DE2949809A1 - (2S,5R,6S)-6-BROMINE-3,3-DIMETHYL-7-OXO-4-THIA-1-AZABICYCLO SQUARE BRACE TO 3.2.0 SQUARE BRACE TO HEPTAN-2-CARBONIC ACID-S,S-DIOXIDE, THEIR SALTS AND EASILY HYDROLYZABLE ESTERS - Google Patents

Description

29A9809

Bacteria sensitive to ß-lactam antibiotics can are known to develop resistance by forming a ß-lactamase enzyme. These enzymes catalyze the hydrolysis of the ß-lactam ring to form a ß-amino acid derivative that shows no antimicrobial effect. 10

A ß-lactamase enzyme is the RTEM enzyme. The structural gene of this enzyme is also in a plasmid. Apparently this enzyme is the most widespread ß-lactamase among the gram-negative intestinal bacteria; see Fisher et al., Annual Reports in Medicinal Chemistry, Vol. 13 (1978), p. 239 · The genetic information for the RTEM enzyme can be transformed into bacteria that are sensitive to β-lactam antibiotics, which makes them sensitive to the antibiotics become resistant. This ability to transform a resistance from organism to organism naturally poses serious problems. This is particularly evident in the development of ampicillin-resistant strains of N. gonorrhoeae. To eliminate the effects of ß-lactamases, two approaches have been taken so far. The first way is to synthesize new ß-lactam antibiotics that are stable to ß-lactamases. These efforts have been reasonably successful. However, the ß-lactamase-resistant antibiotics usually have a lower antibacterial activity than the analogous antibiotics, which are sensitive to ß-lactamase. The second possibility is to use a compound which inhibits the action of ß-lactamase enzymes on the ß-lactam ring. These ß-lactamase inhibitors can be administered together with ß-lactam antibiotics.

The invention is based on the object (2S, 5β »6S) -6-bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo ^ 3.2.Q7heptane-2-car-

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To create bonsäure-S, S-dioxides, their salts and easily hydrolyzable esters, which are inhibitors of the ß-lactamase enzyme RTEM. This object is achieved by the invention. The invention thus relates to the subject matter characterized in the preceding claims.

The compounds of the invention include both the 6 <x and 6β-forms of the compound, their salts and easily hydrolyzable Ester.

The (2S, 5R, 6S) -6 «. -Bromo-3,3-dimethy 1-7-OXO-4 - thia-1 -azabicyclo-Z3 «2.Q / heptane-2-carboxylic acid-S, S-dioxide, Its salts and easily hydrolyzable esters can be obtained by oxidation of the corresponding 6oc-bromopenicillanic acid can be obtained. The oxidation is carried out according to the usual methods for the oxidation of a sulfur atom. Specific examples of usable Oxidizing agents are m-chloroperbenzoic acid, hydrogen peroxide and sodium metaperiodate. The oxidation reaction can in an organic solvent such as ethyl acetate. Since the reaction temperature is not critical, the reaction is expediently carried out at room temperature.

6 oi-bromopenicillanic acid is known from J. Org. Chem., Vol. 27 (1962), p. 2668. It can be prepared by diazotizing a 6-aminopenicillanic acid at 0 to 2 ° C in dilute hydrobromic acid.

The (2S, 5R, 63) -6ß-bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo-Z3'2.0 / heptane-2-carboxylic acid-S, 3-dioxide ₁ its salts and Easily hydrolyzable esters can be made from 6,6-dibromopenicillanic acid. This compound is described, for example, in J. Chem. Soc, Part C, 1969, p. 2123.

In the process according to the invention, the carboxyl group of the 6,6-dibromopenicillanic acid must be protected from the reduction. This can be done by esterifying the acid with an alcohol

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achieve that sterically shields the carboxyl function. Suitable sterically shielding alcohols are benzhydryl alcohol and p-methoxybenzhydryl alcohol. The corresponding esters can selectively and without impairing the rest of the molecule be split off again. The benzhydryl ester can by reacting the free acid with diphenyldiazomethane in an organic solvent such as ethyl acetate. When converting the benzhydryl ester of 6,6-dibromopenicillanic acid the corresponding benzhydryl ester is precipitated with a reducing agent, preferably tri-n-butyltin hydride a 6β-bromopenicillanic acid.

