleandomycin. The results are expressed as MIC values (the lowest possible concentration of test compound that inhibits the growth of the microorganism in question). Table I shows the results for compounds of formula IV which contain a group 1-4 to 0 in position _ch2 5 8 (8a), while Table II shows the results for compounds of formula V which contain a group -CH2 in position 8 (8a). Table III depicts the results for compounds of formula VI containing a CH 2 group at position 8 (8a), and Table IV depicts the results for the reference compound, oleandomycin. The results show that the antibacterial activity of the compounds of the present invention, especially against organisms Nos. 13, 14, 15 and 24, is considerably better than that of the reference compound.

Il 68059

TABLE I

MIC, μg / ml.

A compound of formula IV wherein = H

Microorganism ^ ^ = H ^ = CH3CO

R = CH3CO R = H R = H

(1) Staph, aur. 01A005 1.56 3.12 6.25 (2) "" 01AO 52 3, 1 2 3.1 2 6.25 (3) "" 01A109 R> 200 200> 50 (4) "" 01A110> 200 200> 50 (5) "" 01A111 R 0,78 0,1 0 0,78 (6) "" 01A087 RR> 200 100> 50 (7) "" 01A400 R 1,56 3,12 6,25 (8) Strp . Fae. 02A006 1.56 3.12 3.12 (9) Strp. pyog. 02C203 0.78> 0.10 1.56 (10) "" 02C020 R> 200 -> 50 (11) Myco. smeg. 05A001 6,25 25> 50 (12) B. sub. 06A001 1.56 3.12 3.12 (13) E. coli 51A229 12.5 50 50 (14) "" 51A266 25 25 50 (15) "" 51A125 R 25 100> 50 (16) Ps. Aerug. 52A104> 200 100> 50 (17) Klebs. pn. 53A009 25 100> 50 (18) "" 53A031 R 50 100> 50 (19) Prot. mira. 57C064> 200> 200> 50 (20) Prot. Morg. 57G001> 200 100> 50 (21) Salm. chol-Sun. 58B242 50 100 (22) Ps. typhm. 58D009 50 100> 50 (23) "" 58D013-C 25 100 (24) Past.multo. 59A001 1.56 6.25 6.25 (25) Serr. Mar. 63A017> 200 100> 50 (26) Ent. Aero. 67A040 50 -> 50 (27) Ent.eloa 67B003 100 -> 50 (28) Neiss. sic. 66C000 _ <0.10 <0.10 j ι4 68059

TABLE II

MIC, μg / ml.

Compound of formula V Microorganism R 1 = H = H

R = H R = CH3CO

(1) Staph, aur. 01A005 6,25 3,12 (2) "" 01A052 6,25 3,12 (3) "" 01A109 R> 200> 200 (4) "" 01A110 R> 200> 200 (5) "" 01A111 R 0, 78 0.78 (6) "" 01A087 RR> 200> 200 (7) "" 01A400 R 6.25 3.12 (8) Strp. fae. 02A006 3.12 3.12 (9) Strp. pyog. 02C203 0.20 0.20 10) "" 02C020 R 50 200 11) Myco. smeg. 05A001 6.25 12.5 12) B. sub. 06A001 6,25 3,12 13) E. coli 51A229 100 25 14) "" 51A266 100 50 15) "" 51A125 R 100 50 16) Ps. Aerug. 52A104> 200> 200 17) Klebs. pn. 53A009 200 100 18) "" 53A031 R> 200 100 19) Prot. mira. 57C064> 200> 200 20) Prot. Morg. 57G001 200> 200 21) Salm. Chol-Sun. 58B242 200 100 22) Ps. typhm. 58D009 200 50 23) "" 58D013-C 3.12 100, 24) Past.multo. 59A001> 200 3.12 25) Serr. November. 63A017> 200> 200 26) Ent.aero. 67A040> 200 100 27) Ent.cloa. 67B003 100 200 28) Neiss. sic. 66C000 0.20 £ 0.10 teaspoon is 68059

TABLE III

MIC, μg / ml.

Compound of formula VI Microorganism = H

R = CH3CO

(1) Staph, aur. 01A005 0,78 (2) "" 01A052 1,56 (3) "" 01A109 R> 200 (4) "" 01A110 R> 200 (5) "" 01AI 11 R 0,78 (6) "" 01A087 RR> 200 (7) "" 01A400 R 1.56 (8) Strp. gar. 02A006 1.56 (9) Strp. pyog. 02C203 0.20 (10) "" 02C020 R> 200 (11) Myco. smeg. 05A001 (12) B. sub. 06A001 0.78 (13) E. Coli 51A229 25 (14) "" 51A266 50 (15) "" 51A125 R 50 (16) Ps. Aerug. 52A104> 200 (17) Klebs. pn. 53A009 100 (18) "" 53A031 R 50 (19) Prot. mira. 57C064> 200 (20) Prot. Morg. 57G001> 200 (21) Salm. chol-Sun. 58B242 100 (22) Ps. typhm. 58D009 25 (23) "" 58D013-C 25 (24) Past, multo. 59A001 3.12 (25) Serr. November. 63A017> 200 (26) Ent. Adro. 67A040 (27) Ent.eloa. 67A003 100 (28) Neiss. sic. 66C000 £ 0.10 16 68059

TABLE IV

MIC, μg / ml

Microorganism Oleandomycin (1) Staph, aur. 01A005 0,78 (2) "" 01A052 1,56 (3) "" 01A109 R 100 (4) "" 01A110 100 (5) "" 01 AI 11 R 0,78 (6) "" 01A087 RR 100 (7) ) "" 01A400 R 1,56 (8) Strp. fae. 02A006 1.56 (9) Strp. pyog. 02C203 0.78

(10) "" 02C020 R

(11) Myco. smeg. 05A001 100 (12) B. sub. 06A001 0.78 (13) E. coli 51A229 100 (14) "" 51A 2 6 6 1 00 (15) "" 51A125 R 100 (16) Ps. Aerug. 52A104 100 (17) Klebs. pn. 53A009 100 (18) "" 53A031 R 100 (19) Prot. mira. 57C064 (20) Prot.morg. 57G001 100 (21) Salm. chol-Sun. 58B242 (22) Ps. typhm. 58D009 100

(23) "" 58D013-C

(24) Past, multo. 59A001 12.5 (25) Serr. November. 63A017 100 (26) Ent. aero. 67A040 100 (27) Ent. life. 67B003 100 (28) Neiss. sic. 66C000 12.5 17 68059

For external use, the compounds of the invention are preferably combined with a pharmaceutically acceptable carrier, such as a vegetable or mineral oil or skin emollient. Similarly, it may be dissolved or dispersed in liquid carriers or solvents such as water, alcohol, glycols, or mixtures thereof, or other pharmaceutically acceptable inert media; that is, to media that do not adversely affect the active ingredient-10 moiety. For such purposes, it is generally accepted to use active ingredient concentrations of 0.01 to 10% by weight of the total preparation.

For internal use, these new compounds may be administered orally or parenterally, e.g., by subcutaneous or intramuscular injection, at a dose of 1 to 200 mg per kilogram of body weight per day. The preferred dosage range is about 5 to 100 mg per kilogram of body weight per day. Suitable carriers for parenteral injection may be aqueous, such as water, isotonic saline, isotonic dextrose, Ringer's solution, or non-aqueous, such as vegetable fatty oils (cottonseed, peanut, corn, sesame oil), dimethyl sulfoxide and other aqueous sulfoxide. carriers that do not affect the therapeutic efficacy of the preparation and are not toxic in the volumes or proportions used (glycerol, propylene glycol, sorbitol). It is also possible to make preparations which are suitable for the preparation of a solution just before dosing. Such preparations may include liquid diluents, for example, propylene glycol, diethyl carbonate, glycerol, sorbitol, etc .; buffering agents, hyaluronidase, topical anesthetics and inorganic salts to provide the desired pharmacological properties. These compounds may also be combined with a variety of pharmaceutically acceptable inert carriers, including solid diluents, aqueous media, non-toxic organic solvents in the form of capsules, round and conical tablets, lozenges, dry mixtures, suspensions, solutions, elixirs and excipients. suspensions. In general, Compounds-10 are used in a variety of dosage forms in concentrations ranging from 0.5 to about 90% by weight of the total preparation.

The following examples illustrate the invention.

Example 1 2'-Acetyl-N-deoxy-4 "-oxo-oleandomycin Dimethyl sulfide (0.337 ml) is added to a cloudy solution of 467 mg of N-chlorosuccinimide in 20 ml of toluene and 6 ml of benzene which has been cooled - After stirring at 0 [deg.] C. for 20 minutes, the mixture is cooled to -25 [deg.] C. and 1.46 g of 2'-acetyl-oleandomycin and 15 ml of toluene are added. Stirring is continued for two hours at -20 [deg.] C., after which 0.46 ml of triethylamine are added, the reaction mixture is kept at -20 [deg.] C. for a further five minutes and then allowed to warm to 0 [deg.] C.

