DE1620607A1 - Process for the preparation of lincomycin and analogous compounds - Google Patents

Description

Dr. Expl.

Dr. Walter Beil Alfred Hoeppener '

Dr. Ham Joachim Wolff I2.ftu9.1965

Dr. Hans Chr. Beil

Lawyers' Frankfurt a. M.-Höchst

ΤΛ3126 «

Our no.11859

The Upjohn Company Kalamazoo (Michigan, USA)

Process for the production of Lincomycin and analog connections.

The present invention relates to a method for the preparation of lincomycin and analogous isomeric compounds thereof.

Lincomycin is an antibiotic obtained as the metabolic product of a lincomycin-producing aetinomycete. A process for the preparation, isolation and purification of lincomycin are% n * U S. Patent 3,066,912 described. In connection with the present invention, the structure of lincomycin has been clarified! it deals

009020/191 «

the compound is methyl-6- (trans-4-? propyl-1-methyl-L-prOlylaraino ) -6, e-dideoxy-l-thio-D-erythro-a-D-galacto-octopyranoside following formula:

NS,

It has now been found that lincomycin, isomers and analogs of this compound can be synthesized in which one has a 6-amino-6 ^ 8-dideoxy-I} -erythro- or L-threo-D ^ galacto-octopyranose compound the formula

III

in the Rp and R, hydrogen or Aliyireste with no more represent as 12 carbon atoms, and Y is hydrogen,

009820/1916

^{& AD}

_ ■ 3 -

S-Al kyl with no more than 12 carbon atoms or the grouping -SGB ^ CH ₂ OR ₄ , in which IL is hydrogen or an alkyl radical mxt no more than 12 carbon

Means substance atoms, aylated with a 4-substituted L-proline of the formula _;

II

in which Z is a schatjEender i ^ dröceirbyloxycarbQn ^ grJlippe, which "can be removed by hydrogenolysis, or a trietyl- (triphenylmetiiyi-.), jiiphenylr- (p-me / thoxyphenyl) -methyl-, bis- (p- methoxyphenyl) -pheriylmethyl, benzyl or p-nitrobenzyl radical and R is an alkylidene, aralkylidene or cycloalkylidene radical of the type described below, with the formation of compounds of the formula

OR,

BATH ORIGINAL

in which Z, Y, R, R ₂ and R- have the above meanings; the radical Z is removed by hydrogenolysis and, if appropriate, the resulting compound (formula V, R-, = hydrogen) is alkylated to form a compound of the formula

in which R _{1 represents} an alkyl radical having not more than 20 carbon atoms, a "cycloalkyl radical having 3 to not more than 8 carbon atoms" or an aralkyl radical having not more than 12 carbon atoms, and Y, R, Rp and R have the above meanings.

Examples of alkyl residues with no more than 20 Carbon atoms are methyl, ethyl, propyl, Butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, Undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, Hexadecyl, heptadecyl, octadecyl, nonadecyl and eicosyl radical and their isomeric farms. Examples of suitable

009820/1916

Oycloalkyl radicals are the Gyclopropyl-, Cyelobutyl-, Gyclopentyl-GycloiBxyl-, Cycloheptyl-, Gyclocetyl-, 2-iethylcyclopentyl-, 2, J-dimethylcyclobutyl, 4-methylcyclobutyl and 3-cyclopentylpropyl radical. Examples of aralcyl residues are the benzyl, Phenathyi-, a-Phenylpropyl- and a-Kaphiliyimetnylrest. Examples for Hydrocarbyloxyoarbonylgruppen are tert, -ButQxyoarbonyl, Benzyloxycarbonyl groups of

in which X is hydrogen, a nitro, methoxy group, chlorine or bromine, for example the carbobenzoxy radical (benzyloxycarbonyl), p-l-nitrocarbobenzoxy radical (p-nitrobenzyloxycarbohyl), p-bromine and p-chlorocarbobenzoxy radical (p-bromo- and p-carbonylbenzoxy) radical , as well as phenyloxycarbonyl groups of the formula ^: \ ■ *

in which X _{1 represents} hydrogen, an allyl group or an alkyl group having not more than 4 carbon atoms, such as, for example, the phenyloxycarbonyl radical, p-tolyloxycarbonyl radical, p-ethylphenyloxycarbonyl radical, allyloxycarbonyl radical and the like.

The Att-sgan-gsver-connections of the formula II weH (; a -lurch the following reaction sequence is established:

VII

The resulting product is a mixture of ice and

trans isomer

Ha

Man

and

Trans

G sharp

009820AIIIf

BATH

Γ Ί6206Θ7

The above reactions are carried out by adding a 1 ^ Hydroearbyloxycart) onyl - 4 -] ieto - II-proline (VI) or another: protected 4-keto-L-proline with a, Wittig ¹ see reagent, usually an alkylidene triphenylphosphorane (cf. »Wittig, employees, Ber ,, 87, 1348 (1954); Trippett, Quarterly Reviews, XVII, Fo. 4, p. 4G6 (1963)) and the 4-alliylidene-1-hydrocarbyloxycarbonyl obtained in this way ⁱ -L - proline (VII) hydrogenated in the presence of a platinum catalyst, with the formation of the corresponding 4-alkyl-1-hydrocarbyloxyearbonylpröliiis (II).

■ When carrying out this procedure, the l-hydrocarbyloxycarbonyl-4-keto-L-proline (VI) is added to a freshly prepared Wittig reagent. The Wittig ¹ see reagents used here can be represented by the following formula

be reproduced:,. " _:

VIII

in the row an allcjriide residue with no more than 12 carbon stoffatiJ-.en, a cycloalkylidene radical with, 3 to not iaehr as 8 cabbage nfitoffatoc; en or an Aralkylldenreet with no - Represents more than 12 carbon atoms. Suitable alkylidene, - or aralkyl groups are e.g. the groups:

O O 98 2 07131 6 BATHROOM ORIGINAL

! »! Ethylene, ethylidene, propylidene, butylidene, pentylidene, hexylidene, heptylidene, octylidene, monylidene, decylidene, lindecylidene and üödecylidene and their ioomers, cyclopropylidene, cyclobutylidene, cycloijentylidene, cyclohexylidene, cycloheptylidene, cyclohexylidene, cyclopropylidene, cyclohexylidene, cyclohexylidene Benzylidene, 2-then: / ethylidene, 2-phenylpropylidene and u-naphthylmethylene. The Wittig reagents are obtained by adding an alkyl, cycloalkyl or aralkyltriphenylphofiphonium halide with a base of sodium amide or with sodium or Kaliuinhydrid or the sodium or üaliummetalat of dim ethyl suli'oxyd or the like implements. BeispielBweise the elimination of hydrogen halide resulting from an alkyltriphenylphosphonium to Alkyliaentriphenylphonphoran. £ the preparation of phosphoranes is detail by Trippett, Quart. Rev.XVII, Ho.4, 3.406 (1963) / The reaction is usually carried out in an organic solvent such as benzene, T. oluene, ether, dimethyl sulfoxide, tetrahydrofuran or the like at temperatures between 10 ° C. and the reflux temperature of the reaction mixture. The product thus obtained, a 4-alkylidene, 4-cycloalK: ylidene or 4-aralkylidene

009 8 2 0/19 te

1-hydrocarbyloxyearbonyl-L-proline (VII) is isolated from the reaction mixture in a conventional manner, conveniently by extraction from an aqueous solution. The raw product can be purified in a conventional manner, for example by recrystallization, odorization or the formation of derivatives and recrystallization of the latter; this are, for example ^ a.minsalze as dicyclohexylamine or dergleichenÄ- ^/

The 4-alkylidene- ^ ^ -Cyeloalkylidene- or ^ aralkylidene-1-hydrocarbyloxyaarbonyl-L-proline (VII) is then in the presence of platinum, // which is usually on a support such as carbon or a gj £ nem anion exchange resin such as Dowex-1, a cross-linked folystyrene trimethylbenzylammonium resin, is hydrogenated. A mixture of the cis and trÄs epimers of the 1 -hydrocarbylpxyjaarbonyl-4-subs ± itui ^ rten Ιϊ-proline (Ha and lib) is obtained.

The procedure Jmnn also carried out in the manner be that missing compound III by means of the compound VII acylij ^ i, forming a compound of lOraiel

0 0 9820/1116

BAD OB1G »NAL

R ₂ O

MI HO.

IVa

OH

OR,

and then the alkylidene group with a platinum or palladium catalyst hydrogenated. When using a platinum catalyst, the group Z is retained, while when using a Palladium catalyst, e.g. palladium-carbon, at the same time ·; with saturation of the alkylidene group, hydrogenolysis of group Z occurs.

The starting compounds of the formula III can be obtained in various ways. methyl-b-aruino-6,8-dideoxy ~ l-thio-D-erythro-u-D-galacto-octopyranoside, also known as Methyl-tt-thiolincosaminide or i> iIL is called obtained by subjecting lincomycin to hydrazinolysis; Ethyl-o-amino-ö.B-dideoxy-l-thio-D-erythro-u-D-galacto-octypyranosJLd, also called Ethyi-u-thiolincosaminide : or KHL is obtained by hydrazinolysis j of mncomycin 0 »an antibiotic that is produced

009820/1316

ORIGINAL

■ if the lincOinycin fermentation .gomä'i- B.S. ^ i'ätent 3,086 * 912 in the presence of ationine takes place /.: ■ "_; - ,. \.; -"

The hydrazinolysis is carried out 2iweofettäßi |; eTv? E ± oe ,. by cooking the antibiotic at Miekfluk Jc for about 20 hours or more with an over-the-counter. The ^ .nv -'- 'non-6-Ämino-D _i 8-dideoxy-1-tbia-D-srytlirO'-uD-ibalacto-octopyranoside can be isolated by distilling off excess hydrazine and adding the trqduict like i? inoB. polar solvent like ethanol crystallizes * ν .. 'V ■', - -------

'tl / 3OtO-Oetopyraftoside can bend to the following

hL-rge «" 41t be: -.;

SAD ORJGiWÄL 00982071916

Reaction sequence A-I

'

Ilia

CH ₃

MTL

HO-AcNH- IX

- ο,

HO λ

N

SCH ₃

OH

O-C-

CH ₃

IHb

alkyl

(J) CSO-

CH ₃

AcNH-

o-cs |)

CSC-AcNH-

(J) CSO

■ ι XIII

ο-οοψ

CH ₃

AcI

J) CSO XIV

/ ι I

NL / Salkyl

OCOJ)

0-0 ^ 8-2 0. / 1916

In the above formulas, Ae represents an alkanoyl or aralkanoyl group having not more than 12 carbon atoms and i material. Examples of these are the formyl, acetyl, propionyl, ßutyryl, valeryl, hexanoyl, undecanoyl, dodecanoyl radical and isolated forms thereof, the benzoyl, phenylacetyl, 3-phenylpropionyl, 4-phenylbutyryl, 5-phenylvaleryl radical and isomeric forms thereof.

The present procedure is carried out as follows; liotüyl-u-thiolincosaaiinid v / i-rd IT-aoylated, das Methyl-N-acyl-iu-thiolinoosaiüinid (IX) is then with Thiobenzoyl chloride-a-cylated, the resulting thiobenzoylaster (X) is brominated to give the compound XI, which is converted to the compound XII by treatment with a base is convicted. Treatment of compound XII with weak alkali (aqueous carbonate bicarbonate solution) leads to compound XIII, which on alkylation with oineDi alkyl halide gives compound XIV. the Hydrolysis of the benzoyl and thiobenzoyl groups as well Eydrazinolysis of the N-acyl group leads to the compound IHb * In the same way, connections in which Y a 2-hydroxyethyl or 2-alkoxyethyl radical is produced are made by reacting the compound XIII with ethylene chlorohydrin or a 2-alkoxyethyl halide.

009820/1916

Reaction sequence A-2

CHj

Ac-NH-HO

CH.

O-

R ₀ SH

HO-Ac-NH-

NS.

OH

CH ₃

HO

AcNH HO

OH

HO / H) H _{SR (}

OH

OH

SRo

OH

In the above formulas, ac can be an alkanoyl or aralkanoyl radical of the type described above or an acyl radical of the following formula:

RH '

or

- 14 -

009820/1916

BATH ORIGINAL

in which R, R, and Z have the above meaning R Can be an alkyl radical with 2 to not / t more than 12 carbon atoms or a radical -OH ₂ GH ₂ OR *, in v? el cn ear R. has the above meaning. The agaric glycoside A, w-jlohes is advantageously lincoriyein, ■ is subjected to a capping with an aercaptan of the formula -R ₀ SIi with formation of the dithioacetal B, which in turn is due to the action of heat and / or acid ^ u is cyclized in thioglycoside Ö «."-.;.

009 820/1 9 16 BAD ORIGINAL

Reaction sequence A-5:

CH,

CH,

HO -

AcEH HO

OH

OH

Br,

/ fiJGH, '

HO i AcNH

OH

Ji'a

CHO

In the above formula, Ac and R _{0 have} the meaning given for the reaction sequence A-2. The Ausgar% s connection A confused! in the form of a soluble salt, for example the hydrochloride, dissolved in water, whereupon bromine is added with cooling to between approx. -10 ° and 20 ° C. bromine. A suitable procedure is as / ϊ one ax aqueous Lor;? U ig cools down to about 0 ⁰ C and the Bro ^ s is added dropwise "The stectjioiLetrische

Brooimenge is a & ol per KoI starting compound, but larger or smaller amounts can be used. It is advisable to use a small excess of bromine of b-20 % .

00982071916

BATH ORIGINAL

The thioglycoside is converted into a sugar F in which the pyranos'eforin Fa is in equilibrium with the aldose form Fb. This sugar can optionally be isolated, but this is usually neither necessary nor desirable. In the presence of acid, e.g. hydrochloric acid or another strong non-oxidizing acid such as p-toluenesulfonic acid or sulfonic acid anion exchange resins, the mercaptan R ₀ SH reacts with the sugar F to form the thioglycoside G. An effective conversion occurs when the mercaptan is used R ₀ SH is introduced into the aqueous solution obtained in the bromination, and the resulting system, which is usually two-phase, is treated (because of the insolubility of the mercaptan) with gaseous hydrogen chloride or concentrated hydrochloric acid, advantageously with cooling to a temperature between approx -10 ° and 20 G. A suitable procedure consists in cooling the reaction mixture to 0 G and introducing hydrogen chloride gas. It can be advantageous, especially in the case of longer-chain mercaptans, to partially dissolve them by adding tetrahydrofuran to the reaction mixture,

009820M916

BATH

For this purpose, 15 to 20 parts of tetrahydrofuran are used per part of water. The temperature of the reaction mixture can rise during the addition of hydrogen chloride, but expediently not to more than about ρ5 G. After cooling to about 25 ° C., the tetrahydrofuran is removed in vacuo. The reaction mixture can be worked up in the usual way, for example by solvent extraction at acidic pH, solvent extraction at basic pH, fractional liquid-liquid extraction such as countercurrent distribution or partition chromatography, Λ-crystallization or the like. At the same time, small amounts of dithioacetal of the formula b (reaction sequence A-2) can be formed which, after separation, can be cyclized as described above, with the formation of the desired thioglycoside

The ß-epiiueren of the above compounds can on can be produced in the following way:

009820/1916 bad original

Reaction sequence B

ΪΧ

methyl-N-acyl-athiolincosamine

Ad-NH

XV0

Ac * -NH XVIß

Ac'-NH. XVIII

OH • alkyl-ß-thloline cosamine ld Alkyl-N-acyl-ß-thloline cosatinide

2 0/1916

BATH

Ac and Ac- represent alkanoyl or aralkanoyl radicals with represent no more than 12 carbon atoms.

The procedure is carried out as follows: han treated l'lethyl-W-acyl-u-thiolincossuuinid (IX) rait mercury chloride in warm aqueous solution, over aan a mixture of ö-acylamino-ojH-di-ieoxy-D-erythrou-D-galacto-octopyranorje or N-acyl-u-lincosamine (XVa) and ii-acyl-β-liricosamine (XVβ) is obtained; then acylated this tread sch with an acyl hai οjenid or acid anihydride forming a mixture of II-acyl-1,2,3, -4,7-penta-O-ooylu- and β-lincosaiuin (XVa and XVIß); then b-.hando.lt one the compound IVIa or XVIß or a mixture of these compounds with hydrogen bromide in acetic acid, whereby 6-acylamino-2,5,4 "r7-te-tra-0-acyl-lu-brou: -l,.,.:; - trideoxy-D-erythro-D-galaoto-octopyranose or W-acyl-2,5 »4,7-tetra-ü-acyl-lu-orom-1-aeoxylincosanine (XVlI) is obtained; the connection XVIl is then Kaoheiriander with thiourea, a G-e.Bioch made of potassium carbonate and sodium bisulfite treated in water and an alkyl iodide, whereby alkyl-li-acyl-2,3,4,7-tetra-0-acyl-ß-tniolincoca..iinid (XVII) is formed;

009820/1916

The compound XVIII is then treated with dry ammonia gas in methanol to form an alkyl-N-acyl-thiolincosaminide (XIX). The hydrazinolysis of the compound XIX or the compound XVIII leads to the alkyl-ß-thiolincosaminide (illc). In the same way, compounds in which Y (Poruel III) is a 2-hydroxyethylthio or 2-Alkoxyäthylthipgruppe is, are prepared by Implementation of the compound XVII with ethylchlorohydrin or a 2-alkoxyethyl halide.

Compounds of the formula III, in which Y represents hydrogen, can go through the following reaction stages getting produced:

0 0 9 8 2 0/1916 "BAD original

Reaction sequence C

CH ₃

HO · AcNH.

IX

T & ■

NK SCH ₃

OH

'CH ₃

AcNH

XXII CH ₃

HO AcNH-

CH ₃

CH ₃

XXI

CH ₃

XXIII CH ₃

XXIV

OH

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The above procedure is carried out as follows: methyl-H-acyl-a-thiolincosarainide. (IX) * (or rtethyl-W-acyl-ß-thiolincosaminide) is treated with acetone in the presence of sulfuric acid, whereby l'iethyl-N-acyl-3,4-0-if5: jrop, yliden- «.- thiolincosaminide (XX) is formed, which can be desulfurized with Raney-i-nickel in ethanol, with the formation of o-acylamino-'jj ^ -O-isopropylidene-1, bjb-trideoxy-D-erythro-D-galacto-octopyranose or IJ- Acyl-3,4-δ-isopropylidene-1-deoxylincosamine (XXI). The desulfurization is expediently carried out by boiling on RUcicflu; in methanol packed with perforated Raney nickel for 2 to 24 hours. The catalyst is then filtered off and the filtrate is evaporated, leaving a residue which can be purified in a conventional manner, for example by recrystallization from organic solvents. The iso / rop ^ lidenöi'i'i'i'i'i'i'i can then be removed by mild acidic hydrolysis, for example with 80 % aqueous acetic acid or -a dilute kineralsäure such as hydrochloric acid or Scr-vefGlftire, where.: An li Acj'l-l-deoxylincosarnine (XXII) obtained; ax K acyl group can be removed by hydrazinolysis

^009820/1916

where 1-deoxylineosamine (XIII) is formed. Aie hydrazinolysis can also be carried out first, whereby 3,4-0-Isopropylidene-1-deoxylincofiamine (XXIV) is formed. Advantageously, however, the isopropylidene group is removed first.

