FI80040B - FOERFARANDE FOER FRAMSTAELLNING AV OPTISKT AKTIVA PENEMER. - Google Patents

Description

80040 METHOD FOR THE PREPARATION OF OPTICALLY ACTIVE (5R) -2-CARBAMOYL OXIMETHYL-LI-6 (S) - [1 (R) -HYDROXYETHYL] -2-PENEM-3-CARBOXYLIC ACID - OPTFRIVAMIS-FÖRFARANDE CARBAMOYLXXIMETHYL-6 (S) - [1 (R) -HYDROXYETHYL] -2-PE-NEM-3-CARBOXYLSYRA - DIVIDED FROM APPLICATION 824192 -AVDELAD FRÄN For the preparation of 5R) -2-carbamoyloxymethyl-6 (S) - [1 (R) -hydroxyethyl] -2-penem-3-carboxylic acid or a pharmaceutically acceptable salt thereof

OH

f 8 ^ ^ (I)

g ^^ COOH

The compound of formula (I) is prepared according to the invention according to the following reactions. In the formulas, Ph represents a phenyl group, and the meanings of the groups PG, R and R 1 are given below.

- COOR

2% nm

COOR

UV) (IVa) * / I ’

(V) COOR

2 80040

If ope - »4 (f fHCl - *

COOR

CXI) Ά ffc n r — v · *, r ~ Nv ^ 3 (m) COOR (xiii) C00R coor

OR. I OR- I OH I

4-t ^ · '' ^ ΖΓΧ "Jj = DT" · i - »- ^ COOR / </” “* 6 (xv) (m) (I)

The process according to the invention is thus characterized in that the compound of the general formula II .....

COOR

wherein R is p-nitrobenzyloxycarbonyl, dimethyl-1ert. -butylsilyl, diphenyl-tert-butylsilyl, 2,2,2-trichloroethoxycarbonyl, trimethylsilyl, benzyl, p-bromophenacyl, triphenylmethyl or pyranyl and R is lower alkyl, 2,2,2-trichloroethyl , acetonyl, allyl, benzyl, p-nitrobenzyl, p-methoxybenzyl, phenyl, o-nitrophenyl, benzhydryl or 1-phenoxyethyl, the hydroxy group is protected by a protecting group PG which is e.g. p-nitrobenzyloxycarbonyl, dimethyl-tert-butyl difenyy1i-in tert -butyl

II

3,80040 lisilyl, 2,2,2-trichloroethoxycarbonyl, trimethylsilyl, benzyl, p-bromophenyl, triphenylmethyl or pyranyl to give a compound of formula III

Jj Ϊ

Tcr- -

COOR

wherein, R, R 1 and PG are as defined above, the phosphorus tribromide 11a is reduced to give the compound of formula IV OR,> r · .....

wherein R, R 1 and PG are as defined above, are ozonolysed in a solvent at -80 to -50 ° C to give the compound of formula IVa OR 1

Qf

COOR

wherein R, R and PG are as defined above, are methanolized in the presence of silica gel to give a compound of formula V ORx -χτ * '4 80040 wherein R1 and PG are as defined above, condensed at 70-100 ° C to give a compound of formula CHOCO3R with a compound in which R is as defined above, that the compound of formula X obtained

FLH

ö (X) / ~ y

COOR

wherein R, R 1 and PG are as defined above, chlorinating with thionyl chloride to give the compound of formula X1 obtained is P 2 - [>, S-V 2, XopS, (X.)

COOR

wherein R, R and PG are as defined above, are reacted with triphenylphosphine at a temperature of 30 to 60 ° C, and that the obtained compound of formula XII

JN

, (XII) ^ -λνγ ».ΡΡΙι3

° COOR

wherein R, R and PG are as defined above, are converted to a compound of formula I by carrying out the following reaction steps (a) to (c) in the order shown or by performing step (a) after step (b) but before step (c) , or so that step (a) is performed after steps (b) and (c), or when the group is hydrogen, step (b) is omitted and step (a) is performed before or after step (c): (a) ring formation is performed by heating under a nitrogen atmosphere in an inert solvent at a temperature in the range of 80-150 ° C, 5 80040 (b) deprotecting the substituent of the following formula with trifluoroacetic acid in ethyl acetate or tetrabutylammonium fluoride in tetrahydrofuran, optionally in the presence of acetic acid, to the next substituent of the formula by reaction with trichloroacetyl isocyanate in acetone and treatment with silica gel in methanol, and finally deprotection to give a compound of formula I which, if desired, is converted into a salt.

EP-A1 0 013 067 discloses a process for the preparation of 6-unsubstituted penems in which the hydroxyethyl group has a hydrogen at the 6-position.

The method according to the invention differs from the formula

ψ H

______f l'5 VS ^ 'CH2OPG

o ° to obtain an intermediate completely from said known method.

In fact, said publication describes the reaction which takes place between a compound of formula HS-C-CH 2 -OSi 2 BDP and a compound of formula O-OAc

J-NH

0 6 80040 (cf. Example 4).

Coupling of a group of formula -S -C - CH2 - OSi ^ BDP Il 2

O

in this way, the 4-position of the azetidinone ring leads to the formation of a racemic compound having the 4 R, S configuration.

In contrast to the above, the process according to the invention, which is based on an optically active compound of the formula, makes it possible to prepare optically active 4R-azetidinone, which is a completely unexpected and at the same time advantageous result.

The invention provides a process for the preparation of compound (X) and three different ways of preparing compound (I) from compound (XII). All of these have a common reaction step in which compound (X) is converted to compound (XII).