The mixture is poured with stirring into 50 ml of water and 50 ml of ethyl acetate. The pH of the aqueous mixture is adjusted to 9.5 by adding aqueous sodium hydroxide solution. The organic layer is then separated, dried over sodium sulfate and concentrated in vacuo to a white foam (1.5 g). Trituration with diethyl ether gives 864 mg of crude product which, recrystallized twice from methylene chloride-diethyl ether gives 212 mg of pure product m.p. 183 to 185.5 ° C.

il ι9 68059

Analysis Calculated for C 61 H 8 8.5 N 1.9 Found: C 60.9 H 8.4 N 1.9 NMR (4, CDCl 3): 5.60 (1H) m , 3.50 (3H) s, 2.73 (2H) m, 2.23 (6H) s and 2.03 (3H) s.

Following the procedure described above and using 2'-propionyl oleandomycin as starting material, 2'-propionyl-4 "-deoxy-4" -oxo-oleandomycin is obtained.

Example 2 4 "-deoxy-4" -oxo-oleandomycin

A solution of 1.0 g of 2'-acetyl-4 "-deoxy-4" -oxo-oleandomycin in 20 ml of methanol is stirred at room temperature overnight. The solution is concentrated in vacuo to give the desired product as a white foam, 15,937 mg.

NMR (δ, CDCl 3): 5.60 (1H) m, 3.50 (3H) s, 2.85 (2H) m and 2.26 (6H) s.

Example 3 11,2'-Diacetyl-4 "-deoxy-411-oxo-oleandomycin 20 A. By acetylation in 4.0 ml of 727 mg of 2'-acetyl-4" -deoxy-4 "-oxo-oleandomycin is added in an ice bath. Acetic anhydride cooled to 0 [deg.] C. in the presence of nitrogen gas, after 5 minutes 0.158 ml of pyridine are added and the resulting cloudy suspension is stirred at room temperature overnight, the resulting solution is stirred in a layer of ethyl acetate and the pH is adjusted to 7.2 by adding solid sodium bicarbonate and then to 9.5 using 1N sodium hydroxide solution.

The organic layer was separated, washed successively with water and saturated brine, and dried over sodium sulfate. Removal of the solvent under reduced pressure gives 588 mg of the desired product.

NMR (cf, CDCl 3): 3.48 (3H) s, 2.63 (2H) m, 2.26 (6H) s and δ 2.06 (6H) s.

20 6 3 0 5 9

By repeating this procedure and using the required 2'-alkanoyl-4 "-deoxy-4" -oxo-oleandomycin and acylating agent, the following compounds can be prepared: 11-acetyl-2H-propionyl-4 "-deoxy-4" -oxo- oleandomycin, 11-pro-5-pionyl-21-acetyl-4 "-deoxy-4M-oxo-oleandomycin and 11,2'-dipropionyl-4" -deoxy-4 "-ox-oleandomycin.

B. By oxidation

Add 3.36 ml of dimethyl sulfide to a dry flask cooled to -5 ° C with a magnetic stirrer and nitrogen inlets containing 4.5 g of N-chlorosuccinimide, 50 ml of benzene and 150 ml of toluene. After stirring at 0 [deg.] C. for 20 minutes, the contents of the flask are cooled to -25 [deg.] C. and 5.0 g of 11,2-di-acetyl-oleandomycin in 100 ml of toluene are added. Cooling and stirring are continued for 2 hours, after which 4.73 ml of triethylamine are added. The reaction mixture is stirred at 0 ° C for 15 minutes, then poured into 500 ml of water. The pH is adjusted to 9.5 with 1N aqueous sodium hydroxide solution and the organic layer is separated, washed with water and saturated brine, and dried over sodium sulfate. Removal of the solvent in vacuo gives 4.9 g of the desired product, identical to that prepared in Example 3A, as a foam.

NMR (δ, CDCl 3): 3.48 (3H) s, 2.61 (2H) m, 2.23 (6H) s and 2.03 (6H) s.

Example 4 11-Acetyl-4,11-deoxy-4 "-oxo-oleandomycin

A solution of 4.0 g of 11,21-diacetyl-4 "-deoxy-4" -oxo-oleandomycin in 75 ml of methanol is stirred at room temperature overnight. The reaction mixture is concentrated under reduced pressure to give a foamy product. This residue dissolved in diethyl ether and treated with hexane gave 2.6 as a white solid, m.p. 112-117 ° C.

NMR (δ, CDCl 3): 3.43 (3H) s, 2.60 (2H) m, 2.23 (6H) s and 2.01 (3H) s.

1) 21 68059

Similarly, using 11,2'-propionyl-4 "-deoxy-4" -oxo-oleandomycin or 11-propionyl-2'-acetyl-4 "-deoxy-4" -oxo-oleandomycin, 11-propionyl- 4 "-deoxy-4" -oxo-oleandomycin i ini.

Example 5 11,2'-Diacetyl-4 "-deoxy-4" -oxo-oleandomycin

The reaction mixture comprising 1.0 g of 11,2'-diacetyl-Olean domycin, 7.09 ml of dimethyl sulfoxide and 9.44 ml of acetic anhydride is stirred at room temperature for four days. The resulting yellow solution is mixed with water to which benzene has been added as a layer. The pH is then adjusted to 9.5 with 1N aqueous sodium hydroxide solution and the organic layer is separated, dried over sodium sulfate and evaporated to dryness in vacuo. The resulting yellow oily foam 15 is purified by chromatography on 20 g of silica gel using a 9: 1 mixture of chloroform and acetone as eluent. Removal of the solvent from the fractions gives 800 mg of the desired product and 110 mg of by-product.

Similarly, the use of 11-acetyl-2'-propionylole-20 andomycin as 11,2'-dipropionyl oleandomycin or 11-propionyl-2'-acetyl oleandomycin as a starting material in the oxidation process described above gives 11-acetyl-2'-pro pionyl-4 "-deoxy-4" -oxo-oleandomycin, 11,2'-dipropionyl-4-deoxy-4 "-oxo-oleandomycin and 11-propionyl-21-acetyl-4-deoxy-4" -oxo -oleandomysiini.

Example 6 2'-Acetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4''-oxo-oleandomycin

Add 6.0 ml of dimethyl sulphide to a dry flask cooled to -5 ° C with a magnetic stirrer and a nitrogen inlet containing 11.6 g of N-chlorosuccinimide, 750 ml of toluene and 250 ml of benzene and stir the solution thus obtained. minutes. The temperature is further lowered to -20 ° C and 25 g of 2'-acetyl-8,8a-deoxy-8,8a-dihydro-35-oleandomycin in 500 ml of toluene are added. After stirring for 2 hours at -20 ° C, 11.4 ml of triethylamine are added and the reaction mixture is gradually allowed to warm to 0 ° C. It is then poured into 1880 ml of 1500 ml of water and the pH is adjusted to 9.5 with 1N sodium hydroxide solution. The organic layer is separated, washed successively with water (3x) and brine (1x) and dried over sodium sulfate. Removal of the solvent under reduced pressure gives a foam which, recrystallized from diethyl ether, gives 13 g of pure product, m.p. 197-199 ° C.

NMR (/ ', CDCl 3): 5.11 (1H) m, 3.51 (3H) s, 2.25 (6H) s and 2.03 (3H) s.

Example 7 PO 8,8a-Deoxy-8,8a-dihydro-4 "-deoxy-N-oxo-oleandomycin

A suspension of 2.0 g of 2'-acetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin in 100 ml of methanol is stirred at room temperature for 20 hours. The reaction mixture is concentrated under reduced pressure to give 1.8 g of the desired product as a white foam.

NMR (δ, CDCl 3): 5.30 (1H) m, 3.51 (3H) s and 2.26 (6H) s.

Example 8 11,2 * -Diacetyl-8,8a-deoxy-8,8a-dihydro-α, 1-deoxy-U, 1-oxo-oleandomycin 20. ...

2.8 ml of pyndin are added to an ice-bath-cooled suspension of 13.0 g of 2'-acetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin in 65 ml of acetic anhydride. The bath is then removed and the reaction mixture is stirred at room temperature overnight. The resulting solution is then added to 500 mL of all water and 300 mL of ethyl acetate. The pH is adjusted to 7.0 with solid sodium bicarbonate and then 9.5 with 4N aqueous sodium hydroxide solution. The organic layer is separated, washed with water (2x) and brine (1x) and dried over sodium sulfate. Removal of the solvent in vacuo gives the crude product as a foam.

