BE705364A - - Google Patents

Description

  

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  "Lincomycin derivatives, analogs and isomers and methods of preparation"
The present invention relates to compounds and processes for their preparation, and more particularly relates to 7-halo-7-deoxylincomycin and 7-halo-7-deoxyepilincomycin, analogs and isomers thereof. , as well as the processes by which these compounds and analogous compounds are prepared.



   The new compounds of the invention can be represented by the following structural formula

  <Desc / Clms Page number 2>

 
 EMI2.1
 in which R represents an alkyl of 20 carbon atoms at most, advantageously of 8 carbon atoms at most, a cycloalkyl of 3 to 8 carbon atoms at most, or an aralkyl of 12 carbon atoms at most, advantageously of 8 atoms at most carbon, and Ac is the acyl radical of a substituted L-2-pyrrolidinecarboxylic acid in position 4, having the formula:

     
 EMI2.2
 formulas in which R1 and R2 represent an alkylidene of at most 20 carbon atoms (in particular methylene), preferably of 8 carbon atoms at most, a cycloalkyldene of 3 to 8 carbon atoms at most, or an aralkylidene of 12 atom. of carbon at most, advantageously of 8 carbon atoms at most, and R3 represents hydrogen or HR2.



   Examples of alkyls having 20 carbon atoms at most (R, HR1 and HR2) are: methyl, ethyl, propyl, butyl, pentyl hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, and eicosyl and their isomeric forms. Examples of cycloalkyls are cyclopropyl, cy-
1

  <Desc / Clms Page number 3>

 
 EMI3.1
 dobutyl, cyclopentyl, cyclohexyl, oycloheptyl, cYcloOctYl, 2-methylcyolopentyl, 2,3-dim6thylcyclobutyl, 4-methylcyclobutyl, and 3-cyclopantylpropy.a.

   Examples of alkylidene, cycloalkµlidene and 4.! aralkylidene (R1 and R2) are methylene, ethylene, propylidene butylidene, pentylidene, hoxylidene, heptylidr.e, octylidene, nonylidene, decylidene, undecylidene, dodecylidene, tridecylidene, tetradecylidene, 3idelideneadaylidene, heptylidene, decylidene, undecylidene, dodecylidene, 3idelideadaylidene, octylidene, heptylidene, pencylidene, hepidecidadaylidene, hepidecaylidene, hepidecylidene, hepidecaylidene, octylidene, hepidadidene eicosylidene, and their forez my isomers, cyclopropylidene, cyclobutylidene, cyclopentylidene, cyclohexylidene, cycloheptylidènel cyclooctylidene, 2-cyclopropylethylidene, 3-cyclopentylpropylidene, benzylidene, 2-phenyl-phenyl-ethylidene and propylidene, naphthylidene, 3-phenyl-phenyl-3-ethylidene and 3-methylidene, 2-phenyl-phenyl-ethylidene and 3-methylidene, 2-phenyl-phenyl-ethylidene and 3-methylidene, 2-phenyl-phenyl-ethylidene, 2-phenyl-phenyl-ethylidene, 3-cyclopropyl-propylidene, 2-phenyl-phenyl-ethylidene-methylidene, 2-phenyl-phenyl-ethylidene, 3-phenyl-phenyl-ethylidene, 3-cyclo-propylidene, 3-cyclopentylpropylidene.



   The novel compounds of the invention (formula I), as well as other related compounds, can be prepared by replacing with chlorine the 7-hydroxy of a compound of the formula:
 EMI3.2
 
The replacement is advantageously carried out by mixing the starting compound of formula II with a Rydon reagent, and heating. In the process, Ac and R can be any radical which is not reactive with the reagent of Rydon but, for the preparation of the compounds of the invention, Ac and R are as given previously.

   For example, when

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 when a compound of formula IIA (Ac in formula II is that of the acid of formula A) is used as the starting compound, a new compound of formula IA is obtained. When this compound or the starting compound (formula IIA) is hydrogenated with a catalyst effective to saturate a clefinic double bond, a compound of the formula IB is obtained in the form of a mixture of the dis and trano epimers according to the * formulas :
 EMI4.1
 These epimers can be, if desired, separated by countercurrent ion diatribute or chromatography.



   When R3 in formulas B, IB and IIB is hydrogen, it can be replaced by suitable alkylation or a similar process. Advantageously, this replacement is carried out by reacting the compound according to formula B, IB or IIB, where R3 is hydrogen, with an oxo compote (an aldehyde or a cetane) and hydrogenating the product d. The resulting addition with a catalyst effective to cure a final olefin double bond. Platinum or palladium can be used as a catalyst. Suitable oxo compounds have the formula R4R5CO where R4R5C = is the same as R2 cited above.

   Examples of suitable oxo compounds are; formaldehyde, acetaldehyde, propionaldehyde, butyraldehyde, acetone, isobutylmethyl ketone, benzaldehyde, phenylacetaldehyde, hydrocinnamaldehyde, acetophenone, propiophenone,

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 butyrophenone, 3-methyl-4-phenyl-2-butanone, 2-methyl-5-phenyl-3-pentanone, 3-cyclopentanepropionaldehyde, cyclohexaneacetal-
 EMI5.1
 dehyde, cycloheptanecarboxalddhyde, 2,2-dimdthyicyâloprflpylactaldehyde, 2,2-imethylcyclopropyl methyl ketone, cyclopntyl methyl ketone, cyclobutyl methyl ketone, cyclobutanone, cyclohexanone, 4-methylcyclohexanone, etc.



   The starting compounds of formula II are prepared by acylating a compound of the formula:
 EMI5.2
      in which R 'is as given above, with an L-2-pyrrolidinecarboxylic acid substituted in position 4 of formula A or B.

   This acylation and similar acylations referred to herein can be achieved by methods already well known in the art for the acylation of amino sugars. The starting acid of formula A can be prepared by reacting a 4-oxo compound of formula
 EMI5.3
 
 EMI5.4
 wherein Z is a protective hydrocarbyloxyaarbonyl group which is separable by hydrogenolysis, trityl, namely triphenylmethyl, diphenyl (p-methoxyphenyl) methyl, bis- (p-methoxy-

  <Desc / Clms Page number 6>

 phenyl) phenylmethyl, benzyl or p-nitrobenzyl, with a reagent
 EMI6.1
 from Wittig for example an alkylidenetriphényiihosphorane see for example Wittig et al., Ber., 87, 1348 (1954) Tripett,

     Quarterly Reviewe, XVII, No 4, p. 406 (196317. Examples of hydrocarbyloxycarbonyl groups (Z) are tertiary butoxy carbonyl; benzyloxycarbonyl groups of formula 1
 EMI6.2
 in which X esthydrogen, nitro, methoxy, chlorine ou brama,
 EMI6.3
 for example carben, zaxyt p-nitrocarbobenzoxy, p-omo- and pc.ofsoarbabzay; and phenyloxyoarbonyl¯ groups of formula!
 EMI6.4
 where X1 is hydrogen, allyl, or alkyl of 4 ato-
 EMI6.5
 at most dp aarbone, such as phenyloxycarbonyl, p-tolyloxyarboy: e, p-6ylphenyloxycarbonyl and p-allylphenyloxycarbonyl. cte.



   In the implementation of this method, we add
 EMI6.6
 4-Oxo-L-1-pyrrolidinecarboxylic acid (formula C) to a freshly prepared Kairtrq reagent. The Wittig reagents used herein can be generally represented by the
 EMI6.7
 following formula! R1P (C6H5) 3 in which R1 is as given previously. These reagents
 EMI6.8
 this i.tzg are prepared by reacting an alkyl-, cycloalkyl- or aralkyltripënylphosphonima halide with a base,

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 such as sodium amide, or sodium or potassium hydride, or sodium or potassium metalate of dimethyl sulfoxide, etc.

   For example, the removal of hydrochloric acid from the alkyltriphenylphosphonium halide produces an alkylidenetriphenylphosphorane. The preparation of the phosphoranes is discussed in detail by Trippatt, Quart.



    Rev. XVII, No 4, p. 406 (196317. The reaction is generally carried out in an organic solvent, such as benzene, toluene, ether, dimethyl sulfoxide, tetrahydrofuran, etc., at a temperature between 10 ° C. and the reflux temperature of the mixture. reaction mixture.

   The product thus obtained, namely a 4-alkylidene-, 4-cycloalkylidene-, or 4-aralkylindene-1- protected-L-proline, which has the following formula
 EMI7.1
      is recovered from the reaction mixture in a routine manner generally by extraction from aqueous solutions of the reaction mixture. The crude product can be purified by usual means, for example recrystallization, chromatography or formation and formation. recrystallization of easily formed derivatives, such as the amino salts of the amino acid, for example the dicyclohexylamine salt, etc., and release of the amino acids from these compounds.

   By hydrogenation of an acid of formula B in the presence of a catalyst, for example platinum, which is effective in saturating a double bond, but which is ineffective in effecting hydrogenolysis, a compound of the following formula is obtained :

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 EMI8.1
 Platinum deposited on a support, for example carbon or an anion exchange resin, such as Dowex-1, which is a polystyrene trimethylbenzylammonium resin crosslinked in the hydroxide ring, is suitable. If desired, the starting compounds of formula II can be acylated with acids of formula C, D or E, to form compounds IIC, IID and IIE respectively.

   Compound IIC can then be converted to compound IID by treatment with a Wittig reagent, and compound IID can be hydrogenated to compound IIE by the methods given above. Hydrogenation of both D acid and IID acylate gives a mixture of cis and trans epimers which, if desired, can be separated by countercurrent distribution or chromatography. The starting acids of formula B, where R3 is hydrogen, are obtained when an acid of formula D or E is subjected to hydrogenolysis over a palladium catalyst, for example palladium on charcoal.



  Likewise, compounds of formula IID and IIE are converted to compounds of formula IIB, wherein R3 is hydrogen, by the same method. The starting acids of formula B, in which R3 is hydrogen, as well as the compounds of formula IIB, in which R3 is hydrogen, can be converted to the compounds of formulas B and IIB, respectively, wherein R3 is HR2 by the methods given above.



  The starting acids of formula A are obtained by treating an acid of formula D or formula E with acid

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 EMI9.1
 brom., hydric in acetic acid to separate the Z group, and then replacing the N-hydrogen with an HR2 group. the method given above. Compounds of formulas IID and IIE are converted to compounds of formula IIB by the same process.



   Some of the starting compounds of formula II are obtained by biosynthesis. Lincomycin, namely 6,8-dideoxy-
 EMI9.2
 Methyl 6- (trans-1-methyl-4-prppyl-L-2-pyrrolidinecarboxamido) -1-thioD-erythro-aD-galacto-octopyranoside, is obtained as a construct of an actinomycete producing lincomycin, according to US Pat. No. 3,086,912. It, the following structural formula:
 EMI9.3
 wherein R and R3 are methyl and R1H is propyl.

   La ', lincomycin B, namely 6,8-dideoxy-6- (trans-1-methyl-4-ethyl-
 EMI9.4
 Methyl L-2-pyrrolidinearboxamido) -1-thio-D-erythro-aD-galacto-oato-pyranoside (formula VI, in which P and R are methyl and -R1H is ethyl) is also a product of Preparation of the same lorr microorganism from a culture according to the method developed in US Pat. No. 3,086,912.

   Lincomy- (S-ethyl-S-demethyllincomycin),
 EMI9.5
 cine C namely 6,8-dideoxy-6- (trans-1-methyl-4-propyl-L-2-pyrrolidinecarboxamido) -l-thio-D-erythro-aD-galacto-octopyranoside ( formula VI where R is ethyl, -R1H is

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 propyl and R 3 is methyl) is obtained when the process according to U.S.A.

   n '3086912 is implemented in the presence of a
 EMI10.1
 addition of ethionine. lincoinycin D, namely 6, B-dideoxy-6- (trans-4-propyl-L-2-pyrrolid inecarboxamido) -1-thio-D-erythro-a-Dgalacto-oct., Methyl yranoside (formula VI in which R is methyl, -R1H is propyl and R3 is hydrogen) is obtained when the fermentation according to US Pat. No. 3086912 is carried out in the presence of an addition of a-MTL , namely 6-amino-
 EMI10.2
 Methyl 6,8-dieoxy-D-erychro-1-thio-a-D-gfllacto-octopyranoide, which is a compound obtained by the hydrazinolysis of lincomycin according to U.S. Patent 3,179,595.

   N-demethyllincomycin B, namely
 EMI10.3
 Methyl 6, 8-dideoxy-6- (trans-4-ethyl-L-2-pyrrolid inecarboxamido) -1-thio-D- erythro-aD-galacto-octopyranoside (formula Va / in which R is methyl, -R 1 H is ethyl and R3 is hydrogen) is also produced when α-MTL is added to the following fermentation: US Patent 3086912.

   Likewise, lincomycin K, namely 6,8-,
 EMI10.4
 dideoxy-6- (trans-4-propyl-L-2-pyrrolidinecarboxa! nido) -l'-thio-D-! Ethyl erythro-α-D-galacto-octopyranoside (Formula VI where R is ethyl, -R1H is propyl and R3 is hydrogen) is produced when the fermentation according to U.S. Patent 3,086,912 is
 EMI10.5
 carried out in the presence of an addition of ethyl α-BTL, namely α-amino-6,8-, d.cesoxy-I3-erythzoa ihio-D-galaata-oatopyranoside of ethyl, i.e. a compound obtained by the hydrazinolysis of lincomycin C. The S-
 EMI10.6
 , thy1-5,; - didëmdthyllxncomycin B, namely 6,8-dideoxy-6- (trans-; -éthy:

  .-. L-2-Fyrrol id inecarboxamido) -1-thio-T! -Erythro-a-D-galaato-octopyranoside (formula VI in which: R is ethyl,
 EMI10.7
 -R 1H is ethyl and R3 is hydrogen), is also obtained when α-ETL is added to the fermentation according to U.S. Patent No. 30S6912. The N-deamethyl products described <1-desaus, which are obtained * when adding α-MTL and Ó-ETL in the fermentation process according to U.S.A.