In the preparation of compound (X), the free hydroxy group of compound (II) (prepared according to British Patent No. 2,043,639) is protected with a protecting group, the sulfoxide function of the obtained compound (III) is reduced with phosphorus tribromide, both carbon-carbon double bonds of the obtained compound (IV) are ozonolized in a solvent at a temperature of 80 to -50 ° C, the N-substituent of the obtained compound (IVa) is methanolized and the obtained compound (V) is condensed with a glyoxylic acid ester of the general formula CHOCO 3 R, wherein R is as defined above.

The reduction can be performed using phosphorus tribromide in a temperature range of -40 to -20 ° C in a solvent such as anhydrous dimethylformamide. The ozonolysis can be carried out at a temperature in the range from -80 to -50 ° C in a solvent such as diethyl ether, methanol or especially dichloromethane. The methanolysis is preferably carried out in the presence of silica gel or using a catalytic amount of a strong base such as sodium methoxide. Condensation of compound (V) with glyoxylic acid ester is conveniently carried out at an elevated temperature, such as 70-100 ° C, in an organic

In a solvent such as benzene or toluene.

11 7 80040

A common reaction step for converting compound (X) to compound (XII) involves chlorination of compound (X) and reaction of compound (XI) thus obtained with triphenylphosphine. The chlorination is preferably carried out at thionyl chloride 11a at a temperature in the range of -20 to 0 ° C in an inert solvent such as tetrahydrofuran. The reaction with triphenylphosphine may be carried out at 30 to 60 ° C, preferably 40 ° C, in an organic solvent such as tetrahydrofuran in the presence of a base such as pyridine or lutidine. On the other hand, it can be performed in the presence of silica gel at ambient temperature in a shorter time.

All three methods of preparing compound (XII) from compound (I) involve three steps: ring formation, removal of the protecting group PG, and attachment of the desired group CONH2. The attachment of the CONH 2 group, of course, takes place only after the deprotection, and the methods of preparation differ only in whether the ring formation is carried out in the first, second or third of these three steps. The ring formation is carried out by heating under a nitrogen atmosphere at a temperature in the range of 80 to 150 ° C in an inert Liu solvent such as benzene, toluene or xylene. The removal conditions of the protecting group PG depend on the nature of the protecting group PG. The CONH2 group can be attached by reaction with an isocyanate such as trichloroacetyl isocyanate or chlorosulfonyl isocyanate.

The two features of this invention require further consideration. First, the carbon atom at position 5, whose R-configuration "is the only essential stereochemical requirement for antibiotic activity" (HR Pfaendler, J. Costeli and RB Woodward, J. Am. Chem. Soc. 1.01, 1979, 6306), retains its configuration from compound (II ) to compound (I). The Hii1i sulfur bond is not destroyed at any stage. Second, the incorporation of the CONH2 group occurs at a very late stage in the synthesis.

e 80040

The compound of the invention has antibacterial activity and β-lactamase inhibitory activity, for which it is more effective than the compounds known from GB patent 2,043,639. This is apparent from the following table, which compares the two compounds disclosed in said publication with the compound of the invention.

MIC pg / ml PCE

PCE 22101

Bacterial strain 20077 / B40 / 341 Compound A

Staphilococcus aureus - 209 P 0.39 0.39 <0.018 - Smith 0.19 0.39 0.018

Escherichia coli B 1.S6 0.78 0.39 B cef R> 100> 50 0.78

Bnterobacter

Cloacoe P 99> 50> 50 0.78

Klebsiella aerogenes 1082 E ^ 50 ^ 50 0.78 pneumoniae ATCC 10031 - 3.12 0.39

In table FCE 20077 / B40 / 341 is (5R) -acetoxymethyl-2-acetoxymethyl-2-penem-3-carboxyclyl-111i according to GB publication 2,043,639, compound A is (5R) -α-cetoxymethyl-2 according to GB publication - (1-methyl) -1H-tetrazol-5-yl) -thiomethyl-2-penem-3-carboxylate, and FCE 22101 is sodium (5R, 6R) -2-carbamoyloxymethyl-6 - [(1R) -hydroksie-tyylij-2-penem-3-carboxylate.

The invention is illustrated by the following examples in which the abbreviation PNB means p-nitrobenzyl, TBDPS tert-butyldiphenylsilyl and TBDMS tert-butyldimethylsilyl.

II

9 80040

Example i: 4 (R) -tert-Butyldiphenylsilyloxyacetylthio-3 (S) - {λ (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-hydroxymethyl) -azetidine-2- one

Qf02PKB OCOjPNB

- ^^ Xj ^^^ OTBDPS - | ^ S ^ s | ^^ Vt) TBDPS

o) - ° J-ΝΎ ° Η

COOPITB

A solution of 3.11 g (5 mmol) of 4 (R) -tert-butyldiphenylsilyloxyacetylthio-3 (S) - (λ (R) -p-nitrobenzyloxycarbonyloxyethyl) azetidin-2-one and 3.20 g (12, 5 mmol) of p-nitrobenzylglyoxylate in 100 ml of benzene, was refluxed so that water was removed by azeotropic distillation (5 ml) to near dryness.

10 80040

Example 2: 4 (R) -tert-butyldiphenylsilyloxyacetylthio-3 (S) - [λ (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-chloromethyl) azetidine- 2-one oco2? Nb oco2pnb

- | ^ S'V / ^ SÖTBDPS -f '^^^ OTEDPS

J · γΗ4 - Y— "γοϊ

COCPNB

COOPNB

A stirred solution of 3.5 g (4.2 mmol) of 4 (R) -tert-butyldiphenylsilyloxyacetylthio-3- (S) - [T- (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl) -1-hydroxymethyl) -azetidin-2-one in dry tetrahydrofuran at 0-5 ° C, treated with 0.48 ml (6 mmol) of pyridine and 0.43 ml (6 mmol) of thionyl chloride. After half an hour the reaction mixture was filtered and the filtrate was evaporated to dryness insurance, where, to give the chloro ester as a yellow gum.

