NO145384B - ANALOGY PROCEDURE FOR THE PREPARATION OF THERAPEUTIC ACTIVE OLEANDOMYCIN DERIVATIVES - Google Patents

Description

This invention relates to an analogous process for the preparation of new antibacterial compounds and in particular to a series of 4"-deoxy-4"-sulfonylamino-oleandomycins and their pharmaceutically acceptable acid addition salts.

Oleandomycin, its preparation in fermentation broth and its use as an antibacterial agent was first described in US Patent 2,757,123. It is known that the naturally occurring compound has the following structure:

The conventionally accepted numbering system and the stereochemical notation for oleandomycin and similar compounds are shown in a number of positions.

US Patents 3,884,902 and 3,983,103 relate respectively to 4"-erythromycin sulfonate esters and N-sulfonylerythromycin amines which have biological profiles different from the compounds produced according to the present invention.

Several synthetic modifications of oleandomycin are known, especially those where from one to three of the free hydroxyl groups in the 2'-,

The 4" and 11-positions are esterified as acetyl esters. In addition, US patent 3,022,219 describes similar modifications where the acetyl groups in the above-mentioned esters are replaced with another, preferably unbranched, lower alkanoyl group with 3 to 6 carbon atoms.

The semi-synthetic oleandomycin antibacterial agents

which is produced according to the invention, is covered by the formula:

and

the pharmaceutically acceptable acid addition salts thereof, wherein R is alkyl having from 1 to 3 carbon atoms; pyridyl; 1,1,1-trifluoroethyl; phenyl; monosubstituted phenyl wherein the substituent is selected from fluorine, chlorine, bromine, iodo, hydroxy, methoxy, cyano, carboxamido, nitro, amino, carbomethoxy, carbobenzyloxy, carboxy, trifluoromethyl, alkyl having from 1 to 4 carbon atoms and acetamido; disubstituted phenyl wherein the substituents are each selected from chloro, nitro, amino, methoxy and methyl; trichlorophenyl; hydroxydichlorophenyl; benzyl'; naphthyl; thienyl; chlorothienyl; 2-acetamido-5-thiazolyl; 2-acetamido-4-methyl-5-thiazolyl; 2-benzimidazolyl; dimethyl-2-pyrimidinyl; pyrryl; furyl; monosubstituted thienyl, pyrryl and furyl wherein the substituent is selected from carbomethoxy and alkyl of 1 to 2 carbon atoms; or 1-methyl-5-carbomethoxy-3-pyrryl; and

is hydrogen or alkanoyl of 2 to 3 carbon atoms;

provided that when. R-1 is hydrogen, R is phenyl; thienyl; mono-substituted phenyl wherein the substituent is selected from chloro, fluoro, methyl, methoxy and trifluoromethyl; or alkyl substituted thienyl where the alkyl group has from 1 to 2 carbon atoms.

The amine starting materials used in the production of the new compounds comprise, due to the synthesis method used for their production, 4"-epimeric amines. The new sulfonamido compounds produced from said amines thus also comprise an epimeric mixture. Experimentally, it has been found that both epimeric sulfonamides are present in the final product in varying proportions depending on the choice of synthesis method used to prepare the amine intermediate. If the isolated product consists predominantly of one of the epimers, said epimer can be purified by repeated recrystallization from a suitable solvent of constant melting point. The other epimer, which is present in smaller amounts in the initially isolated solid material, is the predominant product in the mother liquor. It can be recovered from this by methods known per se, such as f .eg by evaporation of the mother liquor and repeated recrystallization of the residue into a product with a constant melting point or d chromatography.

Although the mixture of epimers can be separated by methods known per se, for practical reasons it is appropriate to use the mixture as it is isolated from the reaction mixture. However, it is often convenient to purify the mixture of epimers by at least one recrystallization from a suitable solvent, by subjecting it to column chromatography, solvent partitioning or by trituration in a suitable solvent. This purification, which does not necessarily separate the epimers, leads to the removal of such foreign materials as starting materials and unwanted by-products.

: The absolute stereochemical determination of the epimers has not been completed. However, both epimers of a given compound exhibit the same type of activity, e.g. as antibacterial

Preferred compounds of formula 1 if those where R is substituted phenyl, thienyl and substituted thienyl where the substituent is alkyl with from 1 to 2 carbon atoms or carbomethoxy.

Preferred among these compounds because of their antibacterial properties are 11-acetyl-4"-deoxy-4"-(2-thienylsulfonylamino)oleandomycin, 11-acetyl-4"-deoxy-4"-(3-thienylsulfonylamino)oleandomycin, 11 -acetyl-4"-deoxy-4"-(3-methyl-2-thienylsulfonylamino)oleandomycin, 4"-deoxy-4"-(p-chlorophenylsulfonylamino)-oleandomycin, 4"-deoxy-4"-(2-thienylsulfonylamino )-oleandomycin, 4"-deoxy-4"-(3-thienylsulfonylamino)oleandomycin, and 4"-deoxy-4"-(3-methyl-2-thienylsulfonylamino)oleandomycin.

The method according to the invention for the production of 4"-deoxy-4"-sulfonylamino-oleandomycin-derived antibacterial agents of formula _1 by starting with 4"-deoxy-4"-amino-oleandomycin or an 11-alkanoyl derivative thereof, can be illustrated by means of

the following form:

where R and R 1 are as previously indicated, and W is a halogen atom.

The above reactions are carried out between a 4"-deoxy-4"-amino-oleandomycin and an appropriate sulfonyl halide in the presence of an acid scavenger in a reaction-initiated solvent.

In practice, one mole of 4"-amino-oleandomycin is contacted with one mole of the sulfonyl halide plus as much as a 2-3% excess

of said halide. The acid scavenger, which may be inorganic or organic in nature, is used in an amount of one mole plus as much as 4-6% excess.

