DE1620620A1 - Process for the preparation of lincomycin derivatives - Google Patents

Description

1620520

Dr. Walter Beil Alfred Kceppener 28. ^ n.

Dr. HnBS Joachim Wolff Dr. Hans Chr. Beil +

Rechismnvälte

Frankfurt a, M.-Höchst

Adelomtraßc 58 -TeL 31264 »

Our MT. 12th

THE UPJOHN GOi / ΐΡΑΝΪ Kalamazoo (Michigan, VStA)

Process for the preparation of lincomycin derivatives

The present invention relates to a process for the production of 7'-οη1θΓ-7-Λβ ^: οΊΧ7ΐίηοοιη3Γθίη_μηα.- 7-Ohlor «7-deoxy-epilincoinycin and their analogs and isomers.

The compounds obtainable according to the invention be .'-. itzen the following formula

69817/180 8

Oil

AcNH

in which E is an alkyl radical with less than 20, preferably less than 8 carbon atoms, one CyIcLoalkylrest with 3 to at most 8 carbon atoms or an aralkyl radical with at most 12 carbon atoms, preferably at most 8 carbon atoms, while Ac is the acyl radical of a 4 ~ substituted-L-2 ~ PyrrolidincarTDonic acid of the formulas

(A)

OH

(B)

OH

"means in which E _{1 and E 2 are} alkylidene radicals of at most 20 carbon atoms (including methylene), preferably at most 8 carbon atoms, cycloalkylidene radicals with 3" to at most 8 carbon atoms or aralkylidene radicals

radicals having a maximum of 12 carbon atoms, preferably a maximum of 8 carbon atoms, represent, while R ₂ .

Is hydrogen or ESU.

Examples of alkyl radicals with a maximum of 20 carbon atoms (R, IiR ₁ and HR ₂ ) are methyl, ethyl, propyl, butyl, £ βι tyl, hexyl, heptyl, octyl, ifonyl, decyl, T-indecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadeoyl, octadecyl, ITonadecyl and bicosyl radicals and their isomeric forms. Examples of cycloalkyl radicals are Oyclopropyl-,. Oyclobutyl-, cyclo-. Pentyl- _t Cyclohexyl-, öycloheptyl-, Oyclooctyl-, 2-Methyloyclopentyl-, 2,3-Dimethyleyclotiutyl-, 2-Methylcyclotutyl- and 3 * -öyclopentylpropylrest · Examples of aralkyl radicals are the Eenzyl-, Phenäthylpro ·, 3 -yl and 1-naphthylmethyl radical. Suitable alkylidene, cycloallidene and aralkylidene groups (R, una Rg) are, for example, the methylene, ethylidene, fiDpylidene, eutylidene, lentylidene, hexylidene, iiept; Octylidene, lionj'li ^ en-, Decyliden-, UndeqjÜ den-, Do.iecylien-, Tr i decyl ir sn, tetradecylidene, pentadecylidene, hexadecylidene, heptadecylidene, octadecylidene, itonadecylidene, eicosylidene groups such. ö ~ dex ^ en isomeric forms öycloprojyliden, Gyclobutyli-l ^ n, J ^

Ir op / I ät hy 1 id en, j. - Jy e 1 open tv 1 pr opyl i 5 ·; -η, Zen. cyl i Ί e r. , 2-i-henyläthyliren, 3-l ^: hen - irror. ^v Ii - "- n and 1-Ii3p :: thyl :: j? tivie-: i.

0 ■ ü S fc t 7/1 δ b η

BAD OHJGiNAL

The above compounds of formula I and analogs are obtained by adding the 7-H ₁ ? dr oxy group of a compound of the formula

Ac

II

replaced by chlorine »The replacement is expedient by the starting compound of the formula II is mixed with thionyl chloride and heated * The substituents Ac and R can be any radicals that do not react with thionyl chloride, "but have in particular the meaning given above.

For example, if a compound of the formula is used

(Ac in formula II corresponds to the acid of formula A) so he " if you keep a new compound of the formula I A. This is the case Compound or the starting material (formula IIA) with a catalyst for the hydrogenation of olefinic double bonds hydrogenated, a compound of the formula IE ale is obtained 3-einic.ch of eis and tr & ns epimers according to the following formulas:

Q09817 / 18bb bathroom

162062g

OH,

NH-

, 01

CH,

NH-

, 01

III

and

which optionally by countercurrent distribution or on can be separated by chromatography.

If R ₃ in the formulas B, IB and HB is hydrogen, this hydrogen can be replaced by alkylation or the like. Aldehyde or ketone) and the resulting adduct is hydrogenated in the presence of a suitable catalyst. For this purpose, platinum or palladium can be used as catalysts. Suitable oxo compounds have the formula R ^ R ₁ -CO, in which R ₂ -Rt-O « Examples of such oxo compounds that have the meaning of Ep are: formaldehyde, acetaldehyde, propionaldehyde, butyraldehyde, acetone, isobutyl methyl ketone, benzaldehyde,

-5

0Ö9817 / 1888

BAD OHiGJNAt-

Phenylacetaldehyde, hy dr ο ζ im t al de hy d, acetophenone, propiophenone, butyrophenone, 3-methyl-4-phenyl-2-butanone, 2-methyl-5-phenyl-3-pentanone, rJ-cyclopentane propionaldehyde, cyclohexanacetaldehyde , Cycloheptanecarboxaldehyde, 2.2 _T dimethylcyclopropylacetaldehyde, 2,2-dimethylcyclopropylmethyl ketone, cyclopentyl methyl ketone, cyclobutyl methyl ketone, glyclobutanone, cyclohexanone, ^ -Methylcyclohexanone and the like

The starting materials of the formula II are obtained by making a compound of the formula

OH

OH

in which R has the above meaning with a 4-substituted-L-2-parrolidinecarboxylic acid of formula A or E acylated. The acylation can be carried out by processes common to acylation of amino sugars are known per se. The starting acid of the formula A can be prepared by adding a 4 ~ oxo compound the formula

- OE

08 9117/1888

in the Z a protective, cleavable by hydrogenolysis Hydrocarbyloxyearbonyl group, e.g. Trityl, triphenylmethyl, Diphenyl (p-methoxyphenyl) methyl, bis (p-methoxyphenyl) phenylmethyl, Eaa is zyl or p-nitrobenzyl, with a vifitt ig 'see reagent, e.g. an alkylidene triphenylphosphorane (see Wittig et al., 1-er., 8 ?, 1348, 1954; Trippett, Quarterly Reviews, XVII, lTo.4, p. 406, 1963). Examples for hydrocarbyloxycarbonyl groups Z are tert-eutoxyoarbonyl, Benzyloxycarbonyl groups of the formula

Il

CH ₂ OG-

in which X is hydrogen, a nitro or methoxy group, chlorine or B: -om represents, e.g. the carbobenzoxy radical, p-nitrocarbobenzoxy radical, p — erom or p-chlorocarbobenzoxy radical, as well as phenyloxycarbonyl groups of the formula

in the X-, hydrogen, an allyl- o ^ '- r alkylreer mi; "not more than 4 carbon atoms ledeauet, like zE, Phenyloyvca p-Tolylox ^'carton;.!, pj * thylrhenj'lo>"'car' :: onyl oiei ¹ p-AlIylphi ::, oxycarton;. "l and ^ rrjlsicrien.

" ⁷ ~ 0 0 9 8 I 77 1 S fb

BAD ORlGiNAL

When carrying out this process, the 4-oxo-L-2-pyrrolidinecarboxylic acid (formula O) is added to the freshly prepared Wittig ¹ Gehen reagent. The Vittig ¹ see reagent used here can be represented by the following formula:

R ₁ - P (0 ₆ H ₅ ) ₃

in the R, has the above meaning. These Wittig reagents are obtained by reacting an alkyl, cycloalkyl or aralkyltriphenylphosphonium halide with a base such as sodium amide or sodium or potassium hydride or the sodium or potassium metalate of dimethyl sulfoxide or the like (For the preparation of phosphoranes see Trippett, Quart., Rev. XVII, Ιίο.4, ρ, 406, 196 ^) The reaction is generally carried out in an organic solvent such as benzene, toluene, ether, dimethyl sulfoxide, tetrahydrofuran or the like, at temperatures between 10 ⁰ C and the return

of the reaction mixture. The product thus obtained, a 4 ~ .Alkyli ^ _t en_ 4-OyclQaXkyliöen- or 4-aralkylidene-L- - L-proline, dae following formula:

)H

009817/1838

is isolated from the reaction mixture in a conventional manner, generally by extraction from aqueous solutions of the reaction mixture. The crude product can be used in a conventional manner be purified, for example by recrystallization, chromatography with the formation and purification of easily prepared derivatives such as the amine salts, e.g. the dicyclohexylamine salts, and the like, whereupon the amino acids are released from this compound »By hydrogenating an acid of the formula D in the presence of a catalyst, e.g. platinum, which catalyst: promotes saturation of a double bond, but does not cause hydrogenolysis, a compound of the following form

OH

For example, one can use platinum on a carrier such as carbon or anion exchange resin such as Dowex-III, a crosslinked polystyrene-trimethylbenzylammonium resin in the hydroxyl form. If desired, the starting compounds of formula V with an acid of the formula 0, D or E may be acylated under ¹ Eildung of compounds 110, HD and HE. The compound HO can then be converted into the compound HD by treatment with an i / ittig ¹ see reagent, and HD hydrogenated to form compound HE. The hydrogenation

0 OS 817/1888

BATH

Both the acid D and the acylate HD lead to a mixture of cis and trans epiiners which, if desired, can be separated by countercurrent distribution or chromatography. The starting acids of the formula B, in which R * represents hydrogen, are obtained when an acid of formula D or Ja is subjected to hydrogenolysis in the presence of a palladium catalystE zE palladium on carbon. In the same way, compounds of the formula HD and HE are converted into compounds HB, in which R * denotes hydrogen Above-mentioned processes in compounds of the formulas B and HB, in which R ^ »HR ₂ , are converted. The starting acids of the formula A are obtained by treating an oil of the formula D with bromine hydrogen in acetic acid with removal of the group Z and replacement of the hydrogen in the iT with a group IiRg, in the manner described above, compounds of the formula HD and HE are converted into compounds of the formulas IIA and IIB using the same procedure

Some of the starting compounds of the formula II is obtained biosynthetically. Lincomycin (-ethyl-6,8-di de oxy-6- (trans-1-methyl-4-propyl-L-2-pyrrolidine arboxy amido) l-thio-D-erythro-Ä — D-galacto-octopyranoside) receives raan as metabolism product of a licomycin-producing actinomyceter

- 10 -

009817/1638 BADc

according to the method of the U * S, patent 3 »036,912 · the following formula t

owns

OH

H0 <

HH

in d ^fi r R and R. ₇ Methyl, RiH Pic ^ yi represents Lincomycin 1, (iiethyl-6,8-dide oxj ^r -6- (trans-1-ynethy 1-4-ath j lL-2-pyrrolidincartoxyamido) -l ~ thio-D-er5 ^r thro'-0L-D-galacto-octopyraiiosid) (formula VI, in which R and R, methyl and Η _χ Η denote ethyl) is also a metabolic product of the same microorganism, when cultivated according to the ancestor according to UB Pattnt 3.036.912. Lincoaycln 0 (ethyl-6, ^Ä -äideo3q7-6- (trans-

erythrp- -ii ^ galacto ^ octopyranoside) (formula VI with H «lth: ^r l, R ^ H * propyl, E, -» methyl) is obtained if the method of the US Pat of added ivbhionine carries out lincomycin D (methyl-o, 8-dideoxy "-6- (trans - ^ - propyl-L ^ -pyrrolidincerboxamido) -1-thip-D-eryiihro-ol-D-galacto-octopyrano oil) (formula VI with R! ■! Ethyl, R, H * Pxnryl and R _x = * water to Tf) vdrd crhaltien, -.c-ru

