FI58502C - FOERFARANDE FOER FRAMSTAELLNING AV ALKYL-7-DEOXI-7- (ACORS) -ALFA-TIOLINKOSAMINIDER VILKA AER MELLANPRODUKTER VID FRAMSTAELLNING AV SUBSTITUERADE TIOANALOGER AV LINKOMYCIN - Google Patents

Description

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Petan * och registerstyrelsen · AmMom uth | deeh uti.sicrNtm puMorad 31.10.80 (32) (33) (31) Pyritty * »μΚμμ — Begirt prtocfm 29.06.71 I5.II.7I USA (US) 158075, 19901 * 6 ( 71) The Upjohn Company, 301 Henrietta Street, Kalamazoo, Michigan, USA (US) (72) Brian Bannister, Kalamazoo, Michigan, USA (US) (71 *) Berggren Oy Ab (51 *) for the preparation of alkyl-7-deoxy-7- (AcORS) -oi-thiolinkosamides - Preparation of alkyl-7-deoxy-7- (AcORS) - «- thiolinkosamines This invention relates to a process for the preparation of alkyl- (7S) -N-alkanoyl-2,3,4-tri-O-alkanoyl-7-deoxy-7-substituted thio-1-thio-L-threo-αD-galacto-octopyranosides, used as intermediates in the preparation of antibacterially active alkyl-6,7,8-trideoxy-7-substituted thio-6- (1-methyl-trans-N-propyl-L-pyrrolidin-2-ylcarbonylamino) -lt io-L-threo-α-D-galacto-octo-pyranoside, and whose intermediates have the formula cn3 y S-R-OAc

AcNH-- γΛ \ OAc / 13 / S-Alk OAc wherein Alk is lower alkyl, Ac is lower alkanoyl and R is a straight or branched alkyl group having 2 to 5 carbon atoms, an alkylthio-58502 alkyl group having 2 alkyl groups or 3 carbon atoms, or a cyclohexyl group.

The compounds of formula I are prepared by reacting alkyl-6,7-alkanalkylimido-6,7,8-tri-deoxy-1-thio-D-erythro-α-D-galacto-octo-pyranoside of formula

AcN ^^ D

^ - II

η / ~ \ \ 0Ac / \ j 1 / SAlk OAc where Alk is as defined above is heated with a cyclic sulfide of formula rRv where R is as defined above and 'S *' with an anhydrous acid having formula AcOH wherein Ac is lower alkanoyl. Thus, opening of the aziridine ring is provided to give an intermediate of formula I.

The acyl groups can then be removed by hydrazinolysis in a manner known per se (see U.S. Patent No. 3,179,565) to give the alkyl-7-deoxy-7- (HORS) -α-thiolincosamine of formula III.

^ CH

______x SR-OH

NH "-

2 III

©

Nj _ (/ S-Alk

OH

The compounds obtained from the intermediates prepared by the process of the invention (Formula III) are useful for the same purposes as the methyl α-thiolinososamide (methyl-6-amino-6,8-dideoxy-1-thio-) described in U.S. Patent No. 3,380,992. D-erythro-αD-galacto-octopyranoside, α-MTL) and methyl-6-amino-7-chloro-6,7,8-3,5502 trideoxy-1-thio-L-threo and D-erythro-αD- galactooctopyranosides (U.S. Patent Nos. 3,496,163 and 3,502,648), and the compound of formula III can be further acylated with trans-1-methyl-4-propyl-L-2-pyrrolidinecarboxylic acid to form 7-deoxy- 7 - (HORS) -lincomycin and with other L-2-pyrrolidinecarboxylic acids as described in these patents, or with N- (2-hydroxyethyl) -L-2-pyrrolidinecarboxylic acid to form compounds of formula CH , 3

_ / S-R-OH

(IV)

Ac- ^ NH-- γ \

VI ^ AlK

OH

wherein R and Alk are as defined above and Ac 1 is L-2-pyrrolidinecarboxyacyl or N-methyl, N-ethyl or N- (2-hydroxyethyl) -L-2-pyrrolidinecarboxyacyl, any or all of which may be substituted in the 4-position by lower alkyl or lower alkylidene.