The acid scavenger may be alkali metal or alkaline earth metal hydroxides, hydrides or carbonates as well as a tertiary organic amine. In addition, one can also use secondary amines, such as diisopropylamine, which are sufficiently hindered so that they do not react with the sulfonyl halide starting material.

The preferred class of acid scavengers are tertiary amines. Particularly preferred within this class is triethylamine.

The reaction-initiated solvent used in the above-mentioned method should be one that substantially solubilizes the reaction components and does not. reacts to any significant extent with either the reaction components or the products formed. Particularly preferred are polar solvents which are miscible or immiscible with water. Methylene chloride and acetone-water are particularly preferred.

Since ppp heating of amino-oleandomycins -leads to

a certain cleavage, it is preferred that the method for preparing the compounds with formal is carried out at 0-25°C. It is particularly preferred that the turnover is carried out at ambient or room temperature.

The reaction time is not critical and depends on the reaction temperature, concentration and inherent reactivity of the starting materials. When the reaction is carried out at room temperature using the concentrations indicated below, the reaction is essentially complete after 2 to 48 hours.

After completion of the reaction, the reaction mixture can be worked up by one of two methods, both of which are known to those skilled in the art. The first work-up method involves adding the reaction mixture to water, followed by separation of the non-water-miscible. solvent containing the desired product, and removal of this then yields the crude product. When a water-miscible solvent is used as the reaction-initiated solvent, the product is extracted from the aqueous reaction mixture using a water-immiscible solvent such as methylene chloride.

The other processing method includes concentration

of the reaction mixture to dryness followed by extraction of the product from the salt resulting from the acid-trapping base and hydrogen halide by-product, using acetone. The acetone extract can be concentrated to give a crude product.

The crude product or an acetone solution thereof is purified by chromatography on silica gel, which is a well-known method, or recrystallization.

The '.'r-4"-amino starting compounds used in the synthesis of the antibacterial agents are prepared by oxidation of the natural oleandomycin, followed by a reductive amination of the resulting ketone as described below.

By utilizing the chemotherapeutic effect of the compounds produced according to the invention, which form salts,

it is of course preferred to use pharmaceutically acceptable salts.

Although water insolubility, high toxicity or lack of

• crystalline nature can make some particular types of salt unsuitable or less desirable for use as such in a pharmaceutical preparation, water-insoluble or toxic salts can be converted into the corresponding pharmaceutically acceptable bases by cleavage of the salt with a suitable base, or alternatively they can be converted into a any desired pharmaceutically acceptable acid addition salt. Examples of acids which provide pharmaceutically acceptable anions are hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid, sulfuric acid, phosphoric acid. -acetic acid, lactic acid, citric acid, tartaric acid, succinic acid, maleic acid, gluconic acid, aspartic acid, glutamic acid, pyroglutamic acid and laurylsulphuric acid*. The new 4"^deoxy-4"-amino-oleandomycin derivatives prepared according to the invention exhibit in vitro activity against various Gram-positive microorganisms such as Staphylococcus aureus and Streptococcus pyogenes, and against certain Gram-negative microorganisms such as those that have a spherical or ellipsoid shape (cocci). Their activity is easily demonstrated by in vitro tests against various micro-organisms in a brain-heart infusion medium by the usual two-fold serial dilution technique. Their in vitro ■'"activity makes them useful for local application in the form of ointments, . .creams, etc., for sterilization purposes, e.g.*., hospital equipment, and "as industrial antimicrobial agents, e.g. for water treatment, slime control, paint and wood preservation.

For in vitro use, e.g. for local administration, it will often be convenient to mix the selected product with a pharmaceutically acceptable carrier such as a vegetable oil or a mineral oil or an emollient cream. Likewise, the products can be dissolved or dispersed in liquid carriers or solvents such as water, alcohol, glycols or mixtures thereof, or other pharmaceutically acceptable inert media, i.e. media that have no harmful effect on the active ingredient. For such purposes, it will usually be acceptable to use concentrations of the active ingredients of from approx. 0.01 to approx. 10% by weight, based on the total preparation.

Moreover, many of the compounds produced according to the invention are active against Gram-positive microorganisms when administered orally and/or parenterally to animals, including humans. Their

in vivo activity is more limited with respect to organisms affected, and is determined by the usual method which involves infecting mice of approximately equal weight with the test organism and then treating them orally or subcutaneously with the test compound. In practice, the mice are given, e.g. 10, an intraperitoneal inoculation of suitable diluted cultures containing approximately 1 to 10 times the LD^q0 (the lowest concentration of organisms necessary to produce 100% mortality). Control experiments are carried out simultaneously with mice receiving inoculum in lower dilutions as a check for possible variations in the strength of the test organism. The test compound is administered 0.5 hour after inoculation and repeated 4, 24 and 48 hours later. Surviving mice are kept for 4 days after the last treatment, and the number of survivors is recorded.

When used in vivo, these new compounds can be administered orally or parenterally, e.g. by subcutaneous or intramuscular injection, in a dose from approx. 1 to approx. 200 mg/kg body weight per day. The preferred dose range is from approx. 2 to approx. 100 mg/kg body weight per day, and the particularly preferred area is from approx. 2 to approx. 50 mg/kg body weight per day. Carriers suitable for parenteral injection can either be aqueous, such as water, isotonic saline, isotonic dextrose or Ringer's solution, or non-aqueous, such as fatty oils of vegetable origin (cottonseed, peanut oil, corn, sesame). , dimethyl sulphoxide and other non-aqueous carriers which will not affect the therapeutic effect of the preparation and are non-toxic in the amount used (glycerol, propylene glycol, sorbitol). Preparations which are suitable for the preparation of solutions on the spot before administration can also be appropriately prepared. Such preparations may contain liquid diluents, e.g. propylene glycol, diethyl carbonate, glycerol, sorbitol, etc.; buffer agents, hyaluronidase, local anesthetics and inorganic salts to achieve desired pharmacological properties. These compounds can also be mixed with various pharmaceutically acceptable inert carriers including solid diluents, aqueous carriers, and non-toxic organic solvents in the form of capsules, tablets, pills, lozenges, dry mixes, suspensions, solutions, elixirs and parenteral solutions or suspensions. In general, the compounds are used in different dosage forms in concentrations varying from approx. 0.5 to approx. 90% by weight of the total preparation.