- 11 -

009817/1888

the fermentation according to U.Ö. Patent 3,086,912 in the presence of OL-MTL, methyl-6-amino-6,8-dideoxy-D-er; ythro-lthio-Gl-D-galacto-octopyranoside, one through Hydrazeinolysis of lincomycin available compound. Methyl- 6,8- * dideoxy-6- (trans-4-ethyl-L-2-pyrrolidincarboxamido) -1-thio-D-erythro-kD-galacto-octopyranoside (formula VI with R m methyl, R ₁ H - Ethyl and R ^ »hydrogen) are also formed when the fermentation according to US patent 3.Ü86.912 O.-MTL is added. Lincomycin K (ethyl-6,8-dideox ^ '- 6- (tr.uns-4-piOpyl-L-2-pyrrolidincarboxamido) 1-thio-D-erythro-ol-D-galacto-octopyr- anoside) (formula VI with E ■ ethyl, R, H »irropyl and Rz * - Ϋ / hydrogen), if the fermentation according to US patent 3,086 * 912 in the presence of (K-iiTL, ethyl-6-amino-6,8 -dideoxy-l-thio-D-eryth.ro «OL-D-galaio-octopyranoside, a compound obtainable by hydrazinolysis of lincomycin G. Furthermore, ethyl-e ^ B-dideo ^ -e-Ctranß - ^ - ethyl-LS-pyrrolidincarboxamid) -1-thio-D-erythroft — D-galacto-octopyranoside) (formula VI with R - ethyl, R ₁ H - ethyl and R, - hydrogen), if the fermentation according to US Pat 3.086.912Ov-BTL is added. The above-described N-desmethyl compounds, which are obtained on addition of Oi-MTL and Ol-ST ij to the Fe na entation, are examples of compounds of the formula HB in which R _{2 is} hydrogen. By replacing the

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0 0 9817/1888

IT — hydrogen atoms in compounds of the formula IiB, compounds with IU m HEg »ZeE, methyl-6,8-dideoxy-6- (transl-ethyl-4-propyl-L-2-pyrrolidine carboxaraido) are obtained by the method described above ~ l-thio - D-erythro-α -D-galacto-octopyranoside, ethyl-6,8-dideoxy-6- (trans-lmethyl - ^ - ethyl-L ^ -pyrrolidincarboxamido ^ l-thio-D-erythro- Ow-B-galacto-octopyranoside, iithyl-6,8-dideoxy-6- (trans ~ ethyl ^ -äfchyl-L-2-pyrrolidinecarboxamido) -l-thio-D-erythi-o-α-D-galaoto-octopyranoside and methyl-6,8-dideoxy-6- (trans-ethyl- ^ - ethyl-L-2-pyrrolidinecarboxamido) -l-thio-I) -erythro-O-D-galacto-octopyranoside.

Lincomycin or starting compounds of Eormel II with D-erythro configuration can be combined with L-threo configuration are converted by oxidation of the 7-hydroxyl group to a 7-oxo group and reduction of the O: ro group to the hydroxyl group. One such procedure is reproduced in the following diagram:

00 9 8 17/1888

BATH ORIGINAL

VII acetone

AcNH

AcIIh

OH

VIII

IX

For example, lincomycin suffered from treatment with acetone in the presence of p-Toluenesulfonc & ure 3,4TG-Isopropylirlen ~ lincomycin, which by oxidation with uhromoXyd in 7 — Dehydro— 3, ^ - O-isojjropylidenlincomycin (iJethyl-e ^ -dideo ^ y ^, 4-0-isopropylidene-6- (trans-l-methyl-4-propyl-L-2-pyrrolidincarboxamido) -l-thio-D- glycero-OL-.D-galacto-octanopyranoc-7-ulosid) is convicted, v; elches in turn by EeL.cjadl.xng with eorhydride in 7 - ^ pilincoiaycin (Meth,. l- €, 8-dideoxy-6- (trans-l-methyl - ^ - propyl-L-P.-pyrrolidincartox " amido) -l-

008817/188

thio-L-threo-CL-D-galacto-octopyranoBid) is converted · All from ganjt; Compounds of the formula II with D-erythroioniiguration can also be converted into these, lastly, into Veitindungen with L-threo configuration.

Since the tiosynt ^ etiech produced Lincomycins just as the amino sugars made from them are either methyl or ethyl thioglycoses, "occasionally exists the desire to convert rie into higher or lower glycosides, just like this occasionally the pail ict "with connections der Foi'ieln I, II or V «The conversion can take place, internally the connection to be converted with a Mercaptan of the formula HgBH converts into; v Icher E ^ an alkyl radical having not more than 20 carbon atoms differing from H by one carbon atom and the resulting Dimercaptal represents cyclized · For example one obtains from Connections of Fora I and II through ümi-etzong with a Mercaptan of the formula BgSH the dithioacetals of the formula

AcHH

AcIiH

or

- 15 -

009817/188 «

XIE

SAD

162062Q

in which X is a hydroxyl group or chlorine which upon treatment with an acid or upon heating, in the presence or absence of acid, are cyclized again to form compounds of the formula

AcIQi

XII

OH

The process can be applied directly to any external connection of formulas II, i.e. IiA, HB, HC, HD and HE are used become «The resulting products can be one of a hydrazine · subject lyee, with compounds of the following formula

OH

XIII

obtained, which can be acylated by the method described above with acids of the formulas A, B, G, D and E,

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009817/1888

^BATH

to form compounds of Formula XII in which -X is a hydroxyl group «The process can also be applied to starting materials of the formula V. For example becomes GL-MTL when treated with ethyl mercaptan and Gyklization ino .'- M'L transferred. ■

Another method of making connections of the formulas XII or XIII consists in that Dian brominates the starting material (formulas I, II or V) and then the product reacts with a mercaptan according to the following reaction scheme:

XVa

009817/188

BATH ORIGINAL

The Aasgangsverbindung XIV is dissolved in water in the form of a salt löolichen ^ example of nydroChlorids solved, '.vorauf under liühlen, preferably added at about -10 ° v "^ u to ⁰ C, bromine; IRDO ÜLS is enough to the aqueous solution Cools to about 0 G and adds Erom drop by drop. The stoichiometric amount of bromine is 1 i.Iol per ml. It is advisable to work with a small excess of bromine of approx. 25 yes «The bromine initially replaces the RS group and the resulting intermediate product hydrolyzes to the sugar, in which the pyranose form XVa is in equilibrium with the aldose form.XVb. In the presence of acid, for example hydrochloric acid or another strong, non-oxidizing acid such as p-toluenesulfonic acid or sulfonic acid anion exchange resins, the mercaptan RgSH reacts with the sugar XV to form the Ihioglycoside XVI. At the same time, some diacetal can be formed which, after separation has taken place as described above, can be cyclized to form the desired Th.io glycoside XVI.

The mechanism by which thionyl chloride causes chlorine to replace the 7-hydroxyl group is not fully understood. However, it is assumed that the configuration will be changed. If this is the

- 18 -

000817/1888

If this is the case, a 7-l! YcU'oxy compound results in the B-erythro configuration a 7 ~ jhlor ~ connection of the L-threo-, Configuration. For example, 7-ghlor-7-deoxyloncomycin, made from lincomycin with D-erythro-IConriguration, the L-threo-configuration solution was initially postulated that the procedure runs through several intermediate stages, according to following scheme:

SOCl,

25ο ^σ σ.

Cl

AeNH

SOOi,

KaOH

XVtIb and XVIII

wherein Ac and R have the above meanings. The intermediate product XVIIa is the 3,4-cyclic sulfite of materials II and has the following formula

009817/1888

BATH

AcNH

162062Q

XVIIa

OH

in which X is a hydroxyl group ". The intermediate XVIIb is the corresponding 3,4-cyclic sulfite of the 7-chloro-Vt bond (Formula I), which corresponds to the formula XVIIa with X «chlorine · The intermediate product XVIII can be a bis-sulfite of the 3,4-cyclic sulfite of the 7 - '^ hIorver "bond, the can be represented by the following formula:

AcKH

C-S

01

o-s-

XVIII

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009817/1888

Original

If necessary, all three intermediates XV ", XVIIa and XVIII can be isolated in the course of the reaction. To replace the 7-hydroxyl group with chlorine, however, it is only necessary to use the starting compound of the formula II or V, conveniently in the form of an acid addition salt, e.g. of the hydrochloride, to be mixed with thionyl chloride, advantageously in the presence of an inert solvent, with gentle heating, preferably at reflux temperature, until the desired substitution by chlorine in the 7-position has taken place.Bas is advantageous, the reaction in an inert atmosphere, for example under nitrogen Carbon tetrachloride can be used as a solvent in addition to other inert solvents such as chloroform, methylene chloride, ethylene chloride, ether, benzene and the like it was as long as the aim he sufficiently clear solution is required, the mixture is stirred at room tempera door and then the temperature increases to 50 to 100 ⁰ C, for example at reflux temperature (77 ° C in carbon tetrachloride). When the reaction has ended, usually after refluxing for 1 to 5 hours, the reaction mixture is cooled, advantageously under nitrogen. The material which precipitates out on cooling is isolated and

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009817/1888

BATH

dried. The solvent is removed in vacuo at flask temperatures preferably less than about 35 °, and the product which precipitates out is collected and dried and then treated with ethanol in order to obtain a liquid product e into the desired end product. The material obtained in this way can be extracted by solvent and / or recrystallization v / grounding further purified; it can be used as a free Ease or in the form of an acid addition salt isolate.

The loyalty ratios of the reactants can fluctuate within wide limits · The toichiometry however, requires at least 3 LIoI thionyl chloride per mole of starting compound. Larger quantities can be used however, it is usually neither necessary nor desirable to use more than about a 10-fold overshoe. Zv; angular 2 to 3 times the amount is used as excess. the The amount of solvent is not critical and depends on the the respective needs of the practice. Usually about 15 to 30 parts by volume of solvent per part of the solid will suffice Output compound. However, the ratio of solvent to thionyl chloride is important because of the solubility of the Product in thionyl chloride. If the volume ratio of solvent to thionyl chloride is high, the desired one falls Product when cooling the reaction mixture from and the

22 -

009817/1888

^ß AD ORIGINAL

This simplifies processing. For example, falls when using carbon tetrachloride for a mixture of ingredients the ke 3k ti ons mix directly as it cools down, if that Volume ratio of carbon tetrachloride to thionyl chloride is kept at about 10: 1

The compounds of the iormeln IA ₁ IE, ILA, HB and V exist in both protonieiter and non-protonated form, depending on the PII tfert the environment. The protonated form is called the acid addition salt, the non-protonated form is called the free base. The free bases can be converted into stable acid addition salts by reaction with appropriate acids at pi -> / erten below about 7 ° C, and form with advantage at pH about 2-6. Acids suitable for this purpose are, for example, hydrochloric acid, sclvefelic acid, phosphoric acid, thiocyanic acid, fluosilicic acid, hexafluoro-arsenic acid, hexafluorophosphoric acid, es ^ i ^ æare, succinic acid, citric acid, lactic acid, maleic acid, fumaric acid, pamic acid, gholic acid, ialmitic acid.