It is known from U.S. Patent 3,544,551 that 7-SH analogs can be prepared by heating an aziridino compound of formula II wherein Ac is hydrogen with hydrogen sulfide. Until now, it has not been possible to replace S-hydrogen directly or indirectly. In addition, the compounds obtained via the intermediate of the invention are substantially much more active than the corresponding 7-SH compounds. For example, 7-deoxy-7 (S) - (methylthio) -lincomycin hydrochloride is several times more active in vitro than lincomycin, whereas 7-deoxy-7 - (S) -mercapto-lincomycin hydrochloride is less active than lincomycin.

It is also known from U.S. Patent 3,702,322 that 7-OR analogs can be prepared by reacting a compound of formula II with an alcohol in the presence of an acid. Attempts to make sulfur analogs by replacing alcohol with mercaptan have not been successful.

It has now been found that compounds of formula II undergo sulfidolysis when heated with a non-aromatic cyclic monosulphide in glacial acetic acid or other low alkanoic acid. The following formulas illustrate possible mechanisms in the reaction.

il 58502 ch3 ch3 ch3 jLa ------ S-CH, CHo0Ac + \ __ ^ OAc ^ 2 2

AcNH ----- AcNH ------

If the cyclic sulfide is asymmetric, as in the case of propylene sulfide, ch2ch2-ch3, two products are possible, e.g. CH CH,

! 3 CH 5 CH

I I 3 I 3 -1-S-CH-CH2OAc S-CH2-CH-OAc

AcNH-- AcNH-- | and

If the cyclic sulfide is oxacyclic or thiacyclic, an oxygen or other sulfur atom, X, is present in the side chain.

In cyclic sulfides, there is also the possibility that another mole of sulfide reacts as follows with ch3 ch3 --sC? Λ. --- S- (CH2) 3-S (CH2) 3-OAc (+) Sy y

AcNH-- AcNH--

OF

CH, t -5! -j-S- (CH2) 3OAc i

AcNH --f-- 5 58502 This reaction proceeds particularly well with trimethylene sulfide (titanium) to give approximately equal amounts of both products.

All of the above non-aromatic cyclic monosulfides provide the desired result simply by heating alkyl-N-alkanoyl-6,7-aziridino-6-deamino-7-deoxy-dc-thiolinincosamine in the presence of a suitable sulfide in the presence of glacial acetic acid or other anhydrous low alkanoic acid. .

Preferably, a solvent boiling at about 70 ° C is used. Excess sulfide is usually used for this purpose. If desired, solvents such as dioxane, carbon tetrachloride, benzene or toluene may be used, and preferably with a sulfide boiling above about 110 ° C. The pressure vessel can be advantageously used for lower boiling sulfides such as ethylene sulfide.

The ratios are not critical to the reaction, but they are critical to the yields. Thus, the best yields are obtained with about 3 to 7 ecyl alcohols of acid together with a substantial excess, at least twice the amount of sulfide. This is another advantage of using sulfide as a solvent. When using a sulfide, such as ethylene sulfide, which boils so low as to give a reaction mixture boiling below 70 ° C at reflux, an overpressure can be used. If it is such that the reaction mixture boils above about 110 ° C, controlled heating may be used. Otherwise, it is preferably heated to reflux and cooling temperature.

The reaction mixture can be further worked up by methods known per se in the art, such as countercurrent partitioning, chromatography and solvent extraction or crystallization.

The starting compounds exist in two epimeric forms: ch3 --CH3

Ac-N Ac-ϊζ ^ y- \ vr ^ \ yx

VjAlk V / SAIR

OAc OAc R-shape S-shape R and S refer to the 7-position because the 6-position is always in the R-shape. An inversion occurs in the reaction. When, for example, ethylene sulfide is reacted with methyl N-acetyl-2,3,4-tri-O-acetyl-6 (R), 7 (R) -aziridino-6-deamino-7-deoxy-N-thiolincosaminide, methyl N-acetyl-2,3,4-tri-O-acetyl-7-deoxy-7 (S) - (2-acetoxyethylthio) -A-thio-58852 lincosaminide.