The following examples shall serve to further illustrate the invention.

Example 1

ll- acetyl- 4"- deoxy- 4"-( 2- thienylsulfonylamino) oleandomycin

To 30 ml of dry methylene chloride are added 2.9 g (4.0 mmol) of 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 740 mg (4.1 mmol) of 2-thienylsulfonyl chloride and 0.58 ml (4 .2 mmol) of triethylamine, and the resulting mixture is stirred at room temperature for 18 hours. The reaction mixture is poured into 50 ml of water and then washed with a saturated salt solution and dried over sodium sulfate. The solvent is removed under reduced pressure, and the remaining foam is purified by chromatography over a silica gel column using acetone as solvent and eluent. The fractions containing the product are collected and concentrated in vacuo to dryness, 1.3 g..

NMR (6, CDCl 3 ): 2.03 (3H) s, 2.30 (6H) s, 2.63 (2H) d, 3.16 (3H) s and 6.8-7.8 (3H) m .

Example 2

By starting with 11-acetyl-4"-deoxy-4"-amino-oleandomycin the appropriate sulfonyl chloride and using the procedure of Example 1, the following compounds are prepared:

Example 3

11-acetyl-4"-deoxy-4"-(p-chlorophenylsulfonylamino)oleandomycin

865 mg ( 4.1 mmol) of p-chlorophenylsulfonyl chloride, and the resulting reaction mixture is stirred at room temperature overnight. The reaction mixture is concentrated to dryness in vacuo, and the residue is treated with 10 mL of acetone. The suspension is filtered, and the filtrate is chromatographed on 160 g of silica gel using acetone as eluent. Fractions 51 to 6 3 each comprising 10 ml are collected and concentrated under reduced pressure to give 857 mg of pure product.Fractions 42-52 and 64-92 give 1.21 g of less pure product.

NMR (6, CDCl 3 ):;2.13 (3H) s, 2.36 (6H) s, 2.73 (2H) d, 3.13 (3H) s and 7.3-8.2 (4H) Wq. 20 g of 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 7.24 g of p-chlorophenylsulfonyl chloride and 5.36 g of triethylamine in a solvent system comprising 350 ml of acetone and 350 ml of water gives correspondingly 17.1 g of the desired product which crystallizes from the reaction mixture, m.p. 202-203.5°C. The analytical sample is recrystallized from ethanol/water.

Example 4

Reversing the procedure according to Example 3 and starting with the appropriate sulfonyl chloride and 11-acetyl-4"-deoxy-4"-amino-oleand@myjGin, the following compounds are prepared:

Example 5 V.

ll- acetyl- 4 "■^ Heoxy- 4 "-( o- tolylsulfonylamino) oleandomycin

A solution of 2.9 g (4.0 mmol) of 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 780 mg (4.1 mmol) of o-tolylsulfonyl chloride and

0.58 ml (4.2 mmol) of triethylamine in 30 ml of methylene chloride is stirred at room temperature for 48 hours. The reaction mixture is quenched in 50 ml of water, and the separated organic layer is washed with a saturated salt solution and dried over sodium sulphate. The solvent is removed. in vacuum, and the remaining yellow foam is chromatographed on 200 g of silica gel in a column with a diameter of 3 cm. The product is eluted

from the column with acetone and collected in 10 ml fractions.: The : v fractions containing the pure product, as detected by thin layer chromatography, are collected and concentrated to dryness under reduced pressure to give 1.3 g.

NMR (6, CDCl 3 ): 2.06 (3H) s, 2.33 (6H) s, 2.46 (2H) d, 2.73 (3H) s

and 7.1-8.2 (4H) m.

Example 6

The procedure according to Example 5 is repeated, starting with the appropriate sulfonyl chloride and 11-acetyl-4"-deoxy-4"-amino-oleandomycin, to prepare the following compounds:

Example 7

ll- acetyl- 4"- deoxy- 4"- phenylsulfonylamino- oleandomycin

To a solution of 2.91 g (4.0 mmol) of 11-acetyl-4"-.deoxy-4"-amino-oleandomycin and 424 mg (4.2 mmol) of triethylamine in 30 ml of methylene chloride which is kept cooled in a ice bath, add 722 mg (4.1 mmol) of benzylsulfonyl chloride. After 10 minutes, the bath is removed, and the reaction mixture is stirred at room temperature overnight. The reaction mixture is quenched with 50 ml of water, then the organic layer is washed with a saturated salt solution and dried over sodium sulphate. Removal of the solvent gives the crude product which is further purified by chromatography over 160 g of silica gel using acetone as eluent. Fractions (10 ml each) 61-93 containing the pure product as detected by thin layer chromatography are pooled and concentrated and dried under reduced pressure to give 1.5 g of the desired product.

NMR (6, CDCl'3) : 2.06 (3H) s, 2.30 (6H) s, 2.63 (2H) d, 3.06 (3H) s, and 7.3-8.2 ( 5H) m.

o

In the method according to example 7, the following compounds are also produced using the appropriate starting materials:

11-acetyl-4"-deoxy-4"-(2-naphthylsulfonylamino)oleandomycin

NMR (6, CDCl 3 ): 2.03 (3H) s, 2.26 (6H) s, 2.65 (2H) d, 2.96 (3H)

and 7.4-8.6 (7H) m; and 11- acetyl- 4"- deoxy- 4"- benzylsulfonylamino- oleandomycin NMR (6, CDCl 3 ): 2.00 (3H) s, 2.30 (6H) s, 2.63 (2H) d, 3.46 (3H) s, 4.33 (2H) s, and 7.36 (5H) s.