Example 3: 4- (R) -tert-Butyldiphenylsilyloxyacetylthio-3 (S) - [T (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl) azetidine -2-one

OCO-PNB

f 2 „oco2pnb

Kr ~~ - γτ- COOPHB ° J 3

COOPNB

A solution according to the previous example of 4 (R) -111180040 tert-butyldiphenylsilyloxyacetylthio-3 (S) -H (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-chloromethyl) -azetidine- 2-one in tetrahydrofuran, was treated with 2.2 g (8.5 mmol) of Ph 2 P and 20 g of silica gel. The mixture was evaporated to dryness in vacuo and the resulting powder was left at room temperature for 2 hours. The powder was then applied to the top of the column and the phosphorane was eluted with cyclohexane-ethyl acetate mixtures to give 3.2 g of the title compound as a pale yellow foam.

Example 4; 4 (R) -Hydroxyacetylthio-3 (S) -R (R) -p-nitrobenzyloxycarbonyloxyethyl-1- (1-p-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl) -azetidin-2-one

OCO.PNB

i 2 oco2pnb <r γ ^ 3 ^ * \ ^ pph ^

COOPNB COOPNB

4 ml of trifluoroacetic acid was added to a stirred solution of 1.07 g (1 mmol) of 4 (R) -tert-butyl-diphenylsilyloxyacetylthio-3- (S) - β (R) -p-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl ) -azetidin-2-one in 50 ml of ethyl acetate. After 15 minutes, the solvent was removed, 50 ml of toluene was added and the solvent was evaporated again to give 1.3 g of the phosphonium salt, which was dissolved in 50 ml of tetrahydrofuran and treated with 4 equivalents of tetrabutylammonium fluoride (TBAF).

After 1 hour, the mixture was evaporated to dryness and dissolved in 50 ml of ethyl acetate and washed first three times with 25 ml of saturated sodium hydrogen carbonate solution and then with 25 ml of water.

The organic phase was separated, dried over anhydrous sodium sulfate and evaporated to dryness in vacuo.

Chromatography of the oily residue on silica gel (cyclohexane / ethyl acetate) gave 0.75 g of the title compound as a foam.

Example 5: p-Nitrobenzyl 5 (R) -2-hydroxymethyl-6 (S) -R (R) -p-nitrobenzyloxycarbonyloxyethyl] -2-penem-3-carboxylate

OCO ^ PNB 0CO2PNB

ϋ γ 3 COOPNB

COOPKB

A solution of 0.6 g of 4 (R) -hydroxyacetylthio-3 (S) - / T (R) -p-nitrobenzyloxycarbonyloxyethyl? ” 1- (1-p-Nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl) -azetidin-2-one in 200 ml of toluene was refluxed for 2 hours with a catalytic amount of hydroquinone. The solvent was then evaporated off in vacuo and the residue was purified by column chromatography on silica gel using toluene-ethyl acetate mixtures as eluent to give 0.42 g of the title compound.

UV:> (ΔtOH 95%) 260 nm (¢: .9100), 319 nm (<£ 8400)

Π * 3 X

IR: ν (CHCl 3, J 3500-3200, 1790, 1745, 1710, 1605, 1580 cm -1 PMR (CDCl 3): 1.51 (3H, d, J = Hz), 3.99 (H, dd , J = 2.7.5Hz), 4.69 (2H, broad s), 5.15 (1H, m), 5.23 and 5.46 (2H, centers of ABq, J = 14Hz), δ , 26 (2H, s), 5.64 (1H, d, J = 2Hz), 7.51 (2H, d, J = 8Hz), 7.61 (2H, d, J = 8Hz), 8.20 (4H, d, J = 8Hz).

i3 80040

Example 6: p-Nitrobenzyl (5R) -2-tert-butyldiphenylsilyloxymethyl-6 (S) - [T- (R) -p-nitrobenzyloxycarbonyloxyethyl] -2-penem-3-carboxylate

OCO ^ PNB 0C0? PNB

^ ^ Y ^^ OTBDPS H- -fSNVx ^> vbrBDPS

j NYpph * / -M \

0 j 3 o COOPNB

COOPNB

A solution of 0.3 g of 4 (R) -tert-butyldiphenylsilyloxyacetylthio-3 (S) - [T (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1- triphenylphosphoranylidenemethyl) -azetidin-2-one in dry toluene, refluxed for 3 hours. The solvent was removed and the mixture was chromatographed on silica gel using cyclohexane-ethyl acetate mixtures as eluent to give 0.12 g of the title compound.

Example 7: p-Nitrobenzyl (5R) -2-hydroxymethyl-6 (S) - [T (R) -p-nitrobenzyloxycarbonyloxyethyl] -2-penem-3-carboxylate qco2pnb oco2pnb

0 COOPNB COOPNB

A solution of 0.1 g of p-nitrobenzyl (5R) -2-tert-butyldiphenylsilyloxymethyl-6 (S) - [T (R) -14 B0040 p-nitrobenzyloxycarbonyloxyethyl] -2-penem-3-carboxylate in tetrahydrofuran, treated with 3 equivalents of TBAF at -15 ° C with stirring. The reaction mixture was then poured into 50 ml of ethyl acetate and washed three times with 30 ml of water. The dried organic phase was evaporated to dryness and chromatographed on silica gel using ethyl acetate-cyclohexane mixtures as eluent to give 20 mg of the title compound.

This material proved to be identical to that obtained in Example 5.