The Ausgarigsverbindungen of formula III, in which R ₂ and / or R, is an alkyl radical with not more than 12 carbon atomsj. Can be obtained in the following way:

009820/1916

^ß AD ORIGINAL

Reaction sequence D

XXVI

XXVII

Rs-C-NH- _ * »

Rs-C-

OH

CH ₃ -

XXVIIIb 0? ^H3 XXVIIIc

j) R _e C Rs-C-ίΙΗ-

HO-H ₃ N. XXIXb

Location

- 25 -

009820/1916 BAD

In the above scheme, RcCO - can represent an alkanoyl or aralkanoyl radical with no more than 12 carbon atoms; Rg and Ry can be different alkyl radicals if iodine compounds XXVIIIa and / or XXVIIIb have been treated with alkylating agents which differ from the putties used for the alkylation of the compounds XXVII. Rg and R ₇ in the compound XXVIIIc can thus be different alkyl radicals.

The above process is carried out as follows: An δ-acylamino-ojÖ-dideoxy-D-erythro-D-galacto-octopyranose compound of formula XXV is treated with dry acetone in the presence of an acidic catalyst, for example sulfuric acid, which is a mixture of the compound XXVII and its oxazoline derivative XXVl obtained; Hydrolysis of the compound XXVI with hot water gives the compound XXVII, which is reacted with an alkyl halide, for example an alkyl chloride, alkyl bromide or alkyl iodide having not more than 12 carbon atoms, in the presence of a base, whereupon the three products thus obtained (X / IVIIIa, XXVIIIb and XXVIIIc) separates; by hydrolysis of the compounds XXVIII under mild conditions, for example with 80 % aqueous acetic acid or dilute mineral acid such as salt

BATH ORIGJWAL 009820/1916 ^L

acid or acid, the isopropylidene residue is removed, and by dehydration with hydrazine, the N-aoyl group is removed and the compounds ΧλΙΧ are obtained. If Y in the compounds treated in this way is a 2-Hydroxyäthyltliiogruppe or uses aan 2-Hydroxyäthyl-6-amino-6, b-dideoxy-7-O-.nethyll-tnio-l) -erythro-uD-galacto-oct'opyranosid, which is also known as 2-hydroxyethyl-u-tuiocel ~ estosaininide, and can be prepared by hyarazinolysis of celesticetin, an antibiotic obtainable according to US Pat with the formation of a linear alkoxyethylthio group, in which the alkoxy group contains no more than 12 carbon monoxide oxides. If necessary, the 2-hydroxytiiiogrUj.ie ceo can be used with the help of a trityli group. Compounds XXV or XXVII, in which Y is a 2-hydroxyethylthio group, can be mixed with trityl chloride (triphenylethylchloride), chlorophenyl- (p-methoxyphenyl) -. -phenyl.nethan implemented vari ^ n, untt-i Lr-vandlun ^ from Y-ii. a 2-trityloxyethylthio group, uiii di «'' tri ^ yl group, then becomes after the alkylation

0098 20/1916 BAD O «G» NAL

removed by mild hydrolysis, for example with 80 $ aqueous acetic acid. The 2-hydroxy group can be alkylated with a different alkylating agent. When carrying out the above process, the 6-acylamino-öje-dideoxy-D-erythro-D-galacto-octopyranose compound of Forxael XXV is suspended in dry acetone. An acidic catalyst such as sulfuric acid, p-toluenesulfonic acid, o-toluenesulfonic acid, p-ethylbenzenesulfonic acid or the like is added to the suspension, sulfuric acid being the preferred catalyst. The acetone is usually used in large excess, for example in 20 to 200 times the volume, based on ax compound XXV. The acid catalyst is used in Kengen of 0.25 to 5 wt .- ^ ₅ to the .aceton used. The reaction can take place between 10 ° C. and the reflux temperature of the solution, but is usually carried out at room temperature. The reaction time is between 15 minutes and 6 hours, depending on the reaction used?; - temperature, after which the reaction mixture is neutralized to complete the reaction. The organic salts which precipitate during the neutralization are filtered off and the mush is evaporated, whereby a crystalline solid is formed

009820/1916 BATH ₀ R / g / _NA l

containing mixture obtained. The .mix is with the help von Waseer into a water-soluble and a water-insoluble Fraction dismantled; the water-soluble material is the compound XXVII, while the insoluble Material consists of the oxazoline derivative thereof (XXVI).

The water-insoluble oxazoline derivative (XXVI) can be converted into the compound XXVII by heating in water. After the hydrolysis has ended, the product XXVII is obtained by evaporating the solution until crystallization begins.

Compound XXVII is alkylated with an alkyl halide, preferably an alkyl chloride or bromide or iodide, in the presence of a strong base · Als Alkylating agents are methyl iodide, methyl bromide, Ethyl iodide, ethyl bromide and propyl, butyl, isobutyl, Pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyi, Dodecyl chloride, bromide and iodide as well as branched chain ones Al-vinyl chlorides, bromides or iodides with up to 12 carbon atoms. Alkoxides such as sodium, iietnylate, Potassium methylate, potassium isopropoxide, potassium tert-butoxide, Use sodium tert-butoxide and sodium amide or the like. The alkylation can be at a Temperature between 0 and 50 0 using inert

009820/1916 BAD

Solvents such as benzene, toluene, dioxane, tetrahydrofuran, or the like. When the reaction is complete (usually between 1/2 and 3 hours), the reaction gushes filtered to remove solids such as sodium or potassium chloride, to remove bromide or iodide, and the piltrate will be ir :; Concentrated in vacuo to a syrupy residue. The three products in it, namely the connections XXVIIIa, XXVIIIb and XXVIIIc are separated in a conventional manner, for example by chromatography or by countercurrent distribution. The removal of the N-acyl and isopropylene groups are carried out as in the Reaction sequence C described. Of course, 2-0-, 7-0- and 2,7-di-O-alkylates can be used according to the Formulas XXIXa, XXIXb and XXIXc are used in the reaction sequences A, B and C, using the 2-0-, 7-0- and 2,7-di-O-alkylates of the corresponding compounds are obtained.

The 7-0-epimers of the compounds III, i.e. the o-alaino-öjS-dideoxy-L-threo-D-galacto-octopyranose compounds, can be obtained by using epi-MTL (j> iethyl-6-amino-o, 6-dideoxy-lthio-L-threo-tt-D-galacto-octopyranoside) as the starting material the

009820/1916

IHd

begins. Epi-MTL can go through the following reaction stages

will be obtained:

CH ₃

CH ₃

IXa (I)

CH ₃

SOI®

H ₃

SCH «CH ₃ .

HC

PH I.

■ · - 31 - -. . ; 009020/1916 "- BAD OR! G« NAU

In the above formulas, Ac can be an alkanoyl or aralkanoyl radical having not more than 12 carbon atoms or a represent trans-4-propyl-1-methyl-L-prolyl radical in which Case IX lincomycin (I) and XXXII epilincoruycin / S- (trans-4-propyl-l-methyl-L-prolylajnino-6,8-dideoxy-l-thio-L-threo-u-D-galacto-octopyranoside ^ represents.

The above reaction sequence can be applied to any 6-amino-6,8-dideoxy-D-eerythro-D-galacto-octopyranose compounds of Forinel III (or N-acylates thereof) are applied.

When carrying out the above process, an N-acyl derivative of a 6-atnino-6,8-dideoxy-D-erythro-D-galacto-octopyranose compound of the formula III, preferably lincomycin, is used as the starting material; for example, lincomycin (I) with acetone around the formation of the 3,4-0-isopropyliaen-1) erivats XXa, in the manner described for the reaction sequences C and D, with lincomycin as starting material, the formation of the oxazoline XXVI is kept to a minimum. The 3'4-0-isopropylidene-linoomycin XXa is then oxidized with chromic acid to form the 7-dehydro-i »erivats XXX, which in turn is reduced with borohydride to form a mixture of 3 % 4-Ü-isupprop, ylidene-lincomycin and 3 »4-0-isopropylidene-epilincor: jycin (XXXI). The two

009820/1916 SAD

Epimers can be separated by liquid-liquid extraction, for example by countercurrent distribution or Partition chromatography, or by adsorption or stepwise chromatography, or the mixture can be converted into a Mixture of lincomycin (I) and epilincomycin (XXXII) can be transferred, and the two epimers are more similar Way separated. Removal of the 3,4-0-isopropylidene group takes place by mild hydrolysis, as for the reaction sequence G, whereby epilincomycin. (XXXII) is obtained, which in turn is deacylated with hydrazine to form the desired methyl-epi-u-thiolincosaminide (methyl-6-v amino-e ^ -dideoxy-l-tliio-L-threo-iA-D-galacto-octopyranoside), or hydrazinolysis is carried out first and the isopropylidene group is then removed.

The protective group Z in the compounds of Formula IV is removed by hydrogenolysis using a palladium catalyst. The palladium. is usually located on a carrier, for example auJ coal, used. Hydrogenolysis can be carried out in any done conventionally.

The proline nitrogen in the compounds of the formula V, in which R _{1 is} hydrogen, can be mixed with an alkyl halide,

^009820/1916 BADOB, G.NAL

for example an alkyl iodide alkylated «earth. Mild conditions are used here than for the alkylation of the 2-0 and 7-0 positions. N-alkylation they can not without strong base and at room temperature or at most after heating to about 50 ⁰ C, take place. It is expedient to alkylate with reacting the compound with an oxo compound (an aldehyde or ketone) and hydrogenation of the resulting adduct. The hydrogenation can be carried out with palladium or platinum or any hydrogenation catalyst that saturates olefinic double bonds Cycloalkylidene group with 3 to not more than 8 carbon atoms, or an aralkylidene group with not more than Example represented by 12 carbon atoms for such aldehydes are formaldehyde, acetaldehyde, propionaldehyde,

Butyraldehyde, acetone, isobutyl methyl ketone, benzaldehyde,

phenylacetaldehyde, hydrocinnamaldehyde, acetophenone, propiphenone, butyrophenone, 3-methyl-4-phenyl-2-butanone _i 2-methyl- 5-phenyl-3-pentanone, 3-cyclopentane propianaldehyde, cyclohexaneetaldehyde, cycloheptane carboxaldehyde, 2,2-dimethylcyelopropyl

BAD ORIG'NAL 009820/1916

acetaldehyde, 2,2-Dimethylcyelopröpylmethylketon, Cyclopentylmethylketon, Cydobutylaethylketon, Cyclobutanes, Cyclohexanone, 4-methylcyclohexanone and the like.

The de and trans epimers can be separated by partition or stepwise chromatography * The separation is best done in the procedural stage represented by the formulas IY or V. They can also form epimers by starting from the trans or cis form of the 4-substituted L-proline (formula Ha and lib).

Acid addition alβ the free bases of the compounds V, in which B ^ is hydrogen, alkyl, cycloalkyl or aralkyl *, can be obtained by neutralizing the base with a corresponding acid at pH values below about 7.0, preferably at pH about 2 to 6 Acids suitable for this purpose are hydrochloric acid, sulfuric acid, phosphoric acid, thiocyanic acid, fluosilicic acid, hexafluoroarsenic acid, hexafluorophosphoric acid, acetic acid, succinic acid, citric acid, lactic acid, maleic acid, fumaric acid, pamoic acid, glycolic acid, palmitic acid, glucic acid, glyceric acid _, tartaric acid , Lauric acid, stearic acid, salicylic acid, 3-phenylsalicylic acid, 5-phenylsalicylic acid,

009820/1916

BATH

3-methylglutaric acid, benzoic acid Orthosulf ©, Cyclohexansulfamsäure, Cyelopentanpropionsäure, 1.2 ~ cyclohexanedicarboxylic acid, ^ -Cyelohexenearbonsäure, octadecenylsuccinic acid, octenylsuccinic acid, Methansulfoneäure, Benaolsulfonsäure, Helianthsäure, Reinecke acid ^B-1, dimethyldithiocarbamic acid, cyclohexyl sulfamic acid, Hexadecylsulfamsäure, Octadecylsulfamsäure, sorbic acid, Honochloreseigeäure , Undecylenic acid, 4 * - hydroxyazobenzene-4-sulfonic acid, octyldecylsulfuric acid, picric acid, benzoic acid, cinnamic acid and the like.

The acid addition salts can serve the same purposes used like the free bases or they can serve to purify the bases. For example, the Convert the free base into an insoluble salt, such as the picrate, which is then purified, for example by Solvent extraction and washing, chromatography, fractionated liquid-liquid extraction and crystallization, whereupon the free base is regenerated by treatment with alkali or with another salt with metathesis. Also can the free base can be converted into a water-soluble salt such as the hydrochloride or sulfate, and the aqueous Solution of the salt cannot be different with water

ORIGINAL BATHROOM 009820/1916

miscible solvents are extracted before the Regeneration of the free base takes place.

The compounds of the formula V in which is hydrogen, alkyl, cycloalkyl or aralkyl can serve as buffers or acid antagonists. The compounds of the formulas 17 and V react with isocyanates to form urethanes and can be used to modify polyurethane resins. Me long-chain compounds, i.e. those in which R _{1 is} an alkyl radical with more than carbon atoms, have surface-active properties and can be used as wetting and emulsifying agents. On condensation with formaldehyde, the thiocyanic acid addition salts produce resinous materials which, according to US Patents 2,425,320 and 2,606,155, are suitable as pickling agents. The free bases are also good carriers for toxic acids.For example, the addition salts with fluorosilicic acid are suitable as moth repellants according to TT.S. patents 1.915 * 334 and 2.075.359 and the hexafluoroarsenoic acid and hexafluorophosphoric acid addition salts are agents for combating parasites according to U.S. Patents 3122 * 536 and 3,122,552.

009820/1916 BAD ORIG'NAL

They are close to analogues of lincomycin, i.e. compounds in which HH- is a trane-alkyl radical with no more as 6 carbon atoms, R ^ a methyl or ithyl group and Y represents an α-alkylthio group having not more than 6 carbon atoms, have bactericidal properties that are comparable to those of Lincomyoin; the compounds can be used for the same purposes as lincomycin will »The other inalogues and isomers have similar ones bactericidal properties, but to a lesser extent; they can be used for the same purposes as lincomycin, in those cases where the application larger amounts is not disadvantageous.

In the following examples, parts and percentages relate to weight; the solvent ratios refer to volume, unless otherwise stated.

BAD ORIGINAL 009820/1916

Example 1:

Methyl-6- (cie- and trans-4-alkyl-l-alkyl-L-prolyl-amj.no ) -6,8-dideoxy-1-thio-D -erythroa-D-galacto-oetopyranoside (Methyl-N- (cis- and trans-4- & lkyl-1-alkyl-L-prolyl) -athiolincosaminide).

R + B ₁ «alkyl

XXXIV

A-I. 4-butylidene-l-carliOtoeneoxy-L-proline and its Dioyclohexylamine salt K.

19 s of sodium hydride in 53% suspension in mineral oil were heated with 350 ml of diethyl sulfoxide at 70-75 ° C. until the reaction had ended (about 30 minutes). After cooling to 32 ⁰ C wurden.16,2 g butyltriphenylphosphonium bromide was added and the resulting mixture was stirred for 1 hour to complete ümeetsung Aicher

009820/1916

BATH

to deliver. Then a solution of 26 g of 4-keto-1-carbobenzoxy-L-proline in 100 ml of dimethyl sulfoxide was added and the resulting mixture was heated to 70 ° C. for 3 hours. The reaction mixture was then ^{cooled to 25 °} C. and 1 liter of 2.5% aqueous potassium bicarbonate solution was added. The mixture was washed twice with 700 ml ie ether and the ether was discarded after jerk extraction with 150 ml 2.5 # strength aqueous Ealiumbicarbonat. The beer carbonate solutions were combined and acidified with 4η hydrochloric acid. The acidified mixture was extracted four times with 500 ml of ether each time. The combined extracts were washed successively with 250 ml of water, three times with 250 ml of saturated aqueous sodium bisulfite solution and with 250 ml of water and dried over anhydrous sodium sulfate. When the solvent was evaporated off in vacuo, 24 g of an oily residue were obtained, which consisted of 4-butylidene-1-carbobenzoxy-L-proline.

This residue was dissolved in 31 ml of acetonitrile and treated with 18 ml of dicyclohexylamine and refrigerated. The crystals that formed were collected, washed with aceto-

0098 20/1916

washed nitrile and dried in vacuo, 21 g (46.8 fi) of the crystalline dicyclohexylamine salt having a melting point of 136-14O ⁰ C being obtained. After two recrystallizations from acetonitrile, an analytically pure sample with a melting point of 142-144 ° C. was obtained; optical rotation Z " ^tt -7 _D >> -4 ° (c = 0.99 in CHCl ₃ ).

Analysis: Calculated for ° 29 ^Η 44 ^ 2 ^ 4 ^:

Ct 71.86; H: 9.15; H: 5.78;

found:
C: 71.69; H: 9.30; N: 5.74.

10 g of the dicyclohexylamine salt were mixed with ether and excess 5 # aqueous potassium hydroxide solution shaken until no solid remained. The layers were separated and each was backwashed.

The aqueous-alkaline layer was combined with the washing water of the ether layer and acidified with 4 η-hydrochloric acid, The mixture was extracted several times with ether and the ether extracts were combined and dried over sodium sulfate and evaporated in vacuo to give 6.3 g (95 #) of 4-butylidene-1-carbobenzoxy-L-proline received as oil.

. . BATH ORIGINAL 009820/19 16

A-2. 4-pentylidene-l-carbobenzoxy-L-proline and its Bi cyelohexylamine salζ.

The procedure of Part AI is used, but replacing the butyltriphenylphosphonium bromide with pentyltriphenylphosphonium bromide; this gives 4-pentylidene-1-earbobenzoxy-L-proline and its dicyclohexylamine salt. The free acid was in the form of an oil, whereas the dicyclohexylamine salt, after recrystallization from acetonitrile, had a boiling point of 124-128 ° C .; optical rotation fu-JQ '· -6 ° (c «0.762 in CHCl ₅ ).

