IL27461A - 7-deoxylincomycin,analogs and isomers thereof and their preparation - Google Patents

Description

anaam n+ftv o»¾m»K¾ o'aiVan .ι»3»ρτρ3» »ορΐκη·τ-7 7-DEOXTl I NCOMYCI N. ANALOGS AND ISOMERS THEREOF AND THEIR PREPARATION This invention relates to novel compounds and to processes for preparing them, and is particularly directed to 7—dooxylineomycin, and analogs and isomers hereof and to processes whereby they and like compounds are produced.

The novel compounds of the invention ca be represented by the following structural formulas wherein H s a y not mo e thaa 20 carbon a oms nd lie is hydrogen or the acyl radical of a 4-^substituted-L—2-pyrroiidine- carboxylic acid of the formula wherein li^ and are selected from the grou consisting of alkylidine and F*j alky7x of not more than 20 carbon atoms, cycloalkylide e and Ii cycloalkyl of from 3 to not more than 8 carbon atoms, aralkylidene and lij aralkyl of not more ■ftan 12 carbon atoms; E is a protective hydr car ony1- oxy-carbonyl group, diphenyl (p-taethoxyphenyl) methyl , bis(p-methoxyphenyl)phonylmethyl, benzyl, or p-nitrobe.'azyl ; and is selected iro the group consisting of h dro en and U Examples of alkyl of not more than 20 carborv atoms (R, H i, and HR2) are methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl , tri-decyl , tetradecyl, pentadecyl, hexadecyl , heptadecyl, octa-decyl, nonadecyl, and eicosyl and the isomeric forms thereof. Examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclo-pentyl, cyclohexyl, cycloheptyl, cyclooctyl, 2-methyl cycl o-pentyl, 2, 3-dimethyl cyclobutyl , 2-methyl cyclobutyl , and 2-cyclopentyl propyl . Examples of aralkyl are benzyl, phen-ethyl, 3-phenyl propyl , and 1-naphthylmethyl . Examples of alkyl idene, cyclpa 1 kyl i dene, and aralkylidene groups (Rj, and R2) include methyl ene, ethyl i dene, propyl idene, butyl idene, pentyl idene, hexylJdene, heptyl idene, octyl idene, nonyl idene, decyl idene, undecyl i dene, dodecyl i dene, tri decyl i dene, tetradecyl idene, pentadecyl idene, hexadecyl i dene, heptadecyl i dene, octadecyl idene, nonadecyl i dene, ei cosyl t dene, and the isomeri forms thereof, cyclopropyl i dene, cyclobutyl idene, cyclopentyl idene, cycl ohexyl i dene, cycl oheptyl i dene, cycl ooctyl i dene, 2-cycl opropyl ethyl i dene, 3-cyclopentyl propyl i dene, benzyl -idene, 2-phenyl ethyl i dene, 3-phenylpropyl idene, and 1-naphthylmethyl ene.

The novel compounds of the invention, Formula I, as well as other related compounds, ea -be- prepared by replacing by halogen, the 7-hydroxy of a compound of the Formula I I to form a 7- halo compound of Formula III and deshalogenati ng the latter to form a compound of Formula I.

The deshalogenatlon Is effected advantageously by mixing the starting compound of Formula III with zinc dust and heating.

In the process Ac and R can be any radical which is non- reactive with zinc dust, but for preparing compounds of the Invention Ac and ft are as given above. For example, when a compound of Formula IMA (Ac in Formula III is that o the acid of Formula A) is used as the starting compound, a novel compound of Formula IA is obtained. When compound IA is hydrogenated with a catalyst effective to saturate an olefinic double bond, or the starting compound (Formula MIA) is similarly hydrogenated and then deshalogenated, a compound of Formula IB is obtained as a mixture of c? s and trans epimers according to the formulas IVA and I B which, if desired, can be separated by counter current distribution or chromatography.

When Rs (HRa) In Formulas B, IB, I IB, and 1MB is hydrogen, it can be replaced by suitable alkylation or like procedure. Advantageously, this replacement is effected by reacting the compound according to Formula B, IB* MB, or 1MB, wherein R3 is hydrogen with an oxo compound (an alde hyde or a ketone) and hydrogenating the resulting adduct with a catalyst effective to saturate an olefinic double bond. Either platinum or palladium can be used as the catalyst. Suitable oxo compounds have the formula R4R5CO where R4R5C= is the same as R2 given above. Examples of suitable oxo compounds are formaldehyde, acetal dehyde, propionaldehyde, butyral dehyde, acetone, ispbutyl methyl ketone, benzal dehyde, phenylacetaldehyde, hydroci nnamal dehyde, acetophenone, pro-piophenone, butyrophen ne, 2-methyl - -pheny 1 -2-butanone, 2-methyl -5-phenyl -3-pentanone, 3-cyclopentanepropionaldehyde, cycl ohexaneacetal dehyde, cycl oheptanecarboxal dehyde, 2,2-dimethyl cycl opropyl acetal dehyde, 2,2-dimethyl cyclopropyl methyl ketone, cyclopentyl methyl ketone, cyclobutyl methyl ketone, cycl obutanone, cycl ohexanone, 4-methyl cycl ohexanone, and the 1 i ke.

The starting compounds of Formula M ?Tre prepared by acylati d of the formula wherein R is as given above, with a -substi tuted-L-2-pyrrol idinecarboxylic acid of Formula A or B. This acylation was and like acyiations referred to herein -caw-be effected by procedures already well known in the art for acylating amino The starting acid of Formula A can be prepared by a 4-o wherein Z is a protective hydrocarbyloxycarbonyl group which is removable by hydrogenol ys i s , trityl, i.e., tr i phenylmethyl , di phenyl (p-methoxyphenyl )methyl , bi s- (p-methoxyphenyl )phenyl -methyl, benzyl, or p-ni trobenzyl , with a Wittig agent, e.g., an al kyl i denetr i phenyl phosphorane [see e.g., Wittig et al., Ber., 87, 13^8 (195*0; Trippett, Quarterly Reviews, XVII, No. 4, p. 4o6 (1963)], Examples of hydrocarbyloxycarbonyl groups (Z) are tertiary-butoxycarbonyl ; benzyloxycarbonyl groups of the formula wherein X is hydrogen, nitro, methoxy, chloro, or bromo, for example, ca rbobenzoxy, p-ni trocarbobenzoxy, p-bromo-, and p-chlorocarbobenzoxy; and phenyt¾xycarbonyl groups of the formula wherein Xx is hydrogen, alJyl, or alkyl of not more than 4 carbon atoms, such as p heny4 >xy car bony 1 , p- tol yl oxycarbonyl , p-ethyl pheny-roxycarbonyl , and p-al 1 yl pheriyioxycarbonyl and the I i ke.

In carrying out this process the 4-oxo-L-2-pyrrol i -d i necarboxyl i c acid (Formula C) is added to a freshly prepared Wittig reagent. The Wittig reagents herein used can be generally represented by the following formula: i=P(CeHs)3 wherein t is as given above. These Wittig reagents are prepared by reacting an alkyl, cycloalkyl, or aralkyltri-phenyl phosphoni urn hal ide with a base such as sodamide, or sodium or potassium hydride, or the sodium or potassium metalate of dimethyl sulfoxide and the like. For example, the el imi nat ion of hydrogen hal ide from al kyl triphenylphos-phonium hal ide, produces al kyl i denetri phenyl phosphorane.

[The preparation of phosphoranes is discussed in detail by Trippett, Quart. Rev. XVII, No. 4, p. 4o6 (1963)]. The reaction is generally carried out in an organic solvent, such as benzene, toluene, ether, dimethylsul foxide, tetra- o hydrofuran, or the like, at temperatures between 10 C. and the reflux temperature of the reaction mixture. The thus-obtained product, a 4-al kyl i dene-, 4-cycl oal kyl i dene-, or 4-aral kyl i dene-l-protected-L-prol i ne whi ch has the following formula: z is recovered from the reaction mixture in a conventional manner, generally by extraction from aqueous solutions of the reaction mixture. The crude product can be purified by conventional means, such as recrystall ization, chromatography, or formation and recrystal 1 i zat ion of easily formed derivatives such as amine salts of the amino acid, e.g., the dicyclohexylami e saltj and the l ike, and liber ating the amino acids from such compounds. By hydrogenating an acid of Formula D in the presence of a catalyst, e.g., platinum, which is effective to saturate a double bond, but which is ineffective to effect hydrogenol ys i s, a compound of the following formula: is obtained. Platinum deposited on a carrier, e.g., carbon or an anion exchange res i n 1 i ke Dowex-1, a cross- linked polystyrene trimethyl benzylammoni urn resin in the hydroxide cycle is suitable. If desired, the starting compounds of Formula V can be acylated with acids of Formula C, D, or E to form compounds IIC, I ID, and I IE, respectively. Compound IIC can then be converted to compound I ID by .treatment with a Wittig reagent and compound I ID hydrogenated to compound I IE by the procedures given above. The hydrogenat ion, both of the acid D and the acylate I ID, gives a mixture of cis and trans epimers which, if desired, can be separated by counter current distribution or chromatography. The starting acids of Formula B in which R3 is hydrogen are obtained when an acid of Formula D or E is subjected to hydrogenol ys is over a palladium catalyst, e.g., palladium on carbon. Likewise, compounds of Formula I ID and I IE are converted to compounds of Formula MB in which R3 is hydrogen by the same process. The starting acids of Formula B in which R3 is hydrogen as well as compounds of Formula MB in which R3 is hydrogen can be converted respectively to compounds of Formulas B and I IB in which R3 is HR2 by the procedures given above.

The starting acids of Formula A are obtained by treating an acid of Formula D with hydrogen bromide in acetic acid to remove the Z group and then replacing the N-hydrogen with an H 2 group by the procedure given above. Compounds of Formula I ID and I IE are converted to compounds of Formula t I A and MB, respectively, by the same process.