Example 8: Sodium (5R) -2-hydroxymethyl-6 (S) -1T (R) -hydroxyethyl] -2-penem-3-carboxylate oco2pnb ^

- ^ SVvj OH

0 ^ “N ^ COOPNB 0 ^ N ^ OONa

To a solution of 54 mg of p-nitrobenzyl (5R) -2-hydroxymethyl-6 (S) - [T (R) -p-nitrobenzyloxycarbonyloxyethyl] -2-penem-3-carboxylate in a mixture of ethyl acetate and water containing 6 mg of sodium hydrogen carbonate, 40 mg of 5% Pd / C was added. The mixture was hydrogenated for 1 h at ambient pressure. An additional 20 mg of 5% Pd / C was added and allowed to stand for 0.5 h. The mixture was filtered and the aqueous phase was separated and washed with ethyl acetate.

After evaporation of the aqueous phase to dryness, the residue was purified on a reflux column (Umkehrphasunsaule) using water as eluent. There was thus obtained 12 mg of the title compound as an amorphous solid.

UV: λ (EtOH 95%) 263 nm, 304 nm.

max is 80040

Example 9: 4 (R) - (1-tert-Butyldimethylsilyloxymethyl-vinylthio) -3 (S) -β (R) -p-nitrobenzyloxycarbonyloxyethyl-1- (1-methoxycarbonyl-2-methyl-1-propenyl) azetidin-2-one-S-oxide

0C0 PNB o QC 0 PNB O

X? f3 -3 M T 0,1_ „i ΓΤί“ 3 0> —0J— <i ch3 3

COOCH

3 COOCH

1.9 g of 4 (R) - (1-hydroxymethylvinylthio) -3 (S) - β (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-methoxycarbonyl-2-methyl-1-propenyl) azetidin-2-one oxide was dissolved in 20 ml of dichloromethane.

0.7 ml of triethylamine, 640 mg of tert-butyldimethyldiyl chloride and 20 mg of dimethylaminopyridine were added under a nitrogen atmosphere. The solution was stirred overnight at room temperature, washed with water and ammonium chloride solution and the solvent was evaporated. The residue was chromatographed on silica gel using cyclohexane-ethyl acetate (1: 1) as eluent to give 0.83 g of the title compound.

δ 80040 PMR (CDCl 3): δ (ppm) 0.07 (s, 6H, SIFCH 2), 0.88 (s, 9H,

SiC (CH) n), 1.41 (d, J = 6.5 Hz, 3H, CH CH), 2.14 (s, 3H, = ^ - 3),

2.30 (s, 3H, β-3), 3.75 (s, 3H, COOCH 3), 3.7-3.8 (m, 1H, H-6), 4.48 (broad s, 2H , CH OSi), 5.25 (s, 2H, CH 2 Ph), 5.1-5.2 (m,? H, H-5, CH 3 CH), 5.85 (broad s, 1H, -), 5.98 (broad s, 1H, = ^ -), 7.4-8.4 (m, 4H, PhNO 2), IR (CH-Cl 2): δ (cm -1): 1730 C = O unsaturated ester , 1755 C-0 C. £.

OCOO, 1780 C = O / 3-lactam.

Mass spectrum (FC): m / e 624.

Example 10: 4 (R) - (1-tert-Butyldimethylsilyloxymethyl-vinylthio) -3 (S) - [1 (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-methoxycarbonyl-2-methyl-1- propenyl) azetidin-2-one

OCO ^ PNB 0 OC0 PNB

1 “h l [TBDMS f H _ J -> <” <, * "> = <

COOCH „COOCH

3 3 TBDMS means tert-butyldimethylsilyl

A solution of 0.8 g of 4 (R) - (1-tert-butyldimethylsilyloxymethylvinylthio) -3 (S) - [T (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- {1-methoxycarbonyl-2 -methyl-1-propenyl) -azetidin-2-one S-oxide in 30 ml of anhydrous dimethylformamide, cooled to -20 ° C and 0.25 ml of phosphorus tribromide was added.

After 15 minutes, the mixture was diluted with ethyl acetate, washed twice with saturated sodium hydrogen carbonate solution and then with water, and dried over sodium sulfate. Evaporation of the solvent gave 0.7 g of the title compound.

PMR (CDCl 3) δ (ppm): 0.05 (s, 6H, Si (CH 2) p), 0.90 (s, 9H, 3 e e.

SiC (2'12) 3) '1.48 (d' J = 6'5Hz · £ H3CH). 2 · 01 (s · 3H · 3> - 2.24 (s, 3H, = ^ - 3), 3.35 (dd, J = 2.5, 7, = Hz, 1H, H-6), 3.73 (s, 3H, C 20 CH 3), 4.08 (t = J = 2.0 Hz, 2H, CH 2 OH), 5.26 (s, 2H, CH 2 Ph), 5.2-5.35 (m, 3H CH_CH, H-5, = <-), 5.56 (d, J = 2.0, 1H, --6 = ^ -), 7.4-8.4 (m, 4 H, PhNo).

Example li: 4 (R) -tert-Butyldimethylsilyloxyacetylthio-3 (S) - [(R) -p-nitrobenzyloxycarbonyloxyethyl] -1-methoxyoxaloyl-azetidin-2-one] 80 0 40 oco2pnb oco2pnb

SVxj | ^^ S'0TBDMS OTBDMS

cdoCH ^. COOCH ^ 0.7 g of 4 (R) - {1-tert-butyldimethylsilyloxymethylvinylthio) -3 (S) - [T (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-methoxycarbonyl-2-methyl- 1-Propenyl) -azetidin-2-one was dissolved in a mixture of 30 ml of dichloromethane and 10 ml of methanol. The solution was cooled to -78 ° C, ozone in oxygen was blown through the solution until a blue color appeared. After shaking with aqueous solution and drying over sodium sulfate and evaporation of the solvent, 0.6 g of the title compound was obtained.

