FR2491071A1 - ESTERS OF 2-B-CHLOROMETHYL-2-METHYLPENAME-3-A-CARBOXYLIC ACID SULFONE AND A PENICILLIN, PROCESSES FOR THEIR PREPARATION AND PHARMACEUTICAL COMPOSITIONS CONTAINING THE SAME - Google Patents

Abstract

THE PRESENT INVENTION CONCERNS THE PHARMACEUTICAL INDUSTRY. IT HAS AS A SUBJECT COMPOUNDS WITH ANTIBIOTIC ACTIVITY RESPONDING TO THE FORMULA: (CF DRAWING IN BOPI) THE INVENTION ALSO CONCERNS PROCESSES FOR PREPARING THESE BODIES.

Description

The present invention relates to sulfone

  of 2β-chloromethyl-2α-methylpena-3α-carboxylic acid

  a pharmaceutically acceptable salt or a readily hydrolysed ester thereof which is useful as a 3-lactamase inhibitor. The presumed association between the resistance of 3-lactam antibiotics to certain bacteria and the ability of these bacteria to produce 3-lactamases has led to a

  intensive search for 3-lactamase inhibitors.

  Clavulanic acid is an example of such a compound currently studied in detail. Another β-lactamase inhibitor responds in acid form to the formula:

H 2 CH

  -H2H H3 COOH and is the subject of the European patent application 2927

published on July 11, 1979.

The compound of formula: 277 H2c

H N 2 C

O * COOH

  is described in U.S. Patent Nos. 4,036,847, 4,009. 159, 5,995,646, 3,989,685 and

5954752.

  The patent of the United States of America

  No. 4,155,912 discloses 2-penem-3-carboxylic acid derivatives.

  Boxylic acid of the formula: ## STR5 ## as well as esters and salts, about which reference can also be made to Farmdoc Abstracts

82090A, 10336B and 4-43537B.

  The compound (No. CP-45899) of

  the: Qo. -O + S -t H3 .CH3 COOH, is an irreversible 3-lactamase inhibitor having excellent stability in solution. He has one

  weak antibacterial activity and potentiates the

  In Vitro and In Vivo Experiences of Ampicillin for Strains Producing 3-Lactamase [A. R. English et al., Antimicrobial Agents and Chemotherapy, 14, 414-419 (1978), Aswapokee et al., J. Antibiotics 31 (12), 1238-1244 (Dec. 1978) and Derwent's Farmdoc Abstracts.

89627 and 73866B].

  It was reported by B. Baltzer et al., "Mutual

  Pro-Drugs of P-Lactam Antibiotics and P-Lactamase Inhibit

  bitors, J. Antibiotics, 33 (10), 1183-1192 (1980) that the

  principle of combining an antibiotic with a P-lac-

  tame with a β-lactamase inhibitor in a single molecule functioning as a prodrug for both active constituents is illustrated by the esters

  combined 3 and 4 in which ampicillin and

  liname, respectively, are combined with sulfone

  of penicillanic acid which is a P-lactam

  mase. It is shown that in humans these esters are

  absorbed in the gastrointestinal tract

  intestinal tract and hydrolysed after absorption with

  simultaneous application of the active ingredients. As a result, high blood and tissue levels are achieved in

  antibiotic and as a 0-lactamase inhibitor

  feel a balanced relationship. The advantages offered by the "precursors of mutual medicines" on the

  simple combinations are envisaged.

  The esters 3 and 4 mentioned are those

  with the formulas: o0 CO CH -0 Co CH CO-O

20 .....

  published the ring of the general: o30 It is indicated in the document GB 2044255

  October 1980 that the invention described

  hitherto unknown compounds of formula

R1-CH-CO-NH Y

R2 s s o H

H C-O-CH-A

  R 1 represents a phenyl, 4-hydroxyphenyl, 1,4cyclohexadienyl or 3-thienyl radical; R2 represents a primary amino or carboxyl radical; R3 represents a hydrogen atom or a lower alkyl, aryl or aralkyl radical and A represents a radical of a β-lactam-containing β-lactamase inhibitor, in addition to a carboxyl radical, A being united via the carboxyl radical. These new compounds are useful for

  treatment of bacterial infections and are, in

  particular, highly active against the bacteria produced

  O-lactamase. We can refer to this topic

  also at Farmdoc Abstracts 60773C and 60776C.

  The subject of the invention is therefore the acid of

mule:

O O

S CH2C1

3

  or a pharmaceutically acceptable salt of acid or

  an easily hydrolysed ester of the acid.

  The pharmaceutically acceptable salts men-

  above are non-toxic metal salts, such as sodium, potassium, calcium and magnesium salts, ammonium salt and salts.

  substituted ammonium compounds, such as non-toxic amine

  such as trialkylamines (for example

  ethylamine), procasin, dibenzylamine, N-ben-

  3-phenethylamine, 1-ephenamine, N, N'-dibenzene

  zylethylenediamine, dehydroabietylamine, N, N'-

  bis (dehydroabiethyl) ethylenediamine, the N-alkyl

  piperidines (eg N-ethylpiperidine) and other amines which have been used to form pharmaceutically acceptable salts of penicillins and cephalosporins. Especially preferred salts are the alkali metal salts, i.e.

  sodium and potassium, in addition to ammonium salt.

  For purposes of the invention, by "physiological esters

  logically hydrolysed "means the pharmaceutically acceptable esters of which we know

  that they hydrolyze into free acids in vivo. Examples

  Suitable esters of physiologically hydrolysed esters are esters comprising a phenacyl radical,

  toxymethyl, pivaloyloxymethyl, -acetoxyethyl, a-

  acetoxybenzyl, α-pivaloyloxyethyl, phthalidyl (3-phthalic

  dyl), indanyl (5-indanyl), methoxymethyl, benzoyloxy

  methyl, α-ethylbutyryloxymethyl, propionyloxymethyl,

  10 valeryloxymethyl, isobutyryloxymethyl, 6 - [(R) -2-amino-

  2-phenylacetamido] -3,3-dimethyl-7-oxo-4-thia-l-azabicy-

  cloE3.2.0] heptane-2-carbonyloxymethyl and 6 - [(R) -2-amine

  no-2-p-hydroxyphenylacetamido] -3,3-dimethyl-7-oxo-4-

  thia-azabicyclo [3.2.0] heptane-2-carbonyloxymethyl.

  The preferred esters are the esters comprising a radical

  acetoxymethyl, pivaloyloxymethyl, methoxymethyl,

  phthalidyl, 5-indanyl, 6 - [(R) -2-amino-2-phenylacetamide

  do] -3,3-dimethyl-7-oxo-4-thia-l-azabicyclo [3.2.0] hepta-

  ne-2-carbonyloxymethyl and 6 - [(R)] - 2-amino-2-p-hydroxy-

  phenylacetamido] -3,3-dimethyl-7-oxo-4-thia-l-azabicy-

  clo [3.2.0] heptane-2-carbonyloxymethyl. The invention also relates to the process for producing the desired acid of formula: If CH2C1

00, 2

  CH 3 OOH or a pharmaceutically acceptable salt thereof, in which, at successive stages, a) the catalytic hydrogenation, for example using a catalytic precious metal such as palladium, an ester of formula: A o R1 represents a kill;

benzyl or substituted benzyl radical

  b) the hydrogenated product is subjected to oxidation to form the desired acid or its salt, then, if desired, c) the acid or salt is esterified to produce

  an easily hydrolysed ester of the acid.

  The subject of the invention is furthermore the process for producing the desired acid of formula: ## STR1 ## or a pharmaceutically acceptable salt of this acid, according to which, at successive stages, a) is oxidized, for example by means of KMnO 4, of H202 or a similar peroxide or a peracid, an ester of formula: -Br kCH2C1

O C 'C

w1C1

C- OCH2CCJ3

  O to form an ester sulfoxide of formula: ## STR3 ## / CH2

"C H3

O C-OCH2CCL,

  o then b) this ester sulfoxide is reacted with a

  metal in an acid, such as zinc in acetic acid

  than cold to produce the desired acid or salt, then, if desired, c) the acid or its salt is esterified to produce

  an easily hydrolysed ester of the acid.

  Many oxidants known for the oxidation of sulfides to sulfones are suitable. Nevertheless,

  Particularly suitable reagents are permanga-

  alkali metal compounds, such as the permanganate of

  potassium, and organic peracids, such as 3-

  chloroperbenzoic. Particularly useful protective groups 1 are the benzyl radical and the benzyl radicals

  substituted and especially the 4-nitrobenzyl radical.

  Benzyl and substituted benzyl radicals can be

  conveniently removed by catalytic hydrogenation.

  In this case, a solution of the compound of formula A,

1 -

  R represents a benzyl or substituted benzyl radical,

  in an inert solvent is stirred in an atmosphere of

  or with hydrogen mixed with a di-

  inert luent such as nitrogen or argon, in the presence

  catalytic amount of a hydrogen catalyst

  nation. Solvents suitable for such a hydrogen

  are lower alkanols, such as methanol, ethers, such as tetrahydrofuran and dioxane; low molecular weight esters, such as ethyl acetate and butyl acetate, water and mixtures of these various solvents. However, it is usual to choose conditions in which the compound of

  starting is soluble. Hydrogenation is carried out

  at a temperature of about 0.degree. to 600.degree. C. under a pressure of about 101 to 10100 kPa. The catalysts used for this hydrogenation are of the type known for such a transformation, typical examples being noble metals, such as nickel, palladium,

  platinum and rhodium. The catalyst is usually

  in an amount of about 0.01 to 2.5% by weight and preferably about 0.1 to 1.0% by weight, based on the compound of formula A. It is often advantageous to deposit the catalyst on an inert support and a particularly convenient catalyst is palladium deposited on an inert support such as carbon. Of

  more, it is usual to dab the reaction mixture

  to work at a pH of about 4 to 9 and,

  preferably from 6 to 8. Borate and phosphorus buffers

  phate are in common use. The reaction is normal

in about 1 hour.

  The invention further relates to esters of formula: CH.

R - CH - CO - NH M

  Wherein R represents: R2 * i. Ri

  R represents a hydrogen atom or a hydrogen radical;

  and R represents a hydrogen or chlorine atom or a hydroxyl, methyl or methoxy radical, and preferably the ester of formula: ## STR2 ##

HO CH-CO-NH

NH2 2 o OS '\ C

C 0 O

  An ester of formula: NH 2

  as well as the process for producing such an ester,

  wherein an acid is reacted with a solution of a compound of the formula: ## STR1 ## wherein R represents

ZYKYO or R4O.

  R represents a hydrogen atom or a hydrogen radical;

  and R5 represents a hydrogen or chlorine atom or a hydroxyl, methyl or methoxy radical; R1 represents an alkyl, aralkyl or aryl radical; R 2 represents a hydrogen atom or an alkyl, aralkyl or aryl radical and R 3 represents an alkyl, aralkyl, aryl, alkoxy, aralkoxy, aryloxy or NR 4 R 5 radical. R 4 and R 5 each represent a hydrogen atom or

  an alkyl, aralkyl or aryl radical or taken

  with the nitrogen atom, represent a piperidino or morpholino radical, which reaction with the acid is preferably carried out in an organic solvent such as acetone or chloroform or in an aqueous or partially aqueous solution and of

  preferably at a pH of 1 to 5 at room temperature.

  It is furthermore preferred that in the formula III of the radical protecting the amino function, R 1 represents A t R 2 a methyl radical, R 2 represents a hydrogen atom and R 3 represents a methoxy, ethoxy or methyl radical,

  which requires the use of methyl acetoacetate

  1α, ethyl acetoacetate or acetylacetone. For the removal of the protective radical of the amino function, it is preferable to use a strong mineral acid, such as hydrochloric acid, or else

formic acid.

  The subject of the invention is also, as a novel intermediate, an ester of formula: Br S CH2Cl

3

  -OR o R represents a substituted benzyl or benzyl radical and preferably p-nitrobenzyl, as well as the process for preparing this ester, according to which a compound of formula: Br OC-OR or R represents, preferably at reflux, a benzyl or substituted benzyl radical

  and preferably p-nitrobenzyl, in an organic solvent.

  inert anhydride, preferably dioxane, in

  the presence of a significant quantity and preferably

  molar a weak tertiary amine, preferably quinoline, and an acid chloride, preferably benzoyl chloride, until substantial completion of the reaction. The invention furthermore relates, as a novel intermediate, to an ester of formula: ## STR1 ## where R represents a benzyl or substituted benzyl radical and preferably p-nitrobenzyl, as well as the process for preparing this ester, according to which an solution, in an inert solvent, preferably methylene chloride, of a compound of the formula: ## STR1 ## wherein R represents a substituted benzyl or benzyl radical;

  and preferably p-nitrobenzyl, at about the same temperature as

  ambient, by means of a peracid, preferably

M-chloroperoxybenzoic acid.