Since the chromatographic results of a test batch of the crude product show that the reaction is not complete, the crude foam is mixed with 28 any pyridine and 79 any acetic anhydride and stirred at room temperature for 72 hours. The reaction mixture is worked up as above to give 12.4 g of the desired product.

NMR (δ, CDCl 3): 3.51 (3H) s, 2.26 (6H) s and 2.10 (6H) s.

23

Example 9 68059 11-Propionyl-21-acetyl-8,8a-deoxy-8,8a-dihydro-4n-deoxy-U1, oxo-oleandomycin

The procedure of Example 8 is repeated using 6.5 g of 2'-acetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleo-andomycin, 40 ml of propionic anhydride and 14 ml of pyridine as starting materials. After a reaction time of 72 hours, the mixture is worked up as described in Example 8 to obtain the desired product.

Example 10 11-Acetyl-8,8a-deoxy-8,8a-dihydro-4,11-deoxy-4'1-oxo-Olean domycin

A solution of 11.5 g of 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin in 100 ml of methanol is stirred at room temperature overnight. Evaporation of the reaction mixture to dryness under reduced pressure gives 10.6 g of a crude foamy product. The crude product is dissolved in chloroform and the solution is applied to a silica gel column. After 3 L of chloroform has passed through the column, the product is initialized with chloroform / methanol (19: 1). Eight hundred-drop fractions are pooled with an automatic fraction collector. Fractions 50-56, 57-62, 63-69 and 70-80 are combined and evaporated to dryness in vacuo to give 2.9 g of pure product.

NMR (δ, CDCl 3): 3.55 (3H) s, 2.31 (6H) s and 2.05 (3H) s.

Example 11 11-Propionyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4'-oxo-oleandomycin Starting from Example 11, 11-propionyl-2'-acetyl-8,8a-deoxy-8 , 8α-dihydro-4 "-deoxy-4 '' - oxo-oleandomycin and following the procedure of Example 10 the desired product is prepared.

Example 12 21-Acetyl-8,8a-deoxy-8,8a-dihydro-oleancmycin To a solution of 5.0 g of 8,8a-deoxy-8,8a-dihydro-oleandomycin in 15 ml of benzene is added 0, 73 ml of acetic anhydride and the reaction mixture thus obtained are stirred at room temperature for 1.5 to 35 hours. The solution is poured into 100 ml of water and the pH is adjusted to 7.5 with 24 68059 solid sodium bicarbonate and then to 9.5 with 1N aqueous sodium hydroxide solution. After stirring for 10 minutes, the organic layer is separated and washed successively with water (2x) and saturated brine (1x) and then dried over sodium sulfate. Removal of the solvent under reduced pressure gives 4.9 g of the desired product, m.p. 202-204 ° C.

NMR (δ, CDCl 3): 5.05 (1H) m, 3.40 (3H) s, 2.25 (6H) s and 2.05 (3H) s.

Similarly, by replacing acetic anhydride with an equivalent amount of propionic anhydride, 2'-propionyl-8,8a-de-10-oxy-8,8a-dihydro-oleandomycin is prepared.

Example 13 21-Propionyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin

To a solution of 375 ml of toluene and 125 ml of Bent-15 fungus is added 5.8 g of N-chlorosuccinimide, and the mixture is stirred at room temperature for 15 minutes. The reaction mixture is cooled to -5 ° C and 3.0 ml of dimethyl sulfide are added and stirring is continued for a further 20 minutes. The temperature is reduced to -20 [deg.] C., after which 12.8 g of 21-propionyl-8,8a-deoxy-8,8a-di-20-hydro-oleandomycin in 250 ml of toluene are added. After 2 hours, 5.7 ml of diethylamine are added and the cooling bath is removed. When the reaction temperature reaches 0 ° C, the mixture is poured into 750 ml of water. The pH is adjusted to 9.5 with 1N sodium hydroxide solution and the organic layer is separated. The organic phase washed with water (3x) and brine is dried and concentrated under reduced pressure to give the desired product.

Example 14 11,2,2-Diacetyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy-α" -oxo-oleandomycin 30-

To a cloudy solution of 434 mg of N-chlorosuccinimide in 15 ml of toluene and 5 ml of benzene cooled to -5 ° C is added 0.327 ml of dimethyl sulfide. After stirring for 20 minutes at 0 DEG C., the reaction mixture is cooled to -25 DEG C. and 500 mg of 11,2'-diacetyl-8,8a-deoxy-8,8a-methylene oleandomycin and 10 ml of toluene are added. Stirring is continued for two

II

25809 hours at -20 [deg.] C. and then 0.46 ml of triethylamine and 1 ml of toluene are added. The cooling bath is removed and the reaction mixture is allowed to warm to 0 ° C. It is then poured into 50 ml of water and 50 ml of ethyl acetate. The pH is carefully adjusted to 9.5 and the organic layer is separated, dried and evaporated to dryness. In this way, 500 mg of the desired product are obtained in the form of a slightly moist white foam.

NMR (cis, CDCl 3): 3.50 (3H) s, 2.30 (6H) s, 2.06 (6H) s and 0.58 (4H) m.

Example 15 Using the appropriate 8,8a-deoxy-8,8a-methylene oleandomycin as starting material and following the procedure of Example 14, the following 8,8a-deoxy-8,8a-methylene-15 4 "-deoxy-4" -oxo-oleandomycin derivatives are prepared : .AT “,, s Λ τχ 20 R0

0 Y

11 ΥΥ'ο ° 0CH ~ 25 3 - h 0 0 CH.C- CH0CH0C- 30 J 61 Q 0 0 ch3ch2c- ch3c- 0 0 CH3CH2C- CH3CH2C- 0 3 5 "H- CH.C- ά 0 H- CH3CH2C - 26 68059

Example 16 11-Acetyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy- * 4" -oxo-oleandomycin * 400 mg of 11,2'-diacetyl-8,8a-deoxy-8,8a a solution of methylene-5 4 "-deoxy-4" -oxo-oleandomycin in 10 ml of methanol, after stirring at room temperature overnight, is concentrated in vacuo to give 270 mg of the desired product.

NMR (δ, CDCl 3): 3.46 (3H) s, 2.26 (6H) s, 2.03 (3H) s and 0.56 (4H) m.

Example 17

By repeating the procedure of Example 16 and starting from the appropriate 2'-alkanoyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-oleandomycin, the following compounds are obtained:. r, «“ a’j r \ RO »-» / \

20 .1 L

"\> fY i Y" ° 0CH3 25 Starting material: Product R ^ R ^ 0 0 0 CH3C "CH3CH2C- ch3c ~ H- 0 0 0 ch3ch2c- ch3c- ch3ch2c- H- 0 0 0 35 CH3CH2C- CH3CH2C- CH3CH2C H- H- CH C- H- H- d 0 H- CH3CH2C- H- H- 27 68059

Example 18 11,2'-Diacetyl-8,8a-deoxy-8α-methylene-N, N-deoxy-Htl-oxo-oleandomycin

18 ml of methylene dichloride and 1.97 ml of dimethyl sulfoxide are placed in a dry flask equipped with a nitrogen inlet and the solution thus obtained is cooled to -60 ° C. Trifluoroacetic anhydride (3.9 ml) is gradually added and stirring is continued in the cold for 10 minutes. The reaction mixture is further cooled to -70 [deg.] C. and 5.34 g of 11,2'-diacetyl-8,8a-deoxy-8,8a-methylenea oleandomycin dissolved in 27 ml of methylene dichloride are added dropwise so that the temperature does not rise above -50 ° C. The reaction mixture is then cooled back to -70 ° C, 9.69 ml of triethylamine 15 are added and the mixture is kept at -70 ° C for a further 10 minutes. The reaction mixture is allowed to warm to -10 ° C and then poured into 7 ml of water. Adjust the pH to 9.5 with 1N sodium hydroxide solution and separate the methylene dichloride. The organic layer is then washed with water (2 x 30) and brine (1 x 20) and dried over sodium sulfate. Removal of the solvent in vacuo gives 6.2 g of product which, except for a minor impurity, is identical to the product of Example 14.

Example 19 By repeating the procedure of Example 18 and starting from the required 2'-alkanoyl-8,8a-deoxy-8,8a-methylenoleandomycin, the following 4 "-deoxy-4" -oxo derivatives are obtained:

30 .X

Vix

o I

0CH3 __ TT— 28 6 8 0 5 9 R R1 0 CH3CH2C- CH3 ^ '

5 O

H- CH ~ C- 0 H- CH3CH2C-

0 O

CH3CH2S CH3CH2C- CH3CH2C-

Example 20 11,2'-Diacetyl-N-deoxy-N-oxo-oleandomycin oxime

To a solution of 18.1 hydroxylamine hydrochloride in 300 ml of water and 200 ml of methanol is added 50 g of 11,2'-diacetyl-4 "-deoxy-4" -oxo-oleandomycin and the reaction mixture is stirred at room temperature for one hour. . The resulting solution is added to water and the pH is then adjusted to 7.5 with solid sodium bicarbonate and then to 9.5 g of 1N sodium hydroxide. The product is extracted into ethyl acetate and the dried extracts are concentrated to about 25,170 ml. Hexane is added to the heated ethyl acetate solution to the cloud point and the cloudy solution is cooled. The precipitated product is filtered off and dried, 29.8 g, m.p. 2 2 3, 5-22 5 ° C.

