CZ347597A3 - Esters of carbapenem compounds and pharmaceutical composition containing thereof - Google Patents

Abstract

A compound of formula (I), wherein R is selected from the group consisting of isobutyryloxymethyl, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl, and benzoyloxymethyl, is useful in the treatment of bacterial infections.

Description

The present invention relates to novel antibacterial compounds, to a process for their preparation, to pharmaceutical and veterinary compositions containing them and to their use in antibacterial therapy.

^: λ

BACKGROUND OF THE INVENTION

Carbapenems such as imipenem, compound of formula (A): HO Η H *

(A) have a strong and broad spectrum of antibacterial activity (see US 159550357 and US 4,194,047, Merek and Co.). However, these carbapenems tend to be hydrolyzed by the enzyme renin dihydropeptidase 1 (DHP1), and this limits their use in chemotherapy. In the case of imipenem, this problem can be avoided by co-administration of a DHP-1 inhibitor.

DHP-1 stability can be obtained by chemical modification of the substituent, as is the case with the meropenem of formula (B):

(B)

-2 (see Shih D. H. et al., Heterocycles, 1984, 21, 29 and Sunagawa M. et al., J. Antibiotics, 1990, 43, 519). This treatment has recently been extended to a β-aminoalkyl substituent (see EP 0 433 759, BristolMeyers Squibb).

An alternative approach for obtaining improved stability to DHP-1 utilizes carbon-substituted carbapenems such as 2-aryl, 2-heteroaryl and 2-heteroaromatic carbapenems (US 4,543,257, 4,260,627, US 4,962,101, US 4,978) 659, EP 0 14 493, EP 0 414 489, EP 0 010 316 and EP 0 030 032, Gauge and Co.) and 2- (substituted) methylcarbapenem (Schmidt et al., J. Anubiotics, 41, 1988, 780). _

UK Patent 1,593,524, Gauge & Co. discloses a series of five-membered heteroaromatic carbapenem derivatives, including diazolyl and tetrazolyl compounds. However, in the case of pyrazolyl derivatives, the heterocyclic compound is also attached to the carbapenem nucleus at the C-4 position.

Other modifications of the structure introduced at the 2-position include a substituted vinyl group -C (R _a ) = CHR _b , in which, for example, R _a is hydrogen or methyl and R _b is hydrogen or lower alkyl (EP 0 330 108; Fujisawa) or are R _a and R _{b are} selected from hydrogen, 20 lower alkyl, aminocarbonyl, lower alkoxy, cyano, nitro and lower alkoxycarbonyl (EP 0 430 037, Banyu Pharmaceutical Co.). However, in the absence of the β-methyl substituent, such modification does not appear to confer stability on DHP-1.

PCT / GS 94/02347 discloses

Compounds of formula (C):

O • ···· 9 • · it

J ·· ··· · **

- 3 in which R stands for:

^ R ^and NN-V '.

r kde

R ^a represents a hydrogen atom, an optionally substituted (C 1-6) alkyl or an optionally substituted aryl;

R ^p represents a hydrogen atom, an optionally substituted (C 1-6) alkyl, or an alkyl or aryl group; R ^a and R ^e together form an optionally substituted 5 or 6 membered heterocyclic ring with or without additional heteroatoms;

R ^c is (C 1-8) alkyl, which is unsubstituted or substituted.

a fluorine atom or a hydroxyl group, which is optionally protected by an easily cleavable hydroxy protecting group, or?

an amino group which is optionally protected by an easily cleavable amino protecting group;

R ^d is hydrogen or methyl; and

-CO ₂ R ^e represents a carboxy group or a carboxylate anion or the group R ^{e is a} readily cleavable carboxyl protecting group.

SUMMARY OF THE INVENTION

According to the present invention there is provided compound (1):

Wherein R is selected from isobutyryloxymethyl, (5-methyl-2-oxo-1,3-dioxolen-4-yl) methyl and benzoyloxymethyl.

Particular compounds of the invention are: isobutyryl oxymethyl (5R, 6S) -2- [1-ethyl-5-methylpyrazol-S-yl] -e- (1R) -1-hydroxyethyl] -carbapen-2-em-3 -carboxylate, (5-methyl-2-oxo-1,3-dioxolen-4-yl) methyl (5R, 6S) -2- [1-ethyl-5-methylpyrazol-3-yl] -6 - [(1R)] -1-hydroxyethyl] -carbapen-2-em-3-carboxylate, and benzoyloxymethyl (5R, 6S) -2- [1-ethyl-5-methylpyrazol-3-yl] -6 - [(1R) - 1 hydroxyethyl] carbapen-2-em-3-carboxylate.

Since all of the carbapenem compounds of the present invention are intended for use in pharmaceutical compositions, it is clear that they must be in substantially pure form, for example at least 50%, more preferably at least 75% and preferably at least 95%. % purity (percentages are percent by weight).