  The subject of the invention is also, as a novel intermediate, the ester of formula: ## STR1 ## as well as the process for preparing it, according to which

  a solution is oxidized in an inert solvent, preferably

  methylene chloride, a compound of formula

the: Br C H2Cl

0 CHC3

NOT, ,

  C-OC H2CCl3 o

  at about room temperature with an oxy-

  such as potassium permanganate, hydrogen peroxide or a similar peroxide or preferably a

  peracid, advantageously m-chloroperoxybenzol-

  than. "Skellysolve B" is a fraction of ether

  oil boiling from 60 to 68 C and consisting essen-

  typically n-hexane, sold by Skelly Oil Co.

  Examples below illustrate the preparation of

  representative compounds which are the subject of

  the invention without limiting the latter.

EXAMPLE.l o

  Preparation of 2-chloromethyl-2 "-methyl sulfone

  potassium pena-3a-carboxylate (BL-P2013)

O O

  3 ... Br "v XNO 2 o N" CO ... H N --- _ _ Br Br NO2 Mnn04

(BL-P2013)

  6-bromopenicillanic acid S-sulfoxide (1)

  1. Dissolve 50 g (57.5 mmol) of the N, N'-

  6a-Bromopenicillanic acid dibenzylethylenediamine [G. Cignarella et al., J. Org. Chem. 27, 2668 (1962) and E. Evrard, Nature 201, 1124 (1964)] in 330 ml of

  methylene chloride. Stirred and cooled to 0 C.

  2. 13 ml (156 mmol) of concentrated hydrochloric acid is added to the solution of

  m6thylène. Precipitation of dibenzhydrochloride

  Zylethylenediamine is done in 1 minute. We shake the

  suspension at 0-5 C for 10 minutes.

  3. The precipitated amine hydrochloride is separated off

  by filtration on a pre-filtered filter of earth of

H2

4 1

  tomatoes (brand "Dicalite"). The filter cake is washed with 150 ml of methylene chloride. he

  Filtration must be completed as quickly as possible

  corn. The acidic solution in methylene chloride should not be stored for a long time. Filtration difficulties may arise from the fineness of the precipitate. A filter aid added to the

suspension may be useful.

  4. The filtrates of methyl chloride are mixed

  and washed with 60 ml of cold water. It is stirred for 5 minutes and the aqueous phase is discarded. The

  pH of the wash water is 2.0 to 2.3.

  5. Concentrate the 6-bromopedic acid solution

  Nicillanic acid in methylene chloride under

  reduced to a volume of 65 to 80 ml. We are

  cool the solution to 5 C with stirring.

  6. Under strong agitation, add cautiously

  13 ml (86.9 mmol) of 40% peracetic acid in half

  utes. The reaction is exothermic. By cooling

  in the ice bath, the temperature is maintained

  15 and 18 C. The sulfoxide begins to crystallize after addition of 10 ml of peracetic acid. We shake

  and the suspension is cooled to 0-5 ° C. for 2 hours.

  7. The suspension is filtered and the snow-white cake is washed in the following order: 10 ml of water at 5 ° C., ml of methylene chloride at 0 ° C. and finally 15 ml of heptane. 8. Dry the filter cake at 45 C in the air oven to constant weight, a period of 6 to 10 hours to be sufficient. A longer heater may cause a pink reflection. The weight of

  Compound 1 is 16.26 g is a yield of 73.24%.

  9. The state of the reaction mixture can be observed

  and the final product by layer chromatography

  in the toluene / 4 acetone / 1 acid systems.

  of acetic acid or 8 acetone / 8 methanol / 3 toluene / 1

  acetic acid. The final product is analyzed by

  nuclear magnetic resonance troscopy and

infrared troscopy.

  P-Nitrobenzyl 6α-bromopenicillinate S-sulfoxide (2) 7.5 g (41 mmol) of potassium 2-ethylhexanoate was added to a solution of 12 g (40 mmol) of 6-sulfur S-sulfoxide. -bromopenicillanic in ml of acetone. The salt is collected by filtration, washed with cold acetone and air-dried.

  to obtain a total quantity of 10 g. We say-

  the crystalline potassium salt in 75 ml of

  thylacetamide and 7.8 g (0.04 mol) of p-nitrobenzyl bromide were added to the solution. The solution is stirred at 23 ° C. for 24 hours. The mixture is diluted with 500 ml of water and extracted with ethyl acetate. The ethyl acetate extract is washed 4 times with water and dried over anhydrous magnesium sulfate. The solvent is evaporated at 350 ° C. (2000 Pa) in order to obtain a

  oil that crystallizes. The crystals are dispersed

  light compounds of the compound 2 in ether, then

  collected by filtration to obtain a quantity

  9 g (70 / o) RF. 124-125oC with decomposition.

  Analysis for C1515BrN2O6S Calculated: C, 41.98; H, 3.05; N, 6.52% Found: C, 42.00; H, 3.48; N, 6.98% Infrared spectrum (KBr): 1800 (F), 1740 (F), 1610 (b),

  1520 (F), 1450 (m), 1350 (F), 1060 (m), 740 (m) cm-1.

  Nuclear Magnetic Resonance Spectrum H (60 MHz, DMSO): δ 1.22 (s, 3H), 1.6 (s, 3H), 4.67 (s, 1H), 5.2 (d, JVI-5) Hz, 1H), 5.45 (s, 2H), 5.68 (d, J, 15 Hz, 1H),

7.5-8.5 (m, 4H).

  P-Nitrobenzyl chloromethyl-2-methyl-6-bromopenam-3-carboxylate (X) A solution of 5 g (12 mmol) of 6-bromopenicillanate S-sulfoxide is heated under reflux in a nitrogen atmosphere for 4 hours. of p-nitrobenzyl (2) in 120 ml of anhydrous dioxane in the presence of 1.5 g (12 mmol of quinoline and 1.6 g (12 mmol) of benzoyl chloride, dilute the solution with 600 ml of water and the ethyl acetate extract.

  the ethyl acetate extract is washed with a solution of

  5% sodium bicarbonate solution, 5% phosphoric acid solution and finally water. We dry

  organic layer over magnesium sulphate and

  evaporate at 35 ° C. (2000 Pa) to an oil. Crystal oil

  read and collect the crystals which are washed with ether and finally with cold toluene, to obtain 3.5 g

  (65%) of the compound 3 melting at 130-135 C with decomposition

  tion. Analysis for C₁HH₁ClClBrN₂OS Calculated: C, 40.06; H, 3.14; N, 6.23% Found: C, 40.19; H, 3.12; N, 6.75% Infrared spectrum (KBr): 1792 (F), 1740 (F), 1610 (f), 1520 (F), 1353 (F), 1280 (m), 1025 (f), 990 (f). ), 750 (f) cm-1 Nuclear Magnetic Resonance Spectrum (60 MHz, DMSO): S 1.45 (s, 3H), 3.5-4.3 (m, 2H), 5.05 (s, 1H), 5.42 (s, 2H),

  5.5 (d, 1.5 Hz, 1H), 5.62 (d, J "ul, 5 Hz, 1H), 7.5-

8.5 (m, 4H).

  2-Chloromethyl-2x-methylpenam-6α-carboxy-sulfoxide

  A solution of 1 g (2.2 mmol) of 3-chloromethyl-2-methyl-6-bromopenam-3α-c arboxylate is stirred.

  of p-nitrobenzyl (3) in 50 ml of methyl chloride

* with 473 mg (2.2 mmol) of m-chloroperoxy acid

  benzoic. The solution is stirred for 3 hours at 23 ° C. The methylene chloride is evaporated down to

  20 ml at 2000 Pa at 335C and dilute the concentrated solution.

  with 50 ml of heptane ("Skellysolve B"). Wave-

  the solvent and disperse the residue in

  ther to rapidly crystallize the compound (4) melting at 136137 C with decomposition and obtained in

  amount of 250 mg (24% yield).

  Analysis for C₁HH₁BrBrCl1N₂OSS Calculated: C, 38.68; H, 3.02; N, 6.02% Found: C, 39.14, -H, 3.13; N, 5.96% Infrared spectrum (KBr): 1800 (F), 1760 (F), 1520 (F), 1350 (F), 1200 (F), 1050 (m), 830 (f), 740 (f). ) cm-1. Nuclear Magnetic Resonance Spectrum H (60 MHz, DMSO): g 1.32 (s, 3H), 3.8-4.5 (m, 2H), 4.97 (s, 1H), 5.25 (d , J-v1, 5Hz, 1H), 5.45 (s, 2H), 5.6 (d, Jrvl, 5Hz, 1H),

7.8-8.5 (m, 4H).

  23-Chloromethyl-2α-methylpenam-3 "-carboxy-

  Potassium Latex (5) (BL-P2013) A suspension of 4 g of 30% palladium on diatomaceous earth ("ICelite" 1) and 2.8 g of sodium bicarbonate in 150 ml of water was added. to one

  solution of 7 g (15 mmol) of 20-chloro sulfoxide

  methyl-2 "-methyl-6" -bromopenam-3a-carboxylate from p-n

  trobenzyl (4) in 150 ml of ethyl acetate. We have

  mixed for 3 hours at 345 kPa.

  The catalyst is separated by filtration and collected

  the aqueous layer, to which 1.5 g of

  potassium ganate in 50 ml of water. We stir the

  1 hour and add 250 mg of

  fite of sodium. We filter the mixture and adjust the

  filtrate at pH 2 with concentrated hydrochloric acid.

  The solution is lyophilized to obtain an amorphous powder.

  white phe. The solid is extracted with ethyl acetate

  on, the extract is evaporated to 20 ml and diluted

  residue with 100-ml heptane ("Skellysolve B"). We are

  picks 20-chloromethyl-2acx-methyl acid sulfone

  white hygroscopic solid-3a-carboxylic acid.

  The acid is dissolved in acetone and solid potassium 2-ethylhexanoate is added thereto. A precipitate of a white crystalline salt is obtained which is collected by filtration to obtain 170 mg of the melting compound

  above 140 C with decomposition.

  Analysis for C8H. ClKN05S.2H2O Calc'd: C, 28.27; Hi, 3.24; N, 4.12% Found: C, 28.27; H, 3.69; N, 3.84% Infrared spectrum (KBr): 1790 (F), 1770 (m), 1620 (F), 1460 (m), 1370 (F), 1310 (F), 1200 (F), 1140 (F) ), 955 (m),

740 (m) cm -1.

  Nuclear Magnetic Resonance Spectrum H (10OMHz, D20): δ 1.68 (s, 3H), 3, 2-3.9 (m, ν v Hz, J ', 4 4 Hz, Jtv 6 Hz, 2H), 4.0-4.4 (m, 2H), 4.3 (s, 1H), 5.02 (dd, J v4 Hz),

JT 2 Hz, l 1H).

EXAMPLE 2

  2B-Chloromethyl-2-methylpena-3a-carboxy-2-sulphate

pivaloyloxymethyl late

  23-chloromeric acid sulfone is mixed with

  thyl-2α-methylpena-3α-carboxylic acid in dimethyl

  sulfoxide with 1 equivalent of triethylamine and stirred for dissolution. Then add

  1 equivalent of bromomethyl pivalate in dimethyl

  formamide. The resulting mixture is stirred at room temperature. The mixture is then clarified by filtration and the filtrate is poured into water.

  Ice. The solid solid is collected by filtration.

  washed with water and dried to obtain

the desired ester.

  The acetoxymeric radical esters are prepared

  thyl, methoxymethyl, acetonyl and phenacyl of the same

  by replacing, in the process above, the pivot

  bromomethyl late with an equimolar amount of

  chloromethyl cetyl ether, chloromethyl methyl ether

  chloroacetone and phenacyl bromide, respec-

tively.