Oamphoric acid, glutaric acid, glycolic acid, phthalic acid, "tartaric acid, Lauric acid, stearic acid, salicylic acid, 3-phenylsalicylic acid, 5-phenylsalicylic acid, 3 - !. iethc / glutaric acid, orthosulf topzoic acid, cyclohexanesulfamic acid, glyclopentahpropionic acid, !, S-cyclohexanedicarboxylic acid, ^ -cyclohexanecarboxylic acid, Ootadeceiij'lsuccinic acid, octenylbarnstinic acid, iiethansulfon-

- 23 -

Q0SS17 / 1888

SAD

acid, eenzenesulphonic acid, lielianthic acid, Reinecke ¹ s acid, dimethyldi-thiocarbamic acid, cyclohexylsulphamic acid, hexadecylsulphamic acid, octadecylsulphamic acid, sorbic acid, lonochloroacetic acid, uhdec, sulfamic acid, zensimonic acid, 4 -'-hydroxyadecylsulfonic acid and the same.

The acid addition salts can be used for the same purposes as the free acids; they can also be used for cleaning the free bases. For example, a free Ease can be converted into a water-insoluble salt, e.g. the picrate, since Godann is purified, e.g. by solvent extraction and washing, chromatography, fractionated liquid-liquid extraction or iciness whereupon the free base is regenerated by treatment with alkali; another salt can also be obtained under metathesis ν earth. Furthermore, the free base can be converted into a water-soluble salt, e.g. Conversion of hydrochloride or sulphate, and extract the aqueous solution of the salt with various solvents which can be used with "aseri", before the free base is regenerated by treating the extracted solution. The free base of the formulas IA, IE, HA, UE and ¹ T can be used as buffers or anti-acids. The compounds of the formulas I, II and V react with isocyanates to form urethanes and can therefore

- 24 -

009817/1888

162062Q IS

can be used to modify polyurethane resins. The long-chain compounds, ie those in which HHp is an alkyl radical with more than carbon atoms, have surface-active properties and can be used as surfactants and emulsifiers. The addition salary with thiocyanoic acid results in the condensation with üj'omalichyd resin-like materials which are suitable as ice inhibitors according to the FoSe patents 2 _β 425 · 320 and 2,606,155. The free bases are also good for toxic acids. For example, the fluosilicic acid addition salts are suitable as moth repellants according to UB patents 1.915 "334 and 2.075 o359" while the hexafluoroarsenic acid and hexafluorophosphoric acid addition salts are parasiticides according to US patents 3,122 * 536 and 3,122 *. 552 can be used.

The close analogues of lineoinycin, i.e. compounds, in which R ^ H is an alkyl group in cis or trans with not more than 8 carbon atoms, R ^ is methyl or ethyl, R is an alkyl group with not is more than 8 carbon atoms, possess anti-bacterial ] £ properties; some are comparable to or similar to lincomycin superior to this, and can be used for the same purposes as lincomycin. Other analogs and isomers have similar anti-bacterial properties, but in lesser degree; these connections can lead to the same

25 -

008817/1888

BATH ORIGINAL

Purposes such as lincomycin are applied in cases in which the use of larger quantities does not cause any problems.

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

Example 1;

7-Ohlor-7-deoxylincomycin ^ ^- IvIethyl-7-chloro-6,7,8-trideoxy-6- (trans-1-methy1-A-propyl-L-2-pyrrolidinecarboxamido) -1-thio-L-threo -CL-D-galacto-octopyranoside ^

OH,

k & j

Eq

OIH

SGH,

XIX

A, the free base.

A suspension of 221.0 g (0.5 mol) of lincomycin hydrochloride in 5 liters of carbon tetrachloride was de under

- 26 -

0Ü9817 / 1888 BAD

162062Q

Nitrogen at 25 ⁰ G stirred well. Then 900 ml of thionyl chloride were added all at once, whereupon the mixture was stirred for a further 2 hours. During this time the solid dissolved and a clear solution was obtained. D '^ l Iieaktione; j; oini ^ Gh was loud for 2 hours; reflux was boiled, whereupon the ./arm source removed uric bticicstoff was introduced into the tea'nfoteinfarbsne Löluüc until the ivolbentempeivatur had fallen to 25 ° G. Then about 4 were "Liter of i'lii ^c -.. I" jkeit removed by vacuum distillation at a pot temperature of venig.r than 35 ° ^ The during you. '"Gesamuielt estillation abs · Heiiende yellow Festntoff • .vurde and dried , dr warde approximately 3G ^ al -kiethanol dissolved, the solution vmiie cooled to 25 ⁰ C, with vex · - dilute aqueous sodium hydroxide; (2-normal) adjusted to pH II, diluted with water to about 1200 ml and extracted well with ether. The thermal extracts are combined, washed with a small amount of water, dried over anhydrous magnesium sulfate and filtered. When lindanpfeu a _φ ίΕβΐ1ε of the combined ether extracts 7-GkIo ** - 7-deoxylincomycin (free base) was obtained as a yellow amorphous solid.

B. hydrochloride.

When adding Ohlor.vai-aeistoffgaa to the filtrate According to Part A, 7-chloro-7 * -deo :: yliiicomycin hydrochloride fell,

2 / -

009017/1888

BAD ORlQWAU

dcijat separated and recrystallized from ethanol and iithylacetate: i? rt wade. Was obtained in 32 / uiger yield dun weiüse crystalline ir-hydrochloride, solvaticiert with about 1 mol "Vasser ·

Analysis; For O ₁₈ H ₅₅ ClN ₂ O ₁ -U.HOl.HpO 0 45.18} H 7.57; 01 14.82; U 5.86; S 6.70 <H ₂ O: 3.77.

Found:

C 4'i, 70i H 7.65; 01 14.27; H 5.78; S 6.45; H ₂ O 3.85.

HoO
^? 7 _D '* ¹ ^ ⁰ (0-0.9858 g / 100 ml).

Activity: about 4- to 8-fold that of lincomycin

The anti-bacterial spectrum is the same as that of lincomycin.

Example 2:

7-0hlor-7 "* <ieoxylincomycin, Free Ease.

The procedure according to Example 1, Part A was repeated with the addition that dat. Ε: .iethylene chloride was used instead of ether as the extractant; The combined extracts were filtered and evaporated to dryness,! .lan received 64 ^ 7- * Ohlor-7-deoxylincoEiycin (free base • old amorphous solid. 15 / g of this pesticide was adsorbed on } Z g of silica gel in 1. dried and applied to a column filled with 15ΟΟ g of silica gel.

28 -

009817/18 8 8

SAO

162Q62Q

-was filled and eat a major chin; he was 7 »5 cm. the Column v / urde with a mixture of methanol and chloroform in a ratio of 1: 19 with portions of 200 ml, after 2 liteim Forerun, eluted. Fractions 26, 27 and 28 were merged and evaporated to dryness, yielding 1.04 g of essentially pure 7-chloro-7-deoxyl.1 neomycin in Form an amorphous solid with an anti-bacterial Spectrum similar to that of 7-chloro-7-deox ^ -lineomycin hydrochloride was the same as in Example 1.

Example 5t

7-chloro-7 * deoxylincomacin C ^

6- (trans-1-methyl- ^ - propyl-L-S-pyrrolidinecarboxamido) -1-thi-

L-threo-Ct-D-galacto-octopyranoside /

HO

01

OH

XX

- 29 -

009817/1888

BATH

so

A. Chlorine substitution »

A thin suspension of 1 g of lincomycin C hydrochloride

in 25 nil of carbon tetrachloride and 4-, 5 ^m l of thionyl chloride Lei 25 ° C was stirred for 2 hours under nitrogen. ..lan received a clear solution within about 15 minutes. The reaction mixture was then refluxed for 2 hours and evaporated to dryness in vacuo, yielding a yellow solid which was dried at 40 ° C. in vacuo for 18 hours. The product was then dissolved in about 15 ml of warm ethanol, made basic with sodium hydroxide as described in Example 1, and diluted to 300 ml with water. The aqueous solution was extracted five times with 100 ml of ether each time. The ether extracts were combined, dried over magnesium sulfate, filtered and saturated with hydrogen chloride gas, whereupon it was evaporated; a brown residue was obtained which, when recrystallized twice from a mixture of ethanol and ethyl acetate (dissolving in the smallest possible amount of ethanol and adding ethyl acetate until the onset of cloudiness), 200 mg of 7-chloro-7-deoxylincomycin 6-hydrochloride in " In the form of white crystals, diet.us product had the same activity and the same spectrum as the V-Ohlor-y-deoxylincomycin hydrochloride according to the example

0GS817 / 1888

ORIGINAL

Β «Manufacture of Lincomycin CU

Lincomycin G is obtained by reacting Lincomycin with ilthanthiol to form a diethyldithipacetal and heating up the peaction church in the present of p-iToluenesulfonic acid, or heating until it melts. The method is described in more detail below.

El. 6,3-Dideoxy-5- (tran8-l-met! Iyl ^ -propyl-L-2-pyrrolidinecarl3oxamido) -D-erythro-D-salacto-al iehydo-octose diet; hyl> dithioacetal.

^X 3

OU-

SSt

^{150 ml of concentrated hydrochloric acid and 50 ml of ethanethiol (cooled beforehand to 0} ° C.) were mixed in a 1 liter three-half flask, whereupon 15% lincomycin hydrochloric acid was added. After 5 hours of eating with a liquid stirrer, the Jemiüch was cured with an equal volume of water, '.

- 31 -

009ti 7/188 »

8AD OR1G »NAL

162062Q Sl

solve B (technical approach) was extracted; the extracts were discarded.

The main amount of acid was neutralized by carefully adding solid potassium hydroxide (100 g), while keeping the temperature of the well-stirred reaction mixture between 20 and 30 ° by cooling with acetone-dry ice. Solid potassium chloride was filtered off and washed with chloroform. Additional chloroform (ca 150 ml) was added to the filtrate and the mixture was adjusted to pH 10 by adding aqueous sodium hydroxide (2 normal) while stirring with a magnetic stirrer chloroform was layer separated, DIE aqueous layer was carefully. exträiiert it chloroform, the combined extracts were washed twice with water, then dried over anhydrous sodium sulfate · üeim removing the solvent at about 30 ⁰ G in vacuo a semisolid residue was obtained , which after crystallization from acetone 5 »4-lg 6,8-dideoxy-6- (trans-1-methyl-4-propyl-L-2-pyrrolidinecarboxamido) -D-erythiO-D-galacto-aldehydo-octose-diethyl Dithloacetal in the form of a colorless, flattened blame with a melting point of 130-132 ° C. Concentration of the mother liquors gave a further 1.50-6 of the product with a melting point of 129-131 ° C (total yield 6.91 g, ^ 2.4 fr).

- 32 -

00.117 / 1 888

. T62062Q

JJ

Analysis: Calculated for Gp-, H ^, ρ ^ ρ ^ 6 ^ΰ 2 ^:

O £ 2.25; H 8.77; JJ 5.81 ;. S 13.29 ^l / o, Found: O 52.58; H 8.71; S 5.93; 3 13.46 %.

E2o Gyklisicrung for Lincomycin Go

(a) One part Diäthyldithioacetal (gemäßs part Bl) and p-toluenesulfonic acid monohydrate were boiled in 25 parts of acetonitrile at reflux until substantial antibacterial effect of the product Wix ⁱ s was obtained. The reaction mixture was cooled and evaporated to dryness and then chromatographed on silica gel, using a mixed solvent of ethyl acetate, acetone and water in a ratio of 8: 5: 1. The 102 Ms fractions showed anti-bacterial activity. The fractions 105-125 were combined, evaporated to dryness and crystallized from acetone acidified with hydrochloric acid; Recrystallize by dissolving in water and adding acetone. File received to crystals of lincomycin G hydrochloride of melting point 149-153 ⁰ C.