Impure preparations of the compound can be used to produce cleaner forms used in pharmaceutical compositions. Whenever possible, preferably each compound of the present invention is obtained in crystalline form. If the compounds of the invention are allowed to crystallize or are recrystallized 20 from organic solvents, a crystallization solvent may be present in the crystalline product. These solvates also include the invention. Similarly, the compounds of the invention may be crystallized or recrystallized from water-containing solvents. In such cases, hydration water may be present in the crystalline product.

Stoichiometric hydrates are also within the scope of the present invention.

The carbapenem antibiotic compounds of the present invention may be present in compositions for administration by any suitable means for use in human or veterinary medicine, which compositions may be prepared by methods and procedures known in the art similar to other antibiotics and the invention therefore also encompasses pharmaceutical compositions containing the antibiotic compound. according to

- 5 -, -.

of the present invention together with a pharmaceutically acceptable carrier or vehicle. The compositions may be formulated for administration by any suitable route, such as oral, parenteral, or topical, although oral administration is a particularly preferred route. Compounds i

they may be in the form of tablets, capsules, powders, granules, lozenges, creams or liquid preparations, such as oral or sterile parenteral solutions or suspensions. Tablets and capsules for oral administration may be in unit dosage form and may contain conventional excipients such as binding agents, for example, acacia syrup, gelatin, sorbitol, tragacanth.

10-or polyvinylpyrrolidone-fillers such as lactose, sugar, corn starch, calcium phosphate, sorbitol or glycine; tabletting lubricants such as magnesium stearate, talc, polyethylene glycol or silica; disintegrants such as potato starch; or acceptable wetting agents, such as sodium lauryl sulfate. The tablets may be coated according to methods well known in conventional pharmaceutical practice. Oral liquid ...?

the compositions may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups, or elixirs, and may not be in the form of a dry product for reconstitution with water or other suitable vehicle before use. Such liquid compositions may contain conventional additives such as suspending agents, for example sorbitol, methylcellulose, glucose syrup, gelatin, hydroxyethylcellulose, carboxymethylcellulose, gel stearate or hydrogenated edible oils; emulsifying agents such as lecithin, sorbitan monooleate or acacia; non-aqueous vehicles (which may include edible oils), for example, almond oil, oily esters, glycerin, propylene glycol, or ethyl alcohol; preservatives such as methyl or propyl p-hydroxybenzoate or sorbic acid; and, if desired, conventional flavoring or coloring agents. Suppositories will contain conventional suppository bases such as cocoa butter or other glycerides.

For parenteral administration, liquid unit dosage forms will be prepared using the compound and a sterile vehicle, wherein:

I • t · · · ·

Water is preferred. The compound, depending on the vehicle and concentration used, can either be suspended or dissolved in the vehicle. In preparing solutions, the compound may be dissolved in water for injection and sterile filtered prior to filling into suitable ampoules or vials and capping. Advantageously, agents such as local anesthetics, preservatives and buffers may be dissolved in the vehicle. To enhance stability, the formulations may be frozen after filling into tubes and the water removed in vacuo. The dry lyophilized powder is then sealed in the vial and to reconstitute the liquid prior "~ 6 using the ^- One-vial-fed with-water-for-injection -----

Parenteral suspensions are prepared in substantially the same manner except that the compound is suspended in the vehicle instead of being dissolved and sterilization cannot be accomplished by filtration. The compound may be sterilized with ethylene oxide prior to suspension in a sterile vehicle. Preferably, a surfactant or wetting agent is added to achieve a uniform distribution of the compound in the composition.

The composition may contain from 0.1 to 99.5% by weight, preferably from 10 to 60% by weight of the active material, depending on the mode of administration. If the composition is in the form of dosage units, each unit will preferably contain 50 to 500 mg of the active ingredient. The dose for use in the treatment of an adult human will preferably be in the range of 100 mg to 12 g per day for an average adult patient (body weight of 70 kg), e.g.

Approximately 1.5 to 170 mg / kg per day. A suitable dose is from about 6 g per day.

The daily dose of the compound of the invention is suitably administered several times over 24 hours. Typically, 250 mg is administered four times a day and the dose and frequency most appropriate for each individual patient will vary depending on the age, weight, and response of the patient, and on various occasions the attending physician may choose a higher or lower dose and varying frequency of administration. . These dosing regimens are within the scope of the invention.

When the compounds of the invention are administered in the above 5 dosage range, there are no toxicological effects.

The present invention also encompasses a method of treating bacterial infections in humans and animals, the method comprising administering a therapeutically effective amount of an antibiotic compound of the present invention of formula (I).

In another aspect, the invention provides the use of a compound of formula (I) for the manufacture of a pharmaceutical composition for treating a bacterial infection.