The starting compounds of formula II are obtained by acylation of a compound of formula:

X

Ηϋ ^ 3 f)

\ * H i / sAlk OH

with a carboxyacyl acylating agent such as acetic anhydride or other low alkanoic anhydride or benzoyl chloride or the like carboxyacyl halide in a manner known per se. Because the amino and hydroxy groups are acylated at different rates, N-acyl and O-acyl may be the same or different.

Since these acyl groups are not present in the final product but are removed in the treatment, it is irrelevant what they are, as long as they are carboxyacyls. Suitable such carboxyacyls are low alkanoyls.

The starting compounds of formula VI can be prepared by dehydrohalogenating a compound of formula CH, 3 / \ ___ ✓ Halo NH2-- noj-o 'XoHySAlk

OH

which are known in the art. (U.S. Patent No. 3,502,648). Dehydrohalogenation is accomplished by heating a compound of formula VII in an inert solvent in the presence of an acid scavenger. A suitable way is to heat the reaction mixture of the starting material, anhydrous sodium carbonate and 7,550,502 dimethylformamide at reflux temperature for a short time, remove the solvent and crystallize from a suitable solvent, e.g. methanol (see U.S. Pat. No. 3,544,551).

Acylation of compounds of formula (III) from intermediates prepared by the process of the invention with L-2-pyrrolidinecarboxylic acid provides compounds of formula IV wherein Ac is L-2-pyrrolidinecarboxylic acid acyl. When Alk is methyl, R is ethylene and L-2-pyrrolidinecarboxylic acid is trans-1-methyl-4-propyl-L-2-pyrrolidinecarboxylic acid and the configuration is (S), the compound is 7-deoxy-7 (S ) - '(2-hydroxyethylthio) -lincomycin, which has several times greater antibacterial activity than lincomycin. It and its analogues can be used for the same purposes and in the same way as lincomycin. Lower members have better gram-negative activity. Table 1 shows the M.I.C. (minimum inhibitory concentration) of lincomycin and its 7 (S) -7-substituted thioanalogs for the three bacteria that can still kill or inhibit the growth of the bacterium under certain conditions. In addition to the final products obtained via the intermediates according to the invention, other lincomycin analogues are also shown in the experimental results.

8 58502

M.I.C. values (μg / ml) of (7S) -7-substituted thio analogues of lincomycin compared to lincomycin

Streptococcus Streptococcus Staphylococcus pneumoniae pyogenes aureus

Lincomycin 0.31 0.31 0.31

Lincomycin Eg No. thio analogues (intermediate, R-group from which the final product (obtained in formulas) III and IV)

Et 1 0.04 0.08 0.04 n-Pr 2 0.01 0.04 0.02 i-Pr 3 0.02 0.02 0.02 cyclohexyl 5 0.125 0.125 0.125 n-Bu 4 0.02 0.125 0.02

The invention is described in more detail below by means of examples in which parts are given by weight, except in the case of solvent ratios or unless otherwise stated and the c.g.s. system is used unless otherwise stated.

Example 1

Methyl N-acetyl-2,3,4-tri-O-acetyl-7 (S) - (2-acetoxyethylthio) -7-deoxy-α-thiolincosaminide 9 58502 CH, CH, 3 3

AcN ^ --SCH2CH2OAc _ AcNH-- w · yi OAc

A mixture of 5.0 g (1 molar equivalent) of methyl N-acetyl-2,3,4-tri-O-acetyl-6,7-aziridino-6-deamino-7-deoxy-α-thio-linco- amide, 50 cm -1 of ethylene sulfide (titanium) and 5.25 g (7 molar equivalents) of glacial acetic acid are heated in a test tube for 20 hours on a steam bath. The volatiles are removed from the reaction solution by distillation at 100 ° C, the residue is dissolved in methylene chloride and mixed with an excess of saturated aqueous sodium bicarbonate solution. Washing the organic layer with water, drying over anhydrous sodium sulfate and removing the solvent on a rotary evaporator at 40 ° C / 7 mm Hg gives a thick yellow syrup which, according to thin layer chromatography (SiO 2 gel, acetone and "Skellysolve B" in a ratio of 1: 1 (" Skelly-solve B "is essentially n-hexane (b.p. 60-68 ° C, manufactured by Skelly Oil Co.) does not contain any starting material as well as a new main zone with a lower Rf value (0.50 compared to 0.69 : reads).

