DK169681B1 - Prepn. of pristinamycin II B s-oxide derivs. - by oxidising corresp. thio-substd. deriv. with potassium peroxy:mono:sulphate - Google Patents

Abstract

Prepn. of a pristinamycin IIB S-oxide deriv. of formula (I), including isomeric forms or mixts. and acid addn. salts, comprises oxidising a pristinamycin IIB deriv. of formula (II), or salt or protected deriv., with potassium peroxymonosulphate, and opt. sepg. isomers, deprotecting and/or salifying: where R = (i) 4- to 7-membered N-contg. heterocycle opt. contg. further N, O and/or S (as sulphoxide or sulphone) heteroatom(s), and opt. substd. by alkyl; or (ii) 2-4C alkyl, substd. by 1 or 2 of phenyl, cycloalkylamino or N-alkyl-N-cycloalkylamino with 3-6 ring C, alkylamino, dialkylamino or dialkylcarbamoyloxy (these latter two opt. having the alkyl gps. joined to form a gp. as defined for (i) above with opt. 1 futher heteroatom), or substd. by 1 or more gps. as for (i) contg. 1 or 2 further heteroatoms as above and bonded via C, at least one of the substits. on the alkyl being an N-contg. substit. capable of forming salts; or (iii) an ((S)-1-methyl-2 -pyrrolidinyl) methyl gp.; the alkyl gps. being 1-10C opt. branched alkyl. In (II), any S atoms in heterocyclic substits. in R may be in S, SO or SO2 form. (I) and (II) are known from EP-191662.

Description

in DK 169681 B1

The present invention relates to a particular process for the preparation of pristinamycin IIB-S-oxide derivatives of the general formula I

0 (I) cVcx CH, R-SO 2 S / ° 10 wherein R means a) a nitrogen-containing heterocyclyl group of 4-7 ring members optionally containing one or more other heteroatoms selected from nitrogen, oxygen and sulfur in the form of sulfoxide or or b) an alkyl chain of 2-4 carbon atoms substituted by one or two groups selected from phenyl, cycloalkylamino or N-alkyl-N-cycloalkylamino containing 3-6 ring members; 20 alkylamino, dialkylamino and dialkylcarbamoyloxy (the alkyl moieties of the latter two groups optionally together with the nitrogen atom to which they are attached may form a saturated or unsaturated 4-7 ring heterocyclic group optionally containing another heteroatom selected from the nitrogen group. (C) an alkyl chain of 2 to 4 carbon atoms substituted by one or more nitrogen-containing heterocyclic groups, such as sulfoxide or sulfone, optionally substituted by an alkyl group) or are with 4-7 ring members optionally containing one or two other 30 heteroatoms selected from nitrogen, oxygen and sulfur in the form of sulfoxide or sulfone and optionally substituted with an alkyl group, said heterocyclic groups being bonded to the chain which carrying them, via a carbon atom, or d) a [1-methyl-2 (S) -pyrrolidinyl] methyl group, at least one of the substituents carried by said alkyl chain being a nitrogen-containing substituent which may form salts and the above-mentioned alkyl groups are straight or branched and contain 1-10 carbon atoms, unless otherwise stated, in the form of their isomers or their mixtures - or their addition salts with acids, which process is peculiar by the characterizing part of claim 1.

Where R in the general formula I means a heterocyclyl group, this group may e.g. be selected from 3-azetidinyl, 3-pyrrolidinyl, 3- or 4-piperidyl or 3- or 4-azepinyl.

When R is a heterocyclylalkyl group, the heterocyclyl group may e.g. may be selected from the above groups or 2-azetidinyl, 2-pyrrolidinyl, 2-piperidyl, 2-azepinyl, piperazinyl, 4-alkylpiperazinyl, quinolyl, isoquinolyl or imidazolyl.

When R contains a dialkylamino or dialkyl carbamoyloxy group whose alkyl moieties together with the nitrogen atom to which they are attached form a heterocyclic group, e.g. selected from 1-azetidinyl, 1-pyrrolidinyl, piperidino, 1-azepinyl, morpholino, thiomorpholino in the form of sulfoxide or sulfone, 1-piperazinyl, 4-alkyl-1-piperazinyl, N-alkyl-1-homopiperazinyl and 1-imide-azolyl.

EP Patent Application No. 191,662 discloses compounds similar to the compounds of formula I.

According to the EP patent application, the compounds of formula I are prepared by oxidation of a pristinamycin lig derivative or a salt or a protected derivative thereof with the general formula II:

DK 169681 B1 3

As oxidizing agents Can be used organic peracids or inorganic peracids, e.g. persulphuric.

It has now been found that the compounds of the general formula I can be prepared by oxidizing a pristinamycin lig derivative of the general formula C by means of the potassium monopersulfate in the form of the triple salt [2KHSO5 · KHSO4 · K2SO4]. R 5 in which R is as defined above, except that when R contains a sulfur-containing heterocyclic group, the sulfur atom may also be in the form of a sulfide or a salt or protected derivative thereof, and optionally then separate the resulting isomers.

20 Potassium monopersulfate in the form of the triple salt [2KHSO5 · KHSO4 · K2SO4] is sold under the name "Oxone".

Taking into account the benefits obtained with "Oxone", namely ease of use and increased safety over compressed sulfuric acid, it may be obvious to one skilled in the art to use this oxidizing agent.

However, given the publications in which the use of this oxidant is mentioned, it would be expected that it would not be useful for the oxidation of compounds of formula II.

Namely, pristinamycins are complex molecules that contain numerous oxidizable functions (double bonds, conjugated double bonds, hydroxy groups). R.J. Kennedy, J. Org. Chem. 25, 1901 (1960) (ref. I) -, shows that numerous functions are oxidized by "Oxone" (sulfide, double bonds, alcohols, ketones, amines). R. Bloch et al., J.

Org. Chem. 50, 1544 (1985) (ref. 2), shows that "Oxone" gives rise to a considerable amount of epoxide in DK 169681 B1 4 when the molecule contains an unsaturation: "We now report that potassium hydrogen persulfate alone is capable of to epoxidize water-soluble or insoluble alkenes "with good to excellent yields, opening a new, effective and simple path for the synthesis of epoxides."

Various examples listed in Table I of this publication show, among other things, epoxidation of conjugated systems of double bonds.

According to the prior art, oxidations by means of "Oxone" lead in particular to sulfones (see ref. 1 and 2 above), while the reaction according to the present invention largely leads to sulfoxide.

It was also conceivable that amino group-containing chains substituted on the sulfur atom would degrade during oxidation, either by elimination of amine or by oxidation of nitrogen (see ref. 1).

Thus, it is surprising that "Oxone" can lead to selective oxidation of the sulfur function in pristinamycins to a sulfur oxide function, but it has also been found that this oxidation preferably leads to the formation of isomer A2 and allows products with better purity. Thus, it has become possible to crystallize the products directly after preparation and avoid the previously necessary successive chromatographs. The above is explained in more detail later in the description.

The reaction is carried out in water or in a mixture of water and alcohol (eg water / methanol) or water and chlorinated solvent (eg water / dichloromethane) at 30 temperatures between -60 and + 25 ° C.

When the pristinamycin lig derivative of general formula II is used in the form of a salt thereof, salts formed with organic or inorganic acids, preferably with trifluoroacetic acid, tartaric acid, acetic acid, benzoic acid, hydrochloric acid or sulfuric acid, or with potassium hydrogen sulfate are used.

When R contains an alkylamino or cycloalkylamino nosubstituent, it is also possible to use a protected derivative of the compound of general formula II, which may be protected by any amine protecting group whose introduction and removal does not affect the residue of the molecule. Advantageously, there is used a trifluoroacetyl group which, after reaction, can be removed by treatment with an alkali metal hydrogen carbonate (sodium or potassium hydrogen carbonate) in aqueous solution.

The potassium monopersulfate is prepared by the method described in US Patent No. 2,802,722.

The compounds of general formula II can be prepared by the method described in EP Patent Application No. 191,662. It is not absolutely necessary to isolate the pristamicamycin β-derivative of the general formula II to use it in the process of the invention.

The compounds of general formula I obtained by the present process can be purified by known methods, e.g. by crystallization, chromatography or successive extractions in acidic or basic medium. In view of the sensitivity of the basic medium to the synergistins, basic medium is understood to mean a medium which is just sufficiently basic to release the compound from its addition salt with an acid, ie. a medium whose pH does not exceed 8.

The isomers of the compounds of general formula I

can be separated by any known method. Advantageously, chromatography or high-pressure liquid chromatography is used.