EXAMPLE 3

  21-Chloromethyl-2α-methylpenam-3 "-carboxy-

  pivaloyloxymethyl late BL-P2024 O 0 eNs>, CH3 S C30 o - / JIjaCH2C1 + NaI + H20 + ClCH2-O-C-C (CH3) 3

10. O (306) Co2 -

(BL-P2013)

0 0 CH acetone

I _CH2C1

O C02CH2-0-C-C (CH3) 3

(BL-p2024)

  4 ml of a 10% aqueous solution of sodium iodide is added to a stirred suspension of 14.6 g (48.7 mmol) of BL-P2013 (5) in 200 ml of acetone and the mixture is brought at reflux in a steam bath. 14.8 ml (0.1 mol) of redistilled chloromethyl pivalate (bp 34 ° C. at 933 Pa) were added at reflux to the refluxing suspension at once. The mixture is stirred at reflux for 3 hours, then cooled to room temperature.

  room temperature (22 C). Filtration is

  The crystalline solids were washed 3 times with ml of acetone, after which the filtrates were combined and evaporated in vacuo below 22 ° C to an oil. The oil is then taken up in 500 ml of ethyl acetate and the solution is washed once with 200 ml.

  of water and once with saturated sodium sulphate,

  say that it is stirred with 2 g of discolored charcoal under cooling in an ice bath. After 20 minutes, the mixture is filtered through diatomaceous earth (Dicalite) by depression, after which the dicalite is washed 4 times with 100 ml of ethyl acetate. The filtrates are combined and concentrated at 22 ° C under

  to an oil. The oil is concentrated further

  around 22 C below 133 Pa to eliminate the

  much of the residual chloromethyl pivalate. The remaining oil is then triturated twice in 50 ml of n-pentane and left to stand during the weekend in a cold room (approximately 10 C) under a layer of

  n-pentane. The crystalline solid mass is then crushed

  resulting line into a solid powder under 40 ml of a 4: 1 mixture of ether and n-pentane. The product is collected by filtration, washed with a 1: 1 ether-pentane mixture and then with pentane, after which it is

  dry in the air. After drying under vacuum for 4 hours

  in the presence of P 2 O 5, 13.37 g of pivaloyloxymethyl 23-chloromethyl-2 "-methylpename-35-carboxylate sulfone (about 75% yield) was obtained.

at 93-95C.

  Analysis for C14H20ClNO7S Calc'd: C, 44.03; H, 5.27; N, 3.67%; Found: C, 44.11; H, 5.08; N, 3.85%

EXAMPLE 4.-

  Recrystallization of 2p-chloromethyl-2α sulfone

  potassium methylDéname-3a-carboxylate (BL-P2013) Water is added at a rate of 1 ml to the

  stirred with a mixture of 20 ml of n-butadiene

  and 1 g of BL-P2013 (5) in a separatory funnel until a yellow solution forms

  blade. The clear solution is filtered through filter paper.

  pleated, then the ampoule and the filter paper are washed with about 10 ml of a 9: 1 mixture of n-butanol and

  of water, after which the filtrates are combined and

  read with another 20 ml of n-butanol. We introduce the

  resultant solution in the round bottom flask of an

  rotary porator and evaporated under reduced pressure to about half of the initial volume. We are

  collect by filtration the snow white crystals of the

  it is washed 6 times with 10 ml of acetone and dried in air. This gives 810 mg. After 6 hours drying under vacuum on P205 below

  133 Pa, we obtain 800 mg (80% yield) of

  melted at 215 ° C with decomposition.

  Analysis for C 8 H 9 ClNO 5SK · 1 SO 8 K.I120 Calc'd: C, 29.67; H, 3.39; N, 4.63; C1, 10.94;

K.F.H20, 5.56%

  Found: C, 29.23; H, 3.38; N, 4.49; C1, 10.74

K.F.H20, 5.74%

  This recrystallization technique gives a crystalline monohydrate which is different from the compound

  starting material, which is essentially anhydrous.

EXAMPLE 5

  ## STR1 ## ## STR1 ##

-

  See Chemical Abstracts 27, 24271 and 22, 3828; outraged

GB 299064.

o C1S03CH2C1 (6 'S CH% 2Cl 3S CHCl 2

"CH3 NH

N C-OCH Cl

I00K 2

z NaI acetone | 0 W0 ES / CH 2 Cl "CH

O '-OCH2I

o8 8 +

H3C C N

  E I C + I. OCH3O309 (U.S. Patent 3,316,247) CH3I

\ / CHC0O-NH

I "3 It O.0 2H

0 -0

0o II 1 f.r.t v; U n s

C H CO _NHC

N1i2 0 C = 0 r_

- 110 I1

CH l I2 0C - _--- O

"- 0 0

/.-CH.C

H2

0 / -./, CH3

II

Z491071

  a) A solution of chloromethyl chlorosulfate (0.115 mmol) in 40 ml is added dropwise

  of dichloromethane, maintaining the temperature of

  at 30 ° C. to a solution of compound 2 (0.1

  1c) and potassium bicarbonate (0.3 mole) in 200 ml of a 1: 1 mixture of dichloromethane and water containing

  tetrabutylammonium hydrogen sulphate (0.01 mol).

  At the end of the addition, the mixture is stirred at room temperature.

  at room temperature for 30 minutes, then the organic phase is separated off and the aqueous phase is extracted with

  dichloromethane (50 ml). The organic phases are combined

  These are then dried over sodium sulfate and evaporated in vacuo to give a residue which is dissolved in 150 ml of ether. After addition of diatomaceous earth, the insolubles are separated by filtration and the filtrate is evaporated under vacuum to obtain the

compound 7.

  b) 1.6 g of bis-chloromethyl sulphate are added

  thylated to a suspension of the compound 5 (1.5 g) in

  dimethylformamide (12 ml) and the mixture is stirred

  45 minutes at room temperature. After di-

  The mixture is washed with ethyl acetate (50 ml), the mixture is washed with water and then with aqueous sodium bicarbonate, after which it is dried and evaporated under vacuum to give the mixture.

  compound 7 in the form of an oil.

  c) 7 mmol of triethylamine and mmol of chloroiodomethane are added to a solution of mmol of the compound in 7.5 ml of dimethylformamide, and the mixture is stirred for 4 hours at room temperature.

  ambient temperature. After dilution with 30 ml of

  ethyl acetate, the mixture is washed 3 times with 10 ml

  water, then with 5 ml of saturated aqueous sodium chloride

  after which the mixture was dried and evaporated in vacuo to give 7 as a

oil.

  d) 1.5 moles of chloroiodomethane are added to a mixture of 0.15 moles of compound 5, 0.15 moles of silver nitrate and 7.5 grams of silver oxide in 750 ml of acetonitrile . After stirring for 48 hours at room temperature, the silver salts are filtered off and the filtrate is evaporated to dryness in vacuo. The residue is dissolved in 200 ml of ethyl acetate and the solution is washed with sodium chloride.

  saturated aqueous solution, it is filtered, dried and

  pore under vacuum to obtain the compound 7.

  Compound 7 and other intermediate compounds are purified, as are the end products of

  the invention, if desired, by chromatogra-

  on Sephadex column LH20 using a

  chloroform: hexane 65:35 as eluent, for example

  ple, or by chromatography on a silica gel column

  this, for example Mallinckrodt CC-7, using

  hexane-ethyl acetate 3: 2, ethyl acetate-

  petroleum ether 8: 2, 7: 3, 1: 9 or 15:85, acetate of

  thyl-n-hexane 4: 6 or 3: 1, hexane-ethyl acetate 3: 1,

  1: 1 or 1: 4 or cyclohexane-ethyl acetate 1: 1.

  The thin-layer chromatography is useful too. "Sephadex" is epichlorohydrin,

  2- (diethylamino) ethyl ether hydrochloride

  De [[2- (diethylamino) ethyl] diethylammoniolethyl

  crosslinked trane (Merck Index, ninth edition, No. 7337).

  A solution of 0.2 mol of compound 7 and 0.3 mol of sodium iodide in 150 ml of acetone is stirred at room temperature for 18 hours. The resulting suspension is cooled to about 0.degree.

  it is adjusted to about pH 7.2 by addition, under stirring.

  saturated aqueous sodium bicarbonate. After

  bleaching by the gentle addition of sodium thiosulphate

  0.5 M aqueous dium, 150 ml of water drop

  drop to the agitated mixture to precipitate the

  The solid was collected by filtration and washed with 1: 1 acetone-water (2 x 20 ml), isopropanol (2 x 20 ml) and ether (2 x 20 ml). ml)

then we dry.

  Ampicillin is converted to compound 9 using methyl acetoacetate as described in U.S. Patent No. 3,316. 247. 0.5 mole of compound 8 is then added to a solution

  0.57 mol of the compound 9 in 1 liter of

  formamide. After stirring for 15 minutes at 5 ° C., the reaction mixture is poured into an ice-cold stirred mixture of 4 liters of ethyl acetate and 2 liters of saturated aqueous calcium chloride. The organic phase is separated, washed with calcium chloride

  Saturated aqueous solution (2 x 500 ml), filtered and

  pore up to about 1 liter under vacuum to obtain a concentrated solution of compound 10. 500 ml is added

  of water and 500 ml of n-butanol to this concentrate, then,

  4 N hydrochloric acid with stirring until removal of the radical protecting the function.

  amino, so as to obtain a solution of compound 11.

  At the end of the addition of acid, 1 liter of

  ther and 500 ml of water to the stirred mixture, after which

  separates the aqueous phase and extracts the organic phase

  than with 800 ml of water. The aqueous extracts are combined and washed with 1 liter of ether, and then added

  640 g of sodium chloride and 2 liters of dichloromethane

  after which the mixture is stirred for 15 minutes.

  your. The organic phase is separated off and the aqueous phase is extracted with 1 liter of dichloromethane, after

  organic extracts are combined with them and

  che on magnesium sulphate, then evaporate

  up to about 600 ml under reduced pressure to obtain

  hold a concentrated solution of the compound 11. By

  200 ml of 2-butanone are added to the concentrate and

  cold, the solid state is precipitated

  2p-chloromethyl-2 "-methylpenam-3c-carboxy-sulfone

  6 - [(R) -2-amino-2-phenylacetamido] -3,3-dimethyl-

  7-oxo-4-thia-l-azabicyclo [3.2.O] heptane-2--alkylcarbonyloxy-

  methyl (11) which is collected by filtration.

Z491071

EXAMPLE 6

  Sulfone of 2-chloromethyl-2α-methylpenam-3c-carboxy

  6-E (R) -2-amino-2-p-hydroxyphenylacetamido] -3,3-

  dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2-car-

bonyloxymethyl of formula:

S CH3

HO CH-C0 -NH C

NOT

MH2 C = 0

- 0b o e CH2C1 JIÈ uIICH3

2O- O0

II

  This compound is obtained by replacing the ampicillin used with

  in Example 5 with amoxicillin.

EXAMPLE 7

  a) CE / N-Bromosuccinimide O-azo-isobutyronitrile A 14 CHBr

12%

  As described in US Pat. No. 3,860,579, 50 g (0.375 moles) of recrystallized phthalide and 0.375 moles of recrystallized N-bromosuccinimide were refluxed for 4.5 hours in the presence of about 100 mg of a-azobutyronitrile in 1 liter of carbon tetrachloride. The mixture is cooled to about 15 ° C. and filtered to separate

  succinimide which is washed in turn with 100 ml of

  carbon trachloride which is separated by filtration. The solutions are mixed in carbon tetrachloride and concentrated in vacuo to about 150 ml to obtain solid 3-bromophthalide which is collected by

  filtration, which is washed with about 50 ml of tetrachloro-

  Carbon monoxide and air dried to obtain 54 g of the compound whose weight reduces to 50 g after recrystallization in boiling cyclohexane, the melting point being 84-86 Co b) o 0 CHBr

> CH2C1 0

H C-OK It 0o DMF tD4

v25 22 C.

o> o S st CH C1

2

j. 'CH3

NOT " *

C-O-CH BL-P2056

0 0 3 \ CI * dl

_

  1.7 g (8 mmol) of 3-bromophthalene are added.

  lide (12) to a partial solution and suspension

  of the compound 5 (BL-P2013, 2.3 g, 7.6

  1c) in 20ml of dimethylformamide (dried for 3 weeks on 3A molecular sieves) and the mixture is stirred for 4 hours at 22 ° C. The resulting mixture is poured into a mixture of 200 ml of ice water and 1 ml of water. iced ethyl acetate (rinsing the balloon

  with a little ethyl acetate), then stir the mixture

  lange. The organic phase is then separated and washed 7 times with 100 ml of ice water. The organic phase is washed again with aqueous sodium sulphate

  saturated and then cold-dried on sodium sulfate

  dium, filtered and evaporated to dryness in vacuo to give an oil as tracer residue, triturated twice in 25 ml of cyclohexane, twice in 25 ml of "Skellysolve B" (P.Eb 60-68 C, essentially from

  n-hexane) and 4 times in 25 ml of n-hexane, to obtain

  2.5 g of the compound 11 in the form of a solid

  than white after air drying. The product is then dried over P 2 O 5 under a pressure of less than 133 Pa to obtain 2.5 g of compound 13, mp 104 ° C.

  decomposition. The estimated purity is 85-95%.