The above syrup, which contained a substantial amount of polymeric material derived from ethylene sulfide, is chromatographed to remove the polymeric material. Use a column containing 1200 g of silica (5.8 x 89 cm) and a solvent system with ethyl acetate and "Skellysolve B" in a ratio of 2: 1. After one liter of pre-run, 50 ml fractions are collected until all polymeric material has been removed by thin layer chromatography.

The column is then eluted with ethyl acetate. Fractions 81-170 are combined and the solvent is removed by distillation on a rotary evaporator at i40 ° C / 7 mm Hg to give 5.81 g of crude product.

Countercurrent partitioning of the crude product in system 1 ethanol: 1 water: 1 ethyl acetate: 3 cyclohexane gives methyl N-acetyl-2,3,2-tri-O-acetyl-7 (S) - (2-acetoxyethylthio) -7-deoxy-α- thiolinkosaminide with a K value of 0,53 · It is obtained as colorless needles from ethyl acetate "Skellysolvolv B" and has the following properties: 10 58502

Sp. 206-7 ° / d? + 180 (c, 0.79, CHCl 3)

Analysis: Calculated for C 48.17, H 6.35, N 2.68, S 12.25.

Found: C 48.12, H 6.37, N 2.58, S 11.95-

Example 2

Part A Methyl N-acetyl-2,3,4-tri-O-acetyl-7 (S) - (3-acetoxy-propylthio) -7-deoxy-α-thiolincosamide

By proceeding as in Example 1, substituting trimethylene sulfide for ethylene sulfide, methyl N-acetyl-2,3,4-tri-O-acetyl-7 (S) - (3-acetoxypropylthio) -7 'deoxy-α-thiolincosaminide (K = 1.0 , 1 ethanol: 1 water: 1 ethyl acetate: 2 cyclohexane) as colorless needles ethyl acetate: "Skellysolvesta B" and with the following characteristics:

Sp. 172.5-174 ° / £ 7d + 178 ° (c, 0.94, CHCl 3)

Analysis: Calculated for C22H35ONS2: C 49.15, H 6.56, N 2.61, S 11.93

Found: C 49.31, H 6.58, N 2.68, S 11.83.

Part B Methyl N-acetyl-2,3,4-tri-O-acetyl-7 (S) - [3- (3-acetoxy-propylthio) propylthio] -7-deoxy-α-thiolincosaminide Methyl N- acetyl-2,3,4-tri-O-acetyl-7 (S) - [3- (3-acetoxypropylthio) propylthio] -7-deoxy-α-thiolincosaminide (K = 2,3, same solvent system) as colorless needles has the following characteristics:

Sp. 117-8 ° / £ 7d + 149 ° (c, 0.997, CHCl 3)

Analysis: Calculated for C 9 H 18 N 2 O 3 S: C 49.08, H 6.76, N 2.29, S 15.72.

Found: C 49.38, H 6.14, N 2.35, S 15.91-

Example 3

Methyl 7 (S) - (2-acetoxy-1-methylethylthio) -N-acetyl-2,3,4-tri-O-acetyl-7-deoxy-α-thiolinincosamide and methyl 7 (S) - (2 -acetoxy-2-methylethylthio) -N-acetyl-2,3,4-tri-O-acetyl-7-deoxy-α-thiolincosamide

By proceeding as in Example 1, substituting propylene sulfide (1,2-epithiopropane) for ethylene sulfide, a mixture of 40 parts of methyl 7 (S) - (2-acetoxy-1-methylethylthio) -N-acetyl-2,3,4- tri-O-acetyl-7-deoxy-α-thiolincosaminide and 60 parts of methyl 11,5 8502 7 (S) - (2-acetoxy-2-methylethylthio) -N-acetyl-2,3,4-tri-O- acetyl 7-deoxy-α-thiolincosaminide (K = 0.83, 1 ethanol: 1 water: 1 ethyl acetate: 3 cyclohexane) as small colorless needles Ethyl acetate: From "Skellysolve B" and the mixture of isomers had the following properties:

Sp. 198-199 ° C.