Some of the starting compounds of Formula II are obtained b iosyntheti cal 1 y. Lincomycin, methyl 6,8-dideoxy-6- (trans*!-methyl - -propyl -L-2-pyrrol idi necarboxamido)-l-thio-D-erythro-a-D-gal acto-octopyranos ide, is obtained as an elaboration product of a 1 i ncomyci n-produci ng actinomycete according to U. S. Patent 5*086,912. It has the following structural formula: wherein R and R3 are methyl and iH is propyl. Lincomycin B, methyl 6,8-dideoxy-6- (trans-l-methyl - -ethyl -L-2-pyrrol i -di necarboxami do)-l- thio-D-erythro-g-D-galacto-octopyranos ide (Formula VI wherein R and R3 are methyl and -R^ is ethyl) also is an elaboration product of the same microorganism when cultured according to the procedure given in U. S. Patent ,086,912. Lincomycin C, ethyl 6,8-di deoxy-6- (trans -1-methy I -4-propyl -L-2-pyr rol i d i necarboxami do) -1- thi o-D-erythro-a-D- gal acto-octopyranosi de (Formula VI wherein R is ethyl, - iH is propyl, and R3 is methyl) is obtained when the process of U. S. Patent 3,086,912 is carried out in the presence of added ethionine. Lincomycin D, methyl 6,8-di deoxy-6- (trans- -propyl -l-2-pyrrol i d i necarboxami do)-l-thi o-D-erythro-a-D-ga 1 acto-octopy ranos i de (Formula VI wherein R is methyl, -R4H is propyl, and R3 is hydrogen) is obtained when the fermentation of U. S. Patent 3,086,912 is carried out in the presence of added a-MTL, methyl 6 -ami no-6,8-d ideoxy-D-erythro-1-thi o- g-D -gal acto-octopy ranos ide, a compound obtained by the hydraz i nol ys i s of lincomycin. Methyl 6,8-di deoxy-6- (trans- -ethyl -L-2-pyrrol id i necarboxami do) -1- thi o-D-erythro-g-D-galacto-octopyranoside (Formula VI wherein R is methyl, - iH is ethyl and R3 is hydrogen) is also produced when a-MTL is added to the fermentation of U. S. Patent 3, 086,912. Similarly, lincomycin K, ethyl 6,8-di deoxy-6- (trans -4- propyl -L-2-pyrrol i -di necarboxami do) -1-th? o-D-erythro- -D- gal acto-octopy ranos i de (Formula VI wherein R is ethyl, -RxH is propyl, and R3 is hydrogen) is produced when the fermentation of U. S. Patent 3,086,912 is carried out in the presence of added g-ETL, ethyl 6-ami no-6,8-d i deoxy-1- thi o-D-erythro-g-D- gal acto-octopy rano-side, a compound obtained by the hydraz i nol ys i s of lincomycin C. Ethyl 6,8-di deoxy-6- (jtrans- -ethyl -L-2-pyrrol id! necarbox-ami'do)-!- thio-D-erythro-g-D-galacto-octopyranos ide (Formula VI wherein R is ethyl. ,-R4H is ethyl, and R3 is hydrogen) is also obtained when g-ETL is added to the fermentation of U.S. Patent ,3, 086,912. The above-described N-desmethyl products which are obtained when g-MTL and g-ETL are added to the fermentation process of U. S. Patent 3,086,912 are examples of compound MB wherein R3 is hydrogen. By the procedure described above replacement of the N- hydrogens produces compounds of Formula MB wherein R3 equals N2 e.g. , methyl 6,8-di deoxy-6- (trans-1-ethyl -4-propyl -L-2-pyrrol Idl necarbox-ami do)-l-thio-D-erythro-o D-galacto-octop!yranoside, ethyl 6,8-dl deoxy-6- (trans-1-methyl -4-ethyl -L-2-pyrrol Idinecarbox-amldo)-l-thio-D-erythro-a-D-galacto-octopyranoside, ethyl 6,8-di deoxy-6- (trans-l-ethyl -4-ethyl -L-2-pyrrbl Idlriecarbox-amIdo)-l-thlo-D-erythro-a-D-gaiacto-octopyranoslde, and methyl 6,8-di deoxy-6- (trans-l-ethyl -4-ethyl -L-2-pyrrol id! necarboxamf do)-l-thlo-D-erythro-a-D-galacto-octopyranoslde.

As the blosynthet ί c-produced ti neomycins, as -well as the amino sugars derived therefrom, are either methyl or ethyl thlogtycos ides, tt is sometimes desirable to convert them to higher or lower glycosides. It Is sometimes desirable to convert any of the compounds of Formulas 1,11, III, or V to higher or lower glycosides. This can effectively be accomplished by reacting the compound to be converted with a mercaptan of the formula ReSH wherein Re Is art alkyl group of not more than 20 carbon atoms, but one different from R and cyclizing the resulting dlmercaptal. For example, compounds of Formula I, II* and III on reaction with a mercaptan of Formula ReSH produce dl thioacetals of the formula wherein X is hydrogen, hydroxy or halogen, which on treatment with acid or on heating, either in the presence of acid or not, are recyclized to a compound of the following formul a : The process advantageously Is applied to any of the starting products of Formula II, i.e.. MA, MB, IIC, I ID, and ME. The resulting products can be subjected to hydraz i nol ys i s to form compounds of the following formula: OH which can be N-acylated as described above with acids of Formula A, B, C, D, and E to provide compounds according to Formula XII wherein X is hydroxy. The process can also be applied to the starting compounds of Formula V. For example, a-MTL on treatment with ethyl mercaptan followed by cycliza-tion as described above is converted to -ETL.

An alternative process for making compounds of Formula XII or Formula XM I is to brominate the starting material (Formulas 1, II, III, or V) and then react the product with a mercaptan according to the following sequence: The starting compound, XIV, Is dissolved in water as a soluble salt, e.g., the hydrochloride, and bromine added o with cooling advantageously to between about -10 and 20 C.

It is sufficient if the aqueous solution is cooled to about o 0-5 C. and the bromine added dropwise. The stoichiometric amount of bromine is 1 mole for each mole of starting compound, though more or less can be used. Advantageously a slight excess, say from 5 to 20$ excess of bromine is used. The bromine initially replaces the RS- group and the resulting intermediate hydrolyzes to the sugar in which the pyranose form XVa is in equilibrium with the aldose form XVb. In the presence of acid, e.g., hydrochloric acid or other strong non-oxidizing acid such as p-tol uenesul foni c acid and sulfonic acid type anion exchange resins, the mer-captan ReSH reacts wi th the sugar XV to form the thioglyco-side XVI. Concomitantly some di thioacetal may be formed which after separation can be cyclized as described above to form more of the desired thfoglycoside XVI.

The 7-hydroxy group of the starting compounds of Formula I I I can be replaced by chlorine by heating with thionyl chloride. In the process, Ac and can be any radical which is non-reactive with thionyl chloride but for preparing compounds of the invention, Ac and R are as given above. All that is necessary, however, to effect the substitution of the 7-hydroxy by chlorine is to mix the starting compound of Formula I I or V, advantageously in the form of an acid-addition salt, e.g., the hydrochloride, with thionyl chloride, advantageously in the presence of an inert solvent, with mild heating, advantageously at reflux temperature, until the desired substitution of the 7-hydroxy group by chlorine is effected. Advantageously, the reaction is carried out in an inert atmosphere, e.g., under nitrogen. Carbon tetrachloride can be used effectively as the solvent vehicle but other inert solvents such as chloroform, methylene chloride, ethylene chloride, ether, benzene, and the l ike can be used. A satisfactory procedure is to stir the reaction mixture at room temperature for a considerable period, say from about 1 to l8 hours or as long as necessary to obtain a reasonably clear solution and then to raise the o temperature to between about 50 and 100 C, for example, to o the reflux temperature (77 C. for carbon tetrachloride).

After the reaction is complete, usually after heating at re-flux for about 1 to 5 hours, the reaction mixture is al lowed to cool, advantageously under nitrogen. Any material that separates on the cooling is collected and dried. The solvent is removed by vacuum distillation at a pot temperature advan- o tageously less than about 35 C. and the material which pre-cipitates is collected and dried and treated with ethanol to convert any residual sulfite intermediates to the desired product. The collected material can then be further purified by solvent extraction and/or recrystal 1 ization and can be recovered either as the free base or an acid addition salt.

The proportions of the reagents can be varied widely.

Stoichfometrical ly, however, at least 3 moles of thionyl chloride is required for each mole of starting compound. Any larger amount can be used but ordinarily it is not necessary or desirable to use more than about a 10 fold excess. Advan-tageously, an excess of about 2 to 3 fold is used. The amount of solvent is not critical and can be varied widely in accordance with the practices in the art. Ordinarily from about 15 to about 30 volumes of solvent for each part of solid starting compound will suffice. The proportion of solvent to thionyl chloride, however, is important because of the solubility of the product in thionyl chloride. If the ratio of solvent to thionyl chloride (v/v) is high, the desired product precipitates on cooling of the reaction mixture and the work up of the product is simplified. For example, with carbon tetrachloride a mixture of products precipitates directly on cooling the reaction mixture if the v/v proportion of carbon tetrachloride to thionyl chloride is kept above about 10 to 1.

The replacement of the 7-hydroxy group by halogen can be effected by mixing the starting compound of Formula I I with Rydon reagent and heating. In the process Ac and R can be any radical which is non-reactive with Rydon reagent, but for preparing compounds of the invention Ac and R are as given above.

The mechanism by which Rydon reagent effects the sub-stitution of the 7-hydroxy by halogen is not fully understood.

It is believed, however, tha the mechanism is suc that a change in configuration results. Thus, a 7-hydroxy compound of the D-erythro configuration would yield a 7-halo compound of the L-threo configuration.

Rydon reagents are formed by the addition of halogen to trl phenyl phosphi ne or tri phenyl phosphi te or addition of an alkyl halide to trfphenylphosphi te and can be represented by the formulas: wherein X is halogen, e.g., chlorine, bromine, and iodine. ydon et al., J. Chem. Soc. 2224· (1953); Ibid, 228l (195 ); Ibid, 30 3 (1956).

The Rydon reagent can be formed in situ by addition of halogen or methyl halide to a solution of the triphenyl-phosphine or t r i phenyl phosphi te in an inert solvent such as acetonitrile or d imethyl formami de, or it can be isolated as a separate entity. In either case the reaction with the l incomycin or related compounds is effected by contacting the Rydon reagent therewith in an inert solvent, e.g., acetonitri le or dimethyl formami de, unti l the desired substitution of the 7-hydroxy is obtained. The reaction takes place at ordinary temperature, though gentle heating can be effected if desired. Advantageously the temperature is main- o o tained between about 20 C. and about 55 C. The product can be recovered from the reaction mixture by wel l known techniques such as filtration, solvent extraction, etc. The reaction mixture advantageously is heated with methanol to destroy any excess Rydon reagent, filtered to remove any sol id such as t ri phenyl phosphi ne oxide, formed in the reaction, and then treated to recover the product. The methanol can be added either before or after the filtration. Advantageously the treated and filtered reaction mixture is evaporated to dryness and purified by solvent extraction and/or chromatography.