The compounds of general formula I obtained by the method of the invention exhibit antibacterial activity against gram-positive bacteria (staphylococci, streptococci, pneumococci, enterococci) and gram-negative bacteria (haemophilus, gonococci, meningococci). In addition, they exhibit the advantage that they can be dissolved in water, generally in the form of salts, at useful therapeutic doses 35 and exhibit a synergistic antibacterial effect with pristi-namycin 1, virginiamycin S, or soluble synergistins in waters such as described in EP Patent Application No. 191,662, page 16.

Furthermore, the compounds of general formula I prepared by the process of the invention may

5 may be useful in the preparation of pristinamycin IIB sulphonic derivatives of the general formula III

Wherein R has the same meaning as in formula I, which derivatives are active as antimicrobial agents and also exhibit a synergistic effect together with group I pristinamycins.

The pristinamycin IIB sulfone derivatives of general formula III are prepared by oxidation of a pristinamycin lig derivative of general formula I under the conditions described in EP patent application No. 191,662.

In particular, in the process of the invention, compounds of general formula I are prepared wherein R is an alkyl chain of 2-4 carbon atoms substituted by one or two groups selected from phenyl, cycloalkylamino or N-alkyl-N-cycloalkylamino containing 5 or 6 ring link, alkylamino of 1-4 carbon atoms, dialkylamino (whose alkyl moieties contain 1-3 carbon atoms or together with the nitrogen atom to which they are attached form a saturated heterocyclic group of 5 or 6 ring members), or R means a 5- or 6- a nitrogen-containing heterocyclic group optionally substituted with an alkyl group of 1-4 carbon atoms, at least one of the substituents carried by the above alkyl chain being a nitrogen-containing substituent capable of forming salts and at least one of the groups supported by this chain are placed in 1-position or 2-position.

The compounds of the general formula I thus prepared can be converted into acid addition salts thereof.

In addition, as pharmaceutically acceptable salts, addition salts may be mentioned with inorganic acids such as hydrochlorides, hydrobromides, sulfates, nitrates or phosphates, or with organic acids such as acetates, propionates, succinates, maleate, fumarates, methanesulfonates, p-toluenesulfonates, isethionates , citrates or tartrates or substitution derivatives of these compounds.

The invention is illustrated by the following examples. The NMR spectra of the compounds illustrated in these examples exhibit general characteristics common to all the compounds of general formula I and particular characteristics specific to each of the compounds depending on the substituents. In the examples, only the special characteristics that result from variable groups are listed.

For the compounds of general formula I, the protons 20 are denoted by the numbering given in the following formula: 0 V-'T'n Ry „8 10> 53 lis o 25 32 0 0 j ^ 6 s CH 2 20 ° "30

Unless otherwise stated, all spectra are recorded at 250 MHz in deuterochloroform. The chemical shifts are expressed in ppm relative to the tetramrethylsilane signal.

The abbreviations used below have the following meanings: DK 169681 B1 8 s = singlet d = doublet t = triplet mt = multiplet 5 ~ m = solid dd = doublet of duplicates dt = duplicate of triplets ddd = duplicate of duplicates of duplicates dddd = duplicate of duplicates of duplicates of duplicates.

10

The various isomers are arbitrarily classified according to the chemical shifts observed in the NMR spectra.

As isomer A 1 and isomer A 2 of the compounds of the general formula I are isomers having the following characteristics: 1.7 (s, -CH 3 in 33); ca. 3.8 (s, 2 CH 2 in 17); <5 (d, -H27) isomer A2 or> 5 (d, -H27) isomer A2; ca. 5.50 (d wide, -H13); ca. 6.20 (d, -Ηχχ); ca. 6.6 (2H NH in 8); > 8 (s, 20-H2O).

Isomers and isomers B2 of compounds of general formula I are termed isomers which exhibit the following characteristics; ca. 1.5 (s, -CH 3 in 33); ca. 3.7 and 3.9 (2d, TCH2 in 17); 25 approx. 4.8 (mt, -H13); <5 (d, -H27) isomer B2 or> 5 (d, -H27) isomer Blf · approx. 5.70 (boundary AB, -H 2 and -H 10); ca.

7.7 C 2 NH 8); ca. 7.8 (s, -H 2 O).

As isomer A of the compound of general formula II, the isomer is shown which exhibits NMR characteristics identical to those listed above for isomers A 1 and A 2 of the compounds of general formula I, with H in the 27 position. characterized by 4.7 (d, J <1 Hz).

As isomer B of the compound of the general formula II, the isomer having NMR characteristics identical to those listed above for the isomers B 1 and B2 of the compounds of the general formula I is denoted as H in 27-style DK 169681 B1 9 ling is characterized by 4.6 (d, J> 2.5 Hz).

Example 1

To 13 g of 26- (2-diethylaminoethyl) -thiopristinamycin 5 lig (isomer A) suspended in 170 ml of distilled water are added over 15 minutes at 0 ° C to 40 ml of an aqueous solution of 8.1 g of Oxone . The resulting mixture is stirred for 30 minutes at 0 ° C, then 1.3 g of "NORIT® SX ULTRA" - black and a small amount of sodium thiosulfate are added. After 30 minutes-10 hours stirring at 20 ° C, the suspension is filtered through Ce-lite® and then rinsed with 50 ml of distilled water. The solution is adjusted to a pH of 7 by the addition of solid sodium bicarbonate and then washed with 3 times 100 ml of dichloromethane. The organic phases are combined, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C to give 9.9 g of a light beige solid containing 85% 26- ( 2-diethylaminoethyl) sulfinylpristinamycin lig (isomeric A2) / 10% isomer A 2 and 5% 26- (2-diethylaminomethyl) sulfonylpristinamycin II3.

The crystalline form of 26- (2-diethylaminoethyl) sulfinylpristinamycin II3 can be obtained as follows.

5 g of the solid prepared above are taken up in 13 ml of acetonitrile. After dissolving in the heat, 25 ml of ether are added and crystallization is started by scraping.

The crystals are filtered off, washed with ether and then dried under reduced pressure (270 Pa) at 20 ° C. In this way, 3.9 g of 26- (2-diethylaminomethyl) sulfinylpristiamycin is obtained (isomeric A 2: 85%, isomeric A 2: 15%) in the form of 30 white crystals melting near 116 ° C.

NMR Spectrum (isomer A2): 1.03 (t, -N (CH2 CH3) 2) 1.75 (S, -CH3 in 33) 2.05 and 2.55 (2mt, 45-2.70 (mt, -N (CH 2 CH 3) 2) 2. 169 - 3.10 (mt, -SCH 2 -CH 2) o 2.75 (mt, H i 4) 2.92 - 3 Δ, 3.22 (mt, H i 26) 3.82 (S,> CH 2 i 17) 4.81 (d, H i 27) 5.5 (d, H i 13) 6.19 (d, H in 11) 6.50 (dd, TN TNH in 8) 6.58 (dd, H in 5) 8.12 (s, H in 20) NMR Spectrum (isomer A J: 1.04 (t, -N (CH2 CH3) 2) 1.69 (S, -CH3 in 33) 2-2.3 (mt, ^: CH2 in 25) 2.60 (mt, ^ N-CH2 -CH3) 2.7-2.95 (mt, -S (O) -CH2-CH2-NiC,) 2.7 (mt, H in 4) 2.86 and 3.04 (2dd, 15) 3.28 (mt, H i 26) 3.78 (AB system, ^ CH 2 i 17) 5.25 (d, H i 27) 5.4 (d, H i 13) 6.15 ( d, H in 11) 6.60 (dd, H in 5) 6.83 (dd, / NH in 8) 8.08 (S, H in 20) 30- (2-Diethylaminoethyl) -thiopristinamycin lig can be prepared as described in U.S. Pat. U.S. Patent No. 4,590,004.

Example 2

To 3 g of 26- (2-diethylaminoethyl) -thiopristinamycin 35 IIB (isomer A) suspended in a mixture of 30 ml of distilled water and 6 ml of ethanol are added over 5 minutes at -5 ° C.

DK 169681 B1 11 10 ml of an aqueous solution of 1.9 g of "Oxone". After 10 minutes of stirring, the reaction mixture is washed with 2 times 20 ml of dichloromethane. The aqueous phase is adjusted to a pH of 7 by the addition of solid sodium bicarbonate 5 and then extracted with 4 times 50 ml of dichloromethane. The organic phases are combined, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 40 ° C. In this way, 2.4 g of a white solid containing 26- (2-diethylaminoethyl) sulfinylpristamycin IIB (isomer A2: 88%, isomer A ^: 6%) and 6% 26- ( 2- (diethylaminoethyl) sulfonylpristinamycin Hg. The product has the same NMR characteristics as the product of Example 1.