(b) The üiethyldithioacetal according to part B1 was about

Heated to 260 G for 3 minutes, the smell of.

Ethyl mercaptan was perceived. The product yielded after Chr o / u at ο gra phi er en as described in part B2 (a), Lincomycin G

- 33 -

O & M M / 1 8 8 8 BATH

ü. Production of Lincomycin Q by fermentation.

Lincomycin G hydrochloride was prepared as follows:

Fomentation »

üj'in Üchrägnahrboden of ütreptomyces lincolnensis var. lincolnensis, KIiIiL 2936 has been inoculated for several 500 ml iSrlenmeyer-iColben verv.en-et, each of which 100 ml of the following iföhr medium:

Yeastolac

Glucose monohydrate

N-Z amines E.

Tap water to fill up

1 Xeastolac is a protein hydrolyzate made from yeast cells. 11 H-Z-Amin B is Sheffield's enzymatically degraded casein,

The pH - // ert of ITährmediums before sterilization was The fermentation was carried out for 2 days at 28 ⁰ G on a Gump-shaking machine with 250 rpm at 7 "3"

A 5 / "inoculum of the above medium (5 ml) each was placed in 30 500 ml Erlenmeyer flasks, which contained 100 ml of the following fermentation medium:

10 G 10 B. 0 1 Liter.

88 8

162062Q

15th era 40 Γ *
D. 20th E. 10 S. 20th ε S. e Ο , 5 ml Ι liter

Glucose monohydrate starch

Molasses

VZIlSOn ¹ B Pejton Liquor Ko. 159 Corn Steep Liquor Galcium Carbonate Bacon Oil

Tap water to fill up

1 Wilson ¹ s Peptone Liquor No. 159 is a preparation made from enzymatically hydrolyzed proteins of animal origin

At the time of the inoculation, DL-ixthionine was added in an amount giving a final concentrate of 2 g / ml.

The contents of the Schüttelkoll en.was worked up after 4 days of fermentation at 2P < O on a jump-shaking machine (250 rpm). The 'JehaIt var 200 i.! Ikrogramm per milliliter, in the C? Est against ia.lutea (Ee chieilunr see below) The total solids content fer fermentation vat tetiutr about ^ ug per liter *

235 liters of fermentation oil from finer ϊ · χ · mentation in Presence of DL-üthioiiin were teim pd-i / ert de ;: rcrjtionsendes filtiiert, by geleler, if necessary an i'Iterhilf

- 35 -

BATH

The mycelium cake was washed with water and then discarded. The filtered solution and wash water (275 liters) were stirred for 4-5 minutes with 12.5 kg of activated charcoal and 2.5 kg of diatomaceous earth. The mixture was then filtered and the The filtrate was discarded »The charcoal cake was washed with 60 liters of water, which was discarded» Then it was washed with 70 liters of 20% aqueous acetone, which was also discarded. The filter cake was then mixed twice with 100 liters each. aqueous acetone eluted. The jeluates were combined (215 liters) and the solution was concentrated to 18 liters. This concentrate was adjusted to pH 10.0 with 50% aqueous sodium hydroxide solution and then extracted three times with 20 liters of methylene chloride. The methylene chloride extracts were combined (60 liters) and then concentrated to give an oily preparation (7 §14 g) which contained lincomycin and lincomycin G in equal amounts as the free base. This preparation was dissolved in 200 ml of methylene chloride. The solution was filtered and then concentrated to dryness in vacuo. The residue was dissolved in 100 ml of 1N methanolic hydrogen chloride. The methanolic solution was then mixed with 3-2 ethers while stirring. Colorless, crude umcoisgreiii hydrochloride and lincomycin C hydrochloride precipitated out.

- 36 009817/188 «« ° «**« «.

was filtered and dried; Yield 7ί ^χΖ) - S » ^m i ^ a content of 9 ^ 0 micrograms per milligram against Sarcina lutea. (The test against Sarcina lutea is carried out on agar which is buffered with pH 7.0 phosphate buffer (0.1M) to a pH value of 6-3 ^ ibine volume unit (0.08 ml) of that to be tested The solution containing the material is placed on a 12.7 ml sample plate, which is then placed on an agar plate inoculated with the test organism give in about the same amount ·

7, Og crude lincomycin C hydrochloride were in 20 ml Water and 20 ml of butanol dissolved, whereupon the pH v / ert with I = A HCl set to 4.2 vjurde; the solution was in a countercurrent distributor distributed with 1000 transitions. By thin layer chromatography it could be determined that the Fractions in tubes 135 "to 190 contained lincomycin G · These fractions were combined and the solution became too concentrated in an aqueous mixture and freeze-drying subjected, giving 2.44 g of lincomycin C hydrochloride, at a level of 1400 micrograms per milligram against Sarcina lutea. 500 mg of this preparation was dissolved in 2 ml of water, 1 ml of methanol and 100 ml of icetone. The solution was passed through

- 37 -

000617/1983 BAD

162062Q Ji

Filtration clarified. The filtrate was then mixed with iLther, until the 'Veginnenden Crystallization' was thereafter the mixture was allowed to stand at room temperature for 1 hour. Crystalline lincomycin G hydrochloride (cube) was made by Decanting separated from the supernatant solution. The crystals vmrci.en recrystallized from 1 ml of water, 1 ml of methanol, 30 ml of acetone and 20 ml of iither, 250 ml of crystalline Lincomycin O-hydrochloriö Γ / ürfel) obtained. The above Admonished, supernatant liquid took 4 hours left at 5 G for a long time. Crystalline lincomycin fell in the process G-hydrochloride in the form of needles, which are filtered off; and have been dried; Yield 150 mg, melting point 151-157 ° C

Ρ «v / further procedure for Kerstellunp; by Lincomycin G.

8.85 S (0.02 mol) of lincomycin hydrochloride are dissolved in 20 ml of water, ^{cooled to 0} ° C. and, while stirring, 3.52 g (0.022 mol) of Erom are added over the course of one minute. Then 25 ml of ithanethiol was added and the mixture was stirred at 25 ° C. for 2 hours. The clear, colorless two-phase system (ethanethiol is relatively insoluble in water) was cooled in an iasbad and hydrogen chloride gas was passed in for about 5 minutes. The lower, aqueous phase turned red. The reaction mixture was then extracted three times with 100 ml of Skellysolve B each time, and aqueous sodium hydroxide solution was added to make the aqueous solution

-sä - οοβίΐ7 / ma .a «,

Adjust the phase to a pH of 11 * The basic phase was extracted well with chloroform, the chloroform extracts were washed with saturated sodium chloride solution, dried and evaporated in vacuo, giving 6 _{tons of} 2 g of a white solid, 4.8 g of this Solid * substance was chromatographed on 800 g of silica gel, proceeding according to the procedure described in Example 2, methanol-chloroform (II 7) was used as the solvent system. 58 were combined and evaporated to dryness, and the remaining solid was recrystallized from acetone to give 0.5 S · of a material identical to the diethyldithioacetal according to Part E1 · Fractions 65-75 were also combined, evaporated to dryness and dissolved in a mixture of 5 ml of methanol and 400 ml of diethyl ether · Bs · hydrogen chloride gas was added, and this separated out The white solid was collected "! recrystallization from aqueous acetone gave 0.5 S!" incomycin 0 hydrochloride, which was identical to the product according to part Q.

Em Other Alk5'l · -7-chloro-6 _t 7 ^ 8-tΓideoxy-6- (trans-l · --methyl-4 «■ propyl-If-2-pyrrolidine arboxamido) -l-thio-L-thr e oo> -D-galacto-octopyranoside "

If the ethanethiol in part B1 and part D of the above example is replaced by other alkyl mercaptans, e.g.

0098 17/1889

Propyl, butyl, pentyl, hexyl, iieptyl, octyl, honyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, isptadecyl, octadecyl, ITonadecyl or KLcosylmercaptane or isomeric forms thereof, alkyl mercaptans by Oyclo- as Oyclopropyl-, cyclobutyl, Oyclopentyl-, Gyclohex ^ ^r l-, Gyclolieptyl-, Oyclooctyl-, 2-methylcyclopentyl, 2,3-Dimetnylcyclobutyl-, 2-Methj ^r Icyclobutyl- or 3-Oyclopentylpropylmercaptane or by. Aralkylmer; aptane as zP Eenzyl-, phenethyl, 3 - ^ hCiIyIpI ¹ OPyI- or 1-Haphthylmethylfliercaptane so obtained nan the corresponding alkyl, cycloalkyl, and aralkyl-6,8-dideoxy-6- (trans-l-methyl - ⁷ J-propyl-L-2-pyrrolidinecarboxamido) -1-thio-D-erythro-O. —D— galacto — octopyijanocide which, when treated by the procedure of Part A, converts to the corresponding alkyl, cycloalkyl, and aralkyl-7-chloro-6,7ί ^p ^ trideoxy-6- (trans-1-methyl-4-propyl -L-2-pyrrolidincarboxamido) -l-thio-L-threo-0 * -D-galacto-octanopyranoside same spectrum of activity as lincomycin and same or greater activity.

- 40 -

0098 17/1888

ORIGINAL

Example 4; _r "*

Methyl-7-ch.lor-6,7,8-trideo3ry-6- (trans-1-alkyl-4-butyl-Ir-2-pyrrolidinecarboxamido) -l ^ thio-L-threo-c * -D-galacto -octopyrano " side.

GH, ■

01-

XXII

E, = ivlethyl or ethyl _e

Part A.

A suspension of 116 mg of methyl-6,8-dideoxy-6- (trans-1-ethyl-4-butyl-I / -2-pyrrolidinecarboxamido) -1-thio-D-erythro-aD-galacto ~ octopyranoside. in 3 ml of carbon tetrachloride and 0.7 ml of thionyl chloride was stirred at 25 ° 0 until a clear solution was obtained (approx. 13 minutes), whereupon the mixture was left to stand at 25 ° 0 for 2 hours. The reaction mixture was then refluxed for 2 hours and then evaporated to dryness in vacuo, giving a yellow

0098 17/1888

162062Q HZ

Solid was obtained, which was worked up as described in Example 3. However, the product did not crystallize, so that the solvents were evaporated off; this gave 10 mg of ethyl-7-chloro-6,7 _i ^R - "brideoxy-6- (trans-l-ethyl-A-butyl-L-2-pyrrolidincartoxamido) -l-thio-L-threo-O-D-galacto-octopyranoside hydrochloride as a buff-colored, amorphous solid, which against Gram-positive Ealferien the 8- activity of lincomycin and against Gram-negative the 16- to 64-fold activity of lincomycin.

When working with the cis epimers, one obtains the methyl-V-S-chloro-6,7 _r trideoxy-6- (cis-l-ethyl-4-butyl-L-2-pyrrolidincarboxamido) -l-thio- L-threo-CX-D-galacto-octopyranoside hydrochloride with the same anti-bacterial spectrum.

When the 1-IIethyl analogues are dislocated, one obtains Uethyl ^ -ehlor-ö ^ j ^ -trideoxy-e-Ccis- and trans-1-methyl-4-butyl-L-2-pyrrolidinecarboxamido) -l-thio-L-threo-0 ^ -D-galacto-octopyranoside hydrochloride.

The cis and trans jipimers used in the example above were made as follows:

B, 4-butylidene-I-carbobenzoxy-L-proline and its glyclo- hexylamins alζ.