NMR (δ, CDCl 3): 3.30 (3H) s, 2.65 (2H) m, 2.35 (6H) s and 2.10 (3H) s.

In a similar manner, starting from O-acetylhydroxylamine hydrochloride and the required ketone and following the above procedure, 2'-acetyl-4 · deoxy-4 "-oxo-oleandomycin-O-acetyloxime is obtained.

35 Example 21

By repeating the procedure of Example 20 and starting with the appropriate 1 + "- deoxy - * +" - oxo-oleandomycin, the following oximes are obtained: 29 68059 "(CH3> 2

° ^ rJ $ v, A

RQ, (llli>; VtP- Ό "^

>, <V 0 Y

N ΌΜ /, Τ T

l | γ ^ N-OH

ο och3 10 R R] _ 0 15 H- CH3C- 2 H- ch3ch2 £ -

0 M

CH3C- ch3ch2c- 20 O o CH3CH2C- CH3 ^ "0 n CH3CH2C- ch3ch2c- 25 ° CH3C- h" 0 CH3CH2C- h- H- H- 30

Example 22 11-Acetyl-U "-deoxy-1 * M-oxo-oleandomycin oxime 500 mg of nJ2'-diacetyl-1'-deoxy-4, '- oxo-oleandomycin oxime in 100 ml of a methanol-containing solution se-35 is stirred for 72 hours at room temperature and then evaporated to dryness under reduced pressure The foam thus obtained is recrystallized from ethyl acetate-hexane, 372 mg, mp 184-186 ° C.

NMR (X, CDCl 3): 3.30 (3H) s, 2.66 (2H) m, 2.36 (6H) s and 2.10 (3H) s.

Example 23 Using the required oxime as starting material and following the procedure of Example 22, the following compounds are prepared: 10 R1

o xA

"T I I

ro, A xAo / \ 15 x x '0l \' T "

“Vll-OH

o! 0CH3 20 Starting material Product R Rx R R1 0 H- CH C- H- H- 2 6 ^ ^ 0 H- CH CH C- H- H- 0 d Z0 0 ch3c- ch3ch2c- ch3c- h- 0 0 ° 30 CH3CH2C - ch3c- ch3ch2c- h- 0 0 0 CH3CH2C- CH3CH2C- CH3CH2C- h- f tl 68059

Example 24 11,2 * -Diacetyl-4 "-deoxy-4" -oxo-oleandomycin-O-acetyloxime

To a cloudy solution of 20 g of 11,2'-diacetyl-4 "-deoxy-4" -xo-5-oleandomycin oxime in 250 ml of benzene is added, with stirring, 8.21 ml of pyridine and then 9.62 ml of acetic anhydride, and thus the resulting reaction mixture is stirred at room temperature overnight. The solution is poured into water and the organic layer, washed successively with water 10 and saturated brine and dried, is evaporated to dryness. The foamy residue recrystallized from ethyl acetate-hexane gave 13.4 g of pure product, m.p. 198-202 ° C.

NMR (δ, CDCl 3): 3.38 (3H) s, 2.66 (2H) m, 2.33 (6H) s, 2.26 (3H) s and 2.10 (6H) s.

Example 25

By repeating the procedure of Example 24 using the appropriate oxime as starting material, the following products are obtained: 20 N (CH 2) 0

dX I

° \ / \ ^ u 1 25 R0 // (/ s o ^ 0

VVx S

0 Y N-O-CCH3 30 - 0CH3 H 32 K, 68059 O ό

Il ·· 5 CH3C- CH3CH2C- O · o

Il II

CH3CH2C ~ CH3C- 0 CH3C- h- 10 o

II

ch3ch2c- H-

Example 26 To 11 ml of water and to 50 ml of methanol are added 1.25 g of methoxyamine hydrochloride and 2, 11,2'-diaethyl-U'1-deoxy-III-oxo-oleandomycin-O-methyloxime. 5 g of 11,2'-diacetyl-4 "-deoxy-4-oxo-oleandomycin and the resulting reaction mixture are stirred at room temperature overnight. The solution is stirred in water and the pH is adjusted to 7.5 with solid sodium bicarbonate and then to 9.5 with 1- The product is extracted with ethyl acetate and the extract is dried over sodium sulfate, removing the solvent in vacuo to give 2.4 g of the desired product as a white foam.

NMR (δ f, CDCl 3): 3.88 (3H) s, 3.26 (3H) s, 2.56 (2H) m, 2.30 (6H) s and 2.06 (6H) s.

Example 27

Following the procedure of Example 26, starting from methoxyamine hydrochloride and the required 4 "-deoxy-4" -oxo-oleandomycin, the following compounds are prepared: 2'-acetyl-4 "-deoxy-4 '-oxo-oleandomycin O-methyloxime, 2'-propionyl-4 "-deoxy-4" -oxo-oleandomycin-O-methyloxime, 11-acetyl-2'-propionyl-4 "-deoxy-4" -oxo-oleandomycin-O- methyloxime, 35-11-propionyl-2'-acetyl-4 "-deoxy-4" -oxo-oleandomycin-O-methyl-oxime, 11-acetyl-4 "-deoxy-4" -oxo-oleandomycin-O- methyl oxime and 11-propionyl-4 "-deoxy-4" -oxo-oleandomycin-O-methyloxime.

ti 33 68059

Example 28 11,21-Diacetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin oxime

A solution of 49.0 g of 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin and 18.1 g of hydroxylamine hydrochloride in 300 ml water and 300 ml of methanol, stirred at room temperature for 1.5 hours. The resulting solution is mixed with 250 ml of water and the pH is adjusted to 7.5 and 9.5 with solid sodium bicarbonate and 1N sodium hydroxide solution, respectively. The product is extracted with ethyl acetate, which is then dried over sodium sulfate and evaporated to dryness. The residue, recrystallized from ethyl acetate-hexane, gives the desired product.

15 Example 29

Following the procedure of Example 28 and starting from the appropriate 4 "-deoxy-4" -oxo-oleandomycin and the necessary hydroxylamine derivatives, the following compounds are prepared: 20 [mu] l (ci \ χ

O

H- CH 2 Cl 2 -N-OH

O O

C «3CH2C- CH3C- N-OH

O

<1

ai3C- H- N-OH

O

10 CH 3 CH 2 E - H- N-OH

O

H- CH3CH2C- N-OH

O 0

^ 3 ^ 2 ° "CH3C" N-OCH

15 O «« CB3C- ch3c “N-OCH3

O

II

H-CH3C- N-OCH3

O

20 "ch3c- n- n-och3

O

CH 3 CH 2 C-h-N-OCH 3 Example 30 11, 2'-Diacetyl-8,8a-deoxy-8,8a-dihydro-11-deoxy-11-oxo-oleandomycin-O-acetyloxime

To a solution of 9.9 g of 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin oxime and 4.1 ml of pyridine in 125 ml: in benzene, 4.81 ml of acetic anhydride are added and the resulting reaction mixture is stirred at room temperature overnight. The reaction mixture is poured into water and the pH is adjusted to 7.5 and 9.5 with solid sodium bicarbonate and 1N sodium hydroxide, respectively. The benzene layer is separated, dried over sodium sulfate and concentrated to give the desired product as a white foam.

il 35 68059

Example 31

By repeating the procedure of Example 30 starting from the appropriate oxime, the following compounds are obtained: 2'-acetyl-8,8a-deoxy-8,8a-dihydro-4n-deoxy-4 "-oxo-oleandomycin-O-ace-5-style oxime, 11- propionyl-2'-acetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin-O-acetyloxime, 11-propionyl-8,8a-deoxy-8,8a -dihydro-4 "-deoxy-4" -oxo-oleandomycin-O-acetyloxime, 2'-propionyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin- O-acetyloxime and 11-acetyl-8,8a-10 deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin-O-acetyloxime.

Example 32 11-Propionyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin oxime.

A solution of 500 mg of 11-propionyl-2'-acetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin oxime in 100 ml of methanol is stirred at room temperature overnight. The solution is evaporated to dryness and the foamy residue is purified by recrystallization from ethyl acetate-hexane.

In a similar manner, 8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin oxime is prepared from 2'-acetyl-8,8a-deoxy-8,8a-dihydro-4 "-deoxy- 4 "-oxo-oleandomysiinioksiimista.