The oxidation of the benzhydryl 6ß-bromopenicillanate provides the corresponding sulfone. The oxidation can be carried out by customary methods for oxidizing a sulfur atom will. Examples of oxidizing agents that can be used are m-chloroperbenzoic acid, hydrogen peroxide and potassium permanganate. The oxidation reaction can be carried out in an organic solvent such as ethyl acetate, or in a mixture of Water and a water-miscible organic solvent such as dioxane. As the reaction temperature is not critical, the oxidation reaction is expediently carried out at room temperature.

The treatment of the benzhydryl 6ß-bromopenicillanate with an acidic compound, preferably trifluoroacetic acid, provides 6ß-bromopenicillanic acid. The sodium salt is made by reacting this compound with sodium hydroxide solution obtain. The deuterated forms of water and sodium hydroxide can also be used.

Other methods of making the 6β fona are also available Disposal. For example, the (2S, 5R, 63) -6β-bromo-3,3 ^ 111 ^ 1 ^ 1-7-0X0-4-thia-i-azabicyclo / ^. 2. Q / heptane-2-carboxylic acid-S, S-dioxide produce by first the ester group of the benzhydryl 6ß-bromopenicillanate

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split off and then oxidized the acid obtained.

Salts, in particular physiologically compatible salts of (2S, 5R _t 6S) -6-bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo-Z3.2.o7heptane-2-carboxylic acid-S, 3 -Dioxides can be produced by customary methods. They are also ß-lactamase inhibitors. Specific examples of the salts are alkali and alkaline earth metal salts and amine salts.

The easily hydrolyzable esters of the compounds of the invention are produced by methods known per se. These compounds are also ß-lactamase inhibitors. Examples for ester groups which are easily hydrolyzable in vivo, i.e. under physiological conditions, have the structural part formula -CH (Y) -O-CO-alkyl, in which Y is a hydrogen atom or represents an alkyl radical. Specific examples are the acetoxymethyl, pivaloyloxymethyl, methoxymethyl and isobenzofuranyl groups.

The RTEM enzyme is a ß-lactamase enzyme that does hydrolysis of the lactam ring of ß-lactam antibiotics. The resulting derivative of the antibiotic is against the bacteria no longer active.

By simultaneous administration of a compound of the invention, its physiologically tolerated salt or easily hydrated Esters together with a ß-lactam antibiotic that is sensitive to the RT £ M enzyme can cause bacterial infections be combated more effectively. The compounds of the invention can be used simultaneously, i.e. as a combination preparation a ß-lactam antibiotic, or given separately.

In the combination preparation, the weight ratio of RTEM enzyme Inhibitor to ß-lactam antibiotic from about 1: 10 to 10: 1, preferably from about 1: 3 to 3: 1. The production these preparations for oral or parenteral administration

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can be done by customary methods.

Many of the known ß-lactarn antibiotics are sensitive versus the RTEM enzyme. According to Sykes et al., J. of Antimic. Chemotherapy, 2: 115 (1976), are most Penicillin antibiotics to varying degrees sensitive to the RTEM enzyme. Specific examples of such Penicillins are ampicillin, amoxicillin, penicillin V, penicillin G, carbenicillin and sulbenicillin. The cephalosporins are less sensitive to the RTEM enzyme. Examples for sensitive cephalosporins are cephaloridin and cephalothin.

The examples illustrate the invention. 15th

example 1

(23, 5R, 6S) -6 oC-Bromo-3,3-dimethyl-7-rOxo - ^ - thia-1-azabicyclo- / 3 · 2.0 / -heptane-2-carboxylic acid-S, S-dioxide

A solution of 20.00 g of m-chloroperbenzoic acid cooled to 0 to 5 ° C in 200 ml of ethyl acetate is mixed with a solution of 5.6 g (20 mmol) of (2S, 5R, 6S) -6-bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo / 3.2.0 / heptane-2-carboxylic acid added in 30 ml of ethyl acetate. After stirring for 15 to 18 hours at 26 ° C the reaction mixture is diluted with 3OO ml of ethyl acetate and twice with 150 ml of saturated aqueous sodium bicarbonate solution each time extracted. The aqueous extracts are combined and adjusted to pH with concentrated hydrochloric acid acidified by 2. The acidic solution is then quickly extracted three times with 200 ml of ethyl acetate each time. The ethyl acetate extracts are combined, dried over sodium sulfate and evaporated under reduced pressure at a slightly elevated temperature. A white solid remains, i.e. a mixture of the desired compound and m-chlorobenzoic acid. " By digesting three times with 1 liter of hexane each time, 5-7 g of a white pesticide are obtained, which is almost free

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C. 78 H 23 N 10 S. Br 60 30, 98 3, 11 4.48 10 , 27 25, 52. 30, 3, 4.62 , 32 25,

is from m-chlorobenzoic acid.