The compounds of Formulas I A, IB, I I A, MB, and V exist either in the protonated or non-protonated forms according to the pH of the environment. When the protonated form is intended the compound is qual ified as an acid-addition salt and when the non-protonated form is intended it is qual ified as the free base. The free bases can be converted to stable acid-addition salts by neutralizing the free base with the appropriate acid to below about pH 7.0, and advantageously to about pH 2 to pH 6. Suitable acids for this purpose in-elude hydrochloric, sulfuric, phosphoric, thiocyanic, fluo-silicic, hexaf luoroarsenlc, hexaf 1 uorophosphor i c, acetic, succinic, citric, lactic, maleic, fumaric, pamoic, chol ic, palmitic, mucic, camphoric, glutaric, glycol ic, phthal ic, tartaric, lauric, stearic, salicyl ic, 3-phenyl sa 1 i cy 1 i c, 5-phenylsal icyl ic, 3-methyl gl utar i c, orthosul fobenzoi c, cyclohexanesul famic, cycl opentaneprop ionic, 1,2-cyclohexane- dicarboxyl ?c, -cyclohexanecarboxyl i c, octadecenyl succi n i c, octenyl succi n? c, methanesul foni c, benzenesul foni c, hel ianthic, Reinecke's, dimethyl di thiocarbamic, cyclohexyl sul fami c, hexa-decyl sul famic, octadecyl sul fami c, sorbic, monochl oroacet i c, undecyl enic, 1 -hyd roxyazobenzene- - sul foni c, octodecyl sul f ur i c, picric, benzoic, cinnamlc, and like acids.

The acid-addi tipn salts can be used for the same purposes as the free base or they can be employed to upgrade the same. For example, the free base can be converted to a water- i nsol ubl e salt, such as. the picrate, which can be subjected to purification procedures, for example, solvent extractions and washings, chromatography, fractional l iquid-liquid extractions, and crystall ization and then used to regenerate the free base form by treatment with alkali or to make a different salt by metathesis. Or the free base can be converted to a water-soluble salt, such as the hydrochloride or sulfate, and the aqueous solution of the salt extracted with various water- immi sci bl e solvents before regenerating the free base form by treatment of the thus-extracted acid solution or converted to another salt by metathesis. The free bases of Formulas IA, IB, MA, MB, I MA, 1 MB, and V can be used as buffers or as antacids. The compounds of Formulas I, M, II I and V react with isocyanates to form urethanes and can be used to modify polyurethane resins. The long chain compounds, i.e., where HRs is alkyl of from 8 carbon atoms up, have surface active properties and can be used as wetting and emulsifying agents. The thiocyanic acid addition salt when condensed with formaldehyde forms resinous materials useful as pickling inhibitors according to U. S.

Patents 2,425,320 and 2,βθ6,155. The free bases also make good vehicles for toxic acids. For example, the fluosilicic acid addition salts are useful as mothproofing agents according to U. S. Patents 1,915,32 and 2,075,359 and the hexa- f 1 uoroarseni c acid and hexaf 1 uorophosphor i c acid addition salts are useful as parasiticides according to U. S. Patents 3,122,536 and 3,122,552.

The close analogues of l incomycin, i.e., where - tH is ci s or trans alkyl of not more than 8 carbon atoms; R3 is methyl or ethyl; R is alkyl of not more than 8 carbon atoms, have antibacterial properties, and some are comparable or superior to 1 i ncomyci n and can be used for the same purposes as lincomycin. The other analogues and isomers, including the compounds of formula IA and IB, have similar antibacterial properties but to a lesser degree and can be used for the same purposes as 1 i ncomyci n where larger amounts are not obj ectionabl e.

The following examples are illustrative of the process and products of the present invention but are not to be construed as limiting. The parts and percentages are by weight and the solvent ratios are by volume unless otherwise specified.

EXAMPLE 1 - 7-Deoxyl i neomycin A mixture of 10 g. of 7-chloro-7-deoxy1 i ncomyci n, 300 ml. of water, and 100 g. of zinc dust was stirred and heated at reflux for 15 hrs. The reaction mixture was cooled, filtered, and the filtrate freeze-dried to yield 8.0 g. of a white solid which was purified via chromatography over 1 kg. of silica gel as follows: Fraction ml_. MeOH:CHCl3 Forerun 1200 1.6 1-11 66 each discard 12-24 66 each P.A. 27¾6l/2 Fractions 12-2**· were combined and evaporated to give 1·5 &· of 7-deoxyllncom cJiin as a white solid.

Anal. Calcd. for CigH^NgO^St 55.351 H, 8.78| N, 7.17» S, 8.21.

In the following here is described the preparation of a number of 7-halo-7-deoxylincomycin derivatives* which are all novel compounds. All of these were subjected to a dehydrohalogenation process as set out in Example 1, and without any problems this process resulted in the corresponding 7-Heoxylincomycin derivatives! which were obtained in good yields.

PREPARATION L 7-Chloro-7-deoxylincomycin ^e h l 7-chloro-6,7»8-trideoxy-6-(trans-l~methyl-¾-propyl-L-2-.pyrrolidine-. carbo amido)-1·thio-I^threo-q-D-galacto-oetopyranoside· P.A.27*61/2 Α· The free base.

A suspension of 221.0 g. (0. 5 mole) of llncomycln hydrochloride In 5 1 · of -arbon tetrachloride was stirred well at 25° C. under nitrogen. 900 ml. of thlonyl chloride was added all at once and stirring continued for 2 hr. During this period the solid dissolved and a clear solution was obtained.

The reaction mixture was heated at reflux for 2 hr., the heat source then removed and nitrogen bubbled into the amber o solution until the pot temperature dropped to 25 C. About 4 1. of liquid was removed via vacuum di sti 1 lation at a pot o temperature of less than 5 C. The yellow solid which precipitated during this disti 1 lation was collected and dried. This o solid was dissolved In about 500 ml. of methanol, cooled at 25 C, made basic (pH IX). with dilute aqueous sodium hydroxide solution (2N), diluted to about 1200ml, with water and ex-tracted wel 1 wi th ether. The ether extracts were combined, washed with a small amount of water, dried over anhydrous magnes i urn sul fate and filtered. On evaporation of an aliquot of the combined ether extract 7-chloro-7-deoxyl 1 ncomyci n free base is obtained as a yellow amorphous solid, B. The hydrochloride Addition of hydrogen chloride gas to the filtrate of Part A resulted in the precipitation of 7-chloro-7-deoxy-lincomycin hydrochloride which was separated and recrys+al-lized from ethanol and ethyl acetate. A 52# yield of white, crystal ine 7-chl oro-7-deoxyl i neomycin hydrochloride solvated with approximately one molecular proportion of water was obtained, Anal . Calcd. for CieHaaCINgOsS' HC1 · H20: C, 45.18; H, 7.57; CI, 14.82; N, 5.86; S, 6.70; H20, 5.77.

Found: C, 44.70; H, 7.65; CI, 14,27; N, 5.78; S, 6.45j H20, 5.85.

H20 o . ~ [a] D + 150 (C-0.9858 g,/l00 ml,).

Activity: About 4 to 8 times li eomycin.

Antibacterial spectrum: Same as li neomycin.

PREPARATION 2 7-Chl oro-7-deoxyl i ncomyci n free base.

The process of Preparation 1, Part A, was repeated except that methylene chloride was used in place of ether as the extracting solvent and the combined extracts were filtered and evaporated to dryness. There was obtained a 64# yield of 7-chloro-7-deoxyl i neomycin free base as a yellow amorphous solid. 15 grams of this amorphous solid was adsorbed on JO grams of silica gel in methylene chloride, dried, and sifted onto a 3 inch diameter column of 1500 grams of silica gel. The column was then eluted with a mixture of methanol and chloroform in the proportions of 1 to 19, respectively, in 200 ml. portions after a 2 liter forerun. Fractions 26, 27» and 28 were pooled and evaporated to dryness yielding 1.04 grams of essentially pure 7-chloro-7-deoxy 1 i neomycin free base as an amorphous solid having an antibacterial spectrum of the same scope and magnitude as the 7-chloro-7-deoxyl i ncomyci n hydrochloride of Preparation 1.

PREPARATION 3 7-Halo-7-deoxyl i ncomyci n C [ethyl 7-chloro-6,7,8-trideoxy-6- (trans-l-methyl -4- propyl -L-2-pyrroiidi necarboxamido)-l-t h ί o- L -threo- a- D -galacto- octopy ra nos i de ] XX A-l. Chlorine substitution A suspension of 1 gram of l incomycin C hydrochloride in 25 ml. of carbon tetrachloride and .5 ml. of thionyl o chloride was stirred under nitrogen for 2 hours at 25 C.

A clear solution was obtained in about 15 minutes. The reaction mixture was then heated at reflux for 2 hours and evaporated to dryness under vacuum to give a yellow solid o which was dried under vacuum at 0 C. for l8 hours. The product was then dissolved in about 15 ml. of warm ethanol, made basic with sodium hydroxide as in Preparation 1 and diluted to 500 ml. with water. The water solution was extracted 5 times with 100 ml. portions of ether. The ether extracts were pooled, dried over magnesium sulfate, filtered, saturated with hydrogen chloride gas, and evaporated to a brown residue which on being recrystal 1 i zed two times from a mixture of ethanol and ethyl acetate (dissolved in a minimum amount of ethanol and ethyl acetate added to turbidity) yielded 200 mg. of 7-chl oro-7-deoxyl i ncomyc i n C hydrochloride as white crystals which had the same activity and spectrum as the 7-chl oro-7-deoxyl i ncomyci n hydrochloride of Preparation 1.

A-2. Halogen substitution A solution of Rydon reagent was prepared by stirring a dry solution of 52.6 g. (0.2 M) of triphenyl phosphi ne and o 800 ml. of acetonitrile at 50 under nitrogen and 10 ml. (0.19 ) of bromine added over a 20-min. period. After stirring for lO min. more, 8.2 g. of 1 i ncomyci n was added o and the reaction stirred at 50 for l8 hr. A white solid was then present. The reaction was filtered and the sol id discarded. Methanol (100 ml.) was added to the filtrate and the solvents then evaporated under vacuum. The viscous residue was dissolved in 100 ml. methanol, diluted with l800 ml. of water and extracted six times with 200 ml. portions of ether. The ether extracts were discarded, the aqueous phase made basic (pH 11 ) with aqueous KOH and then extracted four times with 200 ml. portions of methylene chloride. The extracts were dried and evaporated, leaving 11 g. of a yellow sol id which was chroma tog raphed over 1 kg. of silica gel, using methanol : chl oroform 1:9 (v/v) as the solvent system.

After a forerun of 1200 ml., 22 fractions of 56 ml . were collected. The last six (fractions 17-22) were pooled and evaporated to dryness yielding 2.8 g. of 7-bromo-7-deoxyl i neomycin. This was converted to the hydrobromide by dissolving in water, adding HBr to pH 1, filtering, and lyophilizing the filtrate. The hydrobromide had an Q + n4° (c, 0.9314, H20) and the following analysis: Calcd. for C1eH34Br2N205S: C, 39.28; H, 6.23; N, 5.09; S, 5.83; Br, 29.04.

Found: C, 39.64; H, 6.19; N, 5.07; S, 6.04; Br, 28.59.