  Analysis for C₁HH₁CClONNO7S Calculated: C, 50.61; H, 3.79; N, 3.77; Cl, 9.53% Found: C, 52.59; H, 4.67; N, 3.21; Cl, 7.73%

K.F.H20, 0.27

EXAMPLE 8

  2-Chloromethyl-20-methylpena-3a-carboxy sulfone

  pivaloyloxymethyl late mixture is stirred into a mixture of 1 g

  (3.1 mmol of 23-chloromethyl-2α-methylpiperazine sulfone)

hydrated potassium naphth-3α-carboxylate and 1 g of molecular sieve 3A to 15 ml of dimethylacetamide for 2 hours at 23 ° C. To this mixture is added 470 mg (3.1 mmol) of pivaloyloxymethyl chloride.

  and stirring continued for 18 hours. We

2 4 9 1071

  the molecular sieve and dilute the filtrate with

  ml of water and then extracted with ethyl acetate.

  The ethyl acetate extract is washed 9 times with water and then dried over anhydrous magnesium sulfate. The solvent is removed at 30 ° C. (2000 Pa) to

  obtain an oil that is chromatographed on

  Silicar CC-7 (methylene chloride 8, acetate)

  of ethyl 2) giving a spot whose Rf is 0.5.

  The residue is crystallized from heptane ("Skelly-

  solve B "') to obtain 100 mg of 20-chlorosulfone

  pivaloyloxy methyl-2α-methylpénam-3c-carboxylate

methyl melting at 94-95 C.

  Analysis Calculated: C, 44.03; H, 5.27; N, 3.67% Found: C, 44.20; H, 5.24; N, 3.63%

  The nuclear magnetic resonance spectrum and the

  infrared spectrum are compatible with the structure.

EXAMPLE 9

  Sulfone of 20-chloromethyl-2α-methylpenam-3α-carboxy

  Sodium salt. ## STR2 ## ## STR2 ## C3 N f. (1) (2) 0N Co2K (306) 2 Cs8H9ClNO5SNa

(306) (289.67)

  2N hydrochloric acid is added to a

  stirred solution of 500 mg BL-P2013 (potassium salt)

  sium) in 5 ml of water and 10 ml of ethyl acetate

  that the pH is 1 (we carry out the operation

  under intense agitation in the ice bath). We are saturated

  After mixing the sodium sulphate, separate the

  aqueous layer and dry the organic phase briefly

  In ice-water over sodium sulfate, after which it is collected by filtration and 50% sodium 2-ethylhexanoate in anhydrous n-butanol is added dropwise until the pH is wet. The product does not crystallize during the scraping of the container wall and the mixture is then concentrated in vacuo to an oil which is dissolved in ml of acetone, the scraping of the walls does not induce crystallisation and a supply of ether up to the point

  trouble does not induce crystallization either.

  The mixture is concentrated under vacuum in a rotary evaporator.

  to obtain an oil that is dissolved in

  ethyl acetate and to which is added a drop of water

  without scratching the crystals.

  The mixture is concentrated under vacuum and the residue is triturated.

  sidu with 5 ml of n-butanol to obtain 200 mg of

  amorphous white powder which is washed with ether, which is

  air and vacuum under P205 for 24 hours. Finally, 180 mg of sulphone of

  2β-chloromethyl-2α-methylpenam-3α-carboxylate of

  dium whose poorly defined decomposition point is

less than 1000 C.

  Analysis for C8H9ClNO5SNa Calc'd: C, 3355.10; H, 5.153; N, 4.89% TP found: C, 33.20; H, 3.69; N, 4.44; H20 according to K.F., 4.04%

EXAMPLE 10

  Sulfone of 25-chloromethyl-2-methylpname-3-carboxylate

of potassium (BL-P2013)

  150 g (1.25 moles) of hydrogen carbonate are added.

  sodium nate and 200 g of 10% palladium on barium sulfate to 10 liters of water and 272 g

  (0.565 moles) 6α-bromo-2β-chloromethyl-2-sulphone

  p-nitrobenzyl methylpenam-3-carboxylate in 5

  very much of ethyl acetate. We mix everything and we do

  hydrogenation at 40 C under a pressure of 101 kPa. After 5 hours, the absorption of hydrogen becomes very slow and 200 g of palladium are added to

  % on barium sulphate, after which the hyaluronide is

  until the hydrogen uptake

come imperceptible.

  The suspension is filtered through diatomaceous earth ("Celite"), the filter cake is washed.

  with water and the aqueous phase is washed with 3 liters of

  ethyl acetate. 3 liters of ethyl acetate are added to the aqueous solution and the pH of the mixture is brought to 1.5 by means of 150 ml of 12 N hydrochloric acid.

   C. Separate the organic phase and saturate the

  aqueous solution with sodium sulfate decahydra-

  after which the extraction is carried out twice with 1 liter of ethyl acetate. The extracts are combined and dried over magnesium sulfate. We separate

  drying agent and 260 ml of 2-ethylhexa are added.

  2N potassium notate in butanol at 0 C.

  After stirring for 2 hours at 0 ° C.,

  and at room temperature, the potassium 23-chloromethyl-2-methylpenam-3-carboxylate (BL-P2013) sulfone was dried under reduced pressure. 134.8 g are thus obtained

  (about 70% yield) of this compound.

EXAMPLE 11

  P-nitrobenzyl 6x-bromopenicillanate sulphoxide

0

* BrI '+ TEA + BrCH2 NO

N CO2H (101) (22

(216) o2H

3 0 (296)

Or Br / -

O 0 CH NO

2 2 2

  (44 g (0.148 mole) of sulfoxide is added

  6a-bromopenicillanic acid, then 20.5 ml (0.148 mm).

  triethylamine) and 38.2 g (0.177 mole) of bromide

  p-nitrobenzyl to 200 ml of N, N-dimethylacetamide.

  The mixture is stirred at 22 ° C. for 20 hours. The reaction mixture is poured into 1 liter of water which is extracted 3 times with 300 ml of methylene chloride.

  methylene and washed with 200 ml of aqueous solution

  to 5% sodium bicarbonate, after which they are

  on sodium sulphate at 5 ° C. for 30 minutes.

  The solution is filtered and evaporated in vacuo to give a residue. The residue is diluted with ether and the solid is collected by filtration to obtain, after drying, 54 g of 6O-bromopenicillanate sulfoxide.

  p-nitrobenzyl. The yield is 85%.

  The nuclear magnetic resonance spectrum is

with the assigned structure.

  The yield attained at this stage is the same as for the esterification of the potassium salt. The advantage is that it

  It is not necessary to prepare the potassium salt.

  (This operating phase is done with a yield of

at 90%).

EXAMPLE 12

  Prenaration of p-nitrobenzyl 6α-bromopenicillanate sulfoxide 873.0 g (2.95 moles) of sulfoxide were added

  of 6a-bromopenicillanic acid (S) at 4.375 liters of N, N-

  dimethylacetamide, then 293 g (2.95 moles) of triethylamine and then 764 g (3.54 moles) of p-nitrobenzyl bromide were added with stirring and maintaining the temperature of the system below C. The mixture is stirred at room temperature for 5 hours.

  and let him rest until the next day.

  The reaction mixture is poured into 20

  water extracted 3 times with 7 liters of chlorine

  methylene chloride. The organic extracts are combined and washed 5 times with 7 liters of water, then once

  with 7 liters of a 5% aqueous solution of bicarbonate

  sodium nate, after which they are dried over sulphate

anhydrous magnesium.

  The magnesium sulfate is filtered off and the solution is evaporated to give a crystalline residue, 4 liters of diethyl ether are added to the residue and the crystals are collected to give, after drying at room temperature, 1171 g (yield:

  92%) of 6α-bromopenicillanate sulfoxide from p =

  trobenzyle. On analysis, the compound shows a bromine content of 18.48% (calculated value 18.53%) and a

  rotary DNA +162 (0.25% in methanol).

EXAMPLE 13

  Preoaration of 6a-bromo-2β-chloromethyl sulfone

  P-nitrobenzyl 2-methylpenamino-3-carboxylate

  364.6 g (0.812 moles) of 6α-bromine are added.

  2β-Chloromethyl-2-methylpenam-3-carboxylate of p-ni-

  trobenzyl to 16 liters of acetic acid. The resulting solution is stirred at room temperature and a solution of 282 g (1.78 moles) of potassium permanganate in the course of 3 hours is added dropwise thereto.

  26 liters of water. The mixture is then stirred at room temperature.

  room temperature for 1 hour and add

  37% hydrogen peroxide until it reaches

  nish a colorless solution. 30 liters of water are then added, after which the mixture is stirred for 1 hour at room temperature and the crystalline precipitate is separated off and washed 5 times with 5 liters of water and twice with 2 liters of ethanol. before drying it under

vacuum at room temperature.

  297 g (yield of 76%) of the compound having a rotational power aD of +75.9 (0.5%) are thus obtained.

in methylene chloride).

EXFViPLE 14l.-

  Preaction of the free acid BL-P2015 and CH2CI cICY2Cl 113 F04 J c1 0'c2H

  800 mg (2.61 mmol) of potassium salt are added.

  of BL-P2013 to a mixture of 25 ml of acetate acetate.

  thyle and 10 ml of water. After complete dissolution of

  solid, the mixture is added dropwise under

  stirring, 50% aqueous phosphoric acid

  only at the end of precipitation from the layer

  aqueous. The phase formed by acetate acetate is separated

  thyle, then washed with a saturated solution of

  sodium chloride and dried over magnesium sulphate

  anhydrous gnesium. The desiccant is separated by

  and washed with 10 ml of ethyl acetate. We

  mix the washings with the initial filtrate.

  "Skellysolve B" is then added to the ethyl acetate until the cloud point (approximately 10 ml). 500 mg activated charcoal ("Darko KB") is added to the mixture and filtered. The filtrate is diluted with 15 ml of "Skellysolve B" and then seeded with crystals of BL-P2013 as free acid. After about 3 hours at room temperature, the free acid crystalline precipitate is collected and dried under vacuum for 15 minutes on P2O5 to obtain 323 mg (46% yield) of the melting compound above 100 ° C.

with slow decomposition.

  Analysis for C8HloClNO5S Calc'd: C, 35.89; H, 3.77; N, 5.23; Cl, 13.25% Found: C, 35.88; H, 3.91; N, 5.41; Cl, 13.52% This compound is unstable in 7 days

conservation at 23 C.

EXAMPLE 15

  6 "-bromopenicillanic acid sulfoxide" 0

J

(C6H5dH25 1cH2) 2-> C0

2

PM 800.64PM 296 14

  300 g (0.75 mole) of N, N'-salt are added.

  6-bromopenicillanic acid dibenzylethylenediamine to 3 liters of methylene chloride and the suspension is cooled to 5 ° C. 130 ml of concentrated hydrochloric acid are then added dropwise and with vigorous stirring over 15 minutes. The suspension is stirred at 5 ° C.

  during 2 hours. It is then filtered on the ground

  diatom ("Celite"), then wash the cake

  filtration 3 times with 250 ml of methylene chloride.

  The solutions are combined in methylene chloride and washed twice with 500 ml of water and then dried over sodium sulfate for 15 minutes. The sodium sulfate is filtered off and the filtrate is evaporated under reduced pressure until

about 750 ml.

  The solution is cooled to 5 ° C. and with vigorous stirring, 130 ml of 40% peracetic acid are added drop by drop while maintaining the temperature between 12 ° C. and 12 ° C. The addition is strongly exothermic. At the end of the addition, the suspension is stirred at 5 ° C. for 2 hours, then the product is isolated by filtration and

  Wash it with 5 ml of cold water (5 C) and 100 ml of chlorine

  cold methylene chloride (5 C). This gives 126 g

  (yield 57%) of 6a-bromopenic acid sulfoxide

  cilanic melting at 129 C. The infrared spectrum and

  the nuclear magnetic resonance spectrum are

  patible with the structure of the desired product.