19 g sodium hydride (in the form of a 5370 suspension in mineral oil) were added with 350 ml of dimethyl sulfoxide

-42- 009817/1888

162062Q 13

70-75 ° C. until the reaction was complete. (About 30 minutes) After cooling to 32 ° C., 16.2 g of eutyltriphenylphocphonium bromide were added and the resulting reaction mixture was stirred for 1 hour to complete reacting sicherzustel · len "Then, a solution of 26 g of 4-keto-l-carbobenzoy: / - L-proline in 100 ml of dimethylsulfoxide was added and the resulting mixture was 3 hours lan-v; heated to 70 ⁰ C. Then. The mixture was washed twice with 700 ml of ether each time, after which the zither was discarded after it had been back-extracted with 150 ml of 2.5% aqueous potassium bicarbonate solution The bicarbonate solutions were combined and acidified with 4-η hydrochloric acid. The acidified mixture was extracted four times with 500 ml of ether each time. The combined ether extracts were successively with 250 ml of water, three times with 250 ml of saturated aqueous iate each time rium bisulfite solution and washed with 250 ml of water and then dried over anhydrous sodium sulfate. When the solution was applied in vacuo, 24 g of an oily residue of 4-eutylidene-1-carbobenzoxy-L-proline were obtained.

This residue was dissolved in 31 el acetonitrile and mixed with 18 ml of dicyclohexylamine; then '.vuräe chilled. The crystals which separate out are puddled with acetonitrile washed and dried in vacuo, with uan 21 g

0 0 9 8 17/1888 BATH

(46.8 %) crystalline dicyclohexylamine salt of melting point 136-140 ⁰ G was obtained. After two recrystallizations from acetonitrile, 142-144 ⁰ Oj received nan an analytically pure sample with a melting point optical rotation _D ^ 7: -4 °

(c »» 0.99, ₃

Analysis: Calculated for

.G 7i, 86 | H 9.15; H 5.78.

Found: G 71.69; H 9.30 $ H 5.74.

10 g of the dicyclohexylamine salt were with ether shaken and excess 5% aqueous potassium hydroxide solution, "until no solid remained. The layers were separated and each was backwashed · The aqueous alkaline Layer was combined with the washing solution of the ether layer and acidified with 4N hydrochloric acid. The mixture was extracted repeatedly with ether, the extracts were combined, dried over sodium sulfate and evaporated in vacuo, whereby 6.3 g (93%) of 4-eutylidene-l-carbobenzoxy-L-proline in the form received an ul.

G. 4-Eutyl-1-carbobenzoxy-L-proline.

The OeI obtained according to Part E was in 200 ml Methanol over 2.1 g of 10% platinum on Dowex-1 at 2.8 Atmospheric hydrogen pressure hydrogenated. The catalyeator was filtered off and the filtrate was evaporated to give 6.3 g

- 44 -

0 0 9 8 17/1888

SAD ORIGINAL

162062Q «Γ

4-butyl-1-carbobenzoxy-aL-proline in the form of an oil. The product contained about 2 parts of cis -4-eutyl-1-carbobenzoxy-L-proline per part of trans-4-butyl-1-carbobenzQxy-L-prolino

Optionally, the hydrogenation of the 4-ylidene group to a later procedural stage, including the last procedural stage, to be moved «

If the butyltriphenylphosphonium bromide in part B is replaced by other substituted triphenylphosphonium bromides, the corresponding 4-alkyliclenes · ¹ ·, 4-cycloalkylidene and 4-ar-alkylidene-1-oarbobenzoxy-L-piOlins or the corresponding 4-alkyl are obtained -, 4-Oycloalkyl- and 4-Aralkylderivate »For example, with ethyl-, l- ^j ropyl-, isobutyl-, pentyl- and hexyltriphenylphosphonium bromides you get 4-ethylidene-l-carbobenzoxy-L-proline, 4-propylidene-l-carbobenzoxy» L-proline, 4-isobutylidene-1-carbobenzoxy-L-proline, 4-pentylidene-1-carbobenzo3cy-Ir-proline and 4-hexylidene-1-carbobenzoxy-L-proline and cis- and trans-4-ethyl-1-carbobenzoxy -L-proline, 4-propy 1-1-0arbobenzoxy-L-proline, 4-is'obutyl-1-carbobenzoxy-L-proline, 4-pentyl-1-carbobenzoxy-L-proline and 4-hexyl-1- carbobenzoxy-L-proline _e $.

De lvlethyl-6-amino-6, B-dideoxy-1-thio-D-srythro-ft. — D-galacto— octopyranoside (OL-MTL ·).

A solution of 40 g of lincomycin (U.S. Patent 3,086,912) in 20 ml of hydrozine hydrate (98-100%) was am

~ ^Λ3 ~ 0098 17/1888

BATH ORIGINAL

Reflux boiled; The hydrazine hydrate was removed in vacuo under nitrogen to steam bath temperature, leaving a residue of an astecic crystal mass. The residue was cooled, crosslinked with acetonitrile and the mixture was stirred until the crystals were suspended. They were then collected on a filter and washed with acetonitrile and iither. The yield of white, crystalline, Ok-I.I3Mj-free Ease after drying in the Yakuun at skin temperature was 21 g (84 "- / o) m equal volume of ethylene glycol dimethyl ether

The methyl-6-amino-6, ^p -dideoxy-l-thio-D-erythro-CJi-D-galacto-octopyranoside has a melting point of 225 ° -228 ° 0 _t optical rotation / g7j ⁵ : 4-276 ° ( c. - 0.768, '/ ater); pKa ": 7.45.

Analysis: Calculated for OqH, qNO, -S:

G 42.7; H 7.56; Vi 5.53; S 12.66. Found: 0 42.6; H 7.49; Ii 5.75; 3 12.38.

If lincomycin is replaced by other alkyl, cycloalkyl or aralkyl-6,8-dideo ^ .6- (trans-l-meth / l-4-propyl-L-2-pyrrolidinecarboxaiaido) -l-thio-D-erythro-

- 46 -

009817/1888 s _Ä0

162062Q 11

(XD-galacto-octopyranoside with the radicals ethyl, propyl, butyl, pentyl, hexyl, ileptyl, octyl, ifonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, JHlpjbadecyl, octadecyl or is, HonneradecylForms and Cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oyeloheptyl, cyclooctyl, 2-methylcyclopentyl, 2,3-dimethylcyclol> uiiylV ^ - ^M etEylOyclQbutyl or 3-Cyclopentylpropyl, benzyl, phenethyl, 2-methyl-methyl-propyl, so-pentyl or 1-methyl-propyl are so-pentyl or 1-thylmethylpropenten Alkyl, cycloalkyl and aralkyl-6-amino-6,8-dideoxy-1-thio-D-erythro-Ot-Bt-galacto-octopyranosides are obtained, for example, when replacing lincomycin with ethyl, propyl -, Butyl-, Pentyl- ^ Bder Hexyl-6,8-dideoxy-6- (tran.sl-methyl-4-propyl-L-2-pyrrolidinecarboxamido) -lthio-D-erytnro-ow-D-galacto-octopyranoside the propyl-, Butj'l-, Pentj'l- and hexyl-e-amino-e • bhio-p-erythro-Ot -D-galacto-oc topyranosid.

The procedure of Part A can also be applied to compounds under Part D, using of the hydrochloride or another salt of a strong acid, whereupon the resulting 7-chloro compound subject to the further procedural stages.

0 0 9 8 17/1888

BATH ORIGINAL

Methyl-e »8 ~ dideoxy-6- (l ~ carbobenzoxy-4-butyl-L-2-pyrrolidine-carboxamido ^ l-thio-D-erythro-ol-D-galacto · octopyranoside, free base «

XXIV

+ (X-MTL)

'0OH "V | / COMTL

To a solution of 6.3 g of 4-butyl-1-carbobenzoxy-L-proline (OeI according to Part E) in 175 ml of distilled acetonitrile, ^{cooled to 0-0} , were added 3.46 ml of triethylamine and then 3.34 ml Isobutyl chloroformate added. The mixture was ^{stirred at 0 0} O (£ 3 °) for 15 minutes, after which a solution of 6.2 g of IZTL-free phase according to Part C, in 85 ml V / ac.ser, was added; the reaction mixture was stirred at ^{0 0} G for half an hour and then at 25 ° C. for 1 hour. The reaction product was filtered off and dried, giving 4.57 g (37% 7%)! 8-dideoxy-6- (° l-carbobenzoxy-4-butyl-L-2-pyrrolidincartoxamido ^ -l-thio-D-eiythro-Oi-D-galacto-octopyranoside in the foir. Of the free base. urden

009817/188 8

- 48 -

162062Q H3

concentrated in vacuo, a further 4.25 S (35 %) of product being able to be obtained. On recrystallization from acetonitrile, diethyl-e ^ -dideoxy-6-Cl-carbobenzoxy-4-butyl-L-2-pyrrolidincarboxamido) -l-thio-D-erythro- CL- D-galacto-octopyranoside with a melting point of 194-196 ⁰ G obtained By further recrystallization from acetonitrile, an analytically pure sample with a melting point of 195-5-200 ° C. was obtained: f Hl ⁰ (c-0.98 MeOH) ₀

Analysis ; Calculated for Qo6®Ao ^ 2®8 ^ ^:

ö 57.75; H 7.46; if 5.13; S 5.93. ' Found: 0 57.58; H 7.16; H 5.50; S 6.07.

F. Methyl-6,8 ~ dideox ^ ^r -6- (4-butvl-L-2-pyrrolidincarboxamido) -l thio-D-err / thro-Q.-D-galacto-octopyranosid-hydrochloxid *

■>

OOM ^r J? L

Solution of 7.8 g of methyl-6,8-dideoxy-6- (l-carbobenzoxy-4-butyl-L-2-pyrrolidinecarboxamido) -l-thio-D-erythro-iX-D-galacto-octopyranoside, made in part E,

0098 17/1888 - 49 -

BATH ORIGINAL

in 20% ml of methanol was over 2 g of 10% i-aliadium-carbon oil catalyst at 2.0 atmospheres ./assert toff 1? Hours lung shaken. The catalyst ^ urde then filtered off and the solution was in the Wx J.um a & c. en ^ t. The residue was dissolved in a mixture of 20 ml of acetone and 20 ml of JusEer and acidified with 6N hydrochloric acid. Dilute with 4 Yolumenteilcn acetone was -.! Ethyl 6,8-dide oxy-6- (4-butj ^r IL-2-pyrrolidincarboxamido) -1-thio-D-eiythro-OW-D-galacto-octopyranosid hydrochloride from which was filtered off and dried. The crystals obtained in this way were dried at 55 3 in vacuo; Yield 4.7 S * melting point 188-194 ^{0 0} ₀ The analytically pure sample obtained by recrystallization from acetone had a melting point of 197-199 ° C; / 07Jp: 150 ° (, Va ^ ser, c = 0.89).

Analysis: Calculated for CngE ^ i ^ Ogü.H'Jl:

G 48.80; H 7.96; Il 6.32; S 7.24. Found: (corrected for 5.54% Japser)

G 48.58} H 8.19; 1Ϊ 6.04; S 7.36.

That material possessed 8% of the anti-bacterial activity of lincomycin against Selutea.