In place of bromine, there can be substituted other halogens. Chlorine, for example, yields 7-chloro-7-deoxy-1 i ricomyci n whi ch is identical with the product obtained by chlorinating 1 incomycin with thionyl chloride. fn place of tri phenyl phosphi ne there can be substituted trlphenyl phosphi te. Also in that case a methyl halide can be used in place of the halogen. In place of the 1 incomycin, there can be substituted other lincomycins and analogs thereof. Thus, when 1 i ncomyci n C is substituted for 1 incomycin, 7- romo-7-deoxyl i ncomyci n C is obtained. y B. Preparation of Li neomycin C Li neomycin C is obtained by reacting I i ncomyci n wi th ethanethiol (ethyl mercaptan) to form a diethyl dithioacetal and heating the reaction mixture in the presence of p-toluene- sulfonic acid or heating to fusion. The following procedure is i 1 lust rat ive.

B-l. 6,8 Dideoxy-6- (trans-1 -methyl - -propyl -L-2-pyrrol i di ne- ca rboxam 1 do) -D-erythro-D-ga 1 acto-a 1 dehydo-octose diethyl dithioacetal .

In a l-liter, 3-necked flask were placed concentrated hydrochloric acid (150 cc. ) and ethanethiol (50 cc, previously o cooled to 0 ), followed by I i neomycin hydrochloride (15.0 gm. ). After stirring magnetically at room temperature for 5 hours, the reaction mixture was diluted with an equal volume of ice-water, and the solution extracted thoroughly with Skellysolve B (technical hexane), these extracts being discarded.

The majority of the acid was neutralized by the careful addition of solid potassium hydroxide (100 gm.), keeping the temperature of the wel l-stl rred reaction mixture be- o tween 20 and 30 C. by cool i ng 1 n acetone-Dry Ice-. Solid potassium chloride was removed by filtration, and the solid washed well with chloroform. Additional chloroform was added to the filtrate (ca. 150 cc. ) and the mixture, stl rred magnet- ically, was adjusted to pH 10 by the addition of aqueous sodium hydroxide (2 N). The chloroform layer was separated, the aqueous layer extracted thoroughly with chloroform, the combined extracts washed twice with water and dried over o anhydrous sodium sulfate. Removal of the solvent at 30 C. in vacuo gave a semi-solid residue, which on being crystallized from acetone, gave 5. l gm. of 6,8-di deoxy-6- (trans-1 -methyl -4- propyl -L-2-pyrrol id i necarboxami do) -D-erythro-D-gal acto-al dehydo-octose diethyl dithioacetal as colorless o flattened needles, m.p. 150-132 . Concentration of the mother o liquors gave additional material (1.50 gm. ), m.p. 129-151 . (Total yield, 6.91 gm., 42.4*).

Anal ysi s: Calculated for C21H 2N20eS2 : C, 52.25; H, 8.77; N, 5.8l; S, 13.29*.

Found: C, 52.38; H, 8.71; N, 5.93; S, 13.46*.

B-2. Cyclization to Lincomycin C. (a) One part each of the diethyl dithioacetal of Part B-1 and p-tol uenesul foni c acid monohydrate were refluxed in 25 parts of acetonitrile until substantial antibacterial activity was obtained. The react ion mixture was cooled and evaporated to dryness and chromatographed on sil ica gel using a solvent mixture ethyl acetate, acetone and water in the ratio of 8:5:1* respectively. Fractions 102 through 131 showed antibacterial activity. Of these fractions, 105 through 125 were pooled, evaporated to dryness, and crystallized from acetone acidified with hydrochloric acid and re-crystallized by dissolving in water and adding acetone to o give crystals of l incomycin C hydrochloride, m.p. 149-153 . (b) The diethyl dithioacetal of Part B-1 was heated r ° to 260 for about 3 minutes and the odor of ethyl mercaptan was noted. The product on being chromatographed as in Part B-2(a) yielded l incomycin C.

C. Preparation of Lincomycin C by fermentation. o The l incomycin C hydrochloride, m.p. 151-157 C. can also be prepared by the procedure described in Netherlands Patent 6,503,829 (Derwent l8,540) and A. D. Argoudelis et al ., Biochemistry 4, 70 -9 (1965).

D. Alternative method for preparation of Lincomycin C.

Lincomycin hydrochloride (8.85 g. - 0.02 moles) was o dissolved in 20 ml. of water, cooled at 0 and stirred while adding bromine (3.52 g. - 0.022 moles) dropwise over a 1 minute period. Ethanethiol (25 ml.) was added and the mix- o ture stirred at 25 for 2 hours. The clear, colorless, 2-phase system (ethanethiol is relatively insoluble in water) was cooled in an ice bath and hydrogen chloride gas bubbled in for about 5 minutes. The lower, aqueous phase turned red. The reaction mixture was then extracted 3 times with 100 ml. portions of Skellysolve B and aqueous sodium hydroxide solution added to bring the aqueous phase to pH 11. The basic phase was extracted well with chloroform. The chloroform extracts were washed with saturated sodium chloride solution, dried, and evaporated under vacuum to yield 6.2 g. of a white, sol id. 4.8 g. of this sol id was chromatographed over 800 g. of silica gel by the procedure of Preparation 2 using methanol -chloroform (1:7* respectively) as the solvent system. After 800 ml. of forerun, 80 fractions of 25 ml. each were col 1 ected. Fractions 40-58 were combined and evaporated to dryness and the residual sol id recrystal 1 i zed from acetone to yield 0.5 g. of material identical with the diethyl dithioacetal of Part B-l. Fractions 65-75 were combined, evaporated to dryness, and dissolved in a mixture of 5 ml . methanol and 00 ml. diethyl ether. Hydrogen chloride gas added and the white sol id which precipitated was collected. On being recrys ta 11 i zed from aqueous acetone, 0.5 g. of l incomycin C hydrochloride, identical with that of Part C, was obtained.

E . Other alkyl 7-halo-6,7,8-tr?deoxy-6- (trans-l-methyl - - propy 1 -L-2-pyrrol idi necarboxam? do)-l- thi o-L- threo-a-D- gal acto-octopyranos i des.

By substituting the ethanethiol in Part B-l and Part D of this preparation, with or without heating, by other alkyl mercaptans, for example, propyl, butyl, pentyl , hexyl, heptyl, octyl, nonyl, decyl, undecyl , dodecyl , tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl , octadecyl , nonadecyl, and eicosyl mercaptans and the isomeric forms thereof; by cyclo-alkyl mercaptans, for example, cyclopropyl, cyclobutyl, cyclo-pentyl, cyclohexyl, cycloheptyl, cyclooctyl, 2-methyl cyclo-pentyl, 2, 3-dimethyl cyclobutyl , 2-methyl cycl obutyl and 3-cyclo-pentylpropyl mercaptans; or by aralkyl mercaptans, for example, benzyl, phenethyl, 3-phenyl propyl , and 1-naphthylmethyl mercaptans, the corresponding alkyl, cycloalkyl, and aralkyl 6,8-dideoxy-6- (trans-l-methyl -4-propyl -L-2-pyrrol idi necarboxamido)-1- th io-D-ery thro-g-D- gal acto-octopyranos i des are obtained, which on treatment by the procedures of Preparation 1 and Part A are converted to the corresponding alkyl, cycloalkyl, and aralkyl 6,7,8-tri deoxy- and 7-halo-6,7,8-trideoxy-6- (t rans-l-methyl - -propyl -L- 2- pyrrol idi necarboxam i do) -1-th io-L-threo-g-D- gal acto-octopyranos i des. The compounds thus obtained (both the 6,8-dideoxy, the 6,7,8- trideoxy, and the 7-halo-6,7,8-trideoxy compounds) wherein alkyl is propyl, butyl, pentyl, and hexyl (obtained respectively when propyl, butyl, pentyl, and hexyl mercaptans are used) are effective antibacterials, having the same spectrum as lincomycin.

PREPARATION 4 Methyl 7-chloro-6,7,8-tr ideoxy-6- (trans -1 -a 1 kyl - -butyl -L-2-pyrrol idi necarboxamido)-l- thio-L-threo-a-D-galacto-octopyran-os i des.

R3 = methyl or ethyl.

Part A.

A suspension of ll6 mg. of methyl 6, 8-d ideoxy-6- (trans-1 -ethyl - -butyl -L-2-pyrrol idi neca rboxarni do) -l-thio-D-ery throes- D-ga lac to-octopyranoside free base in 2 ml. carbon tetra- o chloride and 0.7 ml . of thionyl chloride was stirred at 25 C. until a clear solution was obtained (about 15 minutes) and o allowed to stand at 25 C. for 2 hours. The reaction mixture was then heated at reflux for 2 hours and then evaporated to dryness under vacuum to a yellow sol id which was worked up as described in Example 3. The product, however, did not crystallize so the crystal 1 i zing solvents were evaporated to yield 17 mg. of methyl 7-chloro-6,7j8-trideoxy-6- (trans-l-ethyl - -butyl · L-2-pyrrol ldi necarboxamido)-l-thio-L-threo-a-D-galacto-octo-pyranoside hydrochloride as1 a tan amorphous solid.

On substituting the ci s epimer, there is obtained methyl- 7-chloro-6,7>8-trideoxy-6- (cls-l-ethyl - -butyl -L-2-pyrrol idi ne- ca rboxami do) -1-th i o-L- threo-a-D-gal acto-octopyranos i de hydrochloride having the same antibacterial spectrum.

On substituting the 1-methyl analogs, methyl 7-chloro-6,7j8-tri deoxy-6- (c i s- and trans -1-methyl - -butyl-L-2-pyrrol i-di necarboxami do)-l- thi o-L- threo-a-D-gal acto-octopyranos ides hydrochlorides are obtained.

The ci s and trans epimers used as starting materials in the above example were prepared as follows: Part B. - -Butyl idene-l-carbobenzoxy-L-prol i ne and the cycl ohexyl ami ne salt thereof.

Sodium hydride (19 g. ) as a 3$ suspension in mineral oil was warmed with 350 ml. of dimethyl sul oxide at a temper- o ature of 70-75 C. until the reaction was complete (about 30 o minutes). After cooling to 32 C, l6.2 g. of butyl triphenyl phosphonium bromide was added, and the resulting reaction mixture was stirred for 1 hour to insure complete reaction. A solution of 26 g. of -keto-l-carbobenzoxy-L-prol i ne in 100 ml. of dimethyl sulfoxide was added, and the resulting mixture o was heated at 70 C. for 3 hours. The reaction mixture was o cooled to 25 C. and 1 liter of 2.5$ aqueous potassium bicarbonate added. This mixture was washed twice with 700 ml. portions of ether and the ether was discarded after back extracting with 150 ml. of 2. $ aqueous potassium bicarbonate. The bicarbonate solutions were combined and acidified with N hydrochloric acid. The acidified mixture was extracted with four 500-ml . portions of ether. The combined ether extracts were washed successively with 250 ml. of water, three 250-ml. portions of saturated aqueous sodium bisulfite, and 250 ml. of water, and dried over anhydrous sod i urn su 1 fate. Evaporation of the solvent under vacuum gave 24 g. of an oily residue which was -butyl idene-l-carbobenzoxy-L-prol i ne.