  Analysis for C8H10BrNO4S Calc'd: C, 32.44; H, 5.40; N, 4.73% Found: C, 32.30; H, 3.35; N, 4.71%

H20, 2.18

  Potassium 6a-bromopenicillanate sulforoxide

O O

Br + Br, s

"C0 H0

02H 2K

PM 334.24

  126 g (0.43 mol) of sulfoxide of

  6-bromopenicillanic acid and 162 ml of 2-ethylhexane

  potassium notate at 50% by weight in butanol to 3 liters of acetone. After stirring for 1 hour at 22 ° C., the product is collected by filtration, washed twice with 250 ml of acetone and dried. We obtain

  thus 127 g (90% yield) of 6-bromine sulfoxide.

  potassium mopenicillanate melting at 185 C. The

  infrared and the magnetic resonance spectrum

  are compatible with the desired structure.

  Analysis for C8H9BrKNOOS Calc'd: C, 28155; H, 2.71; N, 4.19% Found: C, 29.03; H, 2.78; N, 4.04% Pnitrobenzyl 6a-bromopenicillanate sulfoxide

0 0

+ Br +

F AC 0K Y C 2 CH 2 N 2

PM 431.28

  145 g (0.43 mol) of sulfoxide are added

  Potassium 6a-bromopenicillanate to 1 liter of N, N-

  dimethylacetamide, while stirring 115 g (0.53

  1a) of p-nitrobenzyl bromide at 22 ° C. The mixture is stirred at 22 ° C. for 20 hours. The reaction mixture is poured into 3 liters

  of water extracted 3 times with 1500 ml of acetate acetate.

  Thyle. The ethyl acetate extracts are combined and washed twice with 500 ml of aqueous solution

  5% sodium bicarbonate and then dried

  30 minutes on sodium sulphate. The sodium sulfate is filtered off and the filtrate is evaporated under reduced pressure to give a residue to which 1 liter of diethyl ether is added to crystallize the product. The crystals are collected by

  filtered, washed twice with 100 ml of diethyl ether.

  ethyl ether and dried to give 162 g (87%) of p-nitrobenzylabromopenicillanate sulfoxide melting at 1110 C. The infrared spectrum and the nuclear magnetic resonance spectrum are compatible with

the desired structure.

  Analysis for C₁HH₁BrBrN₂O6S Calculated: C, 41.78; H, 3.51; N, 6.50 Found: C, 41.66; H, 3.45; N, 6.85; H20 according to K.F. 0.69 6a-Bromo-2-p-chloromethyl-2-methylp-N-dicarboxylic acid-3-carboxylate Br S-CH 2 Cl Br 1

C NO2 NO2

  35 μM EM Li 431.71 70 g (0.16 moles) of p-nitrobenzylphbrominopenicillanate sulfoxide are added, followed by 21.2 ml.

  (0.10 mole) benzoyl chloride and 21.8 ml (0.19

  1c) of quinoline to 1 liter of p-dioxanes. The reaction mixture is refluxed for 4 hours, then cooled to 220 ° C., poured into 2500 ml of water and extracted 3 times with 800 ml of acetate.

  ethyl. The extracts are combined with ethyl acetate

  and washed with 300 ml of 5% aqueous sodium bicarbonate solution, 300 ml of 5% aqueous phosphoric acid and 300 ml of water. The solution is dried in ethyl acetate for 30 minutes on sodium sulphate and the sodium sulphate is separated off.

  filtration. The filtrate is evaporated under reduced pressure.

  to obtain a residue which is redissolved in 1 liter of ethyl acetate, after which the solution is evaporated.

  again under reduced pressure to obtain a residue.

  1 liter of diethyl ether is added to the residue and the product is collected by filtration to obtain 41 g.

  (57% yield) of 6m-bromo-23-chloromethyl-2-methyl-

  p-nitrobenzyl penam-3-carboxylate melting at 1320 C.

  The infrared spectrum and the magnetic resonance spectrum

  nuclear technology are compatible with the structure

  Siree. Analysis for C15H14BrClN2O5S Calc'd: C, 40.06; H, 3.14; N, 6.23% Found: C, 40.62; H, 3.11; N, 6.13% I1

  6m-bromo-23-chloromethyl-2-methylthiomethyl sulfoxide

  P-Nitrobenzyl-3-carboxylate ## STR2 ##

NO2 C02CH2

NO2

PM 465.71

  51 g (0.11 mol) of 6α-bromo-23-

  p-nitrochloromethyl-2-methylpenam-3-carboxylate

  benzyl, followed by 23 g (0.12 mole) of m-chloroperoxy acid

  benzoic acid to 1200 ml of methylene chloride. The solution is stirred at 22 ° C. for 2 hours and evaporated under reduced pressure to obtain a moist residue. We

  This is stirred with 4 liters of diethyl ether

  hour, and then allowed to stand at 10 ° C for hours. The product is collected by filtration

  crystallized, washed twice with 200 ml of diethyl ether

  ethyl acetate and dried to obtain 39 g

  75%) of p-nitrobenzyl 6-bromo-23-chloromethyl-2-methylphenam-3-carboxylate sulfoxide melting at

  C. The infrared spectrum and the resonant spectrum

  this nuclear magnetic are compatible with the structure

desired form.

  Analysis for C₁HH₁BrBrCN₂OSS Calculated: C, 38.69; H, 3.03; N, 6.07% Found: C, 38.98; H, 3.04; N, 5.84%

H20, 0.35

  Sulfone of 20-chloromethyl-2-methylpenam-3-carboxy

potassium late (BL-P2013)

0 0

  --0 By. C2 H2C1 - j "" CH3 o0Co 'N02 -CO K

BL-P2013

PM 305.77

  8 g of 30% palladium on "Celite" and 16 g (0.19 mole) of 600 ml sodium bicarbonate are added.

  of water. 32 g (0.069 mol) of sulphate are then

  6a-bromo-2p-chloromethyl-2-methylpena-3-oxide

  p-nitrobenzyl carboxylate in 400 ml of acetate

  of ethyl and the solution is added to the aqueous dispersion

  is. The mixture is hydrogenated in a Parr apparatus at 345 kPa at 220 ° C for 4 hours. The suspension is filtered through a thin layer of "Celite" in a sintered glass filter, and then the filter cake is washed twice with 50 ml of water, after which the layer is separated.

  aqueous mixture of filtrates and liquors from the

  husbandry. The aqueous layer is washed with 200 ml of diethyl ether.

  ethyl ether, then cooled to 5 ° C. and with stirring, a solution of 12 g (0.076 mole) of potassium permanganate in ml of water is added dropwise over 30 minutes, keeping the pHI between 7.5 and 8.0 per

  addition of phosphoric acid at 40% o. When the

  pink diet persists for 5 minutes, one interrupts

  the addition of the potassium permanganate solution.

  The reaction mixture is stirred with a small amount of

  titer (about 50 mg) of sodium bisulfite for 30 minutes, and then the suspension is filtered through a layer of "Celite". The filter cake is washed twice with

  ml of water. The filters and liquors of the-

  500 ml of ethyl acetate and,

  with stirring, the pH is adjusted to 1.5 by means of a-

  Hydrochloric acid 2 N. The layers are separated and the aqueous layer of sodium sulfate saturated. We

* the extract back with 2 times 400 ml of acetate acetate

  thyle, then combine the extracts into the acetate

  of ethyl and dried over sodium sulfate

  30 minutes at 5 ° C. Sodium sulfate is filtered off and the filtrate is evaporated under reduced pressure to give a residue. This residue is dissolved in 160 ml of acetone and 160 ml of diethyl ether,

  then a 50% by weight solution of 2-ethyl-

  potassium hexanoate in n-butanol to neutral

  Trace the solution with moist pH paper. The potassium salt of BL-P2013 is crystallized,

  collected by filtration, washed with diethyl ether

  and dry it. This gives 16 g (

  76%) of 2p-chloromethyl-2-methyl sulphone

  potassium penam-3-carboxylate melting at 202 C.

  The infrared spectrum and the resonance spectrum

  Nuclear genetics are compatible with the desired structure. Analysis for C8H9ClKNO5S Calc'd: C, 31.42; H, 2.97; N, 4.58% Found: C, 31.18; H, 2.98; N, 4.51%

H20, 0.93

EXAMPLE 16.-

  Divaloyloxymethyl 23-chloromethyl-2-methylpename-3-carboxylate sulfone (BL-P202.4)

0 - 0 0

S CH C H2C1

3H35CCH3

o "2 Co2CH CCcCH3).

BL-p2024

PM 381 83

  4 ml of a 10% aqueous solution of sodium iodide is added to a stirred suspension of 14.6 g

  (48.7 mmol) of 2-chloromethyl-2-methylpiperazine sulfone

  potassium name-3-carboxylate (BL-P2013) in 200 ml of acetone and the mixture is refluxed in a molten bath. To the suspension boiling under reflux is added

  14.8 ml (0.1 mole) of chloromethyl pivalate redissolved

  (P.Eb.34 C at 933 Pa) at once. We shake

  the mixture is refluxed for 3 hours and then

  chills to room temperature (22 C). The crystalline solids were collected by filtration, washed 3 times with 30 ml of acetone, after which the filtrate mixture was evaporated under reduced pressure below 22 ° C to obtain an oil. The oil is taken up in 500 ml of ethyl acetate and the solution is washed once.

  with 200 ml of water and once with 200 ml of solution

  sodium sulphate. The solution is dried

  following briefly on sodium sulphate, while at the same time

  with 2 g of bleaching charcoal under cooling

  in the ice bath. After 20 minutes, the mixture is filtered through a layer of "Celite" and the filter cake is washed with 4 times 100 ml of ethyl acetate. The filtrate mixture is concentrated at 22 ° C. under reduced pressure to obtain an oil. The oil is further concentrated at about 22 ° C below 133: Pa to remove most of the residual chloromethyl pivalate. The residual oil is then triturated twice

  with 50 ml of n-pentane, after which it is left

  during the weekend at about 100 C under a layer of n-pentane. The resulting solid crystalline mass was crushed to a powder under 40 ml of a 4: 1 mixture of diethyl ether and n-pentane. The product was collected by filtration, washed with a 1: 1 mixture.

  of diethyl ether and n-pentane, then with n-penta

  After drying it with airo After drying under high vacuum for 4 hours on P205, we obtain

  13.37 g (about 75% yield) of 20-chloro sulfone

  pivaloyloxy-2-methyl-pivalen-3-carboxylate

  methyl (BL-P2024) melting at 93-95 C.

  Purification of BL-P2024 Approximately 3 g of crude BL-P2024 (obtained as described above) are dissolved in 5 ml of ethyl acetate, and the solution is then poured onto a column of 4.5 cm × cm of gel. of silica (Mallinckrodt CC7) which is eluted with a 4: 1 mixture by volume of methylene chloride

  and ethyl acetate. The fractions

  a single spot with a Rf of 0.84 (MLC on eluted silica gel plates

  with a 4: 1 mixture of methylene chloride and

  ethyl acetate, detection by I2) and concentrated under reduced pressure to give 1.38 g of a crystalline solid. A fraction of 900 mg of this solid is dissolved in 5 ml of ethyl acetate and the solution is filtered and then diluted to about the cloud point with petroleum ether ("Skellysolve B"). after which it is stored at room temperature for 3 days. The resulting crystals were collected by filtration and washed with petroleum ether and dried to give 560 mg of

  Purified BL-P2024 melting at 100-101 C.

Analysis for C14H20ClN07S

  Calculated: C, 44.03; H, 5.27; N, 3.67% -

  Found: C, 44.11; H, 5.08; N, 3.85%

EXAMPLE 17

  Preparation of the ammonium salt of BL-P2013 1. A solution of 250 mg of the free acid form of BL-P2013 in 20 ml of a 1: 1 mixture is filtered into

  volume of acetone and methanol to obtain a solution

crystal clear.

  2. An anhydrous ammonium solution is prepared by adding 1 ml of ammonium hydroxide (30%,

  laboratory work) to 10 ml of a 1: 1 mixture

  lume of acetone and methanol, then adding to this

  solution 1 g of anhydrous magnesium sulphate under stirring

  model, after which the mixture is filtered through

  filter paper, the filtrate being hereinafter referred to

anhydrous ammonium ".

  3. Gradually add and carefully mix

  approximately 2 ml of the "anhydrous ammonium solution"

to the filtrate obtained in 1.

  4. 100 ml of diethyl ether are mixed with the mixture obtained in 3 to precipitate the salt.

of ammonium of BL-P2013.

  5. The white ammonium salt is isolated from the soil

  and washed twice with 50 ml of diethyl ether.

than each time.

