DE1816824A1 - Process for the preparation of acylamino compounds - Google Patents

Description

CIBA AKTIENGESELLSCHAFT, BASEL (SWITZERLAND)

Case 63 ^ 7 / 1-3
Germany

Process for the production of acylamino compounds "

The invention relates to the production of new therapeutically effective derivatives of 7-amino-cephalosporanic acid of formula I.

R -SH ₀ CC-HN-CH-

COOH 909831/1531

in which R _{1 is} a heterocyclyl radical with at least 2 heteroatoms which is bonded to the sulfur atom of the mercaptoeaetyl group with a carbon atom lying directly between 2 heteroatoms, and in which R _{2 is} a free or esterified by a carboxylic acid hydroxyl group in which ester oxygen atoms can be replaced by sulfur atoms, an optionally N-substituted carbamoyloxy group in which oxygen atoms can be replaced by sulfur, or is a quaternary amino group, and its optionally internal salts.

R ₁ is especially a monocyclic heterocyclyl ■ rest, especially a 5- or 6-membered heterocyclyl radical which contains, as hetero atoms nitrogen, oxygen "or sulfur atoms. The ring is preferably at least partially unsaturated. Thus R, for example, imidazolyl (2 ), Imidazolinyl (2), 0xazolyl (2), 0xazolinyl (2), thiazolyl (2), thiazolinyl (2), 1,2,4-triazolyl (3), 1,3, £ -dithiazolyl (2), 1 , 2,4-thiadiazolyl (3), pyrimidyl (2), 1,2,4-triazinyl (2), 1,3,5-triazinyl (2), i, 3-thiazinyl (2), 1,3, 4-thiadiazinyl (2), 1,3,5-thiadiazinyl (4), 1,3,4-oxadiazinyl (2), 1,3,5- dioxazinyl (2). The radicals can also be substituted, in particular by "lower alkyl, lower alkoxy, hydroxy *: mercapto * halogen

and / or contain a fused-on benzene ring.

An esterified hydroxyl group Rg, in which oxygen atoms can be replaced by sulfur, is derived from a carboxylic acid and is preferably the acetoxy

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group or, for example, by lower alkyl, lower alkoxy or lower alkyl mercapto radicals, halogen atoms or the nitro group, mono- or dicyclic arylcarbonyloxy or thiocarbonyloxy, arylcarbonyl mercapto or thiocarbonyl mercapto group, in particular the benzoylmer'captogruppe. Further examples for R ₂ are:

a) a carbamoyloxy group of the formula

-O-CO-NH-R,,

wherein R., an aliphatic, aromatic, araliphatic or heterocyclic radical, especially an unsubstituted or substituted one, preferably by one or more Lower alkoxy groups or halogen atoms substituted, straight or branched lower alkyl radical, such as methyl, ethyl, but above all the β-chloroethyl radical is; or

b) a thiocarbamoyl mercapto group of the formula

S-CS-N

where R, has the meaning given above and R ^ for what-

o is hydrogen or R; or
to

o> c) a quaternary amino group in which the quaternary

«^ Ary nitrogen atom, for example, part of an aromatic ring, such as ^w a quinoline, isoquinoline or pyrimidine ring, especially an unsubstituted or substituted pyridine ring, for example of the formula

■ - 4 -

N ~ - / \

where R (- for hydrogen or one or more lower alkyl, Lower alkoxycarboiyl, carbamoyl or carboxyl groups or one or more halogen atoms.

The salts of the new compounds are metal salts, especially those of therapeutically applicable alkali or Alkaline earth metals, such as sodium, potassium, ammonium, calcium, or salts with organic bases, e.g. triethylamine, N-ethyl-piperidine, Dibenzylethylenediamine, procaine. If the group Rp is basic, internal salts can form.

The new compounds have a particularly good antibacterial effect. They are against both gram-positive and especially against gram-negative bacteria effective, e.g. against Staphylococcus aureus penicillin resilstent, Escherichia coli, Klebsiella pneumoniae, Salmonella typhosa and Bacterium proteus, as shown in animal experiments, e.g. on mice. You can therefore fight it of infections caused by such microorganisms, as well as feed additives, for the preservation of food or as a disinfectant. Connections are particularly valuable

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genes in which R, Iroidazolyl (2) ,; Imiüazölinsrl. (2.); .Thiazolyl (-2), 2? Hi.s2olinyl (2) or thiazolyl (2) uncL.Rg- die; Acetoxy group pe, the ß-Chloräthylcarbamoyl- or an unsubstituted or, as indicated above, is substituted pyridinio.

