DK155740B - METHOD OF ANALOGUE FOR PREPARATION OF PENICILLANIC ACID-1,1-DIOXIDE, CERTAIN ESTERS AND PHYSIOLOGICALLY ACCEPTABLE SALTS - Google Patents

Description

•. DK 155740 B

in

The present invention relates to an analogous process for the preparation of novel penicillanic acid derivatives of formula I

O5H3 '*

J-N-L

Q <% COORb 6 wherein R is hydrogen, 3-phthalidyl, 4-crotonolactonyl, γ-butyro-lacton-4-yl or a group of formula f i? 5 f * S,

-ά-O-C-R5 X or -C-0-C-0-R5 XI

wherein R and R are each hydrogen, methyl or ethyl and R is alkyl

C

with 1-6 carbon atoms, and when R represents hydrogen, physiologically acceptable salts thereof, and the process is characterized by the characterizing part of claim 1.

Various convenient embodiments of the method according to the invention may be used as set forth in claims 2-7.

One of the most well-known and commonly used classes of antibacterial agents is the so-called β-lactam antibiotics. These compounds are characterized in that they have a core consisting of a 2-aetidinone ring (β-lactam ring) condensed with either a thiazole dinar ring or a dihydro-1,3-thiazine ring. When the core contains a thiazolidine ring, the compounds are usually referred to by the common name penicillins, while the compounds in which the core contains a dihydrothiazine ring are referred to as cephalosporins. Typical examples of penicillins commonly used in clinical practice are benzylpenicillin (penicillin G), phenoxymethylpenicillin (penicillin V), ampicillin and carbenicillin, and typical examples of common cephalosporins are cephalothin, cephalexin and cefazoline.

However, despite the widespread use and widespread recognition of the β-lactam antibiotics as valuable chemotherapeutic agents, they have the significant disadvantage that some members are not active against certain microorganisms. It is believed that this resistance of a particular microorganism to a given β-lactam antibiotic is in many cases due to the microorganism producing 2

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and β-lactamase. The latter substances are enzymes that cleave the 3-lactam ring into penicillins and cephalosporins to form products devoid of antibacterial action. However, certain substances have the ability to inhibit β-lactamases, and when a β-lactamase inhibitor is used in combination with a penicillin or cephalosporin, it may increase or enhance the antibacterial efficacy of the penicillin or cephalosporin against certain microorganisms. An increase in antibacterial efficacy is expected when the antibacterial effect of a combination of a 3-lactamase inhibitor and a 3-lactam antibiotic is significantly greater than the sum of the antibacterial effects of the individual components.

The process of the invention provides certain novel chemical compounds of Formula I which are novel members of the class of antibiotics referred to as the penicillins and which are useful as antibacterial agents. More specifically, the subject pencillin compounds are penicillanic acid 1,1-dioxide and in vivo readily hydrolyzable esters thereof.

In addition, penicillanic acid 1,1-dioxide and its in vivo readily hydrolyzable esters are potent inhibitors of microbial β-lactamases.

1,1-Dioxides of benzylpenicillin, phenoxymethylpenicillin and certain esters thereof are disclosed in U.S. Patent Nos. 3,197,466 and 3,536,698 and in an article by Guddal et al., In Tetrahedron Letters, 9, 381 (1962). . Harrison et al., In the Journal of the Chemical

Society (London), Perkin I, 1772 (1976), has described several different penicillin-1,1-dioxides and -1-oxides, including methyl-phthalimidopenicillinate-1,1-dioxide, methyl-6,6-dibrompenicillanate -1,1-dioxide, methyl-penicillanate-1-oxide, methyl-penicillanate-13-oxide, 6,6-dibrompenicillanic acid-1-oxide and 6,6-dibrompenicillanic acid-13-oxide.

The process of the invention provides novel compounds of the above Formula I wherein

C

R has the above meaning. The aforementioned falling groups under the definition g for R are readily hydrolyzable ester forming groups in vivo. These "in vivo readily hydrolyzable ester forming groups" are nontoxic ester groups that are readily cleaved in mammalian blood or tissue to release the corresponding free acid (i.e.

C.

the compound of formula I wherein R is hydrogen).

The compounds of the invention

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3 binders of formula I are useful as antibacterial agents and for enhancing the antibacterial effect of β-lactam antibiotics.

The starting compounds used in process a) according to the invention have formulas II, III and III1 2

ISLAND ISLAND

1 \ ifs I — Y f-c "i ... I-Y f-" ".

J-N- \! ^ - 'N- \ 1

CT 'COOR Q> COOR

H "CH-% S / 3 or - III III y - N - k, in or" coor 1 6 in which formulas R has the same meaning as R or is a conventional penicillin carboxy protecting group. The compounds of formulas II and III are otherwise novel compounds.

The present invention relates to a process for preparing the novel compounds of formula I, and throughout the present disclosure these compounds are referred to as penicillanic acid derivatives which may be reproduced by structural formula r4 "V3

PCH 3 IV

-N-W.

Formula IV indicates the association of a substituent to the bicyclic nucleus with a broken line that the substituent is below the plane of the bicyclic 2 nucleus. Such a substituent is said to be in α-configuration. Conversely, the association of a substituent to the bicyclic 3 sphere core with a fully drawn line indicates that the substituent is located above the plane of the core. This last configuration is referred to as (3 configuration).

4 DK 155740 B! The present disclosure also discloses certain derivatives of cephalosporanoic acid having the formula

H

rjf ^ i.

QT N | ^ iss ^ CH2-0-C-CH3 V

COOH

In formula V, the hydrogen atom at C-6 is below the plane of the bicyclic nucleus. The derived terms desacetoxycephalosporanoic acid and 3-desacetoxymethylcephalosporanoic acid are used for compounds of structures VI and VII, respectively.

i-h i-0 ^ -N \ j ^ CH3 0 ^ -

COOH COOH

VI VII

4-Crotonolactonyl and Y-butyrolacton-4-yl refer to Structures VIII and IX, respectively. The wavy lines must denote each of the two epithets and mixtures thereof.

o 0

VIII IX

c

The above groups falling under R are well known in the art of penicillin, see e.g. German Publication Publication No. 2,517,316. In many cases, they improve the absorption properties of the penicillin compound. Preferred groups for R are alkanoyloxy methyl having from 3 to 8 carbon atoms, 1- (alkanoyloxy) ethyl having from 4 to 9 carbon atoms, 1-methyl-1- (alkanoyloxy) ethyl having from 5 to 10 carbon atoms, 3-phthalidyl, 4 -crotonolactonyl and γ-butyro-lacton-4-yl.

"DK 155740B

5 g

The compounds of formula I wherein R has the above meaning can be prepared by process a) of the invention by oxidation of each of the compounds of formula II or III, wherein R has the above meaning. Many different oxidizing agents known in the art for the oxidation of sulfoxides to sulfones can be used for this process.

However, particularly convenient reagents are metal permanganates such as alkali metal permanganates and alkaline earth metal permanganates, and organic peroxy acids such as organic peroxycarboxylic acids. Suitable simple wood reagents are sodium permanganate, potassium permanganate, 3-chloroperbenzoic acid and peracetic acid.

When a compound of formula II or III, wherein R is as defined above, is oxidized to the corresponding compound of formula I using a metal permanganate, the reaction is usually carried out by treating the compound of formula II or III with from ca. 0.5 to approx. 5 molar equivalents of the permanganate, and preferably approx. 1 molar equivalent of the permanganate, in a suitable solvent system. A suitable solvent system is one which does not adversely react with either the starting materials or the product and water is generally used. If desired, a co-solvent which is miscible with water but which does not react with the permanganate such as tetrahydrofuran can be added. The reaction is usually carried out at a temperature in the range of approx. -20 to approx. 50 ° C, and preferably at ca. 0 ° C. At about. 0 ° C, the reaction is usually substantially complete within a short period of time, e.g. Within 1 Hour. Although the reaction may be carried out under neutral, basic or acidic conditions, it is preferred to work under substantially neutral conditions to avoid decomposition of the β-lactam ring system in the compound of formula I. Indeed, it is often advantageous to buffer the reaction medium to a pH. value near the neutral point. The product is extracted in a conventional manner. Any excess of manganese is usually decomposed using sodium hydrogen sulfite and then the product, if not in solution, is recovered by filtration. It is separated from manganese dioxide by extraction of it in an organic solvent and removal of the solvent by evaporation. Alternatively, the product, if present in solution, is isolated by the usual solvent extraction process.

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When a compound of formula II or III wherein R is as defined above is oxidized to the corresponding compound of formula I using an organic peroxy acid, e.g. a peroxy-carboxylic acid, the reaction is usually carried out by treating the compound of formula II or III with from ca. 1 to approx. 4 molar equivalents, and preferably approx. 1.2 equivalents of the oxidant in a reaction-inert organic solvent. Typical solvents are chlorinated hydrocarbons such as dichloromethane, chloroform and 1,2-dichloroethane, and ethers such as diethyl ether, tetrahydrofuran and 1,2-dimethoxyethane. The reaction is usually carried out at a temperature of from ca. -20 to approx. 50 ° C, and preferably at ca.

25 ° C. At about. Reaction times of about 25 ° C are generally used.

2 to approx. 16 hours. The product is usually isolated by removing the solvent by evaporation in vacuo. The product can be purified by conventional means.

When oxidizing a compound of formula II or III to a compound of formula I using an organic peroxy acid, it is sometimes advantageous to add a catalyst such as a manganese salt, e.g. manganiacetylacetonat.

When, by the oxidation of process a) of the invention, a compound of formula I is obtained with the exception that g is substituted for a pencillin carboxy protecting group 1 in place of g, this protecting group R is removed to give the compound. of formula I wherein R is hydrogen. In this context, 1R may be any carboxy protecting group conventionally used in the penicillin range for protecting carboxy groups at the 3-position. The identity of the carboxy protecting group is not essential. The only requirements for the carboxy protecting group R 1 are that: (i) it must be stable during the oxidation of the compound of formula II or III, and (ii) it must be removable from the compound of formula I using conditions under which β The lactam remains essentially intact. Typical examples of groups that can be used are the tetrahydropyranyl group, the benzyl group, substituted benzyl groups (eg 4-nitrobenzyl), the benzhydryl group, 2,2,2-trichloroethyl group, the t-butyl group and the phenazyl group. See also U.S. Patent Nos. 3,632,850 and 3,197,466, British Patent No. 1,041,985, Woodward et al., Journal of the American Chemical Society, (38, 852 (1966), Chauvette, Journal of Organic Chemistry, 36, 1259 (1971), Sheehan et al., Journal of

7 DK 155740 B

Organic Chemistry, 29, 2006 (1964), and "Cepahalosporin and Penicillins Chemistry and Biology", edited by H.E.Flynn, Academic Press, Inc./ 1972. The penicillin carboxy protecting group is removed in a conventional manner, taking due account of the lactamation of the 3-lactam system.