Example 3

To 3.2 g of 26- [1-diethylamino-2 (R) -propyl] -thiopristiamycin IIB (isomer A: 87%, isomer B: 13%) dissolved in 48 ml of methanol is slowly added at -30 ° C. , 65 g of Oxone "dissolved in 32 ml of distilled water. After 30 minutes at -30 ° C and then 20 after 15 minutes at -40 ° C, 30 ml of distilled water are added and then 3 g of" NORIT® SX ULTRA "variety The mixture is stirred for 30 minutes at 20 ° C, filtered over "Celite®", then rinsed with 50 ml of distilled water. The aqueous phase is washed with twice 100 ml of ethyl acetate and then 50 ml of ethyl ether are adjusted to a pH value. of 7 by the addition of solid sodium bicarbonate and extracted with 250 ml of dichloromethane twice, the organic phases are washed with 50 ml of distilled water, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C. In this way, 2.4 g of a very pale yellow meringue-like substance is obtained which is stirred in 40 ml of a mixture of pentane and diethyl ether in volume ratio After filtering and drying under reduced pressure (270 Pa) at 20 ° C, 2.2 g of 26- [1-diethylamino-2 (R) -propyl] -sulfinylpristamycin IIB (isomer A: 80) is obtained. %) in the form of an off-white solid which melts near 140 ° C.

DK 169681 B1 12 NMR Spectrum: 1.02 (t, -N (CH 2 -CH 3) 2)

S H

5 - 1,34 (d,) O - CH 2 - N (C 2 H 5) 2

Cf3 1.72 (S, - CH3 in 33) 2.01 and 2.55 (2mt, 1 ^ CH2 in 25) 2.45 - 2.70 (mt, 1H of -CH2-N (CH2-CH3) 2 and -N (CH2 - "CH3) 2) 2.90 (mt, 1H of -CH2-N (CH2CH3) 2) 2.76 (mt, H in 4) 2.88 and 3.08 (2dd, i 15) 3 (mt, -S-CH-)} 1 "o ch3 3.73 (mt, H i 26) 3.80 (s, ^: ch2 i 17) 4.92 (s wide, Η i 27) 5.42 (d, H i 13) 6.15 (d, H i 11) 6.55 (dd, H i 5) 6.70 (dd, H NH i 8) 8.06 (s, H i 20) 26- [1-Diethylamino-2 (R) -propyl] -thiopristinamycin may be prepared as follows.

To 10.5 g of pristinamycin IIA suspended in 200 ml of methanol is added at -30 ° C under nitrogen 3.2 ml of 1-diethylamino-2 (R) -propanthiol. After stirring for 18 hours at -30 ° C, 2 ml of methyl acrylate is added and stirred for 1 hour. Then 200 ml of distilled water, potassium hydrogen sulfate are added to a pH of 4 at -10 ° C and then 10 g of 35 "NORIT® SX ULTRA" variety. After stirring for 30 minutes at 0 ° C, the mixture is filtered through "Celite®" and then washed with 150 ml of distilled water. The aqueous phase is washed with 100 ml of ethyl acetate and then 100 ml of ethyl ether, then adjusted to a pH of 7 by the addition of solid sodium bicarbonate. After extraction with 200 ml of dichloromethane, the organic phase is washed with 100 ml of water, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C. The resulting solid is evaporated in 100 ml of ethyl ether, filtered on a glass frit and then dried under reduced pressure (270 Pa) at 20 ° C. This gives 3.36 g of 26- [1-diethylamino-10 2 (R) -propyl] -thiopristinamycin IIB (isomer A: 85%, isomer B: 15%) in the form of a light beige solid which melts near 130 eC.

NMR Spectrum (isomer A): 1.03 (t, -N (CH 2 CH 3) 2)

15 \ -N

1.32 (d,) T + (CH2CH3) 2 CH3 1.72 (S, -CH3 in 33) 1.88 and 2.10 (2mt, I> CH2 in 25) 2.55 (mt, -N (CH2CH3) 2) 2.45 and 2.60 (2mt, -CH2-N (CH2CH3) 2) 2.78 (mt, H in 4) 2.92 and 3.10 (2dd, 1 ^ CH2 in 15) 3 (mt, -S-CH-) CH3 3.52 (rnt, H in 26) 3.82 (S, CH2 in 17) 4.79 (s wide, H in 27) 5.49 (d, H i 13) 6.13 (d, H i 11) 6.32 (m, NH i 8) 6.52 (dd, H i 5) 8.13 (S, H i 20) DK 169681 B1 14 (isomer B): 1 (t , -n (CH 2 CH 3) 2)

\ _N

1.50 (s, -CH3 in 33) 2.05 and 2.5 (2mt, ^ CH2 in 25) 2.5 (mt +> NCH2-CH3) 2.44 and 2 , 55 (2mt, -CII2-N (CH2CH3) 2) 2.62 (mt, H in 4) 2.74 and 3.10 (2dd, ^ CK2 in 15) 3.01 (mt, -S-Cg -) CH 3 3.69 and 3.89 (2d, 2: CH 2 in 17) 3.74 (mt, H in 26) 4.01 (d, J = 2.5, H in 27) 4.80 ( AB system, H in 13 and H in 14) 5.65 (d, H in 11) 6.60 (dd, H in 5) 7.71 (mt, ^ NH in 8) 25.80 (s, H in 20) l-Diethylamino-2 (R) -propanthiol can be prepared as follows.

To a suspension of 34.2 g of lithium aluminum hydride in 1600 ml of ethyl ether, 137.5 g of N, N-diethyl-2-mercapto (R) -propionamide dissolved in 500 ml of ethyl ether are added dropwise over 1 hour and 20 minutes. The reaction mixture is then kept at reflux for 2 hours and 30 minutes, after which it is cooled to a temperature near 0 ° C. Then, 40 ml of distilled water is added so that the temperature of the mixture does not exceed 20 ° C, and then 29.4 ml of 5N sodium hydroxide solution and then 133 ml of distilled water. The reaction mixture is filtered and the pH of the filtrate is adjusted to 8 by the addition of 70 ml of acetic acid. The resulting mixture is again filtered, rinsed with 3x 300 ml of ethyl ether, 5 and then the filtrate is dried over sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature near 30 ° C. A yellow oil is obtained which is purified by distillation under reduced pressure (1.3 kPa). There is thus obtained 101 g of 1-diethylamino-2 (R) -propanthiol in the form 10 of a colorless oil with a boiling point at 1.3 kPa of 55-56 ° C, [a] p ° - -37.1 ° C (c = 4.8, CH 3 OH).

N, N-Diethyl-2-mercapto- (R) -propionamide can be prepared as follows.

To 517 ml of a 5N sodium hydroxide solution maintained at 20 ° C, 105 g of N, N - diethyl-2-acetylthio- (R) -propionamide are added over 600 minutes in 600 ml of ethyl ether. The reaction mixture is stirred for 2 hours and 30 minutes at a temperature near 20 ° C. The aqueous base is separated by decantation.

The pH of the aqueous phase is then adjusted to 5-6 by the slow addition of 140 ml of acetic acid. The resulting mixture is extracted with 300 ml of dichloromethane and then 200 ml, dichloromethane and the organic bases are combined, dried over sodium sulfate, filtered and then concentrated to dryness under reduced pressure (2.7 kPa) at a temperature close to 30 ° C. There is thus obtained 78.6 g of N, N-diethyl-2-mercapto- (R) -propionamide in the form of a violet oil ([a] 2 ° = -21.1 "(c = 3.8) (CH 3 OH)).

N, N-diethyl-2-acetylthio- (R) -propionamide can be prepared as follows.

To a suspension of 72.6 g of the potassium salt of thiol-acetic acid in 300 ml of ethanol is added 88.3 g of N, N-diethyl-2-chloro (S) -propionamide dissolved in 150 ml of ethanol. The reaction mixture is then heated for 2 hours. a temperature 35 near 60 ° C, after which it is filtered and concentrated to dryness under reduced pressure (130 Pa) at a temperature near 50 ° C.

DK 169681 B1 16

The resulting residue is taken up and stirred in 500 ml of dichloromethane, after which it is washed with 300 ml of distilled water, 300 ml of a 10% aqueous potassium hydrogen carbonate solution and with 300 ml of distilled water. The organic phase is decanted, dried over sodium sulfate and 1 g of "3S" variety, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature near 30 ° C. The resulting residue is distilled off under reduced pressure (17 Pa). 105.5 g of N, N-diethyl-2-acetylthio- (R) -propionamide are thus obtained in the form of a pale yellow oil having a boiling point at 17 Pa of 105-107 ° C.