This residue was dissolved in 31 ml. of acetonitrile and treated with l8 ml. of di cycl ohexylami. ne and refrigerated.

The crystals were collected, washed with acetonitrile and dried in vacuo giving 21 g. (46.8$) of the crystalline dicyclo- o hexylamine salt melting at 136-140 C. After two recrystal-lizations from acetonitrile, an analytical sample was obtained o o which melted at l42-l44 C. and had a rotation of [ ]Q - 4 Anal. Calcd. for 02ΘΗ44Ν204: C, 71.86; H, 9.15; N, 5.78.

Found: C, 71.69? H, 9.30; N, .7^· Ten grams of the dicyclohexylamine salt of 4-butyl i dene-1-carbobenzoxy-L-prol i ne was shaken with ether and excess $ aqueous potassium hydroxide until no solid remained. The layers were separated and each one was backwashed. The aqueous alkaline layer was combined with the backwash from the ether layer and acidified with 4N hydrochloric acid. The mixture was repeatedly extracted with ether and the ether extracts were combined, dried over sodium sulfate, and evaporated in vacuo to give 6.3 g. (9 56) of 4-butyl idene-l-carbobenzoxy-L-prol i ne as an oil.

Part C. - 4-Butyl -1-carbobenzoxy-L-prol i ne.

The oil from Part B was hydrogenated in 200 ml. of methanol over 2.1 g. of 10$ platinum on Dowex-1 catalyst under 40 lbs. hydrogen pressure. The catalyst was removed by filtration and the filtrate evaporated to yield 6.3 g. of 4-butyl -1-carbobenzoxy-L-prol i ne as an oil* The product contained about 2 parts ci s-4-butyl -1-carbobenzoxy-L-prol i ne to each part of trans-4-butyl -1-carbobenzoxy-L-prol i ne.

If desired, the hydrogenation of the 4-ylidene group can be postponed to any later step, even to the final step, By substituting the butyl triphenylphosphonium bromide of Part B by other substituted triphenylphosphonium bromides where the substituent is methyl, ethyl, propyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tri-decyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octa-decyl, nonadecyl, and eicosyl and the isomeric forms thereof; cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 2-cycl opropyl ethyl , and 3-cyclopentylpropyl ; benzyl, phenethyl, 3-phenyl propyl , and 1-naphthylmethyl , the correspond! ng 4-al kyl idene-, 4-cycl oal kyl ί dene-, and 4-aral kyl idene-l-carbobenzoxy-L-prol i nes and the corresponding 4-alkyl-, 4-cycloal kyl -, and 4-aral kyl -1-carbobenzoxy-L-prolines are obtained. For example, when the butyl triphenyl -phosphonium bromide is substituted by ethyl-, propyl-, i so-butyl-* pentyl-, and hexyltriphenylphosphonium bromides there are obtained 4-ethyl I dene-l-carbobenzoxy-L-prol i ne, 4-propyl- idene-l-carbobenzoxy-L-prol Ine, 4-isobutyl ldene-1-carbobenzoxy L-proline, 4-pentyl idene-l-carbobenzoxy-L-prol I ne, and 4-hexyl ldene-1-carbobenzoxy-L-prol i ne, and ci s and trans 4-ethyl -1-carbobenzoxy-L-prol I ne, 4-propyl -l-carbobenzoxy-L-proline, 4- i sobutyl -1-carbobenzoxy-L-prol I ne, 4-pentyl -1-carbobenzoxy-L-prol ine, and 4-hexyl -1-carbobenzoxy-L-prol I ne. Part D. - Methyl 6-ami no-6,8-di deoxy-l-thlo-D-erythro- g-D-galacto-octopyranoslde ( -MTL).

A solution of 40 g. of li neomycin free base (U. S.

Patent 3,086,912) In 20 ml. of hydrazine hydrate (98-10056) was refluxed for 21 hours; excess hydrazine hydrate was then removed i n vacuo under nitrogen at steam bath temperature, leaving a residue. The residue, a pasty mass of crystals, was cooled, acetonitrile was added, and the mixture was stirred until the crystals were suspended. The crystals were collected on a filter, washed with acetonitrile and with ether. The yield of white, crystalline a-MTL free base after drying i n vacuo at room temperature was 21 g. (84£). Recrystal 1 ization was accomplished by dissolving a-MTL free base in hot dimethyl formami de and adding an equal volume of ethylene glycol dimethyl ether.

Methyl 6-ami no-6,8-d ?deoxy-l-th? o-D-erythro-a-D-gal acto- _ o octopyranosi de free base has a melting point of 225-228 C, O an optical rotation of [O]Q + 276 (c = .768, water) and a pKa' of 7.45.

Anal. Calcd. for C9HT9N05S: C, 42.7; H, 7.56; N, 5.53; S, 12.66.

Found: C, 42.6; H, 7,49; N, 5.75; S, 12.38.

By substituting Vancomycin by other alkyl or by cyclo-alkyl or aralkyl 6,8-dideoxy-6- (trans-l-methyl -4-propyl -L-2-pyrrol idi necarboxam?do)-l-thio-D-erythro-a-D-galacto-octopy-ranosides where alkyl is ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl , nonadecyl, and eicosyl and the isomeric forms thereof; cyclo-alkyl is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 2-methyl cycl opentyl , 2,3-dimethyl -cyclobutyl, 2-methyl cycl obutyl , and 3-cycl opentyl propyl ; and aralkyl is benzyl, phenethyl, 3-phenyl propyl , and 1-naphthyl-methyl, the corresponding alkyl, cycloalkyl, and aralkyl 6-ami no-6,8-dideoxy-l-thio-D-erythro-g-D-galacto-octopyranos?des are obtained. For example, by substituting the li neomycin by ethyl-, propyl-, butyl-, pentyl-, and hexyl 6,8-dideoxy-6* (trans-l-methyl -4-propyl -L-2-pyrrol idi necarboxamido)-l-thio-D-erythro-g-D-galacto-octopyranosi des, ethyl 6-ami no-6,8- di deoxy-1- thi o-D-erythro-a-D-gal acto-octopyranos Ide, propyl 6 -ami no-6, 8-d ? deoxy-1- thi o-D-erythro-a-D -gal acto-octopyrano-side, butyl 6 -am? no-6, 8-d i deoxy-1 -t hi o-D-erythro-a-D-galacto-octopyranos i de, pentyl 6 -ami no-6, 8-d i deoxy-1- thi o-D-erythro-a-D-gal acto-octopyranos 1 de, and hexyl 6-ami no-6,8-di deoxy-1-t hi o-D-erythro-a-D -gal acto-octopyranos ide are obtained.

If desired, the procedure of Part A can be applied to the compounds of Part D using the hydrochloride or other salt of a strong acid and the resulting 7-halo compounds processed by the ensuing steps of this example to the final products of the preparation. Also, if desired, the resulting 7-halo compounds can be deshal ogenated by the procedure of Example 1 and the resulting 7-deoxy compounds processed by the ensuing steps of this preparation.

Part E. - Methyl 6,8-d?deoxy-6- (l-carbobenzoxy- - butyl - L -2 -pyrrol i d i necarboxami do)-l- th?o-D-erythro- g-D-galacto-octopyranos ide free base.

To a solution of 6.3 g. of -butyl -1-carbobenzoxy-L-proline (the oil from Part B) in 175 ml. of distilled aceton- o itrile cooled to 0 there was added 3. 6 ml. of tri ethyl ami ne followed by 3.3^ ml. of isobutyl chloroformate. The mixture o o was sti rred at 0 C. (+ 3 ) for 15 mi n. A solution .of 6.2 g. of a-MTL free base from Part C in 85 ml. of water was o added, and the reaction mixture was stirred at 0 C. for 0.5 o and at 25 C. for 1 hr. The reaction product was then filtered and dried yielding 4.57 g. (37.70 of methyl 6,8-dideoxy-6- (l-carbobenzoxy-4-butyl -L -2- pyrrol id i necarboxami do)-l- thio-D-erythro-g-D-gal acto-octopyranos ? de free base. The mother liquor was concentrated under vacuum and an additional 4.25 g. (35. #) of product recovered. Recrystal 1 i zat i on from acetonitrile produced crystals of methyl 6,8-d ί deoxy-6- (l-carbobenzoxy-4-butyl -L-2-pyrrol idi necarboxamido)-l>»thio-D-erythro-g-D- gal acto-octopyranos i de free base melting at 194- o 196 C. A second recrystal 1 ization f rom acetoni tri 1 e afforded o o an analytical sample, m.p. 195.5-200 C, [a]Q + ill (c, Ο.98, MeOH).

Anal. Calcd. for C2e o 20eS: C, 57.75; H, 7Λ6-, N, 5.13; S, 5.93.

Found: C, 57.58; H, 7.l6; N, 5.50; S, 6.07.

Part F. - Methyl 6, 8-dideoxy-6- (4-butyl -L-2-pyrrol idi necarboxami do)-l- thi o-D-erythro-g-D-gal acto- octopyranos i de hydrochloride.

A solution of 7.8 g, of methyl 6,8-dideoxy-6- (l-carbo-benzoxy-4-butyl -L-2-pyrrol id? necarboxami do) -1- thi o-D-erythro-g-D-galacto-octopyranosi de free base from Part E in 200 ml. of methanol was shaken over 2 g. of 10$ palladium on carbon under 40 lbs. of hydrogen pressure for 17 hours. The catalyst was removed by filtration and the solution concentrated under vacuum. The residue was dissolved in a mixture of 20 ml. of acetone and 20 ml. of water and acidified with 6N hydrochloric acid. Dilution with 4 volumes of acetone precipitated methyl 6,8-di deoxy-6- (4-butyl -L-2-pyrrol i di ne-carboxami do) -1- thio-D-ery thro-g-D-gal acto-octopyranos i de hydrochloride which was collected by filtration and dried. o The crystals, dried at 55 C. under vacuum, weighed 4.7 g. and melted at 188-194° C. The analytical sample obtained by recrystal 1 ization from acetone melted at 197-199° C. and gave [a]2Q5 + 150° (water, c, Ο.89).

Anal, Calcd. for Cter^NaOeS. HC1 : C, 48.80; H, 7.96; N, 6.32; S, 7.24.

Found: (corrected for .54$ water) C, 48.58; H, 8.19; N, 6.04; S, 7.36.