Example 33 11,2'-Diacetyl-8,8a-deoxy-8,8a-methylene-4,1-deoxy-4'-oxo-oleandomycin-O-methyloxime

A solution of 90 mg of 11,2'-diacetyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-oleandomycin and 45 mg of methoxyamine hydrochloride in 2 ml of water and 2 in methanol, stirred at room temperature overnight. The solution is poured into water and the pH is adjusted to 7.5 and 9.5 with solid sodium bicarbonate and 1N sodium hydroxide, respectively. The product is extracted with ethyl acetate and the organic layer is dried and concentrated to give 89.2 mg of the desired product.

NMR (δ, CDCl 3): 5.56 (3H) s, 3.33 (1.5H) s, 3.26 (1.5H) s, 2.28 (6H) s, δ 2.06 (6H ) s and 0.56 (4H) m.

36 68059

Example 34 11,2'-Diacetyl-8,8a-deoxy-8α-methylene-1'-deoxy-411-oxo-oleandomycin

By repeating the procedure of Example 33 using 10.0 g of 11,2'-diacetyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-oleandomycin, 3.4 g of hydroxylamine hydrochloride, 50 ml of methanol and 50 ml of water, 9.2 g of the desired product are obtained, which can be further purified by recrystallization from ethyl acetate, m.p. 17 7-18 ° C.

NMR (4, CDCl 3): 3.35 (1.5H) s, 3.25 (1.5H) s, 2.33 (6H) s, 2.06 (6H) s and 0.53 (4H) s. m.

Example 35

Following the procedure of Example 33, using the appropriate hydroxylamine hydrochloride and the corresponding 8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-Olean domycin, the following compounds are prepared: R1X »'“ ah

r> X JL

V <'"[[RO X o / 25 fxjt 30 0CH3 ti 37 68059

R R X

CH3C-CH3CH2C- n-oh 0 0

5 CH3CH2C- ch3C- N-OH

0 0 CH3CH2C- CH3CH2C- n-oh 0

ln H- CH.C- N-OH

d 0 H- ch3ch2c- n-oh 0

CH c- H- N-OH

15 0 0 CH3CH2C-CH3C "N-OCH3 0 0 CH3CH2C- CH3CH2C- N-OCH3 0 20 H-CH3C- N-OCH3 0 CH3C- h-N-OCH3 2 5 Example 3 6 11,21-diacetyl-8,8a -deoxy-8α-methylene-4'-deoxy-4'1-oxo-oleandomycin-O-acetyloxime

To a suspension of 1.0 g of 11,2'-diacetyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-oleandomycin-30 oxime in 10 ml of benzene is added 0, 18 ml of pyridine followed by 0.24 ml of acetic anhydride. After stirring for 2 hours at room temperature, a further 0.09 ml of pyridine and 0.12 ml of acetic anhydride are added and stirring is continued overnight. The reaction mixture is poured into water and the pH is adjusted to 7.5, then to 9.5 by the addition of solid sodium bicarbonate and 1N sodium hydroxide, respectively. The benzene layer is separated, dried over sodium sulfate 38 6 8 0 5 9 and evaporated to dryness in vacuo to give 890 mg of the desired product.

NMR (δ, CDCl 3): 3.31 (1.5H) s, 3.25 (1.5H) s, 2.25 (6H) s, 2.16 (3H) s, 2.01 (6H) s and 0.55 (4H) m.

In a similar manner, the oximes of Example 35 are converted to their O-acetyl derivatives.

Example 37 11-Acetyl-N-deoxy-N-amino-oleandomycin

To a suspension of 10 g of a 10% palladium-carbon mixture in 100 ml of methanol is added 21.2 g of ammonium acetate and the resulting slurry is treated with 20 g of 11-acetyl-4 "-deoxy-4" -oxo-oleandomycin 100 ml of the same solvent. The suspension is shaken in hydrogen gas at an initial pressure of 50 pounds per square inch (about 2 3.5 kg / cm). After 1.5 hours, the catalyst is filtered off and the filtrate is added to a stirred mixture of 1,200 ml of water and 500 ml of chloroform. The pH is adjusted from 6.4 to 4.5 and the organic layer is separated. The aqueous layer, while still extracted with 500 ml of chloroform, is treated with 500 ml of ethyl acetate and the pH is adjusted to 9.5 with 1N sodium hydroxide. The ethyl acetate-20 layer is separated and the aqueous layer is extracted once more with ethyl acetate. The ethyl acetate extracts are combined, dried over sodium sulfate and concentrated to a yellow foam (18.6 g) which, crystallized from diisopropyl ether, gives 6.85 g of pure product, m.p. 157.5-160 ° C.

NMR (d 1 CDCl 3): 3.41 (3H) s, 2.70 (2H) m, 2.36 (6H) s and 2.10 (3H) s.

The second epimer, which is up to 20-25% in the crude foam, is separated by gradual concentration and filtration of the mother liquors.

Example 38

Following the procedure of Example 37, starting from the appropriate 4'-deoxy-M "-oxo-oleandomycin, the following amines are prepared: 39 68059

RlV N (CH 2) x x i 32

o ^ o / y X

:: K r j 5 Rn - '<x0 ^ \ 0y \ RO ,,, / \

, L .J

10 3 K 2 H 2 O 3 R 3 NMR U, CDCl 3) 0 0 15 CH 3 C-CH 2 C- 3.43 (3H) s, 2.70 C 2 H) m, 2.30 (6H) s and 2.10 (6H) s.

H- H- 5.60 (1H) m, 3.36 (3H) s, 2.83 (2H) m 2 O 0 and 2.30 (6H) s.

H-CH 3 C- 5.80 (1H) m, 3.43 (3H) s, 2.80 (2H) m, 2.30 (6H) s and 2.10 (3H) s.

Example 39 Again following the procedure of Example 37 using the required 4 "-deoxy-4" -oxo-oleandomycin and isopropanol as solvent, the following products are obtained: 2'-propionyl-4 "-deoxy-4" -amino-oleandomycin, 11 -30 acetyl-2'-propionyl-4 "-deoxy-4" -amino-oleandomycin, 11-propionyl-2'-acetyl-4 "-deoxy-4" -amino-oleandomycin and 11,2'-dipropionyl-4 "-deoxy-4" -amino-oleandomycin. Example 40 11-Acetyl-4 "-deoxy-4" -amino-oleandomycin 35 To a suspension of 50 g of 11-acetyl-4 "-deoxy-4" -oxo-oleandomycin and 5 g of ammonium acetate in 500 ml: at -10 ° C methanol, is added dropwise with stirring over an hour.

_________. 1.0 68059 3.7 g of 85% borocyanohydride dissolved in 200 ml of methanol. Stir for two hours in the cold and pour the reaction mixture into 2.5 liters of water and one liter of chloroform. The pH is adjusted to 7.2 to 9.5 by adding 1N sodium hydroxide and the organic layer is separated. The aqueous layer is washed once with chloroform, the organic phases are combined. The chloroform solution of the product is treated with 1.5 liters of water at pH 2.5 and the aqueous layer is separated. The pH of the aqueous phase is adjusted from 2.5 to 7.5 and then to 8.25 and then extracted with ethyl acetate. These extracts are discarded and the pH is finally raised to 9.9. The aqueous layer is extracted (2 x 825 mL) with ethyl acetate and the extracts are combined and dried over sodium sulfate. Removal of the solvent under reduced pressure gives 23.9 g of a foamy product.

NMR (J, CDCl 3): 3.41 (3H) s, 2.70 (2H) m, 2.36 (6H) s and 2.10 (3H) s.

Example 41 4 "-deoxy-9" -amino-oleandomycin A solution of 20 g of 2'-acetyl-4 "-deoxy-4" -oxo-oleandomycin in 125 ml of ethanol is stirred overnight at room temperature and then treated 21, With 2 g of ammonium acate-20 tate. The resulting solution is cooled in an ice bath and treated with 1.26 g of sodium cyanoborohydride. The cooling bath is then removed and the reaction mixture is stirred at room temperature for two hours. The reaction mixture is poured into 600 ml of water and 600 ml of diethyl ether and the pH is adjusted from 8.3 to 7.5. The ether layer is separated and the aqueous layer is extracted with ethyl acetate. The extracts are set aside and the pH of the aqueous layer is adjusted to 8.25. The diethyl ether and ethyl acetate extracts made at this pH are also set aside and the pH is raised to 9.9. The diethyl ether and ethyl acetate extracts made at this pH are combined, washed successively with water (1x) 00 and saturated brine and dried over sodium sulfate. The latter extracts, taken at pH 9.9, are concentrated to a foam which is purified by chromatography on 160 g of silica gel using chloroform as the carrier solvent and the eluent. After collecting 11 fractions, each of 12 ml, the eluent is changed to a mixture of 5% methanol and 95% chloroform. For fraction 370, a mixture of 10% methanol and 90% chloroform is changed to the eluent, and for fraction 440, a mixture of 15% 1.1 68059 methanol and 85% chloroform is used. Fractions at 85-25 ° C are combined and evaporated to dryness in vacuo to give 2.44 g of the desired product.