The new connections are preserved when one

a) a compound of the formula II S

Zr-m-öE CH CH ₂

G- N / C-CH.-Rp II

o / Y  ²

COOH

where Z is a halogenoacetyl radical such as fluorine, chlorine, iodine or, above all, bromoacetyl radical and R _{0 represents} the above

f has given meaning with a mercapto compound of

formula

R ₁ -SH,

in which R, which has the abovementioned meaning, reacts, or b) a compound of the formula II in which Z is hydrogen, _{acylated by the group R - S-CH 2} -CQ- and, if desired, compounds obtained with free or converted into one another by a carboxylic acid-esterified hydroxyl group R ^ and, if desired, in compounds obtained in which R _{2 represents} a hydroxyl group esterified by a carboxylic acid, in the oxygen atoms

909831 / 1S31

181682Λ - 6 -

Sulfur atoms can be replaced, this group stands in a manner known per se by an optionally N-substituted one Carbamoyloxy group in which oxygen atoms can be replaced by sulfur, or by a quaternary Amino group replaced, and optionally, if desired, the compounds obtained in their metal, such as alkali or alkaline earth metal salts, or salts with ammonia or organic bases, or the salts obtained forms free carboxylic acids or optionally internal salts. :

The reaction of the compound II, in which Z is a haloacetyl group, with the mercapto compound is preferably in an inert solvent for the compound such as methylene chloride, chloroform, carbon tetrachloride, Tetrahydrofuran, dioxane, dimethylformamide or acetonitrile and in the presence of a hydrogen halide binding agent, e.g. a weak inorganic base such as an alkali carbonate, bicarbonate or acetate or a tertiary one Amine, preferably ethyldiisopropylamine (Hünig base). The reaction takes place at room temperature

ό took place for a few hours; she can, if desired, also with

££ carried out at a lower or slightly higher temperature "** ■> will.

_ω . The acylation of the compound II, - in which Z is hydrogen, is carried out in the manner known for the acylation of amino acids, for example by means of an acid halide,

especially acid chloride, or acid azide or an acid anhydride, especially a mixed anhydride, for example a mixed anhydride formed with monoesterified carbonic acid, pivalic acid or trichloroacetic acid, or with the free acid itself in the presence of a condensing agent such as a carbodiimide, e.g. dicyclohexylcarbodiimide. The acylation of the compound II can also be carried out in such a way that the compound II, in which Z is hydrogen, is first silylated or stannylated, the silylation or stannylation product with the acid or a reactive acid derivative which represents the group R ₁ - S-CHp-CO- contains, acylated and any silyl or stanny groups present are split off by alcohol or water, cf., for example, British patent 1,073,530 and Dutch application 67.17107.

It is preferred to use those starting materials which lead to the particularly effective end products mentioned.

The cephalosporin derivatives used as starting materials are known or can be used according to known ones Process are produced.

The invention also relates to those execution forms of the method, after which one on at any stage of the process as an intermediate

• 909831/1531

constant connection goes out and the missing procedural steps carries out, or the process is terminated at any stage, or in which the starting materials are below forms the reaction conditions, or in which the reaction components are optionally present in the form of their salts.

The neural compounds can be used as remedies, z.3. in the form of pharmaceutical preparations, use. These contain the compounds mixed with one for pharmaceutical organic or inorganic suitable for enteral, topical or parenteral application, solid or liquid carrier material. For the formation of the same such substances come into question, those with the new Compounds do not react, such as water, gelatin, lactose, starch, stearyl alcohol, magnesium stearate, talc, vegetable oils, benzyl alcohols, rubber, propylene glycol, Polyalkylene glycols, petrolatum, cholesterol, or other known excipients. Pharmaceutical preparations can e.g. as tablets, coated tablets, ointments, creams, capsules or in liquid form as solutions, suspensions or Emulsions are present. If necessary, they are sterilized and / or contain auxiliary substances such as preservatives and stabilizers. Wetting or emulsifying agents, solubilizers or salts to change the osmotic pressure or

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Buffer. They can also contain other therapeutically valuable substances. The preparations are made according to the usual Methods received.

The invention is described in the following examples. The temperatures are given in degrees Celsius.

The abbreviation 7-ACA is used below for 7-aminocephalosporanic acid.

In thin layer chromatography on silica gel plates the following systems are used: System 52 = n-butanol-glacial acetic acid-water (75: 7.5: 21) System 101 A = n-butanol-pyrldine-glacial acetic acid-water

(42: 24: 4: 30).

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Example 1 :

3 # 93 g (10 mmol) of bromoacetyl ^ ACA are in 20 ml Methylene chloride with the addition of 3.45 ml (20 mmol) of N-ethyldiisopropylamine solved. A solution of 1.37 ß (12: mMoj) 2-mercapto-1-methylimidazole in 20 ml of methylene chloride is added. It is rinsed with 10 ml of methylene chloride.

After 7 hours, the solvent is removed in vacuo and the resulting oil is admixed with 10 ml of methanol and 1.65 ml of glacial acetic acid, after which it crystallizes almost completely after a short time. It is filtered off with suction, washed with methanol, dried and the practically pure 7- [l -. * Is obtained. Methyl-imidazolyl- (2) -mercapto-acetylamino-cephalosporanic acid. P. 177-179 ⁰ (decomp.) In evacuated capillary.