If the ß-MTL is replaced by other alkyl, cycloalkyl or aralkyl-6-amino-6,8-dideoxy-1-thio-D-erythro- (jL-D-galacto-octanopyranoside, in which the alkyl radical is ethyl,

- 50 -

0098 17/1888

BAD ORJGIN _AL

Propyl, butyl, pentyl, hexyl, rieptyl, octyl, iionyl, decyl, undecyl, dodecyl, tridecyl, tctradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, ITonudecyl odor licosyl or their isomeric forms, the oycloalkyl, cyclopentyl, Oyclohex; ^r l, üyclohepty L, Oyclooctyl, 2-Methylcyclopcntyl, 2,3 -Dime thy Icy el obuty 1, 2 -: Ie thy lcyc Ιο-butyl or 3-Cyclopentylpropyl and the aralkyl radical benzyl, phenethyl, 3-Pheiiylpropyl or 1- Naphthylmethyl i :: t, εφίθΓαβη the corresponding alkyl, cycloalkyl and aralkyl-6, S-dideoxy-6- (l-carbobenzoxy-4-butyl-L - ^ - pyrrolidinecarboxamido) -l-thio-D-erythro -eL-D-galacto-octoporanoside and alkyl-, Gycloalkj'l- as well as aralkyl-6,8-dideoxy ~ 6- (4- "butyl-L-2-pyrrolidincarboxamido) -l-thio-D-erythro-a - D-galacto-octopyranoside, for example, a replacement of OL-IIITL with ethyl-, Iropyl-, eutyl-, l-entyl- and iiexyl-6-amino-6,8-dideoxy-1-thio-D is obtained - erythro - ^ - D-galacto-octopyranoside ethyl-6,3-dideoxy-6- (l-carbobenEoxy- ^z f-butyl-L.2-pyrrolidinecarboxamido) -D-erythro-oil-D-galacto-octopyranoside , Propyl-6,8-dideoxy-6-Cl-carbobenzoxj - ^ - butyl-L-2-pyrrolidincarboxamido) -1-thio-D-erythro-Ä-D-galac to-oc topyranosi d, 2uty1-6,8 -dideoxy-6- (1-carbobenzoxy-4-butyl-L-2-pyrrolia-carboxa mido) -l-thio-D-erythro-ot-D-galacto-octopyranoside, rentyl-6,8-dideox ,. -6- (1-carbobenzoxy— 4-tutyl-L— 2-pyrrolidine-

009817/188 8

BATH ORIGINAL

carboxamido) -l-thio-D ~ erythro ~ ol-I) - galacto-octopyranoside, hexyl-6,8-dideo2cy-6- (l-carbobenzoxy-4-'butyl-L-2-pyrrolidine- · Carboxaraido) -l-thio-D-erythro-fll -D-galacto-octopyranoside, ethyl-6,8-dideo: xy ~ 6- (4-butyl_L-2-pyrrolidincarboxamido) -l-thio-D-erythro -iX-D-galacto-octopyranoside, prop ,; 1-6,8-dideoxy-6 ~ (4-butyl ^ L-2-pyrrolidinecarboxamido) ~ l-thio-D ~ erythro- (XD-galacto-octopyranoside, eutyl-6,8-dideoxy-6- (4- butyl-L-2-pyriOlidinecarbo>: amido) -l-thio-D-crythro- A D-galacto-octopyranoi.id, ientyl-6,8-dideoxy-6- (4-butyl-L-2-j. yrrolidincarboxamido) -l-thio_D-erythro- (X -D-galactooctopyranosid, üexyl-6,8-dideoxj ^r - «6 - (4-buty 1-L-2-i; yrrolidin- 'carlo :: amid ^ - l-thio-D-er3 ^T thro- ft -D-galacto-octopyranoside.

It replaces 4-lutyl-1-cartoberizoxy-L-pi'oline, other 4-alkyl-1-carbobenzoxy-L-proline, which are used as. Ethyl-, propyl-, eutyl-, 2: entyl-, Hgx., 1-, III.pt ^ l-, octyl-, ilonyl-, decyl-, uii'jocyl-, uodecyl-, yridecyl-, tetradncyl-, Pentadecyl, hexadecyl, heptadecyl, üctadecyl-, -ionadecyl- or Eicocylreste or their isomeric forms, or by ^ -Gycloal ^ llc-irbobenzoxy-L-Moline with Gyclopropyl, 'Jyclotat3 ^/ 1, G./clopentyl, Gyclohexj ^T 1, cycloheptyl, cyclooctyl _T 2-meth; / lc; 'clopent: 7l _i 2,3-dimethylcyclobutyl, 4-methylcyclobutyl and 3- ^ yclopr.ntylpropyl in the 4-position, furthermore

- 52 -

by 4-aralkyl-l-earbbbenzoxy-L-proline in which the 4-aralkyl is benzyl, Phonäthyl, 3-Phe nj 1 propyl or 1-iTaphthylmethyl may be, so to obtain the corresponding alkyl, cycloalkyl and aralkyl e ^ -dideoxy-o-Cl-carbabenzoxy-4-alkyl, ^ -cycloalkyl and 4-aralkyl-L-2-pyrrolidincar "boxamido) -l-thio-D-erythro-Ä-D-galacto-octopyrano3ide so; ie alkyl -, Cycloalkyl- and aralkyl-6- (4-alkyl, 4-cycloalkyl, 4-aralkyl-L-2-pyrrolidincar'boxamido) -l-thiO'-D-erythro- (Ä - D-galacto-octopyranoside «

Obtained, for example the replacement of ^z (--Butyll-carbobenzoxy-L-proline by 4-methyl, 4-Athj ^r l, 4-propyl, 4-pentyl or _4-hexyl-l-carbobenzox ^ ^r - L-proline methyl, ethyl, propyl, butyl, pentyl and hexyl-oja-dideoxs ¹ --6 - ^^ (l-carbobentoxy-4-methyl-L-2- «pyrrolidinecarboxaiaido) -1-thio —D-erythro — O -D-galacto-octopyranoside; methyl, ethyl, propyl, eutyl, pentyl and hexyl-6, S-dideoxy-6- (l-carbobenifoxy-4-ethyl-L- 2-pyrrolidincarbox amido) -1-thio-D-erythro-0.-D-galacto-octopyranoside; methyl, ethyl, propyl, butyl, pentyl and hexyl-6, S-dideoxy-6- (l -carbobonzoxy-4-propyl-L-2-pyrrolidine-carboxamido) -l-thio-D-erythro- ^ - D-galacto-octopyranosidj methyl-, ethyl-, propyl-, butyl-, pentyl- and hexyl-eje- dideoxy-ö-Cl-carbobenzoxy- ^ l-pentyl-L ^ -pyrrolidincarboxamido) - 1-thio-D-erythro-o * - D * -

-53 -

0098 17/188 8

BATH ORIGINAL

162062Q SH

galacto-octopyranosidj methyl, ethyl, propyl, putyl, Perityl- and tiexyl-6,8-dideoxy-6- (l ~ carbobenzoxy-4-hexyl-L- 2-py rrolidincarboxamido ^ l-thio-D-erythr ο- <λ-D-galactooctopyranoside; Methyl, ethyl, propyl, butyl, pentyl and hexyl-6, 8-dideoxy-6- (4- methyl-L-2-pyrrolidinecarboxamido) -l-thio-D-erythro-CL -D-galacto-octopyr? Methyl, ethyl, piopyl, eutyl, pentyl and hexyl-6,8-dideoxy-6- (4-ethyl-L-2-pyrrolidine-cartoxamido) -l-thio-D-erythro-O. -D-galacto-octopyranoside; Methyl, ethyl, Propyl, butyl, pentyl and hexyl-6,8-dideoxy-6- (4-propyl-L-2-pyrrolidinecarboxamido) -l-thio-D-erythro-Ä-D-galacto- octopyranosidj methyl, ethyl, propyl, eutyl, pentyl and hexyl-6,8-dideoxy-6- (4-pentyl-L-2-pyrrolidinecarboxamido) -1-thio-D-erythro-OL-D-galacto-octopyranoside and methyl, ethyl, propyl, eutyl, pentyl and liexyl-6, S-dideoxy-6- (4-hexyl-L-2-pyrrolidinecarboxamido) -l-thio-D-erythro-oil -D-galacto-octopyranoside

If desired, the 1-carbobenzoxy compounds prepared in Part E can be chlorinated by the procedure of Part A and the resulting 7-chloride compound can be treated through the further steps of the process to remove the 1-carbobenzoxy group and substitute the proline nitrogen fs

009817/188 8

SS

Gl,

carboxamido) ~ l-thio ~ 3) ~ erythro-Oi - D-ealacto-octopyranosidhydj-o chloride.

Gn ₃ XZiVIII

IiOl , ν .HOl

GOMTL

'' JOMT]

A solution of 2, Cg: .Iethyl-6, 8-dideoxy-6- (A-butyl-L-2-pyrrolidinecarboxamido) -l-thio-D-erythro-O ^ -D-galacto-octopyranoside-hydrochloride according to Toil and F '? 2.0 ml of 3 ^ pc alcohol in 150 ml of formalin was Methanoi-üb τ 500 mg' lOjcSigem palladium-carbon -iiataly cat or at 2.8 atmospheres .Vaesorttoff 5, ^"c hours lan - g". -chvttelt. li-jch removal of the catalyst and removal of the solvent ε ia VakuxT. partially crystalline ethyl-6,8-dideox5 ^r -6- (l-met} i3. ^r l-4 -butz 1-L-2-pyrrole! dincarboxamido) -l-tiiio-D-erythro-iX-D-galacto-octopyrunoside hydrochloride obtained by thin-layer chromatography on silica gel using a mixture of ethyl acetate, acetone and water (8: 4 : 1) for elution: and a potassium peiiaanganate solution for pectification as main-

009817/1888

BATH ORIGINAL

was recognized as consisting of 2 materials ", namely the cis and trans epimers of methyl-6,8-dldeoxy-6- (l-methyl-4-butyl-L-2-py rrolidincart> oramido) * - l-thio-D * - erythro-OIL-D-galacto-octopyranoside hydrochloride in the ratio of about 3 J 2 *

Q2> Separation of the ice and trana-f form by. Chromatography »

The obtained according to Part Gl methyl-6 _t 8-dideoxy-6- (1-methyl - ^ - butyl-L ^ -pyrrolidine-carboxamido) -l-thio-D-erythro- <l -D-galacto-octopyranoside- hydrochloride were dissolved in a mixture of methanol and methylene chloride (1.Jl) and treated with 1.5 ml of triethylamine. 7 g of silica gel were added to this solution and the solvent was evaporated in vacuo. The antibiotic deposited on the silica gel was subjected to a chromatographic test Abandoned column containing 200 g of silica gel soaked in a solution of ethyl acetate, acetone and water in a ratio of 8: 4: 1. The column was developed by eluting with the same solvent system, with portions of 20 ml each being collected. By thin-layer chromatographic examination of the individual c. Fractions it was found that fractions 31-38 (310 mg) consisted of weekly pure trans-epimers,

- 56 - ·

009817/1888

the fractions 49-7 ^ (32 mg) from essentially pure cis epimers, and fractions 39-4-8 from a mixture of epimers. The latter could be separated * The individual Bpimeren were dissolved dilute hydrochloric acid in little drops, after which the hydrochloride was precipitated by addition of acetone · In this manner, there was obtained 50 by repeatedly Ohromatographieren mg methyl 6 _t 8 "dideoxy> p-6- (trans —1-methyl-4 ~ buty1-L-2-pyrrοlidinecarboxamidο) -1-thio-D-erythro- ©. «D-galacto-octopyranoside hydrochloride with a melting point of 135-137 ° Ο and about 150 mg methyl-6.8- dideoxy «6- (cis» -l «-methyl-4-butyl ~ II-2-pyrrolidinecarboxamidoO-l-'thio-D-erythro-Ä-'D ^ 'galacto-octopyranoside hydrochloride with a softening point of 105 °, melting point 175-185 ° C.

The trans epimers melted after recrystallization from the same solvent at 139-141 ⁰ O '

AiIaIySe ₁₁ S ₁ Calculated for G ^ gH, ^ NgOgS ^ HGl s

G 49 ₉ 93§ HS ₃ 16ι Έ 6 ₈ 13 | B-7.02. Found? (corrected tÜT 4.07% HgO) -ί-

G 40 ₉ 81j H 8.54ι N 6.49 | S 6.67.