[.alpha.] D @ 20 = + 156 DEG (c = 18.1; CHCl3).

N, N-diethyl-2-chloro (S) -propionamide can be prepared as follows.

To a solution of 175.2 g of 2-chloro (S) propionyl chloride in 900 ml of chloroform maintained at 0 ° C is added 480 ml of diethylamine over 1 hour. To the reaction mixture is added 400 ml of distilled water and 300 ml of dichloromethane. The organic phase is decanted and washed with 300 ml of 2N hydrochloric acid and then 400 ml of distilled water. The organic phase is dried over sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature near 30 ° C.

The resulting residue is distilled off under reduced pressure (17 Pa). 161.9 g of N, N-diethyl-2-chloro (S) -propionamide in the form of a colorless oil having a boiling point are thus obtained.

Of) 25 at 17 Pa at 83-85 ° C. [α] p = + 39.7 ° (c = 10.4; CHCl 3).

2-Chloro- (R and S) -propionyl chloride can be prepared according to the method described by S-C.J. FU, S.M. BIRNBAUM and J.P. GREENSTEIN, J. Am. Chem. Soc. 76, 6054 (1954).

Example 4

To 2 g of 2- 6- [1-diethylamino-2 (S) -propyl] -thiopristiamycin IIB (isomer A) dissolved in 30 ml of methanol is added over 30 minutes at -60 ° C to 20 ml of an aqueous solution of 1.01 g of Oxone. The resulting suspension is stirred for 30 minutes at -60 ° C and then for 16 hours at -20 ° C. 0.2 g of "Oxone" dissolved in 5 ml of water is then added and then stirred for 30 minutes at t20 ° C. Dilute the reaction mixture with 100 ml of water, add 0.5 g of "NORIT® SX ULTRA" variety, stir for 15 minutes, filter on "Celite®" and then rinse with 3 times 5 ml of water. The aqueous phase 5 is extracted with 3 times 50 ml of ethyl acetate, adjusted to a pH of 7 by the addition of solid sodium hydrogen carbonate, saturated with sodium chloride and washed with 3 times 50 ml of dichloromethane.

The organic phases are combined, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C. In this way, a white meringue-like substance is obtained which is stirred in 20 ml of diethyl ether. The resulting solid is filtered off and dried under reduced pressure (270 Pa) at 20 ° C to give 1.5 g of 26- [1-diethyl-15-amino (S) -propyl] -sulfinylpristinamycin IIB in the form of a whitish solid (isomer A 2: 90%, isomer A 2: 5%), melting near 130 ° C and having the same NMR characteristics as the product of Example 5 below.

26- [1-Diethylamino-2 (S) propyl] -thiopristinamycin IIB (isomer A) can be prepared as follows.

To 10.5 g of pristinamycin IIA dissolved in 200 ml of a mixture of methylene chloride and methanol in the 50:50 volume ratio is added at -20 ° C under nitrogen 3.2 g of 1-diethylamino-2 (S) -propanthiol. After 70 hours of stirring at -20 ° C, the reaction mixture is concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C, after which up to 100 ml of ethyl acetate is taken up. The resulting solution is washed with 100 ml of a 0.2N aqueous solution of potassium hydrogen sulfate. The aqueous phase is decanted and washed with 3 times 100 ml of ethyl acetate.

The organic phase is washed with 50 ml of distilled water, then the aqueous phases are combined, adjusted to a pH of 7 by the addition of 2 g of solid sodium bicarbonate and washed with first 100 ml and then 2 times 50 ml of dichloromethane. The latter organic phases are combined, dried over 35 sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C. The resulting solid DK 169681 B1 18 is stirred in 150 ml of ethyl ether, then filtered and rinsed with 3 times 10 ml of ether. There is thus obtained 9.2 g of a beige solid which is recrystallized from 20 ml of acetonitrile. After filtration and drying at 20 ° C under reduced pressure (270 Pa), 2.3 g of 26- [1-diethylamino-2 (S) -propyl] -thiopristinamycin IIB (isomer A) is obtained as white crystals which melts near 128 ° C.

NMR Spectrum: 1.04 (t, -N (CH 2 CH 3) 2) 1.39 (d, 1 H 1.73 (S, -CH 3 in 33) 1.90 and 2.13 (2mt, CH2 in 25) 2.4-2.7 (mt, -CH2-N (CH2CH3) 2) 2.78 (mt, H in 4) 2.93 and 3.12 (2dd, ^ CE2 in 15) 20 3 3.52 (mt, H in 26) 3/84 (s, ^: CH2 in 17) 4.81 (s broad, H in 27) 4.49 (mt, -S-CH-) d, H i 13) 6.15 (d, H i 11) 6.30 (m,> NH i 8) 6.53 (dd, H i 5) 8.13 (s, H i 20) 30 l -Diethylamino-2 (S) -propanthiol can be prepared as follows.

To a suspension of 42.6 g of lithium aluminum hydride in 1500 ml of ethyl ether, 171 g of N, N-diethyl-2-mercapto (S) -propionamide dissolved in 500 ml of ethyl ether are added dropwise over 1 hour and 20 minutes. The reaction mixture is then kept under reflux for 2 hours and 30 minutes, after which it is cooled to a temperature near 0 ° C. Then 49.8 ml of distilled water is added in such a way that the temperature of the mixture does not exceed 20 ° C, and then 36.6 ml of 5N sodium hydroxide solution and 166 ml of distilled water. The reaction mixture is filtered and the pH of the filtrate is adjusted to 8 by the addition of 70 ml of acetic acid. The resulting mixture is filtered again and the filtrate is dried over sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature close to 30 ° C. A yellow oil is obtained which is purified by distillation under reduced pressure (1.3 kPa). 100 g of 1-diethylamino-2 (S) -propanthiol are thus obtained in the form of a colorless oil with a boiling point at 1.3 kPa of 55-56 ° C.

[Α] D = + 39.6 ° (c = 5.6, CH 3 OH).

N, N-Diethyl-2-mercapto- (S) -propionamide can be prepared as follows.

To 1100 ml of a 5N sodium hydroxide solution maintained at 20 ° C, 223 g of N, N - diethyl-2-acetylthio- (S) -propionamide are added in 1000 ml of ethyl ether over 30 minutes.

The reaction mixture is stirred for 20 hours at a temperature near 20 ° C. The aqueous phase is separated by decantation and washed with 250 ml of ethyl ether 3 times. The pH of the aqueous phase is then adjusted to 5 by the slow addition of 280 ml of acetic acid. The resulting mixture is extracted with 500 ml and then 2 times 250 ml of dichloromethane, and the combined organic phases are dried over sodium sulfate in the presence of carbon black, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature near 30 ° C. .

7.7 g of N, N-diethyl-2-mercapto- (S) -30-propionamide are obtained in the form of a violet oil ([α] β = + 20.7 ° (c = 3. CH 3 OH) ).

N, N-diethyl-2-acetylthio- (S) -propionamide can be prepared as follows.

To a suspension of 16 g of the potassium salt of thiol-diacetic acid in 70 ml of ethanol is added 20 g of N, N-diethyl-2-chloro - (R) -propionamide dissolved in 30 ml of ethanol. The reaction mixture DK 169681 B1 20 is then heated to a temperature near 55 ° C for 2 hours, then concentrated to dryness under reduced pressure (130 Pa) at a temperature near 60 ° C. The resulting residue is triturated with 300 ml of dichloromethane and then washed with 200 ml of distilled water. The organic phase is decanted, dried over sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature near 30 ° C.

The resulting residue is distilled off under reduced pressure (17 Pa). There are thus obtained 22 g of N, N-diethyl-2-acetylthio-10 - (S) -propionamide in the form of a pale yellow oil with a boiling point at 17 Pa of 105-107 ° C. [α] D = -169 ° (c = 10, CHCl 3).

N, N-diethyl-2-chloro (R) -propionamide can be prepared as follows.

To a solution maintained at 20 ° C of 87.7 g of 2-15 chloro (R) propionyl chloride in 600 ml of chloroform is added 153 g of diethylamine over 1 hour. The reaction mixture is washed with 500 ml of distilled water, 3 times 500 ml of 1N hydrochloric acid and then 500 ml of distilled water. The organic phase is dried over sodium sulfate, filtered and concentrated to dryness 20 under reduced pressure (2.7 kPa) at a temperature near 30 ° C.

The resulting residue is distilled off under reduced pressure (17 Pa). 43 g of N, N-diethyl-2-chloro (R) -propionamide are thus obtained in the form of a colorless oil having a boiling point at 17 Pa of 75-80 ° C. [α] D = -43.3 ° (c = 10; CHCl3).