   3086912, are examples of starting compound IIB, in which R3 is hydrogen which, by

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 described
 EMI11.1
 process / previously, can have the X-hydrogens replaced when it is desired that R be equal to HR2, for example when producing P-3
 EMI11.2
 Desires 6,8-dideoxy-6- (trans-1-ethyl-4-propyl-L-2-pyrrplidin-oarboxamido) -1-thio-D-rhythmro-aD-galacto-datopyranosids fe methyl or 6, S-dideoxy-6- (trans-1-methyl-4-ethyl-L-2-pyrrolidine- carboxamido) -1-thio-D-erythro-GD-qalacto-ootopyranoside 'ethyl:

   or ethyl 6,8-dideoxy-6- (trans-1-thyl-4-ethyl-L-2-pyrrolidiriecarboxamidoj-1-thio-D-erythro-GD-galaato-octopyranoside or 6,8-dideoxy -6- (trans-1-ethyl-4-eflyl-L-2-pyrrolidinecar-) boxamide) -l-thio-D-èrythro-aD-galacto-octopyranoside, methyl,
Lincomycin or any of the starting compounds of formula II, which have the D-erythro configuration,
 EMI11.3
 can be converted to the L-threo configuration by oxidizing the 7-hydroxy group to a 7-oxo group and then reducing the latter to a 7-hydroxy group.

   A suitable process for this is illustrated by the following series of reactions:
 EMI11.4
 

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 EMI12.1
 For example, lincornycin, when treated with acetone in the presence of p-tolu.neau1fon1que acid, aat converted to 3,4-0-isopropylidene incomycin which, by oxidation
 EMI12.2
 with ahromic oxide, donna / 7 = dehydro3r4-p-iaopropylidenalincomycin (6,8-dideoxy-3,4-o-isopropylidene-6- (trana-1methyl-4-propyl-L-2-pyrrolidinecarboxamido) - methyl l-thio-D-glycérc> - * D-galaeto-octanopyranos-7-uloside (methyl), which on treatment with aodium borohydride is converted into 7-epilincomycin, 8-dideoxy-6- (trana-1 Methyl -methyl-4-propy, -hZ-pyrralidinacar-boxamido) -1-thio-L-threo-aD-galacto-octopyranoside.

   Any of the starting compounds of Formula II having a D-erythro configuration can be converted to the corresponding D-threo configuration by this method.



   Like lincomycins produced by biosynthesis,
 EMI12.3
 just as the amino-sugars derived therefrom are methyl or ethyl thioglucidons, it is sometimes desirable to convert them.
 EMI12.4
 shooting at higher or lower glvcosides. It is sometimes also desirable to convert any of the compounds of formulas I, II or V to higher or lower glycosides,
This can be done effectively by reacting the.

   compound to be converted with a mercaptan of the formula R6SH, wherein R6 is an alkyl group of 20 carbon atoms at the most, but an alkyl group other than R, for example compounds of formula I or II, upon reaction with a mercaptan of
 EMI12.5
 formula R6SH, produce thioacetals having formulated them
 EMI12.6
 

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 formulas in which X represents the hydroxy radical or a halogen and which, on treatment with an acid and / or by heating, are recycled to give a compound of the following formula '
 EMI13.1
 
The method can be applied directly to any of the starting * material * of formula II, i.e. IIA, IIB, IIC,

     IID and IIE. The resulting products can be subjected to hydrazinolyae to form compounds of the following formula:
 EMI13.2
 which can be N-acylated as described above, with acids of formula A, B, C, D and E, to give compounds according to formula XII, in which X represents the hydroxyl radical. The process can also be applied to the starting compounds of formula V. For example, α-MTL, by treatment with ethyl mercaptan, followed by recycling as previously described, is converted to α-ETL.



   Another method of preparing compounds of formula XI or formula XIII is to brominate the material.

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 starting and then reacting it with the mercaptan following the following succession of reactions:
 EMI14.1
 
The starting compound, XIV, is dissolved in water as a soluble salt, for example the hydrochloride, and bromine is added with cooling advantageously to a compiled temperature of between about -10 to 20 C.

   It suffices that the aqueous solution is cooled to about 0 ° C. and the bromine is added dropwise. The stoichiometric amount of bromine is 1 mole for each mole of the starting compound, although one can. use more or less. Advantageously, a slight excess, for example 5 to 20% excess, of the bromine is used.

   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 an acid, for example acid

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 hydrochloric acid or another strong non-oxidizing acid, such as p-tolueneulfonic acid, and sulfonic acid type anion exchange resins, mercaptan R6SH reacts with XV sugar to form thioglucoside XVI.

   At the same time, some of the diacetal of formula XI described previously after separation can be cyclized as described previously to form an additional amount of the desired thioglucoside XVI.



   The mechanism by which Rydon's reagent effects the substitution of the 7-hydroxy group by halogen is not fully understood. The mechanism, however, is believed to be such that a change in configuration results. Thus, a 7-hydroxy compound of the D-erythro configuration would give a 7-halo compound of the L-threo configuration.



   Rydon's reagents are formed by the addition of a halogen to triphenylphosphine or a triphenylphosphite or by the addition of an alkyl halide to triphenylphosphite, and can be represented by the formula
 EMI15.1
 

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   f'formules in which X is halogen, for example chlorine, bromine and. iodine (Rydon et al., J. Chem.Soc.



  2224 (1953) 1 ibidem, 2281 (1954), ibidem, 3043 (1956). The Rydon reagent can be formed in situ by adding a halogen or a methyl halide to a solution of triphenylphosphine or triphenylphosphite in an inert solvent, for example acetonitrile or dimethylformamide, or it can be isolated as a separate entity. In either case, the reaction with lincomycin or a related compound is carried out by contacting the Rydon reagent with this lincomycin or related compound, in an inert solvent, for example acetonitrile. or dimethylformamide, until the desired substitution of 7-hydroxy is obtained.

   The reaction proceeds at room temperature, although moderate heating can be carried out if delirious. Advantageously, the temperature is maintained between about 20 C and about
C. The product can be recovered from the reaction mixture by well known techniques, eg filtration, solvent extraction, etc. The reaction mixture is preferably treated with methanol to destroy any excess of Rydon reagent, then it is filtered to separate any solid, such as triphenylphosphine oxide, formed during the reaction. then it is processed to recover the product. Methanol can be added before or after filtration.

   The treated and filtered reaction mixture is preferably evaporated to dryness and purified by solvent extraction and / or chromatography.



   The compounds of formulas IA, IB, IIA, IIB and V exist in the protonated form or in the non-protonated form, depending on the pH of the environment. When the protonated form is considered, the compound is qualified as being a salt by addition of acid and, when the non-protonated form is involved, it is.

  <Desc / Clms Page number 17>

 qualified as the free base. The free bases can be converted to stable salts by adding acid, neutralizing the free base with the appropriate acid to below a pH of about 7.0, preferably up to about pH. from 2 to 6.

   Acids suitable for this purpose are: hydrochloric, sulfuric, phosphoric, thiocyanic, fluosilic, hexafluoroarsenic, hexafluorophosphoric, acetic, succinic, citric, lactic, maleic, fumaric, pamoic, cholic, palmitic, mucic, camphoric, glutaric , glycolic, phthalic, tartaric, lauric, stearic, salicylic,
 EMI17.1
 3-phenylsalicylic, 5-phényloalicylique, 3-methylglutaric, orthosulfobenzolque, cyclohexanesulfamic, cyclopentaneprop0- nique, 1,2-cyclohexanedicarboxylic, 4-cyclohexane, octadecenyl, octenyl succinic, methanesulfonic, benzenesulfonic, hqlianthique, Reinecke, dimethyldithiocarbamic, cyclohexylsulfamic, hexadécylsulfamique,

   octadecylsulfamic, sorbic, monochloroacetic, undecylenic, 4'-
 EMI17.2
 4-hydroxyazobenzene, octyldecyl, sulfuric, picric, benzoic, cinnamic and similar acids,
The acid addition salts can be used for the same purposes as the free base, or they can be used to improve the latter.

   By way of example, the free base can be converted to an insoluble salt, such as picrate, which can be subjected to purification processes, for example solvent extractions and washes, chromatography, liquid fractional extractions. liquid, and crystallization, and it can be used to regenerate the free base by treatment with an alkali or to achieve a
 EMI17.3
 different salt by metathene.

   The free base can also be converted to a water soluble salt, such as the hydrochloride or sulfate, and the dunsel aqueous solution can be extracted with various water-immiscible solvents, before regeneration of the free base form. by treatment of the acid solution

  <Desc / Clms Page number 18>

 thus extracted, or converted to another salt by metathesis. The free bases of formulas IA.IB, IIA, IIB and V can be used as buffers or as antacids. The compounds of formulas I, II and V react with isocyanates to form urethanes and can be used to modify polyurethane resins. Long chain compounds, that is, in which HR2 is an alkyl of 8 carbon atoms and more, have surfactant properties,

   and they can be used as wetting and emulsifying agents. The salt by addition of thiocyanic acid, when condensed with formaldehyde, forms resinous materials of value as pitting inhibitors according to US Patents 2425320 and '2606155. free bases are also good vehicles for! toxic acids, for example salts by adding acid! . fluosilicic acid are useful as anti-moth agents according to U.S. Patents 1915334 and 2075359, and addition salts of acids! hexafluoroarsenic and hexafluorophosphoric acid are useful as parasiticides according to U.S. Patents 3122536 and 3122552.



   Analogs close to 7-halo-7-deoxylincomycin, that is to say in which -R1H is cis or transalkyl of at most 8 carbon atoms, R3 is methyl or ethyl, R is a alkyl of up to 8 carbon atoms have antibacterial properties and some are comparable. or greater than lincomycin and can be used for any purpose other than lincomycin. Corresponding compounds where R3 is hydrogen have similar antibacterial properties, and in addition they have improved gram-free activity. .

   The other analogs and isomers have similar antibacterial properties, but to a lesser degree and can be used for the same purposes as lincomycin when larger amounts are not detrimental. The following examples illustrate the process and products of the present invention, but they do not constitute a limitation thereof.

   Parts and percentages are given by weight

  <Desc / Clms Page number 19>

 and the proportions of solvent are given by volume unless otherwise indicated. EXAMPLE 1 A, 7-bromo-7-deoxylincomycin and its hydrochloride
A solution of a Rydon reagent is prepared by stirring a dry solution of 52.6 gr (0.2 mol) of triphenylphosphine and 800 ml of acetonitrile at 30 under nitrogen, with the addition of 10 ml (0, 19 mole) of bromine over a period of 20 minutes. After stirring for a further 10 minutes, 8.2 g of lincomycin are added and the reaction is stirred at 30 for 18 hours. A white solid is then present. The reaction is filtered and the solid is discarded. Methanol (100 ml) is added to the filtrate and the solvents are then evaporated in vacuo.

   The viscous residue is dissolved in 100 ml of methanol, diluted with 1800 ml of water and extracted 6 times with 200 ml portions of ether. The ethereal extracts are discarded, the aqueous phase is made basic (pH 11) with aqueous KOH and then extracted 4 times with 200 ml portions of methylene chloride. The extracts are dried and evaporated, yielding 11 gr of a yellow solid which is chromatographed on a. kilo of silica gel using methanol-chloroform (1/9, v / v) as the solvent system. After a production head of 1200 ml, 22 fractions of 56 ml are collected. The last 6 fractions (fractions 17-22) are combined and evaporated to dryness, which gives 2.8 g of 7-bromo-7deoxylincomycin.

   This is converted to the hydrobromide by dissolving in water, adding HBr to pH 1, and filtering and lyophilizing the filtrate. The hydrobromide has an aD value = +114 (ci 0.9314, H2O) and the following analysis: Calculated for C18H34Br2N2O5S:
C: 39.28; H: 6.231 N: 5.091 S: 5.83t Br: 29.04 Found Ci 39.64, H: 6.191 N: 5.07; S: 6.04 Br: 28.59.

  <Desc / Clms Page number 20>

 



   Instead of bromine, other halogens can be substituted. Chlorine, for example, gives 7-chloro-7-deoxylincomycin which is identical to the product obtained by chlorinating lincomycin with thionyl chloride. Instead of triphenylphosphine, triphenylphosphite can be used. Further, in this case, a methyl halide can be used instead of a halogen. Instead of lincomycin, other lincomycins and their analogues can be used. Thus, when lincomycin C is substituted for lincomycin, 7-bromo-7-deoxylincomycin C.



  B. Preparation of lincomycin C.



   Lincomycin C is obtained by reacting lincomycin with ethanethiol (ethyl mercaptan) to form diethyl dithioacetal, followed by heating in the presence of p-toluenesulfonic acid or fusion. The following process is given by way of illustration.



  B1 6,8-dideoxy-6- (trans-1-methyl-4-propyl-L-2-pyrrolidinecar-boxamido) -D-erythro-D-galacto-aldehydo-octoea diethyl dithioacetal.
 EMI20.1
 



   150 cm3 of concentrated hydrochloric acid, and 50 cm3 of ethanol (previously cooled to 0), then 15.0 g of lincomycin hydrochloride, are placed in a bottle with a capacity of 1 liter, at 3 cola, After stirring magnetic at temperature

  <Desc / Clms Page number 21>

 At room temperature for 5 hours, the reaction mixture is diluted with an equal volume of ice-water and the solution is extracted.

   thoroughly with Skellysolve B (technical hexane), these extracts being rejected,
Most of the acid is neutralized by the addi-! Carefully add solid caustic potash (100 g), keeping the temperature of the reaction mixture well stirred, between 20 ° and 30 ° C., by cooling in acetone-dry ice. Solid potassium chloride is filtered off and the solid washed well with chloroform. An additional quantity of chloroform is added to the filtrate (about 150 cm 3) and the mixture, stirred magnetically, is adjusted to pH 10 by the addition of aqueous caustic soda (2N).

   The layer at is separated,. chloroform, the aqueous layer is extracted thoroughly with duthloroform, the combined extracts are washed twice with water and dried over anhydrous sodium sulfate. Separation of the solvent at 30 ° C. under vacuum gives a semi-solid residue which, when crystallized from acetone, gives 5.41 g of 6,8-dideoxy-6- (trans-1-methyl-4 -propyl-L-2-pyrrolidinecarboxamido) -D-erythro- D-galacto-aldehydo-octose diethyl dithioacetal in the form of colorless flattened needles, melting point 130-132 '.



  Concentration of the mother liquors gives an additional amount of material (1.50 g), with a melting point of 129-131 (total yield 6.91 g, 42.4%).



  Analysis: Calculated for C21H4206N2S2
C: 52.25; H: 8.77; N: 5.81: S: 13.29%
Found C: 52.38; H: 8.71; N: 5.93: S: 13.46% B2. Cyclization to lincomycin C.