Analysis: Calculated for C3 _O ^H 46 ^N 2 ° 4 ^{S Ci72} » ² 5; H: 9.30; Hi 5.62;

found: 0: 72.38; H: 9.52; H: 5.97.

A-3. 4-hexylidene-1-earbobenzoxy-L-proline and its Dicyclohexylamine salt.

The procedure of Part A-I is used, but replacing the Butyltrlphenylphosphoniumbromids by hexyltriphenylphosphonium bromide; this gives 4-hexylidene-1-carbobenzoxy-II-proline and its dicyclohexylamine salt. The free acid consists of an oil, the dicyclohexylamine salt of a solid, which after recrystallization from acetonitrile showed a melting point of 109-111 C; optical rotation / "aJ7j> s -7 (c == 0.941 in CHCl ^).

■ ". ? ♦ ■ =. .-. ^8AD

009.820 / ^ 16

Analysis t Calculated for 031H ₄₈ N ₂ O _4: C: 72.62; H: '9.44? H: 5.46;

found: C: 72.70; H: 9.43; H: 5.71.

A-4. 4-octylidene-l-earbobenzoxy-L-proline and its Dicyelohexyl aminesals.

One works according to the procedure of Part A-I, but with replacement of the butyltriphenylphosphonium bromide by octyltriphenylphosphonium bromide; 4-octylidene-l-earbobenzoxy-L-proline and its bicyclohexyl- amine salt obtained. You free acid is an oil, das Di cyclohexylamines ^, a solid that after Recrystallization from acetonitrile at 113-118 ⁰ C melts; optical rotation f a_7 _D : -11 ° (c = 1.020 in CHCl ₅ ).

Analysis «Calculated for C ₅₅ H ₅₂ H ₂ O ₄ SC: 73.29; H: 9.69; H? 5.18;

Found: C: 73.32; H: 10.06; N: 5.28.

BI. 4-butyl-1-earbobenzoxy-L-proline.

The oil obtained according to A-I was dissolved in 200 ml of methanol hydrogenated over 2.1 g of 10 SC platinum / Dowex-1 catalyst at 2.8 atmospheres of hydrogen pressure. The catalyst was filtered off and the piltrate was evaporated, whereby one 6.3 g of 4-butyl-1-carbobenzoxy-L-proline in the form of an oil received. The product contained approx. 2 parts of ice-4-butyl-

009β20 / $ 1 $ 1

BATH ORIGINAL

earbobenzoxy-L-proline compound per one part of trans compound.

B-2. 4-P entyl-1-carbobenzoxy-L-proline.

In the hydrogenation of the after. A-2 obtained oils following the procedure in Part B-I, 4-pentyl-1-carbobenzoxy-L-proline was obtained in the form of an oil.

B-3. 4-hexyl-1-carbobenzoxy-L-proline.

Hydrogenation of the oil obtained in A-3 by the procedure of Part B-I gave 4-hexyl-1-earbobenzoxy-L-proline in the form of an oil.

B-4. 4-octyl-1-carbobenzoxy-L-proline.

Hydrogenation of the oil obtained in 1-4 by the procedure of Part B-I gave 4-octyl-1-carbobenzoxy-L-proline in the form of an oil.

C. Methyl-e-amino-oiS-dideoxy-1-thio-D-erythro-tt-D-galacto-octopyranoside (Methyl-a-thiolincosaminide).

A solution of 40 g of lincomycin (U.S. Patent 3,086,912) in 20 ml of hydrazine hydrate (£ 98-100) was made Refluxed for 21 hours; Excess hydrassin hydrate was then added in vacuo under nitrogen Removed steam bath temperature leaving a residue. The residue, a pasty crystal mass, was cooled,

009820/1916

ÖAD ORIGINAL

added acetonitrile and the mixture was stirred, until the crystals were suspended. They were then filtered off and washed with acetonitrile and ether. After drying the product in vacuo at room temperature, 21 g (84 #) of white, crystalline methyla-thiolincosaminide were obtained. Recrystallization was carried out by dissolving the methyl-α-thiolincosamine in hot dimethylformamide and adding an equal volume of ethylene glycol dimethyl ether.

Methyl-a-thiolincosaminide has a melting point of 225-228 ° C; optical rotation £ α _7 _D * + 276 ° (c = 0.768 in water); pKa ¹ = 7.45.

Analysis: Calculated for CqHj ^ qNO ^ S: C: 42.7; H: 7.56; N: 5.53;

S: 12.66;

found: C: 42.6; H: 7.49; N: 5.75ί

S: 12.38.

D. Methyl N- (4-alkyl-1-carbobenzoxy-L-prolyl) - «- thiolincosaminide.

Z Z

ι »

+ MTL

HR J Γ °° ^0E Y— ¥ COMTL

MR

00982071916

BATH

D-I Hethyl-N- (4-butyl-l-earbobenzoxy-L-prolyl) -α-thiolincosaminide

To a solution of 6.3 g of 4-butyl-l-oarbobenzoxy-L-proline (oil from Part A) in 175 ml of distilled acetonitrile, cooled to 0 °, 3.46 ml of triethylamine and then 3.34 ml of isobutyl chloroformate were added . The mixture was stirred at 0 C (+ 3) for 15 minutes; then a solution of 6.2 g of ethyloxthiolincosaminide (HTL) from Part B in 85 ml of water was added and the reaction mixture was ^{stirred at 0} ° C. for half an hour and at 25 ° C. for one hour. The reaction product was then filtered off and dried to give 4.57 g (37.7 #) of methyl-N- (4-butyl-1-earbobenzoxy-L-prolyl) -α-thiolincosaminide. The mother liquor was concentrated in vacuo and an additional 4.25 g (35.2%) of product was recovered. Recrystallization from acetonitrile gave crystals of ethyl-H- (4-butyl-1-earbobenzoxyl-prolyl) -a-thiolinco8aminide with a melting point of 194-196 C. Further recrystallization from acetonitrile gave an analytically pure sample with a melting point of 197.5-200 C won; optical rotation / ü_7 _D : + 111 (c = 0.98 in HeOH).

0 0 9 8 2 0/1 916 Original bathroom

Analysis: Calculated for C ₀ ZrH _11n H ₀ OQS: C: 57.75; H: 7.46; Hs 5.13;

^{2b 40 Z ö} S: 5.95;

Found: C: 57.58; H: 7.16; N: 5.50; S: 6.07.

D-2. Methyl N- (4-pentyl-1-earbobenzoxy-L-prolyl) -octhioline cosaminide.

If one works according to the procedure of Part D-I, but replacing the 4-butyl-l-carbobenzoxy-L-proline by the 4-pentyl-l-carbobenzoxy-L-proline compound, Part B-2, 80 you get methyl-N- (4-pentyl-l-carbobenzoxy-L-

prolyl) -u-thiolincoeaminide of melting point 191-193 C; optical rotation C ^a i * ⁺ 108 ° (c * 0.722 in MeOH).

Analysis: Calculated for C 7 ^H 42 ₂ 2 ^{0 Ii} 8S: C: 58.46; H: 7.63; Jf: 5.05;

Found: C: 58.32; H: 7.52; H: 4.95.

D-3 · methyl-H- (4-hexyl-1-carbobenxoxy-L-prolyl) -α-thiolincosaminide. '

If one works according to the procedure of part DI, but replacing the 4-butyl-l-carbobenzoxy-L-proline with the 4-hexyl compound (part B-3), one obtains methyl-N- (4-hexyl- l-carbobenzoxy-II-prolyl) -a-thiolincosaminide of melting point 176-180 ° C; optical rotation / "a_7 _D : + 103 ° (c * 0.951 in methanol).

00 9 820/1916

'"■ BAD ORIGINAL

Analysis: Calculated for C ₂₈ H ₄₄ N ₂ O ₈ S:

Oi 59.13; Hi 7.80; N: 4.93; S: 5.64; found; C: 59.16; H: 7.46; »S 5.09; S: 5.96.

D-4. Methyl N- (4-0etyl-1-carbobenzoxy-L-prolyl) -uthiolincosaminide.

If one works according to the procedure of part DI, but replacing the 4-butyl-1-carbobenzoxy-L-proline with the 4-octyl compound (part B-4), one obtains methyl-N- (4-octyl- l-carbobenzoxy-L-prolyl) -a-thiolincosaminid of melting point 181-202 ⁰ C; optical rotation C oil J : + 99 ° (c m 1.083 in methanol).

Analysis: Calculated for Ov ₀ H ₄₈ N ₂ OgS:

0: 60.38? H: 8.11; N: 4.70; S: 5.37; found: C: 60.35; H: 8.08; N: 4.73 *

B. Methyl N- (4-alkyl-L-prolyl) -u-thiolincosaminide hydrochloride.

I, N .HCl

COMTL 'Y— Y COMTL

MR

9820/1916

SAD ORIGINAL

BI. Methyl-Jf- (4-butyl-L-prolyl) -a-t-molineosaminide-1 hydrochloride.

A solution of 7.8 g of methyl N- (4-butyl-1-earbobenzoxy-L-prolyl) -tt-thiolincosaminide from Part B in 200 ml of methanol was poured over 2 g of ten percent palladium-carbon catalyst under 2.8 atmospheres Shaken under hydrogen pressure for 17 hours. The catalyst was filtered off and the solution was evaporated in vacuo. The residue was dissolved in a mixture of 20 ml of acetone and 20 ml of water and acidified with 6N hydrochloric acid. When diluting with 4 volumes of acetone, methyl N- {4-butyl-L-prolyl) -a-thiolincosaminide hydrochloride precipitated, which was filtered off and dried. After drying at 35 ° in vacuo, the crystals weighed 4.7 g and melted at 188-194 °. An analytically pure sample obtained by recrystallization from acetone melted at 197-199 °; £ ~ ^α 7ο * ^{+ 15} °° ( ^water * ° * 0.89). Analysis: Calculated for C ₁₈ Hz ^ NgOgS.HCl:

C: 48.80, · H: 7.96? If: 6.32; Si 7.24; found: (corrected for 5.3 S * water);

C: 48.58; H: 8.19; N: 6.04; S: 7.36.

The material possessed 8% of the bactericidal properties of Lincomycin as measured in the S. lutea test.

E-2. Methyl N- (4-pentyl-L-prolyl) -a-thiolincosaiQinide hydrochloride.

By hydrogenolysis of methyl-N- (4-pentyl-1-earboben «oxy-

0 0 9 8 2 0/1 91 6 bad original

Ii-prOlyl) -a-thiolincosaminide by the method of part £ -1 gave methyl-N - ^ - pentyl-L-prolylJ-tt-thiolincosaminide hydrochloride of melting point 212-214 °; optical rotation: + 141 ° (0.968, H ₂ O).

Analysis: Calculated for C H- ^ HgOgSCl:

C: 39.93; Hs 8.16; »: 6.13; Ss 7.02; found s C: 50.22; H: 7.96; I: 6.09; Ss 7.18.

(Corrected for 5.43 * H ₂ O).

1-3-methyl-B- (4-hexyl-L-prolyl) -a-thiolincosaminide hydrochloride.

Hydrogenolysis of methyl N- (4-hexyl-1-carbqbenzoxy-L-prolyl) <-a-thiolincosaminide by the method of Part EI gave methyl-N- (4-hexyl-L-prolyl) -u-thiolincosaminide hydrochloride, melting point 197-209 (decomposition); optical rotation [kJ-q * + 3-34 (e «0.875, H ₂ O).

Analyae: Calculated for CgQBxqNgOgSCl:

Cs 50.99; Hs 8.35; H: 5.95; found: Cs 50.32; Hs 8.07; N: 6.81.

(Corrected for $ 5.62> water).

B-4. Methyl-N- (4-oetyl-L ~ prolyl) -a-thiolineosaminide hydrochloride.

By hydrogenolysis of methyl-H- (4-octyl-1-carbo'benzoxy-L-prolyl) -a-thiolincosaminide according to the method of Part B-I obtained methyl-N- (4 <-octyl-L-prolyl) -a-thiolincosaminide-

009820/1916

⁵ < 1620G07

hydrochloride, melting point 181-200 (decomposition); optical rotation / "α J _D ί +128 (e * 0.850, HgO).

Analysis: Calculated for C ₂₂ H ^ ₂ H ₂ OgS Η01:

C: 52.94; H: 8.68; rf: 5.61; St 6.43; Found: C: 52.62; H: 8.36; N: 5.61; S: 6.36.

P. methyl - »- (4-alkyl-1-methyl-L-prolyl) -a-thiolincoeaminel hydrochloride.

. HCl. HCl

. _w 30MTL \ CV COMTL

MR

PI. Methyl-H- (4-butyl-1-methyl-L-prolyl) -a-thiolincosaminide hydrochloride.

A solution of 2.0 g of methyl I- (4-butyl-L-prolyl) -athiolincosaminid hydrochloride from part B and 2.0 g of 37% Formalin in 150 ml of methanol was subjected to 500 mg of 10 tiger palladium-carbon catalyst under 2.8 atmospheres of hydrogen pressure Shaken for 3.5 hours. after filtering off the catalyst and distilling off the solvent in vacuo partially crystalline methyl-ff- (4 ~ butyl-l-methyl-L - prolyl) -a-thiolincosaminide hydrochloride; By thin layer chromatography on silica gel using a Gtemlsoh from ethyl acetate, acetone and water (8: 4: 1) sur Sluierung and

009820/1916

BATH ORIGINAL

Potassium permanganate solution for determination was found that this product consisted primarily of two materials, namely the cis and trans epimers of methyl-H- (4-butyl-methyl-L-prolyl) -a-thiolinooeaminide hydrochloride in a ratio of about 3: 2.

Separation of the vis and trans forms by chromatography.

The methyl-N- (4-butyl-l-methyl-L-prolyl) -a-thiolincosaminide hydrochloride from Part P-I was made in a utaisch Dissolved methanol and methylene chloride (ItI), whereupon 1.5 ml of triethylamine were added. This solution became 7 g of silica gel was added and the solvent was evaporated in vacuo leaving the antibiotic on the silica gel. This Silica gel was applied to a 200 g Silica gel abandoned on top; the column was proportioned with a mixed solvent of ethyl acetate, acetone and water 8i4 * l soaked. The column was made with the same solvent developed and portions of 20 ml each were collected. Passed the thin layer chromatography described tractions 31-38 (310 ag) from essentially pure trane epimers and tractions 49-74- (32 mg) from im essentially pure cis-epiaeren. Fractions 39-4-8 consisted of a mixture of epimers that repeated by Chromatography could be further separated. Each epimer was dissolved in a few drops of dilute hydrochloric acid and that

009820/1916

Hydrochloride was precipitated by adding acetone. In this ffeise to give 50 mg of methyl N- (trans-4-butyl-lmethyl-L-prolyl) -a-thiolinco8aminid-hydroohlorid of melting point 135-157 ⁰ O and about 150 mg Kethyl-N- (cis-4 -butyl-L-lmethyl-prolylJ-a-thiolincosaminid hydrochloride softening point of 105 ° 0, further melting at 175-185 ⁰ C.

The trans-epimer recrystallized from the same solvent melted at ⁰

Analysis «Calculated for O ^

Cj 49.93; Hj 8.16; Hj 6.13; Sj 7.02; found: (corrected for 4.07 Ji HgO) j

Oj 48.81; Hi 8.54j IT: 6.49; Sj 6.67.

Recrystallization of ois-epimers was obtained an emollient at 108 ⁰ C and at about 189 ° 0 (solvated) melting cis epimer, the following analytical values ergabt

Found "(Corrected for £ 4.95 water)"

Oj 50.27; Hj 9.00; ITj 6.05; Sj 6.65;

Daa trans-Epimere was about 2.2 times as active in S. lutea-Iest as lincomycin, in the infusion broth dilution test about twice as active and in S. aureus-infected mice about. 2.5 times as active.

* Since "cis-epimers was about 1/2 to 1/3 times as active as the trans-? Epimer and had about the same activity as lincomycin"

00 9 8 2 0/1916 ^{BAD 0RIG} ' ^NAL

F-2. Methyl N- (pentyl-1-methyl-L-prolyl) -a-thiolincosaminide hydrochloride.

Reductive methylation of methyl-N- (4-pentyl-L-prolylJ-a-thiolincosaminide hydrochloride according to the procedure of Part PI gave methyl-N- (4-pentyl-1-methyl-L-prolyl) -a- thiolincoeaminide hydrochloride in the form of a mixture of cis and trans isomers, which by partition chromatography according to the method of Part FI in methyl-N- (trans-4-penty1-1-methyl-L-prolyl) -a-thiolincosaminide hydrochloride of melting point 188-191 ⁰ C and methyl-N- (cis-4-pentyl-l-methyl-L-prolyl) -a-thiolincosaminid hydrochloride of melting point 189-193 ⁰ C (135 ⁰ C sintering) was decomposed. The The trans isomer was about two to four times the bactericidal activity of lincomycin, the cis isomer one to two times the activity.

F-3. Methyl-N-i ^ -hexyl-1-methyl-L-prolylJ-a-thiolincosaminide hydrochloride.

Reductive methylation of methyl-N- (4-hexyl-L-prolyl) -a-thiolincoeaminide hydrochloride and chromatographic separation of the isomers according to the method of Part FI gave methyl-N- (trans-4-hexyl-l- me thyl-L-pr olyl) thi -α- olinc ο saminid hydrochloride of melting point 93-104- ⁰ C (decomposition) and methyl N- (eie-4-hexyl-l-ethyl-L-prolyl) -a -thiolineosaminide hydrochloride vop Sohmelx point 92-102 ⁰ C (Zereetaung). Since the trans-isomer has about two- fourfold bactericidal activity of lincomycin, the eiβ-isomer has about one, two-

009820/1916

BATH ORIGINAL

SS 162Q607

times effectiveness.

P-A-. Methyl N- (4-octyl-1-methyl-L-prolyl) -a-thiolincosaminide hydrochloride.

By reductive methylation of ^> methyl-li- (4-octyl-L-prolylj-a-thiolineosaminide hydrochloride and chromatograph! octyl-1-methyl-L-prolyl) -a-thiolincosami * nide hydrochloride with a melting point of 97-10O ⁰ C (decomposition) and methyl-N- (cie-4-octyl-1-methyl-L-prolyl) -a -thiolinooeaminide hydrochloride.

C. Methyl-H- (4-alkyl-1-ethyl-L-prolyl) -a-thiolinco-aminide hydrochloride.

.HCl

GOMTIi

MR

0-1. Methyl-N- (^ -butyl-1-ethyl-L-prolyl) -ct-thiolincoeaminide hydrochloride.