[α] D + 170 ° (c, 0.94, CHCl 3)

Analysis: Calculated for C 22 H 35 O 10 NS 2: C 49.15, H 6.56, N 2.61, S 11.93.

Found: C 49.41, H 6.47, N 2.34, S 11.53.

Example 4

Methyl-7 (S) - (4-asetoksibutyylitio) -7-deoxy-a-tiolinkosaminidi

Following the procedure as in Example 1, substituting tetrahydrothiophene (tetramethylene sulfide) for ethylene sulfide at 100 ° C for 20 hours gives methyl N-acetyl-2,3,4-tri-O-acetyl-7 (S) - (4-acetoxybutylthio) -7- deoxy-α-thiolincosaminide (K = 1.32, 1 ethanol: water: 1 ethyl acetate: 2 cyclohexane) as fine needles ethyl acetate: "Skellysolve B" and with the following properties:

Sp. 149-150 ° C.

/ §7d + 171 ° (c, 0.88, CHCl 3)

Analysis: Calculated for C 23 H 37 O 10 NS 2: C 50.07, H 6.76, N 2.54, S 11.62.

Found: C 49.97, H 6.86, N 2.50, S 11.35.

Example 5

Methyl N-acetyl-2,3,4-tri-O-acetyl-7 (S) - (2-acetoxycyclohexylthio) -7-deoxy-α-thiolinincosamide

By proceeding as in Example 1, substituting cyclohexene sulfide (7-thiabicyclo- / 5.1.0-heptane) for ethylene sulfide and heating at 100 ° C for 16 hours, methyl N-acetyl-2,3,4-tri-O-acetyl-7 (S ) - (2-asetoksisykloheksyylitio) -7-deoxy-a-tiolinkos-aminidia. Countercurrent partitioning in a solvent system 1 ethanol: 1 water: 0.5 ethyl acetate: 3 cyclohexane gives this product with a K value of 0.80. It is obtained as colorless needles from ethyl acetate and has the following properties:

Sp. 205-6 ° C.

/ $ 7d + 153 ° (c, 0.64, CHCl 3)

Analysis: Calculated for C 25 H 39 O 10 N 2 O 2: C 51.97, H 6.80, N 2.43, S 11.10,

Molecular weight 577.70.

12 58502

Found: C 51.82, H 6.87, N 2.29, S 11.12,

Molecular weight (mass spectrum, M +) 577 ·

Example 6

Methyl N-acetyl-2,3,4-tri-O-acetyl-7 (S) - (5-acetoxypentylthio) -7-deoxy-α-thiolincosamide

By a procedure as in Example 1, pentamethylene sulfide (tetrahydrothiopyran) is reacted to methyl N-acetyl-2,3,4-tri-O-acetyl-7 (S) - (5-acetoxypentylthio) -7-deoxy-α-thiolinkosamide. with a K value of 1-9 * 1 1 ethanol: 1 water: 1 ethyl acetate: 2 in cyclohexane and having the following properties crystallized from ethyl acetate: "Skel ly solve B":

Sp. 158-9 ° C. (needles) / £ 7d + 169 ° (c, 0.60, chci3)

Analysis: Calculated for C 24 H 39 O 10 NS 2: C 50.95, H 6.95, N 2.48, S 11.34.

Found: C 50.88, H 6.98, N 2.41, S 11.22.

Example 7 (Preparation of the final product) 7-Deoxy-7 (S) - (2-hydroxyethylthio) -1-endomycin hydrochloride / Methyl-6,7,8-trideoxy-7- (2-hydroxyethylthio) -6-trans- (1-methyl- 4-propyl-L-2-pyrrolidiinikarboksiamino) -l-thio-L-threo-a-D-galaktopyranosidivetykloridi? m Me CH3 I. HCl M CH, --SCH2CH2-OH \ -5 h2n-- h - / - sch2ch2-oh HO A °, CONH-- l \) H / H0 / ° vv; _ / SMe Me, 1 / \ l III | \ OH / OH / N \ V. y SMe