An analytically pure sample is prepared as follows: About 1 g of the product is dissolved in a mixture of chloroform and hexane and brought to the cloud point. To The supernatant is decanted from the separated oil for 1 hour. After standing for about 16 hours, crystals separate from the decanted solution and recrystallize from hot benzene will. The title connection is dated

F. 139 to 140 ⁰ C obtained.

IR absorption spectrum (KBr): 1793, 1780 (shoulder), 1755 cm NMR spectrum (acetone-d ₆ ): 1.50 (s, 3H), 1.60 (s, 3H), 4.48 (s, 1H), 5.16 (d, 1H, J = 2 Hz), 5.40 (d, 1H, J = 2 Hz). Analysis of CgH ₁₀

ber .:
found :

Example 2

(23,5R, 6S) -6β-Bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo- / 3.2.07heptane-2-carboxylic acid-3,3-dioxide

a) (2S, 5R, 6S) -6,6-dibromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo / 3.2.07heptane-2-carboxylic acid benzhydryl ester A solution of 23.24 g (64.7 mmol) of (2S, 5R, 63) -6,6-dibromo-3,3-dimethyl-7-oxO "4-thia-1-azabicyclo / 3.2.07heptane-2 -carboxylic acid in 400 ml of ethyl acetate is stirred dropwise with a solution of 12.57 g (64.7 mmol) of diphenyldiazomethane added in 100 ml of ethyl acetate. During the addition, nitrogen escapes and the reaction mixture is at about 16 hours Let stand room temperature. Thereafter, the reaction mixture with saturated aqueous sodium bicarbonate solution and Washed water, dried over sodium sulfate and taken under evaporated under reduced pressure. A foam remains which solidifies after the addition of diethyl ether. The product is chromatographed on 500 g of silica gel and from diethyl ether

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recrystallized. There are 17.13 g of the title compound, melting at 128 to 13O ⁰ C was obtained.

IR absorption spectrum (moMull): 1800, 1745 cm " ¹ The analysis C ₂₁ H _ig N0 SBr ₂ ;

C H N Br

calc .: 48.02 3.65 2.67 30.43

found: 48.27 3.59 2.62 30.40.

b) (2S, 5R, 6S) -6β-Bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclop. 2.07h ept an-2-carboxylic acid benzhy dry reads a solution of 2 g (3.81 mmol) (2S, 5R, 6S) -6,6-dibromo-3,3-dimethyl-7-oxo-4-thia- 1-azabicyclc / 3.2.07heptane-2-carboxylic acid in 50 ml of anhydrous dichloromethane is cooled in an ice bath under nitrogen as protective gas and with 1.10 ml (4.17 mmol) of tri-n-butyltin hydride were added. The mixture will Stirred at 0 to 5 ° C for 4 hours. The reaction mixture is then left to stand at 12 ° C. for about 16 hours. Then will the solvent is distilled off under reduced pressure. An oil remains which is chromatographed on 125 g of silica gel will. The product is eluted with dichloromethane. The eluate is evaporated. There are 856 mg of the title compound as an oil obtain.

IR Absorption Spectrum (CHCl ₂ ): 1790, 1750cm. Nuclear Magnetic Resonance Spectrum (CDCl ₃ ): S, 1.23 (S, CH ₃ ), 1.62 (S, CH ₃ ), 4.6

(S, H ₃ ), 5.25 (d, J = 4 Hz, H ₅ ), 5.55 (d, J = 4 Hz, H ₅ ).

c) (2S, 5R, 6S) -6ß-Bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyolo-3 «2.07heptane-2-carboxylic acid S, S-dioxide-benzhydryl ester