The substance is converted into the crystalline sodium salt as follows: 2.9 g are slurried in 60 ml of methanol, and 3 × 75 ml of 3-methyl-methanoic / sodium a-ethylhexanoate are added. When stirring and briefly heating in a water bath at 30 °, temporary dissolution and then crystallization of the sodium salt occurs. The mixture is concentrated in vacuo to about 15 ml in volume, left to stand at -20 ° for one hour, suction filtered, washed with a mixture of tetrahydrofuran and methanol (1: 1) and dried in vacuo. The sodium salt is easily soluble in water and has an [a] ^ ° = + 107 ° + 1 ° (c = 1, H ₃ O), the UV spectrum (in

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Water) shows a maximum at Z = 256 πιμ; (£, = 12,800). P. 150 - 154 ° (with decomposition).

Rf ₅₂ . 0.04; Rf _{101 A} = 0.35.

Example 2:

3 * 93 g (10 mmol) of bromoetyl-7-ACA are dissolved in 15 ml of methylene chloride with the addition of 3 * 45 ml (20 mmol) of N, N-diisopropylethylamine. ^1st solved. A solution is added which has been prepared by dissolving 1.23 β (12 mmol) of 2-mercaptoimidazoline in 20 ml of dimethylformamide and then diluting it with 10 ml of methylene chloride. It is rinsed with 5 ml of methylene chloride.

After 6 hours, the methylene chloride is removed in a water jet vacuum, then the dimethylformamide is distilled off ^{as completely as possible under a vacuum of about 0.3 mm uride:} "a temperature of 30 °. 10.7 g of thick oil result. By adding obtained from 100 ml of tetrahydrofuran

o> one a Precipitat that is intensely kneaded. Man ο

^ Filtered and the residue was triturate with a further 50 ml of tetrahydrofuran ~ *, filtered off and washed with little tetrahydro- furan ^ and then with hexane. After drying in vacuo, 6.75 g of a yellowish powder are obtained.

4.5 g of this crude product are dissolved in 13.5 ml Dimethylformamide and diluted with 13.5 ml of methanol. the

Solution is shaken with a spatula tip "Norit" (activated charcoal) at room temperature for 5 minutes, then filtered through a layer of "Hyflo Supercel" (diatomaceous earth) and the filter washed with 6 ml of a mixture of dimethylformamide and methanol (1: 1). Piltrate and washing liquid are combined and placed in front with 165 ml of tetrahydrofuran, whereupon a light yellow precipitate forms. It is left to stand at -20 for 1 hour. It is then filtered off with suction and the residue is washed very well in succession with a mixture of methanol ~ tetrahydrofuran (1: 4.5), tetrahydrofuran and finally with hexane. After drying, the 7-imidazolinyl (2) -mercaptoacetylamino 3-cephalosporanic acid is obtained as an almost colorless amorphous powder which is uniform in the thin-layer chromatogram. Rf ₅₂ = 0.05; Rf _{101 A} - 0.25. Melting point (evacuated capillary): 213-216 ° (decomposition). The optical rotation [a3 _ß = + 143 ° ±.-2 ° (in water, c = 0, ^ 5).

Example 3:

3.93 g of bromoacetyl-7-ACA are dissolved in 27 ml of methylene chloride with the addition of 3.45 ml of N, N-diisopropylethylamine solved. A solution is added which is obtained by dissolving 1 * 43 g of 2-mercaptothiazoline in 2 ml of dimethylformamide and subsequent dilution with 8 ml methylene chloride.

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has been presented 1st. Rinse with 10 ml of methylene chloride.

After 7 hours, the solvent is first in a water jet vacuum and then at approx. 0.3 nm (water bath 30) removed. 7 g of a viscous oil are obtained.

6.3 g of this crude product are in 27 ml of 10 # Potassium hydrogen phosphate solution added. By adding 0.3 ml of 2-n. Soda solution adjusts the pH to 5. then the solution is poured into a separating funnel and first with 55 ml of methylene chloride, then with 90 ml of ethyl acetate extracted. The organic phases are washed twice in succession with 45 ml of phosphate buffer at pH 5 each time and discarded. Then all aqueous phases are combined and covered with 180 ml of ethyl acetate, whereupon one through Addition of 11 ml 1-n. Hydrochloric acid adjusts the pH to 2.9 and saturates the aqueous phase with sodium chloride. It will be fine shaken, the phases separated and the aqueous phase extracted twice with 90 ml of ethyl acetate each time. the organic phases are successively saturated with 45 ml Washed brine, getrock-Q net with sodium sulfate, combined and evaporated to dryness in vacuo. Of the

oo amorphous residue crystallizes after the addition of 6 ml of one w

^ Mixture of ethyl acetate and tetrahydrofuran (9: 1) almost completely through. The crystals (1.76 g) are filtered off with suction and ⁱ ~ washed with ethyl acetate. They are cleaned by dissolving them in 15 ml of tetrahydrofuran - using a spatula