   (a) Part of the diethyl dithioacetal from part B1 and part of p-toluenesulfonic acid monohydric are refluxed in 25 parts of acetonitrile until significant antibacterial activity is obtained. The reaction mixture is cooled and evaporated to dryness.

  <Desc / Clms Page number 22>

 and chromatographed on silica gel using a solvent mixture comprising ethyl acetate, acetone and water, in the ratio of 8/5/1. Fractiona 102 to 131 show antibacterial activity.

   Among these fractions, fractions 105 to 125 are combined, evaporated to dryness and crystallized from acetone acidified with hydrochloric acid, and recrystallized by dissolving in water and adding acetone to give crystals. hydrochloride. lincomycin C, melting point '149-153'.



   (b) The diethyl dithioacetal from part B1 is heated at 260 for about 3 minutes and the odor of ethyl mercaptan is noted. The product, on chromatography as in part B2 '(a), gives lincomycin C.



  C. Preparation of lincomycin C by fermentation
Incomycin C hydrochloride is prepared as follows
FERMENTATION
A lying soil culture of Streptomyces lincolnensis var. Lincolnensis, NRRL 2936 was used to inoculate a series of 500 ml Erlenmeyer flasks, each containing 100 ml of a seeding medium comprising the following ingredients:: Yeastolac (1) 10 gr Glucose monohydrate 10 gr NZ- amine B (11) 5 gr Tap water, quantity sufficient to make @ 1 liter (1) Yeastolac denotes a protein hydrolyzate of yeast cells (11) NZ-amine B denotes an enzymatic digestion casein from Sheffield.



   The pre-sterilization pH of the seed medium is 7.3. The seed material is grown for 2 days at 28 ° C on a rotary Gump knife operating at 250 rpm.

  <Desc / Clms Page number 23>

 a 5% inoculum of the seeding described above; (5 ml) is added to each of 30 500 ml Erlenmeyer flasks @ each containing 100 ml of the following fermentation medium:

   Glucose monohydrate 15 gr Starch 40 gr Melasaea Mêlasses 20 gr Wilson peptone liqueur n 159 (i) 10 gr Corn maceration liquor 20 gr Calcium carbonate 8 gr Bacon oil o, 5 ml City water, sufficient quantity to make . 1 liter (1) Wilson Peptone Liquor No. 159 is a preparation of enzymatically hydrolyzed proteins of animal origin
At the time of inoculation, Dl-ethionine is added to a final concentration of 2 mg / ml.



   The shaken flasks are collected after 4 days of fermentation at 28 ° C. on a rotary Gump shaker operating at 250 revolutions per minute. They titrate 200 mcg / ml on the test against S. lutea, which is described below. The total beer solids are about 20 mg / liter.



    PURIFICATION
The total beer (235 liters) from the DL-ethionine fermentation is filtered at harvest pH using a filter aid as needed. The mycelial cake is washed with water and this cake is then discarded. The filtered beer and the washing water (275 liters) are stirred for 45 minutes with 12.5 kg of activated charcoal :, and 2.5 kg of diatomaceous earth. The mixture is filtered and the filtrate is re- thrown. The charcoal cake is washed with 60 liters of water and the water is discarded. The cake is washed with 70 liters of 20% aqueous acetone and the resulting washing liquid is discarded. The cake

  <Desc / Clms Page number 24>

 is then eluted twice with 100 liter portions of 90% aqueous acetone.

   The eluates are combined (215 liters) and the solution is concentrated (18 liters). This concentrate is adjusted to pH 10.0 with a 50% aqueous solution of caustic soda, and is extracted 3 times with 20 liter portions of methylene chloride. The methylene chloride extracts are combined (60 liters) and then concentrated to give an oily repair (7.14 gr) containing lincomycin and lincomycin C in equal amounts and both form the free base. . This preparation is then dissolved in 200 ml of methylene chloride.

   The solution is clarified by filtration and then concentration to dryness in vacuo. The residue is dissolved in 100 ml of 1N methanolic hydrochloric acid. The methanolic solution is then mixed with 3.2 liters of ether with stirring. The resulting precipitated colorless crude lincomycin hydrochloride and lincomycin C hydrochloride are isolated by filtration and dried .. the yield is 7.14 gr grading 940 mcg / mg against Sarcina lutea (The test against Sarcina lutea -vis of Sarcina lutea is carried out on agar-agar buffered at pH 6-8 with phosphate buffer of pH 7.0 [0.1M].

   A unit volume [0.08 ml] of a solution containing the material to be titrated is placed on a 12.7 mm test disc which is then placed on an agar plate seeded with the micro- agar. test organism). Thin layer chromatography shows the presence of both lincomycin hydrochloride and liricomycin C hydrochloride in approximately equal amounts.



   The crude lincomycin C hydrochloride (7.0 g) is dissolved in 20 ml of water and 20 ml of butanol, the pH is adjusted to 4.2 with 1N HCl, and the solution is distributed in a measuring apparatus. distribution or distrubtion against the current for 1000 transfers. Analysis by thin layer chromatography

  <Desc / Clms Page number 25>

 shows that the fractions in tubes 135 to 190 contain lincomycin C. These fractions * are combined and the solution is concentrated and dried thoroughly to give 2.44 g of lincomycin C hydrochloride assaying 1400 mcg / mg against -vis of Sar- cina lutea. 500 mg of this preparation are dissolved in 2 ml of water, 1 ml of methanol and 100 ml of acetone. The solution is clarified by filtration.

   The filtrate is mixed with ether until crystals appear. The rapos mixture is left at room temperature for 1 hour. Crystalline lincomycin C hydrochloride (cubes) is separated from the supernatant solution by decantation. These crystals are recrystallized from 1 ml of water, 1 ml of methanol, 80 ml of acetone and 20 ml of ether? 7 the yield is 250 mg crystalline lincomycin C hydrochloride (cubes). The supernatant deterioration (obtained as described above) is left to stand at 5 ° C. for 4 hours.



   .



  The crystalline incoinycin C hydrochloride (needles), which precipitates, is filtered and dried; the production is 150 mgr of crystalline lincomycin C hydrochloride (needles), with a melting point of 151-157 C.



  D, Alternative method of preparing lincomycin C
Lincomycin hydrochloride (8.85 g - 0.02 mole) is dissolved in 20 ml of water, cooled to 0 and stirred, while bromine (3.52 g, 0.022 mole) is added. drip over a period of one minute. Ethanethiol (25 mL) is added and the mixture is stirred at 25 for 2 hours. The clear, colorless 2-phase system (ethanethiol is relatively insoluble in water) is cooled in. an ice bath and bubbled hydrochloric gas through it for about 5 minutes.



  The lower aqueous phase turns red. The reaction mixture is then extracted 3 times with 100 ml portions of Skelly-solve B and an aqueous solution of caustic soda is added to bring the aqueous phase to pH 11. The basic phase is extracted

  <Desc / Clms Page number 26>

 suitably with chloroform. The chloroform extracts are washed with saturated sodium chloride solution, dried and evaporated in vacuo to give 6.2 g of a white solid. 4.8 g of this solid are chromatographed on 800 g of a gel. silica by the process of Example 2, using methanol-chloroform (1/7) as the solvent system. After 800 ml of production head, 80 fractions of 25 ml each are collected.

   Fractions 40-58 are combined and evaporated to dryness and the residual solid is recrystallized from acetone to give 0.5 g of a material identical to the diethyl dithioketal of part B1. Fractions 65-75 are combined, evaporated to dryness and dissolved in a mixture of 5 ml of methanol
 EMI26.1
 and 400 ml of d.L-ethyl ether. Hydrogen chloride gas is added and the white solid which precipitates is collected. By recrystallization from aqueous acetone, 0.5 g of lincomycin C hydrochloride is obtained, which is identical to that of part C.
 EMI26.2
 



  E. Other 7-halo-6,7,8-trideoxy-6- (trans-1-methyl-4-propyl-L-2pyrrolidinecarboxamido) -1 -thia-aD-galaeto-octopyranosidee of alkyl ¯ ¯, ¯, ¯ By substituting the etianethiol of part B1 and part D of this example with other alkyl mercaptans, for example propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexa-
 EMI26.3
 deoyl, heptadecyl, octadecyl, nonadecyl, and eicosyl mercaptans and their forms inomerular by cycloalkyl mercaptans, for example cyclopropyl, cyclobutyl, cyclopentyl, cyclo'hexyl, cycloheptyl, cyclooctyl, 2-methylcyclopentyl, 2-methylcyclothyl, 2,3-dimëthyl methylcyclobutyl and 3-cyclopentylpropyl mercaptanst or by aralkyl.mercaptans, for example! benzyls,

   phenethylt 3-phenylpropyl and a-l-phthylmethyl mercaptans, one obtains the alkyl cycloalkyl and aralkyl, 8-dideoxy-6- (trane-1-methyl-4-pro

  <Desc / Clms Page number 27>

 
 EMI27.1
 pyl-L-2-pyrrolidineaarboxamido) -1-thio-D-erythro-a-D-galacto- .. octopyranosides corresponding which. particularly by the process of part A, are converted into the alkyl, cycloalkyl
 EMI27.2
 and aralkyl 7-balo-6,7 # 8-tridenoxy-6- (trans-1-methyl-4-prajpyl-L-2-pyrrolidinecarboraraido) -1-thio-α-D-galacto-octanopyxsoaides corresponding.

   The compounds thus obtained (both the compound '6,8-dideoxy and the compound 7-halo-6,7,8-trideoxy), in the
 EMI27.3
 which akyle is propyl, butyl, pentyl, and hexyl (obtained respectively when propyl, butyl, pentyl and hexyl mer-captans are used, are special antibacterial agents).
 EMI27.4
 which are effective, which have the same spectrum as * lincomy4ixe 'and' '"equal or greater activity.



   Further, when lincomycin is substituted with 6,8-dideoxy-6- (trans-1-methyl- or 1-ethyl-4-butyl-L-2-pyrrolidinecarboxamido) -1-thio-D-erythro-aD methyl-galacto-octopyranoside, the compound 7-bromo-7-deoxy corres-
 EMI27.5
 laying, namely 7-bromo-6,7,8i-trideoxy-6- (trans-1-methyl and 1-ethyl-4-butyl-L-2-pyrolidinecarboxamido) -l-thio-ct-D-galac - Methyl to-octopyranosides, having the same antibacterial spectrum as lincomycin and equal or greater activity.



   When we substitute the cis epimers, we get them!
 EMI27.6
 ? -bromo-6,?, 8-tridéscxy-6- (Cisyl-mdthyl and 1-ethyl-4-butyl-L-; 2-pyrrolidinecarboxamido) -1-thio-aD-galacto-octopyranosidea of methyl having the same spectrum antibacterial.
 EMI27.7
 The cie and trans epimers, used as starting materials in the previous examples, were prepared as follows.
 EMI27.8
 



  4-butylidne -, - carbobenxoxy-L-proline and its cyclohexylamine salt, ¯¯¯¯, ¯.



   Sodium hydride (19 gr) in the form of a 53% suspension in mineral oil is heated with 350 ml of dimethyl sulfoxide at a temperature of 70-75 C, until

  <Desc / Clms Page number 28>

 that the reaction is complete (about 30 minutes). After cooling to 32 ° C, 16.2 g of butyltripenuylphosphonium bromide is added, and the resulting reaction mixture is stirred for 1 hour to ensure complete reaction. A solution of 26 g of 4-keto-1-carbobenzoxy-L-proline in 100 ml of dimethyl sulfoxide is added and the resulting reaction mixture is heated at 70 C for 3 hours.

   The reaction mixture is cooled to 25 ° C and 1 liter of a 2.5% aqueous solution of potassium bicarbonate is added. This mixture is washed twice with 700 ml portions of ether and the ether is discarded after exhaustion with 150 ml of a 2.5% aqueous solution of sodium bicarbonate. The bicarbonate solutions are combined and acidified with 4N hydrochloric acid. The acidified mixture is extracted with 4 portions of 500 ml of ether. The combined ethereal extracts are washed successively with 250 ml of water, 3 portions of 250 ml of saturated aqueous sodium bisulfite and 250 ml of water, then dried over anhydrous sodium sulfate. Evaporation of the solvent in vacuo gives 24 g of an oily residue which is 4-butylidene-1-car-bobenzoxy-L-proline.



   This residue is dissolved in 31 ml of acetonitrile and treated with 18 ml of dicyclohexylamine, then refrigerated. The crystals are collected, washed with acetonitrile and dried in vacuo to give 21 g (46.8%) of the crystalline dicyclohexylamine salt, melting at 136-140 ° C. After two recrystallizations from water. acetonitrile, an analytical sample is obtained which melts at 142-144 C and has a rotation of [Ó] D -4 (c = 0.99, CHCl30.



  Analysis: Calculated for C29H44N2O4
Ci 71.861H: 9.15; N: 5.78
Found here 71.691H: 9.30; Ni 5.74
10 gr of the dicyclohexylamine salt of 4-butylidè

  <Desc / Clms Page number 29>

 ne-1-carbobenzoyx-L-proline are shaken with ether and 5% excess of aqueous caustic potash until no solid remains. The layers are separated and each is washed back. The aqueous alkaline layer is combined with the backwash from the ethereal layer and acidified with 4N hydrochloric acid.

   The mixture is repeatedly extracted with ether and the combined ethereal extracts, dried over sodium sulfate and evaporated in vacuo to give 6.3 gr (93%) of 4-butylidene-carbobenzoxy-L-proline. in the form of an oil.



   G. 4-butyl-1-carbobenzoxy-L-proline
The oil from part F is hydrogenated in 200 ml of methanol over 2.1 g of a 10% platinum catalyst over Dowex-1, under a pressure of 40 pounds of hydrogen. The catalyst is separated by filtration and the filtrate is evaporated to give 6.3 g of 4-butyl-1-carbobenzoxy-L-proline in the form of an oil. The product contains about 2 parts of cis-4-butyl-1-carbobenzoxy-L-proline for each part of trans-4-butyl-1-carbobenzoyx-L-proline.



   If desired, the hydrogenation of the 4-ylidene group can be postponed to any subsequent phase, even the final phase, in the process.