This material possesses 8$ of the antibacterial activity of 1 i ncomyci n . by S_. lutea assay.

By substituting the a-MTL by other alkyl or by cycloalkyl or aralkyl 6-ami no-6,8-d ideoxy-1- thio-D-ery thro- a- D-gal acto-octanopyranos ides wherein alkyl is ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tri decyl, tetradecyl, pentadecyl, hexadecyl, hepta-decyl, octadecyl, nonadecyl, and ei cosy 1 and the isomeric forms thereof; cycloalkyl is cyclopropyl, cyclobutyl, cyclo-pentyl, cyclohexyl, cycloheptyl, cyclooctyl, 2-methyl cyclo-pentyl, 2.,3-dimethyl cyclobutyl , 2-methyl cyclobutyl , and 3-cyclopentyl propyl ; and aralkyl is benzyl, phenethyl, 3-phenyl-propyl, and 1-naphthylmethyl , the corresponding alkyl, cycloalkyl, and aralkyl 6,8-di deoxy-6- (l-carbobenzoxy-4-butyl -L-2-pyrrol i dinecarboxami do)-l-th?o-D-erythro-a-D-galacto-octo-pyranosides and alkyl, cycloalkyl, and aralkyl 6,8-dideoxy-6- (4-butyl -L-2-pyrrol i d i necarboxam? do) -1- thi o-D-erythro- -D- ga I acto-octopyranos ides are obtained. For example, by substituting the a-MTL by ethyl, propyl, butyl, pentyl, and hexyl 6-am? no-6,8-d ideoxy-1 -thio-D-erythro-ct-D- gal acto-octo-pyranoside, there are obtained ethyl 6,8-di deoxy-6- (l-carbo-benzoxy-4-butyI -L-2-pyrrol id? necarboxami do)-D-erythro- -D-gal acto-octopyranos i de, propyl 6,8-dideoxy-6- (l-carbobenzoxy-4-butyl -L 2-pyrrol idi necarboxami do) -1-t i o-D-erythro- a-D~ ga1 acto-octopyranos i de, butyl 6,8-di deoxy-6- (l-carbobenzoxy-4-butyl-L-2-pyrrol idi necarboxami do)-l- thi o-D-erythro-a-D-galacto-octopyranos ide, pentyl 6,8-dideoxy-6- (1-carbobenzoxy-4-butyl -L-2-pyrrol idi necarboxami do)-l- thi o-D-erythro-a-D-galacto-octopyranosi de, hexyl 6,8-dideoxy-6- (l-carbobenzoxy-4-butyl -L-2-pyrrol i di necarboxami do )-l-thio-D-erythro-a-D-gal acto-octopyranos ide, ethyl 6,8-d ideoxy-6- (4-butyl -L-2-pyrrol idi necarboxami do) -1- thio-D-erythro-a-D-gal acto-octo-pyranoside, propyl 6,8-dideoxy-6- (4-butyl -L-2-pyrrol idi necarboxami do)-l-thio-D-erythro-g-D-galacto-octopyranosi de, butyl 6,8-dideoxy-6- (4-butyl -L-2-pyrrol idi necarboxami do) -1- thio-D-erythro-g-D-galacto octopyranos ide, pentyl 6,8-di deoxy-6- (4-butyl -L-2-pyrrol idinecarboxamido)-l-thio-D~ erythro-g-D-gal acto-octopyranos i de, hexyl 6,8-d ideoxy-6-(4-butyl -L-2-pyrrol idi necarboxami do) -1-th? o-D-erythro-a-D-gal acto-octopyranos ? de.

By substituting the 4-butyl -l-carbobenzoxy-L-prol i ne by other 4-al kyl -l-carbobenzoxy-L-prol i nes where the 4-alkyl ?s ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, and eicosyl and the isomeric forms thereof; by 4-cycl oal kyl -1-carboben-zoxy-L-prol i nes where 4-cycloal kyl is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 2-methyl-cyclopentyl, 2, 3-d imethyl cycl obutyl , 4-methyl cycl obutyl , and 3-cycl opentyl propyl ; and by 4-aral kyl -1-carbobenzoxy-L-pro- 1 i nes where 4-aral kyl Is benzyl, phenethyl, 3-phenyl propyl , and 1-naphthylmethyl , the corresponding alkyl, cycloalkyl, and aralkyl 6,8-d i deoxy-6- (l-carbobenzoxy-4-al kyl , 4-cyclo-al kyl , and 4-aral kyl -L-2-pyrrol idi necarboxami do)-l-thio-D-erythro-g-D-galacto-octopyranos i des, and the corresponding alkyl, cycloalkyl, and aral kyl -6- (4-al kyl , 4-cycl oal kyl , 4-aral kyl -L-2-pyrrol idi necarboxami do)-l-thi o-D-erythro- -D-ga 1 acto-octopy ranos ? des are obtained. For example, by substituting the 4-butyl -1-carbobenzoxy-L-prol i ne by 4-methyl-, 4--ethyl-, 4-propyl-, 4-pentyl-, and 4-hexyl -1-carbobenzoxy-L-prol i nes, there are obtained methyl, ethyl, propyl, butyl, pentyl, and hexyl 6,8-dideoxy-6- (l-carbobenzoxy-4-methyl -L-2-pyrrol i d i necarboxami do)-1-th io-D-erythro-a-D- gal acto-octo-pyranosides; methyl, ethyl, propyl, butyl, pentyl, and hexyl 6,8-dideoxy-6- (l-carbobenzoxy-4-ethyl -L-2-pyrrol idi necarboxami do) -1-th io-D-ejy_thro- -D-^gJ_a to-octopyranos i des ; methyl , ethyl, propyl, butyl, pentyl, and hexyl 6,8-d i deoxy-6- (l-carbobenzoxy-4-propyl -L-2-pyrrol idi necarboxami do)-l-thio-D-erythro-g-D-gal acto-octopyranos ides; methyl, ethyl, propyl, butyl, pentyl, and hexyl 6,8-d i deoxy-6- (l-carbobenzoxy-4-pentyl -L-2-pyrrol id i necarboxami do)-1-th? o-D-erythro-a-D-ga 1 acto-octopy ranos } des ; methyl, ethyl, propyl, butyl, pentyl, and hexyl 6,8-dideoxy-6- (l-carbobenzoxy-4- hexyl -L-2-pyrrol i -dl necarboxami do)-1-th i o-D-erythro-g-D -gal acto-octopyranos i des ; methyl, ethyl, propyl, butyl, pentyl, and hexyl 6,8-dideoxy-6- (4-methyl -L-2-pyrrol idi necarboxami do)-1-th t o- D-erythro-g-D-ga 1 acto-octopy ranos i des ; methyl, ethyl, propyl, butyl, pentyl, and hexyl 6,8-d ideoxy-6- ( -ethyl -L-2-pyrrolidi necarboxami do) -1- thio-D-ery thro- ct-D- gal acto-octopyranos i des ; methyl, ethyl, propyl, butyl, pentyl, and hexyl 6,8-d ideoxy-6- ( -propy I -L-2-py r rol i d i necarboxami do) -1-t hi o-D-erythro-a-D- gal acto-octo-pyranosides; methyl, ethyl, propyl, butyl, pentyl, and hexyl 6,8-d i deoxy-6- ( -pentyl -L-2-pyrrol id i necarboxami do) -1-th i o-D-erythro-a-D-gal acto-octopyranos i des; and methyl, ethyl, propyl, butyl, pentyl, and hexyl 6, 8-di deoxy-6- ( -hexyl -L-2-pyrrol ? di necarboxam f do)-l-thio-D-erythro-a-D-gal acto-octopy-ranosides.

If desired, the 1-carbobenzoxy compounds prepared according to Part E can be halogenated by the procedures of Part A, followed by deshalogenation by the procedure of Example 1 if desired, and the resulting 7-halo and 7-deoxy compounds processed by the ensuing steps of the preparation to remove the 1-carbobenzoxy group and to substitute the prol ine nitrogen to give 7-halo or 7-deoxy products.

Part G-l. - Methyl 6,8-d i deoxy-6- (l-methyl - -butyl -L-2- pyrrol id i necarboxami do) -1-th i o-D-erythro-a-D gal acto-octopy ranos ? de hydrochloride.

A solution of 2.0 g. of methyl 6, 8-di deoxy-6- ( -butyl -L -2- pyrrol i di necarboxam i do) -1-th i o-D-erythro-a-D-galacto-octopyranoside hydrochloride from Part F and 2.0 ml. of 37# formalin in 150 ml. of methanol was shaken over 500 mg. of 104 palladium on carbon under 40 lbs. of hydrogen pressure for 3.5 hrs. Removal of the catalyst by filtration and the solvent by distillation in vacuo yielded partially crystalline methyl 6,8-dideoxy-6- (l-methyl -4-butyl -L-2-pyrrol idi necarbox-am i do) -l-thio-D-erythro-a-D-gal acto-octopyranos ide hydrochlor-ide which by TLC (thin layer chromatography) on sil ica gel using a mixture of ethyl acetate, acetone, water (8:4:l) for el ut ion and Mn04 solution for detection consisted chiefly of two materials, the ci s and trans epimers of methyl 6,8-dideoxy-6- (l-methyl -4-butyl -L-2-pyrrol idinecarboxamido)-l-thio-D-erythro-g-D-gal acto-octopyranos ide hydrochloride in a ratio of about 3 to 2.

G-2. Separation of the ci s and t;rans forms by chromatography.

The methyl 6, 8-dideoxy-6- (l-methyl -4-butyl -L-2-pyrrol i -di necarboxam? do) -1-th io-D-erythro-a-D-gal acto-octopyranos ide hydrochloride from Part G-l were dissolved in a mixture of methanol and methylene chloride (l:l) and 1.5 ml. of triethyl-amine added. To this solution was added 7 g. of silica gel and the solvent evaporated under vacuum leaving the ant i bi ot i c deposited on the silica gel which was sifted on top of a chromatographic column of 200 grams of silica gel packed with a solvent mixture consisting of ethyl acetate, acetone, water in a ratio of 8:4:1. The column was developed by eluting with the same solvent and 20 ml. portions were collected.

Thin layer chromatography of each fraction showed that fractions 31-38, 310 mg., were essentially pure trans epimer, that fractions 49-74, 32 mg., were essentially pure ci s epimer, and that fractions 39-48 consisted of a mixture of epimers.

The latter could be further separated by repeated chromatography. Each epimer was dissolved in a few drops of dilute hydrochloric acid and the hydrochloride precipitated by addition of acetone.