Shakes pointed "Norit" at room temperature, ^{filtered through a "Hyflo-Supercel n} -6chicht, the filter washes with 5 ml of tetrahydrofuran, the solvent removed in vacuo and the amorphous residue from 5 ml of a mixture of ethyl acetate and tetrahydrofuran (9: 1) The crystals are filtered off with suction, washed with ethyl acetate and dried in a high vacuum. The pure ftthiazolinyl- (2) -mercaptoacetylamino] -cephalosporanic acid is obtained with a melting point (evacuated capillary): I3I - I38 (decomposition). For the preparation of the sodium salt, 1, 6 g of the acid are suspended in 32 ml of methanol and 2.1 ml of 3% methanolic acid sodium ethylhexanoate are added The solution thus obtained is concentrated in vacuo to a volume of 8-10 ml and left for about 2 1/2 hours . standing -20 ° is then filtered off with suction and the crystals of the sodium salt are washed successively with 12 ml methanol-ethyl acetate (1: 1), washed 5 ml of ethyl acetate and hexane, melting point of the sodium salt. 185 - I89 ⁰ ( with decomposition). The optical rotation [a] _D = + 120 ° + 1 ° (in water, c = 1). Rf ₅₂ = 0.26; Rf _{101 A} - 0.55.

₅₂ = 0.26; Rf _{101 A}

Example 4:

3 * 93 ß bromoacetyl-7-ACA are in 20 ml of methylene chloride with the addition of 3 * ^ 5 ml of diisopropylethylamine

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solves. A solution is added which is obtained by dissolving 1.22 g of 3-mercapto-1,2,2-triazole in 10 ml of dimethylformamide and diluting with 10 ml of methylene chloride. It is rinsed with 10 ml of methylene chloride.

After 7 hours, the solvent is first distilled off in a water jet and then at approx. 0.2 mm (V / water bath 30 40 °) and 10.5 g of a brownish oil remain. This crude product is taken up in 20 ml of a 10 # strength potassium dihydrogen phosphate solution. By adding 0.9 ml of 2-n. Soda solution, the pH is adjusted to 5. Then the solution in the separating funnel is extracted successively with 80 ml of methylene chloride and 1 to 10 ml of ethyl acetate. The organic phases are washed twice with 20 ml of phosphate buffer of pH 5 each time. The combined aqueous phases are then covered with 300 ml of ethyl acetate and the pH is adjusted to 2.9 by adding 1.0 ml of concentrated hydrochloric acid. This creates a ... colorless pre-eiv; ·. ¹ V ■;. ·· - pitat that fulfills both phases. The entire contents of the vision funnel are through one. Glass filter filtered off with suction ■ and the residue washed with 100 ml of ethyl acetate. Repeated trituration of this precipitate with tetrahydrofuran (approx. 150 ml) allows the largely pure and crystallizing 7- [l2,4-triazoly3r (3) -mereaptoacetylaminoj-cephalosporanic acid to be dissolved out. The piltrate comprising both phases is separated off and further raw crystals are obtained therefrom. 9Q9831 / 1531

BATH ORIGINAL

2.5 g of the combined raw crystals are dissolved in 30 ml of a mixture of tetrahydrofuran and methanol (l-: l), shaken briefly with a spatula tip "Norit" at room temperature, filtered through a layer of "Hyflo Supercel" and the Pilte-r with 3 ml of the above mixture and 5 ml of methanol were rewashed. The piltrate is then reduced to approx. 5 ni in a vacuum. ^! . a, whereupon it crystallized immediately after inoculation, Melting point: 149 - I5I ⁰ (dec).

1.55 g of the acid are suspended in 50 ml of methanol and 1.85 ml of 3% metiianoli.SLties; .Natri. ^And inäthy.lhexano'at are added. The clear solution is reduced to approx. 10-12 ml in vacuo 2.5 ml of tetrahydrofuran are added and the mixture is left to stand at -20 for 2 hours, then suction filtered and the residue washed successively with methanol-tetrahydrofuran (1: 1), tetrahydrofuran, ethyl acetate and hexane .

20 Optical rotation of the pure sodium salt [a] _D

+ 113 ° + 1 °; melting point: 220 - 230 ⁰ (with decomposition).

Rf ₅₂ = 0.21; Rf _{101 A} - 0.5.

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Example 5:

'. 39 * 3 ^m S bromoethyl-7-ACA are dissolved in 0.2 ml of methylene chloride with the addition of 25.8 mg of N, N-diisopropylethylamine. A solution is added which has been prepared by dissolving 17.4 mg of 2-mercapto-4,5-dimethyl-thiazole in 0.05 ml of dimethylformamide and diluting with 0.15 ml of methylene chloride. It is rinsed with 0.05 ml of methylene chloride. .

After 7 hours the solvent is removed under 0.3 mm vacuum (water bath 30 °) and the resulting oil is worked up in the manner described in Example 3. 7- [4 _i 5-dimethyl-thiazolyl- (2) -mercapto-acetylamino] -cephalosporanic acid is obtained. Rf ^ ₂ = 0.35; ^R.