Example 12: 4 (R) -tert-Butyldimethylsilyloxyacetylthio-3 (S) - [7 (R) -p-nitrobenzyloxycarbonyloxyethyl] azetidin-2-one

OCO-PNB

Λ “H fip0,” ®

-E- ^ OTEDHS

^ 1 · - * JlOi COOCH ^ 0 ίο 80040 0.6 g of 4 (R) -tert-butyldimethylsilyloxyacetylthio-3 (S) - [(R) -p-nitrobenzyloxycarbonyloxyethyl] -1-methoxyoxaloyl-azetidine- The 2-one was dissolved in 30 ml of methanol and a few grams of silica gel were added. After stirring for 1 h, the mixture was filtered and the solvent was evaporated from the filtrate. Chromatography of the residue on silica gel using cyclohexane-ethyl acetate (3: 2) as eluent gave 0.28 g of the title compound.

PMR (CDCl 3) δ (ppm): 0.15 (s, 6H, Si (CH)), 0.95 (s, 9H,

SiCl 2 CH 2), 1.45 (d, J = 6.5 Hz, 3H, CH 3 CH), 3.42 (dd, J = 3.0, 6.0 Hz, 1H, H-6, 4.25 (s, 2H, CH OSi), 5.26 (s, 2H, CH 2 Ph), 5.1-5.3 (m, 2H, CHCH 3, H-5), 6.70 (broad s, 1H, NH, 7.4-8 .4 (m, 4H, Ph, NO).

1R (CH 2 Cl 2): u (cm-1): 169S C = 0, 1750 -OCC-, 1785 β-lactan

Example 13: 4 (R) -tert-Butyldimethylsilyloxyacetylthio-3 (S) - [T (R) -p-nitrobenzyloxycarbonyl-oxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-hydroxymethyl) -azetidine-2- one

OCO ^ PNB OCC ^ PNB

0TBDMS

C02PNB

A mixture of 0.34 g of 4 (R) -tert-butyldimethylsilyloxyacetylthio-3 (S) -dis (R) -p-nitrobenzyloxycarbonyloxyethyl] azetidin-2-one and 0.34 g of p-nitro benzyl glyoxylate in 10 ml of benzene, was kept at reflux for 2 hours. After evaporation of the solvent, the residue was purified by silica gel column chromatography using cyclohexane-ethyl acetate (3: 2) as the eluent to give 0.27 g of the title compound.

PMR (CDCl 3; δ (ppm): 0.13 (s, 6H, Si (CH)), (s, 9H,

SiC (CH) -), 1.47 (d, J = 6.5 Hz, 3H, CH 2 CH), 3.52 (m, 1H, H-6), -2-3-3-27 ( s, 2H, CH 2 Cl 2), 4.0-4.6 (m, 2H, CHOH, CHOH), 5.25 (6.4H, two CH 2 Ph), 5.1-5.6 (m, 2H, CHCH 2, H-5), 7.3-8.3 (m, 8H, two Ph-NO 2).

Example 14; 4 (R) -tert-Butyldimethylsilyloxyacetylthio-3 (S) - [T (R) -p-nitrobenzyloxycarbonyl-oxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-chloromethyl) -azetidin-2-one oco2pnb oco2pnb

A solution of 0.27 g of 4 (R) -tert-butyldimethylsilyloxyacetylthio-3 (S) - [(R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-hydroxymethyl) -azetidin-2-one in 3 ml of anhydrous tetrahydrofuran, cooled to 0 ° C. 0.045 ml of pyridine and 0.03 ml of thionyl chloride were added. After 10 minutes, the mixture was filtered.

Evaporation of the solvent gave 0.3 g of the title compound, which was used as such in the next step.

2o 80040

Example 15: 4 (R) -tert-Butyldimethylsilyloxyacetylthio-3 (S) - [(R) -p-nitrobenzyloxycarbonyloxyethyl] -1 '(1-p-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl) azetidine- 2-one

OCO ^ NB OCOjPNB

j (t> —Cl ° J% = PPh-

CO PNB 0 CO PNB J

z <0.3 g of 4 (R) -tert-butyldimethylsilyloxyacetyl-thio-3 (S) - [} (R) -p-nitrobenzyloxycarbonyloxyethyl? ” 1- (1-p-Nitrobenzyloxycarbonyl-1-chloromethyl) -azetidin-2-one and 0.45 g of triphenylphosphine were dissolved in 5 ml of dichloromethane, and 2-3 g of silica gel was added to the solution. After evaporation of the solvent, the added silica gel was dried, left overnight at room temperature and washed with cyclohexane to remove excess triphenylphosphine. The product adsorbed on diatomaceous earth was chromatographed on silica gel using cyclohexane-ethyl acetate (3: 2) as eluent. There was thus obtained 0.26 g of the title compound.

PMR, CDCl 3) δ (ppm): 0.0c », 0.15 (two S, 6H, SiiiCH 2, 0.8S 0.93 (two s, 9H, SICiCH 2), 1.35 (d, J = 6.5 Hz, 3H, CH 2 CH 4), 4.1-4.2 (m, 2H, CH 2 OH), 4.6-5.0 (m, 1H, CHCH 3), 5.20 (broad s, 4H, two CH 2 -Ph-NO 2), 7.56 (broad s, 15H, P (Ph) 3) δ 7.6-8.4 (m, 8H, broad Ph-NO 2).

Example 16: 4 (R) -Hydroxyacetylthio-3 (S) -N- (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-i-triphenylphosphoranylidene)

II

2i B0040

methyl) -azetidin-2-one

COPNB

2

Solution of 0.26 4 (R) -tert-butyldimethylsilyloxyacetylthio-3 (S) - [T (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl) -azetidin-2-one and 0.07 ml of acetic acid in 2 ml of anhydrous tetrahydrofuran were treated with a solution of 0.18 g of tetrabutylammonium fluoride in 2 ml of tetrahydrofuran. After stirring at room temperature for 1 h, the mixture was diluted with ethyl acetate, washed with water, saturated sodium bicarbonate solution and again with water.