Similarly, compounds of formula g I can be prepared in which R has the above meaning by oxidizing a compound of formula III 'wherein R has the above meaning. This oxidation is carried out in exactly the same manner as described above for the oxidation of a compound of formula II or III, except that twice as much oxidant is usually used.

Process b) according to the invention consists in the preparation of compounds of formula I wherein R has the above meaning other than hydrogen, reacting a salt of pencillanic acid 1.1-dioxide with 3-phthalidyl chloride, 3-phthalidyl bromide, 4-crotono-lacton-4-yl chloride, 4-crotonolacton-4-yl bromide, γ-butyrolacton-4-yl bromide or a compound of the formula

? ? R3 O

I II s I II 5

Q-C-G-C-R XII or Q-C-0-C-0-R3 XIII

wherein R -R has the above meanings, and Q is chlorine or bromine, in a reaction-inert solvent at a temperature in the range of 0 to 100 ° C.

The reaction is conveniently carried out by dissolving a salt of penicillanic acid 1.1 dioxide in a suitable polar organic solvent such as Ν, Ν-dimethylformamide, and then adding approx. one mole equivalent of that chloride or bromide. When the reaction is substantially complete, the product is isolated in the usual manner. It is often sufficient simply to dilute the reaction medium with an excess of water and then extract the product into a water-immiscible organic solvent and then recover it by evaporation of the solvent. The commonly used salts of penicillanic acid 1,1-dioxide are alkali metal salts such as sodium and potassium salts, and tertiary amine salts such as triethylamine, N-ethylpiperidine, Ν, Ν-dimethylaniline and N-methylmorpholine salts. The reaction is carried out as mentioned at a temperature in the range of 0 to 100 ° C, 8

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and usually approx. 25 ° C. The time required to complete the reaction varies with several different factors, such as the concentration of the reactants and the reactivity of the reagents. When considering the halogen compound, the bromide reacts faster than the chloride. When using a chlorine compound, it is sometimes advantageous to add up to one mole equivalent of an alkali metal iodide. This causes an increase in the reaction rate. Taking full account of the above factors, reaction times of approx. 1 to approx. 24 hours.

Penicillanic acid la-oxide, the compound of formula II wherein R 2 is hydrogen, can be prepared by debromating 6,6-dibrompenic-cillanic acid la-oxide. The debromation can be carried out using a conventional hydrogenolysis method. Thus, a solution of 6,6-dibrompenicillanic acid oxide is stirred or shaken under an atmosphere of hydrogen, or hydrogen mixed with an inert diluent such as nitrogen or argon, in the presence of a catalytic amount of palladium-on-calcium carbonate catalyst. . Suitable solvents for this debromation 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 solvents. However, conditions are usually chosen under which the dibromo compound is soluble. The hydrogen genolysis is usually carried out at room temperature and at a pressure of approx. atmospheric pressure to approx. 3.5 at. The catalyst is usually present in an amount of approx. 10% by weight, based on the dibromo compound, and up to an equal amount of weight as the dibromo compound, however larger quantities can be used. The reaction usually takes approx. 1 hour, after which the compound of formula II wherein R 1 is hydrogen is simply recovered by filtration followed by removal of the solvent in vacuo.

6,6-Dibrompenicillanic acid la-oxide is prepared by oxidation of 6,6-dibrompenicillanic acid with an equivalent of 3-chloroperbenzoic acid in tetrahydrofuran at 0 to 25 ° C for approx. 1 hour following the method set forth by Harrison et al., Journal of the Chemical Society (London) Perkin I, 1772 (1976). 6,6-Dibrompenicillanic acid is prepared by the method of Clayton,

Journal of the Chemical Society (London), (C) 2123 (1969).

Penicillanic acid Iβ3 oxide, the compound of formula III wherein in R is hydrogen, can be prepared by controlled oxidation of γ

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pennicillanic. Thus, it can be prepared by treating penicillanic acid with one mole equivalent of 3-chloroperbenzoic acid in an inert solvent at ca. 0 ° C for approx. 1 hour. Typical solvents which may be used include chlorinated hydrocarbons such as chloroform and dichloromethane, ethers such as diethyl ether and tetrahydrofuran, and low molecular weight esters such as ethyl acetate and butyl acetate. The product is extracted in a conventional manner.

Penicillanic acid is prepared as described in British Patent Specification No. 1.07 2,108.

. in

Compounds of formulas II and III in which R has the same meaning as R except hydrogen can be prepared directly from the compound of formula II or III wherein R is hydrogen by esterification, i. by alkylating said compound with the formula II or III wherein R is hydrogen, with a 3-phthalidyl halide, a 4-crotonolactonyl halide, a Y-butyrolacton-4-yl halo or a compound of formula XII or XIII. The reaction is carried out in exactly the same manner as described above for process b) of the invention.

in

Alternatively, the compounds of formula II, wherein R has g the same meaning as R other than hydrogen, can be prepared by oxidation of the corresponding ester of 6,6-dibrompenicillanic acid followed by debromation. The esters of 6,6-dibrompenicillanic acid are prepared from 6,6-dibrompenicillanic acid by conventional methods. The oxidation is carried out e.g. by oxidation with one molar equivalent of 3-chloro-perbenzoic acid as described above regarding the oxidation of 6,6-dibrompenicillanic acid to 6,6-dibrompenicillanic acid 1a-oxide and the debromation is carried out as described above regarding the debromation of 6,6-dibrompenicillanic acid-1a oxide.

Similarly, the compounds of formula III wherein R 6 has the same meaning as R except hydrogen can be prepared by oxidation of the corresponding ester of penicillanic acid. The latter compounds are readily prepared by esterification of penicillanic acid using conventional methods. The oxidation is carried out e.g. by oxidation with one mole equivalent of 3-chloroperbenzoic acid as described above regarding the oxidation of penicillanic acid to penicillanic acid 16 oxide.

The compounds of formula II wherein R 1 is a carboxy protecting group can be obtained in two ways. They can be obtained simply by

, 0 DK 155740 B

take penicillanic acid-la-oxide and attach a carboxy protecting group thereto. Alternatively, they may be obtained by: (a) associating a carboxy protecting group with 6,6-dibrompenicillanic acid, (b) oxidizing the protected 6,6-dibrompenicillanic acid to a protected 6,6-dibromo-penicillanic acid la-oxide using one mole equivalent of 3-chloro-perbenzoic acid, and (c) debromate the protected 6,6-dibrompenicillanic acid la-oxide by hydrogenolysis.

The compounds of formula III wherein R 1 is a carboxy protecting group can be obtained simply by attaching a protecting group to penicillanic acid ip oxide. Alternatively, they may be obtained by: (a) associating a carboxy protecting group with penicillanic acid, and (b) oxidizing the protected penicillanic acid using one molar equivalent of 3-chloroperbenzoic acid as described above.

6 1

The compounds of formulas I, II and III in which R R is hydrogen, is acidic and forms salts with basic substances. Such salts are included in the scope of the invention. These salts can be prepared by conventional methods, such as contacting the acidic and basic components, usually in a molar ratio of 1: 1, in an aqueous, non-aqueous or partially aqueous medium as the case may be. They are then recovered by filtration, by precipitation with a non-solvent followed by filtration, by evaporation of the solvent or, in the case of aqueous solutions, by lyophilization, as the case may be. Basic substances which are suitably used for salt formation belong to both the organic and inorganic type and include ammonia, organic amines, alkali metal hydroxides, carbonates, hydrogen carbonates, hydrides and alkoxides, and alkaline earth metal hydroxides, carbonates, hydrides and alkoxides. Representative examples of such bases are primary amines such as n-propylamine, n-butylamine, aniline, cyclohexylamine, benzylamine and octylamine, secondary amines such as diethylamine, morpholine, pyrrolidine and piperidine, tertiary amines such as triethylamine, N-ethylpiperidine, N-methylmorpholine and 1,5-diazabicyclo [4,3,0] non-5ene, hydroxides such as sodium hydroxide, potassium hydroxide, ammonium hydroxide and barium hydroxide, alkoxides such as sodium ethoxide and potassium ethoxide, hydrides such as calcium hydride and sodium hydride, carbonates such as potassium carbonate and sodium carbonate, hydrogen carbonates such as sodium hydrogen carbonate and potassium hydrogen carbonate, and alkali metal salts of long chain fatty acids such as sodium 2-ethyl hexanoate.

,, DK 155740 B

Preferred salts of the compounds of formulas I, II and 6 1 III wherein R R is hydrogen, is sodium, potassium and triethylamine salts.

As indicated above, the compounds of the formula I of the compound of the invention are of medium strength. The in vitro action of the compound of formula I wherein R is hydrogen can be detected by measuring its minimum inhibitory concentrations (MIC) in µg / ml against several different microorganisms. The procedure followed is that recommended by the International Collaborative Study on Antibiotic Sensitivity Testing (Ericcson and Sherris, Acta Pathologica et Microbiologia Scandinav., Supp. 217, sections A and B: 1-90 [1970]), and by this is used for the brain-heart infusion agar (BHI agar) and the graft repeat device. The tube glass, which has been growing overnight, is diluted 100 times for use as a standard graft (20,000 to 10,000 cells in approximately 0.002 ml are applied to the agar surface; 20 ml of BHI agar / dish). Twelve dilutions of the test compound are used twice, with the initial concentration of the test drug being 200 µg / ml. Single colonies are ignored when the plates are read after 18 hours at 37 ° C. The sensitivity of the test organism (MIC) is taken as the lowest concentration of the compound capable of producing complete growth inhibition as judged by the unarmed eye. MIC values for penicillanic acid.e-1,1-dioxide against a variety of microorganisms are listed in Table I.

12 DK 155740B '

Table I

In vitro antibacterial action of penic ± llanoic acid -1,1-dioxide.

Microorganism '.....' 'MIC (y'g / ml _

Staphylococcus aureus 100

Streptococcus faecalis> 200

Streptococcus pyogenes 100

Escherichia coli 50

Pseudomonas aeruginosa 200

Klebsiella pneumoniae 50

Proteus mirabilis 100

Proteus morgani 100

Salmonella typhimurium 50

Pasteurella multocida 50

Serratia marcescens 100

Enterobacter aerogenes 25

Enterobacter clocae 100

Citrobacter freundii 50

Providencia 100

Staphylococcus epidermis 200

Pseudomonas putida> 200

Hemophilus influenzae> 50

Neisseria gonorrhoeae '...... 0.312 ......

The compounds of formula I prepared by the process of the invention act as antibacterial agents in vivo. To determine this effect, acute experimental infections in mice are induced by intraperitoneal inoculation on the mouse of a standard culture of the test organism suspended in 5% porcine mycin. The severity of the infection is standardized so that the mice receive one to ten times the LD 1Q dose of the organism (LDiqq: the minimal amount of inoculation with the organism required to consistently kill 100% of the infected, untreated control mice). The test compound is administered to the infected mice using a multiple dosing system. At the end of the test, the effect of a compound is assessed by counting the number of survivors among the treated animals and expressing the effect of a compound as the percentage of animals that survived.