Example 6:

393 mg (1.0 mmol) of bromoacetyl-7-ACA are in 2.0 ml of methylene chloride with the addition of 258 mg of Ν, Ν-diisopropylethylamine solved. A solution is added which is obtained by dissolving 180 mg of 2,5-dimereapto-1,3,4-thiadiazole in 0.5 ml Dimethylformamide and diluting with 2.2 ml of methylene chloride. It is rinsed with 1 ml of methylene chloride. 909831/1531

After 6 hours, the solvent is first in a water jet vacuum and then at approx. 0.2 mm (water bath 30 °) distilled off and there remain about 1.1 g of yellowish oil. This crude product is shown in 10 ml of a potassium dihydrogen phosphate solution added and by adding 0.3 wl 2-n. Brought soda solution to pH 5. Then im Separating funnel extracted successively with 20 ml of methylene chloride and 30 ml of ethyl acetate. Washes the organic phases one after the other with twice 10 ml of phosphate buffer pH 5 · Then the aqueous phases are combined and with 40 ml Ethyl acetate layered. By adding 2.7 ml of 1-n. Hydrochloric acid is adjusted to pH 2.9 and the aqueous phase is saturated with sodium chloride. After vigorous shaking, the phases separated and the aqueous solution extracted twice more with 20 ml of ethyl acetate each time. The 3 organic phases are washed successively with 8 ml of saturated sodium chloride solution. It is then dried with sodium sulfate and filtered and evaporated to dryness in vacuo. The residue crystallizes from a mixture of ethyl acetate-tetrahydrofuran (9: 1).

Melting point of 7- [5-mercapto-1,3 * 4-thiadiazolyl- (2) -mercapto-acetylamino] -cephalosporic acid: 130-142 with decomposition (evacuated capillary). Rfcp ⁼ ° * 36; Rf ₁₀ -,. = 0.6.

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Example 7:

1 * 97 S bromoacetyl-7-ACA are dissolved in 5 ml of methylene chloride with the addition of 1.29 S (1 * 73 ml) N, N-diisopropylethylamine solved. A solution is added which is obtained by dissolving 0.975 g of 2-mercapto-benzimidazole in 5 ml of dimethylformamide and diluting with 10 ml of methylene chloride. It is rinsed with 5 ml of methylene chloride.

After 5 hours - the solvent is first in a water jet vacuum and then at approx. 0.2 mm (water bath 30) distilled off. The resulting light brown oil is taken up in 10 ml of a 10% potassium dihydrogen phosphate solution and by adding 0.5 ml of 2-n '. Soda solution adjusted the pH to 5. Then in the separating funnel twice with each 100 ml of ethyl acetate are extracted and the extracts are washed twice with 5 ml of phosphate buffer pH 5 each time. The watery Phases are combined, covered with a layer of 250 ml of ethyl acetate and the pH is adjusted by adding 9 ml of 1-n. Hydrochloric acid to 2.5

cd set. This forms a resinous precipitate, the ο

^ consists largely of the desired product. It will - * separated and kneaded with hexane, from which ^ tenen solid residue is the 7- [benzimidazo] yl- (2) -mercapto- - * Acetylaminoj-cephalosporanic acid extracted using tetrahydrofuran. The two-phase piltrate obtained after separating off the precipitate is saturated with sodium chloride. After strong

Tiger shaking, the organic phase is separated off, the aqueous solution is extracted twice more with 100 ml of ethyl acetate each time and the 3 aqueous phases are washed successively with 10 ml of saturated sodium chloride solution, dried with sodium sulfate, filtered and the combined organic solutions are concentrated in vacuo to a volume of approx. 10 ml, whereby most of the 7 .- [benzimidazolyl- (2) -mercaptoacetylamino] -cephalosporanic acid crystallizes. Melting point: 135 ₂ = 0.35; R 6

Example 8

11 * 75 g of 3- (Besacetqxymethyl) -3-benzoylthiomethyl-7-bromoacetylamino-cephalosporanic acid are added with the addition of 8.75 ml of N-ethyl-diisopropylamine in 75 ml of dimethylformamide solved. A solution of 3.58 g of 2-mercaptothiazoline is added in 25 ml of dimethylformamide and leaves with exclusion of light stand at room temperature. After 20 hours, the solvent is for the most part in vacuo (0.5-1 mm HGj distilled off and the resulting oily residue was added to 250 ml of phosphate buffer pH β and 250 ml of ethyl acetate. The pH of the aqueous phase is then adjusted to 6 and the phases are separated. The organic phase is discarded. the

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aqueous phase is adjusted to pH 2.3 with hydrochloric acid and saturated with sodium chloride. It is extracted successively with 1000, 250 and 250 ml of ethyl acetate. The ethyl acetate extracts are washed with saturated sodium chloride solution, dried with sodium sulfate and filtered through a column of 75 g of silica gel. The combined filtrates give a dry residue of 11 g. This is chromatographed on a column of 520 g of silica gel with chloroform and chloroform-acetone mixtures. The chloroform-acetone (9: l) eluates deliver J, 6 g of amorphous 3- (Desacetoxymethyl) -3-t> enzoylthiomethyl-7- [thiazolinyl (2) -mercaptoaeetylamino] cephalosporanic acid, which by means of sodium-a-äthylh «Xanoate into the crystalline sodium salt of the formula

-N
VS-CH ₀ -CO-NH-

can be convicted. Rf 52 = 0.50. In the UV spectrum in water there are maxima at } i = 240 nm (£ = 19,200) and ^ = 274 nm (£ = 20,000).