25

Example 5

To a suspension of 63 g of pristinamycin 1¾ in 630 ml of methanol, 17.6 g of 1-diethylamino-2 (S) -propanthiol are added under nitrogen at -38 ° C over 30 minutes. After 20 hours of stirring, 150 ml of distilled water are added at -38 ° C and then in 30 minutes 410 ml of an aqueous solution of 33.6 g of Oxone. The resulting suspension is stirred for 1 hour at -38 ° C, 1.2 g of sodium thiosulfate is added, filtered at 20 ° C on a glass frit and then washed with 3 times 35 200 ml of distilled water. 20 g of "NORIT® SX ULTRA" black are added to the aqueous phase, stirred for 30 minutes and filtered on "Celite®" and the filtrate is washed with 3x 200 ml of distilled water. The solution is adjusted to a pH of 7 with 15 g of solid sodium bicarbonate, after which it is washed with 3 times 500 ml of dichloromethane. The organic phases are combined, dried (5) over sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C. 55 g of a beige marigold is obtained, which is stirred in 500 ml of ethyl ether. washing with 3 times 50 ml of ethyl ether and drying under reduced pressure (270 Pa) at 10 20 ° C gives 46.6 g of a white solid which is dissolved in 150 ml of a mixture of dichloromethane and methanol in a volume ratio 98: 2, This solution is purified by flash chromatography, eluting with a mixture of dichloromethane and methanol in a 98: 2 volume ratio and collecting 100 ml fractions. After concentration under reduced pressure (2.7 kPa) at At 30 DEG C., 36 g of a solid are dissolved in 400 ml of distilled water added with 6.5 g of potassium hydrogen sulfate. The solution is washed with 4 times 500 ml of ethyl acetate, adjusted to a pH of 7 by the addition of 5 g of solid sodium chloride. ium hydrogen carbonate and extracted thereto first with 200 ml and then twice with 150 ml of dichloromethane. The organic phases are combined, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C to give 33.8 g of a beige 25 meringue which is stirred in 300 ml of ethyl ether. After filtration, 29.3 g of 26- [i-diethylamino-2- (S) -propyl] -sulfinylpristinamycin lig (isomer A2) is isolated in the form of a pale yellow powder melting near 140 ° C.

NMR Spectrum (isomer A2): 1.05 (t, -N (CH2-CH3) 2) δ, 1.27 (d, OS jx) λ

H

1.74 (s, -CH3 in 33) 2.10 (mt, 1H of ^> CH2 in 25) DK 169681 B1 22 2.45-2.70 (mt, 1H of ^ CH2 in 25, 1H of> ch2-n (CH2CH3) 2, -N (CH2CH2) 2.80 (mt, H in 4) 2.88 (mt, 1H of I> CH2-N (CH2CH3) 2) 5 '2.90 and 3.15 (2dd, 2 ^ CH2 in 15) 3.06 (mt, -S-CH-) 4-1.0 ch3 3.41 (mt, Η in 26) 3.83 (S, ^ CH2 in 17) 10 4, 77 (s wide, Η in 27) 5.50 (d, H in 13) 6.20 (d, H in 11) 6.55 (mt, NH NH in 8) 6.63 (dd, H in 5) 8.11 (s, H i 20)

Example 6

To 50 g of 26- (2-diethylaminoethyl) -thiopristinamycin equilibrated in 200 ml of dichloromethane and 250 ml of distilled 20 water are added at 0 ° C over 20 minutes 30.7 g of Oxone dissolved in 140 ml of distilled water. After 10 minutes of stirring, the organic phase is decanted and the aqueous phase is washed with 100 ml of dichloromethane and then adjusted to a pH of 7 by the addition of 120 ml of saturated sodium hydrogen carbonate solution. The aqueous phase is washed with 5 times 100 ml of dichloromethane, adjusting the pH to 7 at each wash by adding sodium bicarbonate. The organic phases are combined, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 40 ° C to obtain 31 g of 26- (2-diethylaminoethyl) sulfinylpristinamycin equilibrium (isomer A2: 85 %, isomer 10%) in the form of a light beige powder having the same characteristics as the product of Example 1.

Example 7

To 2 g of 26- [1-diethylamino-2 (S) -propyl] -thiopristi- nam 169681 B1 23 namycin lig (isomer A) dissolved in 30 ml of ethanol and 20 ml of water are slowly added at 0 ° C 0.08 ml of concentrated sulfuric acid and then 0.9 g of "Oxone" dissolved in 10 ml of distilled water. The resulting solution is stirred for 4 hours at 20 ° C. Five solid sodium bicarbonate is then added to a pH of 4 and stirring is continued for 2 hours at 20 ° C. Then add 25 ml of distilled water and 2 g of "NORIT® SX ULTRA" - black. The mixture is filtered on "Celite®" and rinsed with 20 ml of dichloromethane, then the pH is adjusted to 7 10 with solid sodium bicarbonate.

The aqueous phase is decanted and washed with 2 times 25 ml of dichloromethane. The organic phases are dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 40 ° C.

In this way, 1.6 g of a beige solid containing 80% 26- [1-diethylamino-2 (S) -propyl] -sulfinylpristinamycin lig (isomer A2), 10% isomer A starting sulfide having the same characteristics as the product described in Example 4.

20

Example 8

To 5 g of 26- [1-diethylamino-2 (S) -propyl] -thiopristiamycin IIB (isomer A) dissolved in 20 ml of distilled water and 90 ml of methanol maintained at -5-30 ° C are added 0, 24 ml of concentrated sulfuric acid and then slowly 2.7 g of Oxone dissolved in 10 ml of distilled water. The resulting cloudy solution is stirred at -30 ° C for 1 hour. 0.6 g of "NORIT® SX ULTRA" variety is then added and the mixture is filtered on "Celite®". The filtrate is adjusted to a pH of 7 by the addition of solid sodium hydrogen carbonate and then washed with 3 times 50 ml of dichloromethane. The organic phases are combined, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C.

The resulting solid is stirred in a mixture of 20 ml of ethyl acetate and 70 ml of ethyl ether, then filtered and washed with 50 ml of ethyl ether. On this 4.5 g of DK 169681 B1 24 is obtained a light beige solid which is purified by flash chromatography, eluting with a mixture of ethyl acetate and methanol in the 90:10 volume ratio and collecting 15 ml fractions. After concentration to dryness of fractions 47-54 under reduced pressure (2.7 kPa) at 30 ° C, 1.7 g of 26- [1-diethylamino-2- (S) -propyl] -sulfinylpristinamycin is obtained (isomer A2) containing 10% starting sulfide and having the same characteristics as the product described in Example 4.

Example 9

To 25.5 g of 26- [2-diethylamino- (2S) -propyl] -thiopristiamycin dissolved in 350 ml of ethanol is added 200 ml of distilled water at -30 ° C and then slowly 15.1 g of Oxone dissolved in 70 ml distilled water. After stirring for 1 hour and 30 minutes at -30 ° C, 3.7 g of sodium thiosulfate dissolved in 20 ml of water is added. The reaction mixture is then poured onto 400 ml of distilled water and 200 ml of dichloromethane, then the pH is adjusted to 7%. The organic phase is decanted and the aqueous phase is washed with 200 x 200 ml of dichloromethane, the organic phases are combined, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C. In this way, 24 g of a pale yellow solid is obtained, which is purified by flash chromatography, eluting with a mixture of 25 chloroform and methanol in a 90:10 volume ratio. After concentration to dryness under reduced pressure (2.7 kPa) at 30 ° C of fractions 23-28 (volume 50 ml) is obtained a yellow marigold-like substance which is stirred in 100 ml of ethyl ether and then filtered and dried under reduced pressure (90 Pa) at 30 ° C. 3 g of 26- [2-diethylamino- (2 S) -propyl] -sulfinylpristinamycin IIB (isomer A2) in the form of a light yellow powder melting near 128 ° C.

NMR Spectrum: 1 1 .N (CH 5 CH) δ 1.1 (mt, -CH 2 CH 3 and S-CK 2 C 2 3 2) 1.77 (s, -CH 3 in 33) / 1 * CS 3 DK 169681 B1 2 (mt, 1H of -CH2- in 25) CH2-CH3 2.30-2.7 (mt, 1H of -CH2- in 25, -N,

Nsca2-cH3 5 ”1H of -SO-CH2-) 2.97 (mt, 1H of -SO-CH2-) 2.78 (mt, ^ CH- in 4) 2.92 and 3.10 (2dd, - CH2- i 15) 3.13 (mt, ^ CH- i 26) 3.50 (mt,> N-CH-) 3.81 (s, -CH2- 17) 4.77 (d, ^ CH - i 27) 5.52 (d, = CH i 13) 6.18 (d, = CH- i 11) 6.48 (mt, -NH- i 8) 6.57 (dd, = CH- i 5) 8.12 (S, = CH- in 20) 1- 6- [2-Diethylamino- (2S) -propyl] -thiopristinamycin IIB 20 (isomer A) can be prepared as follows.