A solution of 456 mg (1.02 mmol) of (2S, 5R, 6S) -6β-Bromo-3,3-dimethyl ^ -oxo ^ -thia-i-azabicyclo / B ^ .Oy'heptan ^ -carboxylic acid benzhydryl ester in 3O ml dichloromethane is added with 477 mg (2.35 mmol) of m-chloroperbenzoic acid (purity 85%) were added. The mixture is stirred for 24 hours at room temperature, then washed with saturated aqueous sodium bicarbonate solution and water and dried over sodium sulfate. Then that will

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Solvent distilled off under reduced pressure. An oil remains which is purified by preparative thin-layer chromatography on silica gel with dichloromethane as the mobile phase. The title compound is obtained as a foam. IR Absorption Spectrum (CHCl ₃ ): 1815, 1755 cm " ¹ .

Nuclear Magnetic Resonance Spectrum (CD _{3 COCD 3} ): ä, 1.2 (S, CH ₃ ), 1.6 (S, CH), 4.75 (S, H ₃ ), 5.27 (d, J = 4 , 5, H ₅ ), 5.87 (d, J = 4.5, H ₅ ). Analysis C ₂₁ H ₂₀ NO ₅ SBr;

C H N Br

calc .: 52.72 4.21 2.93 16.71

Found: 52.83 4.07 2.70 16.43.

d) (2S, 5R, 6S) -6ß-Bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo / 3.2.o7heptane-2-carboxylic acid-S, S-dioxide A solution of 316 mg (0.661 mmol) (2S, 5R, 6S) -6β-Bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo / ^ 2. Q / heptane-2-carboxylic acid-S, S-dioxide benzhydryl ester and 361 μl (330 mmol) of anhydrous anisole in 5 ml of anhydrous dichloromethane is cooled to -15 to -2O ° C under nitrogen as a protective gas and treated with 1 ml of anhydrous trifluoroacetic acid. The mixture is stirred for 3 hours with cooling. The solvent is then evaporated under reduced pressure and without external supply of heat. Residual trifluoroacetic acid is distilled off by joint distillation with benzene under reduced pressure at room temperature. The residue obtained is digested once with a mixture of diethyl ether and pentane and three times with diethyl ether. 153 mg of the title compound with a melting point of 128 ° to 133 ° C. remain behind.
IR Absorption Spectrum (KBr): 1810, I76O (COOH), I710 cm " ¹ .

Nuclear Magnetic Resonance Spectrum (CDCl ₃ -CD ₂ OD): i, 1.47 (S, CH ₃ ), 1.63 (S, CH), 4.15 (S, H ₃ ), 5.02 (d, J = 4.5 Hz, H ₅ ), 5.62 (d, J = 4.5 Hz, Hg). The analysis CgH ₁₀ NO ₅ SBr;

C H N Br calc .: 30.78 3.23 4.49 25.60

found: 31.36 3.13 4.03 26.21.

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Example3

The benzhydryl ester obtained in Example 2 (b) is at -10 to -2O ° C in a mixture of anhydrous trifluoroacetic acid and dichloromethane (volume ratio 1: 5) cleaved under acidic conditions. The 6β-bromopenicillanic acid obtained is in Deuterium oxide (DpO) dissolved with the equivalent amount of sodium deuteroxide (NaDO). It becomes a solution of the relative Obtained sodium salt of 6ß-bromopenicillanic acid.

Example 4

(23 _> 5Ri6S) -6ß-Bromo-3,3-d.iniethyl-7-oxo-4-thia-1-azabicyclo- / 3.2.0 / heptane-2-carboxylic acid Na salt

a) (2S, 5R> 6S) -6ß-Bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo / 3.2.Q7heptane-2-carboxylic acid

A solution of 386 mg (0.865 mmol) (2S, 5R, 6S) -6ß-Bromo-3,3-dimethyl ^ -oxo ^ -thia-i-azabicyclo / B ^ .oyheptan ^ -carboxylic acid benzhydryl ester (Example 2 (b)) ) and 472 / ul (432 mmol) of anhydrous anisole in 5 ml of anhydrous methylene chloride is cooled to -10 to -20 ⁰ C. 1 ml of anhydrous trifluoroacetic acid is then added to the solution. The mixture is then stirred for about 3 3/4 hours at -10 to -20 ⁰ C. Thereafter, the solvent is distilled off under reduced pressure and residual trifluoroacetic acid is separated off by joint distillation under reduced pressure. The residue obtained is digested twice with methylene chloride and three times with benzene. 180 mg of the title compound are obtained as a solid.
IR absorption spectrum (gauze): 1800, 1755, 1705 cm.