Example 8

ll- acetyl- 4"- deoxy- 4"-( p- benzyloxycarbonylphenylsulfonylamino)-oleandomycin

A solution of 2.55 g (3.5 mmol) of 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 1.12 g (3.6 mmol) of p-benzyloxycarbonyl-phenylsulfonyl chloride and 379 mg (3, 75 mmol) of triethylamine in 25 ml of methylene chloride is stirred at room temperature overnight. The solvent is removed in vacuo, and the residue is triturated in 10 ml of acetone.

The solids are filtered, and the filtrate is chromatographed on

280 g of silica gel using acetone as eluent and with fractional quantities of 10 ml. Fractions 90-203, as in thin-

layer chromatography is shown to contain most of the pure product, is collected and concentrated under reduced pressure to give 1.25 g of the desired product.

NMR (6, CDCl 3 ): 2.04 (3H) s, 2.30 (6H) s, 2.66 (2H) d, 3.01 (3H) s, 5.48 (2H) s, 7.50 (5H) s and 8.03-8.53 (4H) m.

Example 9

By starting with the appropriate sulfonyl chloride and 11-acetyl-4"-deoxy-4"-amino-oleandomycin and using the method according to example <8> r, the following compounds are prepared:

Example 10

ll- acetyl- 4"- deoxy- 4"-( p- carboxyphenylsulfonylamino) oleandomycin

A suspension of 400 mg of 10% palladium-on-charcoal in 40 ml of ethyl acetate containing 800 mg of 11-acetyl-4"-deoxy-4"-(p-benzyloxycarbonylphenylsulfonylamino)oleandomycin is shaken in a hydrogen atmosphere at an initial pressure of 3.5 kg /cm 2 at room temperature for 2 hours. A further amount of 250 mg of catalyst is added and the reaction is continued for 2 hours.

The spent catalyst is filtered off, and the solvent is removed

in vacuo to give 450 mg of the desired product.

NMR (6, CDCl 3 ): 2.06 (3H) s, 2.86 (6H) s, 2.68 (2H) d, 3.30 (3H) s, and 7.5-8.4 (4H ) m.

Example 11

11-acetyl-4"-deoxy-4"-(o-nitrophenylsulfonylamino) oleandomycin

5 g (6.8 mmol) 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 1.5 g (7.0 mmol) o-nitrobenzenesulfonyl chloride and 0.98 ml triethylamine are mixed in 50 ml methylene chloride and stirred at room temperature for 48 hours. The reaction mixture is quenched with an equal volume of water, and the organic phase is washed with a saturated salt solution and dried over sodium sulfate. Removal of the solvent under reduced pressure gives the crude product as a foam. The product is purified by chromatography on 140 g of silica gel in a column with a diameter of 3 cm, using acetone as eluent. Fractions 20-30, each comprising 50 ml, are collected, mixed and concentrated to dryness to give 3.4 g of the desired compound.

NMR (6, CDCl 3 ): 2.10 (3H) s, 2.33 (6H) s, 4.36 (2H) d, 2.90 (3H) s and 7.4-8.4 (4H) m .

Using the appropriate starting materials and

the above-mentioned method is prepared correspondingly to the following compounds:

11- acetyl- 4"- deoxy- 4"-( m- nitrophenylsulfonylamino) oleandomycin NMR (6, CDCl 3 ): 2.06 (3H) s, 2.30 (6H) s, 2.66 (2H) d, 3 .06 (3H) s, and 7.4-9.0 (4H) m; and 11-acetyl-4"-deoxy-4"-(p-nitrophenylsulfonylamino)oleandomycin NMR (6, CDCl 3 ): 2.10 (3H) s, 2.35 (6H) s, 2.68 (2H) d, 3.06 (3H) s, and 8.0-8.6 (4H) m.

Example 12

ll- acetyl- 4"- deoxy- 4" (p- hydroxyphenylsulfonylamino) oleandomycin

A solution of 2.55 g (3.5 mmol) of 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 701 mg (3.65 mmol) of p-hydroxyphenyl-sulfonyl chloride and 51.8 yl of triethylamine in 25 ml of methylene chloride is stirred at room temperature for 48 hours. The solvent is removed in vacuo, and the residue is treated with 10 ml of acetone. The insoluble components are filtered off, and the filtrate is chromatographed over T.. 200 g of silica gel using acetone as eluent. Fractions 116-175, which by thin-layer chromatography are shown to contain the pure product, are collected and concentrated to dryness under reduced pressure to give 550 mg of the desired product.

NMR (6, CDCl 3 ): 2.0 (3H) s, 2.33 (6H) s, 2.68 (2H) d, 3.06 (3H) s

and 6.6-8.0 (4H) m.

Example 13

ll- acetyl- 4"- deoxy- 4"-( m-carboxamidophenysulfonylamino) oleandomycin

898 mg (4.1 mmol) m -carboxamidophenylsulfonyl chloride, and the resulting reaction mixture is stirred for 48 hours. The solvent is removed in vacuo, and the residue is treated with 25 ml of acetone. The triethylamine hydrochloride is filtered off, and the filtrate is chromatographed on 160 g of silica gel. Fractions, each containing 50 ml, are collected and examined by thin-layer chromatography to determine the purity of the product. Fractions 66-93 are combined and concentrated under reduced pressure to give 800 mg of the desired product.

NMR (6, CDCl 3 ): 2.06 (3H) s, 2.33 (6H) s, 2.70 (2H) s, 3.10 (3H) s and 7.4-9.0 (4H) m .