Bin e'emiech from 2.0 g of methyl-H- (4-butyl-L-prolyl) -athiolincoaaminid hydrochloride, 1.5 al of acetaldehyde and 150 mg of 10 ^ -igea palladium-carbon catalyst in 150 ml of methanol was under Shaken 2.5 atmospheres of hydrogen pressure for 5.5 hours. The catalyst was filtered off and a residue was obtained from the solution, which mainly consists of the ice

^009820/1916 BADOR, _{S, N} AL

and trans-epimers of methyl ~ N- (4-butyl-1-ethyl-L-prolyl) -athiolincosaminide hydrochloride duration.

Separation of epimers.

As described in Part F-I, the mixture of epimers from Part G-I (2 g) chromatographed on 200 g of silica gel, wherein a solvent system of ethyl acetate is used for elution, Acetone and water (8: 4: 1) used. Fractions 33-42, which thin layer fen chromatography showed to be of pure trans-epimer passed, were pooled, as were fractions 4-9-64 from pure cis epimer. Fractions 43-48 consisted of one Mixture of epimers obtained by repeated chromatography could be cleaned. Each of the epimers was dissolved in a few drops of dilute hydrochloric acid, and the crystalline hydrochloride precipitated when diluted with a large volume of ether.

The crude trans isomer (415 mg) gave 340 mg (15.4 #) of crystalline methyl-E-trans-4-butyl-1-ethyl-L-prolyl) -athiolincoeaminide hydrochloride with a melting point of 144-151 ⁰ O. The melting point was increased to 148-151 ⁰ O by recrystallization from dilute acetone. The cis epimer (645 ag) gave 300 mg (14,1 ^) of crystalline methyl-N- (cie-4-butyl-läthyl-L-prolyl) -a-thiolincoeaminid-hydiiochlorid a melting point of 135-139 ⁰ C. . By !! "crystallize from dilute acetone, the melting point was increased to 134 to 138 ⁰ C.

009820/1918

The trans-Bpimer showed in the S. lutea test the approximately 1 to 1.2 times the effectiveness of lincomycin, against gram-positive organisms did 2-4 times the effectiveness of lincomycin and against Gram-negative organisms 8 times the potency of lincomycin. In mice infected with S. aureus, this was trans-Bpimere about twice as active as lincomycin. The cis epimer possessed about half the effectiveness of the trans epimer.

G-2. Methyl-N-1-4 -pentyl-1-ethyl-L-prolylJ-a-thiolincosaminide hydrochloride.

By reductive ethylation and separation according to the process the cis and trans epimers were obtained from! eil G-I in the form of free bases and hydrochlorides. The solvated trans hydrochloride (crystallized from aqueous acetone) had a Melting point of 90-95 ° C (decomposition),

G-3. Methyl N- (4-hexyl-1-ethyl-L-prolyl) -oc-thiolineosaminide hydrochloride.

The cis and trans epimers were obtained in the form of free bases and hydrochlorides by reductive ethylation and separation according to the procedure of Part GI. The trans-hydrochloride melting at 102-121 ⁰ C (decomposition), the ice-HydrοChlorid at 93-106 ⁰ C (decomposition).

G-4-. Methyl-N- (4 ~ octyl-1-ethyl_L-prolyl) -a-thiolineosaminide hydrochloride. * ^:

By reductive ethylation and separation according to the process according to Part G-I, the cis and trans epimers were obtained

0 9 8 20/1916 BAD original

as free bases 12nd as hydrochlorides. The hydrochlorides were solid but not crystalline.

If the alkyltriphenylphosphonium bromides according to Part A of the above examples are substituted by other substituted ones Triphenyl-phosphonium-bromide with the following substituents: Methyl, ethyl, propyl, heptyl, nonyl, decyl, undecyl, dodecyl and their isomeric forms, the isomeric forms of butyl, pentyl and hexyl, Oyclopropyl, Oyclobutyl, Oyclopentyl, Cyclohexyl, Oycloheptyl, cyclocetyl, 2-0yclopropylethyl, 3-cyclohexylpropyl, Benzyl, phenethyl, 3-phenylpropyl and 1-naphthylmethyl, so one obtains the corresponding methyl-N- (cia- and trans-4- alkyl-, 4-oyclo-alkyl- and 4-aralkyl-1-earbobenzoxy-1-prolyl) -a-thiolincosaminide, Methyl-N-cis- and trans-4-alkyl-, 4-cycloalkyl- and 4-aralkyl-L-prolyl) -a-thiolincosaminidej methyl-N- (cis- and trans-4-alkyl-, 4-cycloalkyl and 4-aralkyl-1-methyl-L-propyl) -a-thiolincosaminides and the corresponding methyl egg (cis and trans-4-alkyl, 4-cycloalkyl and 4-aralkyl-1-ethyl-L-prolyl) -a-thiolincosaminides. For example, with propyltriphenylphosphonium bromide and formalin, lincomycin and Allolincomycin (cis-epimers), both active antibiotics are. If the formalin or acetaldehyde is replaced by other oxo compounds of the formula RqRqOO, for example by propionaldehyde, acetone, butyraldehyde, isobutyl methyl ketone, Benzaldehyde, phenylacetaldehyde, hydrezimtaldehyde, acetophenone, Propiophenone, butyrophenone, 3-methyl-4-phenyl-2-buta-

009820/1918

non, 2-methyl-5-phenyl-3-pentanone, 3-Gyelopentanpropionaldehyd, Gyclohexanacetaldehyd, Cyeloheptancarboxaldehyd, 2,2-Dimethylcydopropanacetaldehyd, 2,2-Bimethyloyclopropylmethylketone, Cyclopentylmethylketon, Cyclocylobutylmethylketone, Cyclobutylmethylketon corresponding methyl-N- (cis- and trans-4-alkyl-, 4-eycloalkyl- and ^ -aralkyl-1-RgRgOHj-L-prolylJ-a-thiolincosaminides, in which R _Q -RqCH denotes propyl, isopropyl, Butyl, 4-methyl-2-pentyl, benzyl, phenethyl, 3-phenylpropyl, 1-phenylethyl, 1-phenylpropyl, 1-phenylbutyl, 3-methyl-4-phenyl-2-butyl , 2-methyl-5-phenyl-3-pentyl-, 3-öyolopentylpropyl-, 2-cyclohexylethyl-, cycloheptylmethyl-, 2- (2,2-dimethylcyclopropyl) -ethyl-, 1- (2,2-dimethylcyclopropyl) - ethyl, 1-cyolopentylethyl, 1-oyclobutylethyl, cyclobutyl, cyclohexyl or 4-methylcyclohexyl radicals hyl-6- (trens-4-ethyl-L-prolylamino) -6,8-dideoxy-1-thio-D-erythro-aD-galacto-octopyranoside, methyl-6- (trans-4-ethyl-ethyl-Ir -prolylamino) -6,8-dideoxy-l-thio-D-erythro-a -D-galaotooctopyranoeid and the corresponding cis porms.

If you replace the methyl-α-thiolincοsaainid by other α-

or ß-thiolincosaainide or generally by other 6,8-dideoxy-6-amino-D-erythro and L-threo-D-galacto-octopyranose compounds of the formula III, examples of which are given below the corresponding amides of the form IV are obtained

009820/1916 bad original

and V.

Example 2 Division of allolincomycin. A. 4-Propylidene-1-carbobenzoxy-L-proline.

3.8 g of sodium hydride were heated with 75 ml Dirnethylsulfoxyd to 70-75 ⁰ C., until the reaction was completed. After cooling to 2O ⁰ C 30.8 g Propyltriphenyl bromide were added and the resulting solution was stirred for 30 minutes to ensure complete reaction. Then, a solution of 5 »2 g of 4-keto-1-carbobenzoxy-L-proline was added in 15 ml Dirnethylsulfoxyd over 15 minutes and the resulting mixture was 20 minutes at 26 ⁰ G and then 4 · hours at 7O ⁰ C stirred. The reaction mixture was cooled, 100 ml of 5% potassium bicarbonate solution and 100 ml of water were added and the mixture was filtered. The filtrate was washed twice with 150 ml of ether each time and the ether was discarded after back-extraction with bicarbonate solution. The bicarbonate solutions were combined, diluted with 200 ml of water and acidified with 4N hydrochloric acid. The acidified mixture was extracted three times with 200 ml of ether each time. The combined ether extracts were washed three times with 50 μl of saturated aqueous sodium bisulfite solution each time and then with water and dried over anhydrous sodium sulfate. After evaporation of the solvent, 5-7 g of a solid residue of 4-propylidene-1-carbobenzoxy-L-proline were obtained.

ORIGINAL 009820/1916

The residue was dissolved in 18 ml of acetonitrile and treated with 2.8 ml of dieyclohexylamine. The crystalline dicyclohexylamine salt (5.2 gj 55 # yield) melted at 154-157 After three recrystallization from acetonitrile an analytically pure sample was obtained, melting at 164-166 ⁰ C?

optical rotation Ca] ^: -8 ° (c = 0.3898, OHOL,).

Analysis: Calculated for OpgH ^ oNpO. :

0: 71.45 »H: 9.00; Ni 5.95i found: Oi71.77J Hi 9.39} N: 5.1.

8 g (17 millimoles) of the dicyclohexylamine salt were shaken with excess 1.5N sodium hydroxide solution and ither, until complete dissolution occurred. The layers were separated and each was back washed. The watery alkaline phase was combined with the Wasohlösungen and with 4-11 hydrochloric acid acidified. The mixture was extracted with ether and the ether extracts were combined and evaporated to give 4.8 g (97.8 #) of 4-propylidene-1-carbobenzoxy-L-proline in the form of an oil received.

B. Methyl N- (4-propylidene-1-carbobenzoxy-L-prolyl) -athiolincosaminide.

+ UTL COOH

COMTL

00 98 20/1916

BATH ORIGINAL

1.08 ml of isobutyl chloroformate in 1 ml were added to a solution of 2.25 g of 4-propylidene-1-carbobenzoxy-L-proline from Part A and 1.40 ml of triethylamine in 80 ml of distilled ^{acetonitrile cooled to 0 ° C.} Acetonitrile added. The mixture was ^{stirred at 0} ° C. (+ 5 ^° C.) for 15 minutes. A solution of 2.92 g of methyl α-thiolincoaaminide (MTL) in 100 ml of water was then added rapidly. The resulting solution was ^{stirred at O 0} O for one hour, the cooling bath was removed and stirring was continued for an additional hour. The acetonitrile was distilled off in vacuo, a partially crystalline residue remaining. The mixture was cooled to 1O ⁰ C, filtered and the product was dried in vacuo at 55 ⁰ C to yield 2.3 g of Crystallizing methyl-N- (4-propylidene-l-carbobenzoxy-L-prolyl) -a- thioliiicosaminid received ⁰ C of melting point 178-186, Huh recrystallized twice from ethyl acetate damp an analytical sample with a melting point 180-187 ⁰ C was obtained; optical rotation [a] _D : + 137 ° C (MeOH), c = 0.92).
Analyzes Calculated for CgcH ^ gHgOgS:

Os 57.23} H: 6.92; N: 5.32; Si 6.11; found? C: 57.24; Hs 7.22; Hs 5.18; Ss 6.16.

C. Methyl 4- (4-propyl-L-prolyl) -α-thiolincosaminide hydrochloride.

.HCl

COMSL

0 9 8 2 0/1916 BAD original

A solution of 100 mg of methyl-N- (4-propylidene-1-Csxbobenzo ^ -L-prolylJ-a-thiolincosaininide, obtained according to Part O, in 50 ml of methanol was over 100 mg of 7 # platinum Dowex- 1 catalyst was shaken at 2 * 8 atmospheres hydrogen pressure for three hours. Then 100 mg of 10 # palladium-carbon catalyst were added and the reaction mixture was shaken under 2.8 atmospheres for a further three hours. The catalyst was filtered off and the solvent was distilled off in vacuo. the residue was dissolved in 0.1 ml of 0.5 -Salzsäure η. When diluted with 15 ml acetone, the _f 0 methyl-N- fell (4-propyl-L-prolyl) -a-thiolincosaminid hydrochloride of which was filtered off, the crystalline product weighed after drying at 55 ⁰ O in vacuo 20 mg;.! melting point 181-188 ⁰ C. the product contained about four parts per part cis isomer trans-Ieomer..

D. Methyl-Η- (4-propyl-1-methyl-L-prolyl) -oc-thiolincosaminide hydrochloride.

^H .HCl ■ .HCl

COMTL J COMTL

A solution of 100 mg of methyl-H- (4-.propyl-L-prolyl) -athiolincosaainide hydrochloride, obtained according to Part C and 0.2 ml Formalin in 50 al of methanol was over 100 mg of 10 tigern palladium-carbon catalyst at 2.8 atmospheres of hydrogen pressure for 5 hours long shaken. The catalyst was filtered off and that

ÖAD 009820/1916

Solvent was removed in vacuo. The residue showed two spots on thin-layer chromatography, one of which is lincomycin hydrochloride and the other is allolincomycin hydrochloride corresponded.

E.! Separation of the vis and trans forms by chromatography.

One gram of methyl N- (4-propyl-1-methyl-L-prolyl) -a-thiolincoaaminide hydrochloride, prepared according to part D, was in 15-20 ml of methylene chloride containing 0.5 ml of triethylamine, solved; 2 g of silica gel was added. The solvent was then distilled off in vacuo, a residue of the Antibiotic was left on the free flowing silica gel. 100 g of silica gel was slurried with 80% aqueous acetone and placed in a chromatography column. The solvent was drained to the level of the old man. That Antibiotic silica gel was placed on top of the column, followed by a layer of sand. The pillar was eluted with 80% aqueous acetone, collecting tractions of 20 ml each. Every traction was turned into Evaporated dry and on silica gel by thin layer chromatography investigated using 80 # aqueous acetone for Elution was used. The antibiotic is found by spraying on alkaline permanganate solution. the Fractions containing the desired product were combined, evaporated to dryness and the antibiotic crystallized as the hydrochloride when dissolved in excess dilute salt

00982071916

BATH ORIGINAL

CS

acid and diluting with acetone. Pas trans isomer was eluted first, followed by a mixture of cis and trans isomer and finally the pure cis isomer. The mixture can be chromatographed again as described above. Pas cis isomer, allolincomycin hydrochloride, had a melting point of 14-7-15O ⁰ O; optical enhancement CaJ ₀ : + 110 (HgO).

Analysis: Calculated for O ₁₈ H- ^ N ₂ OgS. HOl:

0: 48.80; H: 7.96? Ns 6.32; found: (Corrected for 9 »47 $> HgO);

C: 49.15? H: 7.80; N: 6.39.

Replaces the propyltriphenylphosphonium bromide in part A of the above example with other substituted triphenylphosphonium bromides with, for example, one of the following substituents: butyl, pentyl, hexyl, heptyl, octyl, nonyl, pecyl, undecyl, Podecyl or its isomeric forms, methyl, ethyl, isopropyl, Cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, Oycloheptyl, Oyclooctyl, 2-Cyclopropyläthyl, 3-Oyclohexylpro-- pyl, Benzyl, phenethyl, 3-phenylpropyl or a-naphthylmethyl, the corresponding methyl-N- (cis- and trans-4-alkylidene, cycloalkylidene and aralkylidene-l-carbobenzoxy-L-prolyl) -α-thiolincοsaminide, Methyl-N- (cis- and trans-4-alkyl-, cycloalkyl- and aralkyl-L-prolyl) -a-thiolincosaminide and the corresponding N- (cis- and trans-4-alkyl-, cycloalkyl- and aralkyli-methyl-L-prolylJ-a-thiolincosaminides obtain. Replace one

009820/1916

BAD ORIG'NAL

the formalin in Part D by other Eetaldones of the formula RgRgCO, for example by acetaldehyde, propionaldehyde, acetone, butyraldehyde, isobutyl methyl ketone (4 ~ methyl-2-pentanone), Benzaldehyde, phenylacetaldehyde, hydrocinnamaldehyde, Acetophenone, propiophenone, butyrophenone, 3-methyl-4-phenyl-2-butanone, 2-methyl-5-phenyl-3-pentanone, 3-cyclopentane propionaldehyde, Cyclohexane acetaldehyde, cycloheptane carboxaldehyde, 2,2-dimethylcyclopropane acetaldehyde, 2,2-dimethylcyclopropyl methyl ketone, Cyclopentyl methyl ketone, glyclobutyl methyl ketone, oyclobutanone, Cyclohexanone and 4-methylcyclohexanone, the corresponding methyl-N- (cis- and trans-4-alkyl-, 4-cycloalkyl- and ^ -aralkyl-l-RgRftCH-L-prolylJ-a-thiolincosaminide, in which RqRqCH denotes ethyl, propyl, isopropyl, butyl, 4-methyl-2-pentyl, benzyl, phenethyl, 3-phenylpropyl, 1-phenylethyl, 1-phenylpropyl-, 1-phenylbutyl-, 3-methyl-4-phenyl-2-butyl-, 2-methyl-5-phenyl-3-pentyl, 3-cyclopentylpropyl-, 2-cyclohexylethyl, cycloheptylmethyl, 2- (2,2-dimethylcyclopropyl) ethyl, 1- (2,2-dimethylcyclopropyl) ethyl, l-Cyclopentylethyl, 1-cyclobutylethyl, cyclobutyl, cyclohexyl or 4-methylcyclohexyl radical.

Replace the methyl-a-thiolincosaminide with others α- or ß-thiolincosaainide or in general by other 6,8-dideoxy-e-amino-D-erythro-D-galacto-octopyranose compounds of the formula III, for which examples are given below, corresponding amides of the formulas IV and V are obtained.

009820/1915

BATH ORIGINAL

Example 5: Ethyl-α-thiolincοsaminid.

Two grams of lincomycin C hydrochloride were in 50 ml Dissolved water and the pH of the solution was adjusted to 9-5 by adding an ion exchange resin in the hydroxy form (The anion exchange resin was obtained by Chloromethylation of polystyrene, optionally crosslinked with divinylbenzene, and quaternization with trimethylamine or ' Dimethylethanolamine; For manufacture see Kunin, Ion Exchange Resins, 2nd Edition, (1958), John Wiley and Sons, Inc., pp. 88, 97). The alkaline solution was then freeze-drying subjected to a residue obtained in 50 ml 93-100 # hydrazine hydrate was dissolved, followed by 24 hours boiled on reflux for a long time. The solution was then evaporated to dryness in vacuo and the Hüekestand was three times with each 10 ml of acetonitrile triturated. The soluble material was collected and dried, yield 900 mg. A® Merasg of 600 mg of the dried insoluble material in 4 ml of dimethylformamide (the solution was made with heating) was clarified by filtration and the piltrate was left for 4 hours at room temperature held. The precipitating crystalline ethyl-a-thiolincoaaminid was filtered off, washed with ether and dried; Yield 500 mg.