After drying and evaporation of the solvent, the residue was purified by silica gel column chromatography using cyclohexane-ethyl acetate (1: 3) as the eluent to give 0.13 g of the title compound.

PMR (CDCl 3) δ (ppm): 1.37 (d, J = 6.5Hz, 3H, CH 3 CH), 4.2 (n, 2H, CH 2 OH), 4.9 (m, 1H, CH 3 CH), δ, 25 (m, 5H, two CH 2 Ph, H-5, 7.55 (s, 15H, P (Ph) 3), 7.6-8.4 (m, 8H, two PhNQ 2).

Example 17; p-Nitrobenzyl (5R) -2-hydroxymethyl-6 (S) - {[(R) -p-nitrobenzyloxycarbonyloxyethyl] -2-penem-3-carboxylate 22 80040 oco2pnb 0CO2pnb -> s'Y ^ ch2oh 0 / = ^ 3 <r * δο, ΡΝΒ CO PNB 2 2

A solution of 0.13 g of 4 (R) -hydroxyacetylthio-3 (S) - [(R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl) -azetidine-2- on 10 ml of xylene, was refluxed for 1 h under a nitrogen atmosphere. Evaporation of the solvent and purification of the residue by preparative thin layer chromatography (silica gel) gave 50 mg of the title compound.

C 32 O + 66 ° (c = 1.3 CHCl 3) PMR; CDCl 3) δ (ppm): 1.51 (d, J = 6.5, 3H, CH 3 CH), 3.55 (broad s, 1H, OH), 3.97 (dd, J = 2.0, 8.0 Hz, 1H, H-6), 4.68 (s, 2H, CH 2 OH), 5.19 (dq, J = 6.5, 8.0Hz) , 1H, CHCH 3), 5.25-5.45 (m, 4H, two CH 2 Ph), 5.65 (d, J = 2 Hz, 1H, H-5), 7.4-8.5 ( tn, 8H, two PhNO 2).

Mass spectrum (F.P.) m / e 559 UV: λ (CH Cl): 269 nm (£ 17,000), 323 fc6800) max d 2 ΙΪ! (CH 2 Cl 2) (cm-1): 1795, 1755, 1710.

Example 18: p-Nitrobenzyl (5R) -2-tert-butyldimethylsilyloxymethyl-6 (S) (R) -p-nitroxyloxycarbonyloxyethyl] -2-penem-3-carboxylate 23 80040 CCO FNB „OMT3

f 2 0CO2PNB

^ - [| T OTBDHS ^ ^ OTBDMS

J N \ _ * -N-L

0 / Pi% w CO PNB

o 2

CO PNB

M

A solution of 0.15 g of 4 (R) -tert-butyldimethylsilyloxyacetylthio-3 (S) (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl) azetidine- 2-one in 15 ml of xylene was stirred for 1 hour under nitrogen at reflux. When the solvent was evaporated and the residue was purified by preparative thin layer chromatography on silica gel, 70 mg of the title compound were obtained.

Example 19: p-Nitrobenzyl (5R) -2-hydroxymethyl-6 (S) - (λ (R) -p-nitrobenzyloxycarbonyloxyethyl] -2-penem-3-carboxylate

OCOjPNB 0CO2PNB

(Γ "ko2PNB ko2PKB

70 mg of p-nitrobenzyl (5R) -2-tert-butyldimethylsilyloxymethyl-6 (S) - (Λ (R) -p-nitrobenzyloxycarbonyloxyethyl) -2-penem-3-carboxylate was dissolved in anhydrous tetrahydrofuran, and 0.025 ml was added. acetic acid and a solution of 68 mg of tetrabutylamino-2,80040 monium fluoride in 0.5 ml of tetrahydrofuran The mixture was stirred for 1 h at room temperature, diluted with ethyl acetate and washed with water, saturated sodium bicarbonate solution and again with water. thin layer chromatography on silica gel using cyclohexane-ethyl acetate (3: 7) as eluent gave 30 mg of the title compound, which was identical to that obtained in Example 17 (IR and NMR spectra).

Example 20: p-Nitrobenzyl (5R) -2- (N-trichloroacetylcarbamoyloxymethyl) -6 (S) -N- (R) -p-nitrobenzyloxycarbonyloxyethyl) -2-penem-3-carboxylate

OCOoPNB

Λ · 1 ——- P-1

2 O CO2PNB

To a solution of 50 mg of p-nitrobenzyl (5R) -2-hydroxymethyl-6 (S) - [(R) -p-nitrobenzyloxycarbonyloxyethyl) -2-penem-3-carboxylate in 1 ml of purified acetone 0 At 0 ° C, a solution of 0.06 ml of trichloroacetyl isocyanate in 1 ml of purified acetone was added dropwise. After 20 minutes, the solvent was evaporated to give 100 mg of the title compound.

OYR (CDCl 3) δ (ppm): 1.50 (d, J = 6.0Hz, 3H, CH 2 CH), 4.00 (dd, J = 2.0, 8.0Hz, 1H, H-6) , 5.1-5.9 (m, 8H, H-5, CHO), two CH 2 Ph, CH 2 OCO), 7.5-8.4 (m, 8H, two PhNO 2), 8.90 (broad s, 1H, NH), li 25 80040

Example 21: p-Nitrobenzyl (5R) -2-carbamoyloxymethyl-6 (S) - {λ (R) -p-nitrobenzyloxycarbonyloxyethyl] -2-penem-3-carboxylate oco2pnb oco2pnb

^ KONHCOCClj l- / s \ / \ OCOHH

J N \ J'-H-

0 CO PNB O CO PNB

2 l of 100 mg of crude p-nitrobenzyl (5R) -2- (N-trichloroacetylcarbamoyloxymethyl) -6 (S) - [(R) -p-nitrobenzyloxycarbonyloxyethyl] -2-penem-3-carboxylate was dissolved in 4 ml of methanol. Silica gel (40-63 μm) was added and the mixture was stirred for 3 hours at room temperature and filtered. Washed with acetone. After evaporation of the solvent from the filtrate, the residue was purified by preparative thin layer chromatography on silica gel using cyclohexane-ethyl acetate (3: 7) as the eluent to give 33 mg of the title compound.