The 3- (Desacetoxymethy ^^ - benzoylthiomethyl ^ -bromacetylaminocephalosporan-

90983171531

acid, it can be prepared as follows: A solution of 17.5 g of 3- (desacetoxymethyl) -3-benzoylthiomethyl) -7-aminocephalosporanic acid (cf. Belgian Patent 650 444) and 12.5 ml of triethylamine in 1 l of dimethylformamide . is for one hour in a well-stirred and kept at -I3 to -15 ° solution of 9.2 ml of bromoacetyl bromide added dropwise to 100 ml of methylene chloride (nitrogen atmosphere) The temperature is left slowly for 1/2 hour rise to 10 and hold it there for another half an hour. Then most of the solvents are in the Distilled off vacuum at 0.5-1 mm Hg against a condenser of dry ice acetone. The oily product is on a Poured phosphate buffer of pH 6 and shaken with 1 l of ethyl acetate. At the boundary of the two phases arises a precipitate which is separated off by filtration or centrifugation. Then adjust the pH of the aqueous Phase to 2, saturate it with sodium chloride and separate the organic phase. The aqueous phase is with 6OO and 400 ml of ethyl acetate after-extracted. After washing with saturated saline, the organic Phases dried over sodium sulfate and filtered successively through a column of 100 mg silica gel. The filtrates are concentrated to dryness in vacuo, 30 ml of ethanol are added to the residue and the mixture is crystallized at -20 °.

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7 * 8 g of 3- (deacetoxymethyl) -3-benzoylthiomethyl-7-bromoacetylamino-cephalosporanic acid with a melting point of 137-138 ^{0 are obtained} . Rf 52 = 0.55. The sodium salt shows in the UV spectrum in water:

2 max 243 nm (£ = ΐβ'8θθ) and 275 nm (£ = 20 ^f 600).

[a] _j = -47 + 1 ° (c = Ij in 0.1 mol. sodium bicarbonate acetone (l: l).

Example 9

g of 3- (I ⁾ esacetoxymethyl) -3-benzoylthiomethyl-7- [thiazolinyl (2) -mercaptoacetylamino3-cephalosporanic acid (sodium salt) are dissolved in 35 ml of pyridine and then 35 ml of dioxane are added. Then 20.9 rol of a 40 # strength mercury perchlorate solution are added and the mixture is left to react at 45 ° for 45 minutes with vigorous stirring (nitrogen atmosphere). It is cooled, mixed with 11.1 ml of thiobenzoic acid and shaken for 5 minutes. The solvents are distilled off in vacuo and a solution of the residue in 140 ml of water is filtered off through "Celite". The Piltrat is washed successively with 9 ° ^m l of toluene, 2 times with 55 ml "Amberlite" LA-2 in 115 ml

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Toluene and washed twice with 90 ml of toluene each time. Then filter ^; the aqueous phase is passed through a column of 8.5 ml "Sephadex" CM C-25 (H ⁺ form), 34 ml "Alox", 8.5 ml "Zeo-Karb" 226 (H ⁺ -form), 34 ml of "Älox", 8.5 ml of "Dowex-1" (acetate form) and 8.5 ml of "Sephadex" CM C-25 (H ⁺ form). "Celite", organic phases and the column are re-extracted twice with 30 ml of water each time, and the column is also eluted with a further 200 ml of water. The combined eluates are concentrated in vacuo, a small amount of precipitate is removed by filtration and evaporated to dryness. The residue is digested with 10 ml of alcohol and gives the pure 3- (desacetoxymethyl) -3- .. pyridinlomethyl-7- [thiazolinyl (2 ") - mercaptoacetylamino] -

cephalosporanic acid. [a] _D = + 4l +1 (c = 1 in water); UV spectrum in water: max 257 πιμ (β = 13 * 000). Rf IOIA = 0.18.

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Example 10

2.95 g of the sodium salt of 7- [thiazolinyl (2) -mercaptoacetylaminol-cephalosporanic acid (cf. Example 3) are dissolved in 150 ml of water. The solution is warmed to 37 ° and 0.7 ml of 0.1-n. Sodium hydroxide solution adjusted to pH 7.5. A suspension of 60 mg of acetylesterase (from Bacillus subtilis ATCC 6633, cf. English Patent 1 080 9 ° 4) in 2 ml of water is then added and the acetic acid formed is continuously neutralized with the aid of an automatic titrator with O _f ln. Sodium hydroxide solution (adjustment to pH 7.3) · The reaction has ended after 4 hours. The pH is adjusted to 6.5, the solution is filtered through a G4 glass frit and lyophilized. 3.18 g of a yellowish resin of the sodium salt of 7- [thiazolinyl (2) -mercaptoacetylamino JO-desacetyl-cephalosporanic acid result.