  6. The resulting powder is dried at 35 ° C for

empty to the next day.

  7. Analysis Calculated: C, 33.7; H, 4.6; N, 9.8% Found: C, 33.66; H, 4.63; N, 10.12%

Anhydrous according to K.F.

  Microscopic aspect: crystalline substance.

EXEMPIE 18.-

  Preparation of sodium salt of BL-P2013 non-hygroscopic

  1. 50 ng of the free acid form of BL-P2013 are dissolved in 4 ml of a 1: 1 by volume mixture of acetone and methanol. The mixture is filtered to obtain a

clear solution.

  2. A solution of sodium 2-ethylhexanoate is prepared by dissolving 40 mg of sodium 2-ethylhexanoate in 10 ml of a 1: 1 mixture by volume of acetone

and methanol.

  3. The 10 ml solution obtained in 2 is added to the filtrate obtained in 1 and the mixture is thoroughly mixed.

  4. 10 ml of diethyl ether are mixed with

  lange obtained in 3 to precipitate the salt of

dium BL-P2013.

  5. The white salt is placed in diethyl ether for 1 to 2 hours and then separated from the

  solvent and washed 3 times with 5 ml of diethyl ether.

every time.

  6. The resulting powder is dried at 30 ° C for

empty to the next day.

EXEIMPLE 19.-

Recrystallization of BL-P2013

0 O O 'O

  CH2Cl recrystallization CHL | D H2O-acetone r -., CK2 O 02K.H2

  400 mg of BL-P2013 are dissolved in a

  minimum amount of a 1: 1 mixture by volume of acetone and water and the solution is diluted with 10 ml of acetone,

  then filter it out and dilute it again with

  ketone to about 25 ml, after which the container wall is scraped and after 30 minutes

  by filtration the crystalline hydrate which is carefully washed

  acetone, then air-dried and then under vacuum below 133 Pa overnight. We

gets 280 mg of the compound.

  Analysis for C 8 H 9 ClN 4 O.H 2 O Calc'd: C, 29.67; H, 3.39; N, 4.63; C1, 10, 94;

H20, 5.55%

  Found: C, 29.32; H, 3.32; N, 4.44; Cl, 11.31;

H20, 5.90%

3o

EXAMPLE 20

  N-N'-dibenzylethylenediamine salt of BL-P2013 BL-P2013 + 1/2 N, N'-dibenzylethylenediamine diacetate recrystallization CH2C1 acetone-ether CH3 and C 02H. C6H5CH2NH-CH2

2

  306 mg (1 mmol) of BL-P2013 are dissolved in 7 ml of water and the solution is added to a solution of

  mg (0.5 mmol) of N, N'-dibenzylethyl diacetate

  linediamine in 7 ml of water. The mixture is stirred in which the salt crystallizes and after about 10 to 15 minutes of stirring, the salt is collected by filtration, and then

  it is air dried to obtain 300 mg of N, N'-

  dibenzylethylenediamine of BL-P2013. The salt is recrystallized by dissolving it in about 10 ml of boiling acetone and diluting the solution with ether to the cloud point. This gives 260 mg of

  compound dried in air and dried under vacuum.

  Analysis Calculated: C, 51.69; H, 5.42; N, 7.53; Cl, 9.55% found: C, 49.359; H, 5.49; N, 7.05; Cl, 8.96% H20 according to K.F., 1.23

EXAMPLE 21

  Chloromethyl ester of BL-P2015 ## STR5 ## wherein R 2 is CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2

O "COOK + 3 KHC03

(305/7)

CH2C12

H20 0 0 i H '

C-OCH 2 C

o

(316.17)

7

  A solution of

  9.5 g (57.5 mmol) of ClCH 2 -O-SO 2 Cl in 40 ml of chlorine

  methylene chloride to a vigorously stirred mixture of 25 g (50 mmol) of BLP2013 (2), 15 g (0.15 mole) of KHCO3 and 1.7 g (5 mmol) of tetrabutylammonium hydrogen sulfate (Company Aldrich Chem Co.) in a

  mixture of 50 ml of water and 50 ml of

  thylène. The temperature rises to 26 ° C and upon completion of the addition (which takes about 15 minutes), the mixture is stirred for a further 30 minutes. Since the product crystallizes, an additional 400 ml of methylene chloride is added to obtain

  a solution. The layer formed by chlorine is separated

  methylene chloride and mixed with 50 ml of a

  After washing with methylene chloride, the mixture is dried by stirring with magnesium sulfate and 2 g of bleaching charcoal ("Darco KB") is added. After about 50 minutes, the mixture is filtered, concentrated to about 50 ml and 150 ml of isopropanol are added. We hunt the rest of the

  methylene chloride under reduced pressure. We are

  collected by filtration the resulting crystalline precipitate

  as long as it is thoroughly washed with isopropanol and air dried. After vacuum drying below

  133 Pa, 8.5 g of 2-chloromethyl sulfone are obtained.

  Chloromethyl 2-methylpenam-3-carboxylate (7)

  at 116 C (with decomposition and darkening above

beyond 100 C).

  Analysis for C9H11Cl2NO5S Calc'd: C, 34.18; H, 3.51; N, 4.43; C, 22.43% Found: C, 34.16; H, 3.45; N, 4.47; Cl, 22.46% H20 according to K.F., 0.33

  The purity estimated is 90 to 95%.

  Iodomethyl Ester of BL-P2013 O C C1201 CH Cl S Na I

CH, CH3

// CH2 / 1

  ## STR2 ## 3 g (20 mmol) of sodium iodide are added.

  to a stirred mixture of 5 g (15.9 mmol) of chlorinated ester

  romethyl BL-P2013 (7) in 25 ml of acetone. The resulting suspension is stirred for 17 hours and then cooled to about 0 ° C. 2 drops of saturated aqueous potassium bicarbonate solution are added and the mixture is then slowly diluted.

  with water in 10 minutes to 50 ml.

  The suspension suddenly turns from yellow to gray and purple, then to black, and the crystals are immediately collected by filtration and washed with a cold mixture of acetone and water, 3 times ml of isopropanol. , diethyl ether and finally n-pentane, before drying in air to obtain 5.55 g (91% yield) of the iodomethyl ester of

  BL-P2015 (8) melting at 118-119 C with decomposition.

  The estimated purity is about 90%.

  Sulfone of 2-chloromethyl-2m-methylpena-35-carboxy

  6 - [(R) -2-amino-2-phenylacetamido] -3,3-dimethyl-

  7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2--alkylcarbonyloxy-

methyl (11) N il I OCH3

(Ampicillin Dane salt

born; see U.S. Patent No. 3,316,247)

CH-CO -NH

I N

H3C-CN -

H C /! 1H o0 o

O0 1

z, SC H2C1

JCH3 I '2

He H + H3C OC3 C0E

* ILI

Q C0t

C H -CO 0CN

lO-O

* 11 0 -O

0. - ** I-I0

- - '/, - eCHICl CH2 0Jj "CH3 -o

  4.08 g (10 mmol) of iodinated ester are added.

  of BL-P2013 to a stirred and ice-bathed mixture of 5.46 g (10 mmol) of Dane salt.

  of ampicillin above 9 (solvated by 1 molecule of

  isopropanol) in 60 ml of acetone and the solution is stirred

  almost limpid result for 5 hours

  pulling the ice bath after 30 minutes. Hunting

  then most of the acetone under vacuum

  rotary porator, then the resulting concentrated solution is dissolved in 200 ml of cold ethyl acetate, after which this solution is washed twice with 50 ml of ice water and twice with 100 ml of saturated aqueous sodium sulfate. The ethyl acetate solution is dried over sodium sulphate, filtered and the mixture is removed.

  most of the ethyl acetate under vacuum at

  rotary evaporator. The residue is triturated twice with 200 ml of dry diethyl ether and the resulting solids are collected by filtration to give 5.5 g of the compound as a pink powder. We incorporate this

  powder to a stirred mixture of 50 ml of water, 50 ml of

  butanol and 20 ml of ethyl acetate to which 6 N hydrochloric acid is added dropwise until the pH of 2.5.0n is added occasionally 1 or 2gocttes hydrochloric acid to maintain the pH at 2.2-2.5 for 45 minutes. When the pH ceases to rise, 100 ml of diethyl ether are added to the mixture with good stirring. The aqueous phase is separated and mixed with a second 25 ml aqueous extract of the organic phase. The aqueous solution is extracted once with 50 ml of diethyl ether and the ethereal extract is discarded. The aqueous layer is then stirred

  under a layer of 100 ml of 2-butanone (methylethyl-

  ketone), while sodium sulfate is added to saturate the aqueous layer. We separate the layer of

  2-butanone, dried for 30 minutes on sulphate

  Sodium salt in an ice bath, filtered and concentrated under vacuum to near dryness. The residual oil is triturated to a solid under n-butanol and then

  the solid is carefully washed with ether and then

  pentane, after which it is first dried in air, then

  under vacuum on P205 below 133 Pa to obtain

  1.6 g of 2p-chloromethyl-2-methylpena-

  6 - [(R) -2-amino-2-phenylacetamido] 3m-carboxylate

  3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2

  carbonylmethyl in crude form. The spectrum below

  red and the nuclear magnetic resonance spectrum are compatible with the structure of the compound (11) which is however not of high purity. It is estimated that this solid contains at least 40% by weight and can be

  up to 80% by weight of 2p-chloromethyl-2α sulfone

  6 '- [(R) -2-amino-2-phenyl) methylpena-3 "-carboxylate

  acetamido] -3,3-dimethyl-7-oxo-4-thia-l-azabicyclo

  [3.2.0] heptane-2-carbonyloxyméthyleo

EXAMPLE 22

  Improved synthetic route of BL-P2013 This procedure simplifies the preparation

  BL-P2013 by removing the catalytic reduction ex-

cut previously. Stage 1 Br

+ CH2CC13 + Dicyclohexyl-

O COOH. "Ocarbodiimide

0 --- N OH

COOH Pyridine CHC12

X

-OC-C C

  o (See page 633 of Cephalosporins and Penicillins

  Edwin H. Flynn, Academic Press, New York, 1972).

  30 g (0.1 mole) of sulfoxide of

  6a-bromopenicillanic acid (1) in 1000 ml of chlorine

  Dry methylene is then added to the solution 16.2 ml (0.2 mol) of pyridine and 29.8 g (0.2 mol) of trichloroethanol. 20 g (0.1 mol) of dicyclohexylcarbodiimide are then added and the mixture is stirred at 22 ° C. for 16 hours. Dicyclohexylurea begins

  to precipitate and finally removed by filtration.

  The filtrate is washed with 200 ml of sodium bicarbonate.

  5% aqueous dium, 200 ml 10% phosphoric acid and 100 ml saturated aqueous sodium sulfate. Dry

  the organic phase at 50 ° C. for 30 minutes on sulfur.

  sodium fate which is separated by filtration, after which the organic phase is evaporated to an oil. We add

  diethyl ether and, by scraping,

  to size product 2 (obtained in quantity of 27 g,

with a yield of 27%).

  Stage 2 2 Ol 10 CCl quinoline Br C 2 Cl N C-OCH-CCl 3

0

  Compound 2 (26.5 g, 62 mmol) was dissolved in 500 ml of p-dioxane and 8.5 ml (78 mmol) of benzoyl chloride and 8.75 ml (78 mmol) of quinoline were added to the solution. The solution was refluxed for 4 hours, then poured into 100 ml of water, after which the product was extracted 3 times with 300 ml of ethyl acetate. The ethyl acetate extracts are mixed, washed sequentially with 200 ml of 5% aqueous sodium bicarbonate, 5% phosphoric acid and 200 ml of saturated aqueous sodium sulfate. dried over sodium sulfate at 5 ° C. for 30 minutes and evaporated to an oil (3) which is used as is for

the next reaction.

  Stage 3 3 + KMnO4 + H22 in glacial acetic acid \ o B \\ A CH Cl

O 2

10 X <CH3 4

O C-OCH2CC13

o

  The compound 3 obtained at the pre-stage stage is dissolved

  in 1000 ml of glacial acetic acid and

  stirring at 22 °, a solution is added dropwise.

  saturated aqueous solution of potassium permanganate

  than a persistent pink color (that is, a drop on a piece of filter paper gives a

  pink spot). Under cooling, then add

  hydrogen peroxide at 30% o until

  a clear solution containing a little bit of pre-

  white precipitate. The solution is poured into 2500 ml of water

  and the product 4 is extracted 3 times with 500 ml of acetaminophenic acid.

  te of ethyl. The extract is washed in ethyl acetate

  with 5% aqueous sodium bicarbonate until neutral.

  trinity (that is to say until the end of effervescence),

* then it is dried over sodium sulphate and evaporated

  pore to obtain the compound 4 as a residue.