Cr

XVIII \ j j / CO · 0 »CQ-0’isoBu XIX

To a suspension of 930 mg of trans-propyl hygric acid hydrochloride in 50 ml of anhydrous acetonitrile is added 995 mg of triethylamine. After the solid has dissolved, the solution is cooled to -5 ° C in an ice-methanol bath, which causes the separation of triethylammonium chloride. To this solution is added 610 mg of isobutyl chloroformate so that the temperature does not exceed -3 ° C, and the reaction mixture is stirred at a temperature between -3 ° C and -5 ° C for 20 minutes. 700 mg of methyl 7-deoxy-7 (S) - (2-hydroxy-ethylthio) -α-thio-lincosaminide in 5 g of methanol and 5 g of water are then added to the mixed anhydride, whereby the precipitate of triethylammonium chloride dissolves immediately. After 2 hours, thin layer chromatography (SiO 2 gel, 1 MeOH / 10 CHCl 3) shows that the amino sugar has faded and a new band has developed with a higher Rf value (0.30). The volatile solvent is removed on a rotary evaporator at 40 ° C / 7 mm Hg and the syrupy residue is dissolved in a mixture of water and methylene chloride and the pH of the aqueous layer is adjusted to 10 with $ 50 aqueous sodium hydroxide solution. The aqueous solution is separated and extracted twice with methylene chloride, and the combined extracts are dried over anhydrous sodium sulfate. The remaining basic aqueous layer is discarded.

Removal of the solvent from the methylene chloride extract on a rotary evaporator at 40 ° C / 7 mm Hg gives a pale yellow syrup which is chromatographed on silica (480 g, column dimensions 3.8 x 95 cm, lag volume 850 cm -1) in a system of 1 methanol: 1 chloroform. 250 3 ... 3.

After a pre-run of 1 cm, 50 cm fractions are collected and elution is followed by thin-layer chromatography on silica in the same system.

Fractions Nos. 25 to 44 are combined to give, after removal of the solvent, 610 mg of a colorless glass which is reacted to give the hydrochloride by dissolving it in water to which a dilute aqueous solution of hydrochloric acid (N) is added until the pH of the resulting solution is about 3 *. after yielding 7-deoxy-7 (S) - (2-hydroxyethylthio) -lincomycin hydrochloride (C-3) as a colorless solid having the following properties: [α] D + 114 ° (c, 0.91, H 2 O)

Analysis: Calculated for ^ 20 ^ 38 ^ 6 ^ 2 ^ 2: C 47.74, H 7.81, N 5.57, Cl 7.05, S 12.75,

Molecular Weight with Free Base 466.65 ·

Found (corrected for $ 6.75 for H 2 O): C 48.05, H 7.70, N 5.10, Cl 6.96, S 12.50,

Molecular weight (mass spectrum, M + with free base) 466. Biological activity: about 8 times lincomycin in a test tube. This compound is about 8 times more active than lincomycin and has a higher gram-negative activity in the living organism and is less toxic than 7-deoxy-7 (S) -chlorlininkomycin hydrochloride.

Claims (1)

Process for the preparation of alkyl- (7S) -N-alkanoyl-2,3,4-tri-O-alkanoyl-7-deoxy-7-substituted thio-1-thio-L-threo-αD-galactooctopyranosides which used as intermediates in the preparation of antibacterially active alkyl-6,7,8-trideoxy-7-substituted thio-6- (1-methyl-trans-4-propyl-L-pyrrolidin-2-ylcarbonylamino) -1-thio- L-threo-αD-galacto-octopyranoside, and whose intermediates have the formula CH, ^ --- ^ y SR-OAc AcNH-- (I) Acor N v OAc / \ j_V S-Alk OAc where Alk is low alkyl, Ac is lower alkanoyl and R is a straight or branched alkyl group having 2 to 5 carbon atoms, an alkylthioalkyl group having 2 or 3 carbon atoms in the alkyl group, or a cyclohexyl group, characterized in that the alkyl-6,7-alkanoylimido -6,7,8-tri-deoxy-1-thio-D-erythro-αD-galacto-octopyranoside of the formula AcN 3 (II) Ac ° / ° \ | V \ | 0Ac | / sAlk OAc \ where Alk is as defined above is heated with a cyclic sulfide of formula ^ R »where R is as defined above and V with an anhydrous acid of formula AcOH where Ac , is lower alkanoyl.