This crude product is directly in a mixture of 30 ml of absolute dimethylformamide and 8.4 ml of triethylamine dissolved, treated with 5.05 ml of ß-chloroethyl isocyanate and stirred for half an hour at room temperature. The solvent is then removed in a high vacuum and the resinous, brown residue is triturated three times with each ISO ml of absolute ether. The ether-insoluble residue .Is taken up in 75 ml of 10 # phosphorus buffer of pH 6.7 and covered with 500 ml of ethyl acetate. By adding 30 ml of 2-n. Hydrochloric acid is adjusted to the pH of the aqueous phase

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to 2.5 a. After vigorous shaking, the phases are separated and the aqueous phase is then extracted again with 300 and 200 ml of ethyl acetate. The organic phases are washed twice with 50 ml of saturated sodium chloride solution each time, dried with sodium sulfate and evaporated to dryness in vacuo. The resulting resin crystallizes after adding a little acetone. It is removed and the crystals are washed with a mixture of equal parts of acetone and ether. The raw crystals can be further purified by mixing a suspension in 18 ml of methanol with 1.2 ml of a 3-m. methanolic solution of sodium-a-ethylhexanoptt is added, the mixture is clarified with a little "Hyflo" (diatomaceous earth), concentrated in vacuo to approx. 8 ml and at -15 ° C the pure sodium salt of O-deacetyl-O-iß-chloroethylcarbamoyl) 7- [thiazolinyl (2) -mercaptoacetylamino] -cephalosporanic acid crystallized out. In the ultraviolet spectrum it has a _Μαν at 255 nm with6 = IO85O (in water). The optical rotation Ea] ₀ 1st +99 + 1 . It shows the following values in the thin-layer chromatogram: for system 52 = 0.35,

"" 101A = 0.55 *

and in the system ethyl acetate-pyridine-glacial acetic acid-water (62: 21: 6: 11) Rf = 0.34.

909831/1531

Claims (1)