The compounds of the present invention of formula (I) are effective against a broad spectrum of Gram-positive and Gram-negative bacteria and can be used to treat a wide range of bacterial infections 15, including infections in immunocompromised patients.

Among many other uses, the compounds of the invention are valuable in the treatment of infections of the skin, soft tissues, respiratory tract and urinary tract in humans and can also be used for the treatment of mastitis in cattle.

A particular advantage of the antibacterially active compounds of the present invention is their stability to β-lactamase enzymes and the associated activity against β-lactamase producing organisms.

The present invention further provides a process for the preparation of a compound of formula (I) wherein R is isobutyryloxymethyl or benzoyloxymethyl, the process comprising treating a compound of formula XCH ₂ OCOCHMe ₂ or XCH ₂ OCOCeH ₅ wherein X is a leaving group such as halogen, especially bromine or iodine. the corresponding compound of formula (I), R is an alkali metal cation.

AND

The reaction is typically carried out at a temperature between 0 and 60 ° C, for example at room temperature in an inert atmosphere such as argon and in a suitable organic solvent such as N-methylpyrrolidin-2-one. Other suitable solvent systems include N, N'5 dimethylformamide and N, N'-dimethylacetamide.

The present invention further provides a process for the preparation of compound (I) wherein R is (5-methyl-2-oxo-1,3-dioxolen-4-yl) methyl, comprising reacting a corresponding compound of formula (I) wherein R is an alkali cation metal, with a compound of the formula:

____________ _: Ma________________________

wherein X is a leaving group such as halogen, especially bromine 15 or iodine.

The reaction is typically carried out between 0 and 60 ° C, for example at room temperature, in a suitable organic solvent, for example: N-methylpyrrolidin-2-ane, and in the presence of an acid binding agent such as 2,6-lutidine. Other suitable acid binding agents are pyridine 20 and diisopropylethylamine. Other suitable solvents are N, N'-dimethylformamide and N, N'-dimethylacetamide.

Compounds of formula (I) wherein R is an alkali metal cation such as sodium are disclosed in International Patent Application No PCT / GB 94/02347 and in the following under Preparation 1.

The compound of formula ICH ₂ OCOCHMe ₂ (iodomethyl isobutyrate) and iodomethyl benzoate can be prepared from the corresponding chlorides by the Finkelstein reaction, which is well known to those skilled in the art.

Chloromethylizobutyrát chíoromethylbenzoát and may be prepared by esterifying isobutyric acid or benzoic * ·· · v »u · ·· · _Ie * ι

Chloromethyl chlorosulfate (Binderup et al., Synthetic Commun., 14 (9), 857-864 (1984)) The following examples further illustrate the present invention.

Examples of the invention

General Procedure: The solutions were dried using anhydrous magnesium sulfate and the solvents were removed by evaporation under reduced pressure using a rotary evaporator. Chromatography was performed on a silica gel 60 column with a particle size io <0.063 mm.

Example 1 lzobutyryloxymethyl- (5R, 6S) -2- (1-ethyl-5-methylpyrazol-3-yl) -6 - [(1R) ^-1-1 hydroxyethyllkarbapen-2-em-3-carboxylate

The product of Preparation 1 (200 mg, 0.61 mmol) was dissolved in Nmethylpyrrolidin-2-one (2 mL). The solution was below room temperature at room temperature;

a solution of isobutyryloxymethyl iodide (360 mg, 1.6 mmol) in N-methylpyrrolidin-2-one (0.5 mL) was added with nitrogen and after 0.5 h the reaction mixture was diluted with ethyl acetate (15 mL) and the solution was washed with water (3x15 ml),

2o 5% sodium thiosulfate solution (15 mL) and then saturated brine (15 mL). The organic extract was dried (Na ₂ SO 4) and concentrated to an oil which was chromatographed on silica gel eluting with an acetone / toluene gradient to give a pale yellow

The title compound was obtained as a pale yellow microneedle (100 mg, 40%), m.p. 107-108 ^e C; (Found: M ⁺ , 405.1899; MC ₂₀ H ₂₇ N ₃ O _e = 405.1900); ν _max (CHCl ₃ ) 3458, 3019, 1772 and 1734 cm ^-1 ;

8 _h (CDCl ₃ ) 1.19 (6H, d, J 7.1 Hz), 1.36 (3H, d, J 6.1 Hz), 1.40 (3H, t, J

7.2 Hz), 1.76 (1H, d, J 4.9 Hz), 2.29 (3H, s), 2.62 (1H, septet, J 7.1 * 9)

Hz), 3.18 (1H, dd, J 2.7, 6.5 Hz), 3.30 (1H, dd, J 9.0, 18.8 Hz), 3.64 (1H, dd, J 9.8, 18.8 Hz), 4.08 (2H, q, J 7.2 Hz), 4.16-4.30 (2H, d, J