   By substituting butyltriphenylphosphonium bromide of part E by other substituted triphenylphosphonium bromides, in which the 4-substituent is methyl, ethyl, propyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, heptadecyl, octadecyl, nonadecyl, and eicosyl and their isomer forms; cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, .2-cyclopropylethyl and 3-cyclopentylpopyl;

   benzyl, phenethyl, 3-phenylpropyl and a-naphthylmethyl,

  <Desc / Clms Page number 30>

 
 EMI30.1
 the 4-alkylidene 4 r ,, talky.idene-, and 4-aralkylidene-1-carbobenzoxr-L-pro, + ei. corresponding. The 4-alkyl-, 4cycloalkyl-, and 4-aralkyi-1 are obtained. corresponding abenzoxy-1, -prolin.

   For example, 10r8qu51f.butyltriphenylpholphonium iromide is substituted by ethyl-, propyl-, isobutyl-, pentyl-, and hexyltrfinf1phosphonium bromides, 4-ethylidera-1-carbobenzoxyyoline, 4-propylidene-1- is obtained. carbobenzoxy-L-proline, 4-isRiYlidànè-l-carbobenZOXY-L-prolin., 4-pentylidene-1-carbobenzgI-proline, and 4-hexylidene-1carbobenzoxy-L-proline and cis and trans 4-ethyl -l-carbobenzoxy-h-prolin3s, 4-propYl-li! {obenzoxY-L-proline, 4-isobutyl-1-carbobenzoxY-L-prolinted + a 4-pentyl-1-carbobenzoxy-Lproline, and 4 -hexyl-1-carbobenzoxy-L-proline.



   H. Methyl 6-amino-6,8-dideoxy-1-thio-D-erthro-Ó-D-galacto-octopyranoside (Ó-MTL).



   A solution of 40 g of the free base of lincomhydrate cine (U.S. Patent No. 3,086,912) in 20 ml / hydrazine (98-100%) is refluxed for 21 hours! the excess hydrazine hydrate is then removed in vacuo under nitrogen, the temperature of the steam bath, leaving a residue. The residue, which a pitiful mass of crystals, is cooled, acetonitrile is added and the mixture is stirred until the crystals are suspended. The crystals are collected on a filter, washed with acetonitrile and with ether.

   The yield of white crystalline α-MTL free base after vacuum drying at room temperature is 21 g (84%). Recrystallization is carried out by dissolving the free base of α-MTL in hot dimethylformamide and adding an equal volume of ethylene glycol dimethyl ether.



   Methyl 6-amino-6,8-dideoxy-1-thio-D-erythro-aD-galacto-octopyranoside free base has a melting point of 225-228 C, an optical rotation of [Ó] 25 = +276

  <Desc / Clms Page number 31>

 
 EMI31.1
 (c - 0.768, water) and a pKa 'value of 7.45.
 EMI31.2
 Analysis 1 Calculated for C 9 H 19 NO 5 S
 EMI31.3
 
 EMI31.4
 CI 42, 7 t.

   F3: 7.561 Nt 5.531 Si 12.66, Found Ci 42.6; to 7 <49r Nt 5, 75 Si 12, 38 By substituting lincomycin by others 6 8diddsoxy-6- (trans-1-methyl-4-propyl-L-2-gyrrolidinecarbaxamido) - 1-thio-D-drythro-aD -alkyl galacto-octopyranosides, cyclo-
 EMI31.5
 alkyl or aralkyl, in which the akyl is ethyl, propyl,
 EMI31.6
 butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, lundéey3e dodecyl, trideayl, tetradecylei pentadecyl, hexadecyl,
 EMI31.7
 
 EMI31.8
 hoptaddeyle, octad6ayl., nonadéayle, and eic08yle and their forms i8o'r.l, cycloalkyl is:

   t cyolopropyl, cyclobutyl, cyclopentyl, cyclohoxyl, cycloheptyl, cyclooctyl, 2-methylayalopentyl, 2,3-dimethyicyciobutyl, 2-mithyloyolobutyl, and 3cyclopentylpropylel and aralkyl is 1 benzyl, phenethyl, 3-
 EMI31.9
 phenylpropyl, and a-naphthylmethyl, we obtain the alkyl, ring- '
 EMI31.10
 corresponding alkyl and aralkyl 6-amino-6,8-diddaoxy-1-thia-D-erythro-a-Dgalacto-octopyranoside.

   For example, substituting lincomycin with ethyb, propyl-, butyl-, pentyl-, and hexyl 6.8-dideoxy-6-trans-1-metriyl-4-propyl-L-2-pyrrolid.necarboxamido) -1-thio -D-erythro-aD-galaato-octopyranosidea, we obtain 6-amino-6,8-dideoxy-1-thio-D-erythro-aD-galacto-octopyranoside of ethyl, 6-amino-6,8- dideoxy.l-thio-D-ery-
 EMI31.11
 propyl thro-a-D-galacto-octopyranoside, 6-amino-6,8-
 EMI31.12
 butyl dideoxy-1-thio-D-erythro-dD-galacto-octopyraposide, butyl 6-amino-6,8-dideoxy-1-thio-D-erythro-aD-galacto-octopyranoside and pentyl 6, amino- 6,8-dideoxy-1-thio-D-erythro-aD-
 EMI31.13
 hexyl galacto-octopyranoside.
 EMI31.14
 



  If desired, the method of Part A can be applied to the compounds of Part H using the hydrochloride or other salt of a fore acid, and the resulting 7-halo compounds can be treated by the following steps of this

  <Desc / Clms Page number 32>

 example to arrive at the final products of the same example.
 EMI32.1
 .

   B, 8iideox6 free base (.waa, rbobenocy-4 bul-h - 2-isYrrl id i nensoxanait o) -.- th io: D-é-. thro-Q-D-1.9to-9gtOPVTno.ide from m6tbvl "'. @
 EMI32.2
 
 EMI32.3
 3.46 ml of triethylamine are added to a solution of 6.3 g of 4-butyl-1-oarbopenzoxyL-proline (the oil from the Cri part in 175 ml of distilled acetonitri18, cooled to 010. then 3.34 ml of inobutyl chloroformate, The mixture is stirred at 0 ° C (t 3.) for 5 minutes, A solution of 6.2 g of the free base of a-MTL from part C in 85 ml of water is added and the reaction mixture is stirred at 0 C for 1/2 hour and at 25 C for 1 hour.

   The reaction product is then filtered and dried, yielding 4.57 gr (37.7%) of the
 EMI32.4
 6,8-Diddaoxy-6- (1-axxbobenaoxy-4-butyl-L-2-pyrroli-. dinecarboxarnido) -1-thio-D-erythro-α-D9alaoto-oct pyr & n08ide free base. The mother liquor is concentrated under vacuum and we recover
 EMI32.5
 4.25 additional gr of product (35.2%). Recristallization from llicetonitrile produces crystals of the free base of 6,8-deoxy-6- (1-carbobenzoxy-4-butyl-j-2-pyrrolidinecarboxamido) -1-thio-D-erythro-aD-galauto-octopyranosià methyl, melting at 194-196DC. A second recrystallization from acetonitrile gives the analytical sample, with a melting point of
 EMI32.6
 195.5-200 C, having a value of 7.D - + lll * (0 * O, 981 Mef? Fi.

  <Desc / Clms Page number 33>

 
 EMI33.1
 



  Analyae ' <Calculated for C26N40N20eS Ci 57.751 Es 7.46; Nt 5 # 13Y Si 5.93 Found Ct 57, SS, Es 7 # 161 Nt 5.501 Os 6.07 1 iY, 6,8-denoxy-6- (4-butyl-L-2-t hydrochloride). .



  PYr'rsZidinarboxxvmido) = 1-thi.an-erythroc-Da3.aato- eto methyl anoside '
 EMI33.2
 
 EMI33.3
 A solution of 7.8 gr of the free base of 6.8-
 EMI33.4
 dëao-- 1-ctrbobanzoxy-4-butyl-L-2-Pyrzolidinecarboxamido) - 1-thio-D-erythro-gD-galacto-oatopyranoside from part 1 in 200 ml of methanol is shaken on 2 gr of 'a palladium catalyst at 1056 on' carbon pressure us
 EMI33.5
 of 40 pounds of hydrogen for 17 hours. The catalyst is filtered off and the solution is concentrated in vacuo. The residue is dissolved in a mixture of 20 ml of acetone and 20 ml of water, then it is acidified with 6N hydrochloric acid.

   Dilution with 4 volumes of acetone precipitates the hydrochloride.
 EMI33.6
 Methyl 6,8-deoxy-6- (4-butyl-L-2-pyrrolidinecarboxanido) -1thia-D-erythra-α-D-galxcto-octapyranaeide drate, which is filtered off and dried. The crystals, dried at 550 C under vacuum, weigh 4,? gr and melt at 188-194 * C. The analytical sample obtained by acetone reaction melts at 197-199 OC and gives a ccc value of + iso. (water, c- t3.89.



  Analysis t Calculated for C 3 N O S, HCI 18 calCU16 for e Î034 "2068. Iiel Ci 48.801 Hi 7.961 Ni 6.321 St 7.24
 EMI33.7
 Found (corrected for 5.54% water)
 EMI33.8
 Cs 48.581 and 8.191 Ne 6.04; Si 7.36

  <Desc / Clms Page number 34>

 
 EMI34.1
 This material has 8% of the antibaotder activity of .3.aaomc3, ne vit-a-'vi * of 8, lutea.



  Zn substituting o-MTL by Other alkyl or cyclo-alkyl or aralkyl 6-a.no-6, 8-d.eoxy-l-thio-D-rybhro-a D-gal, aato-orta, topyranoeidas, wherein the alkyl is ethyl-propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, noadeyl, and eicosyl deciles in their 4 isomeric forms = cycloalkyl is oyolopropyl, ayeloutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyalooatyl, 2-methyir cyclopentyl, 2,3-dimethylcyclobutyl, 2-mithylcyclobutyl and 3-cyclopentylpropyl, and aralkyl is:

   ben2yle. phenethyl, 3-phenylpropyl, and 0-naphthylmdthyle, one obtains the alkyl, cycloalkyl, and aralkyl 6,8-diddsoxy-6- (1-carbcxbenzaxy-4-butyl-L- 2-PYrro1 id inecarboxanido? -l-thio- D-erythro-Gs-D-galacto-oatopyra
 EMI34.2
 
 EMI34.3
 acids and alkyl, cycloalkyl, and aralkyl 6,8-dideoxy-6- (4-butyi-L-2-PYrrolidinecarboxazai.do-1-thio-erythro-a-D-galacto-
 EMI34.4
 corresponding octopyranosides.

   For example, substituting the
 EMI34.5
 a-MTL by ethyl, propyl, butyl, pentyl, and hexyl 6-amino-fi; dideaoxy-1-thio-D-erythro-a-D-galacto-octopyranoaidea, it is obtained. 6,8..didenoxy-6- (ethyl 1-carbobanzoxy-4-butyl-L-2-pyrrolidintaaxboxamiso71-thie-D-erythra-o.D-galaoto-otopyranoide;

   6,8-didéaraxy-6: -caxbabarzcocy-4-buty.Lpyrrolidinesarbrrxa mido) -1-thio-D-ery% hro-aD-qalacto-octopyranoide propyl, 6,8 dideQxy-6- (1-aarbainxvy -4-Jantyâ.-L- pyrroiidinecarboxazai do) -1-thio-D-erythro-aD-qalac% o-octopyranoide butyl, 6, 6-dideaoxy-6- (1, -carbcberzexy-4-buty.- L- = pyrroXidinacrbararni do) -1-thio-D-erythro-aD-galacto-octpyranooid pentyl, 6, 8-dideaxy-6- (i-carbobnzmcy-r-buty .-- L2-pyx ralidinecarboxami do) - Hyl 1-thio-D-erythro-aD-galacto-octopyranooid, 6,8d ideaoxy-6- (4-butyl-L-2-pyrrolidmecarboxamido) -1-thio-D-erythrOa-D-galacto- ethyl octopyranoside, 6rdidstnty.ô 4-butyi-

  <Desc / Clms Page number 35>

 
 EMI35.1
 L-2-pyrrolidineaArboxamido) -1-thio-D-erythro-aD-qalacto-octopyranoside propyl, 6, 8-didiraxy-6 (4-butyl-L-2-pyrro, idlnor¯ aarbaxamido) -1-thio -D-erythro-al- ra,

  butyl acto-aatapyxanoside, pentyl G, 8-dideaoxy-â-t4-butyl-L-2-pyrxoliQinecarboxamido) -.-, thio-D-'rythro-aD-galaato-ootopyran08ide and 6,8d ideraxy-6 (4..butyl-L3-pyxrol id ineaarboxamido 'wl-th3, oD-erythroc.



  Hexyl D-galacto-octopyranoside.



  These alkyl, cycloalkyl and aralkyl L-thiolicolaminides obtained by hydrazinolysis of the corresponding 8-alkyli 6-cycloalkyl and 8-aralkyl S-demethyllincomycins' which, in turn, are prepared by substituting the ethanethiol of Preparation B-1 with
 EMI35.2
 other alkyl mercaptans, for example propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octodeoyl, nonadecyl and eicosyl mercaptans and their formsea isomers, by cycloalkyl mercaptans, for example cyclopropyl, iyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 2-methylcyclopentyl, 2,3-dimethylcyclobutyl,

   2-methylcyclobutyl and 3-cyclopentylpropyl mercaptans; or by aralkyl mercaptans, for example benzyl, phenethyl, 3-phenylpropyl and 1-naphylmethyl mer-captans.



   By substituting 4-butyl-1-carbobenzoxy-L-proline
 EMI35.3
 by other 4-alkyl-1-carbobanzoxy-L - prolines in which the 4-alkyl is: ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undéayl, dodecyl, tridecyl, tetradecy.
 EMI35.4
 le, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, et eicosyl and their isomeric forms; by 4-cycloalkyl-1le
 EMI35.5
 1 carbobenzoxy-L-prolines, in juelles / 4-cycloalkyle is cyclopropyl, cyclobutyl, cycloyl, oyolohexyl, cycloheptyl, cyclooctyle, 2-méthylcyaloptylo, 2,3-diméthylyclobutyle ,.