Nuclear Magnetic Resonance Spectrum (CDCl ₅ -CD ₅ OD): / 4.45 (H ₅ ), 5.37 and 5.57 (H ₅ and

Hc, J = 4 Hz).
6 'ice '

b) (2S ₁ 5R, 6S) -6ß-bromo-3,3-dimet hy l ^ -oxo ^ -thia-i-azabicyclo ^ · 2. pyheptane ^ -carboxylic acid Na salt The in example 4 (a) The free acid obtained is dissolved in deuterium oxide with the equivalent amount of sodium deuteroxide (NaDO). The title compound is obtained.

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Claims (11)

VOSSIUS VOSSIUS TAUCHN ER · HEUNEMANN · RAUH SIEBERTSTRASSE A ■ 8OOO MUNICH 86 · PHONE: (O89) 47 4O7S CABLE: B EN ZOLPATENT MÖNCHEN - TELEX β-2 9 46 3 VOPAT D u.z .: P 4-25 (Vo / H) December 11, 1979 Case: V-968,538-S ER SQUIBB & SONS, INC.
Princeton, NJ, V.St.A. - "(2S, 5R, 63) -6-bromo-3,3-dimethy 1-7-OXO-4-- thia-1-azabicyclo- / 3.2.o7heptane-2-carboxylic acid-S, 3-dioxide, their salts and easily hydrolyzable esters " Priority: December 11, 1978, V.St.A., No. 968 538 December 11, 1978, V.St.A., No. 968 539 Claims 20 JU ¹ (2S, 5R, 6S) -6-bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo-Z3.2.Q / heptane-2-carboxylic acid-S, S-dioxide, their salts and easily hydrolyzable esters. 2. (2S, 5R, 6S) -6cv-3rom- or -6ß-Bromo-3,3-dimethyl-7-oxo-4 ~ thia-1-azabicyclo / 3 · 2.O / heptane-2-carboxylic acid-S, S-dioxide, its salts and easily hydrolyzable esters. 3. Process for the preparation of (2S, 5R, 6S) -6ß-bromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo / ^. 2.Q / heptane-2-carbon- acid esters, characterized in that one (2S, 5R, 6S) -6,6-dibromo-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo- / 3-2.0 / heptane-2-carboxylic acid ester with tri-n-butyltin hydride in a solvent and in a non-oxidizing one Brings atmosphere to implementation. 030025/0806 ORIGINAL INSPECTED -2- 29498Q9 4. The method according to claim 3 »characterized in that the reaction is carried out at temperatures of 0 to 5 ⁰ C in dichloromethane and under nitrogen as a protective gas. 5. The method of claim 3 »characterized in that there is used as starting Sesterheim the benzhydryl ester or the p-Methoxybenzhydrylester. 6. The method of claim 3 »characterized in that one converts the obtained ester with an acid-reacting compound in the 6i? -Brompenicillansäure and optionally transforming this compound with sodium hydroxide solution to the sodium salt. 7. The method according to claim 6, characterized in that the acidic compound is anhydrous trifluoroacetic acid used. 8. The method according to claim 5 »characterized in that one treated the benzhydryl ester obtained with an oxidizing agent and an acidic compound and in the S, S-dioxide is transferred and this compound is optionally converted into the sodium salt with sodium hydroxide solution. 9. The method according to claim 8, characterized in that one m-chloroperbenzoic acid is used as the oxidizing agent and anhydrous trifluoroacetic acid is used as the acidic compound . 10. Use of the compounds according to one of claims 1 or 2 as ß-lactamase enzyme RTEM inhibitors. 11. Medicinal product, characterized by a content of a ß-lactam antibiotic which is sensitive to the ß-lactamase enzyme RTEM as well as (2S, 5R, 63) -6 06-bromo- or -6ß-bromo-3,3-dimethyl l-7-oxo-4-thia-1-azabicyclo / 3.2.0 / heptane-2- 030025/0805 carboxylic acid-SjS-dioxide, its salt or easily hydrolyzable Ester. 030025/0805