NMR (δ, CDCl 3): 5.56 (1H) m, 3.36 (3H) s, 2.9 (2H) m and 2.26 δ (6H) s.

Example 42

Following the procedure of Example 40 and starting from the appropriate 4 "-deoxy-4" -oxo-oleandomycin and isopropanol as solvent, the following compounds are prepared: 11,2'-dia-10-cetyl-4M-deoxy "4" -amino-oleandomycin, 2 ' -propionyl-4 "-deoxy-4" -amino-oleandomycin, 2'-acetyl-4 "-deoxy-4" -amino-oleandomycin, 11-acetyl-2'-propionyl-4 "-deoxy-4 '' - amino-oleandomycin, 11-propionyl-2'-acetyl-4 "-deoxy-4" -amino-oleandomycin and 11,2'-dipropionyl-4 "-deoxy-15 4-amino-oleandomycin.

Example 43 4 '' - Deoxy-4 '' -amino-oleandomycin

Stir at room temperature in the presence of nitrogen gas overnight a solution of 300 mg of 2'-acetyl-4 "-20 deoxy-4" -amino-oleandomycin in 25 ml of methanol. The reaction mixture is concentrated in vacuo to give the desired product as a white foam.

NMR δ, CDCl 3): 5.56 (1H) m, 3.36 (3H) s, 2.90 (2H) m and 2.26 (6H) s.

25 Example 44

By repeating the procedure of Example 43 using the required 21-alkanoyl-4 "-deoxy-4" -amino-oleandomycin, the following compounds are obtained: 30 N (CH-) „roc? Λ 35 ^ jp>, / ° ^ 5 Vs' 0CH3 42 _ Λ 68059 Starting material Product - -1- * 0 0 o "·» Il ca3 »> CH3C- ch3c- h- 5 ooo ch3c- ch3ch2c- ch3c- h- 0 0 o ch3ch2c- ca3c- ch3ch2c- u- oo * o 10 "Il m ch3ch2c- ch3ch2c- ch3ch2c- H- 0 u- CH3CH2C- h- h- ^ Example 4 , 8α-dihydro-4 "-deoxy-4" -amino-, oleandomycin

A solution of 2.15 g of 11-acetyl-8,8a-deoxy-8,8a-dihydro-4, '- deoxy-4 "-oxo-oleandomycin and 2.31 g of ammonium acetate in 15 ml: methanol cooled to 20 ° C, treated with 136 mg of sodium cyanoborohydride. It is then stirred for 45 minutes at room temperature and then the reaction mixture is poured into 60 ml of water and 60 ml of diethyl ether and the pH is adjusted to 8.1.

7.5. The ether layer is separated and discarded, and the pH of the aqueous layer is raised to 8.9. Add fresh ether, shake with the water layer and discard the ether layer. Adjust the pH to 8.5 and repeat the recent procedure. Finally, the pH is adjusted to 10 ° C and 60 ml of ethyl acetate are added. The aqueous layer is discarded and the ethyl acetate layer is treated with 60 ml of new water. The pH of the aqueous layer

! adjusted to 6, with 1N hydrochloric acid, and the ethyl acetate layer is discarded. The aqueous layer is extracted successively with pI at 6.5, 7, C, 7.5, 8.0 and 8.5 in ethyl acetate (60 ml) and the organic extracts are discarded. Finally, the pH is raised to 10.0 and the aqueous layer is extracted with ethyl acetate. Combine the extracts collected at pH 8.0, 8.5 and 10.0

II

43 68059 and concentrated in vacuo to give 585 mg of a white foam consisting of a pair of 4 "epimers.

NMR (<f, CDCl 3): 3.38 and 3.35 (3H) 2 singlets, 2.31 and 2.28 (6H) 2 singlets and 2.03 (3H).

Example 46 8,8a-Deoxy-8,8a-dihydro-4 ”-deoxy-4" -amino-oleandomycin

Sodium cyanoborohydride (126 mg) is added to a solution of 1.86 g of 8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin and 2.1 g of am-10 ammonium acetate in 10 ml of methanol at room temperature. temperature. After 1 hour, the reaction mixture is cooled to 0 ° C and stirred for 2.5 hours. The reaction mixture is poured into 60 ml of water and 60 ml of diethyl ether and the pH is adjusted to 7.5. The ether layer is discarded and the pH of the aqueous layer is adjusted first to 8.0 and then to 8.5 by extraction with ether after each adjustment. Finally, the pH of the aqueous layer is adjusted to 10.0 and extraction is performed with ethyl acetate. New water is added to the ethyl acetate extract and its pH is adjusted to 6.0. The ethyl acetate layer is discarded and the pH of the aqueous layer is adjusted successively to 6.5, 8.0, 8.5 and 10.0, and extraction is performed after each pH adjustment with ethyl acetate. The ethyl acetate extracts obtained at pH 7.5, 8.0 and 10.0 are combined and concentrated to a foam which is redissolved in ethyl acetate and extracted with fresh water at pH 5.5. The acidic aqueous layer is adjusted sequentially as above to pH 6.0, 6.5, 7.0, 7.5, 8.0 and 10.0 and extracted after each pH adjustment with diethyl ether. The extracts obtained at pH 7.5, 8.0 and 10.0 are combined and evaporated to dryness in vacuo to give 166 mg of the desired product.

NMR δ, CDCl 3): 5.48 (1H) m, 3.40 (3H) s and 2.30 (6H) s.

30

Example 47

By repeating the procedure of Example 45 using 8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -oxo-oleandomycin as the starting material and isopropanol as the solvent, the following compounds are obtained: 6 8059 n N (CH 2) κι \ ! 3 3 0 ή Τ '! "'W In' '' RO, \ vaO J \ 5 r '·· /

Uvy (ν'1 * »,

ίο I

OCH., N> -

15 H 'S T

CH C- CH C- J n 3 »« CH C1I C- CII C- 20 CH3CH2 ^ - H-

Example 48 11-Acetyl-8,8a-deoxy-8,8a-methylene-U ', - deoxy-U "-amino-oleandomycin 25 300 mg of sodium cyanoborohydride are added to a solution cooled to 20 ° C containing 5.0 l of acetyl-8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-oleandomycin and 5.2 g of ammonium acetate in 30 ml of methanol. The reaction mixture is stirred at room temperature for one hour and then poured into 120 ml of nothing. water and no diethyl ether 120. The pH of the aqueous layer is adjusted successively to 7.5, 8.0, 3.5 and 10.0 and extraction with ethyl acetate is performed after each pH adjustment, and water and pH are added to the final organic extract obtained at pH 10.0. is adjusted to 6. The aqueous layer 35 is treated again as above and the pH is adjusted to 45.08059 by 7.0, 7.5, 8.0, 8.5 and 10.0, extraction with ethyl acetate after each change in pH. ethyl acetate extracts obtained at 8.0, 8.5 and 10.0 and concentrated in vacuo to give 1.5 g of the desired product. s.

NMR (of, CDCl 3): 3.38 (3H) s, 2.30 (6H) s, 2.05 (3H) s and 0.65 (4H) m.

Example 49

The following compounds are prepared using the appropriate 8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-oleandomycin as starting material and isopropanol as solvent and following the procedure of Example 48: R ncch> 2 α Ν'Λ νλ T ¥ «* 0 V LXn„ 2 u 0CH3 R R1 25 0 0 Η 11 CH3-C- CH3C_ 0 0

CH3-C- CH3CH2-C

o o 30 CH3CH2C- ch3-C- 0 o CH3CH2C- CH3CH2C- 0 H- CH.-C- 35 0 H- CH3CH2C- 46 68059

Example 50 11,2 1-Diacetyl-N-deoxy-N-amino-oleandomycin

A suspension of 1 g of Raney nickel washed with isopropanol and 250 mg of 1, 2'-diacetyl-4 "-deoxy-5 4" -oxo-oleandomycin-O-acetyloxime in 25 ml of isopropanol is shaken in hydrogen gas at an initial pressure of 50 2 lbs. / square inch (approximately 3.5 kg / cm) at room temperature overnight. The reaction mixture is filtered and the filtrate is concentrated under reduced pressure to give 201 mg of the desired product.

Boiling to reflux the entire 201 mg in methanol for 1 hour gives 11-acetyl-4 "-deoxy-4" -amino-oleandomycin identical to that prepared in Example 37.