Ethyl-α-thiolincosaminid has the following physical and chemical properties "Melting point: 191-195 ⁰ C; optical rotation Ca] ² S: + 258 ⁰ C (c = 0.76 in water) pKa ¹ = 7.17.

009820/1916

BATH ORIGINAL

Ib ^ UbU / 69

Analysis: Calculated for

0: 44.93; H: 7.92; N: 5.24} S: 11.99; 0: 29.92; found:

0: 44.09; H: 7.91? N: 5.24; Ss 11.32.

Pas Lincomycin C hydrochloride was prepared as follows: Fermentation:

A slanted culture medium with Streptomyces lincolnensis var. lincolnensis, NRRL 2936, was used to inoculate 500 ml Erlenmeyer flasks, each 100 ml of one The medium contained the following components:

Yeastolao 10 g Glucose monohydrate 10 g NZ-amine B ⁺⁺ 5 g Tap water to the Padding on 1 liter

⁺ Yeastolac is a protein hydrolyzate made from yeast cells.

⁺⁺ NZ-Amin B is enzymatically degraded casein from Sheffield.

The pre-sterilization pH was 7.3. It was grown for two days at 28 ^° C. on a Gump shaker at 250 rpm.

5 i "inoculum of the above medium (5 ml) were respectively 30,500 al Erlenmeyer flask entered, each containing the following fermentation medium:

0 0 9820/1916

BATH ORIGINAL

69 Grluco semonohydrate 15 g 1620607 strength 40 g Holassen 20 g Wilson's Peptones Liquor Ho. 159 ⁺ 10 g Gorn Steep Liquor 20 g Calcium carbonate 8 g Lard oil 0.5 ml Tap water to the Padding on 1 liter

⁺ Wilsoh's Peptone Liquor No. 159 is a preparation made from enzymatically hydrolyzed proteins of animal origin.

At the time of inoculation, DL-ethionine was in added to a concentration of 2 mg per ml.

After fermentation for 4 days at 28 ^° C., the shake flasks were worked up on a Gump shaking machine at 250 rpm. The content was 200 meg per ml, determined according to the S. lutea-Iest described below. There was about 20 g of solids per liter in the entire fermentation broth.

Cleaning:

The entire fermentation broth (235 l) from a DL-lthionone permentation was filtered at the work-up pH, if necessary with the use of a piltering aid

0 098 20/1916 bad ORiGiNAt

Mycelial cake was washed with water and discarded. That The filtrate and washing solutions (275 1) were used for 45 min 12.5 kg of activated carbon and 2.5 kg of diatomaceous earth were stirred. That Mixture was filtered and the filtrate was discarded. The charcoal cake was washed with 60 l of water and the washing solution was discarded. Then 70 l of 20 # aqueous Acetone washed, which was also discarded. Then was eluted twice with 100 liters of 90 # aqueous acetone each time. The eluates (215 l) were combined and concentrated to 18 l. The concentrate was diluted with 50 # aqueous sodium hydroxide solution adjusted to a pH of 10.0 and extracted three times with 20 l of methylene chloride each time. The methylene chloride extracts (60 1) were combined and concentrated to give an oily Preparation (7.14 x g) containing lincomycin and lincomycin 0 in equal amounts, both in the form of the free base. The preparation was dissolved in 200 ml of methylene chloride. The solution was clarified by filtration and then concentrated to dryness in vacuo. The residue was dissolved in 100 ml of In-Methano-Iisehem Dissolved hydrogen chloride, whereupon the methanol solution was mixed with 3 »2 1 ether while stirring. The one that fails colorless crude lincomycin hydrochloride and lincomycin 0-hydrochloride was filtered off and dried; Yield 7.14 x g with a content of 94-0 meg per mg against Sarcina lutea (the Test against Sarcina lutea is carried out on agar, which is adjusted to a pH of 6-8 with pH 7.0 phosphate buffer CO, 1 M]

009820/19 16 ^{ßAD 0RiG1} NAt

is buffered. You unit volume CO, 08 al] of that to be tested The solution containing the material is placed on a 12.7 ml test plate, which is then transferred to a test organism Agar plate is applied). Thin layer chromatography indicated the presence of lincomyoin hydrochloride and lincomycin G-hydrochloride in approximately equal amounts.

Crude lincomycin C hydrochloride (7.0 g) was added in 20 ml Water and 20 ml of butanol dissolved, whereupon the pH of the solution adjusted to 4.2 with in-salic acid; the solution was in a (Jegenstroa distribution apparatus for 1000 transitions. Thin-layer chromatographic analysis showed that the Fractions in tubes 135-190 contained lincomycin 0. These fractions were combined and the solution became one concentrated aqueous solution and then subjected to freeze-drying, whereby 2.44 g of lincomycin C hydrochloride were obtained, the against Sarcina lutea showed 1400 meg per mg. 500 mg of this pre. parates were dissolved in 2 ml of water, 1 ml of methanol and 100 μl of acetone. The solution was filtered and the filtrate became bit Ether mixed until crystals appeared. Bann was left to stand for 1 hour at room temperature. Crystalline Lincomycin C-hydrochloride (cube) was made from the supernatant solution separated by decantation. The crystals were recrystallized from 1 ml of water, 1 ml of methanol, 80 ml of acetone and 20 ml of ether, 250 mg of the cubic crystalline lincomycin C hydrochloride being obtained. The decanted solution (

_: 009820/1916

BATH ORIGINAL

same above) was left to stand ^{at 5 ° C. for 4 hours.} Acicular-crystalline Lincomycin C-hydrοChlorid precipitated out, which was filtered off and dried. The yield of the latter product was 150 mg, mp 151-157 ⁰ O.

If the procedure of Example 1 is followed, but replacing the methyl-a-thiolincosaminide with ethyl-ethiolincosaminide, compounds of the formulas IV and V are obtained in which R, R- and Z have the above meaning and Y the grouping -SCELCH * in α- configuration. For example, using an alkyltriphenylphosphonium bromide and formalin or acetaldehyde, the antibiotically active ethyl-6- (trans-substit.-L-prolylamino) -6,8-dideoxy-1-thio-D-erythro-aD-galacto-octopyranoside is obtained may represent, and the corresponding cis-forms, with the Subatituenten 4-alkyl, 4-alkyl-l-methyl and 4-alkyl-l-ethyl, in which the alkyl group is ethyl, propyl, butyl, pentyl, H _e xyl or isomers thereof .

Example 4; Alkyl-ß-thiolincosaminides.

S-alkyl

XXXV

009820/1916

A. Methyl-N-aeetyl-a-thiolincosaminide.

Five grams of methyl α-thiolincosaminide (about 0.02 mol) were suspended in 50 ml of methanol with stirring and treated with 4.04 g (about 0.04 mol) of acetic anhydride. The starting material dissolved almost completely and then a solid was encountered. After 18 hours at room temperature (approx. 25 ⁰ O) the solid was filtered off, washed with methanol and dried in a vacuum drying cabinet at 50 ° O and 15 mm pressure. 4.58 g ( $ 79) of a crystalline product with a melting point of 242-245 ⁰ O were obtained. This product was recrystallized from absolute methanol, colorless needles of methyl-N-aetyl-ct-thiodine being obtained
25th

lincosaminide with a melting point of 243-245 ° ö; optical Rotation [a3 J. + 265 0. (c = 0.7374, water).

Analyzes Calculated for ⁰ I ₁ H ₂ I ^ ⁰ O ⁸ *

C: 44.72; H: 7.17i Ns 4.74; S: 10.85? found 44.87; H: 7.10; N: 4.65 *, S: 10.99.

B. Mixture of e-Acetamido-e ^ -dideoxy-D-erythro-a- and

ß-D-galacto-octopyranonsen (N-acetyl-α- and ß-lincosamines).

A solution of 50 g of methyl N-acetyl-α-thiolincοsaminid, prepared according to Part A, in 1500 ml of water at 40 ⁰ O was stirred with a magnetic stirrer and with a solution of 70 g of mercury (II) chloride in 1500 ml of water of 40 ⁰ C treated. A white precipitate formed immediately. The reaction was continued for three days with occasional warming to 40 ^{° C.} Thin-layer chromatography thus

009820/1916 BAD original

then demonstrated the lack of starting material. The precipitate of mercury chloride mercaptide (GlHgSMe) was filtered off. The colorless piltrate was stirred together with the washing solutions of the precipitate (aqueous) with a magnetic stirrer at room temperature and excess mercury chloride was removed by adding pyridine in portions and precipitating the insoluble mercury chloride-pyridine complex. After the mixture had been left to ^{stand for three hours in the refrigerator at 0} ° C., the pesticide was filtered off, the residue was washed well with cold water and the filtrate and washing solutions were stirred with a little silver carbonate until the solution was neutral to pH paper. Then was filtered through a "Millipore ^w filter (Millipore Filter Corporation, Bedford, Massachusetts), filtered, the filter was thoroughly washed with water and the washings were added to the colorless FiItrat excess silver ions were by saturating the solution with hydrogen sulfide, and filtering off the silver sulfides. The silver sulfide was washed with water and the washing solutions were added to the filtrate - the solution was lyophilized to give a colorless amorphous solid which was composed of a mixture of e-acetamido-öfe-dideoxy-D-erythro-a- and ß -D-galactooctopyranose (N-acetyl-a-lincosamine and N-acetyl-ß-lincosamine).

The raw semi-sch from N-Acetyl-cc-lincosaain and N-Acetyl-

009820/1916

ß-lincosamine, obtained according to Part B, was slurried in 400 ml of pyridine and 200 ml of acetic anhydride and stirred with a magnetic stirrer overnight at room temperature. The resulting colorless solution was concentrated on a drum evaporator at 40 ^° C. and 1 mm Hg to a pale yellow syrup, which was dissolved in a mixture of water and chloroform. The aqueous layer was extracted with chloroform and the chloroform extracts were combined, washed with dilute sulfuric acid (2-normrl), then washed twice with water and with saturated aqueous sodium bicarbonate solution, then again with water until neutral and finally dried over anhydrous sodium sulfate. The resulting Gtoloroformextrakt was then evaporated on a Xrommelverdampfer at 5S ⁰ C and 15 mm Hg to give a colorless solid which Wurd dissolved in hot ethyl acetate @ _e This Äthylaeetatlösung was Skellysolve B-hexane was added until the onset of crystallization. The solid obtained in this way was filtered and recrystallized twice from ethyl acetate-Skellysolve B-hexane, N-acetyl-1,2.3f4 _t 7-penta-0-acetyl-ß-lincosamine (9t27 g) with a melting point of 227-230 ^{Received 0} C; optical rotation [<x] | ⁵ : + 33 ⁰ C (c = 0.832, chloroform).

Analysis: Calculated for Cg ₀ H ₂ QNO ₁₂ :

C: 50.52; H: 6.15 » ^H * 2.95; found: C: 50.40; Ht 6.42; N: 3.04.

BATH ORIGINAL 009820/1916

The ethyl acetate-Skellysolve B-hexane filtrate from the first crystallization was left to stand at room temperature, with colorless prismatic needles made of N-aoetyl-1,2,3 »4,7-penta-O-acetyl-a-lincosamine with a melting point of 169 -172 ⁰ C. The melt solidified on cooling in the form of hexagonal platelets, which then melted again ^{at 237-238 0 G;} after recrystallization from methyl acetate-Skellysolve B to give hexagonal platelets of melting point 240 to 240.5 ⁰ O; optical rotation Ioclj " ⁵ : + 132 ° (c« 0.9842, chloroform).

Analysis: Calculated for CpgHpg ^

C; 50.52; H: 6.15; N: 2.95; found: G: 50.62; H: 6.08; Nt 3.02.

The N-acetyl-1,2,3,4,7-penta-0-acetyl-α- and ß-lincosamines can also be separated by Gregenstrom distribution using a system of water: acetone: methyl ethyl ketone: cyclohexane in a volume ratio of 3s5s4 * 4 . With 900 transitions about 50 % of each of the two isomers could be isolated in pure form; the distribution coefficient is 0.75 for the oc anomer and 0.66 for the ß anomer.

D. 6-Acetylamino-2,3,4,7-tetra-O-acetyl-1-bromo-1,6,8-trideoxy-D-erythro-α-D-galacto-octopyranose (N-acetyl-2,3f4,7-tetra-O-acetyl-1-bromo-l-deoxy-a-lincosamine) .

AcO -j— AcNH -l—

^ACO '-0 XXXVI

OAc

Br

OAc

009820/1916

BATH

Two grams of N-acetyl-1,2,3 »4,7-penta-O-acetyl-ß-lineosamine, prepared according to Part O, were mixed with a saturated solution of anhydrous hydrogen bromide in 5 ml of acetic acid at room temperature (approx. 25 ⁰ C) for about 3 hours with a magnetic stirrer. All solids dissolved within an hour. The pale yellow viscous solution was diluted with 50 ml of chloroform, poured onto ice and stirred for 10 minutes. The chloroform layer was separated, the aqueous solution was carefully extracted with chloroform and the combined chloroform extracts were washed until the aqueous washing solution was neutral to Congo paper; then it was dried over anhydrous sodium sulfate. The chloroform was ^{evaporated in a drum evaporator at 35 °} C. and 15 mm Hg, an almost colorless, amorphous solid being obtained. After three times of recrystallization of the solid from chloroform-Skellysolve B N-Acetyl-2,3> 4,7-tetra-0-acetyl-la-bromo-l-deoxylincosamin was obtained as colorless prismatic needles, melting point 188-189 ⁰ C. ; optical rotation Ca] ² ^ _s + 231 ⁰ C (c = 0.8132, chloroform).

Analysis: Calculated for C ^ gHggBrNO- ^:

C: 43.56; H: 5.28; N: 2.82; Br: 16.10; Found: C: 43.68; H: 5.39; N: 2.88; Br: 17.22.

E. Methyl-N-acetyl-2,3,4,7-tetra-0-acetyl-β-thiolincosaminide.

Two grams of N-acetyl-2,3,4,7-tetra-O-acetyl-la-bromo-1-deoxylincosamine, prepared according to part D, were in 25 ml

009820/1916 BADORlGtNAL

Acetone, which had previously been dried over potassium carbonate, dissolved; 350 mg of thiourea were added to the solution. After brief warming on the steam bath to dissolve the solids, the colorless reaction mixture was allowed to stand overnight at room temperature. Then a solution of 680 mg of potassium carbonate and 860 mg of sodium bisulfate in 10 ml of water was added, after which 900 mg (0.40 ml) of methyl iodide were added. The mixture was stored in a sealed bottle and shaken at room temperature for 3 hours. It was then carefully extracted with chloroform, the combined extracts were washed twice with water, dried over anhydrous sodium sulphate and ^{freed from the solvent on a drum evaporator at 40 °} C. and 15 mm Hg. In this way, 1.38 g of a colorless, amorphous solid was obtained which, on thin layer chromatography, was found to consist of a single compound. The solid was dissolved in hot ethyl acetate and then diluted with Skellysolve B hexane; there was obtained 880 mg of colorless flakes of melting point 268-272 ⁰ C. After recrystallization from the same solvent was added methyl-N-acetyl-2,3 '4,7-tetra-0-acetyl-ßthiolinoosaminid of melting point 272-273 ⁰ C obtained; optical rotation [<x3 ^: + 31 ° (c * 0.6800, chloroform).

Analysis: Calculated for

C: 29.22; H: 6.31; Ns 3.02; Ss 6.92; Found: C: 49.15; H: 6.23; N: 3.00; S: 6.41.

009820/1916 ^ AD original

I ¹ . Methyl-N-acetyl-ß-thiolinco saminide.

• *

10 ml of methanol were saturated WLT dry ammonia at 0-5 ⁰ O ge. At Raisiateaperatiir 1 g of methyl-N-acetyl-2, 5 »4-t 7-tetra-O-aoetyl-ß-thiolincoaamisiid was given to this solution. The Semisch was 3 Stimöea long to stand at Hauatemperatui ', and thereafter, on a TrommelveräaEpf it at 40 ⁰ C and 15 evaporated to dryness na Hg; the residue obtained in this way was crystallized three times from ethanol _9, with maxi receiving the methyl-N-acetyl-ß-thioliaeosamic acid.

0. Methyl-S-thiolincosaaiiSide.

A solution τοπ 1 ι
Boiled in 10 ml of hydrazine hydrate wmä® for 24 hours on the river. Excess hydrazine was removed in a dry nitrogen stream on the agfb & d eat. The crystalline residue was recrystallized from 5 ml of water, the crystals were collected, washed with cold water and dried in vacuo to give the above compound «

If you replace the methyl iodide from the above part £ by ethyl, Propyl, butyl, pentyl, hexyl, heptyl, octyl, honyl, Decyl, undecyl or bodecyl iodide or corresponding isomers, so the corresponding alkyl-H-acetyl-ß-thiolincoeaminid- and alkyl-β-thiolincosaminide.

Following the procedure of Example 1, compounds of formula

009820/19 16

Σ
ι \ ι HO - OH, \ Λ HH - -4 η ι O ^Η ° κ OH

XXXVII

OH

in which X is «R-, or Z according to the above meaning» and R is the has the above meaning, received *

If the methyl;) odide in the above part E is replaced by Z- hydroxyethyl iodide, a-hydroxyethyl-N-acetyl-a ^ -4,7-'tetra-O-acetyl-ß-thiolincosaminide, 2-hydroxyethyl-H is obtained -acetylß-thiolineosaminide and 2-hydroxyethyl-ß-thiolineosaminide. The dead-speaking 2-alkoxyethyl compounds are prepared using 2-alkoxyethyl iodides. The alkyl group in the alkoxy radicals can be, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl or their isomers.

Following the procedure of Example 1, compounds of the formula

009820/1916

BATH ORIGrNAL

GH,

HO HH HO

O S-OH

XXOTII

obtained, in which X = R ₁ or Z is as defined above and H has the meaning given above, and in which IL is hydrogen or alkyl.

Example 5t 6-Amino-1,6,8-trideoxy-D-erythro-D-galacto-octopyranose (1-deoxylincosamine).

OH,

HO H ₂ N

HO

OH

OH

YYYTY

BAD ORIGtNAL

009820/1916

Λ. Methyl-N-acetyl-3> 4> 0-isopropylidene-e-thlolinco8aninld.