5.6 Hz), 5.98 (1H, d, J 5.6 Hz) and 7.02 (1H, s).

Example 2 (5-methyl-2-oxo-1,3-diOXolen-4-yl) methyl (5R, 6S) - 6 - [(R) -1-hydroxyethyl] -2- (1-ethyl-5-methyl-pyrazole-3) -yl) carbapen-2-em-3-carboxylate

- ^; ----- Solution - (- 5R76S) -6 - [(- R -) - 1-Hydroxy-ethyl] -2 - (- 1-ethyl = 5 = methyllp.yr.a = —_______ i____ io zol -3-yl) carbapen-2-em-3-carboxylate (300 mg) in N-methylpyrrolidinone (4 ml) was mixed with 2,6-lutidine (about 30 mg) and then 4-bromomethyl-5-methyl was added -1,3-dioxol-2-one (290 mg) (F. Sakamoto, S. Ikeda and G. Tsukamoto, Chem. Pharm. Bull. 32, (6));

2241-2248 (1984) in tetrahydrofuran (1 mL). The mixture was stirred for 1.25 h. Ethyl acetate and water were added and the layers were separated. Vodná:

the layer was reextracted with ethyl acetate and the combined ethyl acetate layers were washed with water, saturated brine and then dried. (MgSO ₄ ). The filtered solution was stored in a freezer overnight and then evaporated and chromatographed on silica gel (230-400 mesh).

ASTM) (2.5 x 12 cm column), loaded with CH ₂ Cl ₂ / hexane, and eluted with ethyl acetate. Product containing fractions were combined and evaporated to give a colorless solid that was recrystallized from acetone to give (5-methyl-2-oxo-1,3-dioxolen-4-yl) methyl (5R, 6S) -6 - [(R) crystals -1-hydroxyethyl] 2 S 2- (1-ethyl-5-methyl-pyrazol-3-yl) carbapen-2-em-3-carboxylate, m.p. 148-151 ° C; v _max (CH ₂ Cl ₂ ) 3605, 2975, 1823, 1776, 1737 '(sh),'

1721, 1598, 1546, 1314, 1231 and 1182 cm ^-1 ; ν max (EtOH / nm 325 (s / dm ³ plague ¹ cm · ¹ 14,654), 260.5 (E / dm ³ plague ¹ cm -1 ¹³ 3404); 5 (CDCl 3) 1.36 (3H, d, J 6.2 Hz), 1.40 (3H, t, J ca. 7.3 Hz), 1.76 (1H, d, J 4.9 Hz),

2.21 (3H, s), 3.19 (1H, dd, J 2.7 & 6.5 Hz), 3.31 (1H, dd, J 9.0 & 18.7

Hz), 3.61 (1H, dd, J9.9 & 18.7 Hz), 4.08 (2H, q, J 7.3 Hz), 4.16-4.30 e · (2H, m) ), 5.00 & 5.07 (2H, ABq, J 13.9 Hz), 6.88 (1H, s) ppm; found (EI ms) m / z 417 (/ Vf).

Example 3

Benzyloxymethyl (5R, 6S) -2- (1-ethyl-5-methyl-pyrazol-3-yl) -6-r (1R) -1-hydroxyethylcarbapen-2-em-3-carboxylate

The product of Preparation 1 (200 mg, 0.61 mmol) was dissolved in N-methylpyrrolidin-2-one (2 mL). To the solution at room temperature under nitrogen for "solution-added" benzoyloxymethyljodidu- (336-mg of _r-1, 28-mmol), IO N-methylpyrrolidin-2-one (0.5 mL) and after 0.5 h the reaction The mixture was diluted with ethyl acetate (15 mL) and the solution was washed with water (3x15 mL), 5% sodium thiosulfate solution (15 mL) and then saturated brine (15 mL). The organic extract was dried (Na ₂ SO 4) and concentrated to an oil which was chromatographed on silica gel eluting with an acetone / toluene gradient to give a pale yellow solid which was recrystallized from diethyl ether to give the title compound as a white solid. (143 mg, 53%), mp 131-134 ° C; (Found: M ⁺ , 439.1743; M C22H2SN3O6 = 439.1743); ν max (CHCl 3) 3021, 1740, 1602, 1452 and 1440 20 cm ^-1 ; Δh (CDCl ₃ ) 1.34 (6H, m), 1.70 (1H, d, J 4.9 Hz), 2.25 (3H, s), 3.19 (1H, dd, J 2.4 6.3 Hz), 3.30 (1H, dd, J 8.8, 18.8 Hz), 3.64 (1H, dd, J 10.1, 18.8 Hz), 4.07 (2H , q, J 7.2 Hz), 4.18-4.31 (2H, m), 6.20 (2H, s), 7.42-7.48 (2H, M), 7.56-7 61 (1H, m) and 8.10 (2H, d, J 7.1 Hz) ppm.