The in vitro antibacterial effect of the compound of formula I wherein R is hydrogen makes it useful for local use as a disinfectant. In the case of using this compound for topical use, it is often convenient to mix the active ingredient with a non-toxic carrier such as vegetable or mineral oil or a softening cream. Likewise, it can be dissolved or dispersed in liquid diluents or solvents such as water, alkanols, glycols or mixtures thereof. In most cases, it is appropriate to use concentrations of the active ingredient of from ca. 0.1 to about 10% by weight, based on the total composition.

The in vivo effect of the compounds of formula I prepared by the process of the invention makes it suitable for controlling bacterial infections in mammals, and including humans, by administration by both oral and parenteral routes. The compounds can be used to control infections caused by sensitive bacteria in humans, e.g. infections caused by strains of Neisseria gonorrhoeae.

When considering the therapeutic use of a compound of formula I, or a salt thereof, in a mammal, especially a human, the compound may be administered alone or it may be admixed with pharmaceutically acceptable carriers or diluents.

They can be administered orally or parenterally, ie. intramuscularly, subcutaneously, or intraperitoneally. The carrier or diluent is selected based on the intended mode of administration. For example, when In view of the oral mode of administration, an antibacterial penam compound prepared by the method of the invention can be used in the form of tablets, capsules, lozenges, dulciblettae, powders, syrups, elixirs, aqueous solutions and suspensions and the like in accordance with conventional pharmaceutical practice. The amount ratio of active ingredient to carrier will, of course, depend on the chemical nature, solubility and stability of the active ingredient, as well as the intended dosage. However, pharmaceutical compositions containing an antibacterial compound of formula I are likely to contain from ca. 20 to approx. 95% active ingredient.

In the case of tablets for oral use, the carriers commonly used include lactose, sodium citrate and salts of phosphoric acid. Various disintegrants such as starch and lubricants such as magnesium stearate, sodium lauryl sulfate and talc are commonly used in tablets. For oral administration

, 4 DK 155740 B

capsule form are useful diluents lactose and high molecular weight polyethylene glycols. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweeteners and / or flavors may be added. For parenteral administration, including intramuscular, intraperitoneal, subcutaneous and intravenous use, sterile solutions of the active ingredient are usually prepared and the pH of the solutions is adjusted and appropriately buffered. For intravenous use, the total concentration of solutes must be adjusted so that the preparation is isotonic.

As previously stated, antibacterial agents containing the compounds of the present invention can be used in humans against sensitive organisms. The prescribing physician will ultimately determine the right dose for a particular person, and this can be expected to vary according to the individual patient's age, weight and response, as well as the nature and severity of the patient's symptoms. The present compounds will normally be used orally in doses ranging from about 10 to approx. 200 mg per kg body weight per daily and parenterally in doses from ca. 10 to approx. 400 mg per day kg body weight per day. However, these figures are for illustrative purposes only and in some cases dosages outside these limits may be necessary.

However, as indicated above, the compounds of formula I prepared by the process of the invention are potent inhibitors of microbial β-lactamases and increase the antibacterial efficacy of β-lactam antibiotics (penicillins and cephalosporins) against many microorganisms, especially those producing a β-lactamase. lactamase. The manner in which said compounds of formula I increase the efficacy of a 3-lactam antibiotic can be assessed by experiments measuring MIC for a given antibiotic alone and for a compound of formula I alone.

These MICs are then compared to the MICs obtained with a combination of the given antibiotic and the compound of formula I. When the antibacterial potency of the combination is significantly greater than could be predicted from the potency values of the individual compounds, it is considered to constitute an effect increase. The MIC values for combinations are measured using the method described by Barry and Sabath in the "Manual of Clini-

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cal Microbiology ", published by Lenette, Spaulding and Truant, 2nd ed., 1974, American Society for Microbiology.

Experimental results showing that penicillanic acid 1,1-dioxide increases the efficacy of ampicillin (6- (D-2-amino-2-phenylacetamido) penicillanic acid) are listed in Table II. From Table II, it can be seen that against 19 ampicillin-resistant strains of Staphylococcus aureus, the type value of MIC for ampicillin and for penicillanic acid 1,1-dioxide is 200 µg / ml. However, the MIC type values for ampicillin and penicillanic acid-1,1-dioxide in combination are 1.56 and 3.12 µg / ml, respectively. Put another way, this means that while ampicillin alone has a MIC value of 200 µg / ml against 19 strains of Staphylococcus aureus, its type value of MIC is reduced to 1.56 µg / ml in the presence of 3.12 µg / ml periicillansyre-1,1-dioxide. The other entries in Table II show the increase in the antibacterial efficacy of ampicillin against 26 ampicillin-resistant strains of Haemophilus influenzae, 18 ampicillin-resistant strains of Klebsiella pneumoniae and 15 strains of the anaerobic Bacteroides fragilis. Tables III, IV and V show the increase in the antibacterial potency of benzylpenicillin (penicillin G), carbenicillin (di-carboxybenzylpenicillin) and cefazoline (7- (2- [1-tetrazolyl) acetamido-3- (2- [5- methyl-1,3,4-thiadiazolyl] -thiomethyl-3-desacetoxymethylcephalosporanoic acid) against strains of S. aureus, H. influenzae, K. pneumoniae and Bacteroides fragilis.

16

DK 155740 B

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18 DK 1 55740 B

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DK 155740B

Additional biological test data for five representative compounds A to F as defined below are set forth in Experiments A to E below, where Experiments A, B and E demonstrate that compounds B, C, D and F are hydrolyzed to penicillanic acid 1,1-dioxide after administration to mice, while Experiments C and D demonstrate that compounds B and E are effective in increasing the antibacterial efficacy of ampicillin.

STRUCTURES

H CH

J LJ 3

</ /// COOR

Compound A: R is H

Compound B: R is CH 2 -O-CO-C (CH 3) 3

Compound C: R is 100

Compound D: R is CH (CH 3) -O-CO-O-CH 2 CH 3

Compound E: R is C (CH3) 2-O-CO-CH3

Compound F: R is CH 2 -O-CO-0-CH 3

RESULTS

Experiment A

Compounds A (as its sodium salt), B and C were orally administered to mice at a single dose of 20 mg / kg. Blood samples were taken on the mice at intervals starting at 15 minutes after dosing. The blood samples were bio-tested for anti-bacterial action using the microorganism Comamonas terrigena. The results are listed in Table VI. In Table VI, the antibacterial effect in the blood samples is expressed as the concentration of compound A in µg / ml.

DK 155740B

21

TABLE VI

Sampling time Antibacterial action in blood (min) (as pg / ml of compound A)

Compound A Compound B Compound C

1.52 + 0.62 2.87 + 0.83 3.16 + 1.03 2.48 + 0.64 3.06 + 0.24 2.48 + 1.28 60 1.80 + 0 , 46 1.78 + 0.33 0.90 + 0.90 .90 0.87 + 0.53 0 0.59 + 0.49 a * 120 00 0

Experiment B

Compounds A and D were administered orally to mice at a single dose of 20 mg / kg and 50 mg / kg. Blood samples were taken on the mice at intervals starting at 15 minutes after dosing. The blood samples were bio-tested for antibacterial activity using the microorganism Comamonas terrigena. The results are listed in Table VII. In Table VII, the antibacterial effect in the blood samples is expressed as the concentration of compound A in vg / ml.

TABLE VII

Sampling Antibacterial activity in blood time (as µg / ml of compound A) (min) -----—-- _ 20 mg / kg

Compound A Compound D Compound A Compound D

1.97 + 0.88 4.92 + 0.38 6.03 + 1.24 10.9 + 0.87; 2.40 + 1.00 4.16 + 0.22 7.02 + 1.26 7.90 + 1.00 45 2.54 + 1.06 2.56 + 0.66 6.90 + 1, 38 5.68 + 0.95 60 2.37 + 0.97 0 5.72 + 1.67 4.58 + 0.68 75 2.03 + 0.91 0 5.66 + 2.14 3.77 + 0.47 90 1.84 + 0.75 0 3.20 + 1.76 2.68 + 0.76

22 DK 155740B

Experiment C

The in vivo antibacterial effect of ampicillin, compound B and a 1: 1 mixture of ampicillin and compound B was measured against three strains of ampicillin-resistant Staphylococcus aureus in mice. Acute experimental infections were induced in mice by intraperitoneal inoculation on the mice by a standard culture of Staphylococcus aureus suspended in 5% pig stomach mucin. The severity of the infection was standardized such that the mice received one to ten times the LD 1Q dose of the organism (LD10Q: the minimal amount of graft with the organism required to consistently kill 100% of the infected, untreated control mice). The test compound was administered subcutaneously (sc) or orally (po) by a multiple dosing system at which the first dose was given 0.5 hours after inoculation and repeated 4 and 24 hours later. The number of surviving mice was then determined 96 hours after inoculation. The results are listed in Tables VIII, IX and X. The effects are given as PDj-g values (PD ^: the amount of drug that protects 50% of the mice).

TABLE · VIII

PD50 g) mod

Compound Staphylococcus aureus 01A182 PO sc ampicillin> 100> 100

Compound B> 100> 100 ampicillin + (8.45 ± 1.9) + (8.14 + 0.9) +

Compound B (8.45 + 1.9) (8.14.0.9) ^ - .......... I - - - jj

23 DK 155740 B

TABLE IX

«

PD50 (my / kcj) against I

Compound Staphylococcus, aureus 01Λ167 po sc ampicillin> 100> 100

Compound B> 100> 100 ampicillin + (21.3 + 3.0) +, (16.3 + 1.9) +

Compound B j (21.3 ± 3.0) (16.3 ± 1.9)

. ·· '. ·.: *. ·· ..... TABLE X

pd50 throttleAg) against I

Compound Staphylococcus aureus 01A178 po sc ampicillin> 100> 100

Compound B> 100> 100 ampicillin + (30.0 ± 9.9) + (13.2 ± 2.3) +

Compound B (30.0 ± 9r9) (13.2 ± 2? 3) 24

DK 15S740B

Experiment D

The PD5 o values of ampicillin, compounds A, B and E and 1: 1 mixtures of ampicillin with each of compounds A, B and E, against Staphylococcus aureus 01A400, were measured in mice. The PD ^ values were measured using the procedure described in Experiment C. All doses of ampicillin, compounds A, B and E, and the mixtures thereof, were administered by oral route. The results are listed in Table XI.