Example 51 Using the starting 4 "-deoxy-4" -oxo-oleandomycin derivative and following the procedure of Example 50 and using the specified catalyst, the following 4 ', - deoxy-4 "-amino-oleandomycins are prepared: 20 1Λ-» ΓΛ' Λ Λ 25 R0 ///;>: XV ", 30 0CH3 47 6 8 0 5 9 R Derivative Catalyst 0 0

"II

CH 2 Cl 2 CH 2 C oxime and 0 ° C

5 CH 2 Cl 2 CH 2 CH 2 C oxime Pd / C

O '

II

CH 2 C-H-oxime ptO

0 2 II

CH-CH 2 C-H- oxime Pt.n 10 J 1 rcu 2

H-oxime l’d / C

0 0

"II

CH3C-CH2C-O-acetyloxy. ^ 15 0 0 II „CH3C-CH3CH2C-O-acetyloxy. M ±

. 0 II

CK3C-O-acetyloxy. Pd / C

0 20 "H-CH 3 C-O-methyloxime pt0 0 0 2

"II

CH 3 C-Cl 2 CH 2 C-O-methyloxime Pd / C

0

II

CE ^ CT ^ C-H-O-methyloxime p ^ / r

Example 52

Following the procedure of Example 50, 8,8a-deoxy-8,8a-dihydro-4 "-deoxy-4" -amino-oleandomycins corresponding to the reduction products of the following 4 "-oxo derivatives are prepared using said catalyst. 68059

R

Υχγ ° ~;

RO I / ((| / '\ Q

5 "n ° Y

l 1 p Y ^ x

o I

OCHo o 10 R X Catalyst 0

H- (OH g C - N-OH Pd / C

0 0 15 CH3CH2C- CH3C “N-OH Ni 0 CH3C- H- N-OH Ni 0 CH3CH2C- H- N-OH Pt02 20 0 H- CH3CH2C- N-OH Ni 0 0 CH3CH2C- ch3c- N-OCH3 Pt02 o

25 H-CH3C- N-OCH3 Pd / C

0

CH3C- H- N-OCH3 Pd / C

O

CH3CH2C- H- N-OCH3 PtO2 30 0 0 0 CHgC- ch3c- N-O-CCH3- PtO2 (continued)

II

68059 49 (continued) R X Catalyst 5 0 0 0 CH3CH2C- CH3C- N-O-CCH3- Ni 0 0

CH3CH2C- H- N-O-CCH3- Pd / C

10 0 0 CH 3 C-H-N-O-CCH 3 - Ni 15 Example 53 Repeating the procedure of Example 50 using 8,8a-deoxy-8,8a-methylene-4 "-deoxy-4" -oxo-oleandomycin as starting material and a catalyst , the following compounds are obtained: N (CH3} 2 25 if

RO V <o \ (] A

fW 'oy 30 T | * 2 ° «3 50 6 8 0 5 9 'r R Output line- Catalyst _ - female 0 0 H ·.

CH 2 C-CH 2 C-oxy imi Ni

• O O

5 CH 2 Cl 2 CH 3 CH 2 O-oxime PtO 2 O 0

11 H

CB 2 Cl 2 C-CH 2 Cl 2 C-oxime PtO 2 O n

H- CH 2 Cl 2 oxime Pd / C

10 0 CH 2 Cl 2 -H-oxime PtO 2

o O

Il 1 «

CH 2 CH 2 -CCH 2 O-methyloxime Pd / C

O

15 ..

CH-jC-M-O-methyloxime lil

O O

CH 2 -CCH 2 C-O-acetyloxime Ni 0

II

CH3C-H-O-acetyloxime Ni

O O

H 11

CH 2 C-CH 2 C-O-acetyloxime Pd / C

Example 54 25. ...

4 "-deoxy-4" -ethyl-oleandomycin

To a mixture of 4.59 g of 4 "-deoxy-4" -oxo-olendomycin, 6.6 ml of a 5-molar ethanolic solution of ethylamine and 1.89 ml of acetic acid in 25 ml of methanol is added 365 mg of sodium cyano-30-borohydride. In doses of 50-60 mg. Stir for 1 hour at room temperature and then pour the reaction mixture into 110 ml of water and 120 ml of ethyl acetate.

The pH of the aqueous layer is adjusted successively to 7.5, 8.0, 8.5 and 10.0 and extraction is performed with ethyl acetate after each pH adjustment. The last organic extract obtained in pI 10.0 is treated with water and adjusted to pH 6. The aqueous layer is treated again as above and the pH is adjusted to 7.0, 7.5, 8.0, 8.5 and 10.0 by extraction with ethyl acetate after each pH change. The ethyl acetate extracts obtained at pH 8.0, 8.5 and 10.0 are combined and concentrated in vacuo to a foam. The product is further purified by chromatography on 75 g of silica gel using acetone as eluent. Fractions 62-104, each containing 4 ml, are combined and concentrated under reduced pressure to give 910 mg of the desired product.

Example 55 11-Acetyl-4,1-deoxy-4 "-ethylamino-oleandomycin

In a manner similar to Example 54, 366 mg of sodium cyanoborohydride are added portionwise to a solution of 5.82 g of β-11-acetyl-4 "-deoxyoxooleandomycin and 16 ml of a 5 molar ethanol solution of ethylamine in 27.4 ml of hydrogen chloride 2. After stirring at room temperature for 1.5 hours, the reaction mixture is poured into 120 ml of water and 120 ml of ethyl acetate and treated as in Example 54 to give 1.2 g of the desired product.

Example 56 11-Acetyl-4H-deoxy-4H-n-hexylamino-oleandomycin

Following the procedure of Example 54, starting from 4.8 g of 11-acetyl-4 "-deoxy-4" -oxo-oleandomycin, 6.7 g of n-hexylamine, 3.78 ml of acetic acid, 302 mg of sodium cyanoborohydride and 25 g of starting material. ml of ethanol, 80 g of silica gel are purified by chromatography using 1.3 g of the desired product as eluent.

Example 57 Using the appropriate 4 "-deoxy-4" -oxo-oleandomycin and amine and isopropanol as solvent as starting materials and following the procedure of Example 54, the following compounds are prepared: 35 52 68059 RK NCCH3) 2 ° Λ ° Λ

RO «/„ Y

^ j5V ° Y

^ T | o I 0 OCH3 10 - Rj R3 0 15 CH32 "H- CH3- 0 CH3C- H- n-C3H? - 0 CH, C- H- iC" H "- 20 d - J / 0 CH3C- H- B" C5H11 "0 0

tl M

CH3C- CH3C_ C2H5 "25 0 0 CH3C- ch3C- i-C3H? - 0 0 CH3C- ch3C- t-C4Hg- 30 0 0 ch3c- ch3ch2c- ch3- 0 0 CH3C- CH3CH2C- n-CgH13- (continued) 35 53 68059 (continued) R 2 R 3

Ö O

Il II

5 CH3C- CH3CH2C- itaC5Hn "

O

II

CH3CH2C- h- n “C3ri7 ~

O

tl CH-CH-C- H- n-C, H0- 3 2 - 4 9

10 O

11 CH3CH2C- h- n-C6li13-

0 O

Il II

CH-CH „C- CH-C- CH-- J t. Λ 3 O o 15 CH3CH2C- CH3C-

O O

Il tl CH3CH2C- CH3C- n-C5ni; L-

0 O

20 CH3CV- CH3 ^ - - "C5H11" O o "1.

CH3CH2C- CH2CH2C- C2H5- 0 o

Il II

CH.CK-C- CH_CH., C- n-Cr H1 3 2 3 2 - o 13 25 0 0

H II

CH3CH2C- CH3CH2C- i “C4H9 ~ H- H- CH3 ~ 30 H- H- C2H5- H- K- t-C4K9- H- H- -“ S ”li 'H- H- n-C, H.

- o 1j 35

Si, 68059

Example 58 11-Acetyl- * 4 "-deoxy-4H-dimethylamino-oleandomycin 2 g of 11-acetyl-4" -deoxy-4 "-amino-oleandomycin, 5 g of 10% palladium-on-carbon and 2.06 ml of the formalin solution are mixed with 4 to 0 ml of methanol and shaken in hydrogen gas overnight at an initial pressure of 50 pounds per square inch (about 3.5 kg / cm 2), the spent catalyst is filtered off and the filtrate is evaporated to dryness under reduced pressure. The residual product (1.97 g) is purified by chromatography on 40 g of silica gel using chloroform as eluent, after 25 fractions comprising 650 drops each, a 3% solution of methanol in chloroform is exchanged as eluent, and fractions 36-150 are combined and concentrated in vacuo. to give 704 mg of the desired product as a white foam.

NMR (δ, CDCl 3): 3.33 (3H) s, 2.63 (2H) m, 2.30 (12H) s and 210 (3H) s.