Slue suspension of 5.3 g of fine powdery methyl 1-N-acetyl-ß-thiolincoeaminide was stirred for 30 minutes at room temperature with ^ 00 ml of acetone and 5 ml of concentrated sulfuric acid. After 30 minutes a further 5 ml of concentr. Sulfuric acid was added and the solution was stirred for an additional half hour at tree temperature. Then 150 g of barium carbonate in 100 ml of water were added and the mixture was stirred until the mixture was neutral. Barium sulfate and excess carbonate were filtered off and the precipitate was washed with acetone. Piltrate and washing solution were combined and evaporated to dryness ^{in vacuo at 40 0 O.} The residue was treated with acetone ether (10: 1) and insoluble material was filtered off. The piltrate was evaporated to dryness and the residue was dissolved in 100 ml of ethanol. After removing the alcohol by heating in vacuo, a gummy material remained which was dissolved in 20 ml of warm water containing a little sodium carbonate. After small amounts of insoluble matter had been filtered off, the piltrate was cooled. After standing in the refrigerator for 4 hours

0098 20/1916

the crystals that had formed collected, washed with cold water and dried in vacuo; this gave methyl-H-acetyl-3,4-O-isopropylidene-β-thiolincosaminide.

B. I-Acetyl-3,4-0-isopropylidene-1-deoxylinoosaiine. '

5 g of methyl-H-acetyl ^ f * -0-ieopropylidene-fl-. thiolincosaminide were refluxed with 38 ml of loose Raney nickel in 150 ml of ethanol for 7 hours. The mixture was then filtered and the catalyst was washed with a total of 4-00 ml of boiling ethanol. Pil stepped and wash solutions were combined and evaporated to dryness to give a partially crystalline residue. This was purified by countercurrent distribution in a system of butanol / water and in this way Έ- acetyl-3,4-0-isopropylidene-1-deoxy-linoosamine was obtained.

0. 1-Deoxylincosamine.,.

1 g of 1-acetyl-3,4-0-ieopropylidene-1-deoxylincosamine in 5 ml of hydrasin hydrate (98-100 jt-ig) was for 21 hours refluxed, excess β hydrazine hydrate was

00 9820/1916. -. BADORIQiNAL

then distilled off in vacuo. The residue was stirred in old acetonitrile, whereupon it was again evaporated. Then 5 ml of water were added and the pH was adjusted to 2 using hydrochloric acid. After 3-5 hours at 26 ^° C., the solution was diluted 10 μl of water and old excess silver carbonate was added. It was then filtered and the piltrate was lyophilized. The resulting residue was crystallized from water to obtain 1-deoxylincosaaine.

Following the procedure of Example 1, compounds of the following formula can be obtained:

CH,

HO NH

XL

in which X «R or Z is old of the above meaning, and R has the above meaning.

009820/1916

Example 6; 2-hydroxyethyl-6-amino-7-0-ethyl-

6,8-dideoxy-1-thio-D-erythro-a-D-galacto-octopyranoside (2-hydroxyethyl «a-thiocelestosaminide).

XLI

SOH ₂ CH ₂ OH

A. 2-Hydroxyethyl-a-thioceleatosaminide hydraain solvate.

A mixture of 5 g (0.0094 hol) celesticetin (Example 3 of US Pat. No. 2,928,844) and 25 ml (excess) hydrazine hydrate was refluxed for 21 hours. Excess hydrazine was removed by distillation in vacuo and the residue was recrystallized from 35 ml of absolute ethanol. There was obtained 1.2 g of 2-hydroxyethyl-a-thiocelestosaminidhydrazin solvate in the form of white crystals of melting point 98-108 ⁰ C. After recrystallization from absolute ethanol, 0.65 g of product having the optical rotation

BAD 009820/1916

[aj ² fj ■ ϊ + 243 ° (C * 0.8 water). The IE spectrum in Nujol showed the following bands 3400, 1630, 1600, 1460, 1450 (Sch), 1305, 1275, 1260, 1200, 1115, 1085, 1055, 1010, 978, 950, 925, 910, 873, 800- 820, 705, 690 and 680 approx. The equivalent weight was 161 with two basic groups of bit pKa values in the range of 7.5.

Analysis: Calculated for

Ct 40.11; Hs 8.26; H: 12.76; S: 9.74; found: C: 40.15; H: 8.04; Hs 11.69; Ss 9.56.

B. 2-Hydroxyethyl-a-thioceleetoBaminid-hydrazine-solvate.

10 g Desalicetin (comp. Example 1 of USPatentschrift 2,851,463) were dissolved in 100 ml of hydrazine hydrate and refluxed in an oil bath at 165 ⁰ C for 18 hours under reflux. The almost colorless solution was evaporated to dryness on the steam bath, first at 15 mm and then at 1 mm, leaving a solid residue which was triturated with acetonitrile, filtered and washed with the same solvent. Crystallization from ethanol gave 2.64 g of 2-hydroxyethyl-a-thiocelestoeaminide hydrazine solvate in the form of colorless hadels. When recrystallizing from the same

BATH ORIGINAL 009820/1916

Solvent obtained crystals from 2-Hydroxyäthyl ox thiocelestosamihid-hydrazine solvate with an Äquivalentgewi ent tone 168 and two basic groups with pKa values in the range of 7.5; optical rotation ^ 5, ₊ 248 ° (c »lift 95 * -igem ethanol).

Analysis » Calculated for C ₁₁ H ₂ ^ HOgS.

■ Ct 40.11; H * 8.26; Hi 12.76; Si 9.74; found C * 40.27; H: 7.95; H: $ 11.63 S: 9.80.

0. 2-Hydroxyethyl-a-thiocelestoeaminide.

A solution of 2 g of 2-hydroxyethyl-a-thiocelestosaminide hydrazine solvate, prepared according to Part A or B, in 30 ml of dimethylformamide was concentrated to a volume of about 10 ml and then diluted with 10 ml of dimethylformamide. As soon as the cloud began to appear, Xther was added and then 500 mg of 2-hydroxyethyl-ethiocelestosaminide separated out. The crystalline 2-Hydroxyäthylox thioceleetosaminid showed an optical rotation of [a] | ³ : + 262 ° (c - 1 in water); IE spectrum: 3400 (Sch), 3250, 1600, 1400, 1325, 1310, 1290, 1240, 1195, 1160, 1150 (Sch), 1110, 1100, 1075, 1045, 1038, 1005, 980, 920, 895, 862, 825, 796, 740, 711 and 690 cm "¹; equivalent weight 297, pKa» 7.2.

00 9820/1916

Analysis: Calculated for

G: 44.43; H: 7.80; H: 4.71; S: 10.78; found »C: 44.20; H: 7.78; N: 4.97 »Ss 10.68.

According to the method 70η Example 1 know compounds

the formula

XLII

S-CHgOHgOH

OH

in which X = R., or Z geaäß of the above meaning, and in which also R has the above meaning.

Example 7 " 7-0-methyl-1-deoxylincosamine.

OH,

H ₂ N HO.

ZLIII

009820/1916

BATH ORIGINAL

A. 2-Hydroxyethyl-N-acetyl-3,4-0-isopropylidene-α-thiooelestosaminide.

To a solution of 14 g (0.047 mol) of 2-hydroxyethyl-a-thioeelestosaminide 14 ml of acetic anhydride were added to 150 ml of ethanol. The reaction mixture was stirred for half an hour, cooled over Baoht and then evaporated to dryness in vacuo. The residue from ^ -Hydroxyäthyl-N-acetyl-ct-thiocelestosaminid was in Slurried ether and dried under reduced pressure.

The dry material obtained in this way was then dissolved in 1500 ml of acetone and concentrated while stirring with 15 ml. Sulfuric acid decomposes. The mixture was stirred for about 2 hours and then by the addition of dry ammonia neutralized. It was then filtered and the piltrate was evaporated to dryness, with 2-hydroxyethyl-] i-acetyl-3 »4-O-isopropylidene-a-thiocelestosaminide when oil was left behind.

B. -1 JEr-Acetyl-3,4-0-isopropylidene-7-0-methyl-l-deoxy-

lincosamine.

The Ul prepared according to Part A was in 500 ml Dissolve ethanol and add 150 ml of loosely packed Raney-Hickel added to ethanol; the mixture was then 10 hours.

0 09820/1916

refluxed for a long time, then filtered, the catalyst washed with 1 liter of boiling ethanol and the combined filtrate and washing solutions were evaporated to dryness. An UI was obtained that (with 500 transitions) in the system 1-butanol: water was distributed. Its main fraction, K = 0.82, was obtained on evaporation of the tubes 200-250 and gave 4.6 g (33 #) N-acetyl ^^ O-ieopropylidene ^ -O-methyl-1-deoxylincosamine »which crystallized on drying.

The melting point was 198-205 ° C; optical rotation [<x] | ⁵ t + 71 ° (c = 1.50 i »ethanol).

Analysis: Calculated for

C: 55.4-3; Hj 8.31; F: 4.62j methoxyl: 10.63; Found C: 55.03; H: 8.28; H: 4.70; Methoxyl: 10.43.

1.16 g of potassium metal were dissolved in 100 ml of tert-butyl alcohol. The solvent was removed by distillation (at normal pressure) as far as possible and then through Vacuum distillation removed at 15 mm Hg. The dry one solid residue, 100 ml of dry benzene were added, which was then distilled off and a fine powder remained. The potassium tert-butoxide thus obtained was

BATH ORIGINAL 009820/1916

200 ml of dry benzene was added and the mixture was stirred with a magnetic stirrer at room temperature until an opalescent solution was obtained. This solution was mixed with 5 g of H-acetyl-Jf ^ -O-isopropylidene-1-deoxylinoosamine, prepared according to Part B of Example 5, and the mixture was then stirred at room temperature overnight. To the resulting mixture, 42.4 g (18.6 ml) of methyl iodide was added, and the semi was stirred at room temperature for an additional 2 hours. It was then filtered to remove potassium iodide and the Piltrat concentrated in vacuo at about 35 ⁰ C to give a colorless syrupy material, which in a system of ethyl acetate: was subjected to 2 of a counter-current distribution: ethanol + water in the ratio 4: 1. The tractions containing ff-acetyl ^ j ^ O-isopropylidene-TO-methyl-1-deoxylincoeamine (determined by thin-layer chromatography) were combined, evaporated to dryness and the residue was crystallized to give pure H-acetyl ^ i ^ -O-iopropylidene -TO-methyl-l-deoacylincosamine received.

C. 7-0-Ke-OIy l-l-deoxylincosamine. Following the procedure of Example 5, Part C was followed

009820/1916;

BATH

N-acetyl ^ t ^ -O-isopropylidene-T-O-methyl-1-deoxylincosamine converted into 7-0-diethyl-1-deoxylincosamine.

Following the procedure of Example 1, compounds of the formula

XLIV

in which X = R, or Z as defined above) and R has the above meaning.

Example 8; 2-hydroxyethyl-a-thiolincoeaminide.

A. Methyl-N-acetyl-2,3,4,7-tetra-O-thionobenzoyl-α-thiolincosaminide.

Methyl-N-acetyl-ethiolincosaminide (IX) dissolved in dry pyridine is treated with excess thiobenaoyl chloride (180% excess) at ^{0 0} O, and the reaction mixture is left under overnight at room temperature

0 0 9 8 2 0/1916 ⁸ AD ORiGiNAL

Allowed to stand with exclusion of atmospheric moisture. After the most complete removal of volatile ones Fraction at 3O ° C in a high vacuum, the reaction product with Chloroform extracted, the extract is saturated with water, dilute sulfuric acid (l-normal), water aqueous sodium bicarbonate solution and again with water washed and dried over anhydrous sodium sulfate. Removing the chloroform in vacuo gives Methyl-N-aeetyl-2,3,4 ·, 7-tetra-O-thionobenzoyl-a-thiolincosaminide (X), which can be crystallized from acetone or ethyl acetate with the addition of Skellysolve B.

B. 2-Hydroxyethyl-N-acetyl-2-0-benzoyl-3,4-, 7-tri-O-thionobenzoyl-oc-thiolincosaminide.

The tetrathionobenzoate (X) prepared according to Part A, is dissolved in ethanol-free chloroform (to form a 2 to 10 # solution) and with a bromine solution (2 molar equivalents) in chloroform (1-2 # solution, volume / volume) while stirring with a magnetic stirrer Treated at room temperature and exclusion of atmospheric moisture. After 2 hours, volatile components become

009 8 20/1916 BAD OFUQiNAL

as far as possible by evaporation at 30 ⁰ C and 15 mm Mg removed; then further chloroform (approx. 100 ml per 2 g of starting material) is added and removed as described above. Acetone is added to the residue (approx. 25 mm per 2 g of starting material) and triethylaain (2 molar equivalents) and the resulting solution is refluxed on the steam bath for one hour with exclusion of moisture into the cyclic intermediate product (XII).) The cooled solution is then added potassium carbonate (5 molar equivalents), sodium bicarbonate (amount by weight equivalent to potassium carbonate) and water (approx. 10 ml per gram of potassium carbonate), whereupon with ethylene iodohydrin (preferably approx. 6-10 molar equivalents) is added; then the reaction vessel is closed and shaken for 3 hours at room temperature. It is then extracted with chloroform and the chloroform solution is washed with water to remove inorganic salts; then it is dried over anhydrous sodium sulfate. After removal of the chloroform at 30 ⁰ C in vacuum to obtain 2-hydroxy-ethyl-N-acetyl-2-0-benzoyl-3,4,7-tri-O-thionobenzoyl-a-thiolincosaminid (XIY).

BAD ORlGiNAL 009820/1916

The above acylated compound (XIV) is hydrolyzed with hydraain (20 to 50 times the amount by weight, based on the ester) Refluxed for 12 to 36 hours, whereupon excess Hydrazine hydrate is distilled off under reduced pressure. The residue is triturated with acetonitrile and the remaining solid product is filtered off, washed with acetonitrile and the ethanol is recrystallized. Receives one 2-hydroxyethyl-a-thiolincosaminide.

If a 2-alkoxyethyl iodide is used, then in the above process the corresponding 2-alkyloxyethyl compounds obtain.

Following the procedure of Example 1 are also the compounds of formula

XLV

τ—

H ^N

RH

₍ SCH ₂ GH ₂ -OR ₄ OH

009820/1916

in which X »R, or Z as defined above, and R and have the above meaning.

Example 9: Alkyl-α-thiolincosaminide.

XLVI

S-alkyl

If you work according to the procedure of Example 8, but replacing the ethylene;) odhydrins by ethyl iodide, so ethyl-a-thiolincosaminid is obtained, which with the Lincomycin C obtainable according to Example 2 is identical. With other alkyl iodides, the corresponding alkyl ethiolincosaminides are formed obtain.

Following the procedure of Example 1, compounds of the formula

X H

CH.

RH

HO NH HO,

XLVII

S-alkyl

OH

BAD ORiQiNAL

009820/1916

in which X = R ₁ or Z as defined above and R has the above meaning,

Example 10: 2-0- and 7-Q-alkylation of alkyl

(X- and ß-thiolinoosaminides.

XIiVIII

S-alkyl

A. Methyl-1-acetyl-3 »4-O-i-sopropylidene-a-thiolincoaaminide and oxazoline therefrom.

16 g of methyl N-acetyl-a-thiolincosaminide were finely powdered and suspended in 1600 ml of dry acetone while stirring vigorously with a magnetic stirrer. 16 ml of concentrated sulfuric acid were added to the suspension. The suspended solid began to dissolve and in up to 60 minutes the solution was complete. After 3 atündigem standing at room temperature (24-26 ⁰ G) The solution was cooled overnight in a refrigerator at about 5 to 10 ⁰ C.

BATH ORIGINAL

0 0 9 8 2 0/1916

The pale yellow solution was then neutralized by introducing dry ammonium gas with stirring. The precipitated ammonium sulfate was filtered off and washed with acetone. The acetone-Wasohlösungen were added to the colorless Piltrat, whereupon it was ^{evaporated in a drum evaporator at 30 0} C and 15 ml Hg; a mixture of syrup and crystalline pests was obtained. The syrup was dissolved by stirring the mixture with 20 ml of water; the crystalline solid was filtered off and washed with ice-cold water. After drying at 160 ⁰ C and 15 mm Hg 7.12 g solid was obtained of melting point 189-192 ⁰ C. Recrystallization from acetone-Skellysolve B-hexane gave colorless needles of the oxazoline of methyl-N-aeetyl-3,4-O-isopropylidene-a-thiolincosaminide; Melting point 191 to 192.5 ⁰ C, optical rotation [α] ^ χ + 126 ° (ο = 0.8508, ethanol).

Analysis; Calculated for

C: 62.95; H: 7.30; N: 4.41; S: 10.10; 0: 25.20; found:

C: 52.77; Hj 7.34; N: 4.40; Sx 10.12; 0: 25.11.

The aqueous mother liquors were evaporated in vacuo to give a solid consisting of

BATH ORIGINAL 009820/1916

Acetone was recrystallized »It consisted of methyl-N-acetyl-3,4-0-isopropylidene-a-thiolincosaminide with a melting point of 178-180 ⁰ O; optical rotation [<O ² J .s + 189 ° (c = 0.5137 water).

B. Conversion of the oxazoline from methyl-N-acetyl-3,4-isopropylidene-a-thiolincosaminide to methyl-N-acetyl-3,4-0-isopropylidene-a-thiolincosaminide.

Its solution of the above-described oxazoline in 20 ml of hot water was refluxed for 2 hours, whereupon thin-layer chromatography on silica gel showed that the conversion of the starting material into methyl-N-acetyl -3,4-0-iaopropylidene-a-thiolinco8aminide had taken place . Water was then removed under vacuum at 4O ⁰ C, remained after which a colorless crystalline solid, the water hot from a small volume was recrystallized; colorless threads of methyl-N-acetyl-3 »4-O-isopropylidene-a-thiolincosaminide with a melting point of 178-18O ⁰ O were obtained; optical rotation [oc] ² | * + 190 ° (c - 1.223 water).

0. Methylation of methyl-N-acetyl-3,4-0-isopropylidenethiolin-coeaminide.

1.16 g of potassium metal were dissolved in 100 al of tert-butyl alcohol (previously dried over sodium) with stirring on the back

09820/1916 ORIGINAL BATHROOM

river loosened. Bas solvent was used as far as possible removed by distillation at normal pressure and then by distillation in vacuo of 15 mm Hg. The dry solid The residue was added to 100 ml of dry benzene, which was distilled off, leaving a fine powder behind. This was treated again with benzene and again was distilled off in order to completely remove the tert-butyl alcohol.

200 ml of dry benzene were added to the powdery potassium tert-butoxide obtained in this way, and that Mixture was made with a magnetic stirrer at room temperature stirred until an opalescent suspension resulted. These 5 g of methyl N-aoetyl-3,4-0-isopropylidene-a-thiolincosaminide was added and the mixture was allowed to stand overnight stirred at room temperature, after which all solid had dissolved.