Example l 14

ll- acetyl- 4"- deoxy- 4"-( p- acetamidophenylsulfonylamino) oleandomycin

A solution of 2.91 g (4.0 mmol) of 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 955 mg (4.1 mmol) of p-acetamidophenylsulfonyl chloride and 4 24 mg (4.2 mmol) of triethylamine in 20 ml of methylene chloride is stirred for 48 hours at room temperature. The reaction mixture is concentrated under reduced pressure to a foam which is then treated with 10 ml of acetone. The insoluble triethylamine hydrochloride is filtered off, and the filtrate is chromatographed on 160 g of silica gel using acetone as eluent. Fractions 42-86, which by thin layer chromatography have been shown to contain most of the pure product, are collected and concentrated in vacuo to give 1.2 g of the desired product.

NMR (6, CDCl 3 ): 2.06 (3H) s, 2.23 (3H) s, 2.35 (6H) S, 2.70 (2H) S, 3.13 (3H) s, and 7 .6-8.2 (4H) m.

Example 15

ll- acetyl- 4"- deoxy- 4"-( p- cyanophenylsulfonylamino) oleandomycin

A solution of 2.55 g (3.5 mmol) of 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 734 mg (3.65; mmol) of p-cyanophenylsulfonyl chloride and 518 µl (3.75 mmol) of triethylamine in 25 ml of methylene chloride is stirred at room temperature overnight. The solvent is removed,

in vacuo, and the residue is treated with 10 ml of acetone. The insoluble components are filtered off, and the filtrate is chromatographed on 120 g of silica gel using acetone as eluent, and fractions of greater than 10 ml are collected. Fractions 47-83 are combined and concentrated under reduced pressure to give 281 mg of the desired product . -

NMR (6, CDCl 3 ): 2.10 (3H) s, 2.36 (6H) s, 2.71 (2H) d, 3.06 (3H) s, and 7.7-8.4 (4H) m.

Example 16

ll-acetyl-4".rdeoxy-4"-(p-trifluoromethylphenylsulfonylamino)oleandomycin

891 mg (3.65 mmol) of p-trifluoromethylphenylsulfonyl chloride, and the resulting reaction mixture is stirred for 18 hours. The solvent is removed under reduced pressure, and the residue is triturated with 15 ml of acetone. The solids are filtered off, and the filtrate is chromatographed over silica gel to give 287 mg of the desired product.

NMR (6, CDCl 3 ): 2.03 (3H) s, 2.31 (6H) s, 2.63 (2H) d, 3.40 (3H) s,

and 7.15-8.3 (4H) m.

Example 17

ll- ac ety l- 4"- deoxy- 4"-( 2, 2, 2- trifluoroethylsulfonylamino) oleandomycin

A solution of 2.55 g (3.5 mmol) of 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 666 mg (3.65 mmol) of 2,2,2-trifluoroethyl-sulfonyl chloride and 379 mg ( 3.75 mmol) of triethylamine in 25 ml of methylene chloride is stirred for 30 hours at room temperature. An additional 333 mg of sulfonyl chloride and 270 µl of triethylamine are added, and stirring is continued for 4 hours. The solvent is then removed in vacuo,

and the residue is treated with 20 ml of acetone. The solids are filtered off, and the filtrate is chromatographed on 110 mg of silica gel using acetone as eluent and collecting fractions of 10 ml. Fractions 50-80 are mixed and concentrated to give 385 mg of the desired product.

NMR (δ, CDCl 3 ): 2.06 (3H) s, 2.26 (6H) s, 2.60 (2H) d, and 3.36 (3H) s.

By starting with 11-propionyl-4"-deoxy-4"-amino-oleandomycin instead of the 11-acetyl ester and using the method indicated above, the corresponding 11-propionyl-4"-deoxy-4"-(2,2, 2-trifluoroethylsulfonylamino)oleandomycin.

Example 18

11-acetyl-4"-deoxy-4"-(methylsulfonylamino) oleandomycin

A solution of 2.91 g' (4.0 mmol) of 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 467 mg (4.1 mmol) of methylsulfonyl chloride and 424 mg (4.2 mmol) of triethylamine in 25 ml methylene chloride is stirred at room temperature overnight. The solvent is removed under reduced pressure, and the residue is treated with 20 ml of acetone. The triethylamine hydrochloride is filtered, and the filtrate containing the product is chromatographed on 180 g of silica gel using acetone

as eluent, collecting fractions of 6 ml.

Fractions 67-133 are mixed and concentrated in vacuo to give

1.2 g of the desired product.

NMR (6, CDCl 3 ) : 2.06 (3H) s, 2.28 (6H) s, 3.06 (3H) s, 2.61 (2H) d,

and 8.40 (3H) p.

Example 19 11-acetyl-4"-deoxy-4"-(3,4-dichlorophenylsulfonylamino)oleandomycin

11-acetyl-4"-deoxy-4"-amino-oleandomycin (2.9 g, 4.0 mmol),

1.0 g (4.1 mmol) of 3,4-dichlorophenylsulfonyl chloride and 0.57 ml (4.2 mmol) of triethylamine are mixed in 30 ml of methylene chloride, and the resulting solution is stirred at room temperature for 18 hours. The reaction mixture is quenched with 50 ml of water, and the organic phase is washed with a saturated salt solution and dried over sodium sulphate. The solvent is removed in vacuo, and the residue is chromatographed on 150 g of silica gel using acetone as eluent. The fractions containing the product, as detected by thin-layer chromatography,

are collected and concentrated to dryness to give 1.3 g of the desired product.

NMR (6, CDCl 3 ): 2.0 (3H) s, 2.30 (6H) s, 2.60 (2H) d, 3.06 (3H) s

and 7.2-8.1 (3H) m.

Example 20

By following the procedure according to example 19 and ø

starting with the appropriate starting materials, the following compounds are prepared:

Example 21

ll- acetyl- 4"- deoxy- 4"-( 2, 3, 4- trichlorophenylsulfonylamino) oleandomycin

A solution of 2.9 g (4.0 mmol) of 11-acetyl-4"-deoxy-4"-amino-oleandomycin, 1.15 g (4.1 mmol) of 2,3,4-trichlorophenylsulfonyl chloride and 0 .5 7 ml (4.2 mmol) of triethylamine in 30 ml of methylene chloride is stirred at room temperature for 18 hours. The organic layer is washed with water (1 x 50 ml) and a saturated saline solution (1 x 50 ml) and then dried over sodium sulfate. The solvent is removed in vacuo, and the residue is chromatographed on 150 g of silica gel using acetone as solvent, collecting fractions of 7 ml each. Fractions 60-100 are pooled and concentrated to give 800 mg of the desired product.