.........,

»·» 4 4 • 4 4 • 4 4 B • 4 • 4 ·· · • 4 • * « • * 4 (B) ·· 4 · 44 ' 4 4

Preparation 1 Sodium (5R, 6S) -6-f (R) -1-hydroxyethyl-2- (1-ethyl-5-methylpyrazole-3-yl) carbapen-2-em-3-carboxylate (a) Ethyl 1-ethyl- 5-methyl-pyrazole-3-carboxylate

A solution of N-ethylhydrazinoxalate (12 g) in glacial acetic acid (100 mL) was cooled in an ice bath and ethyl 2,4-dioxovalerate (11.24 mL) was added. After the addition was complete, the mixture was stirred at room temperature and after about 45 min the mixture was warmed to dissolve the non-dissolved ethanoic hydrazinoxalate. The mixture was stirred for an additional 2 hours and then poured into about 300 mL of water / about 700 mL of ethyl acetate, and solid K ₂ CO ₃ was carefully added with stirring until neutral pH. After separation, the aqueous layer was extracted again with ethyl acetate. The combined ethyl acetate extracts were dried (MgSO ₄ ) and the solvents removed to give an oil. Chromatography on silica gel, eluting with CH ₂ Cl ₂ / hexane and eluting with a gradient of ethyl acetate / hexane (from 2: 8 to 1: 1), gave ethyl 1-ethyl-5-methylpyrazole-3-carboxylate as an oil (13, 2 g); _ν ™ χ ₍₂ 0Η α ₂₎ 1717, 1446, 1389 and 1219 cm @ ^-1; Δ (CDCl ₃ ) 1.38 (3H, t, J 7.2 Hz), 1.42 (3H, t, J 7.3 Hz), 2.30 (3H, s), 4.17 (2H, q, J: 7.3 20 Hz), 4.38 (2H, q, J 7.1 Hz), 6.55 (1H, s); (found m / z 182.1055 m / z C 9 H 14 N 2 O 2 = 182.1055).

(b) 1-Ethyl-5-methylpyrazole-3-carboxylic acid

Ethyl 1-ethyl-5-methylpyrazole-3-carboxylate (10.93 g) in ethanol with - A 3-2ηίοΗΉη2 and refluxed for 6 hours. Ethanol was removed on a rotary evaporator and ethyl acetate / water solution was added. The pH of the mixture was adjusted to 3.0 and the layers were separated. The aqueous layer was re-extracted with ethyl acetate. The combined ethyl acetate layers were extracted with excess aqueous NaHCO ₃ solution. NaHCO ₃ extract was poured into excess acid and the pH was then adjusted to 3 and NaCl was added to the solution. The mixture was then repeatedly extracted with ethyl acetate and the combined extracts were dried (MgSO ₄ ) and evaporated. The residue was triturated with diethyl ether to give the acid as a solid (5.65 g); v _max (CH ₂ Cl ₂ ) 2754, 2598, 1698, 1498, 1464, 1387 and

1233 cm ^-1 ; Δ (CDCl ₃ ) 1.40 (3H, t, J 7.3 Hz), 2.32 (3H, s) 4.19 (2H, q, J

7.3 Hz), 6.61 (1H, s) ppm; (Found m / z 154.0740. m / z C ₇ H ₁₀ N ₂ O ₂ = 154.0742).

(c) N-Methoxy-N-methyl-1-ethyl-5-methylpyrazole-3-carboxamide

1-Ethyl-5-methylpyrazole-3-carboxylic acid (5.25 g) in dry dichloromethane (100 mL) containing Ν, Ν-dimethylformamide (0.26 mL) was cooled in an ice bath and oxalyl chloride solution (3, 5 mL) was added. 27 ml) in dichloromethane (25 ml), added dropwise over 15 drops. The mixture was stirred in ice for 25 min and then allowed to warm to room temperature where gas evolution was observed. After 10 min the solvent was removed by evaporation in vacuo and toluene was added (2 x) to remove any residual HCl and oxalyl chloride present. The obtained acid chloride 20 was redissolved in dichloromethane and N, Odimethylhydroxylamine hydrochloride (3.61 g) was added. The mixture was cooled in an ice bath and pyridine (6.0 mL) was added. The mixture was then allowed to stir at room temperature for 1.5 hours and then diluted with water (100 mL) and washed with brine. The organic layer was then dried 25 (MgSO ₄ ) and evaporated to give an oil. This was again chromatographed on silica gel with ethyl acetate / hexanes to give the desired hydroxamate fractions (5.2 g) after evaporation as a solid; _{ν max} (CH ₂ Cl ₂ ) 2982, 2937, 1641, 1489, 1445, 1379 and 975 cm ^-1 ; Δ (CDCl ₃ ) 1.43 (3H, t, J 7.3 30 Hz), 2.29 (3H, s), 3.42 (3H, s), 3.76 (3H, s), 4.13 (2H, q, J 7.3 Hz), 6.49 (1H, s); (found m / z 197.1164. m / z C _{9 HI} N ₃ 0 ₂ = 197.1164).