TABLE XI

Compound 50 (mg / kg) ampicillin> 100

Compound A> 100

Compound B> 100

Compound E> 100 ampicillin +

Compound A (30.1 ± 2.3) + (30.1 ± 2.3) ampicillin +

Compound B (22.0 ± 1.7) +. (22.0 ± 1.7) ampicillin +

Compound E (54.8 ± 5.1) + (54.8 ± 5.1)

Try

Compound F was orally administered to mice at a single dose of 50 and 20 mg / kg. Blood samples were taken on the mice at intervals starting at 15 minutes after dosing. The blood was allowed to clot and the serum was collected. The serum samples were tested for antibacterial activity using the Comamonas terrigena microorganism. The results are given in Table XII, in which the antibacterial effects are expressed as the concentration of compound A in µg / ml.

DK 155740 B

TABLE XII

Sampling time Antibacterial action in blood (min) (as ng / ml of compound A) 50 mg / kg 20 mg / kg 15 21; 7 ± 3.28 16.0 ± 2.37 30 16.3 ± 2.38 9, 34 + 1.21 45 12.1 ± 1.53 5.80 ± 1.09 60 8.78 ± 0.81 3.58 + 0.81 90 5.34 ± 0.64 1.97 ± 0.63 120 3.13 ± 0.47 0.87 + 0.33 180 1.94 + 0.55 0

26 DK 155740 B

The compounds of the formula I according to the invention increase the antibacterial efficacy of 3-lactam antibiotics in vivo. That is, they reduce the amount of antibiotic needed to protect mice from an otherwise deadly graft of certain 3-lactamase-producing bacteria.

The ability of the compounds of formula I according to the invention to increase the effectiveness of a β-lactam antibiotic against 3-lactamase-producing bacteria makes them valuable for administration with 3-lactam antibiotics in the treatment of bacterial infections in mammals, especially humans. . For the treatment of a bacterial infection, said compound of formula I can be mixed with the 3-lactam antibiotic and the two compounds thereafter administered simultaneously. Alternatively, said compound of formula I may be administered as a separate agent during a course of treatment with a β-lactam antibiotic. In some cases, it will be advantageous to administer the subject compound of formula I beforehand before starting treatment with a 3-lactam antibiotic.

When using penicillanic acid 1,1-dioxide or an in vivo readily hydrolyzable ester thereof to enhance the effectiveness of a 3-lactam antibiotic, the compound is preferably administered in admixture with conventional pharmaceutical carriers or diluents. The formulation methods mentioned previously for use with penicillanic acid 1, 1-dioxide or an in vivo light hydrolyzable ester thereof as a separate antibacterial agent may be used when contemplating administration with another 3-lactam antibiotic. A pharmaceutical composition comprising a pharmaceutically acceptable carrier, a 3-lactam antibiotic and penicillanic acid 1,1-dioxide or a lightly hydrolyzed ester thereof will normally contain from 5 to about 80% by weight of the pharmaceutically acceptable bar.

When penicillanic acid 1,1-dioxide or an in vivo readily hydrolyzable ester thereof is used in combination with another 3-lactam antibiotic, the sulfone may be administered orally or parenterally, i.e. intramuscularly, subcutaneously or intraperitoneally.

Typical 3-lactam antibiotics together with which penicillanic acid-1,1-dioxide and its in vivo easily hydrolyzable esters can be administered are:

27 DK 155740 B

6- (2-phenylacetamido) penicillanic acid, 6- (2-phenoxyacetamido) penicillanic acid, 6- (2-phenylpropipnamido) penicillanic acid. 6- (D-2-Amino-2-phenylacetamoxide) penicillanic acid, 6- (D-2 -amino-2- [4-hydroxyphenyl] acetamido) penicillanic acid, 6- (D-2-amino-2- [1,4-cyclohexadienyl] acetamido) penicillanic acid, 6- (1-aminocyclohexanecarboxamido) penicillanic acid, 6- (2- carboxy-2-phenylacetamido) penicillanic acid, 6- (2-carboxy-2- [3-thienyl] acetamido) penicillanic acid, 6- (D-2- [4-ethylpiperazine-2,3-dione-1-carboxamido] -2 -phenylacetamido) -penicillanic acid, 6- (D-2- [4-hydroxy-1,5-naphthyridine-3-carboxamido] -2-phenylacetamido) -penicillanic acid, 6- (D-2-sulfo-2-phenylacetamido) penicillanic acid , 6- (D-2-sulfoamino-2-phenylac etamido) penicillanic acid, 6- (D-2- [imidazolidin-2-one-1-carboxamido] -2-phenylcetacetamido) penicillanic acid, 6- (D- [3- methylsulfonylimidazolidin-2-one-1-carboxamido] -2-phenylacetamido) penicillanic acid, 6 - ([hexahydro-1H-azepin-1-yl] methyleneamino) penicillanic acid, acetoxymethyl-6- (2-phenylacetamido) penicillanate, acetoxymethane hyl 6- (D-2-amino-2-phenylacetamido) penicillanate, acetoxymethyl 6- (D-2-amino-2- [4-hydroxyphenyl) acetamido) penicillanate, pivaloyloxymethyl 1-6- (2-phenylacetamido) ) penicillanate, pivaloyloxymethyl 6- (D-2-amino-2-phenylacetamido) penicillanate, pivaloyloxymethyl 6- (D-2-amino-2- [4-hydroxyphenyl] acetamido) pyranylate, 1- (ethoxycarhonyloxy) ethyl -6- (2-phenylacetamido) penicillanate, 1- (ethoxycarbonyloxy) ethyl-6- (D-2-amino-2-phenylacetamido) penicillanate, 1- (ethoxycarbonyloxy) ethyl-6- (D-2-amino) 2- [4-hydroxyphenyl] acetamido) -penicillanate, 3-phthalidyl-6- (2-phenylacetamido) penicillanate, 3-phthalidyl-6- (D-2-amino-2-phenylacetamido) penicillanate, 3-phthalidyl-6 (D-2-amino-2- [4-hydroxyphenyl] acetamido) penicillanate, 6- (2-phenoxycarbonyl-2-phenylacetamido) penicillanic acid, 6- (2-tolyloxycarbonyl-2-phenylacetamido) penicillanic acid, 6- (2- 5-indanyloxycarbonyl] -2-phenylacetamido) pencillanic acid, 6- (2-phenoxycarbonyl-2- [3-thienyl] acetamido) penicillanic acid, 6- (2-tholyloxycarbony1-2- [3-thienyl] acetamido) penicill ansyre,

28 DK 155740B

6- (2- [5-indanyloxycarbonyl] -2- [3-thienyl] acetamido) penicillanic acid.

6- (2,2-dimethyl-5-oxo-4-phenyl-1-imidazolidinyl) penicillanic acid, 7- (2- [2-thienyl] acetamido) cephalosporic acid, 7- (2- [1-tetrazolyl] acetamido-3- (2- [5-methyl-1,3,4-thiadiazolyl] thiomethyl) -3-dacetoxymethylcephalosporic acid, 7- (D-2-amino-2-phenylacetamido) desacetoxycephalosporanoic acid, 7-a-methoxy -7 - (2- [2-thienyl] acetamido) -3-carbamoyloxymethyl-3-desacetoxymethylcephalosporic acid, 7- (2-cyanoacetamido) cephalosporanoic acid, 7- (D-2-hydroxy-2-phenylacetamido) -3- (5) - [(1-methyltetrazolyl) thiomethyl) -3-desacetoxymethylcephalosporanoic acid, 7- (2- [4-pyridylthio] acetamido) cephalosporanoic acid, 7- (D-2-amino-2- [1,4-cyclohexadienyl) acetamido) cephalosporanoic acid, 7 - (D-2-amino-2-phenylacetamido) cephalosporanoic acid, and pharmaceutically acceptable salts thereof.

The process according to the invention is further elucidated by the following examples. The IR spectra were measured on KBr slices or Nujol emulsions and the diagnostic absorption bands are indicated as wave numbers (cm 2). The NMR spectra were measured at 60 MHz for solutions in deuterochloroform (CDCl ^), perdeuterodimethylsulfoxide (DMSO-dg) or deuterium oxide (D20), and the peak values are expressed in parts per minute. million (ppm) relative to tetramethylsilane or sodium 2,2-diraethyl-2-silapentane-5-sulfonate. The following abbreviations for tip forms are used: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet.

Example 1

Penicillanic acid 1,1-dioxide.

To a solution of 6.51 g (41 mmol) of potassium permanganate in 130 ml of water and 4.95 ml of glacial acetic acid, cooled to ca. 5 ° C, a cold (about 5 ° C) solution of 4.58 g (21 mmol) of the sodium salt of penicillanic acid in 50 ml of water was added. The mixture was stirred at ca. 5 ° C for 20 minutes, after which the cooling bath was removed. Solid sodium hydrogen sulfite was added until the color of the potassium permanganate had disappeared and then the mixture was filtered. To half of its volume of saturated sodium chloride solution was added to the aqueous filtrate, and then the pH was adjusted to 1.7. The acidic solution was extracted with ethyl acetate. The extracts were dried and then evaporated in vacuo to give 3.47 g of the title product. The aqueous mother liquor was saturated with sodium chloride and extracted further

DK 155740B

more similar to ethyl acetate. The ethyl acetate solution was dried and evaporated in vacuo to give an additional 0.28 g of product. The total yield was then 3.75 g (78% yield). The NMR spectrum (DMSO-dg) of the product showed absorptions at 1.40 (s, 3H), 1.50 (s, 3H), 3.13 (d of d's, 1H, J ± = 16Hz, J2 = 2Hz) , 3.63 (d, of d's, 1H, J1 = 16 Hz, J2 = 4Hz), 4.22 (s, 1H) and 5.03 (d of d's, 1H, J · 4 = 4Hz, J2 = 2Hz ) ppm.

Example 2

Penicillanic acid 1,1-dioxide.

a) Benzylpenicillanate-1,1-dioxide.

To a stirred solution of 6.85 g (24 mmol) of benzylpenicillate in 75 ml of ethanol-free chloroform under nitrogen in an ice bath was added in 2 portions, at several intervals, 4.78 g of 85% pure 3-chloro-perbenzoic acid. Stirring was continued for 30 minutes in the ice bath and then for 45 minutes without external cooling. The reaction mixture was washed with aqueous alkali (pH 8.5) followed by saturated sodium chloride solution and then dried and evaporated in vacuo to give 7.05 g of residue. Examination of this residue showed that it was a 5.5: 1 mixture of benzylpenicillanate-1-oxide and benzylpenicillanate-1,1-dioxide.

To a stirred solution of 4.85 g of the above 5.5: 1 sulfoxide-sulfone mixture in 50 ml of ethanol-free chloroform under nitrogen was added 3.2 g of 85% pure 3-chloroperbenzoic acid at room temperature. The reaction mixture was stirred for 2.5 hours, then diluted with ethyl acetate. The resulting mixture was added to water at pH 8.0 and then the layers were separated. The organic phase was washed with water of pH 8.0 followed by saturated sodium chloride solution and then dried using sodium sulfate. Evaporation of the solvent in vacuo yielded 3.59 g of the title compound. The NMR spectrum of the product (in CDCl 3) showed absorptions at 1.28 (s, 3H), 1.58 (s, 3H), 3.42 (m, 2H), 4.37 (s, 1H), 4, 55 (m, 1H), 5.18 (q, 2H, J = 12 Hz) and 7.35 (s, 5H) ppm.

b) Penicillanic acid 1,1-dioxide.