NMR (6, CDCl 3 ): 2.06 (3H) s, 2.33 (6H) s, 2.63 (2H) d, 3.2 (3H) s

and 7.2-8.2 (2H) m.

By starting with the appropriate starting materials and following the procedure described above, the following compounds are correspondingly prepared: 11-acetyl-4"-deoxy-4"-(3,4,5-trichlorophenylsulfonylamino)oleandomycin; 11-propionyl-4"-deoxy-4"-(2,4,6-trichlorophenylsulfonylamino)-oleandomycin; and 11-acetyl-4"-deoxy-4"-(2,3,5-trichlorophenyl-sulfonylamino)oleandomycin.

Example 22

ll- acetyl- 4"- deoxy- 4"-( 2- hydroxy- 3, 5- dichlorophenylsulfonylamino)-oleandomycin

The procedure according to example 33 is repeated, while

starting with 2.55 g (3.5 mmol) of 11-acetyl-4"-deoxy-4"-amino-. oleandomycin, 954 mg (3.65 mmol) of 2-hydroxy-3,5-dichlorophenylsulfonyl chloride and 518 yl (3.75 mmol) of triethylamine in 25 ml of methylene chloride to give, after chromatography on 220 g of silica gel,

48 3 mg of the desired product.

NMR (6, CDCl 3 /DMS0): 2.03 (3H) s, 2.50 (6H) s, 3.05 (3H) s, and 7.2-7.8 (3H) m.

Example 23

ll- acetyl- 4"- deoxy- 4"-( 3- aminc— 4- chlorophenylsulfonylamino) oleandomycin

A suspension of 500 mg of 10% palladium-on-charcoal in 50 ml of ethyl acetate containing 1.0 g of 11-acetyl-4"-deoxy-4"-(3-nitro-4-chlorophenylsulfonylamino)oleandomycin is shaken in a hydrogen atmosphere at an initial pressure of 3.5 kg/cm 2 at room temperature overnight. The spent catalyst is filtered off, and the solvent is removed

in vacuum. The remaining white foam is chromatographed on 160 g of silica gel using acetone as eluent, collecting fractions of 50 ml each. The fractions containing the product are pooled and concentrated under reduced pressure to give 450 mg of the desired material.

NMR (6, CDCl 3 ): 2.03 (3H) s, 2.33 (6H) s, 2.66 (2H) d, 3.16 (3H) s, and 7.2-8.0 (3H) m.

Under the application of the appropriate; nitro compound according to example 11 is prepared in a similar way: 11-acetyl-4"-deoxy-4"-(m-aminophenylsulfonylamino)oleandomycin NMR (6, CDCl 3 ): 2.03 (3H) s, 2.30 (6H) s, 2.63 (2H) d, 3.10 (3H) s and 7.0-7.8 (4H) m; and 11- acetyl- 4"- deoxy- 4"-( p- aminophenylsulfonylamino) oleandomycin NMR (6, CDCl 3 ): 2.06 (3H) s, 2.31 (6H) s, 3.02 (3H) s and 6.4-7.8 (4H) dd.

Example 24

ll- acetyl- 4"- deoxy- 4"-( 3- methyl 1- 2- thienylsulfonylamino) oleandomycin

To 100 g (0.13 mol) of 11-acetyl-4"-deoxy-4"-amino-oleandomycin in 900 ml of methylene chloride is added 59 3 ml of triethylamine,

and the solution is stirred for 10 minutes. 3-Methyl-2-thienylsulfonyl chloride (41.9 g, 0.213 mol) in 300 mL of methylene chloride is then added dropwise over a period of 1 hour, and the reaction mixture is stirred at room temperature for 48 hours. The reaction mixture is added to 2 liters of water, and the organic layer is separated, washed successively with water (2 x 250 ml) and a salt solution (1 x 250 ml) and dried over sodium sulfate. The solvent is removed in vacuo, and the residue is chromatographed on a 105 cm x 6.5 cm column containing 1.5 kg of silica gel. The product, which is eluted with acetone, is collected from the 2.3 liter to 6 liter eluate fractions. The fractions are pooled and the solvent is removed under reduced pressure to give a foam. Treatment of the remaining foam with diethyl ether yields 66.4 g of the desired product, m.p. 184-185.5°C.

NMR (<5, CDCl 3 ) : 2.04 (3H) s, 2.41 (6H) s, 2.46 (3H) s, 2.62 (2H) m, 3.02 (3H) s, 6, 84 and 7.32 (2H).

To 2 g of the above-mentioned free base in 15 ml of ethyl acetate is added 0.12 ml of phosphoric acid, and the resulting solution is allowed to stir at room temperature. After 20 minutes crystals begin to form, and after 2 hours these are filtered, washed with ethyl acetate and dried to give 1.3 g of 11-acetyl-4"-deoxy-4"-(3-methyl-2-thienylsulfonylamino)oleandomycin -phosphate.

NMR (<S, CD30D) : 2.01 (3H) s, 2.45 (3H) s, 2.56 (2H) m, 2.83 (6H) s, 3.0 (3H) s, 6, 88 and 7.42 (2H).