  2-methyloyolobutyl, and 3-cyclopertylprapyr ,, t by 4-aralkyl-

  <Desc / Clms Page number 36>

 
 EMI36.1
 1-oarbobenaoxy-L-prolinoa in which the 4-aralkylo is benzyl, phenethyl, 3-phenylpropyl and α-naphthylm4thyle <we skiatient alkyl, cycloalkyl and aralkyl 6,8-diddsoxy-6- (1-aarbobenzoxy-4-alkyl, 4-cycloalkyl 'and 4-aralkyl-L-2-pyrro11d1neoarbo- xamido) -1-thio-D- erythro-aD-qalaato-octopyranoaidea oorroapondants and alkyl, cycloalkyl and ralkyl- (4-alkyl, 4-cycloalkyl and 4-aralkyl-L-2-pyrrolid inecarboxamido) -1-thio-D-erythro-A-Dgalacto -octopyranooides corresponding, For example, by substituting 4-butyl-1-carbobeaoxy-L-proline by 4-methyl-, 4-ethyl-,
 EMI36.2
 4-propyl-, 4-pentyl-, and 4-haxyl-1-carbobenzoxy-L-prolines, we obtain methyl. ethyl.

   propyl, butyl, pentyl, and hexyl 6,8-
 EMI36.3
 dideoxy-6- (1-carhobenaoxy-4-methyl-L-2-pyrrolidinecarboxamido) - 3, -thio-D-erythro-aD-galacto-octapyranoaides F methyl, ethyl, propyl, butyl, pentyl, and hexyl 6,8 -c9ideoxy-6- (1-carbobenzoxy- 4-ethyl-L-2-pµrrolidinecarboxamido) -1-thio-D-erythro-AD-galaato-

  <Desc / Clms Page number 37>

 
 EMI37.1
 octopyranosides7 methyl, ethyl, propyl, butyl, pentyl and hexyl 6,8-dideoxy-6- (1-carbobenzoxy-4-propyl-L-2-pyrroliàinecarboxamido) -1-thio-D-erythro-aD-galacto-octopyranosidesi 'lfls methyl, ethyl, propyl, butyl, pentyl, and hexyl 6, 8-d. deoxy (1-
 EMI37.2
 carbobenzoxy-4-pentyl-L-2-pyrrolidinecarboxamido) -1-thio-Déry-
 EMI37.3
 thro-a-D-galacto-octopyranoides = methyl, ethyl, .propyl,
 EMI37.4
 
 EMI37.5
 butyl, pentyl and hexyl 6,

  8-dideoxy-6- (1-carbobenzoxy-4-he (yl-L-
 EMI37.6
 
 EMI37.7
 2-pyrrolidineaarboxamido) -1-thio-D-erythro-α-D-galacto-octÇpyranosides; methyl, ethyl propyl, butyl, pentyl and hexl 6,8didsoxy-6- (4-methyl-L-2-pyrrolidinecarboxamido) -1thio-D-ery-
 EMI37.8
 thxo-aD-galacto-octopyranosidesf methyl ,, ethyl, propyl, butyl] pentyl, and hexyl 6,8-dideoxy-6 - (- ethyl-L-2-pyrrolidinecarbaxa- 'midoi-1-thio-D-rhythmro- aD-galaato-oatogyranosides the meflyl,
 EMI37.9
 1
 EMI37.10
 ethyl, propyl, butyl, pentyl and hexyl 6,8-dideoxy-6- (4-propyl-L- 1 2-pyrrolidinecarboxamido) -1-thio-D-erythro-a-D-galacto-octopyra-
 EMI37.11
 
 EMI37.12
 noaidea;

   methyl, ethyl, propyl, butyl, pentyl, and hexyl 6, a-
 EMI37.13
 dideoxy-6- (4-pentyl-L-2-pyrrolidinecarboxamido) -1-thio-D-erythro-
 EMI37.14
 a-D-galacto-octopyranosides and methyl, ethyl, propyl, butyl,
 EMI37.15
 pentyl, and hexyl 6,8-dideoxy-6- (4-hexyl-L-2-pyrrolidinacarboxamido) -1-thio-D-eryfihro-a-p-galaoto-octopyranosides.



  If desired, the 1-carbobenzoxy compounds prepared according to part 1 can be halogenated by the process of
 EMI37.16
 part A and the resulting 7-halo compound can be treated by the following steps of the example to separate the 1-carbobenzoxy group and to substitute the proline nitrogen to give the end products of the example.
 EMI37.17
 



  Xl. 6,8-deoxy-6- (1-metlljl-4-butyl-L-2-pyrrolidinecarboxamido) -1-thio-D-eryNwo-a-D-galacto-octopy-
 EMI37.18
 methyl ranoside
 EMI37.19
 

  <Desc / Clms Page number 38>

 a solution of 2.0 g of 6,8-deoxy- hydrochloride
 EMI38.1
 Methyl 6- (4-butyl-L-2-pyrrolidinecarboxamido) -1-thio-D-erythro-aD-galacto-octopyranoside from the part and 2.0 ml of 37% formalin in 150ml of methanol is shaken on 500 mg of 10% palladium on charcoal under 40 pounds of hydrogen pressure for 3.5 hours. Separation of the catalyst by filtration and the solvent by distillation under
 EMI38.2
 vacuum gives 6,8-deoxy-6- (1-methyl-4-butyl-L- hydrochloride).



  . Partially crystalline methyl 2-pyrrolidinecarboxamido) -1-thio-D-erythro-aD-galacto-ootopyra- noside which by thin layer chromatography on gel, silica using a mixture of ethyl acetate, d acetone and water (8/4/1) for elution and a solution of KMnO4 for detection, turns out to consist mainly of two materials, namely the epimes-
 EMI38.3
 ras ci and trana of methyl 6 # 8-denoxy-6- (1-mithyl-4- 'butyl-L¯2-pYrroli8inecarboxaraida9-1-thia-ri-drythro-an-galacto-octopyranoside hydrochloride in a ratio of 'about 3 to 2.



     K2. Separation of cis and trans forms by chromatography
6,8-Deoxy-6- (1-methyl-4-butyl '' hydrochloride
 EMI38.4
 Methyl L-2-pyrrolidinecarboxamida-1-thia-T3-erythra-aD-galacto-octopyranoside from part K1 is dissolved in a mixture of methanol and methylene chloride (1/1) and added 1.5 ml of triethylamine. To this solution, 7 g of a silica gel and the solvent are added and evaporated in vacuo, which tires out the antibiotic deposited ', on the silica gel, which is separated at the top of a chromatographic column of 200 g of silica gel, packed with a solvent mixture consisting of ethyl acetate, acetone and water in a ratio of 8/4/1.

   The column is developed by elution with the same solvent and 20 ml portions are collected. Thin layer chromatography of each fraction shows that fractions 31 to 38 (310 mg) consist of the essentially pure trans epimer

  <Desc / Clms Page number 39>

 and that fractions 49 to 74 (32 mg) consist of the essentially pure cis epimer. Fractions 39 to 48 consisted of a mixture of epimers which could then be separated by repeated chromatography. Each epimer is dissolved in a few drops of dilute hydrochloric acid and the hydrochloride is precipitated by addition of acetone.

   In this way, we
 EMI39.1
 obtains 50 mg of 6,8-deoxl! -6- (trans-i-methyl ,, 4-butyl-L-2-pyrrolidinecarboxamido) -1-thio-D-erythro-tc-D-galacto-octopyranoside hydrochloride from methyl, melting point 135-1370, and about 150 mg of 6,8-deoxy-6-icis-1-methyl- ¯ 4-butyl-L-2-pyrrolidinecarboxamido) -1-thio hydrochloride Methyl-D-erythro-aD-galacto-octopyranoside, softening at 105 C with further melting at 175-185 C.



   The trans epimer recrystallized from the same solvent melts at '139-141 C and presents the following analysis:
 EMI39.2
 Analyae 1 Calculated for C 19 H 36 N 2 0 6 S.HC1 Ci 49.93y'Hs 8.61 Nt 6.131 Se 7.02 Found e (corrected for 4.07% H2n) Ce 48, Slr Hs 8.541: Sts 6.491 Se 6.67 Likewise, recri8talli8tion of the cis epimer gives a product which softens at 108 C and again at about 189
C (solvated), with 1, the following analysis: Analysis: (corrected for 4.95% water);
C: 50.27; H: 9.00 = Ni 6.05; S: 6.65.



   The trans epimer is about 2.2 times as active as lincomycin against S. lutea, about 2 times as well. active in the broth dilution assay, and 2.5 times as active in mice infected with S. aureus.



   The cis epimer is about 1/2 to 1/3 as active as the trans epimer, being roughly equal to lincomycin.

  <Desc / Clms Page number 40>

 
 EMI40.1
 



  11. Methyl 6,8-deoxy-6- (1-ethyl-4-utyl-, L-2-pyrrolidinecarboxamido) -1-thio-D-erythro "aD-qalacto-octopvranoside hydrochloride A mixture of 2, 0 gr of 6,8-deoxy-6 "(4-butyl-I) hydrochloride <-2-pyrrolidinecarboxamido) -1-thio-D-erythro-apDgalaato-oatoàyranoaide of methyl from part J, 1.5 ml of acetaldehyde, 150 mg of palladium at% on charcoal in 150 ml of methanol is shaken under hydrogen pressure of.



  35 pounds for five and a half hours. The catalyst is filtered off to give a residue consisting mainly of
 EMI40.2
 into the cis and trans epimers of methyl 6,8-deoXy-6- '(1-ethyl-4-butyl-L-2-pyrrolidinecarboxamido) El-thio-D-erythro-a-D-galacto-octopyranoside hydrochloride.

   L2. separation of epimers. . As described in part K2, the mixture of
 EMI40.3
 mothers of part Ll. (2 g) is chromatographidi on 200 g of silica gel using for elution a solvent system comprising ethyl acetate, acetone and water (8/4 / 1), Fractions 32 to 42 are, according to thin layer chromatography, the pure trans epimer and they are combined, while Fractions 49 to 64 are the essentially pure cis epimer and they are also combined. Fractions 43 to 48 are a mixture of the epimers which could be purified, by rechromography.

   Each epimer is dissolved in a few drops of dilute hydrochloric acid and the crystalline hydrochloride is precipitated by dilution with a large volume of ether.



   The crude trans epimer fraction @ 415 mg
 EMI40.4
 gives 340 mg (15.4%) of 6,8-deoxy-6- (trans-1-ethyl-4-butyl-L-2-pyrrolidinecarboxamido) -l-thio-D-drythro-aD-galacto- hydrochloride Crystalline methyl octopyranoside, melting point 144-151 C, Recrystallization from dilute acetone raises the melting point to 148-151 C.

  <Desc / Clms Page number 41>

 
 EMI41.1
 



  The 645 mg ci * epimer fraction gives 300 mg 1.1% I of â r 8-ddsoxy-â- o, s-1-ethyl-4-'buty3-L-2-pyrrolidineaarboxam hydrochloride (do ) Methyl -1-thio-D-eryùro-aD-galacto-oatopy-ranoside, crystalline, melting point 135-139 C.



  Recrystallization from dilute acetone gives crystals with a melting point of 134-138 C.



   The trans epimer isomer shows about 1 to 1.2
 EMI41.2
 times the activity of lincomycin in the test vis-à-vis "Si lutea, 2 to 4 times the activity of lincomycin vis-à-vis: organisms taking the gram, - and 8 times or more the activity of lincomycin against organisms not taking the gam.
 EMI41.3
 In mice, against S. aureus, trans-epimer is about twice as active as lincomycin. The cis epimer has about half the activity of the trans epimer.



   A separation of the cis and trans isomers is not a necessary phase since the 7-chloro derivatives of the mixed epimers are interesting in themselves. It is, however, desirable to maintain the content of the trans isomer at a high level since this is the most active form. By carrying out the process with this in mind, one can easily obtain mixed epimer products containing a ratio of trans epimer to cis epimer of 3/1 to 1/5.

   By substituting formaldehyde and acetaldehyde of parts K and L with other compounds oxode the formula R4RSCO, for example: propionaldehyde, acetone, butyraldehyde, isobutyl methyl ceone, benzaldehyde, phenylacetaldehyde, hydrocinnamaldehyde, acetophenone, propiophenone, butyrophyte
 EMI41.4
 none, 3-methyl-4-phdnyl-2-butanonée 2-methyl-5-phenyl-3-pentanone, 3-cyalopentanepropionaldehyde;

   'cyclohexanJbtaldehyde, cycloheptanecarboxaldehyde, 2,2-dimethylcycloprqp% aneacetaldehyde, 2,2-dimethylcyclopropyl methyl ketone, cyclopentyl methyl ketone, cyclobutyl methyl ketone, cyclobutanone, cyclohexanone, cyclohexanone and alx, cyclohexanone, and cyclohexyl or 4-methylcyclopropyl methyl ketone

  <Desc / Clms Page number 42>

 
 EMI42.1
 aralkyl 6,8-dideaoxy-6- (4-alkyl, 4-cycloalkyl-t or 4-aralkyl-L 2pyrrolidinecarboxamido) -1.thio-D-'rythro-an-; alacto-octopy. Appropriate ranoside, one obtains alkyl, cycloalkyl, and aralkyl 6, e-didsoxy-- (X RRCH - alky3 r 4-aycloalkyl and 4-aralkyl-L- ,.



  Z PYrroi id inecarboxamido -.- th.o-3-xythxo-t-D-galaato-octopyra nosides which, by treatment with thionyl chloride by the method of part A, give alkyl, cycloalkyl and aralkyl.
 EMI42.2
 



  '7-halo-G, 7r8-tridesaxy-6- (1-R4RCH-4-aikyi, 4-cycloalkyl-, and 4-aralkyl-L-2-pyrrolidinecarboxamido) -1-thio-a-D-galacto-octo-. corresponding pyranosides in which R4R5CH- estpropyl,
 EMI42.3
 isopropyl, butyl, and 4-methyl-2-pentylel benzyl, phenethyl, 3-phenylpropyl, 1-phenylethyl, 1-plienylpropyl, 1-phenylbutyl, 3-methyl-4-phenyl-2-butyl, and 2-methyl-5 -phenyl-3-pentyl, 3cyclopentylpropyl, 2-cyclohexylethyl, cycloheptyl-methyl, 2- (2,2-dimethylcyclopropyl) ethyl, 1- (2.2-dimethylcylopropyly ethyl, 1-cyclopentylethyl, 1-cyclobutylethyl, cyclobutyl, and cyclohexyl) 4-methylcyclohexyl.Using formaldehyde and acetaldehyde or other alkanals, for example propionaldehyde, butyraldehyde, valeraldehyde or aldehyde ca-
 EMI42.4
 protque with a 6,

   8-dideoxy-6- (4-aikyl-L-2-pyrrolidi, necarbôxamido) -1-thio-D-erythro-aD-galacto-octopyranoside in which the alkyl and the 4-alkyl are methyl, ethyl, propyl, butyl, pentyl, or hexyl, preferred starting compounds of the formula:
 EMI42.5
 

  <Desc / Clms Page number 43>

 in which X represents the hydroxy radical and R, HR1 and R3 consist of an alkyl of at most 6 carbon atoms, advantageously of 12 carbon atoms in the whole, Boni obtained, compounds which, by treatment with a reagent of Rydon, by the method of part A, give compounds of formula XXIV where X is halogen and R, HR1 and)

     are alkyl as defined above. The 5,8-dideoxy compounds of formula XXI. as characterized above, are active antibacterial agents comparable to the iinçomy-. cine. The 7-halo-6,7,8-trideoxy compounds of formula XXI, as characterized above, have the spectrum of antibacterial activity but they are significantly more active. Representative compounds of formula XXIV. where X is bromine, are given in the following tableRu.