To a stirred solution of 8.27 g of benzylpenicillanate-1,1-dioxide in a mixture of 40 ml of methanol and 10 ml of ethyl acetate was slowly added 10 ml of water followed by 12 g of 5% palladium-on-calcium carbonate. The mixture was shaken under a hydrogen atmosphere at 3.7 ° C for 40 minutes.

DK 155740B

30 nuts, after which it was filtered through supercell (a diatomaceous earth). The filter cake was washed with methanol and aqueous methanol and the washings were added to the filtrate. The combined solution was evaporated in vacuo to remove the bulk of the organic solvents and then the residue was partitioned between ethyl acetate and water at a pH of 2.8. The ethyl acetate layer was removed and the aqueous phase was further extracted with ethyl acetate. The combined ethyl acetate solutions were washed with saturated sodium chloride solution, dried using sodium sulfate and then evaporated in vacuo. The residue is slurried in a 1: 2 mixture of ethyl acetate and ether to give 2.37 g of the title product, m.p. 148-151 ° C. The ethyl acetate-ether mixture was evaporated to give an additional 2.17 g of product.

Example 3

Pivaloyloxymethylpenicillanat-1,1-dioxide.

To 0.562 g (2.41 mmol) of penicillanic acid 1,1-dioxide in 2 ml of N, N-dimethylformamide was added 0.375 g (2.90 mmol) of diisopropylethylamine followed by 0.360 ml of chloromethyl pivalate. The reaction mixture was stirred at room temperature for 24 hours, then diluted with ethyl acetate and water. The ethyl acetate layer was separated and washed 3 times with caustic and 1 time with saturated sodium chloride solution. The ethyl acetate solution was then dried using anhydrous sodium sulfate and evaporated in vacuo to give 0.700 g of the title product as a solid, m.p. 103-104 ° C. The NMR spectrum of the product (in CDCl3) showed absorptions at 1.27 (s, 9H), 1.47 (s, 3H), 1.62 (s, 3H), 3.52 (m, 2H), 4 , 47 (s, 1H), 4.70 (m, 1H), 5.73 (d, 1H, J = 6.0 Hz) and 5.98 (d, 1H, J = 6.0 Hz).

Example 4

Acetoxymethylpenic illanate-1,1-dioxide.

The procedure of Example 3 was repeated except that the pivaloyloxymethyl chloride used therein was replaced with an equimolar amount of acetoxymethyl chloride. This yielded 56% yield of acetoxymethylpenicillanate-1,1-dioxide, m.p. 138-141 ° C. The NMR spectrum of the product (in CDCl3) showed absorptions at 1.45 (s, 3H), 1.65 (s, 3H), 2.10 (s, 3H), 3.45 (d, 2H, J = 3 Hz), 4.40 (s, 1H), 4.50 (t, 1H, J = 3 Hz) and 5.75 (q, 2H, J = 5 Hz) ppm down the field from internal tetramethylsilane.

31 DK 155740B

Acetoxymethylpenicillanate-1,1-dioxide was also prepared in 29% yield from sodium penicillanate-1,1-dioxide and acetylmethyl bromide, in dimethylsulfoxide, in a similar manner to Example 3. After chromatography, the product had a melting point of 143-144. ° C. Its IR spectrum in a KBr disk showed absorptions at 1750, 1300, 1150, 1040, 990 and 825 cm @ 1 of the NMR spectrum of DMSO-dg showed absorptions at 1.00 (s, 3H), 1.15 (s, 3H), 1.75 (s, 3H), 2.95 (q, 1H), 3.15 (q, 1H), 4.05 (s, 1H), 4.80 (q, 1H), and 5, 40 (q, 2H) ppm down in the field from internal tetramethylsilane.

Example 5

Methoxycarbonyloxymethylpenicillanat-1,1-dioxide.

A rapid flow of chlorine gas was bubbled through a stirred solution of 3.08 ml of methyl chloroformate in 50 ml of deoxygenated carbon tetrachloride under nitrogen. This reaction mixture was then irradiated with light at a wavelength of 3500 Å for 10 minutes. The carbon tetrachloride solution was then cooled to -10 ° C and 1.62 ml of methanol was added. Then, 11.87 ml of diisopropylethylamine was added dropwise in 10 ml of carbon tetrachloride. The reaction mixture was allowed to warm slowly to room temperature and stirring was continued for 45 minutes. The solvent was removed by evaporation in vacuo and then a solution of 6.24 g of penicillanic acid 1,1-dioxide and 4.62 ml of diisopropylethylamine in 60 ml of Ν, Ν-dimethylformamide was added. Stirring was continued for 24 hours and then water and dichloromethane were added. The pH was adjusted to 4.0 and the layers separated. The aqueous phase was further extracted with dichloromethane and then the combined dichloromethane layers were washed with water of pH 3.0 followed by water without any pH adjustment. The dried dichloromethane solution was evaporated in vacuo to an oil which solidified by ether treatment. There was thus obtained 1.94 g of the title compound, m.p. 124-126 ° C. The NMR spectrum (CDCl3) showed absorptions at 1.43 (s, 3H), 1.61 (s, 3H), 3.44 (m, 2H), 3.85 (s, 3H), 4.39 (s). , 1H), 4.59 (m, 1H) and 5.78 (q, 2H) ppm down in the field from tetramethylsilane.

32 DK 155740 B

Example 6

Butoxycarbonyloxymethylpenicillanat-1,1-dioxide.

A rapid stream of chlorine gas was bubbled through a stirred solution of 3.08 ml of methyl chloroformate in 50 ml of deoxygenated carbon tetrachloride. This reaction mixture was then irradiated with light at a wavelength of 3500 Å for 7 minutes. The reaction mixture was purged with nitrogen and then irradiated for a further 10 seconds. The solution thus obtained was cooled to ca. -10 ° C, then 3.48 ml of n-butanol was added. Then 8.6 ml of diisopropylethylamine was added dropwise in 10 ml of dichloromethane. The cooling bath was removed and stirring was continued for 1 hour. The dichloromethane was removed in vacuo and then a solution of 3.07 g of penicillanic acid 1,1-dioxide and 2.27 ml of diisopropylethylamine was added in ca. 50 ml of Ν, Ν-dimethylformamide. Stirring was continued for 18 hours and then water and dichloromethane were added. The layers were separated and the aqueous phase was further extracted with dichloromethane. The combined dichloromethane solutions were washed with water, dried and evaporated to give the crude product.

Analysis of the crude product by NMR spectroscopy revealed that the reaction was incomplete. Accordingly, the crude product was dissolved in 5 ml of Ν, Ν-dimethylformamide and 1.165 g of penicillanic acid-1,1-dioxide and 0.86 ml of diisopropylethylamine were added. This mixture was stirred overnight and the product was then isolated as before. The reaction was still incomplete and therefore the product was redissolved in 5 ml of Ν, Ν-dimethylformamide, and 1.165 g of penicillanic acid 1,1-dioxide and 0.86 ml of diisopropylethylamine were added. The mixture was stirred overnight and the product isolated as before. 4.6 g of crude product were obtained.

The latter crude product was purified by column chromatography on silica gel to give 0.19 g of the title compound. The NMR spectrum (CDCl3) showed absorptions at 0.98 (t, 3H), 1.45 (s, 3H), 1.63 (s, 3H), 1.55 (m, 4H), 3.5 ( m, 2H), 4.23 (t, 3H, J = 6.8 Hz), 4.45 (s, 1H), 4.65 (m, 1H) and 5.85 (q, 2H) ppm downwards the field from tetramethylsilane.

33 DK 1S5740B

Example 7 3-Phthalidylpenicillanate-1,1-dioxide.

To 0.783 g (3.36 mmol) of penicillanic acid 1,1-dioxide in 5 ml of N, N-dimethylformamide was added 0.47 ml of triethylamine, followed by 0.715 g of 3-bromophthalide. The reaction mixture was stirred for 2 hours at room temperature, then diluted with ethyl acetate and water, the pH of the aqueous phase was raised to 7.0 and the layers separated. The ethyl acetate layer was washed successively with water and saturated sodium chloride solution, then dried using sodium sulfate. The ethyl acetate solution was evaporated in vacuo to give the title product as a white foam. The NMR spectrum of the product (in CDCl 3) showed absorptions at 1.47 (s, 6H), 3.43 (m, 1H), 4.45 (s, 1H), 4.62 (m, 1H), 7, 40 and 7.47 (2s's, 1H) and 7.73 (m, 4H) ppm.

By repeating the above procedure, except that the 3-bromophthalide was replaced by 4-bromo-crotonolactone and 4-bromo-Y-butyrolactone, respectively: 4-crotonolactonylpenicillanate-1,1-dioxide and Y-butyrolacton-4-yl-1 -penicillanate.

Example 8 1- (Ethoxycarbonyloxy) ethylpenicillanate-1,1-dioxide.

A mixture of 0.654 g of penicillanic acid 1,1-dioxide, 0.42 ml of triethylamine, 0.412 g of 1-chloroethyl-ethyl carbonate, 0.300 g of sodium bromide and 3 ml of Ν, Ν-dimethylformamide was stirred at room temperature for 6 days. The mixture was then worked up by dilution with ethyl acetate and water and then adjusted to pH 8.5. The ethyl acetate layer was separated and washed 3 times with water and 1 time with saturated sodium chloride solution, then dried using anhydrous sodium sulfate. The ethyl acetate was removed by evaporation in vacuo to give 0.390 g of the title product as an oil.

The above product was poured together with an approximately equal amount of a similar material from a similar experiment. The combined product was dissolved in chloroform and 1 ml of pyridine was added. The mixture was stirred at room temperature overnight, then the chloroform was removed by evaporation in vacuo. The residue was partitioned between ethyl acetate and water at pH 8. The separated and dried ethyl acetate was then evaporated in vacuo to give 150 mg of the title product (yield, about 7%). The IR spectrum (film) of the product showed absorptions at 1805 and 1763 cm NMR spectrum (CDCl

34 DK 155740B

showed absorptions at 1.43 (m, 12H), 3.47 (m, 2H), 3.9 (q, 2H, J = 7.5 Hz), 4.37 / m, 1H), 4.63 ( m, 1H) and 6.77 (m, 1H) ppm.