From the ice "epimers was obtained upon recrystallization a product Toii softening point 108 ⁰ G ₉ * melting point 189 ⁰ G (solvate). '

if / S && β

BATH ORIGINAL

Analysis ι Found: (corrected for A-, 95 % water):

G 50.27; H 9.0Oj N 6.05; S 6.65.

The trans-epimer was about 2.2 times as active lincomycin (against S- * lutea), about twice as active in nutrient * - "Broth-Diluent, and 2.5 times as active with S. aureus infected mice

The cis epimer was about 1/2 "to 1/3 as active as the trans-epimer and about the same as lincomycin.

. Methy 1-6,8-aideoxy-6- (l-ethyl-4-butyl-L-2-p: / rrolidine · " oar "boxamido) ~ l -!" th.io - D-erythro ^ Q .- »D-galacto-octopyranoside> * hydrochloride.

A mixture of 2.0 g of methyl 6,8-deoxy-6- (4- «butyl-Ir-2-pyrrolidincar" boxamido) -l-thio-D- «erythro-CL-D-galacto-octopyranoeid hydrochloride according to Part P, 1.5 ml of acetaldehyde and 150 mg 10% palladium-carbon in 150 ml of methanol was shaken at 2.5 atmospheres of hydrogen pressure for 5 »5 hours. The catalyst was at-filtered and one obtained Residue, mainly from the cis and trans epimers of methyl-6,8 "dide oxy ~ 6" (1-ethyl-ή-butyl-L-2-pyrrolidine carl DOxamido) -l-thio-D “erythro-OIL-D-galacto-octopyranoside hydrochloride duration*

- 58 -

009817/1888.

⁰ ORi _QiNAL

H2. Separation of the epimers.

The above mixture of epimers (2 g) was added to 200 g ßilikagel chromatographed, whereby one for the elution a solvent system of ethyl acetate, acetone and water (8 »4il) used« · Fractions 33-42 consisted of pure trans epimers (by thin-film chromatography) and hurdles united $ Factions 49 and 64 passed of essentially pure cis epimers, they also became united. The factions '43-48 consisted of one Mixture of epimers obtained by repeated chromatography could be separated. Each of the epimers was in v.-enigen Drops of dilute hydrochloric acid dissolved, and the crystalline hydrochloride was diluted with a large volume lther failed.

The fraction from crude trane epimers v; og 415 mg; 340 mg (15.4%) of crystalline methyl-6 _# 8-dideoxy-6- (trans-l-ethyl - ^ - butyl-L-2-pyrrolidinecarboxamido) -l-thio- «D-erythro- 0.-D-galacto-octopyranosidhydrochlorid of melting point 144-151 ⁰ G. recrystallization from acetone dilute the Schiaelzi -onkt rose to 148-151 ° G.

The fraction of the ice epimer weighed 645 mg and gave 300 mg (I ² Sl? ») Of crystalline methyl-6,8-dideoxy-6-

- 59 -

0 0 9 8 17/1888

BATH ORIGINAL

(cis-l-ethyl-4-'butyl-L-2-pyrrolidinecarboxamido) -l-thio-D-erythro-Ct-D-galacto-octopyranoside hydrochloride of melting point 135-JL39 ° G · When recrystallizing from dilute Acetone gave crystals of melting point

The trans epimer showed the 1-1.2-fold activity of lincomycin against S. lutea, the 2-4-fold activity of Lincomycin against grievous organisms and those ain3 estene 8-fold activity of lincomycin «against gram negatives Organisms that was in the mouse test against S. aureus trans epimers about twice as active as lincomycin} that cis-Jäpimere showed about 1/2 the activity of dee trans epimers.

The separation of cis and trans isomers is not absolutely necessary that the epimer mixture of the 7-Ohlor- « derivatives can also be used as such.However, it is desirable to keep the content of trans isomers high, since this isomer is the more active form. When carrying out the process, mixtures of epimers can be used with a ratio of trans to cis epimer from 3ti to 1: 5 can be obtained. If formaldehyde and acetaldehyde are replaced according to parts G and H by other oxo compounds of the formula R ^ RcCO, z.t. by propionaldehyde, acetone, butyraldehyde, Isobutyl methyl ketone, benzaldehyde, phenylacetaldehyde,

- 60 -

009817/1888

Iydrossimtaldehyd, Aeeijpphenan, Propiophenon, Butyrophenon, 3-Methyl-4-phenyl-2 - 'butaneQn, 2-Methyl-5-Phenyi-3- pentanoen, 3-Cyclopentanpropionaldehyd, CyclohexanaGetaldehyd, Cyclo-heptanarboxaldehyde, 2.2 -Dimethylcyclopropanacetaldehyd, 2,2-Dimetl3ylcyclopropylmethylketoa _J Cyclopentylmethylketon, GyGloTDu. "bylmeti3ylketon, Oyclobutanon, cyclohexanone or 4-Methylcyalohearanon using the appropriate alkyl, cycloalkyl or aralkyl · = 6, 8" dideoxy-6- (4-alkyl , 4-octoalkyl or 4 "aralkyl-L - ^^ pyrrolldincarTDoxamido) - 1-thio-D-erythro =" ®. "" D ^ galacto-oGtopyranoside, the corresponding alkyl- ₉ cycloalkyl- and aralkyl-6 are obtained , 8-dideoxy-6- (1 = R ^ HcöH «^ =« alkyl-, 4 «sycloalkyl- and 4-aralkyl« L · -2-pyrrolidine · »© ar · boxamido) -l-thio-D« erythro ~ A * -D-galactoootopyranoside, which "on treatment with thionyl chloride according to the process according to! Part A to the corresponding alkyl, cycloalkyl" and aralkyl ~ ₉ 7-QhIOr-O, 7 "8-trideoxy-6- (lH ^ HcCH-il-alkyl-ft ^ cycloalkyl «and 4-aralkyl-L ~ 2-p yrrolidincarboxamido) - "l-thio" L-threo- Ok-D-galacto-octopyranoside, in which ILR ₁ -OH- propyl, isopropyl, butyl or 4-methyl, 2-peirtyl, benzyl, phenethyl, 3-phenylpropyl, 1-phenylethyl, 1-phenylpropyl, 1-phenylbutyl;, 3-methyl-4-phenyl- »2« "butyl or 2-» methyl-5 * -phenyl- "3-peiityl,

- 61 -

009817/1888

BATH ORIGINAL

3-Oyclopentylpropyl, 2-C3 / clohexylethyl, cycloheptylmethyl, 2- (2,2-iDimethylcyclopropyl) ethyl, 1- (2,2-dimethylcyclopropyl) ethyl, 1-cyclopentylethyl, 1-cyclobutylethyl, cyclobutyl, cyclohexyl or 4- methylcyclohexyl · V'erv; ends iaan Foimuldehyd, acetaldehyde or other alkenols such, e »xropionaldehyd, butyraldehyde, valeraldehyde or Gaproaldehyd together with an alkyl-6 _t S"dideoxy'-6- (4-alkyl-L-2 -pyrrolidincarl) oxamido) -1-thio-D-eryfchro-fll -D-galacto-octopyranoside, in which alkyl and 4-alkyl, methyl, ityl, propyl, eutyl, pentyl, or hexyl are preferred starting compounds of the formula

XXlX

obtain,

in which X is a hydroxyl group and R, KR, and Ea are alkyl radicals with not more than 6 carbon atoms and

- 62 ~

009817/1888

SAD ORiGiNAL

preferably with no more than 12 carbon atoms represent in the overall aggregate which have the configuration D-erythro, which when treated with thionyl chloride lead to compounds of the formula XXIX, in which X is chlorine, E, HR, and R »alkyl have the meaning given above, where di · configuration is probably L-threo. The 6,8-dideoxy compounds of the formula XXIX are active anti-bacterial. Means that can be compared to lincomycin. the 7-chloro, 6,7,8-trideoxy compounds of the formula XXIX the same spectrum of anti-bacterial effectiveness, but are much less active · representatives of compounds of the formula XXIX with X ■ chlorine, of which assumed to have the L-threo configuration are shown in the following table:

-63 -

009 817/188 8

BATH ORIGINAL Table I. Compounds of the formulas XXIX with X - chlorine »

(7-Ghlor-7-de oxy-
lincomycin) E. HR ₁ 3 4A (7-chloro-7-deoxy-
allolincomycin) methyl trans-ethyl methyl 4 B (7-chloro-7-deoxy-
iincomycin E) methyl cis-ethyl methyl 40 allolincomycin E) methyl trans-Propy1 methyl 4D Methyl ' cis propyl methyl 4E methyl trans-propyl ethyl 4F methyl ciss-propyl ethyl 4G ethyl trans-propyl methyl 4H ethyl cis-piopyl methyl 41 methyl trans- "butyl methyl 4Y Methyl ci- "butyl methyl 4K methyl trans -propyl ethyl 4L methyl cis propyl Ätfcyl 4M ethyl trans-propyl ethyl 4N ethyl .ei ε-Pr opyl ethyl 40 methyl trans-butyl £ thyl 4P methyl cis-eutyl ethyl 4Q methyl trans-pentyl methyl 4R methyl cis-pentyl methyl 4S ethyl trans-butyl Ät ^l -vl 4T ethyl cis-butyl ethyl 4U methyl trans-pentyl ethyl 4V methyl cie-pentyl 4W Ethyl trans-pentyl

- 64 -

009817/1888

$ s

1620 ^ 20

AtjJs? 3 * <& is-? j? ej $ fcyl. ; trans-pentyl . methyl · 4Ϊ methyl trans-HesQrl cis-pentyl methyl 4Z " Met, ayl ois-Sexyl trans-pentyl methyl Biatyl trans-Propy. l cis-pentyl -.."Methyl 4AB Butyl cis ^ propyl methyl 4AG ethyl trans-pentyl lthyl 4AD lthyl cis-pentyl lthyl 4Al Butyl trans-butyl lthyl M-HB Eutyl ois »butyl lthyl 4AG Butyl trans-Peaty1 methyl 4AH Butyl cis-pentyl methyl 4AI Cyclohexyl trans-propyl methyl 4AJ Cyclohexyl cis propyl. . methyl 4AK Butyl lthyl 4AL Butyl lthyl ' 4AM Pentyl lthyl 4Old Pentyl lthyl

Intermediate products for the preparation of the above compounds are compounds with (1) X »hydroxyl | (2) E ^ »hydrogen;

(5) X - hydroxyl and R ₅ ■ - hydrogen; (4) X = hydroxyl and Rz - carbobenzoxy; (5) X. » Chlorine and E ^ - · Garbobenzoxy;

(6) X - hydroxyl j K ^ ■ Oarbobenssoxy and HE ₁ and H in the 4-position are replaced by the xlidene group E ^; (7) X chlorine, E, "carbobenzoxy and HR, and H in the 4-position. Are replaced by the ylidene group E-, Jf (8) X - hydroxyl, R * hydrogen, and HE ^ and H in the 4" position replaced by. -

0098.17 / 1888

BATH ORIGINAL

by the Xlldengruppe R ₁ J (9) X * · chlorine, R, - »hydrogen hydrogen, and HR ₁ and H in the 4-position are replaced, by the X group R-jj (10) X - hydroxyl, ur-d 1 'H ₁ and E in 4-D are replaced by the ylidene group R ₁ I and (11) X "chlorine and HR ₁ and H" in 4 "* position is replaced by the T group R ₁ , where z \ i h ^ ach ^ ön iac that "with X - Hy the configuration is D-erytcxo; and" with X ■ chlorine, the configuration is probably L-threo ·