  The residue is left to stand at 100C for 1 day, then triturated with "Skellysolve B" to obtain 9.1 g of the solid 4. The yield is 28% of the value.

  theory in stages 2 and 3 together.

  q 6 Stage 4 4 + (D Zn in acetic acid + 0 2-ethylhexanoate potassium o /

\\ // 2-

iI 5 (BL-P2013)

O COOK

  (See U.S. Patent No. 4,164,497) 3.75 g of zinc dust are dispersed in 1 ml of glacial acetic acid and the dispersion is cooled to 50 ° C. To this mixture is added a solution of

  3 g of the compound 4 (5.7 mmol) in 15 ml of dimethyl

  formamide and the resulting suspension is stirred at 5 ° C.

for 2 hours and 30 minutes.

  The zinc is filtered off and the pale yellow solution is poured into 80 ml of 5% aqueous hydrochloric acid. The mixture is extracted 3 times with 25 ml

  of ethyl acetate. The extracts are mixed in the

  ethyl acetate and, in turn, extracted with 3 times 20 ml of 5% aqueous sodium bicarbonate

  retaining the ethyl acetate phase after the separation

  tion. The bicarbonate extracts are combined, covered with a layer of ethyl acetate, brought to pH 1.5 by addition of 2N hydrochloric acid and saturated with sodium sulfate. The ethyl acetate phase is separated and the aqueous phase is extracted twice.

with 30 ml of ethyl acetate.

  All the extracts are mixed in ethyl acetate, dried over sodium sulphate and

  evaporate to an oil (which is the acid form

  of BL-P2013) dissolved in about 20 ml of

  ketone solution to which 20 ml of diethyl ether

  ethyl. Potassium 2-ethylhexanoate is then added.

  50% potassium in dry n-butanol until neutral. Product 5 (BLP2013) is deposited as crystals. After

  minutes of stirring at 22 C, the crystals are collected

  filtration rate to obtain 650 mg of compound 5

(37% yield).

  50 mg of compound 5 are dissolved in 0.5 ml

  20 mg of N, N'-dibenzyl diacetate are added

  ethylene diamine. The dibenzylethylenediamine salt of

  The compound is deposited in crystals which are collected by

  then washed with water and dried on

  P205 under vacuum to obtain the 20-chloromethyl sulfone

  N, N'-dibenzylethyl-2-methyl-phenyl-3-carboxylate

  Linediamine (N, N'-dibenzylethylenediamine salt)

free acid).

  Another 450 mg sample of compound 5 was dissolved in 3 ml of water and 270 mg of N, N'-dibenzylethylenediamine diacetate in 2 ml of water was added to the solution. By scratching, we crystallize

  430 mg of the N, N'-dibenzylethylenediamine salt of

  5. Recrystallization from 100 ml acetone

  boiling, isolate 270 mg of the compound.

EXAMPLE 23

  20-Chloromethyl-2c-methylpena-3a-carboxy sulphone

  6 - [(R) -2-amino-2-p-hydroxyphenylacetamido] late

  3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] heptane

  2-carbonyloxymethyl of formula: ## STR2 ##

NH2 "--0

2O

154 0

0 0 o o s CH2Cl JjCH, C2 c3

ID

  This compound is prepared by replacing ampicillin in

  the method of Example 21 with the corresponding Dane salt

laying down of amoxicillin.

Biological data

  The compound of Example 1 or compound 5

  in the formula: o c1 CH2C1 "'CH N 0K

  is hereinafter called BL-P2013 compound.

  Although it is at best a very weak antibacterial agent, BL-P2013 inhibits 3-lactamases and protects ceforanide and amoxicillin against in vitro and in vivo destruction by P-producing bacteria. -Lactamase when combined with these two agents.

T A B L E A U I

  Antibacterial activity of the new sulfone.

  CIM (/ ug / ml) Organism B gi BL-P2013 Ampicillin SSSSSSSEEKKPPPPSEEPP pneumoniae A-9585 pyogenes A-960> aureus A-9537 aureus + 501 serum A-9537 aureus Pen-Res A-9606 aureus MethRes A15097 faecalis A20688 coli A15119 coli A20341 Pneumonia A15130 pneumoniae A20 + 68 mirabilis A-9900 vulgaris A21559 morganii A15153 rettgeri A21203 marcescens A20019 clcacae A-9659 cloacae A-9656 aeruginosa A-9843A aeruginosa A21213 125) 125 125> 125 125

>) 125

  ) 125> 125>) 125> 125> 125> 125> 125> 125> 125> 125> 125> 125 0,00+ 0,004 0,16 0,06> 125 0,13> 125) 125 0,13)> 125) 125) 125> 125

TABLE II

  Antibacterial activity of ceforamide and amoxicillin alone and in combination with BL-P2013 Beta- CIM compound (/ μg / ml) e Lactating organism ____

arnase C6fo- Céforanide Anoxi-

  ranide + BL-P2013 BL-P cillin Amoxi-

  (1: 1) 2013 + BL-P cilline (1: 1) B. fragilis

A21916

A22053

A22021

A21875

A22534

A22697

A22693

A22694

A22695

A22696

A22533

A22535

A22792

A22793

A22794

A22795

A22797

A22798

B. theto

taomicron

A22277

A22279

Bacteroides species

A20934

A21959

  + + + + + + + + + + + + + + + + + + + + + +) 125) 125> 125) 125> 125> 125 k 32 4- 4-> 125

2 8

3 .. 1+ 4- 4.> 125> 125> 125

63 2 16

63 4 16

32 2 8

32 2 16

  3. Antibacterial Activity of Ceforamide and Amoxicillin Isolenent and in Combination with BL-P2013 Good Synergy --- Marginal Synergy

  t The minimum inhibitory concentration (MIC) is determined

  was diluted on agar plate with diluted 2 + hEures cultures 50 times deposited with Steer's inoculator. The test medium consists of 5% Brucella agarose of lacquered sheep blood and 10 μg / ml of vitamin K. Beta-ICD (μg / ml)

amase Cefo- Céforanide Amoxi-

  ranide + BL-P2013 BL-P cillin Amoxi-

  (1: 1) 2013 + BL-P cilline (1: 1) Dacteroides species

A20929 + 63 _4 32 2 16

A21954 + 63 16 63 2 16

A20933 + 63 16 63 __ 8

A20930 + 125 8 125 4 32

A20931 + 63 4 32 2 16

A20927-1 - 0, 5 1 63 0.13 0.13

A20935 - 2 2,125 0.13 0.13

T A B L E A U III

  Therapeutic efficacy of amoxicillin in combination

  with the compound BL-P2013 in the experimentally infected mouse

  by a strain of staphylococcus aureus producing plactamase. Dosage: O and 2 hours drug administration

after the infection.

  DP50 infection / treatment (mg / kg) Organism (number (number.

of Organisms) Amoxicil-licine (A) BL-P2013 (B) A + B (1: 1)

IM PO IM PO IM PO

  S. aureus 5 x 108> 800> 800> 50> 200 6.3 44 A-9606 5 x 108> 800> 800> 50> 200 19 77

7 x 108> 800 -> 50 - 9.6 -

T AB L E A U IV

  Blood levels of the compound BL-P2013 and its pivaloyloxymethyl ester

  (BL-P2024) after oral administration to the mouse.

_ _, _

Blood level (/ ug / ml) Half

  Compound Dose 15 30 60 90 120 150 life Organism (mg / kg) Minutes after challenge (min)

4.5 5.1 3.9 2.3 1.5 0.8 50A 9675

A-9675

  BL-P2013 100 4.6 4.1 3 <2.6 <2.6 <2.6 - S. aureus

A-9606

  7, + 9.8 7.1 4.1 2.8 <2.6 50 S. aureus

A-9606

  12.8 12.9 9.7 7.2 5.2 4.2 70 E. coli

A-9675

  BL-P2024 100 13.1 12 8 5.7 3.8 <2.5 60 S. aureus

A-96060

  14.7 1.4 9.8 8.7 5.1 <2.5 60 S. aureus

A-9606

  These results are averages of two out of four trials. These results are averages of two out of four trials.

T A B L E A U V

  Blood level and half-life of BL-P2024 after administration by

  oral route in various doses to the mouse.

  BL-P2024 is suspended in a Tween-CMC-water mixture.

  The values are the average of 2 tests for doses of 50 mg / kg of 5 or 6 tests for the dose of 100 mg / kg and 2 or 3 tests for the dose of 200 mg / kg Test organism: E. coli A9675 for all doses except 200 mg / kg of BL-P2024

(S. aureus A9606).

  Blood level (/ ug / ml) Compound Dose 15 30 60 go90 120 150 life (mg / kg) vi (mg / kg> minutes after administration (min)

5.9 6.2 3.6 1.9 1.3 0.7 40

  BL-P2024 50 7.7 9.7 6.3 6.3 4.7 3.5 2.3 60

12.3 12.2 9.4 7.2 5.7 4.6 85

14.7 14.4 9.8 8.7 5.1 (2.5 60

ru% 0 o C>

T A B L E A VI

  Therapeutic efficacy of amoxicillin in combination with BL-P2013 in mice experimentally infected with S. aureus and E. coli strains producing β-lactamase Infection .... DP o / treatment (mg / kg) Infection _ _0 Organism (number of orAmoxicillin: BL-P2013 BLP systems) Amoxicillin 4: l 2: 1 1: 1 1: 2 l: 4 2013 S. aureus 7 x 108>) 200> 200:> 50 44:22 33:33 38:76 25:10) 200 A9606 2 x 109> 800 33:17 14:14 10:20> 200 S. aureus 4 x l08> 200 132: 66 78:39 4LF: 44> 200 A15091 5 x 108) 800 114: 57 100: 100 66: 132> 200 S. aureus 6 x 10O8) 200 174: 44+ 44+: 22 25:25 22:44 19:76 174+ A20379 5 x 108> 200 50:25 43:43 35:70) 200 E. coli 5 x 105> 200 25: 6.3 11: 5.5 11:11> 200 A20649 6 x 105) 200 20: 5 8: 4 6: 6 10:20 63:25) 200 E. coli 7 x 105> 200 6: 1.5 6: 3 5: 5) 200 A21223 6 x 105> 200 8: 2 6: 3 6: 6 4: 8 3.5: 1> 200 7 x 105> 200 7: 3.5 > 200 ru N oc :, -. TA 3 LEAU VI (cont'd) Therapeutic efficacy of amoxicillin in combination with co-exposure in patients with leukopenia in mice tested experimentally with 6 strains of S. aureus and. col producing 3-lactamrase infectionLDP0o / treatmern t mg / kg) Organism (number of orAmoxiciiline: BL-F2013 systems) BL-P ganisrnes) Amoxicillin4: 12: 1 1: 1 1: 2 1: 4 2013 E. coli 8 x 1o5> 200 8: 2 6: 3 3: 3 4: 8 +4: 16> 200 A9675 j 7 x 105> 100 10: 2, 56: 3 7: 7 2.5: 5> 100 6 x 105 > 400 13: 3.36: 3 5: 5 5:10> 200 Posology: Drugs are given orally O and 2 hours after infection

  The drugs are suspended in an aqueous Tween carboxymethylcellulose mixture.

ru -h. Nc

T A B L E A U VII

  Therapeutic efficacy of amoxicillin in various combinations with BL-P2013 pivaloyloxymethyl ester (BL-P2024) in mice experimentally infected with S. aureus and E. coli strains producing DP50-β-lactamase / treatment (mg / kg) a Infection 5 Organism (Excipient number of anatomy (AmoxiAmoxicillin excipient number: BL-P202) + BL-P of organisms) cillin 4: 1 2: 1 1: 1 2024 S. aureus 5 x 10 TCMC> 800 66:33 57:57> 200

A15091

  S. aureus 5 x 108 TCMC> 800 12: 6 9: 9,200

A20379

  E. coli 9 x 105 DMSO at> 100 12:12 6: 3 3: 3> 25

A9675 50%

  E. coli 7 x 105 DMSO at> 100 10: 2.5 7: 3.5 4: 4> 25

A9675 50%

  E. coli 7 x 105 TCMC> 100 8: 2 6: 3 5: 5 25 A9675 E. coli 6 x 105 TCMC> 400 9.2 7.2 2.4 7: 3.5 7: 7> 200 A9675 a Administration by Oral route O and 2 hours after infection

  b Amoxicillin: BL-P2024 + combinations are soluble in 50% dimethylsulfoxide.