Example 9

The procedure of Example 8 was repeated except that the 1-chloroethyl ethyl carbonate was replaced by an equimolar amount of the 1-chloroalkyl alkyl carbonate required, 1- (alkanoyloxy) ethyl chloride or 1-methyl (alkanoyloxy) ethyl chloride to give the following compounds, respectively. : methoxycarbonyloxymethylpenicillanate-1,1-dioxide, ethoxycarbonyloxymethylpenicillanate-1,1-dioxide, isobutoxycarbonyloxymethylpenicillanate-1,1-dioxide, 1- (methoxycarbonyloxy) ethylpenicillanate-1,1-dioxide, 1- (butoxycarbonyloxy-dioxide) , 1- (acetoxy) ethylpenicillanate-1,1-dioxide, 1- (butyryloxy) ethylpenicillanate-1,1-dioxide, 1- (pivaloyloxy) ethylpenicillanate-1,1-dioxide, 1- (hexanoyloxy) ethylpenicillanate-1,1 -dioxide, 1-methyl-1- (acetoxy) ethylpenicillanate-1,1-dioxide and 1-methyl-1- (isobutyryloxy) ethylpenicillanate-1,1-dioxide.

Example 10

Penicillanic acid 1,1-dioxide.

To 2.17 g (10 mmol) of penicillanic acid 1-oxide in 30 ml of ethanol-free chloroform at ca. At 0 ° C was added 1.73 g (10 mmol) of 3-chloroperbenzoic acid. The mixture is stirred for 1 hour at ca. 0 ° C and then for another 24 hours at 25 ° C. The filtered reaction mixture is evaporated in vacuo to give penicillanic acid 1,1-dioxide. Mp. 148-151 C.

Example 11

The procedure of Example 10 was repeated except that the penicillanic acid-la-oxide used therein was replaced with: penicillanic acid-l3-oxide, acetoxymethylpenicillanate-la-oxide, propionyloxymethylpenicillanate-la-oxide, pivaloyloxymethyl-epicillan-oxide-1 oxide,

DK 155740 B

propionyloxymethylpenicillanate-oxide, pivaloyloxymethylpenicillanate-13-oxide, 3-phthalidylpenicillanate-1-oxide, 3-phthalidylpenicillanate-13-oxide, 1- (ethoxycarbonyloxy) illanate ΐα-oxide, 1- (methoxycarbonyloxy) ethylpenicillanate-la-oxide, 1- (butoxycarbonyloxy) ethylpenicillanate-la-oxide, 1- (acetoxy) ethylpenicillanate-la-oxide, 1- (butyryloxy) ethylpenicillanate-la-oxide, 1- (pivaloyloxy) ethylpenicillanate 1-oxide, 1- (ethoxycarbonyloxy) ethylpenicillanate-13-oxide, methoxycarbonyloxymethylpenicillanate-10-oxide, ethoxycarbonyloxymethylpenicillanate-13-oxide, isobutoxycarbonyloxymethylpenicillanate-oxide , 1- (butoxycarbonyloxy) ethylpenicillanate ip oxide, 1- (acetoxy) ethylpenicillanate 13 oxide, 1- (butyryloxy) ethylpenicillanate 13 oxide and 1- (pivaloyloxy) ethylpenicillanate 13 oxide.

There were obtained respectively: penicillanic acid 1,1-dioxide, acetoxymethylpenicillanate-1,1-dioxide, m.p. 143-144 ° C, propionyloxymethylpenicillanate-1,1-dioxide, pivaloyloxymethylpenicillanate-1,1-dioxide, m.p. 103-104 ° C, acetoxymethylpenicillanate-1,1-dioxide, propionyloxymethylpenicillanate-1,1-dioxide, pivaloyloxymethylpenicillanate-1,1-dioxide, 3-phthalidylpenicillanate-1,1-dioxide, 3-phthalidylpenicillanate-1,1-dioxide v 1- (ethoxycarbonyloxy) ethylpenicillanate-1,1-dioxide, methoxycarbonyloxymethylpenicillanate-1,1-dioxide, m.p. 124-126 ° C, ethoxycarbonyloxymethylpenicillanate-1,1-dioxide, isobutoxycarbonyloxymethylpenicillanate-1, 1-dioxide, 1- (methoxycarbonyloxy) ethylpenicillanate-1,1-dioxide, 1- (butoxycarbonyloxy) ethylpenicillanate-1,1-dioxide - (acetoxy) ethylpenicillanat-1,1-dioxide,

36 DK 155740B

1- (butyryloxy) ethylpenicillanate-1,1-dioxide, 1- (pivaloyloxy) ethylpenicillanate-1,1-dioxide, 1- (ethoxycarbonyloxy) ethylpenicillanate-1,1-dioxide, methoxycarbonyloxymethylpenicillanate-1,1-dioxide, ethoxycarbonyloxymethylpenyl , 1-dioxide, isobutoxycarbonyloxymethylpenicillanate-1,1-dioxide, 1- (methoxycarbonyloxy) ethylpenicillanate-1,1-dioxide, 1- (butoxycarbonyloxy) ethylpenicillanate-1,1-dioxide, 1- (acetoxy) ethylpenicillanate-1,1-dioxide dioxide, 1- (butyryloxy) ethylpenicillanate-1,1-dioxide and 1- (pivaloyloxy) ethylpenicillanate-1,1-dioxide.

Example 12

Penicillanic acid 1,1-dioxide.

Similarly, by oxidation of 4-nitrobenzyl-penicillanate-1-oxide and 4-nitrobenzyl-penicillanate-1-oxide with 3-chloroperbenzoic acid, the method of Example 10 obtained 4-nitro-benzyl-penicillanate-1,1-dioxide.

Hydrogenolysis of 4-nitrobenzylpenicillanate-1,1-dioxide by the method of Example 2b) gives penicillanic acid 1,1-dioxide.

Example 13

Of sodium penicillanate 1,1-dioxide.

To a stirred solution of 32.75 g (0.14 mmol) of penicillanic acid 1,1-dioxide in 450 ml of ethyl acetate was added a solution of 25.7 g of sodium 2-ethylhexanoate (0.155 mol) in 200 ml of ethyl acetate. The resulting solution was stirred for 1 hour, then an additional 10% excess of sodium 2-ethyl hexanoate was added in a small volume of ethyl acetate. The product immediately began to precipitate. Stirring was continued for 30 minutes, and then the precipitate was diluted by filtration. It was washed successively with ethyl acetate, with 1: 1 ethyl acetate ether and with ether. The solid was then dried over phosphorus pentoxide at ca. 0.1 mm Hg for 16 hours at 25 ° C to give 36.8 g of the title sodium salt contaminated with a small amount of ethyl acetate. The ethyl acetate content was reduced by heating to 100 ° C for 3 hours in vacuo. The IR spectrum of this final product (KBr disk) showed absorptions at 1786 and 1608 cm ’’. The NMR spectrum (D20) showed the absorbance.

37 DK 155740 B

tions at 1.48 (s, 3H), 1.62 (s, 3H), 3.35 (d of d's, 1H, J ^ = 16Hz, J2 = 2Hz), 3.70 (d of d's, 1H, = 16 =, J₂ = 4Hz), 4.25 (s, 1H) and 5.03 (d of d's, 1H, J J = 4Hz, J₂ = 2Hz) ppm.

The titled sodium salt may also be prepared using acetone instead of ethyl acetate.

Example 14

Penicillanic acid 1,1-dioxide.

To a mixture of 7600 ml of water and 289 ml of glacial acetic acid was added portionwise 379.5 g of potassium permanganate. This mixture was stirred for 15 minutes and then cooled to 0 ° C. To the mixture was then added, with stirring, a mixture which had been prepared from 270 g of penicillanic acid, 260 ml of 4 N sodium hydroxide solution and 2400 ml of water (pH 7.2), which had then been cooled to 8 ° C. The temperature rose to 15 ° C during this last addition. The temperature of the mixture obtained was lowered to 5 ° C and stirring was continued for 30 minutes. To the reaction mixture was then added 142.1 g of sodium hydrogen sulfite in portions over 10 minutes. The mixture was stirred for 10 minutes at 10 ° C and then 100 g of supercell (a diatomaceous earth) was added. After a further 5 minutes of stirring, the mixture was filtered. To the filtrate was added 4.0 liters of ethyl acetate and then the pH of the aqueous phase was lowered to 1.55 using 6N hydrochloric acid. The ethyl acetate layer was removed and poured with several additional ethyl acetate extracts. The combined organic layer was washed with water, dried (MgSO 4) and evaporated almost to dryness in vacuo. The slurry thus obtained was stirred with 700 ml of ether at 10 ° C for 20 minutes, after which the solid was collected by filtration. Thereby 82.6 g (26% yield) of the title compound was obtained with m.p. 154-155.5 ° C (dec.).

Example 15

Pivaloyloxymethylpenicillanat-1,1-dioxide.

To a solution of 1.25 g of pivaloyloxymethylpenicillanate in 40 ml of chloroform, cooled to ca. -15 ° C, 0.8 g of 3-chloroperbenzoic acid was added. The mixture was stirred at ca. -15 ° C for 20 minutes, then allowed to warm to room temperature. Analysis of the obtained solution by NMR showed that it contained both the 1α and 10 oxide.

38 DK 155740 B

The chloroform solution was concentrated to ca. 20 ml and a further 0.8 g of 3-chloroperbenzoic acid was added. The resulting mixture was stirred overnight at room temperature, after which all the solvent was removed by evaporation in vacuo. The residue was dissolved again for approx. 4 ml of dichloromethane and 0.4 g of 3-chlorobenzoic acid were added. The mixture was stirred for 3 hours, after which the solvent was removed by evaporation in vacuo. The residue was partitioned between ethyl acetate and water at pH 6.0, and sodium hydrogen sulfite was added until a test for the presence of peroxides was negative. The pH of the aqueous phase was raised to 8.0 and the layers separated. The organic layer was washed with brine, dried using anhydrous sodium sulfate and evaporated in vacuo. The residue was dissolved in ether and precipitated again by the addition of hexane. The solid obtained was recrystallized from ether to give 0.357 g of the title compound.

The NMR spectrum of the product (CDCl 3) showed absorptions at 1.23 (s, 9H), 1.50 (s, 3H), 1.67 (s, 3H), 3.28 (m, 2H), 4.45 (s, 1H), 5.25 (m, 1H) and 5.78 (m, 2H) ppm.

Example 16 3 3-Phthalidylpenicillanate-1,1-dioxide.

To a solution of 713 mg of 3-phthalidylpenicillanate in 3 ml of chloroform was added 0.430 g of 3-chloroperbenzoic acid at ca. 10 ° C. The mixture was stirred for 30 minutes, then an additional 0.513 g of 3-chloroperbenzoic acid was added. The mixture was stirred for 4 hours at room temperature, after which the solvent was removed by evaporation in vacuo. The residue was partitioned between ethyl acetate and water at pH 6.0 and sodium hydrogen sulfite was added to decompose the residual per acid. The pH of the aqueous phase was boiled to 8.8. The layers were separated and the organic phase was evaporated in vacuo. Thereby the title compound was obtained as a foam. The NMR spectrum (CDCl3) showed absorptions at 1.62 (m, 6H), 3.3 (m, 2H), 4.52 (p, 1H), 5.23 (m, 1H) and 7.63 ( m, 5H) ppm.