If lincomycin according to Part A of Example 4 is replaced by an alkyl, cycloalkyl or aralkyl ^ -amino 6,8-dideoxy * 3. «Thio-D-ery thro-CJ - D-gal act o-octopyranoside, compounds of the formula are obtained in this way

XXX

BR

CH

in which ethyl, propyl, butyl, peatyl, kexjl, heptyl, Octyl, nonyl, decyl, t-indecyl, dodecyl, tridecyl, tetradecyl, Pentadecyl, hexadecyl, heptadecyl, octadecyl, ITonadecyl or Bicosyl or an isomeric 3? Form thereof, Gyclopropyl, Cyclobutyl, Gyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl,

- 66 -

1888

SAD

620620

S-Methylcyelopentyl, 2 ₀ 3-Bimetfayleyelobutyl, 2-Iiethylcyelobutyl or 3 «^ yelopentylpropyl, Benzyl, Phenäthyl, 3-Phenylpropyl or 1-Naphthylmetnyliet _? which can be acylated by the process of Example 4 to form the corresponding ^ -Cklor ^ -de oxylineomycin "analogues" for example of the compounds of the formula XH3C, and their intermediates

Example 5ι

Methyl-7-chloro-6,7 * 8-trideosy-6 «(trans-1-methyl - ^ - propyl-Ιί-2-pyrrolidinoarboxaiaido) -1-thio-De ry thro -CX -D- gal ac t ooot opyranossid (7- * Gh2.or-7 " ^| deoxy ~ epilinc omycin)

A · J. ^ - O-Isopropylidei-lineoinycin

OK _x

ΣΧΧί

V. 67 -

009817/1888

BATH

OO

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 vigorous stirring and exclusion of moisture. The mixture is stirred at room temperature for about 1 hour, whereupon 100 ml anhydrous ether can be added; Stirring is continued for 1/2 hour in an ice bath. The mixture is then filtered and the solid is ^{dried in vacuo at 50 °} C.; Yield 13.35 g (85.5%) of 3-4O-isopropyl -ldenlincomycin ptoluenesulfonate. A further 1.15 g (7.4%) can be obtained from the mother liquors by adding 350 ml of anhydrous ether and cooling the solution for one hour. The 1.5 g thus obtained are suspended in 200 ml of ether and shaken vigorously with 125 ml of 5% potassium bicarbonate solution. The aqueous layer is back-extracted twice with 100 ml of ether each time. The ether extracts are washed with 50 ml of saturated sodium chloride solution and then filtered through anhydrous sodium sulfate. The ether is distilled off in vacuo, with 7 , 9 g (73.1%) 3, ^ - O-isopropylidene lincomycin remain, which is dissolved in 25 ml of ethyl acetate, whereupon the concentrate is concentrated to 10 to 15 ml. The concentrate is left to stand at room temperature for several weeks and then cooled over night The crystals are filtered off from the solution

- 68 -

00981 7/1888

and washed with a little cold ethyl acetate; Yield ^ ■ »55 g (4-2.2%) of 3» 4 ⁱ ~ 0 - = - 'Isopropylidenelineomycin with a melting point of 126-128 ⁰ Gj optical rotation ^? Ψ s 101-102 ° (c - 1 methylene chloride),

B. 7-3> ehydro-3, ^z t-0-isopropylidene lincomycin.

12 g (excess) chromium oxide were added to a solution of 6 g (0.0135 mol) of isopropylidene lincomycin in 75 ml of pyridine 250 ml of ethyl ether and ethyl acetate contained. The mixture was then filtered and concentrated to a syrup (8.4 g). This syrup was distributed in a countercurrent distribution system with 500 passages using the solvent system water / ethyl acetate / ethanol / cyelohexane (IsIslsi). 7-Dehydro-3 »4-0-isopropylidene-lincomycin was isolated from tubes 330-380, K. 2.45

Analysis; Calculated for ö ^^ H ^ glipOgSi

G 56 ₉ 72 | H 8.16 $ -B "6 ₉ 30 | S 7.2Io . Found! 0 56.375 H 7 _S 62; Ή 6 ₉ 51j S 6.84 *

Oe 3 ₉ 4 "0" .Isopropylidene- = ep ± lincoiaycin _e

To 1.6 g Grai g-re INEM 7 ^e dehydro 3 ₉ 4 '= <) ^ isopropylidene-lincomycin in 75 ml of methanol were 400 mg Matrium = "boriiydrid eye sat ifach 1 ₉ 5 hours dia solution

69 »0 0 9 8 17/1888

BAD OBIGiNAL

25 ml of water were added to the residue, whereupon it was extracted three times with 25 ^ l of methylene chloride each time. The extract was washed back with 15 ml of water, then dried over magnesium chloride and evaporated to dryness residue was g of 1.4 in a countercurrent ^ distribution system with 5oC transitions distributed using a solution Sittel system of water: ethyl acetate ethanol: G3dohexan (Islilsl), and it was a einsige tip "which corresponded to the theoretical" Yert in K m. 1.05 observed. The material from tubes 240 to 280 was isolated in the form of a syrup »

Analyzes Calculated for Cp] H ^ gNgOgSs

C, 56.47i, H, 8.58; N 6.2? I 3 7.18. Found «0 56.24j H $ 8.54 N 6.13; S 7 * 01.

By thin-layer chromatography it was determined that this material consists of two substances, namely the 3 ^ -Q-Isopropylidene-lincomycin and the 3,4-Q-Isopropylidenepilincomycin existed, which hiked about a & slower.

D. Epiliiicomysiße

The syrup according to rope G was for 5 hours; held at room temperature in a solution which contained 60 ml of 0.25 N hydrochloric acid and 40 ml of ethanol. The mixture was kept at ^{0 0} Q for 4 days.

70- 009817/188 8

"Bicarbonate neutralized, concentrated to 25 μl and extracted with chloroform. The extract was treated with a little water and dried over magnesium sulfate and then evaporated to dryness. Thin-layer chromatography of the residue revealed two substances, both of which were active against Solutea. The residue became." ( "" type described 393625 Patent 2 * packed column (35 x 1 synthetic silicate in U _e S) »8 cm) eluted at a with Flosisil gradually, wherein the solvent continuously from 100% Skellysolve e (technical hexane) in 100% The total volume of solvent was 5000 ml. In this way, the compounds in the case of len were separated. '

Fraction 1: "tubes 53-65 (fractions of 40 ml each) Epiliaeoiayaiiu
Tested ^ -50 mcg / ralo.
Analysis: Calculated for ö, gH ^ -i ₂ ^ 6 ^Ss *

G 5C, 92i H 8.55i Ii 6.60; S 7.56, Gefunäens 3 50.19; H 7 »91 | N 6.05? S 6.42.

Fraction Hi tubes 73-10 *} -. Lincomycin Test value: 950 mc g / mg.

0 0 9817/1888

BAp ORSGiNAL

162062Q

σι

NH

HO / -D

XXXII

Kqh J

\ ^ SCH

OH

0 »85 S 7-epilincomycin hydrochloride were suspended in 17 ml of carbon tetrachloride. Then, 4.5 of thionyl chloride were added and the reaction mixture was stirred for minutes "at 25 ⁰ C · The solid dissolved to form a clear, colorless solution · The reaction mixture was then boiled for 2 hours at reflux, cooled to 25 ° G and Evaporated in vacuo to a yellow solid residue »This crude product was dissolved in 10 ml of ethanol, made basic with 0.1 N sodium hydrochloride, diluted with 500 ml of water and extracted four times with 50 ml of chloroform each time. The combined chloroform extracts were twice with 20 ml each Saturated sodium chloride solution withdrawn, filtered and evaporated in vacuo. The solid residue was suspended in 500 ml of ether, then filtered and hydrogen chloride gas in the pilate

- 72 -

009817/1888

initiated * The resulting precipitate was collected, dissolved in 3 ml of ethanol and reprecipitated by adding ether »

The solid was collected and dried to give in 30% yield (270 mg) 7 ~ 'B-dichloro-7-deoxylincomycin which was about 2 to 3 times the activity of lincomycin. In the Dünnschichtenchromatographie on silica gel (system methanol: chloroform _s Yolumenverhältnis 1: 6) is a spot having the Ef value 0.44 was obtained, he d Ef value for 7-chloro-7-deoxylincomycin is 0.52.

If lincomycin is replaced in this example by analogues of lincomycin according to formula II, where Z, R, R. ,, Ro, Rx of the group Ac have the above meaning, the corresponding 7-chloro «7- ^! ieoxyepilincomycin analogs

of the formula OH ₂ ,

σι

XXXIII

Most of the above-described compounds have 3 o / u it o in Jiipimer of the opposite configuration ₉ d _o ha

73 -

009817/1888

BATH ORIGINAL

the 7-epiform · .7 / becomes an inversion by substituting the 7-hydroxyl group caused by chlorine, so are the Epi-compounds which have an L-threo "configuration are converted into the derythro-configuration" In any case B- «erythro ~ and L-threο« forms can be obtained, depending after taking the normal Lincomycine (D-erythro) or the ] Spi-lincomycine (L-three) related «

Example 6;
7-chloro-7 "^{> (} 3eo3cylincomycin-hydrochloride

10 g (0.0226 mol) Idncoicycin-feydrochlorid, 200 ml carbon tetrachloride and 10 ml of _n Sogl warden stirring 4 hours refluxed. The reaction mixture was then cooled to 25 ° C. and filtered. The yellow solid was dried in vacuo and then dissolved in about 10 ml of boiling ethanol. Ethyl acetate was then added up to the beginning of Txubang, whereupon the solution was allowed to cool. In this way, crystalline 7- * chloro-7 ~ ⁽ ieox3rlincomycin-hyciiOchloria) was obtained in a yield of about 4 %.

0 0.98 1 7/1 888 '

OfiJGi _NA1

Claims (4)

Claims Process for the preparation of lincomycin derivatives of general formula in the R and Ac unreactive with thionyl chloride Remnants "mean, thereby germinated, that one Reduction of the formula AcIiK oei a temperature which allows the substitution of the? -hydrox; "l-.jruppe duich Jhlor err: c glicht,: z ± z Ihior ^ * Iohlorid un £ etsr, 009 817/1888 ■ 'BAD OBiQWAL 2. Method according to address 1, you du .: oh> kc: i.: I- drawn, datü R an alkylreat tüid Ac -,;.: · .J_cht with 'Jhi Chloride Reacting Acylrect ic-t. 3 · Method according to claim 1 'arr 2, dndirch; ■ -j; C · - ■ :::; draws that K is an alhvli'ect aj.i Ao =: ύ-Ι.ΐ: d-iirtelit, v.'otei iiZ a ctarlce acid · - i ^ t. 4. Method according to claim 1 - i, d- · ■ ;; ch ..; ■.-1: ·: rji- draws, dat s H a Α11ς; 1ΐ-βι.ΐ with never ·: *::.,?:;: ■ as 2C iCohlenstoxTato.Tien and Ac den Ac; ^r lrer; t ^ ir.rr ^l -cuti ^: tit-ai ten-L-2-PvrrolicincarlOnsäure dor? oru '- l :: (A) odor Uii represents, in which R, and R ^ alkylidenrecte:.: it not more than 20 carbon atoms, Oycloalkylidsnreste with 3 tis not more than S carbon atoms oi- = r ÄraIk; -Ii '· - width with not more than 12 carbon atoms Un- ?. R _x Jus be fabric or interpret blows. For THjC 'JhJCn.! OC-jrAIiY (LÜchican, VStA) 0 0 9 8 17/1