Example 59 Following the procedure of Example 58, except that isopropanol is used as the solvent and 4 "-deoxy-4" -amino-oleandomycin is used as the starting material, the following compounds are prepared: 25. R.

"Fixx

RO *, / V'Cs'- '”"

0 I x CH

0CH3 un3 35 il 55 6 8 0 5 9 -! I ° o

"M

CH3C- CH3C- 0 o "u CH3C" CH3CH2C-

5 O

'Il H- CH3C-

0 O

ch3ch2c- ch3c- 0 10 CH3CH2C- k- 0 0 CH3CH2C- CH3CH2C- H- H- 15

Example 60 11-Acetyl-4 "-deoxy-β-amino-oleandomycin dihydrochloride To 7.28 g of 11-acetyl-4'-deoxy-4" -amino-oleandomycin in 50 ml of dry ethyl acetate is added 20 ml. a 1N ethyl acetate solution of hydrogen chloride and the solution thus obtained is evaporated to dryness under reduced pressure. The residue is crystallized by trituration from ether and filtered to give the desired salt.

In a similar manner, the amine-25 compounds of the present invention are converted to their diacid addition salts.

Example 61 11,2,11-Diacetyl-N, N-deoxy-N-amino-oleandomycin hydrochloride

The procedure of Example 60 is followed with the exception that 10 ml of 1-n ethyl acetate solution of hydrogen chloride are added. The solution is evaporated to dryness in vacuo and the remaining monohydrochloride salt is crystallized by trituration from ether and filtered.

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

55 6 80 5 9

Example 62 Asparaginate of 11-acetyl-4'-deoxy-u "-amino-oleandomycin To 960 mg of 11-acetyl-4" -deoxy-4 "-amino-oleandomycin in 6 ml of 40 ° C acetone is added 18 ml of water. and then 175 mg of as-5 paraginic acid. The mixture is refluxed until a cloudy solution is obtained. The mixture is filtered hot and the clear filtrate is concentrated to remove acetone. The remaining solution is lyophilized to leave the desired product.

Preparation A

10 2-Acetyl-8,8a-deoxy-8,8a-dihydro-oleandomycin 1a. 21-Acetyl-8,8a-deoxy-oleandomycin

Zinc dust (10 g) and mercury 2) chloride (1 g) are placed in a round 250 ml three-necked flask. After the solids are mixed well, 1N HCl (25 mL) is added and the mixture is stirred vigorously for 15 minutes. The supernatant is removed from the surface of the water and fresh HCl (25 mL) is added and carbon dioxide is introduced into the flask. A filtered solution of chromium trichloride (50 g in 65 mL of 1 N HCl) is added rapidly to the zinc amalgam. The mixture is stirred in carbon dioxide gas for one hour, at which time the resulting light blue color indicates the presence of chromium 2) chloride (CrCl 2). Stirring is stopped after one hour and the zinc amalgam is allowed to settle to the bottom of the flask.

2'-Acetyl-oleandomycin (29.2 g) dissolved in acetone (200 ml) and water (100 ml) is placed in a round dropping funnel attached to a 600 ml three-necked flask equipped with an overhead mechanical stirrer. With constant stirring in the presence of carbon dioxide, 2'-acetyl-oleandomycin solution and the previously prepared chromium (2) chloride solution are added to this flask. The solution is added at the same time so that both run out at the same time. The Li-30 addition takes about 12 minutes. Stir for 35 minutes at room temperature and then add water (100 ml) and ethyl acetate (100 ml) to the reaction mixture and continue stirring for 15 minutes. The ethyl acetate layer is separated and washed with water (80 ml). The ethyl acetate is separated and the extracts are combined and washed with fresh ethyl acetate (100 mL). The ethyl acetate layer is separated and washed with water (100 mL). The organic phase is separated and the aqueous phases are combined and treated with sodium chloride (75 g). The still separating ethyl acetate is discarded and combined with other ethyl acetate extracts. Water is added to the combined ethyl acetate extracts and the pH is adjusted to 8.5 with sodium bicarbonate. The organic layer was separated, washed with water, saturated sodium chloride and dried over anhydrous sodium sulfate. Filtration and evaporation of the solvents under reduced pressure gives a white solid which is crystallized from ethyl acetate / heptane to give the desired product, 8.4 g, m.p. 183.5-185 ° C.

10 Analysis for C37H63013N:

Calculated: C 62.2 H 9.0 N 2.0 Found: C 62.0 H 8.9 N 2.0.

NMR (δ, CDCl 3): 5.63 (1H) s, 5.58 (1H) s, 3.43 (3H) s, 2.36 (6H) s and 2.08 (3H) s.

15 1b. 2 * -acetyl-8,8a-deoxy-8,8a-dihydro-oleandomycin

The aluminum foil (4.0 g) cut into 1/4 inch pieces is stirred for 30-45 seconds in 290 ml of an aqueous solution of mercury (2) chloride. The solution is decanted and the amalgamated aluminum is washed successively with water (2x), isopropanol (1x) and tetrahydrofuran (1x). 45 ml of tetrahydrofuran, 45 ml of isopropanol and 10 ml of water are poured on top of the pieces and then cooled to 0 [deg.] C. in an ice bath. 2.0 g of 8,8a-deoxy-2'-acetyl-oleandomycin dissolved in tetrahydrofuran, isopropanol and water are added dropwise to the amalgamated aluminum so that the temperature is maintained at 0 ° C. After the addition is complete, the ice bath is removed and the mixture is stirred at room temperature overnight. The solids are filtered off and the filtrate is evaporated to dryness in vacuo. The residue is treated with ethyl acetate-water and the pH is adjusted to 9.0 with saturated sodium carbonate solution. The organic phase is separated, washed with water and brine, and dried over sodium sulfate. Removal of the solvent gives 2.27 g of the desired product.

58

Preparation B 68059 2a. 11,2'-Diacetyl-8,8a-deoxy-8,8a-methylene-oleandomycin

To a flame-dried 200 ml three-necked flask equipped with a dropping funnel, a magnetic stirrer and a nitrogen gas charge is placed 16.4 g of trimethylsulfoxonium iodide and 3.4 g of a 50% oil dispersion of sodium hydride. The solids are mixed well and 43.2 ml of dimethyl sulfoxide are added via addition funnel. One hour after the evolution of hydrogen has ceased, the suspension is cooled to 5-10 ° C and 22.6 g of 11,2 * -diacetyl-8,8a-deoxyoleandomycin dissolved in 32 ml of tetrahydrofuran are added over 10 minutes and To 16 ml of dimethyl sulfoxide. The suspension is stirred at room temperature for 90 minutes and then poured into 300 ml of water and extracted with two 300 ml portions of ethyl acetate. The organic extracts are washed with water and saturated sodium hydroxide solution, dried over anhydrous sodium sulfate, filtered and evaporated to dryness under reduced pressure. The residue is crystallized from ether to give 8.9 g of 11,2'-diacetyl-8,8a-deoxy-8,8a-methylene oleandomycin.

20

II

Claims (7)

A process for the preparation of therapeutically useful 4 "amino-oleandomycin derivatives of formula IV, V or VI 5 and pharmaceutically acceptable acid addition salts thereof, R. N (CHJ" ro \ | 32 R "ab y \ 0 ° r \ vsN * A ° Ύ 0N> IdT ^ Γον ° γ T NHR. [0 \ || NH „15 ° ®3 0 T 2 IV V« YR, N (CH0) „° ΛΧΛ 2. R0 <'h, ^ 71 NH0 I I AND Wherein R vilka and R ^ are hydrogen or alkanoyl having two or three carbon atoms; R 3 is hydrogen or alkyl of 1-6 carbon atoms, characterized in that i) a compound of formula I, II or III is reduced to N (CH 3> 2 Vk. > 2 R0 "'f> ° ^ cX 5. V Y Ύ V cj, T 0CHq 10. II 3 N (CH3) 2 w RO Uv J J 'dl ,, / V) - ^ N X and q are 111 where R and each are hydrogen or alkanoyls having two or three carbon atoms and X is NR 2, N-OH, N-OCH 2 or N-2. OC-CH 2, such that 0 a) where X is N-OH, N-OCH 2 or N-OC-CH 2, said reduction is carried out by catalytic hydrogenation and 0 b) when X is NR, this imino group is formed from corresponding ketones (X = 0) of formula I, II or III by condensing said ketone with an amine or ammonium salt of a lower alkanoic acid or with an ammonium or amine salt of an inorganic acid, and the reduction, which is a hydride reduction, is carried out in situ and if desired, ii) where R or R 2 is hydrogen, the respective ones are converted to alkanoyl and / or where R or R 4 is alkanoyl, the respective ones are converted to hydrogen; and / or ii) pharmaceutically acceptable acid addition salts are formed.