Example 25

The procedure according to example 24 is repeated, while

starting with the appropriate sulfonyl chloride and 11-acetyl-4"-deoxy-4"-amino-oleandomycin to prepare the following related compounds:

Example 26

ll- acetyl- 4"- deoxy- 4"-( 5- carbomethoxy- 2- pyrrylsulfonylamino)- oleandomycin

A solution of 2.96 g (0.0041 mol) 11-acetyl-4"-deoxy-4"-amino-oleandomycin and 0.62 ml of triethylamine in 50 ml of dry methylene chloride cooled to ice-bath temperature is treated in portions with 1.0 g (0.0044 mol) 2-carbomethoxy-5-pyrrylsulfonyl chloride. The reaction mixture is heated to room temperature and stirred for 3.5 hours, and then poured into 200 ml of water. The pH of the aqueous layer is adjusted to 9.5 with 1N aqueous sodium hydroxide, and the methylene chloride layer is separated, washed successively with water and saturated salt solution and dried over sodium sulfate. Removal of the solvent under reduced pressure gives 3.8 g of the crude product as a white foam.

Said foam is then chromatographed on a silica gel column 3.25 cm x 38 cm using acetone as eluent. Fractions 40-220, which make up approx. 10-12 ml each, collect and mix. Removal of the solvent in vacuo gives 3.4 g of the desired product as a white foam.

NMR (6, CDCl 3 ): 2.05 (3H) s, 2.58 (6H) s, 2.67 (2H) m, 3.25 (3H) s, 3.90 (3H) s, 7.20 (IB) m and 7.52' (1H) m.

Example 27

The procedure of Example 26 is repeated, starting with the appropriate sulfonyl chloride and 11-acetyl-4"-deoxy-4"-amino-oleandomycin, to prepare the following analogues:

Example 27

4"-deoxy-4"-(p-chlorophenylsulfonylamino)oleandomycin

A solution of 3.0 g of 4"-deoxy-4"-amino-oleandomycin, 8.65 mg of p-chlorophenylsulfonyl chloride and 424 mg of triethylamine in 25 ml of methylene chloride is stirred at room temperature overnight. The solvent is removed in vacuo, and the residue is treated with 20 ml of acetone. The insoluble triethylamine hydrochloride is filtered, and the filtrate is chromatographed on 180 g of silica gel using acetone as eluent, fractions of 50 ml being collected. Fractions 18-27 are combined and concentrated under reduced pressure to give 1.10 g of the desired product.

NMR (6, CDCl 3 ): 2.33 (6H), 2.83 (2H) d, 3.06 (3H) s, and 7.2-8.4 (4H) m.

Example 28

4"-deoxy-4"-(p-toluenesulfonylamino)oleandomycin

By a procedure similar to that described in

example 27, 30 g (4.0 mmol) of 4"-deoxy-4"-amino-oleandomycin, 782 mg (4.1 mmol) of p-toluenesulfonyl chloride and 424 mg (4.2 mmol) of triethylamine are stirred in 25 ml of methylene chloride at ambient temperature at night

above. During work-up, the crude product is chromatographed on 180 g • .-silica gel, collecting fractions of 10 ml. Fractions 90-148 were pooled and concentrated to dryness to give 1.4 g of the desired product.

NMR (6, CDCl 3 ): 2.33 (6H) s, 2.46 (3H) s, 2.83 (2H) d, 3.10 (3H) s, and 7.10-8.0 (4H) m.

By a similar method, 4"-deoxy-4"-(2-thienylsulfonylamino)oleandomycin is also produced.

NMR (6, CDCl 3 ) : 2.29 (6H) s, 2.88 (2H) m, 3.2 (3H) s, 5.6 (1H) m

and 7.33 (3H) m.

Example 29

ll- acetyl- 4"- deoxy- 4"-( 2- thienylsulfonylamino) oleandomycin- phosphate

To a solution of 15.0 g of 11-acetyl-4"-deoxy-4"-(2-thienylsulfonylamino)oleandomycin in 100 ml of ethyl acetate is added

1.0 ml phosphoric acid. The resulting suspension is stirred for 4 hours at room temperature. The solids are filtered off, washed with ethyl acetate and dried to give 12.5 g of the desired salt,

sm.p. 168°C (dec.).

11-Acetyl-4"-deoxy-4"-(3-methyl-2-thienyl sulfonylamino)oleandomycin phosphate is prepared in a similar manner,

sm.p. 184-188°C and 11-acetyl-4"-deoxy-4"-(p-chlorophenylsulfonylamino)oleandomycin phosphate, m.p. 204-205°C.

MANUFACTURE A

4"- deoxy- 4"- oxo- oleandomycins

I. 11-acetyl-4"-deoxy-4"-oxo-oleandomycin

a. 11, 2'- diacetyl- 4"- deoxy- 4"- oxo- oleandomycin

To 4.5 g of N-chlorosuccinimide, 50 ml of benzene and 150 ml of toluene in a dry flask equipped with a magnetic stirrer and nitrogen inlet and cooled to -5°C, add 3.36 ml of dimethyl sulphide. After stirring at 0°C for 20 minutes, the contents are cooled to -25°C and treated with .50 g of 11,2'-diacetyl-oleandomycin in 100 ml of toluene. Cooling and stirring are continued for 2 hours followed by the addition of 4.73 ml of triethylamine. The reaction mixture is stirred at 0°C for 15 minutes and then poured into 500 ml of water. The pH is adjusted to -9.5 with IN aqueous sodium hydroxide, and the organic layer is separated,

washed with water and saline solution and dried over sodium sulfate. Removal of the solvent in vacuo gives 4.9 g of the desired product as a foam.

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

b. 11-acetyl-4"-deoxy-4"-oxo-oleandomycin

A solution of 4.0 g of 11,2'-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

the product as a foam. A diethyl ether solution of the residue gives, on treatment with hexane, 2.6 g of the product as a white solid, m.p. 112-117°C.

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

By using 11,2<1->dipropionyl-4"-deoxy-4"-oxo-oleandomycin or 11-propionyl-2"-acetyl-4"-deoxy-4"-oxo-oleandomycin by the method described above, the corresponding 11-propionyl-4"-deoxy-4"-oxo-oleandomycin is obtained.