The mixture thus obtained was 4-2.4 g (18.6 ml) Methyl iodide was added and it was at for 1 1/2 hours Room temperature stirred. After 1 hour the mixture gave a neutral reaction against damp pH paper. Potassium iodide was made from the

BATH ORIGINAL 009820/1916

The reaction mixture is filtered off and washed with benzene; the washing solution was added to the filtrate. Piltrate and washing solution were then evaporated in vacuo at 35 ⁰ O , whereby a colorless syrup was obtained, which was subjected to Gregenstrom distribution using the ethyl acetate / ethanol / water system in a ratio of 4: 1: 2. After 500 passes, the three components were completely separated, as can be ascertained by I-thin-layer chromatography, the main components being methyl-N-acetyl-3,4-0-isopropylidene-7-0-methyl-a-thiolincosaminide and methylli-acetyl-3,4- 0-isopropylidene-2-0-methyl-a-thiolincosaminide. Methyl-N-acetyl-3,4-0-isopropylidene-2,7-di-O-methyl-ethiolincosaminide represented a smaller proportion.

Removal of solvents from tube No. 250-310 (K = 1.30) gave a glass-like material which crystallized from Xthylaeetat: Skellysolve B-hexanes, methyl-N-acetyl ^^ - O-isopropylidene ^ -O-methyl-octhioline cosaminide in the form of short, colorless Received prisms of melting point 176-177 ⁰ O; optical rotation Ca] ² Jp: + 176 ° (c = 0.6220, chloroform).

Analysis: Calculated for O ₁₅ H ₂₇ NOgS:

C: 51.57; H: 7.79; N: 4.01; S: 9.17; OMe: 8.88; found:

0: 51.82; H: 8.10; N: 4.08; S: 8.94; OMe: 8.49.

BATH ORIGINAL

009820/1916

After removing the solvent from the combined Contents of tube no. 330-384 (K «2.52) by distillation a glass-like product was obtained which slowly crystallized on standing. Recrystallization from ether gave Agglomerates of small, colorless needles made from methyl-N-acetyl-7-Q-methyl-3-4-isopropylidene-a-thiolincosaminide.

Likewise, when the solvent was evaporated from the contents of tube no. 410-450 (K * 5.67) obtained a glass-like colorless product. Recrystallization from ether gave thick needles of methyl-N-acetyl-2,7-di-0-methyl-3,4-0-isopropylidene-a-thiolincosaminide with a melting point of 124.5-126 ⁰ O; optical rotation,

£ ⁵ : + 184 ° (c = 0.8390, chloroform). Analysis: Calculated for C ^ gHggNOgS:

0: 52.88; H: 8.04; N: 3.85; S: 8.82; OCH ₅ : 17.08;

found:

C: 53.02; H: 7.95; N: 4.05? Si 8.73; OCH ₃ : 15.92.

0. Methyl-N-acetyl-2-O-aethyl-a-thiolincosaminide.

A mixture of 2 g of methyl N-acetyl-2-O-methyl-3.4-0-isopropylidene-a-thiolincoeaminid in 50 ml of 0.25 N hydrochloric acid was treated with a Magnetrühr it at room temperature (about 25 ⁰ C ) touched. The solid starting material dissolved in a few

BATH ORIGINAL 009820/1916

Minutes. After 1 3/4 hours, thin-layer chromatography was carried out found that there was no starting material left.

The strongly acidic solution was then stirred using a polystyrene quaternary ammonium anion exchange resin until the colorless, supernatant solution reacted neutrally to pH paper. Filtration, washing the resin with water and removing the water from the piltrate and washing solutions in vacuo gave 1.68 g (95%) of a colorless crystalline residue, which was crystallized from methanol-ether with the formation of long, colorless, matted needles of methyl -N-acetyl-2-O-methyl-a-thiolincosaminid of melting point 237-238 ⁰ C.

Analysis: Calculated for C ^ pHg ^ NOgS:

G: 4-6.56; H; 7.4-9; N: 4.53; S: 10.36; found:

C: 46.72; H: 7.44; N: 4.37; S: 10.34.

D. Methyl-2-0-methyl-α-thiolincosaminide.

The 'methyl-N-acetyl-2-O-methyla-thiolincosaminide obtained in this way was refluxed with 6 ml of hydrazine hydrate for 22 hours. The excess hydrazine hydrate

BAD ORSGiNAL 0 0 9 8 2 0/1 9 T 6

was distilled off in vacuo and the residue became three times recrystallized from ethanol-water, methyl-2-0-methyi-oc-thiolincosaminide received.

E. Methyl 7-0-methyl-α-thiolincosaminide.

In the manner described in Part 0 and D, methyl-N-acetyl-7-O-methyl-3,4-O-isopropylidene-ethioline cosaminide was obtained hydrolyzed and then hydraiinolyzed to obtain methyl 7-0-methyl-α-thiolincosaminide.

F. Methyl ^^ - di-O-methyl-a-thiolinccsaminide.

In the manner described in rope C and D, methyl-N-acetyl-2,7-di-O-methyl-5,4-0-isopropylidenea-thiolincoeminide was obtained hydrolyzed and then hydrazinolyzed to obtain methyl 2,7-di-0-methyl-α-thiolincosaminide.

If the methyl-a-thiolincosaminide is replaced by the ß-compound or other alkyl-a- or ß-thiolincosaminides of the type described above, the corresponding alkyl-2-O-, 7-0- and 2,7-di-O-alkyl-a- and ß-thiolincosaminides obtain.

Each following the procedure of Example 1 will make compounds the formula

009820/1916 ^ßAD

XIIX

in which X = R-, or Z as defined above and E has the above meaning and in which one of the radicals Rg and R "is an alkyl radical with not more than 12 carbon atoms * atoms, while the other is hydrogen or an alkyl radical having not more than 12 carbon atoms means obtained.

Replaces the methyl-lf-acetyl-a-thialincoeaminide by 2-hydroxyethyl-a- and ß-thiolincosaminide, so are the corresponding alkoxyethyl compounds obtained.With replacement from liethyl-N-acetyl-a-thiolincosaminide by 2-trityloxy-. Ethyl-N-aoetyl-a- or ß-thiolincosaminide is obtained the corresponding 2-irityloxyethyl-N-aeetyl-2-0-, 7-0- or 2,7-di-O-alkyl-α- and β-thiolincosaminide compounds. If you remove the trityl group by hydrolysis with 80 iger aqueous acetic acid and the N-acetyl group duroh hydraainolysis,

009820/1916 bad ORIQ'NAL

so the corresponding a-hydroxyethyl ^ -O-, 7-0- and 2,7-di-O-alkyl-α- and ß-thiolincoaaminides were obtained.

Following the procedure τοη Example 1 are also
Compounds of the formula

in which X = R ₁ or Z as defined above, R and R ^ have the above meaning and at least one of
Rg or R ₇ radicals is an alkyl radical having not more than 12 carbon atoms, while the other is hydrogen or an alkyl radical having not more than 12 carbon atoms.

Example 11: Methyl-ö-amino-e ^ -dideoxy-l-thio-L-threo-

α-D-galacto-octopyranoside.

CH,

HO

L I

vOH

_ | / ke OH

009820/1916

BATH ORIGINAL

A. 3 »4-0-IsopΓopyliden-lincomycin.

' NS,

L II

A solution of 9.8 g of lincomycin in 150 ml of acetone is added to a solution of 9.8 g of p-toluenesulfonic acid monohydrate in 100 ml of acetone with thorough stirring and avoiding the addition of moisture. The mixture is stirred at room temperature for 1 hour ₉ whereupon 100 ml of anhydrous ether are added $ then it is stirred again in an Eiefcad for an hour. The mixture is filtered and the solid is ^{dried in vacuo at 50 0} O; Yield 13.35 g (85 * 5 Jt ') of 5 _f 4-0-isopropylidene-lincomycin-p-toluenesulfonate. More I ₉ 15 g (7.4 x Jt) can be obtained from the mother liquor by the addition of 350 ml of anhydrous ether and one hour, cooling the solution. The 14.5 g of product thus obtained are dissolved in 200 ml of ether

009820/1916

BAD ORlGiNAt

suspended and vigorously shaken with 125 μl of 5% potassium bicarbonate solution * The aqueous layer is extracted with two portions of 100 ail ether each. The ether 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 a residue τοη 7.9 g (73.1 fi) 3 »4 ~ -0-isopropylidene-lineomycin, which is dissolved in 25 ml of ethyl acetate, whereupon the solution to a volume τοη about 10 until 15 ml is concentrated. The concentrate is left to stand at room temperature for several hours and then kept in the refrigerator over a compartment. The crystalline material thus obtainable is filtered off and carefully washed with cold ethyl acetate. Is obtained ^ »55 g (42.2 $ 5 3" 4-O-Ieopropyliden-linoomycin from Sohmelspunkt 126-128 ⁰ Cf optical rotation t "1 ² p: 101-102 + °

(c * 1, methylene chloride).

B. 7-Dehydro-3t4 ~ 0 ~ isopropylidene ~ lincomycin.

Su a solution of 6 g (0.0135 mol) of 3,3-O-isopropylidene-lincomyeiii in 75 ml of pyridine were 12 g (excess) Ohromtrioxyi admit it. The temperature of the solution rose about 20 ° ö. After an hour the mixture became one

009820 / 1-9.16-

BATH ORIGINAL

$ 40

Containing 250 al ethyl ether and 250 al ethyl acetate added it was filtered and the ingredient was evaporated, whereby Mn received 3.4 * g of a syrup. This syrup was subjected to a Jegenstroaire division with 500 transitions, the system being water, ethyl acetate, ethanol Oyelohexane (lsisi) was used. From the B @ hro & en Ho. 330-380 (K «2.45) 7-dehydro-3» 4-0-isopropylidene-linoomyoin was isolated »

Analyzes Calculated for ^ «^ H ^ glgOgSs

Os 56.72; Es 8.16g Is 6.30 {Ss 7 _; 21 | foundj Oi 56.37! Hs 7,62} Is 6,51? Ss 6.84.

0. 3,5-O-isopropylidene-epilinoomycin.

OH

I »II

NS,

003820 / $ 111

BATH ORIGINAL

400 ag of sodium borohydride were added to 1.6 g of pure 7-2> ehydro "-3f4-0-isopropylidene linoomyoln la 75 ml of methanol. For 1 * 5 hours the IiiJarüng was evaporated in eiaeK drum _ rerdampfer sur Trooine. Btr Hüokstand was "iiigegebsii" in 25 si water, whereupon drsiaal kit j "25 Kl lethylene chloride was extracted. Kit 15 κΐ water ruokgewasoänen ₉ Heer K% gaaaitÄeiaorid was trooknet per extract and added sur frozen. The huoketaad (1.4 g) was subjected to a power distribution kit of 500 passes, with ethanol (111 ti ti) being used as the solution of water. A single MaxiKUK was observed at K »1.05» which corresponded to the theory. The material located in the racks 240 * 280 was isolated as a syrup.

Analyzes Calculated for ° 21 ³ 38 ^I 2 ^Ö S ^Sl

Os 56.47 | Hs 8.58; Is 6.27} Ss 7.18; found

0s 56.24? Hs 8.54; Is 6.13? Ss 7.01.

Bur eh BunnsohiohteAdhroKatograpMe it was proven that this material consisted of two products, namely 5,4-0 ~ isopropylidane-llnooKyQia «md 3,4-0-isopropylidene -

0 G 9 S 2 8 / 111S ^BAD original

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BATH

Analysis: Calculated for C, «H ,,. N ₀ O ^ S:

Xo 0 * * - ο

0: 50.92; H: 8.55; N: 6.60; S: 7.56; found: G: 50.19; H: 7.91; N: 6.05; S: 6.42.

Fractions 73-104 gave lincomycin at a level of 950 mcg / mg.

B. methyl epi-a-thiolincosaminide.

While carrying out the hydrazinolysis according to Example 1, Part B, Bpilincomycin was converted into methyl-epi-athiolincosaminide converted.

Lincomycin can be replaced by methyl-K-acetyl-a-thiolincosaminide in the above conversions. The above procedure can also be applied to other 6-acylamino- 6 , 8-dideoxy-D-erythro-D-galacto-octopyranose compounds of the formula

LIV

001820/1916

BATH ORIGINAL

can be applied, with the formation of the corresponding epimers:

OH

H ₂ N-HO

.0H

OH

OR-

H Y

which can be alkylated by the above process, under Formation of compounds of the formula:

LVI

In this way, 6-amino-6,8-dideoxy-L · - threo-D-galacto-; 6-amino-2-0-alkyl-6,8-dideoxy-L-threo-D-galacto-} 6-Amino-7-0-alkyl-6,8-dideoxy-L-threo-I> galactound ß-Amino-ajT-di-O-alkyl-oie-dideoxy-L-threo-D-galacto-octopyranoe compounds are produced, which in turn again according to the method of Example 1, Part C

00 9820/1916

BAD ORSG'NAL

and the following, can be treated to form compounds of the formula

LVII

The intermediates of the formula III can be used as buffers or as acid antagonists. they react with isocyanates to form urethanes and can therefore be used to modify polyurethane resins will. The thiocyanic acid addition salts result on condensation with formaldehyde, resinous materials which are found to be pickling inhibitors according to U.S. Patents 2.425.520 and 2.606.155 are suitable. The free bases are also good carriers for toxic acids. For example The fluosilicic acid addition salts are suitable as moth repellants according to U.S. Patents 1,915,331 and 2,075,359; the hexafluoroarsenic acid and hexafluorophosphoric acid

0 0 9 8 2 0/1916

BAD ORIG-'NAL

Addition salts can be used to combat parasites according to U.S. Patents 3,122,536 and 3,122,552.

Various acid addition salts of the intermediates Formula III can be obtained by neutralizing the free base with the corresponding acid at a pH value below about 7.0, preferably at pH 2 to 6. Suitable acids for this are e.g. hydrochloric acid, sulfuric acid, Phosphoric acid, thiocyanic acid, fluokiesiic acid, hexafluoroarsenic acid, heiafluorophoreic acid, acetic acid, succinic acid, citric acid, lactic acid, maleic acid, fumaric acid, Pamoic acid, choic acid, palmitic acid, soleic acid, Capmheric acid, glutaric acid, glycolic acid, phthalic acid, tartaric acid, Lauric acid, stearic acid, salicylic acid, 3-phenylsalicylic acid, 5-phenylsalicylic acid, 3-methylglutaric acid, orthosulfobenzoic acid, cyclohexanesulfamic acid, cyclopentanopropionic acid, 1,2-cyclohexanedicarboxylic acid, 4-cyclohexenecarboxylic acid, Octadecenylsuccinic acid, octenylsuccinic acid, methanesulphonic acid, benzenesulphonic acid, heliantic acid, Heinecke's acid, Dimethyldithiocarbamic acid, cyclohexylsulfamic acid, hexadecylsulfamic acid, octadeoylsulfamic acid, sorbic acid, monochloric

009820/1916 BAD ORIGSNAL

acetic acid, undecylic acid, ^ -'- hydroxyazobenzene - ^ - sulfonic acid, Octyldecylsulfuric acid, picric acid, benzoic acid, cinnamic acid and the same.

The acid addition salts can be used for the same purposes as the free bases; can also they serve to purify the free bases. For example, you can convert a free base into an insoluble salt like Picrat convict, which is subjected to cleaning operations therein, for example solvent extractions, washing, chromatography, fractionated liquid-liquid extraction and Crystallization; the free base can then be regenerated by treatment with alkali or with another salt with metathesis. You can also use the free base in a water-soluble salt, e.g. the hydrochloride or sulfate, and the aqueous solution of the salt with various Extract with water immiscible solvents, before the free base by appropriate treatment of the so extracted acidic solution is recovered.

Typical compounds prepared according to Examples 4, 8 and 9 include the cis and transform of the following compounds of the formulas XXXVII, XXXVIII, XLV and XLVII.

0 0 9 8 2 0/1916 bad original

HH a) OHj b) CHj ο) CHj d) O ₂ H ₅ β) C ₂ H ₅ f) Ji-G ₃ E ₁ G) Γοί i) HO ₄ H ₉ a) HO ₄ H ₉ k) n-0 ₄ H ₉ 1) HO ₄ H ₉ ■) OHj n) OHj O) CHj P) C ₂ H ₅ 0.) O ₂ H ₅

Table I;

R ₁ R ₄ alkyl

H H

CHj H

CHj CHj

H H

O ₂ H ₅ O ₂ H ₅

r) n-C-H ,, H H

CHj OH

O ₂ H ₅

O ₂ H ₅ C ₂ H ₅

H G

H OHj

OH- »CH,

C ₂ H ₅ O ₂ H ₅

HO ₂ H ₅

O ₂ H ₅

0 0 9 8 2 0/1916 BAD QR \ G »NAL

ns

RH R ₁ \ alkyl

a) n-CjH « CHj t) η— G TrRrJ CHj u) H τ) n-0 ₄ H ₉ OH, y) HC ₄ H ₉ OHj

CHj

H H

OHj

Typical ones that can be produced according to Examples 5-6 and 7 Compounds include the cis and trans forms of the. following Compounds according to formulas XL, XLII and XLIV.

Table II:

RH

a) OHj H b) CHj CHj c) CHj C ₂ H ₅ d) C ₂ H ₅ H e) C ₂ H ₅ CHj f) O ₂ H ₅ O ₂ H ₅ G) n-Ojtty H H) n-OjHy OHj i) n-CjH- O ₂ H ₅

0 0 9820/1916

HH R ₁

3) ^η - ^σ 4 ^Η 9 ^Η

k) n-0 ₄ H ₉ CH ₃

Typical compounds according to formulas XLIX and L _f which were obtained by the method τοη Example 10 include the compounds listed above, in which the ZO hydrogen atoms (H ₇ ), the 7-O _{hydrogen atoms (R 6} ) and hydrogen atoms in the form Rg and E _{7 are} replaced by alkyl.

Typical compounds of Formula LVII include also the above compounds with an L-threo configuration.

Example 12: Conversion of lincomycin hydrochloride into

the S-ethyl analog of lincomycin hydrochloride.