"'- 27-. Claims: 1. Process for the preparation of new derivatives of 7-aminocephalosporanic acid, characterized in that compounds of the formula I ⁰ p ■ / R ₁ -SH ₂ OO-HN-CH CH COOH wherein R _{1 is} a heterocyclyl radical with at least 2 hetero atoms, which is bonded to the sulfur atom of the mercaptoacetyl group with a carbon atom lying directly between 2 heteroatoms, and "where Rp is a free or esterified by a carboxylic acid hydroxyl group, in the ester oxygen atoms by sulfur atoms can be replaced, a given! if N-substituted CarDamoyloxygruppe, in which oxygen atoms can be replaced by sulfur, or is a quaternary amino group and its, if appropriate, internal salts are prepared by a) a compound of the formula II Z-NH-CH CH CH ₂ C, C-CH ₂ -R ₂ II 609831/1531 ⁰ ^ [ COOH -28 - wherein Z stands for a haloacetyl radical and R. has the meaning given above, with a mercapto compound of formula R ₁ -SH. , wherein R _{1 has} the abovementioned meaning, converts, or b) a compound of the formula II in which Z is hydrogen, _{acylated by the group R 1} -S-CHp-CO- and, if desired, compounds obtained with free or through a carboxylic acid-esterified hydroxyl group R _{2 is converted} into one another and, if desired, in compounds obtained in which Rp is a hydroxyl group esterified by a carboxylic acid, in which oxygen atoms can be replaced by sulfur atoms, this group is known per se Way by an optionally N-substituted carbamoyloxy group in which oxygen atoms are replaced by sulfur can be, or replaced by a quaternary amino group, and optionally, if desired, the compounds obtained converted into their metal, such as alkali or alkaline earth metal salts, or salts with ammonia or organic bases or the free carboxylic acids from the salts obtained ^ω or optionally forms internal salts. ^ 2. The method according to claim 1, characterized in that that a compound of the formula II, wherein Z is stands the bromoacetyl radical, with a mercapto compound of Formula FL-SH implements. 3. The method according to claims 1 or 2, characterized characterized in that the reaction with the mercapto compound of the formula R, -SH in the presence of Ν, Ν-diisopropylethylamine is made. 4. The method according to claims 1-3 * thereby characterized in that it is reacted with a mercapto compound of the formula R, -SH, in which R, is a monocyclic one 5 - 6-membered heterocyclyl radical, the as heteroatoms. Contains nitrogen, oxygen or sulfur atoms., And unsubstituted or by lower alkyl or alkoxy groups or hydroxyl groups and / or a condensed one Contains benzene ring. 5. The method according to claim 4, characterized in that the heterocyclic radical is at least partially unsaturated is. 6. Process according to claims 1-5, characterized in that one reacts with a mercapto compound of the formula R ^ -SH, in which R is an iridazolyl, inidazolinyl, thiazolyl, thiazolinyl, lode * »ptazolyl radical. 9 0 9 831/15 31 -3P - 7. Process according to claims 1-6, characterized in that compounds of the formula I are prepared, where Rg stands for the acetoxy group. 8 # Method according to claims 1 - "6, characterized in that that compounds of the formula I in which there is a pyridinio group are prepared. 9 · Process according to claims 1-6, characterized in that compounds of the formula I in which is a lower alkyl-carbamoyloxy group. 10. The method according to claims 1-6, characterized in that compounds of the formula I in which R _{2 is} a carbamoyloxy group of the formula -0-CO-NH-R, wherein R, a, pen manufactures by one or more halogen atoms or Niederalkoxygrup- is substituted lower alkyl · 11 · Process according to claims 1-6, characterized in that compounds of the formula I in which R_ is a carbamoyloxy group of the formula 909831/1531 -0-CO-NH-R ₃ 3Λ is where R 1 is substituted by one or more chlorine atoms Is lower alkyl. 12. The method according to claims 1-6, characterized in that that compounds of the formula I in which R 1 is the fJ-chloroethylcarbamoyloxy group are prepared. 13. Process for the production of .of- • f- (heterocyclyl-mer captoacetylaminoj-cephalosporanic acids as described in the examples. 14. Compounds of the formula I. 0 I. IL-a-H-O-O-HN-CH COOH in which R _{1 is} a heterocyclyl radical with at least 2 heteroatoms which is bonded to the sulfur atom of the mercaptoacetyl group with a carbon atom lying directly between 2 heteroatoms, and in which R "is a free or esterified by a carboxylic acid hydroxyl group in which ester oxygen atoms can be replaced by sulfur atoms, an optionally N-substituted carbamoyloxy group in which oxygen atoms can be replaced by sulfur, or is a quaternary amino group, and its salts. 909131/1531 -32 - ¹ ■ ' 15 .. compounds of formula I COOH wherein R stands for a monocyclic 5-6 membered heterocyclyl radical which contains nitrogen, oxygen w or sulfur atoms as heteroatoms and is unsubstituted or substituted by lower alkyl or alkoxy groups or hydroxyl groups and / or contains a fused benzene ring, and Rg contains the in Claim 14 has given meaning, and their. Salts. . Is 16. Compounds of formula I wherein R. is a monocyclic 5-6 membered heterocyclyl radical which corresponds as hetero atoms nitrogen, oxygen or sulfur atoms, which maintains and is unsubstituted or substituted by lower alkyl or alkoxy groups or hydroxyl groups and the Rg Is acetoxy group, and its salts. 17 · Compounds of the formula I in which R 1 is a monocyclic 5-6 membered heterocyclyl radical which contains nitrogen, oxygen or sulfur atoms as heteroatoms and is unsubstituted or by lower alkyl or. Alkoxy groups or hydroxyl groups is substituted and R is a pyridinio group, and their salts. 909831/1531 • -33 - 18. Compounds of the formula I in which R 1 is a monocyclic 5-6 membered heterocyclyl radical which contains nitrogen, oxygen or sulfur atoms as heteroatoms and is unsubstituted or substituted by lower alkyl or alkoxy groups or hydroxyl groups and R _{p is} a lower alkyl carbamoyloxy group and its salts. 19 · Compounds of the formula I in which R, is a monocyclic 5-6 membered heterocyclyl radical which is used as Heteroatoms contains nitrogen, oxygen or sulfur atoms and is unsubstituted or by lower alkyl or Alkoxy groups or hydroxyl groups substituted 1st and R « a carbamoyloxy group of the formula -O-CO-NH-R ,. is where R-. one through one or more lower alkoxy groups or halogen atoms is a substituted lower alkyl radical, and their salts. 20. Compounds of the formula I in which R _{1 is} a monooyclic 5-6 membered heterooyclyl radical which contains nitrogen, oxygen or sulfur atoms as heteroatoms and is unsubstituted or substituted by lower alkyl or alicoxy groups or hydroxyl groups and R _g the ß- Chlorethylcarbamoyloxygruppe 1st, and its salts. 90983171531 21. Compounds of formula I in which R ^ is imidazolyl. (2) -, imidazolinyl (2) -, Thiazolyl®- ,. Thiazolinyl (2) - or Triazolyl (2) group and R, the acetoxy, ß-Chloräthylcarbamojäoxy or Is pyridiniogroup, and its salts, 22. 7- [1-Methyl-imidazolyl (2) -mercaptoacetylamino] -cephalosporanic acid and their salts. 23 «7 ~ [Imidazolinyl (2) -mercaptoacetylamino] -cephalosporanic acid and their salts. 24 x 7- [Thiazounyl (2) -mereaptoacetylamino J-cephalosporanic acid and their salts. 25 · 7-U # 2,4-triazolyl (3) -mercaptoacetylamino] -ce- ■ phalosporanic acid and its salts. 26 · The compounds described in the examples. 27 · Pharmaceutical preparations containing cephalosporanic acid derivatives according to claims 14 - 26 · 28 «Animal feed or additives containing compounds according to claims 14-26. 909831/1531