= + 50 ° (c = 2.4, acetone) PMR (CDCl 3) δ (ppm): 1.48 (d, J = 6.5Hz, 3H, CH 2 CH), 3.95 (dd, J = 2 0.08 Hz, 1H, H-6), 4.85 (broad s, 2H, NH4)> 5.1-5.5 (m, 7H, CHCH2, two CH2Cl2, CH2CO), 64 (d, J = 2.0 Hz, 1H, H-5).

7.4-8.5 (m, RH, two PhNO 2) IR (KBr) 1 J (cm -1): 1795, 1750, 1710.

Example 22: 4 (R) - (N-Trichloroacetylcarbamoyloxyacetylthio) -3 (S) - [(R) -p-nitrobenzyloxycarbonyloxyethyl] -1 '(1-p-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl) azetidine -2-one 26 80040

0ρ ° 2ρΝβ OCOoPKB

irf5 Τ ^ Η / ^ P ^ Sv | X ^ tcoKHcccci3 / ~~ r? h3 <y% = pph,

CO PNB CO PNB J

1 2 120 mg of 4 (R) -hydroxyacetylthio-3 (S) - [1 (R) -p-nitrobenzyloxycarbonyloxyethyl] -1-n-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl) azetidin-2-one were dissolved in 2 ml of: purified acetone and cooled to 0. A solution of 0.1 ml of trichloroacetyl isocyanate in 2 ml of purified acetone was added dropwise, and the resulting mixture was stirred for 1/2 hour.

Evaporation of the solvent gave 180 g of crude title compound.

Example 23: 4 (R) -Carbamoyloxyacetylthio-3 (S) - [T (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidenemethyl) azetidin-2-one ffV® cco2pkb XONHCOCCX3 -yS'Y ^ OCONH2 cf / = PPh. 0 / *> 0 ^> = .pph CO PNB / 3

2 CO 2 PNB

M

A mixture of 180 mg of crude 4 (R) - (N-trichloroacetylcarbamoyloxyacetylthio) -3 (S) - [(R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-triphenylphosphoranylidene methyl) ) -azetidin-2-one in 8 ml of methanol and

II

2? 80 0 40 lickels (40-63 μπι) were stirred for 4 hours at room temperature. The mixture was filtered, washed with acetone and the filtrate evaporated to dryness. The residue was purified by preparative thin layer chromatography using cyclohexane-ethyl acetate (1: 4) as eluent to give 70 mg of the title compound.

Example 24; p-Nitrobenzyl (5R) -2-carbamoyloxymethyl-6 (S) - [7 (R) -p-nitrobenzyloxycarbonyloxyethyl] -2-penem-3-carboxylate oco2pkb oco2phb

-f S'Y ^ 0C0HH2 f ^ V ^ OCCNH

J — l e ——— ► I _ [_ Il

^> = PPh3 ζ CO ^ NB

CO PKB 2

A solution of 70 mg of 4 (R) -carbamoyloxyacetylthio-3 (S) (R) -p-nitrobenzyloxycarbonyloxyethyl] -1- (1-p-nitrobenzyloxycarbonyl-1-triphenyl-phosphoranylidenemethyl) -azetidine-2- on 8 ml of xylene, was stirred for 1 hour under a nitrogen atmosphere at reflux temperature. Evaporation of the solvent and purification of the residue by preparative thin layer chromatography on silica gel gave 30 mg of the title compound identical to the product obtained in Example 21 (IR and NMR spectra).

Example 25: Sodium (5R) -2-carbamoyloxymethyl-6 (S) - [Ϊ (R) -hydroxyethyl] -2-penem-3-carboxylate 28 80 0 40 0CO 2 PNB oh - | OCOHH, Q rCO PHB O CC ^ Na® 30 mg p-nitrobenzyl- (5R) -2-carbamoyloxymethyl-6 (S) - [(R) -p-nitrobenzyloxycarbonyloxyethyl] -2-penem The 3-carboxylate was dissolved in 3 ml of ethyl acetate. 2 ml of water, 4.2 mg of sodium hydrogen carbonate and 45 mg of 5% Pd / C were added and the mixture was hydrogenated at room temperature for 2 h. Filtered through diatomaceous earth and the aqueous phase washed with a small amount of cold ethyl acetate, filtered through Waters Sep-Pak Cj g and freeze-dried. The residue was purified by reflux chromatography on Waters Sep-Pak g tubes using water as eluent. 8 mg of the title compound were obtained.

UV: λ (H_ =): 259 nm (£ 3500), 308 (5400) max λ PMR (DO) δ (ppm): 1.31 (d, J »6.5Hz, 3H, CH 2 CH), δ .91 (dd, J = 1.5, 6.0Hz, 1H, H-6), 4.25 (m, 1H, CHOH), 5.02, 5.36 (two d, 2H, CH 2 OCO), δ .66 (d, J = 1.5 Hz, 1H, H-5).

{* 3 * ° = + 143 ° (c = .97, H 2 O).