To 31.5 g of pristinamycin 1¾ dissolved in 230 ml of methanol and 70 ml of chloroform placed under a nitrogen atmosphere are added at -40 ° C 10 g of 2-diethylamino- (S) -propanthiol dissolved in 50 ml of chloroform. After 4 days of stirring at 4-40 ° C, the mixture is concentrated to dryness under reduced pressure (2.7 kPa) at 35 ° C, after which the resulting solid is stirred in 500 ml of ethyl ether, filtered and again stirred in 300 ml. die-methyl ether. After filtration, the solid is dried at 90 Pa and 20 ° C, then purified by flash chromatography, eluting with a mixture of chloroform and methanol in the 90:10 volume ratio and 100 ml fractions are collected. After concentration to dryness under reduced pressure (2.7 kPa) at 30 ° C of fractions 18-30, 26.6 g of 26- [2-diethyl-amino (2S) -propyl] -thiopristinamycin IIB (isomer A) is obtained in the form 35 of a light yellow solid melting near 110 ° C.

NMR Spectrum: H (CH 2 CH 3) δ 1.05 (mt, ch 3 -ch 2 - and -s-ch 2 -c) VCH 3 1.72 (s, -CH 3 in 33) δ 1.97- 2.10 (mt, -CH2- in 25) ^ CH2-CH3 2.50 (mt,) ^ cn2-CE3 2.60, 2.87 and 3.04 (mt, -S -CH2-CH) , 77 (mt,> CH- in 4) 2.9 and 3.10 (2dd, -CH2- in 15) 3.35 (mt,> CH- in 26) 3.82 (S, -CH2- 17) 4.7 (d, = CH- i 27) 5.47 (d, = CH- i 13) 6.15 (d, = CH- i 11) 6.46 (m, -NH- ) 6.54 (dd, = CH- in 5) 2-Diethylamino- (S) -propanthiol can be prepared as follows.

To 39 g of 2-diethylamino (S) -propylisothiouronium dihydrochloride in 140 ml of diethyl ether are added 59.4 ml of 5N aqueous sodium hydroxide solution. After stirring for 25 minutes under a nitrogen atmosphere, the aqueous phase is decanted and washed with 3 times 150 ml of ethyl ether. The organic phases are combined, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C. The resulting liquid is distilled at 78 ° C under a pressure of 2.7 kPa to give 10.6 g of 2-diethylamino- (S) -propanethiol in the form of a fair-anhydrous liquid containing 6% 1-diethylamino- propanethiol. [α] 20 D = + 32 ± 0.6 ° (c = 0.943; ethanol).

2-Diethylamino- (S) -propylisothiouronium dihydrochloride can be prepared as follows.

To 18.2 g of thiourea dissolved in 80 ml of N, N-dimethylformamide is added at 130 ° C a solution of 45 g of 2-diethylamino-1-chloro (S) -propane hydrochloride in 150 ml of N, N-dimethyl 1 formamide. After 5 minutes at 130 ° C, the solution is cooled. The resulting crystals are filtered off, washed with ethyl ether and dried under reduced pressure (90 Pa) at 20 ° C. 39 g of 2-diethylamino (S) -propylisothouronium dihydrochloride are obtained in this form in the form of white crystals, melting at 209 ° C. [A] p = -2.6 "(c = 1; ethanol).

2-Diethylamino-1-chloro- (S) -propane hydrochloride can be prepared as follows.

To 110 ml of thionyl chloride is added at 2 ° C 50 g of 2-diethylamino (S) -propanol hydrochloride in small portions. The resulting solution is then heated to 60 ° C for 4 hours. The excess thionyl chloride is removed by distillation under reduced pressure. (90 kPa), then 250 ml of ethyl-ether are added to the residue, filtered off, the resulting solid is filtered off, rinsed with 300 ml of ethyl ether and then recrystallized from 150 ml of methyl isobutyl ketone. -diethylamino-1-chloro (S) -propane hydrochloride in the form of white crystals, 20 which melts at 103 ° C. [α] D = 0 "(c = 0.7; ethanol).

2-Diethylamino (S) -propanol hydrochloride can be prepared as follows.

To a suspension of 18.23 g of lithium aluminum hydride in 2000 ml of tetrahydrofuran, 48.66 g of L-acetyl-N-2-ethylamino (S) -propanol dissolved in 300 ml of tetrahydrofuran are added over 20 minutes. The reaction mixture is then refluxed for 4 hours, after which it is cooled to 0 ° C.

Slowly add 22 ml of distilled water, then 16 ml of 5N aqueous sodium hydroxide solution and then 72 ml of distilled water. After stirring for 1 hour at 20 ° C, the mixture is filtered on "Celite®". The filtrate is concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C, taken up with 300 ml of dichloromethane, dried over magnesium sulfate, filtered and then concentrated to dryness under reduced pressure (2.7 35 kPa) at 30 ° C. there is obtained 42.45 g of a light yellow liquid which is distilled under reduced pressure, 26.56 g of 2-diethylamino (S) -propanol in the form of a colorless liquid having a boiling point at 1 kPa of 55.5 ° C.

31.41 g of 2-diethylamino (S) -propanol hydrochloride are obtained in the form of a white solid melting at 98 ° C, after filtering off the crystals obtained and adding 40.5 ml of a 4.94N solution of hydrogen chloride in ethyl ether to the above product dissolved in 136 ml of acetone.

[α] p = +20.7 ° (c = 1; ethanol).

N-Acetyl-N-2-ethylamino- (S) -propanol can be prepared as follows.

To 9.15 g of N-2-ethylamino (S) -propanol dissolved in 100 ml of dichloromethane is added 13.7 ml of triethylamine at 0 ° C and 7.1 ml of acetyl chloride are added over 45 minutes. After the addition is complete, the temperature is allowed to rise to 15 to 20 ° C. The reaction mixture is diluted with 100 ml of water, adjusted to a pH of 9 by the addition of sodium carbonate and then washed with 2 times 200 ml of dichloromethane.

The organic phases are combined, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure 20 (2.7 kPa) at 30 ° C to give 9.54 g of a yellow liquid which is distilled at 108 ± 2 ° C. In this way, 6 g of a colorless liquid containing about 80% N-acetyl-N-2-ethylamino (S) -propanol and 20% of the corresponding diacetylated derivative are obtained. (used as such in the subsequent synthesis).

N-2-Ethylamino (S) -propanol can be prepared as follows.

To a suspension of 81 g of lithium aluminum hydride in 4000 ml of tetrahydrofuran is added at 0 DEG C. 227 g of 2-acetylamino-30 - (S) -propionic acid ethyl ester in 800 ml of tetrahydrofuran over 45 minutes. After the addition is complete, the temperature is allowed to rise to 20 ° C, after which the mixture is heated to reflux for 5 hours. After cooling to 0 ° C, 97 ml of distilled water is slowly added, then 72 ml of 5N aqueous sodium hydroxide solution and finally 320 ml of distilled water. The mixture is filtered at 20 ° C on "Celite®". The filtrate is concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C, taken up with 500 ml of dichloromethane, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at 30 ° C. ° C to give 143 g of a yellow liquid. After distillation under reduced pressure (460 Pa) at 57 ° C, 90.8 g of N-2-ethylamino (S) -propanol is obtained as a colorless oil.

[α] D = + 45.4 ° (c = 1.48? ethanol).

2-Acetylamino propionic acid ethyl ester can be prepared as described by J.P. WOLFF III et al., Biochemistry 2, 493 (1963).

Example 10 Proceeding in the same manner as described in Example 9 but starting from 9.1 g of 26- [N-methyl-N-cyclohexyl-1-amino-2- (S) -propyl] -thio-pristinamycin IIB (isomer A) and 5.1 g of Oxone are obtained 8.3 g of a beige solid which is purified by flash chromatography, eluting with a mixture of ethyl acetate and methanol in the ratio of 90:10 and 20 fractions of 20 ml are collected. After concentration to dryness under reduced pressure (2.7 kPa) at 30 ° C of fractions 32-70 and stirring the resulting solid in ethyl ether, 3.4 g of 26- [N-methyl-N-cyclohexyl-1-amino are obtained. -2- (S) -propyl] -sulfinyl-pristinamycin IIB (isomer A2) in the form of a pale yellow solid melting near 140 ° C.