  Amoxicillin is soluble in TCMC (Tween aqueous carboxymethylcellulose mixture) and

  BL-P2024 is administered in the suspension state in this exciplent.

cI co N -à

T A B L E A U VIII

  Oral Blood Levels in Mice of BL-P2036 and BL-P2013 or BL-P2024 Compounds nl Blood Levels (μg / ml) Compound Dose 15 30 60 90 120 150 (mg / kg) (mg / kg) Minutes after administration

  BL-P2026 25 1.7 2.3 0.9 <0.6 (0.6 0.6

(1.3-2.3) (2-2.6) (0.6-1.3)

BL-P2036 50 2 8 3.3 2.5 1.3 0.8

_

  (1.6 4.8) (2-5.3) (1.2-5.3) (0.7-2.4) (0.5-1.5) 0.6

BL-P2036 100 3 6 3.9 3.2 2 1.5 0.8

  (2.6 4.9) (2.9-5.1.) (2.4-4.4) (1.5-2.7) (1.1-2.1) (0.7- 1.0)

BL-P2013 25 1.3 1, 1.0 0

  (1.1-1.6) (1.2-1.6) (0.8-12) <0.6 <0.6 <0.6

  BL-P2013 50 2.2 2.6 1, 9 1, 5 1 1 0.8

  (1.8-2.7) (2.1-3.2 (1.4-2.6) (1.1-2.1) (0, -1.4) (0.5-1) , 3)

  BL-P2013 100 3.4 4.5 3.9 3.6 2.7 2.1

  (2.1-5.4) (3.6-5.7) (3.4-4.6) (2.5-5) (1.7-4.3) (1.4-3, 2)

  The compounds are presented in the Tween-CMC-H20 mixture. The values in parentheses are the 95% confidence limits. Test organism: E. coli A9675 (pH - 6.6, 0.1% inoculum) o -4 Test n 2 Blood level (/ ug / ml) Compound Dose 15 30 60 90 120 150 (mg / kg) Minutes after administration

  .-, = I..DTD: BL-P2024 25.307 43 2.3 0.8 0.6> 0.6

(2.6-5.5) (3-6.2), 0.9-5.8) O, 4-i, 8)

BL-P2024 50 5 4.5 4.9 3.6 2 6 17

  (0.8-33.3) (1.5-13.1 +) 2.4-9.8) 3.1-4.1) (1.5-4.3) (0.9-3 , 1)

_. _ ,,

  BL-P2024 100 8.3 9.5 7.8 4.8 3.9 3 7

  (5.5-12.6) (5.2-17.5) 4.2-14 +, 3) 3,4-6,7) (2,2-6,8) (2, A-5 )

BL-P2036 25 3.6 3.3 2.1 1.4 0.8

  (2.2-5.9) (2.2-4.7) 1.6-2.8) 0.7-2.8) (0, -1.7) <0.6

BL-P2036 50 4.0 3.6 2.2 2.1 1.5 1.0

  (3-5.3) (1,4-9) (1,4-3,7) 1,3-3,6) (0.6-3.6) (0.5-2, + 4)

,,, ._... J ,, ,,,,,

  BL-P2036 100 4.9 6.5 4, 5 2.8 1.9 1.3

  __ (3.7-6.5) (3.7-11.5) (3, 4-5.9) (2.2-3.6) (1-3.7) (0.4-4) )

  0) -) + The compounds are presented in 5% propylene glycol in the values in parentheses are the 95% confidence limits. Test organism: E. coli A9675 (pH = 6.6 inoculum 0.1%) ) mixture Tween-CMC-H20 ONO> - Test n 3 Blood levels (/ ug / ml) Compound Dose 15 30 60 90 120 50 (mg / kg) Minutes after administration

BL-P2013 25 1.4 2A0 1, 4 1.2

  | (1,1-1,9) (1,5-2,6) (0,9-2,1) (0,6-2,3) 1,2 (1,2

  BL-P2013 50 4, 6. 5, 5 3.6 3.1 2 1.1

  (3-6.9) (3-10.1) (2-6.4) (1.7-5.7) (0.8-5) (0.8-1.5)

  BL-P2013 100 6.8 11.5 6.4 4.2 3.3 2.5

  (14.8-9.6) 1 (7.3-1) (3.2-12.8) (2.1-8.6) (1.4-7; 6) (0.7-8) , 9)

  BL-P2036 25 4.3 4, 5 3.5 2, 5 2.2 2.0

  2.9-6.5) (2.2-9.2) (1.7-7.2) (1.3-4.8) (1-4.9) (1-3.7)

i

  BL-P2036 50 4.6 4.8 3.5 2, 2 1.9 1.8

  2,3-9,2) (3,3-7,1) (1,8-7) (1,2-4) (1,2-3) (0.3-4.6)

BL-P2036 100 7.1 6.7 5.8 4.3 2.7 2.7

  (3.1-16, 2) (3.6-12.3) (3.9-8.5) (3-6) (2.8-5, 6) (1.8-4)

  The compounds are presented in 5% propylene glycol in the Tween-CMC-HI mixture. The values in parentheses are the 95% confidence limits. Test organism: E. coli k9675 (pH = 6.6 inoculum of 0, The compounds of the invention are therefore useful, on oral or parenteral administration, to enhance the efficacy of O-lactam antibiotics against β-lactamase producing bacteria. . On a weight basis, the dose is one fifth to five times that of the 3-lactam antibiotics and is preferably equal to the latter. For example,

  compounds of the invention, as illustrated above, ad-

  In a 1: 1 ratio, the activity of ceforanide and amoxicillin was significantly improved against strains producing anaerobic Bacteroides flactamase, such as B. fragilis and B. thetaiotaomicron, in addition to other species of this genus, as well as against resistant forms of Staphylococcus aureus. The compounds of the invention are administered in admixture with or at the same time as the lactam antibiotic in a ratio dose.

  indicated with the known and usual dose of antibiotic

  tick. Therefore, the ability of the compounds of the invention to improve the effectiveness of a 3-lactam antibiotic against certain producing bacteria.

  P-lactamase makes them interesting for the administration.

  simultaneous treatment with certain 3-lactam antibiotics in the treatment of infections by bacteria in mammals and, in particular, humans. For the treatment of bacterial infection, a compound of the invention may be mixed with the 3-lactam antibiotic and both agents may be administered simultaneously. In

  Alternatively, a compound of the invention may be administered

  separately during treatment with a

P-lactam antibiotic.

  When a compound of the invention or a

  its salts is used to improve the antibacterial effect.

  it is not a β-lactam antibiotic, it can be administered as is or, preferably, with conventional pharmaceutical diluents or excipients. A compound of the invention in acid or pharmaceutically acceptable salt form can be administered orally or parenterally and a compound of the invention in ester form which is readily hydrolysed in vivo

  is most favorably administered orally.

  By parenteral administration, it is necessary to hear

  intramuscular, subcutaneous, intramuscular

peritoneal or intravenous.

  When a compound of the invention is used with a diluent or excipient, it is chosen according to the intended mode of administration. For example, for oral administration, the compound may be presented in the form of tablets, capsules, lozenges, lozenges, powders, syrups, elixirs, aqueous solutions or suspensies, etc. . following current pharmaceutical practice. The relationship between

  the active constituents and the excipient obviously depend on

  the chemical nature, solubility, stability and effectiveness of the active ingredients,

  as well as the dose envisaged. Nevertheless, these compounds

  pharmaceutical compositions are likely to contain about 5 to 80% of excipient. In the case of tablets

  for oral use, commonly used excipients are

  lactose, sodium citrate and phos-

  phosphates. Various agents of disintegration, such as

  donation and lubricating agents, such as magnesium stearate

  sodium lauryl sulphate and talc are used.

  frequently used in tablets. For the administration

  oral administration in the form of capsules, di-

Useful ingredients are lactose and polyethylene glycol

  high molecular weight passes. In aqueous suspensions administered orally, the active ingredients are combined with emulsifiers and

  suspending. If the thing is desired,

  some sweetening and / or flavoring agents may

  to be added. For the parent administration

  particularly intramuscular, intraperitoneal, subcutaneous or intravenous

  usually presented in the form of sterile solutions

  riles whose pH is advantageously adjusted by means of a buffer. For intravenous administration, the

  total concentration of solutes should make the

isotonic position.

  Although the physician ultimately decides on the dose administered to his patient, the ratio of the daily dose of a compound of the invention or its salt to the daily dose of a beta-lactam antibiotic is normally about 1: 5 to 5: 1 and, of

  preferably about 1: 1. In addition, the daily dose

  of each active ingredient is normally

  viron 10 to 200 mg per kg of body weight in administration

  oral administration and approximately 10 to 100 mg per kg

  of body weight in parent administration

  rale. These values are however only indicative and it may sometimes be necessary to administer doses

falling outside these limits.

  The invention is susceptible of industrial application.

Claims (11)

  1- Ester of formula: CH3 R-CH-CO-Nil NH2 It C H2C1   ## STR2 ## wherein R 1 represents a hydrogen atom or a hydroxyl radical and R represents a hydrogen or chlorine atom or a   hydroxyl, methyl or methoxy radical.   2. Esters according to claim 1 of the acid of formula: ## STR2 ##   characterized in that the alcoholic part is   presented by 6 - [(R) -2-amino-2-phenyl-   acetamido-3,3-dimethyl-7-oxo-4-thia-l-aza-bicyclo [3.2.03   heptane-2-carbonyl-oxynethyl or 6 - [(R) -2-amino-2-p-   hydroxyphenylacetamido] -3,3-dimethyl-7-oxo-4-thia-1-aza-bicyclo [3.2.0] heptane-2-carbonyloxymethyl. 3. Process for preparing the following compounds   claims 1 or 2, characterized in that at stages   successive, a) is subjected to catalytic hydrogenation an ester of formula: CHCl DD 2   CH3 CO2R o R represents a benzyl or substituted benzyl radical; b) the hydrogenated product is subjected to oxidation to form the desired acid or its salt, then, if desired, c) the acid or salt is esterified to produce   an easily hydrolysed ester of the acid.   4. Process according to claim 3, characterized   in that the hydrogenation is carried out with the aid of a palladium catalyst.   5. Process according to claim 3, characterized   in that the oxidation is carried out by means of a   alkali metal permanganate or an organic peracid.   6. Process for preparing the following compounds   claims 1 or 2, characterized in that at stages   successive, a) is oxidized, for example by means of KMnO4, H202 or a similar peroxide or a peracid, an ester of formula: 2491C71 CHCl -3 C-OCHLC   It forms an ester sulfoxide of the formula: ## STR2 ## then, b) this ester sulfoxide is reacted with a metal in an acid, such as zinc in glacial acetic acid, to produce the desired acid. or salt, then, if desired, c) the acid or its salt is esterified to produce   an easily hydrolysed ester of the acid.   7. Process for the preparation of the compounds according to claim 1, characterized in that an acid is reacted with a solution of a compound of formula: CH3 R- CH CO-NH NOT R- C H Il Ri-c / N o0 C = O 1 = 0 CH2C1 1 CH3 CH2 C ... 0   R represents: R5 9'X'emeU.4 o R represents a hydrogen atom or a hydroxyl radical and R5 represents a hydrogen or chlorine atom or a hydroxyl, methyl or methoxy radical; R1 represents an alkyl, aralkyl or aryl radical; R2 represents a hydrogen atom or an alkyl, aralkyl or aryl radical and R3 represents an alkyl, aralkyl, aryl or alkoxy radical, 4 5 4 5   aralkoxy or aryloxy or -NR4R5, where R and R each represent a hydrogen atom or an alkyl, aralkyl or   aryl, or taken together with the nitrogen atom, represent   a piperidino or morpholino radical, which reaction with the acid is carried out in an organic solvent or in aqueous or partially aqueous solution at room temperature. 8. Process according to Claim 7, characterized in that R1 represents a methyl radical, R2 represents a hydrogen atom and R3 represents a radical. methoxy, ethoxy or methyl.   9. Process according to Claim 7, characterized in that the reaction is carried out in acetone or chloroform.   10. Process according to claims 7 to 9,   characterized in that the reaction is carried out at a pH of 1 to 5.   11. Antibacterial composition, characterized in that it comprises as active ingredient, a following ester one of claims 1 or 2.