II. 4"- deoxy- 4"- oxo- oleandomycin

a. 2'- acetyl- 4"- 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 cooled to -5°C and maintained under a nitrogen atmosphere. After stirring at 0°C for 20 minutes, the mixture is cooled to -25°C, and 1.46 g of 2'-acetyloleandomycin and 15 ml of toluene are added. Stirring is continued for 2 hours at -20°C followed by the addition of 0.46 ml of triethylamine. The reaction mixture is kept at -20°C for an additional 5 minutes and then warmed to 0°C. The mixture is poured with stirring into 50 ml of water and 50 ml of ethyl acetate. The pH of the aqueous mixture

adjusted to 9.5 by adding aqueous sodium hydroxide. resolution. 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, as at two

recrystallization from methylene chloride-diethyl ether yields 212 mg of the pure product, m.p. 183-185.5°C.

Analysis: Calculated for c37H6i°13N: c 61.1, H 8.5, N 1.9.

Found: C 60.9, H 8.4, N 1.9.

NMR (6, CDCl 3 ): 5.60 (1H) m, 3.50 (3H) s, 2.73 (2H) m, 2.23 (6H) s and 2.03 (3H) s.

b. 4"- deoxy- 4"- oxo- oleandomycin

One 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, 9 37 mg.

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

MANUFACTURE B

4"- deoxy- 4"- amino- oleandomycins

I. 11-Acetyl-4"-deoxy-4"-amino-oleandomycin

To a suspension of 10 g of 10% palladium-on-charcoal i

21.2 g of ammonium acetate are added to 100 ml of methanol, and the resulting slurry is treated with a solution of 20 g of 11-acetyl-4"-deoxy-4"-oxo-oleandomycin in 100 ml of the same solvent. The suspension is shaken at room temperature in a hydrogen atmosphere at

an initial pressure of 3.5 kg/cm 2 . After 1.5 hours, the catalyst is filtered off, and the filtrate is added to a mixture of 1200 ml of water and 500 ml of chloroform with stirring. The pH is regulated from 6.4 to 4.5, and the organic layer is separated. After a further extraction with 500 ml of chloroform, the aqueous layer is treated with 500 ml of ethyl acetate, and the pH is adjusted to 9.5 with IN sodium hydroxide. The ethyl acetate layer is separated, and the aqueous layer is extracted again with ethyl acetate. The ethyl acetate extracts are collected, dried over sodium sulfate and concentrated to a yellow foam (18.6 g), which upon crystallization from diisopropyl ether gives 6.85 g of purified product,

sm.p. 157.5-160°C.

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

The other epimer, which exists in the raw foam in an amount of 20-25%, is obtained by gradual concentration and filtration of the mother liquors.

Similarly, by starting with 11-propionyl-4"-deoxy-4"-oxo-oleandomycin by the method described above, 11-propionyl-4"-deoxy-4"-amino-oleandomycin is obtained.

II. 4"- deoxy- 4"- amino- oleandomycin

A solution of 20 g of 2'-acetyl-4"-deoxy-4"-oxo-oleandomycin in 125 ml of methanol, after stirring at room temperature overnight, is treated with 21.2 g of ammonium acetate. 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 2 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 in the aqueous layer is regulated

to 8.25. The diethyl ether and ethyl acetate extracts obtained at this pH are also set aside and the pH is raised to 9.9. diethyl ether-

and the ethyl acetate extracts from this pH are combined, washed successively with water (1x) and a saturated saline solution and dried over sodium sulfate. The final extracts, taken at pH 9.9, are concentrated to a foam and chromatographed on 160 g of silica gel using chloroform as the developing agent and the first eluent.

After 11 fractions of 12 ml per fraction, the eluent is changed to 5% methanol - 95% chloroform. For fraction 370, the eluent is changed to 10% methanol - 90% chloroform, and for fraction 440, 15% methanol - 85% chloroform is used. Fractions 85-260 are combined and concentrated in vacuo to dryness to give 2.44 g of the desired product.

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

Claims (2)

1. Analogous method for the preparation of a therapeutically active compound of the general formula: and pharmaceutically acceptable acid addition salts thereof,.. hydr R ler • • alkyl med. 1 to 3 carbon atoms; pyridyl';. 1 ,.1,1-trifluqrety 1;:,; '.' phenyl; monosubstituted phenyl. where the substitute.,.ef.-rfluqf'M:kibr'r v'-•: ^' bromo, iodo, hydroxy, methoxy, cyano, carboxamido; -ri i tro, amino, carbomethoxy, carbobenzyloxy, carboxy, trifluoromethyl, alkyl of 1 to 4 carbon atoms or acetamido; disubstituted t-phenyl wherein the substituents are each chloro, nitro, amino, methoxy or methyl; trichlorophenyl; hydroxydichlorophenyl; benzyl; naphthyl; thienyl; chlorothienyl; 2-acetamido-5-thiazolyl; 2-acetamido-4-methyl-5-thiazolyl; 2-benzimidazolyl; dimethyl-2-pyrimidinyl; pyrryl; furyl; mono-substituted thienyl, pyrryl or thienyl wherein each substituent is carbomethoxy or alkyl of 1 to 2 carbon atoms; or 1-methyl-5-carbomethoxy-3-pyrryl; and is hydrogen or alkanoyl with 2 to 3 carbon atoms, characterized in that a compound with the formula: is reacted with one mole of a sulfonyl halide with the formula RS02W wherein W is a halogen atom, and in the presence of an acid scavenger in a reaction-inert solvent at ambient temperature, provided that when R 1 is hydrogen, R 2 is phenyl; thienyl; monosubstituted phenyl wherein the substituent is chloro, fluoro, methyl, methoxy or trifluoromethyl; or alkyl substituted thienyl where the alkyl group contains 1 to 2 carbon atoms. 2. Process as stated in claim 1, characterized in that starting materials are used where R 1 is acetyl and R is 2-thienyl, 3-thienyl or 3-methyl-2-thienyl.