In this manner, there was obtained 50 mg. of methyl 6,8-di-deoxy-6- (trans-l-methyl -4-butyl -L-2-pyrrol idi neca rboxam i do) - 1- th? o-D-erythro-a-D-galacto-octop ranos i de hydrochl or i de, o m.p. 155-137 > and about 150 mg. of methyl 6,8-dideoxy-6- (ci s-l-methyl -4-butyl -L-2-pyrrolidi neca rboxam i do)-l- thi o-D erythro-a-D-gal acto-octopyra osi de hydrochloride, softening o o at 105 C. with further melting at 175-185 C.

The trans epimer recrystal 1 i zed from the same solvent o melted at 139-141 C. and had the following analysis: Anal. Calcd. for ΰ1ΘΗ3βΝ20β5. HC1 : C, 9.93; H, 8.l6; N, 6.13; S, 7.02.

Found: (Corrected for 4.07# H20); C, 48.81; H, 8.54; N, 6.49; S, 6.67.

Similarly recrystal 1 ization of the ci s epimer gave a o o product softening at 108 C. and further at about 189 C. (solvated) with the following analysis: Anal. Found: (Corrected for 4.95$ water); C, 50.27; H, 9.00; N, 6.05; S, 6.65.

The trans epimer was about 2.2 times as active as li neomycin by . 1 utea assay, about 2 times as active by the broth dilution assay, and 2.5 times as active in mice infected with aureus.

The c? s epimer was about 1/2 to 1/3 as active as the trans epimer, being about equal to l incomycin.

H-l. - Methyl 6,8-dideoxy-6-"(l-ethyl -4-butyl -L-2-pyrrol -i d i neca rboxam ?do)-l-thio-0 -erythro- a- D -galacto- octopyranoside hydrochloride.

A mixture of 2,0 g. of methyl 6, 8-deoxy-6- (4-butyl -L-2-pyrrol idi necarboxamJdo)-l-thio-D-erythro-ct-D-ga1acto-octopyrano- i side hydrochloride from Part F, i.5 ml. of acetal dehyde, 150 mg. of 10$ palladium on carbon in 150 ml. of methanol was shaken under 3 lbs. of hydrogen pressure for 5· 5 hrs. The catalyst was removed by filtration to give a residue consisting chiefly of the c? s and t rans epimers of methyl 6,8-dideoxy-6- (1-ethyl -4-butyl -L-2-pyrrol idi necarboxami do) -1- thio-D-erythro-a-D-galacto-octopyranos i de hydrochloride.

H-2. - Separation of epimers.

As described in Part G-2, the mixture of epimers of Part H-l (2 g, ) was chroma tographed over 200 g. of silica gel using for el ut ion a sol vent system- of ethyl acetate, afcetone, water (8:4:1). Fractions 33-42 by TLC were pure t rans-ep ?mer and were combined, fractions 49-64 were essentially pure ci s-ep? -mer and were also combined. Fractions 43-48 were a mixture of the epimers which could be purified by rechromatography.

Each epimer was dissolved in a few drops of diltrte hydrochloric acid and the crystalline hydrochloride precipitated on dilution with a large volume of ether.

The crude trans epimer fraction of 415 mg. gave 340 mg. (15,4$) of crystal 1 i ne methyl 6,8-d? deoxy-6- (trans-1-ethyl -4-buty1 -L -2-pyrro1 i d i necarboxam ido)-l-th io^P-erythro- -D-galacto- o octopyranoside hyd ochloride, m.p. 144-151 C. Recrystal 1 iza- o tion from dilute acetone ra i sed the m.p. to 148-151 C.

The cis epimer fraction of 645 mg. afforded 300 mg. (l4.l£) of crystalline methyl 6,8-dideoxy-6- (cis-l-ethyl -4-butyl -L-2-pyrrol idi nec rboxami do)-1-thio-D-erythro-ct-D-gal acto- o octopyranoside hydrochloride, m.p, 135-139 C. Recrystal 1 iza- o tion from dilute acetone gave crystals, m.p. 134-138 C.

The trans epimer isomer showed about 1-1.2 times the activity of 1 i neomycin by the lu ea assay, 2-4 times the activity of I i ncomyci n aga i nst Gram positive organisms, and 8 times or more the activity of 1 i ncomyc i n aga i nst Gram- negative organisms. In mice against JS. aureus the t rans epimer was about 2 times as active as lincomycin. The ci s epimer was about 1/2 as active as the trans epimer.

Separation of the ci s and trans isomers is not a necessary step as the 7-chloro derivatives of the mixed epimers are useful per se. It is desi able, however, to keep the content of trans isomer high as this is the most active form. By carrying out the process with this in mind mixed epimeric products containing a ratio of trans and ci s epimers of 3:1 to 1:5 can readily be obtained. By substituting the formaldehyde and acetaldehyde of Parts G and H by other oxo compounds of the formula R4R5CO, for example, propi onal dehyde, acetone, butyral dehyde, isobutyl methyl ketone, benzal dehyde, phenylacetal dehyde, hydroci nnamal dehyde, acetophenone, pro-piophenone, butyrophenone, 3-methyl -4-phenyl -2-butanone, 2-methyl -5- phenyl -3-pentanone, 3-cyclopentanepropi onal dehyde, cyclohexaneacetal dehyde, cycl oheptanecarboxa 1 dehyde, 2,2-di-methyl cyclopropaneacetaldehyde, 2, 2-d imethyl cycl ©propyl methyl ketone, cyclopentyl methyl ketone, cyclobutyl methyl ketone, cyclobutanone, cycl hexanone, and 4-methyl cyclohexanone, and using the appropriate alkyl, cycloalkyl, or aralkyl 6,8-dideoxy-6- (4-al kyl , 4-cycloal kyl or 4-aral kyl -L-2-pyrrol i -di necarboxamido)-l-thi o-D-erythro-a-D-galacto-octopyranos?de, there are obtained the corresponding alkyl, cycloalkyl, and aralkyl 6,8-dideoxy-6- (l-R4R5CH-4-a1 kyl , 4-cycloal kyl and 4-aral kyl -L-2-pyrrol i di necarboxamido)-!- thi o-D-erythro-a-D-gal acto-octopyranos i des which on treatment with thionyl chloride by the procedure of Part A gives the corresponding alkyl, cycloalkyl, and aralkyl, 7-chloro-6,7,8-trideoxy-6- (l-R4R5CH-4-al kyl 4-cycl oal kyl and -a ra 1 ky 1 - L-2- py r rol idfne-carboxami do) -1-th io-L-th reo-a-D-gal acto-octopyranos ides where R4R5CH- is propyl, isopropyl, butyl, and 4-methyl -2-pentyl ; benzyl, phenethyl, 3-phenyl propyl , 1-phenyl ethyl , i-phenyl-propyl, 1-phenyl butyl , 2-methyl -4-phenyl -2-buty 1 , and 2-methyl -5-phenyl -5-pentyl ; 3-cycl opentyl propyl , 2-cycl ohexyl -ethyl, cycloheptylmethyl , 2- (2,2-dimethylcyclopropyl )-ethyl , 1- (2,2-dimethyl cyclopropyl )ethyl , 1-cyclopentyl ethyl , 1-cyclo-butylethyl, cyclobutyl, cyclohexyl, and 4-methyl cycl ohexyl . By using formaldehyde and acetaldehyde or other alkanals, for example, propi onal dehyde, huttyral dehyde, val eral dehyde, or caproaldehyde with an al kyl 6,8-d ideoxy-6- (4-al kyl -L-2-pyrrol i d i necarboxamido) -1- th o-D-erythro-a-D-galacto-octopy-ranoside where al kyl and 4-alkyl are methyl, ethyl, propyl, butyl, pentyl, or hexyl, preferred starting compounds of the formula wherein X is hydroxy; R, H t, and R3 are alkyl of not more than six carbon atoms, advantageously of not more than twelve carbon atoms in the aggregate; and the configuration is D-erythro, are obtained which on treatment with thionly chloride or Rydon reagent by the procedure of Part A give compounds according to Formula XXIX where X is halogen; and which on further treatment with zinc dust by the procedure of Example 1 gives compounds of Formula XXIX wherein X is hydrogen; R, H i, and R3 in both cases are alkyl as given above; and the configuration is believed to be L-threo. The 6,8-dideoxy, 6,7*8-trideoxy, and 7-halo-6,7»8-trideoxy compounds of formula XXIX as characterized above are antibacterial s and have the same spectrum of antibacterial activity as l i neomycin. Representative compounds of Formula XXIX wherein X is hydrogen but which are bel ieved to have the L-threo configuration are given in the following table: TABLE I COMPOUNDS OF FORMULTTYTSTWHE RE X IS HYDROGEN 4A methyl trans-ethyl methyl 4B methyl ci s-ethyl methyl 4C f7-deoxyl i ncomyci n) methyl trans-propyl methyl 4D 7-deoxyal lol ί ncomyci n) methyl ci s-propyl methyl 4E f7-deoxyl i ncomyci n E) methyl trans-propyl ethyl 4F [ 7-deoxya 11 ol i neomycin E) methyl ci s-propyl ethyl 4G ethyl trans-propyl methyl 4H ethyl ci s-propyl methyl 41 methyl trans-butyl methyl 4J methyl ci s-buty 1 methyl 4K methyl trans-propyl ethyl 4L methyl ci s-propyl ethyl 4M ethyl trans-propyl ethyl 4N ethyl ci s-propyl ethyl 40 methyl trans-butyl ethyl 4P methyl ci s-butyl ethyl 4Q methyl trans-pentyl methyl 4R methyl ci s-pentyl methyl 4S ethyl trans-butyl ethyl 4T ethyl ci s-butyl ethyl 4U methyl trans-pentyl ethyl 4V methyl ci s-pentyl ethyl 4W ethyl trans-pentyl methyl 4X ethyl ci s-pentyl methyl 4Y methyl trans-hexyl methyl 4Z methyl ci s-hexyl methyl 4AA butyl trans-propyl methyl 4AB butyl ci s-propyl methyl 4AC ethyl trans-pentyl ethyl 4 AD ethyl ci s-pentyl ethyl 4AE butyl trans-butyl ethyl TABLE I (Continued) R HRi Ra 4AF butyl cl s-butyl ethyl 4AG butyl trans-pentyl methyl 4AH butyl cis-pentyl methyl 4AI cycio- hexyl trans-propyl methyl 4AJ cyclo- hex l cis-propyl methyl 4A butyl trans-pentyl ethyl 4AL butyl ci s -pentyl ethyl 4AM pentyl trans-pentyl ethyl 4AN pentyl cis-pentyl ethyl Intermediates for the preparatioh of the above compounds correspond to the above table where (l) X is halogen; (2) R3 is hydrogen; (3) X Is halogen and R3 Is hydrogen; (4) X is halogen and R3 is carbobenzoxy; (5) X is halogen/ R3 is carbo-benzoxy, and HRt and H In the 4-position are replaced by the ylidene group, Rt ; (6) X is halogen, R3 is hydrogen, and HRX and H in the 4-position are replaced by the ylidene group, Rt; and (7) Is halogen, and HRj and H In the 4-position are replaced by the ylidene group, R*; and It being understood that the configuration is believed to be L-threo. The corresponding compounds where X is hydroxy having the D-erythro configuration are used as starting compounds.