39 DK 155740 B

Example 17

Penicillanic acid 1,1-dioxide.

a) 4-Nitrobenzylpenicillanate-1,1-dioxide.

A solution of 4-nitrobenzylpenicillanate in chloroform was cooled to ca. 15 ° C and 1 equivalent of 3-chloroperbenzoic acid was added. The reaction mixture was stirred for 20 minutes. An examination of the reaction mixture at this time by NMR spectroscopy revealed that it contained 4-nitrobenzylpenicillanate 1-oxide. An additional 1 equivalent of 3-chloroperbenzoic acid was added and the reaction mixture was stirred for 4 hours. At this time, an additional 1 equivalent of 3-chloroperbenzoic acid was added and the reaction mixture was stirred overnight. The solvent was removed by evaporation and the residue partitioned between ethyl acetate and water at pH 8.5. The ethyl acetate layer was separated, washed with water, dried and evaporated to give the crude product. The crude product was purified by chromatography on silica gel eluting with a 1: 1 mixture of ethyl acetate and chloroform.

The NMR spectrum of this product (CDCl3) showed absorptions at 1.35 (s, 3H), 1.58 (s, 3H), 3.45 (m, 2H), 4.42 (s, 1H), 4, 58 (m, 1H), 5.30 (s, 2H) and 7.83 (q, 4H) ppm.

b) Penicillanic acid 1,1-dioxide.

To 0.54 g of 4-nitrobenzylpenicillanate-1,1-dioxide in 30 ml of methanol and 10 ml of ethyl acetate was added 0.54 g of 10% palladium-on-carbon. The mixture was then shaken under a hydrogen atmosphere at a pressure of approx. 3.5 at until hydrogen uptake ceased. The reaction mixture was filtered and the solvent removed by evaporation. The residue was partitioned between the ethyl acetate and water at pH 8.5 and the aqueous layer removed. Fresh ethyl acetate was added and the pH was adjusted to 1.5. The ethyl acetate layer was removed, washed with water and dried, and then evaporated in vacuo. There was thus obtained 0.168 g of the title compound as a crystalline solid.

40

DK 155740B

c) Penicillanic acid 1,1-dioxide.

A stirred solution of 512 mg of 4-nitrobenzylpenicillanate-1,1-dioxide in a mixture of 5 ml of acetonitrile and 5 ml of water was cooled to 0 ° C and then a solution of 484 mg of sodium dithionite was added portionwise in 1.4 ml. 1.0 N sodium hydroxide over several minutes. The reaction mixture was stirred for a further 5 minutes and then diluted with ethyl acetate and water with pH 8.5. The ethyl acetate layer was separated and evaporated in vacuo to give 300 mg of starting material. Fresh ethyl acetate was added to the aqueous phase and the pH was adjusted to 1.5. The ethyl acetate was separated, dried and evaporated in vacuo to give 50 mg of the title compound.

Example 18 1-Methyl-1- (acetoxy) ethylpenicillanate-1,1-dioxide.

To 2.33 g of penicillanic acid 1,1-dioxide in 5 ml of Ν, Ν-dimethylformamide was added 1.9 ml of ethyl diisopropylamine followed by dropwise addition of 1.37 g of 1-methyl-1- (acetoxy) ethyl chloride at about 20 °. c. The mixture was stirred at room temperature overnight, then quenched with ethyl acetate and m 2 of water. The layers were separated and the ethyl acetate layer was washed with water of pH 9. The ethyl acetate solution was then dried (Na 2 SO 4) and evaporated in vacuo to give 1.65 g of crude product as an oil. The oil solidified upon standing in the refrigerator and the product was then recrystallized from a mixture of chloroform and ether to give a product of m.p. 90-92 ° C.

The NMR spectrum of the crude product (CDCl 3) showed absorptions at 1.5 (s, 3H), 1.62 (s, 3H), 1.85 (s, 3H), 1.93 (s, 3H), 2 , 07 (s, 3H), 3.43 (m, 2H), 4.3 (s, 1H) and 4.57 (m, 1H) ppm.

Example 1 g

The procedure of Example 18 was repeated except that the 1-methyl-1 (acetoxy) ethyl chloride was replaced with the necessary 1-methyl-1- (alkanoyloxy) ethyl chloride to give the following compounds, respectively: 1-methyl-1- (propionyloxy) ethylpenicillanate -1,1-dioxide, 1-methyl-1- (pivaloyloxy) ethylpenicillanate-1,1-dioxide and 1-methyl-1- (hexanoyloxy) ethylpenicillanate-1,1-dioxide.

41 DK 155740 B

Example 20

Penicillanic acid 1,1-dioxide.

To a stirred solution of 1.78 g of penicillanic acid in pH 7.5 water was added 1.46 ml of 40% peracetic acid followed by an additional 2.94 ml of 40% peracetic acid 30 minutes later. The reaction mixture was stirred for 3 days at room temperature, then diluted with ethyl acetate and water. Solid sodium hydrogen sulfite was added to decompose excess peracid, and then the pH was adjusted to 1.5. The ethyl acetate layer was separated, dried (Na 2 O 4) and evaporated in vacuo. The residue was a 3: 2 mixture of penicillanic acid 1,1-dioxide and penicillanic acid 1-oxide.

Example 21

Pivaloyloxymethylpenicillanat-1,1-dioxide.

A stirred solution of 595 mg of pivaloyloxymethylpenicillanate-1 oxide in 5 ml of ethyl acetate was cooled to ca. -15 ° C and 5 mg of manganese acetylacetonate were added. To the dark brown mixture thus obtained, 0.89 ml of 40% peracetic acid was added in small amounts over several minutes over several minutes. After 40 minutes, the cooling bath was removed and the mixture was stirred at room temperature for 3 days. The mixture was diluted with ethyl acetate and water at pH 8.5 and the ethyl acetate layer was separated, dried and evaporated in vacuo. This yielded 178 mg of material which, by NMR spectroscopy, proved to be a mixture of pivaloyloxymethylpenicillanate-1,1-dioxide and piva-loyloxymethylpenicillanate-1 oxide.

The above material was again dissolved in ethyl acetate and re-oxidized using 0.9 ml of peracetic acid and 5 mg of manganese acetylacetonate as described above using a reaction time of 16 hours. The reaction mixture is worked up as described above. There was obtained 186 mg of pivaloyloxymethylpenicillanate-1,1-dioxide.

Reference Example 1

Penicillanic acid la-oxide.

To 1.4 g of pre-hydrogenated 5% palladium-on-calcium carbonate in 50 ml of water was added a solution of 1.39 g of benzyl-6,6-dibrompenicilanate-la-oxide in 50 ml of tetrahydrofuran. The mixture was shaken under a hydrogen atmosphere at ca. 3.2 at and 25 ° C for 1 hour, then filtered. The filtrate was evaporated in vacuo to remove the main I

42 DK 155740 B

the amount of the tetrahydrofuran, after which the aqueous phase is extracted with ether. The ether extracts were evaporated in vacuo to give 0.5 g of material which was found to be mainly benzylpenicilanate la oxide.

The above-mentioned benzylpenicillanate-la-oxide was poured into an additional 2.0 g of benzyl-6,6-dibrompenicillanate-la-oxide and the mixture was dissolved in 50 ml of tetrahydrofuran. The solution was added to 4.0 g of 5% palladium-on-calcium carbonate in 50 ml of water and the resulting mixture was shaken under a hydrogen peroxide at ca. 3.2 at and 25 ° C overnight. The mixture was filtered and the filtrate extracted with ether. The extracts were evaporated in vacuo and the residue purified by chromatography on silica gel eluting with chloroform. 0.50 g of material was obtained.

The latter material was hydrogenated at ca. 3, -2 at and 25 ° C in water-methanol (1: 1) with 0.50 g of 5% palladium-on-calcium carbonate for 2 hours. After this time, an additional 0.50 g of 5% palladium-on-calcium carbonate was added and hydrogenation was continued at 3.2 at and 25 ° C overnight. The reaction mixture was filtered and extracted with ether and the extracts discarded. The remaining aqueous phase was adjusted to pH 1.5 and then extracted with ethyl acetate. The ethyl acetate extracts were dried (Na 2 SO 4) and then evaporated in vacuo to give 0.14 g of penicillanic acid la-oxide.

The NMR spectrum (CDCl3 / DMSO-dg) showed absorptions at 1.4 (s, 3H), 1.64 (s, 3H), 3.60 (m, 2H), 4.3 (s, 1H) and 4.54 (m, 1H) ppm. The IR spectrum of the product (KBr disk) showed absorptions at 1795 and 1745 cm 2

Reference Example 2

Penicillanic acid la-oxide.

To 1.0 g of pre-hydrogenated 5% palladium-on-calcium carbonate in 30 ml of water was added a solution of 1.0 g of 6,6-dibrompenicillanic acid la-oxide. The mixture was shaken under a hydrogen atmosphere at ca. 3.2 at and 25 ° C for 1 hour. The reaction mixture was then filtered and the filtrate was concentrated in vacuo to remove the methanol. The remaining aqueous phase was diluted with an equal volume of water, adjusted to pH 7 and washed with ether. The aqueous phase was then acidified to pH 2 with dilute hydrochloric acid and extracted with ethyl acetate, the ethyl acetate extracts were dried (Na 2 SO 4) and evaporated in vacuo to give penicillanic acid la-oxide.

43 DK 155740 B

Reference Example 3

Penicillanic acid 13-oxide.

To a stirred solution of 2.65 g (12.7 mmol) of penicillanic acid in chloroform at 0 ° C was added 2.58 g of 85% pure 3-chloroperbenzoic acid. After 1 hour, the reaction mixture was filtered and the filtrate was evaporated in vacuo. The residue was dissolved in a small amount of chloroform. The solution was concentrated slowly until a precipitate began to appear. At this point, evaporation was stopped and the mixture was diluted with ether. The precipitate was removed by filtration, washed with ether and dried to give 0.615 g of penicillanic acid ip oxide, m.p. 140-143 ° C. The IR spectrum of the product (CHCl 3 solution) showed ab sorptions at 1775 and 1720 cm. The NMR spectrum (CDCl , 05 (m, 1H) ppm. According to the NMR spectrum, the product was approx. 90% pure.

A study of the chloroform-ether mother liquor showed that it contained additional penicillanic acid-13 oxide and, moreover, some penicil-lanoic acid-la-oxide.

Reference Example 4

By esterification of penicillanic acid la-oxide or penicillanic acid-13-oxide with the required alkanoyloxy chloride in accordance with Example 4, the following compounds were obtained, respectively: acetoxymethylpenicillanate-la-oxide, propionyloxymethylpenicillanate-la-oxide, pivalojzbxymethylpenicillanate-oxide, -13-oxide, propionyloxymethylpenicillanate ip-oxide and pivaloyloxymethylpenicillanate-13-oxide.