II

Claims (2)

29 80 0 40 CLAIM Analogue method of formula OH ÄLvx I | II) For the preparation of a compound having antibacterial activity according to CONH2 (1} <J ^ COOH or a pharmaceutically acceptable salt thereof, characterized in that the compound fr 1 COOK of general formula II wherein R is p-nitrobenzyloxycarbonyl, dimethyl-tert-butylsilyl, diphenyl -tert-butylsilyl, 2,2,2-trichloroethoxycarbonyl, trimethylsilyl, benzyl, p-bromophenacyl, triphenylmethyl or pyranyl and R is lower alkyl, 2,2,2-trichloroethyl, acetonyl, allyl, benzyl , p-nitrobenzyl, p-methoxybenzyl, phenyl, o-nitrophenyl, benzhydryl or 1-phenoxyethyl, the hydroxy group is protected with a protecting group PG which is e.g. p-nitrobenzyloxycarbonyl, dimethyl-1-tert-butylsilyl, diphenyl -silyl, 2,2,2, -trichloroethoxycarbonyl, trimethylsilyl, benzyl, p-bromophenyl, triphenylmethyl or pyranyl to give a compound of formula III 80040 ...... COCR wherein R, R and PG are the same as above, pel phosphorus tribromide11a to give the compound of formula IV QR ..... wherein R, R1 and PG have the same meaning as above, is ozonolysed in a solvent at -80 to -50 ° C to give the compound of formula IVa 0R1 τςτ - ...... COOR wherein R, R and PG are as defined above, methanolized in the presence of silica gel to give a compound of formula V OR wherein R and PG are as defined above, condensed at a temperature of 70-100 ° C With a compound of formula CHOCOOR, where R is as defined above, that the compound of formula X obtained is HH O (X) / -γ COOR wherein R1 and PG are as defined above, chlorinated with thionyl chloride di Compound of formula XI · <χι> / γ COOR wherein R, R and PG have the same meaning as above, is reacted with triphenylphosphine at a temperature of 30-60 ° C, and that the compound of formula XII obtained Yo- ...... ° COOR wherein R, R 1 and PG have the same meaning as above, m is extracted into a compound of formula I by carrying out the following reaction steps (a) to (c) in the order shown, or so that step (a) is carried out after step (b) but before step (c), or so that step (a) is carried out in steps after (b) and (c), or when the group PG is hydrogen, step (b) is omitted and step (a) is performed before or after step (c): (a) ring formation is performed by heating in a nitrogen atmosphere in an inert solvent at a temperature in the range of 80 -150 ° C, (b) deprotecting the substituent of the following formula with 32 80 0 40 ° trifluoroacetic acid in ethyl acetate or tetrabutylammonium fluoride in tetrahydrofuran, optionally in the presence of acetic acid. (c) attaching a carbamoyl group to the resulting substituent of the following formula ΥΥ by reaction with trichloroacetyl isocyanate in acetone and treatment with silica gel in methanol, and finally deprotection to give a compound of formula I which is optionally salified. Il 33 80 040 Analogous pharmaceutical formulation with an antibacterial formulation of the formulation (I) or a pharmaceutical formulation site, OH ΛΙ I j || (O) g of COOH can be converted to compounds of formula II and 'KT' * "" COOR from R1 to p-nitrobenzyloxycarbonyl, dimethyl-tert-butyl-silyl, diphenyl-tert. -butylsilyl, 2,2,2-trichloroethoxycarbonyl, trimethylsilyl, benzyl, p-bromophenyl, triphenylmethyl or pyranyl, oohr is alkyl, 2,2,2-trichloroethyl, acetonyl, allyl, benzyl, p-nitrobenzyl, p-methoxybenzyl, phenyl, o-nitrophenyl, benzhydryl or 1-phenoxyethyl, hydroxyl group with a group PG, as is t.ex. p-nitrobenzyloxycarbonyl, dimethyl-tert-butylsilyl, diphenyl-tert-butylsilyl, 2,2,2-trichloroethoxycarbonyl, trimethylsilyl, benzyl, p-bromophenyl, triphenylmethyl or pyranyl, to give 34% of this form ! T ^ r- - COCR of R, Ri and PG of decamma ovan, reducers with phosphorus tribromides, to give the same particulars of formula IV -N — rys> Y''sN * 6 / V COOR of R, Ri and PG above the ozone, the ozonolysis is carried out at a temperature of -80 to -50 [deg.] C., at the same time as the formulation IVa ORj 7 = ζΓ ~ ~ COOR for R, R ^ och and PG on silica gel, preferably in the form of V orj 35 8 0 0 4 0 from R, Rj and PG at a temperature of more than 70-100 ° C The temperature in the case of CHOCOOR, at a temperature of 70-100 ° C ovan, to give an enriched form X - Rih '^ tY * sV ^ pg (X, COOR of R, R 1 and PG of decanted soran ovan, chlorinated Raed thionyl chloride, of which an exemplified form of exogenous form XI -. 0 T COOR där R, R 1 and PG are further selected from the group consisting of triphenylphosphine at a temperature of 30-60 ° C, and preferably at room temperature. aeln XII i ^> Y ^ n's ° pg I o (xii) /-K.™* 0 COOR r R, Ri och PG aver decamna sora ovan, förändras account en förening enligt formuleln I genora att utföra feljandeeactionskeden (a) - (c) i den ordning sora de presenteras eller sä, where skedet (a) utförs efter skede (b) men for skedet (c), eller sä, att skedet (a) utförs efter skedena (b) och 36 8 0040 (c), eller dä Gruppen PG är väte, skedet (b) sloppy wine and skedet <a (c): (a) the cyclization of the genome is carried out in an ambient atmosphere at an inert temperature of 80 to 150 ° C; ammonium fluoride in tetrahydrofuran, optionally under the influence of acetic acid, (c) the carbamoyl group is substituted with a substituent or a formulation of the reaction with trichloroacetyl socyanate in the form of acetone and with the addition of a mixture of methanol and hydrogen, e.g. I erhälles, son dä sä önskas föränd-ras till sitt salt.