When the 11 neomycin of Part A -2 of Preparation 3 is replaced by alkyl, cycloalkyl, or aralkyi 6-ami no-S^S-dideoxy-l-thlo-D-ej^thj^- -D-^aJjacto-octopyranos ide free base there are obtained compounds of the formula: XXX wherein X is hydrogen or halogen, R is ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tri decyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octa-decyl , nonadecyl , and eicosyl, or any of the isomeric forms thereof, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 2-methyl cycl opentyl , 2,5-dimethyl -cyclobutyl, 2-methy1cyclobutyl , and 3-cycl opentyl propyl ; benzyl, phenethyl, 5-phenyl propyl , and 1-naphthyl methyl which can be acylated by the procedures of Preparation 4 to form the corresponding 7-deoxy- and 7-chloro-7-deoxyl i ncomyci n analogs, for example, the compounds of Formula XX I X as given and illustrated above, as well as the intermediates therefor.

PREPARATION 5 Methyl 7-halo-6,7,8-trideoxy-6- (tjrans-1 -methyl -4-propyl -L-2-pyrrol id i necarboxaml do)-l- thio-D-erythro-a-D-galacto-octopy-ranoside (7-halo-7-deoxy-epi 1 i ncomyci n) .

A. 5*4-0- 1 sbpropyl idenel i neomycin.

A solution of 9.8 g. of 1 i ncomyci n i n 150 ml. of acetone is added to a solution of 9.8 g. of p-tol uenesul foni c acid monohydrate in 100 ml. of acetone with good stirring and avoidance of exposure to moisture. The mixture is stirred at ambient temperature for 1 hour, after which 100 ml. of anhydrous ether !s added and stirring is continued in an ice-bath for 0.5 hour. The mixture is filtered and the solid is dried in vacuo at 50° C. ; yield 15,35 g. (85.5*) of 3,4-0- i sopropyl i denel i ncomyci n p-tol uenesul fonate. An additional 1.15 g.; (7. 5?) can be recovered from the mother liquors by adding 350 ml. of anhydrous ether to the mother liquor from the previous filtering operation and chilling the solution for 1 hour. The 14.5 g. so obtained are suspended in 200 ml. of ether and shaken vigorously with 125 ml. of * potassium bicarbonate solution. The aqueous layer is back-extracted with two 100-ml. portions of ether. The ether extracts are washed with 50 ml. of saturated sodium chloride solution and then filtered through anhydrous sodium sulfate. The ether is evaporated under vacuum, leaving 7.9 g. (73.1*) of 3*4-0-i sopropyl i denel i ncomyci n which is dissolved in 25 ml, of ether, acetate and concentrated to about J.0 to 15 ml. The concentrate is allowed to stand at room temperature for several hours and then refrigerated overnight. The crystals are filtered from the solution and washed sparingly with cold ethyl acetate; yield 4.55 g. (42.2£) of 3,4-0- i sopropyl i dene- 1 i neomycin having a melting point of 126-128 Cf, and an optical rotation of [a] * 101-102 (c, 1, methylene chloride).

B. 7-Dehydro-3,4-0- i sopropyl i denel i ncomyci n.

To a solution of 6 g. (0.0135 moles) of i sopropyl idene-1 i ncomyci n : i n ,75 ml . of pyridine was added 12 g. (excess) o chromic oxide. The solution warms up about 20 C. After one hour the mixture was added to a solution containing 250 ml, each of ethyl ether and ethyl acetate. This was then filtered and evaporated to a syrup, 8.4 g. This syrup was distributed in a 500-transfer counter current distribution using the system, water.ethyl acetate: ethanol :cyclohexane (1:1:1:1). 7-Dehydro-3,4-0- i sopropyl i denel i ncomyci n was isolated as the peak fraction from tubes 330-380, = 2.45.

Anal. Calcd. for CaiHaeNaOeS: C, 56.72; H, 8.l6; N, 6.30; S, 7.21.

Found: C, 56.37; H, 7.62; N, 6.51; S, 6.84.

C. 3*4-0-1 sop ropy 1 idene-epi 1 i ncomyci n.

To 1.6 g. of Craig-pure 7-dehydro-3* 4-0- i sopropyl i dene- li neomycin in 75 nil. of methanol was added 400 mg. of sodium borohydride. After 1.5 hr. this solution was evaporated to dryness on a rotary evaporator. The residue was added to 25 ml. of water and extracted three times with 25 ml. each of methylene chloride. The extract was back-washed with 15 ml. of water, then dried over magnesium chloride and evaporated to dryness. The residue, 1.4 g,, was distributed in a 500-transfer counter current distribution using the solvent system, water:ethyl acetate: ethanol :cycl ohexane (1:1:1:1), and a single peak which fit the theoretical was observed at K = 1.05. The material in tubes 240 to 280 was isolated as a syrup.

Anal. Calcd. for C21H38N20eS: C, 56.47; H, 8.58; N, 6.27; S, 7.l8.

Found: C, 56.24; H, 8.54; N, 6.13; S, 7.01.

Thin layer chromatography (TLC) showed that this material consisted of two substances. One was 3*4-0- i sopropyl -i denel i ncomyci n; the other, 3, 4-0- i sopropyl idene-epi 1 incomyci n, moved slightly slower.

D. Epi 1 incomycin.

The syrup from Part C was stored at room temperature hrs. in a solution containing 60 ml. of 0.25 ,Ν hydrochloric o acid and 40 ml. of ethanol. It was then kept at 0 C. for 4 days. Following neutralization with sodium bicarbonate, it was evaporated to 25 ml., then extracted with chloroform. The extract was washed with a little water and dried over magnesium sulfate, then evaporated to a residue. Thin layer chromatography of the residue showed two substances, both of which were active against . 1 utea. The residue was chroma tog rap hed on a 14" x 3/4" Florlsll (a synthetic silicate of the type described In U. S. Patent 2,593*625) column which was eluted gradiently with solvent which varied continuously from 100% Skellysolve B (technical hexane) to 100% acetone. The total volume was 5000 ml. The two compounds were thus separated.

Fraction I: Tubes 53-65 ( 0 ml. cuts) Epi 1 i ncomyci n.

Assay 450 meg. /mi.

Anal. Calcd. for CtaH^NaOeS: C, 50.92; H, 8.55; N, 6.60; S, 7.56.

Found: C, 50.19; H, 7.91 N, 6.05; S, 6.42.

Fraction II: Tubes 73-104. Llncomycin Assay 950 meg. /mg.

E. 7- Chi oro-7-deoxyepi 1 i ncomyci η· HCl .

XXXII 7-Epl 11 neomycin hydrochloride (0.85 g.) was suspended in 17 ml. of carbon tetrachloride. Thionyl chloride (4,5 ml.) o was added and the reaction mixture stirred at 25 for 15 minutes. The solid dissolved to give a clear, colorless solution, The reaction was then heated at reflux for 2 hours, cooled to o , evaporated under vacuum to a solid yellow residue. This crude product was dissolved in 10 ml. of ethanol, made basic with 0.1N sodium hydroxide, diluted to 500 ml* with water and extracted four times with 50-ml . portions of chloroform. The combined chloroform extracts were back-washed two times with 20-ml. portions of saturated sodium chloride solution, filtered and evaporated under vacuum. The solid residue was slurried with 500 ml. of ether, filtered and hydrogen chloride gas passed into the filtrate. The solid precipi tate was collected, dissolved In 5 ml. of ethanol , and re-precipitated by the addition of ether.

The solid was collected and dried. A 20$ yield (270 mg. ) of 7-chloro-7-deoxyepl 11 neomycin assaying about 2 to 3 times the actl l ty of .11 neomycin was obtained. TLC on silica gel (MeOHtCHCla system, 1:6 v/v) showed one spot having an Rf of 0.44 as compared with an Rf of 0.52 for 7-chloro-7-deoxyl i neomycin.

Following the procedure of Preparation 1, Part A-2, 7-bromo-7-deoxy1 I eomycin is obtained.

By substituting 11 neomycin In this preparation by li neomycin analogs of Formula II wherein Z, R, Rt, Rg, R3 of the Ac group are as given and Illustrated above, the corresponding 7-hal 0-7- deoxyep.i 1 i neomycin analogs of the formula: XXXIII wherein X is halogen and Z, R, Rt, R2, and Ra of the Ac group are as given and Illustrated above, are obtained. All the compounds that have been described above, therefore, have their counterpart in the opposite configuration, that is, the configuration derived from the 7-epi form. If an inversion is effected by the substitution of the 7-hydroxy group by halogen, then the epi -compounds which have the L- threo configuration, are inverted to the D-erythro configuration. In any event both the D-erythro and the L- threo forms are obtained, the one when the normal lincomycins (D-erythro) are used, the other when ep i - 1 i ncomyc i ns (L-threo) are used. Both forms, however, yield the same 6,7,8-t ri deoxy compound on des ha 1 ogena t i on .

While the processes of the invention have been described with reference to the preparation of specific compounds, it is to be understood that the process is broadly applicable to compounds of Formula II wherein Ac and are radicals which are not reactive with thionyl chloride. Thus, in the broad process according to the invention, Ac can be hydrogen or any acyl that is non-reactive with thionyl chloride and R can be any alkyl, irrespective of the number of carbons, or any other radical, for example, aralkyl like benzyl, naphthylmethyl , and benzhydryl, or like hydrocarbon radicals that are non-reactive with thionyl chloride.

Claims (1)

1. 2138 HAVING NOW particularly described and the na ure of our said invention and in what manner the same is to be we declare that what we claim is A compound of the formula CH3 wherein R i s of not more than 20 carbon atoms and Ac hydrogen or the radical of a 4 ubs t i t uted r ro 1 carboxyl i c acid of the formula wherein Ri and are selected f rom the group consi sting of and alkyl and cycloalk l a 1 not more than 20 carbon eye 1 oa 1 ky 1 i dene and of f rom 3 to not more than 8 carbon a 1 ky 1 i not more than 12 carbon atoms Z i s a protective hydroca rbony 1 d i phenyl 1 methoxypheny 1 benzyl or trobenzyl and R3 is selected from the group cons isti ng of hydrogen and V A compound according to Claim 2 wherein R is and is and trans trans compounds as defined in Claim substantially as hereinbefore described and reference to any of the The process of making compounds of the formula wherein and Ac are as given the step of reacting a compound of the formula OH zinc dust at a temperature effective to remove halogen Ac acyl with zinc or where HX is a strong acid and R is compounds defined in Claim whenever obtained by a process as claimed in claim FOR APPLICANT insufficientOCRQuality