  <Desc / Clms Page number 44>

 



   TABLE I Compound? of formula XXIV, where X is chlorine.
 EMI44.1
 
 <tb>
 <tb>



  R <SEP> HR1 <SEP> R3
 <tb> 4A <SEP> methyl <SEP> ' <SEP> Trans-ethyl <SEP> methyia <SEP> ' <SEP>
 <tb> 4B <SEP> methyl <SEP> cis-ethyl <SEP> methyl
 <tb> 4C <SEP> (7-bromo-7-deoxylincomycin <SEP> methyl <SEP> trans-propyl <SEP> methyl
 <tb> 4D <SEP> (7-bromo-7-deoxyallolincomycin) <SEP> methyl <SEP> cis-propyl <SEP> methyl
 <tb> 4E <SEP> (7-bromo-7-deoxylincomycin <SEP> E) <SEP> methyl <SEP> trans-propyl <SEP> ethyl
 <tb> 4F <SEP> (7-bromo-7-deoxyallolincomycin <SEP> E) <SEP> methyl <SEP> cis-propyl <SEP> ethyl
 <tb> 4G <SEP> ethyl <SEP> trans-propyl <SEP> methyl
 <tb> 4H <SEP> ethyl <SEP> cie-propyle <SEP> methyl
 <tb> 41 <SEP> methyl <SEP> trans-butyl <SEP>,

    <SEP> methyl
 <tb> 4J <SEP> methyl <SEP> cis-butyl <SEP> methyl
 <tb> 4K <SEP> methyl <SEP> trans-propyl <SEP> ethyl
 <tb> 4L <SEP> methyl <SEP> ' <SEP> cis-propyl <SEP> ethyl
 <tb> 4M <SEP> ethyl <SEP> trans-propyl <SEP> ethyl
 <tb> 4N <SEP> ethyl <SEP> cis-propyl <SEP> ethyl
 <tb> 40 <SEP> methyl <SEP> trans-butyl <SEP> ethyl
 <tb> 4P <SEP> methyl <SEP> cis-butyl <SEP> ethyl
 <tb> 4Q <SEP> methyl <SEP> trans-pentyl <SEP> methyl
 <tb> 4R <SEP> methyl <SEP> cis-pentyl <SEP> methyl
 <tb> 4S <SEP> ethyl <SEP> trans-butyl <SEP> ethyl
 <tb> 4T <SEP> ethyl <SEP> cis-butyl <SEP> ethyl
 <tb> 4U <SEP> methyl <SEP> trans-pentyl <SEP> ethyl.
 <tb>



  4V <SEP> methyl <SEP> cis-pentyl <SEP> ' <SEP> ethyl
 <tb> 4W <SEP> ethyl <SEP> trans-pentyl <SEP> methyl
 <tb> 4X <SEP> ethyl <SEP> cis-pentyl <SEP> methyl
 <tb> 4Y <SEP> methyl <SEP> trans-hexyl <SEP> methyl
 <tb> 4Z <SEP> methyl <SEP> cis-hexyl <SEP> methyl
 <tb> 4AA <SEP> butyl <SEP> trans-propyl <SEP> methyl
 <tb> 4AB <SEP> butyl <SEP> cis-propyl <SEP> methyl
 <tb> 4AC <SEP> ethyl <SEP> transpentyle <SEP> ethyl
 <tb> 4AD <SEP> ethyl <SEP> cis-pentyl <SEP> ethyl
 <tb> 4AE <SEP> butyl <SEP> trans-butyl <SEP> ethyl
 <tb> 4AF <SEP> butyl <SEP> cis-butyl <SEP> ethyl
 <tb> 4AG <SEP> butyl <SEP> trans-pentyl <SEP> methyl
 <tb> 4AH <SEP> -butyl <SEP>.

    <SEP> cis-pentyl <SEP> methyl
 <tb> 4Al <SEP> cyclohexyl <SEP> trans-propyl <SEP> methyl
 <tb> 4AJ <SEP> cyclohexyl <SEP> cis-propyl <SEP> methyl
 <tb> 4AK <SEP> butyl <SEP> trans-pentyl <SEP> ethyl
 <tb> 4AL <SEP> butyl <SEP> cis-pentyl <SEP> ethyl
 <tb> 4AM <SEP> pentyl <SEP> trans-pentyl <SEP> ethyl
 <tb> 4AN <SEP> pe <SEP> tyle <SEP> ois-pentyle <SEP> ethyl
 <tb>
 
Intermediates for the preparation of the above compounds corresponding to the preceding table where (1) X represents the hydroxy radical; (2) R3 represents hydrogen, (3) X represents the hydroxy radical and R3 represents hydrogen; (4) X is the hydroxy radical and R3 is the carbobenzoxy radical; (5) X represents bromine and R3 represents the carbobenzoxy radical;

   (6)

  <Desc / Clms Page number 45>

 X is the hydroxy radical, R3 is the carbobenzoxy radical and HR1 and H in the 4 position are replaced by the ylidene group, R1; (7) X represents bromine, R3 represents carbobenzoxy and HR1 and H in the 4 position are replaced by the ylidene group, R1; (8) X represents the hydroxy radical, R represents hydrogen and HR1 and H in the 4 position are replaced by the ylidene group, R1; (9) X is bromine, R3 is hydrogen and HR1 and H in the 4 position are replaced by the ylidene group, R1; (10) X is the hydroxy radical, and HR1 and H in the 4-position are replaced by the ylidene group, R1; and (11) X is bromine, and HR1 and H in position, 4 are replaced by the ylidene group, R1.



   When the lincomycin of part A is replaced by an alkyl, cycloalkyl or aralkyl 6-amino-6,8-dideoxy-1-thio-D-erythro-Ó-D-galacto-octopyranoside, the compounds of, the formula:
 EMI45.1
 in which R is: ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, doddcyl, trid6cyl ,. tetradecyl, pentadecyl. hexadecyl, heptadecyl, octadecyl, nonadecyl and eicosyl, or any of their isomeric forms, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cycloootyl, 2-methylcyclopentyl, 2,3-dimethylcy-clobutyl, 2-methyl and 3-cyclobutyl -cyclopentylpopyle:

     benzyl, phenethyl, 3-phenylpropyl, and Ó-naphthylmethyl, which compounds may be acylated by the methods of part I to form

  <Desc / Clms Page number 46>

 analogs of the corresponding 7-halo-7-deoxylincomycin, for example the compounds of formula XXIV given above, as well as the intermediates for these compounds.



  EXAMPLE 2
 EMI46.1
 "-chloro-7-deoxylincomycin D" 'T-chlora-6, 7, $ -trideoxy-6- (trana-4-propyl-L-2-pyrrolidinecaxboxamido) - 1-thio-L-threo-aD-galacto- methv octopyranoside
 EMI46.2
 
Chlorine is bubbled through a suspension of 11 g of triphenylphosphine in 200 ml of acetonitrile until a binding of 2.84 g is noted, the reaction mixture is cooled to 26 'and added. 1.72 gr of N-demethyllincomycin hydrochloride (lincomycin D hydrochloride). The solution is stirred at room temperature for 18 hours. After addition of 10 ml of ethanol, the solvent is distilled off in vacuo.



  The residue is stirred with 100 ml of ethyl acetate-ether (1/1).



  The mixture is filtered and the residue is partitioned between 15 ml of ethyl acetate and 15 ml of ae. Ethyl acetate is washed once; with 10 ml of water and the combined aqueous extracts are re-exhausted with ethyl acetate. The aqueous solution gives a residue of 3.05 g on lyophilization. Caolide is dissolved in methanol, 10 g of silica gel are added and the solvent is evaporated off to obtain a free-flowing powder.

   This powder is placed at the top of a chromatographic column of 100 g of silica gel and this column is eluted with chloroform-methanol (4/1) The fractions containing the product, as determined by

  <Desc / Clms Page number 47>

 thin layer chromatography, are combined and evaporated to give a crude material weighing 1.27 g. This solid is dissolved in acetone and acidified with hydrochloric acid
 EMI47.1
 diluted. 7-chlOrO-7-deOXY-N-methYllificin hydrochloride. with a melting point of 212-216 C weighing 0.73 gr, settles on cooling.

   Recrystallization from acetone-water gives 550 mg of hydrochloride, melting point 217-221 ° C (decomposition).



  Analysis:
 EMI47.2
 Calculated for C17H32C12NOSS.HCl 1 Cs 45.62j Hs 7.2 1 Ni 6.26 Found 8 Cs 45.891 Bi 7 # 661 Ni 6.55 7-a # loro-7-deoxy-N-demethyilinaomycin hydrochloride is about 8 times as active as lincomycin viz-àvia gram-taking bacteria and it is at least 8 liver too
 EMI47.3
 active as linaomyaine against bacteria. not taking the gram..It is about twice as active as 7 x 7-chloro-7-dessoxylincomycin. This is unexpected because N-desmethyl1incomycin is less active than lincomycin.



   By substituting brotae for chlorine, 7-bromo-7-deoxy-N-demethyllincomycin is obtained. Instead of triphenylphosphine, triphenyl phosphite can be used. Further, in this case, methyl halide can be used instead of halogen.



   Lincomycin D can be prepared as described in U.S. Patent No. 3,329,568., .. ''

  <Desc / Clms Page number 48>

 EXAMPLE 3
 EMI48.1
 7-ehloro-6., 8-trideaaxy-6- (tranA- and aia-4-penyl L-2-pyrrolidinecarboxam ± do) -1-thio-L-threo-a-D-qalactopctopyranoside e mµlbyle
 EMI48.2
 Has a colorless solution of triphenyl-dichloride
 EMI48.3
 phosphine, prepared treated.

   22 gr of triphenylphosphine in 400 ml of acetonitrile with Se $ 8 gr of chlorine, 4 gr is added
 EMI48.4
 6 # 8-diddnoxy-6- (trans- and oifl-4-pontyl-L-2pyrrolidinecarboxamido) -1-thio-b-erythro-ab-gelacto-octopyranoside hydrochloride (mixture of cis and trans isomers , having a melting point of 212-214 ° C). After stirring at 26 ° C. for 18 hours, 15 ml of methanol are added and the empty mud solution is distilled off. The residue is shaken with 250 ml of ethyl acetate-ether (1/1) and filtered. The residue ,. namely 13.7 g, is distributed between water and ethyl acetate and the product is recovered from the aqueous solution by lyophilization.

   The freeze-dried product, namely 8.5 g, is further purified by chromatography on silica gel using chloroform-methanol (4/1) for the elution. A large fraction of 2.09 gr is dissolved in acetone and acidified with hydrochloric acid, then filtered and dried to obtain 1.15 gr (27.6%) of 7-chloro-6.7 , 8-trideoxy-6- (trans- and cis-4-pentyl-L-2-pyrrolidi- necarboxamido) -l-thio-L-threo-aD-galacto-octopyranoside of me-
 EMI48.5
 thyle.

   Melting point s222-223 ° C (decomposition) f 7D. +1398 (H20)

  <Desc / Clms Page number 49>

   Analysis
 EMI49.1
 Calculated for e xio 8 8 CI 47.99j And 7.631 Si 5.891 Si â.'4 Found: (correction for 0.58% water):
 EMI49.2
 Ci'47.851 And 7.841 Ni 6.01 gs 6.01
The mixture of cis and trans isomers, used as a starting material in this example, was prepared as follows
 EMI49.3
 B. 4-pentylidene-1-carbobenzôxy-L-proline and its only dicyèlohexylamine
By following the procedure of Part F of Example 1, replacing butyltriphenylphosphonium bromide with pentyltriphenylphosphonium bromide, 4-pentylidene-1-carbobenzoxy-L-proline and its dicyclohexylamine salt are obtained.

   The free acid is an oil but the dicyclohexamine salt, after recrystallization from acetonitrile, has a melting point of 124-J
 EMI49.4
 128 * C, 7D optical rotation. -6 * (and 0.762 CHCl3), and the following analysis Calculated for C30H46N2O4 Ci 72.251 Hi 9 # 301 Nz 5.62 Found s Cs 72.3SI. Hi 9.521 Ns 5.97 C. 4-pentyl-1-carbobenzoxy-L-proline
Hydrogenation of the oily free acid of Part B, by the method of Part D of Example 1, gives 4-pentyl-1-carbobenzoxy-L-proline as an oil.
 EMI49.5
 



  D. 6,8-dideaoXy-6- (4-pontyl-1-carbobeÀzoxy-L-2-pyrrolidinecarboxami4e) -1-thio-D-6rythro-a-D-galaotooctaovranoide methyl arllW r111-wwnr'1
Following the procedure of Part 1 of Example 1, substituting 4-butyl-1-carbobenzoxy-L-proline with 4-pen-
 EMI49.6
 .tyl-1-carbobenzoxy-L-, roline (the oil from Part C), we obtain, '6,8-dideaxy-6- (4-pentyl-1-carbobenzoxy - 2-pyrrolid3necarboxamido) -1 Methyl thio-D-erythro-aD-galacto-octopyraÀo * ide, having a melting point of 191-193 C, optical rotation

  <Desc / Clms Page number 50>

 
 EMI50.1
 çt +108 e (c 0.722, r blet3H) and the following analysis at Calculated for C27 H42v 20a0 at Ci 58.46t Us 7.631 Ut 5.05 Found Ce 38.321 Ht 7.321 Ni 4.95 1.

   6,8-did '@ oxy-6- (4-p * ntyl to-1µ1 pyrrol id inecarboxamido) -1-th io-D-6rythro-C-D hydrochloride ...;' Methyl qalacto-octopyranoside.