NMR Spectrum (400 MHz) 1-1.3 and 1.5-1.85 (2mt, CH2 of cyclohexyl) 1> 27 <d 'οΑ / "2ΐ" 0' C-3 1.76 (s, -CH 3 in 33) 2.09 and 2.55 (2mt, 2 CH 2 in 25) 2.31 (s, N-CH 3) 2.37 (mt, CH cyclohexyl) DK 169681 B1 2.55 and 2.90 (2dd, N-CH 2 -) 2.78 (mt, H in 4) 2.92 and 3.16 (2dd, CH 2 in 15) 3 (mt, -S-CH 5) 0. 3.41 (d wide, H in 26) 3.83 (s, = = - CH 2 in 17) 4.78 (s, H in 27) 5.48 (d, H in 13) 6.18 (d, H in 11) 6.53 (dd, NH in 8) 6.60 (dd, H in 5) 8.10 (S, H in 20) 26- [N-methyl-N-cyclohexyl-1 -amino-2- (S) -propyl] -thio- -pristinamycin lig (isomer A) can be prepared as follows.

In the same manner as described in Example 4 for the preparation of 26- [1-diethylamino-2- (S) -propyl] -thio-pristinamycin 20 lig (isomer A), but by working in a mixture of methanol and chloroform in the volume ratio 70:30 and starting from 15 g of pristinamycin IIA and 5.9 g of N-methyl-N-cyclohexyl-1-amino-2- (S) -propanthiol are obtained after 48 hours at -30 ° C 10.6 g 26- [N-methyl-N-cyclohexyl-1-amino-2- (S) -propyl] -thiopristiamycin lig (isomer A) in the form of a beige solid melting near 160 ° C .

NMR Spectrum: 1-1.35 (mt, 1 CH 2 of cyclohexyl) 1.55-1.85 (mt, 1 CH 2 of cyclohexyl) 1.36 (d, -S-CH-NcC)% 3 72 (s, -CH 3 in 33) 1.90 and 2.10 (2 mt, 2 CH 2 in 25) 2.31 (s, 1 N-CH 3) 2.40 (mt, - 2 -CH of cyclohexyl) ) 2.5-2.6 (2dd,]> N-CH 2 ') DK 169681 B1 31 2.77 (mt, -H in 4) 2.92 and 3.13 (2dd, ^ CH2 in 15) 3, 05 (mt, -SCH-) 3.56 (mt, -H ~ i 26) 5 "3.82 (s, ^ CH 2 in 17) 4.82 (s wide, -H i 27) 5.48 (d, - H i 13) 6.17 (d, -H i 11) 6.48 (mt, ^ NH i 8) 6.54 (dd, -H i 5) 8.13 (s, -H i 20) N-Methyl -N-cyclohexyl-1-amino-2- (S) -propanthiol can be prepared as follows.

To a suspension of 15.7 g of lithium aluminum hydride in 850 ml of ethyl ether is added dropwise over 1 hour and 20 minutes 79.34 g of N-methyl-N-cyclohexyl-2-mercapto (S) -prop-ionamide dissolved in 200 ml of ethyl ether. The reaction mixture is then kept at reflux for 2 hours, after which it is cooled to a temperature near 0 ° C. Then, 45 ml of distilled water is added in such a way that the temperature of the mixture does not exceed 20 ° C, and then 45 ml of 5N sodium hydroxide solution and 150 ml of distilled water. The pH of the reaction mixture is adjusted to 8 by the addition of 40 ml of acetic acid. The resulting mixture is filtered, the filtrate is decanted and the aqueous phase is washed with 4x 400 ml of ethyl ether. The ether phases are combined, dried over sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature near 30 ° C. A yellow oil is obtained which is purified by distillation under reduced pressure (1.3 kPa). 37 g of N-methyl-N-cyclohexyl-1-amino-2- (S) -propanthiol is thus obtained in the form of a colorless oil with a boiling point at 0.2 kPa of 110-113 ° C.

[.alpha.] D @ 20 = + 27.2 DEG (c = 2.4, CHCl3).

N-methyl-N-cyclohexyl-2-mercapto- (S) -propionamide can be prepared as follows.

DK 169681 B1 32

To a solution of 89.76 g of N-methyl-N-cyclohexyl-2-acetylthio- (S) -propionamide in 85 ml of methanol is added at 13 ° C under nitrogen atmosphere 80 ml of ION sodium hydroxide solution. After stirring for 3 hours at 40 ° C, the mixture is concentrated to dryness under reduced pressure (2.7 kPa), then dissolved in 200 ml of distilled water and washed with 2 x 200 ml of ethyl ether. After decanting, the pH of the aqueous phase is adjusted to 5 by the slow addition of 30 ml of acetic acid. The resulting mixture is extracted with 3 times 10 100 ml of chloroform. The combined organic phases are dried over sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature near 30 ° C. 80.54 g of N-methyl-N-cyclohexyl-2-mercapto- (S) -propionamide in the form of an oil are thus obtained.

[Α] 25 D = + 24.5 ° (c = 1.1, CHCl 3).

N-Methyl-N-cyclohexyl-2-acetylthio- (S) -propionamide can be prepared as follows.

To a suspension of 56.2 g of the potassium salt of thiol-acetic acid in 200 ml of ethanol is added 80 g of N-methyl-N-cyclohexyl-2-chloro (R) -propionamide dissolved in 60 ml of ethanol. The reaction mixture is then heated to a temperature near 69 ° C for 45 minutes, then concentrated to dryness under reduced pressure (130 Pa) at 40 ° C. The resulting residue is triturated with 300 ml of dichloromethane, filtered and then washed with twice 300 ml of distilled water, 300 ml of 10% aqueous hydrogen carbonate solution and 300 ml of distilled water. The organic phase is decanted, dried over sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature near 30 ° C. 90.6 g of N-methyl-N-cyclohexyl-2-acetylthio- (S) -propionamide are thus obtained in the form of an orange-yellow oil.

[α] D 20 - -97.8 ° (c = 1, CHCl 3).

N-Methyl-N-cyclohexyl-2-chloro- (R) -propionamide can be prepared as follows.

To a solution maintained between -ί-4 and + 2 ° C of 70.7 g of 2-chloro (R) -propionyl chloride in 320 ml of chloroform DK 169681 B1 33 is added 294 ml of N-methyl-N-cyclohexylamine within 1 hour. The reaction mixture is washed with 250 ml distilled water, 3 times 250 ml 1N hydrochloric acid and 4 times 250 ml distilled water.

The organic phase is dried over sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature near 30 ° C. The resulting residue is distilled off under reduced pressure (135 Pa). 80.46 g of N-methyl-N-cyclohexyl-2-chloro (R) -propionamide are thus obtained in the form of a colorless oil having a boiling point at 0.13 kPa of 10 125 ° C.

20 [ar] * = -34 ° (without solvent).

Example 11 Proceeding in the same manner as described in Example 9, but starting from 14.4 g of 26- [1-diisopropylamino-2- (S) -propyl] -thiopristinamycin IIB (isomer A ) and 8.2 g of "Oxone" are obtained 14.1 g of a beige solid which is purified by flash chromatography, eluting with a mixture of ethyl acetate and methanol in the 90:10 volume ratio and collecting fraction 20 down to 20 ml. After concentration to dryness under reduced pressure (2.7 kPa) at 30 ° C of fractions 62-81, a yellow solid is obtained which is stirred in ethyl ether. After filtration and drying at 20 ° C under reduced pressure (270 kPa), 9.7 g of 26- [1-diisopropylamino-2- (S) -propyl] -sulfinyl-pristi-namycin IIB (isomer A2) is obtained a yellow solid which melts near 120 ° C.

NMR Spectrum: 1 (mt, -CH 3 isopropyl) 1.24 (d, S-CH-C 1-4 -N 2 C) δ CH 3 1.74 (s, -CH 3 in 33) 2 and 2.53 ( 2mt, ^ CH2 in 25) 2.67 and 2.84 (2mt, ^ N-CH2-) 2.77 (mt, -H in 4) 2.87 (mt, -S-CH2C) 169681 B1 34 2.85-3.05 (mt, -N (-CH = C) 2 + 1H of in 15) 3.15 (dd, 1H of ^ CH2 in 15) 3.37 (d wide, -H in 26) 3.81 (s, ^ .CH2 in 17) 5 ~ 4.65 (s wide, -H in 27) 5.51 (d, -H in 13) 6.20 (d, -H in 11) 6.64 (dd + mt, -H in 5 and 3 NH in 8) 8.11 (s, -H in 20) 10 2 6- [1-diisopropylamino-2- (S) -propyl] -thio -pristinamycin Hg (isomer A) can be prepared as follows.