(D) 3-acetyl-1-ethyl-5-methylpyrazole; (d) 3-acetyl-1-ethyl-5-methylpyrazole;

A solution of N-methoxy-N-methyl-5-methylpyrazole-3-carboxamide in dry tetrahydrofuran (60 mL) was cooled in an ice bath 5 and treated with a 3.0 M solution of methylmagnesium bromide in ether (11.08 mL). After stirring for 1.5 h, the mixture was poured into a mixture of methanol (100 mL) and 5 M aqueous HCl (10 mL) in an ice bath. The mixture was then evaporated to reduce the volume and mixed with a mixture of dichloromethane, water and saturated brine. After separation, the aqueous layer was 10; The combined dichloromethane extracts were dried (MgSO4) and evaporated to give an oil (2.26 g) which solidified on standing; _{ν max} (CH ₂ Cl ₂ ) 1680, 1446 1425, 1380, 1324, 1208 and 945 cm ^-1 ; Δ (CDCl ₃ ) 1.44 (3H, t, J 7.3 Hz), 2.30 (3H, s), 2.53 (3H, s), 4.13 (2H, q, J 7.3) Hz), 6.51 (1H, s);

(Found m / z 152.0949. m / z C _and H ₁₂ N ₂ O - 152.090).

(e) (3S, 4R) -4-f (1-Ethyl-5-methyl-piperazin-3-yl) -carbonylmethyl-3 - [(R) -1- tert -butyldimethylsiloxylmethyl-azetidin-2-one

A solution of 3-acetyl-1-ethyl-5-methylpyrazole (3.51 g) in dry

2-tetrahydrofuran (THF, 150 mL) was cooled under argon in an acetone / solid carbon dioxide bath, and 1 M lithium bis (trimethylsilyl) amide solution (50 mL) was added. The mixture was stirred for 45 min and then (3R, 4R) -4-acetoxy-3 - [(1R) -1-tert-butyldimethylsilyloxyethyl] azetidinone (6.6 g) was added as a solid under argon. The mixture was

25__v. Then, saturated aqueous ammonium chloride solution and ethyl acetate were added and the mixture was allowed to warm to room temperature. A small amount of water was added and the layers were separated and the aqueous layer was re-extracted with ethyl acetate. The combined ethyl acetate extracts were washed with saturated brine, dried and evaporated. Silica gel chromatography eluting with solution

ethyl acetate / hexanes gave the title compound (3.65 g), in _max (CH ₂ Cl ₂ ) 3411, 1761, 1678, 1376, 1151 and 838 cm ^-1 ; Δ (CDCl ₃ ) 0.064 (6H, s), 0.86 (9H, s), 1.20 (3H, d, J 6.3 Hz), 1.44 (3H, t, J 7.3 Hz) 2.31 (3H, s), 2.89 (1H, dd, J 1.8 & 4.9 Hz), 3.15 (1H, dd, J 10.0 & 17.1 Hz), s 3 50 (1H, dd, J 3.5 & 17.0 Hz), 4.06-4.25 (4H, m), 6.11 (1H, s), 6.53 (1H, s); (found m / z 379.2296, m / z C 19 H 33 N 3 O 3 Si = 379.2291).

(f) Allyl (2Ra2S) -2 - {(3S, 4R) -4 - [(1-ethyl-5-methylprazole-3-yl) carbonylmethyl-3-f (R) -1-tert-butyldimethylsilyl] oxyethyl-2-oxoazetidin-1-yl -2-hydroxyacetate -

(3S, 4R) -4 - [(1-Ethyl-5-methylpyrazol-3-yl) carbonylmethyl] -3 - [(R) -1-tert-butyldimethylsilyloxyethyl] azetidin-2-one solution (3.6 g) and allyl glyoxylate hydrate (1.66 g) in toluene (100 mL) was heated under reflux in a Dean-Stark apparatus under argon atmosphere for 15 hours for 3.5 hours. Thin layer chromatography of the reaction mixture showed near completion of the reaction, so 190 mg of allyl glyoxalate hydrate and the mixture was refluxed for an additional 45 min. The mixture was cooled, toluene was removed to give crude allyl (2R and 2S) -2 - {(3S, 4R) -4 - [(1-ethyl-5-methylpyrazol-3-yl) carbonyl-20-methyl] -3- [ (R) -1-tert-Butyldimethylsilyloxyethyl] -2-oxoazetidinyl} -2-hydroxyacetate which was used in the next step; _νmax (CH ₂ Cl ₂ ) 3681, 3518, 1758, 1676, 1448, 1376, 1326, 1209, 1148, 1092, 954 and 836 cm ^-1 ; Δ (CDCl ₃ ) m / z 0.035 (s), 0.061 (s) (together 6H). 0.858 (s), 0.865 (s) (together 9H), 1.21 (d, J 6.2 Hz), 1.34 (d, J 6.2 Hz), 25 (together 3H), 1.44 ( 3H, t, J 7.2 Hz), 2.31 (3H, s) 2.95-3.00 (1H, m), 3? 25 ^ 3 ^ Γ (Μ ^ η) Γ6753 ”(3) 76756 (sjrppm: —---------- ··