  Hydrogenolysis of d, 8-didësoxy-6- (1-rarbobenzocy 4-pentyl-L-2-pyrolidinecarboxamido) -l-thio-D-drythro- -D-galacto.d octopyranoside by the method of Part J of l Example 1 gives methyl 6,8-dideoxy-6- (4-pentyl-L-2-pyrrolidinecrw boxamido) -1-thio-D-erythro-aD-galacto-octopyranoside hydrochloride, having a melting point of 212-214 C, optical rotation
 EMI50.2
 % / to7- of +141 '(e = 0.968 = H2O), and the following analysis s --Calculated for C19H37N2O6SCl 1.



   C: 49.931 H: 8.161 N: 6.131 S: 7.02 Found: (correction for 5.43% H2O)
 EMI50.3
 ci 50.22; Ha 7.961 Na 6.09 Ss 7.18
Instead of chlorine in previous Examples 2 and 3, bromine can be used with formation of the corresponding 7-bromo analogs. In addition, instead of lincomycin D,
 EMI50.4
 N-demdthyl-3-carbobonzoxylincomycin and its analogs prepared as described above can be used, the following example is purely illustrative. EXAMPLE 4
 EMI50.5
 Part A. N-d'm'th1.M9arbcbenICKYlincomvcin.



  We repeat the preceded of Example 1, Party. ' . X, using propyltriphenylph08pbium bromide instead of butyltriphenylphosphonium bromide. The resulting mixture of cis and trans isomers is chromatographed by the method of Part K2 to give the trans isomer, namely N-demethyl-N-carbobenzoxylincomycin, with a melting point of 176-178 C: [Ó ] D = +109, and the following analysis:

   

  <Desc / Clms Page number 51>

 
 EMI51.1
 Calculated for C25H3SK20eS s Ci 57.071 Et 7 # 271 Nit 5.23 Found t Ci 56.84t Ri 7.28t Si 5.37 Part B.? -Chloro-'9-deoxy-N-clmethyl-N-carbtbenzoxvlinoomvoine j Add 1 g of 3-demethyl-N-carbobentoxylindotaycine to a solution of triphenylphosphine dichloride prepared by adding 710 mgr of chlorine to 2.88 g of triphenylphosphine in 25 ml of acetonitrile. After 2 hours, 10 ml of methanol are added and the solvent is evaporated off. The residue is chromatographed on 100 g of silica gel using chloroform-methanol (4/1) for the elution. We obtain a fraction of 717 mgr, which is a mixture
 EMI51.2
 of 7-chloro-7-dôsoxy-N-demethyl-N-carbobenzoxyiincotaycine and triphenylphoophine oxide.

   This mixture is used directly in the next Part.
Part C. 7-Chloro-7-deoxy-N- hydrochloride
 EMI51.3
 demethyllincomycine 7-chloro-7-d6soxy-N is dissolved. arbobenzoxy - crude lincomycin from the previous part (717 mgr) in 25 ml of methanol and 250 mgr of a catalyst at% palladium on carbon is added. The mixture is shaken under 40 pounds of hydrogen pressure for 2 hours. The catalyst is filtered off and the solvent is distilled off in vacuo. The residue is purified by chromatography on silica gel (100 g of silica gel) methanol-chloroform si / 4 for elution) to obtain 103 mgr of 7-chloro-7-deoxy-N-d6methyl-lincomycin hydrochloride. . This product is dissolved in acetone and acidified with dilute hydrochloric acid.

   Hydrochloride is obtained
 EMI51.4
 of crude 7-chloro-7-denoxy-N-demethyllincomycin, melting point at 227-229 C and weighing 95 mgr. A standard curve (test against S-lutea) indicates antibacterial activity 1.7 times that
 EMI51.5
 7-chloro? -deoxylincomycin.

  <Desc / Clms Page number 52>

 
 EMI52.1
 



  Instead of separating the iaornerea oit and trans, the mixture can be treated by the method of Parts B and C to obtain N- (1-carbobenzoxy-4-trans- and ais-propyl-lî-prolyl) -7- methyl M- (4-tran @ - et ciepropyl-L-prolyl} -7-chloro-7-d.oxy-c-thiolinco.aminide, methyl ehloro-7desoxy-ct-thiolinco.aminide, analogues 8- and 4-corresponding and the corresponding 7-bromo analogs can be prepared in a similar fashion. Or, a methyl 7-halo-7-deoxy-α-thiolincolaminide or an analogue thereof can be acylated with a cis and trans-1-carbobenzoxy-4-alkyl-, 4-cycloalkyl-, or 4-aralkyl-L-proline The following example is merely illustrative.



  EXAMPLE 5
 EMI52.2
 Part A. Methyl N- (1-earbobenxoxy-4-trans- et.cis-propyl-prolyl) -7-ahloro-7-dedoxy-lincoaaminide
 EMI52.3
 
 EMI52.4
 Ala and trans-1-carbobonzoxy-4-propyl are dissolved! L-proline (2.33 g) in 150 ml of acdtonitrile containing 1.12 ml of triethylamine. The solution is cooled to 0 and 1.18 ml of iaobutchloroformate is added. After 10 minutes at 0, a solution of 2 ', 17 g of methyl 7-chloro-7-deaoxy-a-thiolincoaaminide in 40 ml of actonitrile and 40 ml of water is added. The mixture is stirred for 2 hours at room temperature and the solvent is distilled off in vacuo to obtain a crystalline residue.



   The crystals are collected by filtration, washed

  <Desc / Clms Page number 53>

 
 EMI53.1
 and dried. The production of methyl Nd-carbobenzoxy-4-trans and cis-propyl-L-prolyl) -7-chloro-7-deaoxylincomsaminide, melting point 180-183 C, is 3 , 36 gr. A portion is recriitallineed several times in ethanol.

   This portion melts at 189-192 C; Analysis: Calculated for C25H37ClN2O7S
 EMI53.2
 Ci 55 # 081 Hs 6 # 841 Cli 6.51, Nt 5.14 Found s Ci 54 # aol Hs 7.151 Cls 6 # 591 Nt 5.16 Part B, N- (4-trana and cia-propyl-L hydrochloride - 'prolylj-7chloro-7-déaoxy, methvie linaosaminida - ######. #########; #####
 EMI53.3
 
A portion of the above crude product is dissolved in 50 ml of methanol and 0.5 g of a 10% palladium-on-charcoal catalyst is added. The mixture is shaken under a pressure of 35 pounds of hydrogen for 4 hours. . Thin layer chromatography shows partial hydrogenolysis. An additional O. S g of catalyst is added, and the hydrogenation is continued for 18 hours. The catalyst is separated by filtration.

   The residue is chromatographed on silica gel and the more polar fraction * is collected. It weighs 185 mgr and is converted into the hydrochloride in the usual way, which gives 150 mgr of crystals, with a melting point of 220-222 * C (decomposition), with a power equal to about 80 times that of lincomycin.



   .

  <Desc / Clms Page number 54>

 



   Another portion of the crude product (2.9 g) is dissolved in 500 ml of methanol and 6.0 g of a 10% palladium on carbon catalyst is added. The mixture was shaken under 35 pounds of hydrogen pressure for 18 hours. The catalyst is filtered off and the clear liquid phase is evaporated off. The crude solid product is converted to the hydrochloride. Crystallization of the crude product from acetone-water gives 15.08 g of crystals, melting point 218-223 ° C (decomposition). Recrystallization from water gives an analytical sample, with a dB melting point of 228-234 Ci [Ó] D = +159.
 EMI54.1
 



  Calculated for Cl'Ii3058C12 i Ci 45.631 fis i, 21 Ni 6.26 Found: (correction for 3.91% H20)
C: 45.851 H: 7.51; N: 5.86
 EMI54.2
 Part C. Methyl 7-chloro-7-deoxy-a-thiolinao @ aminide is prepared as follows.
 EMI54.3
 
 EMI54.4
 To a suspension of 197.2 g of triphenylphoaphine in 1.5 liters of anhydrous acetonitrile, 52.5 g of chlorine are added. With stirring, 18.75 grams of methyl a-thiolincosaminide (U.S. Patent No. 3,179,565) are added. After two and a half hours at room temperature, 50 ml of methanol are added. The mixture is concentrated into a thick syrup. This concentrate is diluted with methylene chloride and extracted three times with water.

   The aqueous extracts are washed twice with methylene chloride,

  <Desc / Clms Page number 55>

 The extracts are made alkaline with caustic soda and extracted repeatedly with methylene chloride. The organic extract is dried and evaporated in vacuo. The residue is chromatographed on 1.1 kg of silica gel using chloroform.
 EMI55.1
 methanol (4/1) potir: 3 'dilution, The main fraction chosen on the basis of its profile in thin layer chromatography weighs 4.4 gr. Recrystallization from methanol-water gives 2.73 gr '
 EMI55.2
 of methyl 7-chloro-7-deoxy-a-thiolincosaminide, mp 178-181, Analysis! : Calculated for C9H18ClNO4S.



   C: 39.77 = H: 6.67 = N: 5.16 = Si 11.801 Cl: 13.05
 EMI55.3
 Found s Cs 39.91: Hs 7.021 Nt 5.57 = Si 11.99 =, Cls 13.35
Part A cis and trans-1-carbobenzoxy-4-propyl-L-proline is prepared as follows.
 EMI55.4
 



  Part D. 1-carbobenzoicy-4-propylidèr, s-L-psoline and its dicvclohexvlamine salt
3.8 g of sodium hydride are heated with 75 ml of dimethylsulfoxide at a temperature of 70-75 C until the reaction is complete. After cooling to 20 ° C., 30.8 g of propyltriphenylphosphonium bromide is added and the resulting red solution is stirred for 30 minutes to ensure.
 EMI55.5
 total reaction. A solution of 5.2 g of 4-cdtc-1-aarbobenzoxy-L-proin in 15 ml of dimethylsulfoxide is added over a period of 15 minutes and the resulting mixture is stirred for 20 minutes at 26 C, and then at 70 C for 4 hours.

   The reaction mixture is cooled, 100 ml of a 5% aqueous solution of potassium bicarbonate and 100 ml of water are added, and filtered.



  The filtrate was washed twice with 150 ml portions of ether and the ether was recipitated after stripping again with bicarbonate. The solutions are combined with bicarbonate, diluted with 200 ml of water and acidified with 4N hydrochloric acid.

  <Desc / Clms Page number 56>

 



  The acidified mixture is extracted with 3 portions of 200 ml of ether, The combined ethereal extracts are washed with three portions / of 50 ml of a saturated aqueous solution of sodium bisulfite, then with water, and finally on. dries over anhydrous sodium sulphate, Evaporation of the solvent gives 5.7 g of a residue
 EMI56.1
 solid which is 1-carbobenzoxy-4-propylidne-L-proline. This residue is dissolved in 18 ml of acetonitrile and
 EMI56.2
 treated with 2.8 ml of dicyclohexylamine. dicyelohexyl- e ael, crystalline amine, 5.2 gr (55% yield), melts at 154-157 C.



  After three recrystallizations from acetonitrile an analytical sample is obtained which melts at 164-166 ° C and which has a total
 EMI56.3
 7D tion of -8 (c - Ot3898 gr / 100 ml; CHC13>.



    Analysis: Calculated for C28H42N2O4 * Ci 71.45; H: 9.00: N: 5.95.



   Found C: 71.77; H: 9.39; N: 5.1
 EMI56.4
 Part E. cls and trana.-1-carbobenzoxy-4-propy1-L-proline
10 g of the amino salt from Part D are shaken with ether and 2% caustic potash. The aqueous layer is separated and acidified. Extraction with methylene chloride leads to the isolation of 6 g of an oily acid. A mixture of 2 g of this oily acid and 800 mgr of a 7% platinum catalyst on Dowex-1 in 50 ml of methanol was shaken under 40 pounds of hydrogen pressure for 17 hours.

   The catalyst is filtered off and the solvent is distilled off in vacuo, leaving a residue of 2 g of an oil.
 EMI56.5
 Thin layer spelling, using ayatemethanol-5% ammonium hydroxide and a permanganate-periodate indicator indicates that the double bond is hydrogenated. Ninhydrin gives a negative test.

   This product is resistant to crystallization and is used without purification in Part A.

  <Desc / Clms Page number 57>

 EXAMPLE 6 '
 EMI57.1
 Methyl 7-chloro-6,7 # 8-triddnoxy-6- (cis- and trana-4-hexyl-pyrrolidineaarboxamido) -1-thio-L-hreo-a-D-galâcto-octouyranoide
Following the process of Example 5, substituting the propyltriphenylphosphonium bromide of Part D with hexyltriphenylphosphonium bromide, the dicyclohexylamine salt of N-carbobenzoxy-4-hexylidene-L-proline, d 'is obtained. a point
 EMI57.2
 of fusion of 115-119 "Cl the N-carbobenzoxe-4-cis and trans-hdxylL-proline form an oil;

   7-ohloro-6,7,8-trideoxy- 6- (1-carbobenzoxy-4-cie- and trans-hexyl-L-2-pyrrolidinecarboxamido) -1-thio-L-threo-aD-galacto- methyl octopyranoside, 4'a; :, mp 185-187 Ci and 7-chloro-6,?, 8-triceoxy-8 = (4-cie and trans-hexyl-L-2-pyrrolidipe-carboxapido ) Methyl -1-thio-L- 'threo-aD-galacto-octopyranoide, melting point 212-215 C (decomposition). 'EXAMPLE 7
By following the process of Example 5;

   replacing
 EMI57.3
 methyl α-thiolincoaaninide with methyl epi-a-thioüneosaminide (6-amino-6,8-dideoxy-1 thio-L-threo-aD-galacto-octopyronoide of methyl) ', one obtains the 7-chloro-7- deoxy-epilmcoNycin, D (7-chloro-6,7,8-trideoxy-6- (trans-4-propyl-L-2-pyrrolidineçair- boxamido) -l-thio-L-threo-aD-galacto-octopyranoside methyl) and its cis isomer.



   Methyl epi-a-thiolincosaminide is obtained by hydrazinolysis by the method of U.S. Patent 3,179,595 epilincomycin which is prepared as follows.

  <Desc / Clms Page number 58>

 
 EMI58.1
 



  A. .4-0-i89propvlidne l1noomvoine
 EMI58.2
 
A solution of 9.8 gr of lincomycin in 150 ml of acetone is added to a solution of 9.8 gr of p-toluenesulphonic acid monohydrate in 100 ml of acetone with good agitation and avoiding exposure to moisture. The mixture is stirred at room temperature for 1 hour, after which 100 ml of anhydrous ether is added and stirring is continued in an ice bath for 1/2 hour. The mixture is filtered and the solid is dried in vacuo. at 50 * Ci the production is 13.35 gr (85.5%) of
 EMI58.3
 3,4-o-isopropylidwnelincomycin p-tolueneaulfonate.