8.85 g of lincomycin hydrochloride (0.02 mol) in 20 ml of water were ^{cooled to 0} ° C., whereupon 3.52 g of bromine (0.022 Hol) were added dropwise over a period of 1 minute with stirring. 25 ml of ethanethiol were then added and the mixture was ^{stirred at 25 °} C. for 2 hours. Pas clear, colorless, two-phase system (ethanethiol is related to water

© AD OR1G «NAL 00 9820/1916

insoluble) was cooled in an ice bath * whereupon about Hydrogen chloride gas was introduced for 5 minutes. the The lower aqueous phase turned red. The reaction mixture was then three times with 100 ml Skellyeolve B extracted and an aqueous sodium hydroxide solution was obtained to adjust the aqueous phase to pH 11. The base phase was then extracted well with chloroform. The chloroform extracts were washed with saturated sodium chloride solution, dried and evaporated in vacuo, whereby 6.2 g of a white solid was obtained. From this solid, 4.8 g of 15 g of silica gel were in Methylene chloride adsorbed and the silica gel thus treated was applied to a column 6.2 cm in diameter containing 800 g Abandoned silica gel. The column was then filled with a mixture of methanol and chloroform in a ratio of 1: 7 eluted. After 800 ml forerun, 80 fractions of 25 al each were collected. The fractions 40 to 58 were combined, evaporated to dryness and the residue was recrystallized from acetone. About 0.5 g was obtained 6,8-Dideoxy-6- (trans-1-methyl-4'propyl-L-2-pyrrolidinoarboxamidoJ-D-erythro-D-galacto-aldehydro-octose diethyl dithioacetal of the following formula:

BATH ORIGINAL 009820/1916

> SEt

Analysis: Calculated for

^ 2 ^ 6

O: 52.25; H: 8.77? Ni 5.80; S: 13.29; found: O: 52.16; H: 8.77; N: 5.84; S: 13.30.

Fractions 65 to 75 were also combined, evaporated to dryness and the residue was converted into the hydrochloride transferred and recrystallized from aqueous acetone, about 0.5 g of the S-ethyl analogue of lincomycin hydrochloride, Ethyl-6,8-dideoxy-6- (trans-l-methyl-4-propyl-L-2-pyrrolidincarboxamido) -l-thio-D-erythro-a-

24 ο received D-galacto-octopyranoside hydrochloride; [a] Z : + 147

(c = 1.1206 in H ₂ O). Analysis: Calculated for

0: 49.93; H: 8.16; 01: 7.76; N: 6.13; S: 7.01; found: 0: 49.53; H: 8.23; Gl- * 7 »39; N: 6.04; S: 6.88.

009820/1916

BATH

The product has about twice the bactericidal effectiveness of lincomycin; it is identical to the S-ethyl analogue of Lincomycins (Lincomycin G), which is produced by carrying out the Fermentation according to U.S. Patent 3,086,912 in the presence of Ethionine is obtained.

Example 13: Conversion of Lincoaycin hydrochloride to the

S-isopropyl analogs.

35.4 g of lincomycin hydrochloride (0.08 mol) were dissolved in 80 ml of water, the solution was ^{cooled to 0} ° C. and, with stirring, 14.08 g (0.088 mol) of bromine were added dropwise over the course of 5 minutes. Then, 80 ml of 2-propanethiol and 1400 ml of tetrahydrofuran was added and Ohlorwasserstoffgas was initiated quickly, had risen to the temperature at 43 ⁰ O. At this temperature the two-phase system became homogeneous. The addition of hydrogen chloride gas was then stopped, and the solution was stirred for two hours without external heating or cooling. Tetrahydrofuran was then evaporated off in vacuo and the aqueous residue was worked up as described in Example 12, the 3-isopropyl analog of lincomycin hydrochloride being obtained. This connection showed about that

009820/1916 BAD original

same antibacterial effectiveness as Lincomyeinhydrochlotid \ optical rotation Ca] ² ^: + 137 ° (σ = 0.6798,

Analysis: Calculated for

G: 50.99? H: 8.35; N; 5.95J S: 6.81? 01 »7.53; found: 0: 51.23; Hs 8.39; H: 5.92? S: 7.02? Cl: 7.58.

example

If the procedure of Example 13 is followed, but replacing the ethanethiol with butanethiol, the S-butyl analogue of lincomycin hydrochloride is obtained. This product exhibits about double the antibacterial potency of lincomycin; the optical rotation is [cc] ^.: + 130 ° (c = 0.4070, H ₂ O).

Analysis: Calculated for 0 ₂ l ^H 40 ^li 2 ⁰ 6 ^S · ^flClt

Oi 51.99? Hi 8.52? Nt 5.78? S: 6.61? found: C: 50.64? Ht 8.33} Hs 6.68? Ss 6.41.

Example 15:

If the procedure of Example 13 is followed, but replacing the ethanethiol with cycloheianthiol, in this way the S-cyclohexyl analogue of idncomycin hydrochloride is obtained. This compound shows about twice the antibacterial effectiveness of lincomycin? optical rotation

BAD 009820/1916

[α] * ⁵ : 123 ° (c = 0.9952, H ₂ O).

Analysis: Calculated for C ₂ -H ^ ₂ N ₂ OgS. HCl:

C: 5.54} H: 8.48; N: 5.48; Si 6.27; Cl: 6.94; Found: C: 53.57; H: 8.74; N: 5.37; S: 6.25; Cl: 6.66.

If the alkane thiols used in Examples 12, 13 and 14 are replaced by other alkane thiols, for example by propane, pentane, hexane, heptane, octane, nonane, decane, undecane, dodecanethiol or their isomers, so the corresponding alkyl-6,8-dideoxy-6- (transl-methyl-4-propyl-L-2-pyrrolidine-carboxamido) -l-thio-D-erythro-a-D-galacto-octanopyranoside obtain.

By hydrazinolysis or when applying the above process steps to the free sugar hydrochloride, that no acyl group on the amino nitrogen in the 6-position the corresponding alkyl-6-amino-6,8-dideoxy-1-thio-D-erythro-a-D-galacto-octopyranoside obtain.

Lincomycin C is also obtained by reacting clay lincomycin with Ethanthiöl (ethyl mercaptan) under Formation of a diethyldithioacetal and heating of the the latter in the presence of p-toluenesulfonic acid or Er-

BATH ORIGINAL 009820/1916

hit to the melt. This procedure is described below Example explained:

Example 16:

A. 6,8-Dideoxy-6- (trans-1-methyl-4 ~ propyl-L-2-pyrröli-

dincarboxamidoJ-D-erythro-D-galacto-aldehydo-octose-diethyldithioacetal.

Set

In a 1 liter three neck flask, 150 ml was concentrated Hydrochloric acid and 50 ml of ethanethiol (previously cooled to 0 ° ö) and then 1510 g of lincomycin hydrochloride entered. After stirring for 5 hours at room temperature with a magnet stirrer, the reaction mixture became an equal volume Ice water was diluted and the solution was carefully extracted with Skellysolve B; the extracts were discarded.

0 0 9 8 2 0/1916

BATH

The major portion of the acid was neutralized by careful addition of solid potassium hydroxide (100 g), keeping the temperature of the well stirred Heaktionsgemisehes was maintained by cooling with acetone-Troekeneis 20-30 ⁰ C. Solid potassium chloride was then filtered off and washed well with chloroform. More chloroform was added to the piltrate (approx. 150 ml) and the mixture, stirred with a magnetic stirrer, was adjusted to a pH of 10 by adding aqueous sodium hydroxide (2 normal). The chloroform layer was separated, the aqueous layer was carefully extracted with chloroform, the combined extracts were washed twice with water and then dried over anhydrous sodium sulfate. BSIM removal of the solvent at 30 ⁰ C in vacuo a semisolid residue which upon crystallization from acetone 5 "41 g of 6,8-dideoxy-6- (trans-1-me thyl received-4-propyl-Ii-2-pyrrolidin -carboxamido) -D-erythro-D-gslaeto-aldehydo-octose-diethyldithioacetal in the form of colorless, flattened needles with a melting point of 130-132 °. Concentrating the mother liquor gave

009 8 20/1918

a further 1.50 g with a melting point of 129-131 ° (total yield 6.91 g, 42.4% ). ..; .

Analysis: Calculated for O ^ fi ^ gNgOgSg:

Ox 52.25f Hi 8.77; N: 5.80; S: 13.29 * found: 0: 52.38; H: 8.71; N: 5.93; S: 13.46

B. Oyolosis to Lincomycin C.

(a) One part of diethyldithioacetal according to Part A and one part of p-toluenesulfonic acid monohydrate were refluxed in 25 parts of acetonitrile until the product was found to have substantial antibacterial activity. The reaction mixture was cooled and evaporated to dryness and then on silica gel Chromatographed using a solvent mixture of ethyl acetate, acetone and water in a ratio of 8: 5: 1. Fractions 102 to 131 show bactericidal activity, Fractions 105 to 125 were combined, evaporated to dryness, and crystallized from acetone acidified with hydrochloric acid and recrystallized by dissolving in water and adding acetone, crystalline lincomycin O-hydrochloride having a melting point of 14-9-153 ⁰ O being obtained.

009820/1916 BAD ORlG'NAt

(b) The diethyl dithioacetal, obtained according to Part A, was ^{heated to 260 0} O for about 3 minutes, the odor of ethyl mercaptan being detected. On chromatography as described in part B (a), the product gave lincomycin 0 »

00982 0/1916

Claims (1)

Claims; 1. Process for the preparation of a compound of the formula (H) in which R is an alkylidene group with no more than 20 carbon atoms, a cycloalkylidene group having 3 to not more than 8 carbon atoms, or an aralkylidene group with no more than 12 carbon atoms, R “and R, hydrogen or alkyl groups with no more than 12 Carbon atoms, Y hydrogen, S-alkyl with no more than 12 carbon atoms, -SCHgCHgOH or -SCHgOHgOR ^, where R * is alkyl of not more than 12 carbon atoms is, and R-, an alkyl group with not more than 20 carbons 009820/1916 Substance atoms, an Oycloalkylgruppe with no more% le Represents 8 carbon atoms or an aralkyl group with not more than 12 carbon atoms, characterized in that that one is a compound of the formula (A) C-OH
η in which Z is a protective group removable by hydrogenolysis, with a Wittig ¹ see reagent of the formula R m P (C ₆ H ₅ J ₃ (B) in which R has the above meaning, reacts to form a compound of the formula (C) C-OH R ""
0 0098 20/1916 in which Z and R have the above meaning; the connection of formula C hydrogenated in the presence of a platinum catalyst with saturation of the alkylidene group and formation a compound of the formula (D) RH in which Z and R have the above meaning; with a compound of the formula D a compound of the formula (E) in which R «» R? and Y have the above meaning N-acylated to form a compound of the formula 009820/1916 (F) In which. R, Rp, R ,, Y and Z have the above meaning the compound of formula F hydrogenated in the presence of a palladium catalyst with removal of the group Z and Formation of a compound of the formula in which R, R ₂ and R, as well as Y have the above meaning, and the compound of the formula G is alkylated. 009820/1 91 G 2. The method according to claim 1, characterized in that the alkylation is carried out by reacting the compound of the formula & with an oxo compound of the formula HqRqCO »in which R ₈ RgO» is an alkylidene radical with not more than 20 carbon atoms, a cycloalkylidene radical with 3 to not more than 8 carbon atoms or an aralkylidene radical having not more than 12 carbon atoms, whereupon hydrogenation is carried out in the presence of a catalyst which is suitable for saturation of olefinic double bonds. 3. The method according to claim 1, characterized in that one compound of formula A with a Wittig'sohen Reagent of the formula B converts, with the formation of a compound of the formula C, the compound C in the presence of a Platinum catalyst hydrogenated with saturation of the alkylidene group and formation of a compound D, with the compound J) a compound of the formula E N-acylated to form a compound of the formula F and the compound F hydrogenated in the presence of a palladium catalyst with removal of group Z and formation of a compound of formula G. 009820/1916 H-. Process according to Claim 1, characterized in that a compound of the formula A is reacted with a Wittig's reagent of the formula B to form a compound of the formula 0, the compound 0 is hydrogenated in the presence of a platinum catalyst with saturation of the alkylidene group and formation of a compound of the formula S, and with the compound D a compound of the formula S is N-acylated to form a compound of the formula f. 5. The method according to claim 1, characterized in that a compound of formula C is in the presence a platinum catalyst hydrogenates to form a Compound of formula D that a compound of formula E is N-acylated with a compound of formula D, to form a compound of the formula f that the compound f is hydrogenated in the presence of a palladium catalyst with removal of the group Z and formation of a compound of the formula G and that the compound G is alkylated forming a compound of the formula H.. 6. The method according to Anspruoh 5, characterized in that the alkylation is carried out with reaction the compound of formula G with an oxo compound 009820/1916 of the formula RgRgCO, in which R ₈ RgC * is an alkylidene group with not more than 20 carbon atoms, a cycloalkylidene group old 3 here not more than 8 carbon atoms or an aralkylidene group with not more than 12 carbon atoms, and then in the presence of one capable of saturating olefinic double bonds Hydrogenated catalyst. 7. The method according to claim 1, characterized in that a compound of formula C is hydrogenated in the presence of a platinum catalyst with formation a compound of the formula D, 'a compound of the formula E with the compound D H-acylated to form a Compound of Formula F, and the compound of Formula F in the presence of a palladium catalyst with removal of the group Z and formation of a compound of the formula G hydrogenated. 8. The method according to claim 1, characterized in that a compound of formula C in the presence of a platinum catalyst is hydrogenated to form a Compound of Formula D and a compound of Formula B with a compound D H-acylated to form a compound of Formula F. 009 ß? Q / 1 9 1 6 9. The method according to claim 1, characterized in that a compound of the formula A is reacted with a Wittig ¹ see reagent of the formula B to form a compound of the formula C, that a compound of the formula E with a compound of the formula Q N- acylated to form a compound of the formula CH, R ₂ O - HH (D that the compound of formula I is hydrogenated with saturation of the olefinic double bond and removal of the Group Z and to form a compound of the formula G, and that the compound of the formula G is alkylated under Formation of a compound of the formula H. 10. The method according to claim 1, characterized in that a compound of the formula A is reacted with a Wittig ¹ see reagent of the formula B to form a compound of the formula C, that a compound of the formula E is N-acylated with a compound of the formula 0 00 9 820/1916 to form a compound of the formula I ₁ hydrogenating the compound of formula I, below saturation of the olefinic double bond ", and removal of the group Z and Formation of a compound of formula G. 11. The method according to claim 1, characterized in that that a compound of formula E with a compound of formula G N-acylated to form a Compound of formula I, and that the compound of formula I is hydrogenated with saturation of the olefinic double bond and removing group Z and forming one Compound of the formula G. 12. Process for the preparation of a compound of the formula 00 9 820/1916 (AO) in which R is hydrogen or an alkyl radical having not more than 12 carbon atoms, Ac is an alkanoyl or
Aralkanoyl group with not more than 12 carbon atoms or an acyl radical of the formula (AP) in which R is an alkylidene group with not more than 20 carbon atoms, a cycloalkylidene group with 3 to not more than 8 carbon atoms or an aralkylidene group with not more than 12 carbon atoms and R-, hydrogen, an alkyl radical with not more than 20 carbon atoms, is an cycloalkyl group with 3 to not more than 8 carbon atoms or an aralkyl group with not more than 12 carbon atoms, and Y is hydrogen, S-alkyl with not more than 12 carbon atoms, -S-OHgOHgOH or -S-OHgCHgOalkyl 009820/1916 with an alkyl radical of not more than 12 carbon atoms, characterized in that one represents a compound the formula GH « (AK) in which R ,, Ac and T have the above meaning, reacts with acetone to split a compound of formula (Ai) this is oxidized with chromic acid to form a compound of. formula (AT THE) 009820/1916 BAD ORiGlNAt 1HO the compound AM into a compound of the formula AcNH (AT) converted by reduction with a borohydride, and the isopropylidene group by acid hydrolysis and the Acyl group removed by hydrazinolysis. 13. Process for the preparation of a compound of the formula 0 SR 10 (BA) in which R _{10 is} an alkyl radical with not more than 12 carbon atoms, -CH ₂ CH ₂ OH or -CHgCHgOK ^, where R _{4 is} an alkyl radical with not more than 12 carbon atoms, characterized in that a compound of the formula - Cv. * 009820/1916 4H4 NS, (AW) in which Ac ¹ denotes an alkanoyl or aralkanoyl group with not more than 12 carbon atoms, desulfurized with mercury (II) chloride and the product thus obtained acylated to form a compound of the formula (AX) OAc in which Ac ^{1 has} the above meaning and Ac represents an alkanoyl or aralkanoyl group having not more than 12 carbon atoms, that the latter compound is brominated with Hbr to form a compound of the formula 00982 0/1916 BATH OH, AcO · ■ Ac ¹ NH ■ AcO (AY) kOAc Br OAo in which Ac and Ac ^{1 have} the above meanings, the latter compound with thiourea and a compound of the formula R, Q-halogen is thioalkylated to form a compound of the formula (AZ) ÖAc and that the groups Ac are hydrolyzed and the groups Ac ¹ hydrazinolizes to form the compound of Formula BA. 14. Process for the preparation of a compound of the formula OH, AcNH 4 ^) CSO) ° _N / 00S (^ S-R 10 ) 000 (BF) ^BATH 009820/1916 in which R-. q an alkyl group of not more than 12 carbon atoms derived from the alkyl ^G _n ^H 2n ^H ₂₂ or -CH ₂ CH ₂ OR ₄ , where R _{4 is} an alkyl group with not more than 12 carbon atoms, characterized in that one is a compound of the formula CH, HO
AcNH HO OH (BC) ^SC n ^H 2n ^H OH in which Ac is an alkanoyl or aralkanoyl group having not more than 12 carbon atoms and η is an integer from 1 to 2, acylated with a thioobenzoyl halide to form the tetrathionobenzoate, the tetrathionobenzoate brominates to form a compound of the formula CH, (J) CSO -—- AcIiH - ■ Ocso- Br (BD) 0 09820/1916 BATH ORIG> HAL the resulting compound is heated in acetone in the absence of water to form a compound of formula CH, OCSO AcNH -I- (J) CSO _t -0 ^(BE) N ^Br " 0-C - and the resulting compound is alkylated with a compound of the formula R, _o -halo in the presence of a weak base to form the compound of the formula BF. 15. The method according to claim 14, characterized in that subjecting the compound of the formula BF to hydrazinolysis to form a compound the formula CH, HO -J- HO 0 (BS) \ 0H ^X '' ^SR io OH 009820/1916 1620007 in which R _{10 is} an alkyl group with no more than 12 carbon atoms, which differs from the alkyl group CH "J in the starting material, -ÖHgCHgOH or -CHgCHgO ^, where R ^ is an alkyl group with no more than 12 carbon atoms" 16. The method according to claim 1, characterized in that a compound of formula G is alkylated, forming a compound of the formula H » 17 * The method according to claim 16, characterized in that that the alkylation is carried out with reaction the compound of formula & with an oxo compound the formula RgRnCG, in which RqRqC = an alkylidene radical with no more than 12 carbon atoms, a Cyclö alkylidene radical with 3 to not more than 8 carbon atoms or an aralkylidene radical with not more than 12 Carbon atoms, and the product in the presence of one hydrogenated catalyst capable of saturation of olefinic double bonds. for their tJpjöhn Company Ui 'x Lawyer 009820/1916 BAD