Reference Example 5

By reaction of penicillanic acid-la-oxide or penicillanic acid-β-oxide with 3-bromophthalide, 4-bromocrotonolactone or 4-bromo-γ-butyrolactone, respectively, the following compounds were obtained: 3-phthalidylpenicillanate-1-oxide, 4- crotonolactonylpenicillanate 1-oxide, 3-phthalidylpenicillanate-13-oxide, 4-crotonolactonyl-penicillanate-13-oxide and v-hnf wrnl d-nonϊ pi 1 1 anat-1 R-ηνϊ

44 DK 155740B

Reference Example 6

By reaction of penicillanic acid 1a-oxide or penicillanic acid Ιβ-oxide with the required 1-chloroalkyl-alkyl carbonate or 1- (alkanoyloxy) ethyl chloride according to the procedure of Example 8, the following compounds were obtained, respectively: 1- (ethoxycarbonyloxy) ethylpenieillanate -la oxide, methoxycarbonyloxymethylpenicillanate la oxide, ethoxyc arbonyloxymethylpenic illanate la oxide, isobutoxycarbonyloxymethylpenicillanate la oxide, 1- (methoxycarbonyloxy) ethylpenicillanate la oxide, 1- (butoxycarbonyloxy) acetoxy) ethylpenicillanate 1 oxide, 1- (butyryloxy) ethylpenicillanate 1 oxide, 1- (pivaloyloxy) ethylpenicillanate 1 oxide, 1- (ethoxycarbonyloxy) ethylpenicillanate 10 oxide, methoxycarbonyloxymethylpenicillanate 13 oxide, ethoxycarbony oxide, in sobutoxycarbonyloxymethylpenic illanate 13 oxide, 1- (methoxycarbonyloxy) ethylpenicillanate-13 oxide, 1- (butoxycarbonyloxy) ethylpenicillanate-13 "Oxide, 1- (acetoxy) ethylpenicillanate-13 oxide, 1- (butyryloxy) ) Ethylpenicillanate-13 "Oxide and 1- (pivaloyloxy) ethylpenicillanate-13 oxide.

Reference Example 7

By reaction of penicillanic acid-la-oxide and penicillanic acid-13-oxide with benzyl bromide according to the procedure of Example 3, benzylpenicillanate-la-oxide and benzyl-penicillanate-13 ° Oxide were obtained, respectively.

Similarly, by reaction of penicillanic acid 1a-oxide and penicillanic acid-13 ~ oxide with 4-nitrobenzyl bromide, in accordance with the procedure of Example 3, 4-nitro-benzylpenicillanate-1-oxide and 4-nitrobenzylpenicillanate-13-oxide, respectively, were obtained.

Preparation A

6,6-dibromopenicillanic acid la-oxide.

The title compound is prepared by oxidation of 6,6-dibrompenicillanic acid with 1 ash valent 3-chloroperbenzoic acid.

45 DK 155740 B

acid in tetrahydrofuran at 0-25 ° C for approx. 1 hour following the method set forth by Harrison et al., Journal of the Chemical Society (London) Perkin I, 1772 (1976).

Preparation B

Benzyl 6,6-dibromopenicillanate.

To a solution of 54 g (0.165 mol) of 6,6-dibrompenicillanic acid in 350 ml of Ν, Ν-dimethylacetamide was added 22.9 ml (0.165 mol) of triethylamine and the solution was stirred for 40 minutes. 19.6 ml (0.165 mol) of benzyl bromide was added and the resulting mixture was stirred at room temperature for 48 hours. The precipitated triethylamine hydrobromide was filtered off and the filtrate was added to 1500 ml of ice water adjusted to pH 2. The mixture was extracted with ether and the extracts were washed successively with saturated sodium bicarbonate solution, water and brine. The dried (MgSO 4) ether solution was evaporated in vacuo to give an almost white solid which was recrystallized from isopropanol. There was thus obtained 70.0 g (95% yield) of the title compound, m.p. 75-76 ° C. The IR spectrum (KBr disk) showed absorptions at 1795 and 1740 cm NMR spectrum (CDCl 3) showed absorptions at 1.53 (s, 3H), 1.58 (s, 3H), 4.50 (s, 1H) , 5.13 (s, 2H), 5.72 (s, 1H) and 7.37 (s, 5H) ppm.

Preparation C

Benzyl-6,6-d.ibrompenicillanat-la-oxide.

To a stirred solution of 13.4 g (0.03 mole) of benzyl 6,6-dibrompenicillanate in 200 ml of dichloromethane was added a solution of 6.12 g (0.03 mole) of 3-chloroperbenzoic acid in 100 ml of dichloromethane. ca. 0 ° C. Stirring was continued for 1.5 hours at ca. 0 ° C, after which the reaction mixture was filtered. The filtrate was washed successively with 5% sodium hydrogen carbonate solution and water and then dried (Na 2 SO 4). Removal of the solvent by evaporation in vacuo afforded 12.5 g of the title product as an oil. The oil was solidified by trituration under ether. By filtration, 10.5 g of benzyl 6,6-dibrompenicillanate 1-oxide was then obtained as a solid. The IR spectrum (CHCl3) showed absorptions at 1800 and 1750 cm -1. The NMR spectrum of the product (CDCl 3) showed absorptions at 1.3 (s, 3H), 1.5 (s, 3H), 4.5 (s, 1H), 5.18 (s, 2H), 5.2 (s). , 1H) and 7.3 (s, 5H) ppm.

46 DK 155740 B

Preparation D

4-Nitrobenzylpenicillanat.

Reaction of the triethylamine salt of penicillanic acid with 4-nitrobenzyl bromide by the method of Preparation B gave 4-nitrobenzylpenicillanate.

Preparation of 3-Phthalidylpenicilianate.

To a solution of 506 mg of penicillanic acid in 2 ml of N, N-dimethylformide was added 0/476 ml of diisopropylethylamine followed by 536 mg of 3-phthalidyl bromide. The mixture was stirred overnight, then diluted with ethyl acetate and water. The pH was adjusted to 3.0 and the layers separated. The organic layer was washed with water and then with pH 8.0 water, and then dried using anhydrous sodium sulfate. The dried ethyl acetate solution was evaporated in vacuo to give 713 mg of the title ester as an oil. The NMR spectrum (CDCl 3) showed absorptions at 1.62 (m, 6H), 3.3 (m, 2H), 4.52 (s, 1H), 5.23 (m, 1H) and 7.63 (m , 5H).

Preparation F

Pivaloyloxymethylpenicillanat.

To 3.588 g of 6,6-dibrompenicillanic acid in 10 ml of Ν, Ν-dimethylformamide was added 1.8 ml of diisopropylethylamine followed by 1.40 ml of chloromethyl pivalate. The mixture was stirred overnight, then diluted with ethyl acetate and water. The organic layer was separated and washed successively with water of pH 3.0 and water of pH 8.0. The ethyl acetate solution was dried (Na 2 SO 4) and then evaporated in vacuo to give pivaloyloxymethyl 6,6-dibrompenicillanate as an amber oil (3.1 g) which slowly crystallized.

The above ester was dissolved in 100 ml of methanol, then 3.1 g of 10% palladium-on-carbon and 1.31 g of potassium hydrogen carbonate were added in 20 ml of water. The mixture was shaken under hydrogen at atmospheric pressure until hydrogen uptake ceased. The reaction mixture was filtered and the methanol removed by evaporation in vacuo. The residue was partitioned between water and ethyl acetate at pH 8, and then the organic layer was separated. The latter was dried (Na 2 SO 4) and evaporated in vacuo to give 1.25 g of the title compound

Claims (7)

1-NL ^ //// in CT COOR X 6 wherein R has the same meaning as R or is a conventional penicillin-carboxy protecting group, after which removing the carboxy protecting group and, if desired, converting a obtained compound into a pharmaceutically acceptable salt by reaction with a base using conventional methods, or gb) to prepare compounds of formula I wherein R has the above meaning other than hydrogen, reacting a salt of penicillanic acid 1,1-dioxide with 3-phthalidyl chloride , 3-phthalidyl bromide, 4-crotonolacton-4-yl chloride, 4-crotonolacton-4-yl bromide, Y-butyrolacton-4-yl chloride, Y-butyrolacton-4-yl bromide or a compound of the formula f $ R1 0 I II 5 I II 5 QCOC-R1 XII or QC-0-C-0-R XIII 14 I4 RR 5 wherein R -R has the above meanings, and Q is chlorine or bromine, in a reaction inert solvent at a temperature in the range of 0 to 100 ° C. An analogous process for the preparation of penicillanic acid derivatives of the formula H, 0 "-s'0," 'CH3 Γ' '- (CH3) -' - \ 6 O 'COOR wherein R6 is hydrogen, 3-phthalidyl, 4-crotonolactonyl , Y-butyrolacton-4-yl or a group of formula R3 O R3 0 I II 5 I II 5 -COCR X or -COCOR XI I 4 I 4 RR wherein R and R are each hydrogen, methyl or ethyl and R is alkyl of 1-6 carbon atoms, and when R6 represents hydrogen, physiologically acceptable salts thereof, characterized in that A) oxidizes a compound of the formula OOH =, vCH ^ I 0'CH3 C · c ν '3' c 'V -S ν' V -.S ^ r-YY-CH3 II / rf> CH3 1_n_L J - nL CT ^ COOR1 CT ^ COOR1 \ .s - 3 or - "~ CH3 III ' Process according to claim 1a, characterized in that the oxidation is carried out using a metal permanganate or an organic peroxycarboxylic acid. 49 DK 155740 B Process according to claim 2, characterized in that the oxidation is carried out using potassium permanganate. Process according to claim 3, characterized in that the oxidation is carried out in a reaction-inert solvent at a temperature in the range of from approx. -20 to approx. 50 ° C. 4-Nitrobenzylpenicillanat. To a stirred solution of 2.14 g of penicillanic acid and 2.01 ml of ethyl diisopropylamine in 10 ml of Ν, Ν-dimethylformamide was added dropwise 2.36 g of 4-nitrobenzyl bromide at ca. 20 ° C. The mixture was stirred at room temperature overnight, then diluted with ethyl acetate and water. The layers were separated and the ethyl acetate layer was washed with water of pH 2.5 followed by water of pH 8.5. The ethyl acetate solution was then dried (1 SO 2) and evaporated in vacuo to give 3.36 g of the title compound. The NMR spectrum of the product (in CDCl 3) showed absorptions at 1.45 (s, 3H), 1.68 (s, 3H), 3.32 (m, 2H), 4.50 (s, 1H), 23 (m, 1H), 5.25 (s, 2H) and 7.85 (q, 4H) ppm. Process according to any one of claims 1-4, 16 characterized in that R and R are hydrogen. Process according to claim 1b), characterized in that R 1 is alkanoyloxymethyl having 3-8 carbon atoms. Process according to claim 6, characterized in that R is pivaloyloxymethyl.