Proceeding in the same manner as described in Example 4 for the preparation of 26- [1-diethylamino-2- (S) -propyl] -15-thio-pristinamycin IIB (isomer A), but working in a mixture of methanol and chloroform in the 80:20 volume ratio and starting from 20 g of pristinamycin IIÅ and 8.88 g of 1-diisopropylamino-2- (S) -propanethiol, a 48 hours at -30 ° C is obtained a light beige solid which is stirred in 150 ml of a mixture of ethyl ether and pentane in the 50:50 volume ratio. After filtration and drying at 20 ° C under reduced pressure (270 Pa), 16.8 g of 26- [1-diisopropylamino-2- (S) -propyl] -thio-pristinamycin IIB (isomer A) is obtained as a bright beige inherited solid, melting near 140 * 0.

NMR Spectrum (400 MHz) 1 (mt, -CH 3 isopropyl) 1.39 (d, -S-CH-CH 2 -l / N) CH 3 1.75 (s, -CH 3 in 33) 1, 93 and 2.15 (2mt, ^ CH2 in 25) 2.41 and 2.68 (2dd, ^ N-CH2-) 2.82 (mt, -H in 4) 2.96 (mt, -S- CHCC) DK 169681 B1 35 and 3.15 (2dd, ^ CH2 in 15) 3.05 (mt, -N (-CH <;) 2) 3.57 (mt, -H in 26) 3.88 (S, 2 CH 2 in 17) 5 '4.88 (s broad, -H in 27) 5.57 (d, -H in 13) 6.17 (d, -H in 11) 6.35 (mt, 2 >> NH i 8) 6.56 (dd, -H i 5) 8.15 (s, -H i 20) 1-Diisopropylamino-2 (S) -propanethiol can be prepared as follows.

1-Diisopropylamino-2 (S) -propanthiol can be prepared in the same manner as described in Example 10 for the preparation of N-methyl-N-cyclohexyl-1-amino-2 (S) -propanthol starting from 48 g N, N-diisopropyl-2-mercapto- (S) -propionamide and 10.16 g of lithium aluminum hydride. After reduced pressure (225 Pa) distillation, 24.1 g of 1-diisopropylamino-20 (S) -propanethiol is obtained in the form of a colorless oil having a boiling point at 235 Pa at 69 ° C.

[Α] p = + 24.2 ° (c = 1.3, CHCl3).

N, N-diisopropyl-2-mercapto- (S) -propionamide can be prepared in the same manner as described in Example 10 for the preparation of N-methyl-N-cyclohexyl-2-mercapto- (S) -propionamide, starting from 61.62 g of N, N-diisopropyl-2-acetyl-thio- (S) -propionamide and 55 ml of ION sodium hydroxide solution.

48 g of N, N-diisopropyl-2-mercapto- (S) -20 propionamide are obtained in the form of a red oil ([α] D = + 5.6 ° [c = 1, 30 CHCl3]), which is used as such for the subsequent turnover.

N, N-diisopropyl-2-acetylthio- (S) -propionamide can be prepared in the same manner as described in Example 10 for the preparation of N-methyl-N-cyclohexyl-2-acetylthio- (S) -35-propionamide, is taken from 46.8 g of the potassium salt of thiol acetic acid and 65.01 g of N, N-diisopropyl-2-chloro (R) -propionamide. 70.53 g of N, N-diisopropyl-2-acetylthio- (S) -propionamide are obtained in the form of yellow oil.

2 [α] D = -91.7 ° (c = 2, CHCl 3).

N, N-diisopropyl-2-chloro (R) -propionamide can be prepared in the same manner as described in Example 10 for the preparation of N-methyl-N-cyclohexyl-2-chloro (R) -propionamide starting from 73.6 g of 2-chloro (R) -propionyl chloride in 350 ml of chloroform and 166 ml of diisopropylamine to give 64.5 g of N, N-diisopropyl-2-chloro (R) -propionamide in the form of a colorless oil with a boiling point at 18 Pa at 71 ° C.

[Α] D = -73 ° (without solvent).

Together 1 icminas experiment: The yields of Examples 1-4 and 6-9 are compared to the yield obtained in Example 3 of EP-A1-191.662. Namely, the product of this example is the only one where the purity of the final product is comparable to the purity of the products prepared by the process of the invention.

20 The results are shown in the table below.

It can be seen from the comparison that in almost all cases the yields of isomer and A2 as well as the yields of isomer A2 are superior to very superior to the yields obtained by the process of EP 25 (no comparison has been made, e.g. 5 * s, using the process used there directly from pristinamycin 11¾, without isolation of the intermediate sulfide).

Thus, in addition to the ease of implementation, the process 30 of the invention has the advantage that the yields are significantly increased, both of isomer + A2 and of isomer A2 alone. It also has the importance of targeting turnover to form isomer A2.

35 DK 169681 B1 37

Example Starting compound Prepared for ^ Yield No. compound (g) (g)% 1 ~ 13 g 9.9 g 70.5% (isomer A) (isomer A2 B5% (A ·) + A2) isomer 10ί 10 «} 61 , 3% (A2) 2 3 g 2.4 g 72.5% (isomer A) (isomer A2 88% (Aj ♦ A2) isomer A1 6%) 67.9% (A2) 3 3.2 g 2.2 g 61.7% (A2) (isomer A (isomer A2 80%) 4 2 g 1.5 g 70.2% (isomer A) (isomer A2 90% (Aj + A2) • isomer * 1 10%) 66.5 % (A2) 6 50 g 31 g 57.5% (isomer A2 85% (A ·] + A2) isomer Al 10%) 51.5% (A2) 7 2 g 1.6 g 69.7% (isomer A) (isomer A2 90% (Αί * A2) isomer Ai 10%) 61.9% (A2) 8 6 g 1.7 g 24.9% (A2) (isomer A) (isomer ^ 2 90%) 9 25.5 g 11.3 g 43.4% (A2) (isomer * 2) DK 169681 B1 38

Example Starting Compound Prepared- Yield # * Compound (9) (9)% 10 9.1 g 3.4 g 35.7% (λ2) (isomer A) (isomer A2) 11 14.4 g 9.7 g 63 , 4% (A2) (isomer A) (isomer A2) EP 161 662 53.2 g 10.9 g ^ 32.2% (isomer A ^ 75%) (isomer A2 60% (A) * A2) Example 3 isomer Ai 15%) ♦ 5 g 28.2% (isomer A2) (A2 total)

Claims (1)

DK 169681 B1 39 Patent claims. Process for Preparation of Pristinamycin IIg - Derivatives of the General Formula I, wherein R is a) a nitrogen-containing heterocyclyl group of 4-7 ring members optionally containing a or several other heteroatoms selected from nitrogen, oxygen and sulfur in the form of sulfur oxide or sulfon, and optionally substituted by an alkyl group, or b) an alkyl chain of 2-4 carbon atoms substituted by one or two groups selected of phenyl, cycloalkylamino or N-alkyl-N-cycloalkylamino containing 3-6 ring link, alkylamino, dialkylamino and dialkylcarbamoyloxy (the alkyl moieties of the latter two groups optionally together with the nitrogen atom to which they are attached may form a saturated or unsaturated heterocyclic group of 4-7 ring members optionally containing another heteroatom selected from nitrogen, oxygen and sulfur in the form of sulfoxide or sulfone and optionally substituted with an alkyl group or is c) an alkyl chain of 2-4 carbon atoms substituted by one or more nitrogen-containing heterocyclic groups of 4-7 ring members optionally containing one or two other heteroatoms selected from nitrogen, oxygen and sulfur in the form of sulfoxide or sulfone, and optionally substituted with an alkyl group, said heterocyclic groups attached to the chain carrying them, via a carbon atom, or d) a [1-methyl-2 (S) -pyrrolidinyl] methyl group, at least one of the substituents carried by said alkyl chain is a nitrogen-containing substituent capable of forming salts, and the above alkyl groups being straight or branched and containing 1-10 carbon atoms, unless otherwise stated, in the form of their isomers or their mixtures, or their addition salts with acids, characterized in that a pristinamycin IIB derivative is oxidized by the triple salt [2KHS05 · KHS04 · K2S04] of the general formula II wherein R is as defined above except that when R contains a sulfur-containing heterocyclic group, the sulfur atom may also be in the form of a sulfide or a salt or protected derivative thereof, whereupon the resulting product is optionally separated into its isomers, the protecting group, if any, is removed and the resulting product is optionally converted to an addition salt thereof with an acid. 35