in*

(q) Allyl 2 - {(3S, 4R) -4 - [(1-ethyl-5-methylprazrazol-3-yl) carbonylmethyl] -3-I [(Ρ) -1-Γ6Γο-όυίγΙάϊΓηΒΐΚνΐ5ίΙνΙοχγ6] } -2- (tert-Butylphosphoranylidene) acetate

Allyl (2R and 2S) -2 - {(3S, 4R) -4 - [(1-ethyl-5-methylpyrazol-3-yl) carbonyimethyl] -3 - [(R) -1-tert-butyldimethylsilyloxyethyl] solution -2-oxoazetidinyl} -2-hydroxyacetate (crude from above) in dry THF (125 mL) under argon was cooled to

- 20 ° C and mixed with 2,6-lutidine (1.98 mL), then thionyl chloride (1.24 mL) was added. The mixture was stirred for 30 min at - 20 ° C and then IB pqnecíTáňšrofířáTna ^_ pOkojovouteplotu-A-wherein ---------- filtered residue was washed with THF (20 mL). The filtrate was evaporated in vacuo, 70 mL toluene was added and again removed in vacuo, and the residual oil was dried in vacuo. The oil was then taken up in 1,4-dioxane (40 mL) under argon and treated with tri-n-butyiphosphine (3.11 mL).

The mixture was stirred for 1 h. 2,6-lutidine (1.59 mL) was then added and the mixture was stirred for an additional 30 min. The mixture was diluted with ethyl acetate, washed with water and brine and dried (MgSO ₄ ). After removal of ethyl acetate, the crude product was chromatographed on silica gel eluting with ethyl acetate / hexanes to give

The phosphorane used in the next step.

(h) Allyl 2 - ((3S, 4R) -4-f (1-ethyl-5-methylprazrazol-3-yl) carbonylmethyl-1-3. [(R) -1-hydroxyethyl-2-oxoazetidin-1-yl} -2- ( tri-n-butylphosphoranylidene) acetate

The phosphorane prepared above was taken up in 1,4-dioxane (60 ml) and 20 mL of 5M HCl was added. After 1 hour, approximately 40 mL of saturated aqueous NaHCO ₃ solution was carefully added to the mixture and then solid.

NaHCO _{3 l} has been obtained until slightly basic pH. Saturated brine was added and the mixture was extracted twice with ethyl acetate.

3o. The combined extracts were dried over MgSO ₄ and evaporated. The residue was chromatographed on silica gel eluting with ethyl acetate / hexane to give i

providing a hydroxy compound (2.60 g), v _max (CH ₂ Cl ₂₎ 3454, 1741, 1667, 1606, 1448; 1403, 1379, 1155, 1087, 953, and 811 ^{cm-1 *.}

(i) Alvi (5R, 6S) -6f (R) -1-hydroxyethyl-241-ethyl-5-methylprazrazol-3syl) carbapen-2-em-3-carboxylate

Allyl 2 - {(3S, 4R) -4 - [(1-Ethyl-5-methylpyrazol-5-ylcarbonylmethyl) -3 - [(R) -1-hydroxyethyl] -2-oxoazetidinyl} -2- (tri-) solution n-butylphosphoranylidene) acetate (2.6 g) in toluene (120 ml) containing hydroquinone

---- (20-ma) -blue under reflux in an argon atmosphere for 4 hours, allowed to stand for 64 hours and then heated under reflux for a further 2 hours. The mixture was cooled and then applied to the column. (4.5 x 12 cm) silica gel (particle size 0.040-0.063 mm) eluting with ethyl acetate / hexane; 1: 1; 6: 4; 7: 3; 8: 2; 9: 1 (250 ml each) and then ethyl acetate. This gave 463 mg of carbapenem;

_{ν max} (CH ₂ Cl ₂ ) 3604, 2976, 1774, 1716, 1600, 1546, 1311, 1189 cm ^-1 ; Λ max (EtOH) / δ 321.5 (ε / dm ³ mor ¹ cm ^-1 14.856), δ (CDCl 3) 1.36 (d, J 6.3).