   An additional 1.15 gr (7.4%) can be recovered from the mother liquors by adding 350 ml of anhydrous ether to the mother liquor from the previous filtration operation, and then

  <Desc / Clms Page number 59>

 cooling the solution for 1 hour. The 14.5 g thus obtained are suspended in 200 ml of ether and vigorously shaken with 125 ml of a 5% solution of potassium bicarbonate.



    ,.



  The aqueous layer is again exhausted with 2 portions of 100 ml of ether.



  The ethereal extracts are washed with 50 ml of saturated sodium chloride solution and then filtered through anhydrous sodium sulfate. The ether is evaporated in vacuo, leaving
 EMI59.1
 7.9 gr (73.1%) of 3, 4-O-isopropylidénelinaomycin which is dissolved in 25 ml of ethyl acetate and concentrated to) approximately 10 to 15 ml, The concentrate is left at the rppos at the room temperature for several hours and then refrigerated overnight.

   The crystals are filtered from solution and washed lightly with cold ethyl acetate, the yield is 4.55%.
 EMI59.2
 gr (42.2%) of 3,40-isopropylideneincomycin having a melting point of 126-128 ° C and an optical rotation of [Ó] D of 101-102 (c = 1 methylene chloride). ,
 EMI59.3
 8. - h o- - - o om A a solution of 6 gr (0.0135 mol) of isopropylidene-. lincomycin in 75 ml of pyridine, 12 g (excess) of chromic oxide are added. The solution heats up to about 20 C. After 1 hour, the mixture is added to a solution containing 250 ml of ethyl ether and 250 ml of ethyl acetate. Then filtered and evaporated until a syrup (8.4 g) is obtained.

   This syrup is distributed in a countercurrent distribution at 500 trans-
 EMI59.4
 ferts using the water-ethyl acetate-ethanol-cyclohexane (1/1/1/1) system. 7-Dehydro-3,4-0-isopropylidene-lincomycin was isolated as peak fraction from tubes 330 - 380, K - 2.45. '
 EMI59.5
 Calc analysis for s C31B36N2O6S C: 56.72; H: 8.161 Na 6.30; S: 7.21
 EMI59.6
 Found, 1 Cs 56.37t and 7.62e Na 6.51; Sa 6.84 ,. '

  <Desc / Clms Page number 60>

 
 EMI60.1
 G "3.4-0.-iaaproaylidene-epilinconrcin To 1.6 g of pure 7-dehydro-3,4-0-isapropylidènelincomycin (Craig) in 75 ml of methanol, 400 mg of sodium borohydride are added.

   After 1.5 hours, this solution is evaporated to dryness on a rotary evaporator. The residue is added to 25 ml of water and 3 livers are extracted with 25 ml of methylene chloride in each case. The extract is washed back with 15 ml of water, and dried over magnesium chloride and evaporated to dryness. The residue (1.4 g) is distributed in a dis-
 EMI60.2
 tribution yy against the current at 500 transfers, using the solvent system consisting of water, ethyl acetate, ethanol and cyclohexane (1/1/1/1). a single point which corresponds to the theory to the value K = 1.05.



  The material in tubes 240 to 280 is isolated as alum syrup, Analysis Calculated for C21H38N2O6S
 EMI60.3
 Cs 6.4'i1 lis 8.581 lasts d, 3'1 Si 7.18 Found! Ci 56.24 Hi 8.54t Ni 6.131 Si 7.01 A thin layer chromatography shows that this
 EMI60.4
 matter consists of two substances. One is 3,4-0-iaoprpy lidënelincomycin, while the other is 3, 4-0-, ropropyl3din. slightly slower moving dpilincomycin
D.

   Epilincomycin
The syrup from part C is stored at room temperature for 5 hours in a solution containing 60 ml of 0.25N hydrochloric acid and 40 ml of ethanol. It is then maintained at 0 C for 4 days. After neutralization with sodium bicarbonate, evaporated to 25 ml, then extracted with chloroform. The extract is washed with a little water and dried over magnesium sulfate, then evaporated to a residue.



  Thin layer chromatography of the residue shows that there is
 EMI60.5
 two substances, both of which are active against S. lutea,

  <Desc / Clms Page number 61>

 The residue is chromatographed on a 14 inch x 3/4 inch Florisil column (synthetic silicate of the type described in US Pat. No. 2 (393,625), this column being gradient eluted with a continuously varying solvent deputing 100% Skellysolve. B (technical hexane) up to 100% acetone.



  The total volume is 5000 ml. The following two compounds are thus separated.



  Fraction 1: Tubes 53-65 (40 ml portions) epilincomycin
Titration: 450 mcg / ml /.



  Analysis calculated for C18H34N2O6S:
Ci 50.921 H: 8.551 Ni '6.601 S: 7.56
Found: Ci 50.191 H: 7.911 Ni 6.05; Si 6.42 Fraction II: Tubes 73-104. Lincomycin
Titration: 950 mcg / mg ...



    EXAMPLE 8 '. 7-bromo-7-deoxyepilincomycin
The process of Example 1, substituting epilincomycin for lincomycin, gives 7-bromo-7-deoxyepilincomycin of the formula
 EMI61.1
 Bout forms the free stock * and forms hydrochloride.



   By substituting the linoomycin of this example with the lincomycin analogs * of formula II. In which Z, R, R1, R2, R3 of the group Ac are as given previously, we

  <Desc / Clms Page number 62>

 obtains the corresponding 7-halo-7-deoxyepilincomycin analogs of the formula
 EMI62.1
 in which Z, R, R1, R and R3 of the group Ac are as given previously. All the compounds which have been described above therefore have their counterpart in the opposite configuration, ie a configuration derived from the 7-epi form.



  If an inversion is achieved by substituting the 7-hydroxy radical with a Rydon reagent, then epi compounds which have the L-threo configuration are converted to the D-erythro configuration.



  In either case, both D-erythro and L-threo forms are obtained, depending on whether normal lincomycins (D-erythro) or epilincomycins (L-threo) are used.



   Although the methods of the invention have been described with reference to the preparation of particular compounds, it will be understood that the method is. broadly applicable to compounds of formula II in which Ac and R are radicals which do not react with Rydon's reagent. Thus, in the general process according to the invention, Ac can be hydrogen or any acyl which is not reactive with the Rydon reagent, and R can be any alkyl, whatever either the number of carbon atoms, or any other radical, eg aralkyl such as benzyl, naphthylmethyl and benzydryl, or similar hydrocarbon radicals which do not react with Rydon's reagent ...

 

Claims (1)

CLAIMS 1. A compound of the formula: EMI63.1 wherein R is alkyl of 20 carbon atoms at most and Ac is the acyl radical of a 4-substituted L-2-pyrrolidinecarboyxlic acid having the formula selected from the group. including: EMI63.2 formulate in which R1 and R2 are selected from the group consisting of alkylidenes of 20 carbon atoms at most; cycloalkylidenes of 3 to 8 carbon atoms at most and araylkylidenes of at most 12 carbon atoms, and R3 is selected from , the group comprising hydrogen and HR2, 2. A compound of the formula: EMI63.3 <Desc / Clms Page number 64> in which R, OR 1 and HR2 are constituted by an alkyl, lower, 3. A compound according to claim 2, wherein R is methyl, HR1 is propyl and HR2 is methyl, 4. A compound according to claim 2, wherein R is methyl, HR1 is propyl and HR2 is hydrogen, 5. A compound according to claim 2, wherein R is methyl, HR1 is propyl and HR2 is ethyl. 6. A compound according to claim 2, wherein R is ethyl, HR1 is propyl and HR2 is methyl. 7. A compound according to claim 2, wherein R is ethyl, HR1 is propyl, and HR2 is hydrogen. 8. A compound according to claim 2, wherein. R is ethyl, HR1 is propyl and HR2 is ethyl. 9. A compound according to claim 2, wherein R is propyl, HR1 is propyl and HR2 is methyl. 10. A compound according to claim 2, wherein R is propyl, HR1 is propyl and HR2 is hydrogen. 11. A compound according to claim 2, wherein. R is propyl, HR1 is propyl and HR2 is ethyl. 12. A compound according to claim 2, wherein R is butyl, HR1 is propyl and HR2 is methyl. 13. A compound according to claim 2, wherein R is butyl, HR1 is propyl and HR2 is hydrogen. 14. A compound according to claim 2 wherein R is butyl, HR1 is propyl and HR2 is ethyl. 15. A living compound of claim 2, wherein R is methyl, HR1 is pentyl and HR3 is methyl. 16. A compound according to claim 2, in which: R is methyl, HR1 is pentyl and HR2 is hydrogen. 17. A compound according to claim 2, wherein R is methyl, HR1 is pentyl and HR2 is ethyl. 18. A compound according to claim 2, wherein R is methyl, HR1 is butyl and HR2 is methyl. <Desc / Clms Page number 65> 19. A compound according to claim 2, wherein R is methyl, HR1 is butyl and HR2 is hydrogen. 20. A compound according to claim 2, wherein R is methyl, HR1 is butyl and HR2 is ethyl. 21. A compound according to claim 2, wherein R is ethyl, HR1 is butyl and HR2 is methyl. 22. A compound according to claim 2, wherein R is ethyl; HR1 is butyl - and HR2 is hydrogen. 23. A compound according to claim 2, wherein R is ethyl, HR1 is butyl and HR2 is ethyl. 24. A compound according to claim 2, wherein R is propyl, HR1 is butyl and HR2 is methyl. 25. A compound according to claim 2 wherein. R is propyl HR1 is butyl and HR2 is hydrogen. 26. A compound according to claim 2, wherein R is propyl, HR1 is butyl and HR2 is ethyl. 27. A compound according to claim 2, wherein R is butyl, HR1 is butyl and HR2 is methyl. . 28. A compound according to claim 2, wherein R is butyl, HR1 is butyl and HR2 is hydrogen. , 29. A compound according to claim 2 wherein R is butyl, HR1 is butyl and HR2 is ethyl. 30. A compound according to claim 2, wherein R is cyclohexyl, HR1 is butyl and HR2 is methyl. 31. A compound according to claim 2, wherein R is cyclohexyl, HR1 is butyl and HR2 is hydrogen. 32. A compound according to claim 2, wherein R is cyclohexyl, HR1 is butyl and HR2 is ethyl. , 33. 7-Bromo-7-deoxylincomycin. 34. The process for preparing the compounds of the formula: <Desc / Clms Page number 66> EMI66.1 in which R and Ac are as defined above, which comprises the reaction phase of a compound of the formula: EMI66.2 with a Rydon reagent & a temperature effective to replace the 7-hydroxy group by * a halogen, R and Ac being radicals which are not reactive with the Rydon reagents, 35. A process according to claim 34 wherein R is alkyl and Ac is acyl unreactive with Rydon's reagent 36. A process according to claim 34 wherein R is alkyl of not more than 20 carbon atoms and Ac is the acyl radical of a 4-substituted L-2-pyrrolidinecarboxylic acid having the formula selected from the group consisting of: EMI66.3 <Desc / Clms Page number 67> formulas in which R1 and R2 are chosen from the group comprising alkylidenes of 20 carbon atoms at most, cycloalkylidenes of 3 to 8 carbon atoms at most and aralky-. lidenes of 12 carbon atoms or less, and R is selected from the group consisting of hydrogen and HR2. 37. A process for the preparation of 7-halo-7-deoxylincomycins D and their analogs and isomers which comprises acylation of a compound of the formula: EMI67.1 with a compound of formula 1 EMI67.2 for format a compound of the formula: EMI67.3 formulas in which R 1 and R 1 represent an alkyl of 20 carbon atoms at the most, a cycloalkyl of 3 to 8 carbon atoms. at most bone or aralkyl of at most 12 carbon atoms, Z is a separable protecting group, and X is chlorine or bromine; and then separating the group Z to form a compound of formula <Desc / Clms Page number 68> EMI68.1 in which R1, R2 and X are as defined * previously, 38. A process for the preparation of lincomycin D and its analogs and isomers, which comprises reacting a compound ' EMI68.2 of the formula 1 -,? 3 ,, / H / N CH3 'Q HO 1 Off in which R and R 1 represent alkyl of 20 atomic atoms. of carbon at most, cycloalkyl of 3 to 8 carbon atoms at most or aralkyl of at most 12 carbon atoms, and Z is a separable protecting group, with a chlorinating or brominating reagent of Rydon at a temperature effective to replace the 7hydroxy group with chlorine or bromine, and then the separation of the Z group to form a count of the formula: EMI68.3 <Desc / Clms Page number 69> in which a and R1 are as defined above, and X represents chlorine or bromine. 39. Process for preparing a compound of the formula EMI69.1 wherein R and HR1 are alkyl of at most 20 carbon atoms, cycloalkyl of 3 to 8 carbon atoms at most, or aralkyl of at most 12 carbon atoms, halo represents chlorine or bromine, and R3 represents hydrogen, this process comprising the N-acylation of a compound of the formula: ' EMI69.2 in which R and halo are as defined above, with an acyl group of the formula: EMI69.3 <Desc / Clms Page number 70> in which R1 and R3 have the meaning given .. EMI70.1 40, The 7-halo-7-de.oxylinoomyoin .. and the 7-hAlo. EMI70.2 7-ddsoxydpilincomycins. 41. The new compounds of the formula EMI70.3 wherein R and R1 are alkyls of at most 20 carbon atoms, cycloalkyl of 3 to 8 carbon atoms at most, or aralkyl of at most 12 carbon atoms, and X represents chlorine or bromine. 42. The new compounds of the 'formula .. EMI70.4 where R and HR1 are alkyls of 20 carbon atoms EMI70.5 at most, csyolaalkyls of 3 to 8 carbon atoms at most or aralkyls of at most 12 carbon atoms, halo represents EMI70.6 chlorine or bromine, and R3 represents hydrogane, a hydrocarbyloxycarbonylop trityl d1phenyl- (p-.methoxyphenyl) methyl, bis '' (p-m6thoxyphenyl) -ph6nylm6thylet benzyl or nitro-benzyl radical. <Desc / Clms Page number 71> 43. New compounds, their intermediates, their analogues and isomers, as well as their methods of preparation. as described above, in particular in the examples, given ..