Hz), 1.39 (t, J 7.3 Hz) (together 5H), 1.80 (1H, d, J 5.0 Hz), 2.28 (3H,

s), 3.19 (1H, dd J 2.7 & 6.7 Hz), 3.28 (1H, dd, J 9.0 & 18.6 Hz), 3.60 (1H, dd, J 9 9 & 18.5 Hz), 4.08 (2H, q, J 7.3 Hz), 4.16-4.30 (2H, m),

4.68-4.90 (2H, m), 5.27 (1H, m, ca. d, J ca. 12 Hz), 5.46 (m, ca.

d, J ca. 17 Hz), 5.93-6.08 (1H, m), 7.00 (1Ή, s) ppm; (found m / z 345.1693. m / z C 18 H 23 N 3 O 4 = 345.1689).

(i) Sodium salt (SRSeíííRj-1-hydroxethylene-Z-d-ethyl-5-methyl-2'-pyrazol-3-yl) -kg-baoene-2-em-3-carboxylate ______ .__________

A solution of allyl (5R, 6S) -6 - [(R) -1-hydroxyethyl] -2- (1-ethyl-5-methylpyrazol-3-yl) carbapen-2-em-3-carboxylate (267 mg) in dichloromethane ( 3 ml) and ethyl acetate (3 ml) under argon were mixed with sodium 2-ethylhexanoate (183 mg), triphenylphosphine (24 mg) and then 30 tetrakis (triphenylphosphine) palladium (0) (35 mg) and the mixture was stirred for 45 min.

The mixture is stirred. 100 ml of diethyl ether was then added and after 90 min the mixture was centrifuged. The residual solid was dried in a stream of argon and then in a desiccator. The solid was then taken up in sodium chloride water and chromatographed on 5 DIAION HP20SS resin eluting with water and water / THF; 1%, 2% and 3%

THF. Fractions were monitored by HPLC and those containing the product were combined, concentrated, and lyophilized to give (5R, 6S) -6 [(R) -1-hydroxyethyl] -2- (1-ethyl-5-methylpyrazole) sodium salt. 3-yl) carbapen-2-em-3-carboxylate as a solid (168 mg); v _max (KBr) 1761, io — 1608-1-57-7-138-1 ^ -1-2-25-cm ^: ^ X _m and (.H7.Q) / nm_298_ (s / dm ³ plague ¹⁾ cm ^-1

8,531); Δ (D ₂ O) 1.26 (d, J ca. 6 Hz); 1.27 (d, J ca. 7 Hz) (together 5H), 2.23 (3H, s), 3.17 (2H, approx. D, J ca. 9 Hz), 3.44 (1H, dd) J (2.9 & 6.0 Hz), 4.04 (2H, q, J 7.3 Hz), 4.15-4.25 (2H, m), 6.41 (1H, s) ppm.

Claims (5)

PATENT CLAIMS ·. ω wherein R is selected from isobutyryloxymethyl, (5-methyl-2 oxo-1,3-dioxolen-4-yl) methyl and benzoyloxymethyl. A compound which is isobutyryloxymethyl (5R, 6S) -2- [1-ethyl-5-methylpyrazol-3-yl] -6 - [(1R) -1-hydroxyethyl] carbapen-2-em-3- 15 carboxylate. 3, The compound is (5-methyl-2-oxo-1,3-dioxolen-4-yl) methyl (5R, 6S) -2- (1-ethyl-5-methylpyrazol-3-yl) 6 - [(1 R) -1 hydroxyethyl] carbapen-2-em-3-carboxylate. 4. A compound which is (5R, 6S) -2- [1-ethyl-5-methylpyrazol-3-yl] -benzoyloxymethyl-1,1-carbaphen-2-em- 3 carboxylate. 5. A pharmaceutical composition comprising a compound according to any one of the preceding claims. 8. 8. 9. 9. of the claims together with a pharmaceutically acceptable carrier or vehicle. A method of treating bacterial infections in humans and animals, comprising administering a therapeutically effective amount of a compound according to any one of claims 1 to 4. Use of a compound according to any one of claims 1 to 4 for the manufacture of a pharmaceutical composition for the treatment of a bacterial infection. A process for producing a compound of formula (I) according to claim 1, wherein R is isobutyryloxymethyl or. benzoyloxymethyl, comprising reacting a corresponding compound of formula (I) wherein. R is an alkali metal cation, with a compound of formula XCH ₂ COCCHMe ₂ or XCHiOCOC ₆ H ₈ , wherein X is a leaving group. A process for the preparation of a compound of formula (I) according to claim 1, wherein R is (5-methyl-2-oxo-1,3-dioxolen-4-yl) methyl, comprising reacting a corresponding compound of formula (I), wherein R is an alkali metal cation, with a compound of the formula wherein X is a leaving group