GB2036027A - Novel penicillin compounds and the pharmaceutical application thereof - Google Patents

Abstract

Penicillanic acid derivatives of the general formula <IMAGE> wherein R<1> represents hydrogen or hydroxy, R<2> represents hydrogen or a saltforming cation or ester-forming group, and R<3> represents a group of the general formula <IMAGE> wherein R<4> represents hydrogen or optionally substituted lower alkyl, Q represents dimethylene, trimethylene, tetramethylene, optionally substituted o-phenylene or a divalent moiety derived from a 5 or 6 membered heterocyclic compound containing one or two nitrogen atom(s) wherein the two free bonds are situated adjacent to each other, and R<5> represents hydroxy, halogen, optionally substituted lower alkyl, optionally substituted aryl, optionally substituted benzyl, optionally substituted lower alkoxy, optionally substituted aryloxy, optionally substituted benzyloxy or a group of the general formula -NR<6>R<7>, wherein R<6> and R<7> are the same or different and each represents hydrogen, optionally substituted lower alkyl, cycloalkyl, aralkyl or optionally substituted aryl, or R<6> and R<7> together with the adjacent nitrogen atom form a 5 or 6 membered heterocyclic ring which may contain a further hetero atom selected from the group consisting of nitrogen, oxygen and sulphur, and enantiomers and, when more than one asymmetric carbon atom is present, also diastereoisomers and salts and, when R<2> represents a saltforming cation, hydrates thereof, are prepared by synthesis of either of the extracyclic amide linkages. The preferred derivative is alpha -(1-Ethyl-2- methoxy-2-oxo-1,3,2- diazaphosphorinano-3-carbonyl- amino)benzylpenicillin. There is also provided a pharmaceutical composition which comprises an effective amount of at least one derivative in accordance with the invention, in association with a suitable physiologically acceptable carrier or excipient, as well as the use of the derivatives of the invention as a therapeutic agent against a microorganism and as a growth promotor for ruminant animals.

Description

SPECIFICATION Novel penicillin compounds and the pharmaceutical application thereof.

The present invention relates to novel penicillins. The novel penicillins are therapeutically useful against a broad spectrum of microorganisms, especially against Proteus and Pseudomonas species. It is well-known that 6-(a-aminoacylamidp)penicillanic acid derivatives such as ampicillin, amoxicillin and epicillin are active against many gram positive and gram negative bacteria. These compounds do not show useful activity againstPseudomonas and indole positive Proteus species. 6-(a Carboxyacylamido)penicillanic acid derivatives such as carbenicillin have been developed and show rather good activity against Pseudomonas species but many strains of Pseudomonas and indole positive Proteus are rather insensitive to carbenicillin and similar penicillins.An object of the present invention is to provide penicillin derivatives which show useful activity against these microorganisms.

The present invention relates to novel therapeutically useful penicillanic acid derivatives of the general formula

wherein R1 represents hydrogen or hydroxy, R2 represents hyrogen or a salt forming cation or ester-forming group, and R3 represents a group of the general formula

wherein R4 represents hydrogen or optionally substituted lower alkyl, Q represents dimethylene, trimethylene, tetramethylene, optionally substituted o-phenylene or a divalent moiety derived from a 5 or 6 membered heterocyclic compound containing one or two nitrogen atom(s) wherein the two free bonds are situated adjacent to each other, and R5 represents hydroxy, halogen, optionally substituted lower alkyl, optionally substituted aryl, optionally substituted benzyl, optionally substituted lower alkoxy, optionally substituted aryloxy, optionally substituted benzyloxy or a group of the general formula -NR6R7, wherein R6 and R7 are the same or different and each represents hydrogen, optionally substituted lower alkyl, cycloalkyl, aralkyl or optionally substituted aryl, or R6 and R7 together with the adjacent nitrogen atom from a 5 or 6 membered heterocyclic ring which may contain a further hetero atom selected from the group consisting of nitrogen, oxygen and sulphur, and enantiomers and, when more than one asymmetric carbon atom is present, also diastereoisomers and salts and, when R2 represents a salt forming cation, hydrates thereof.

Throughout the present specification and claims, the term "lower alkyl", when used alone or in combination with other groups, designates a straight or branched alkyl group preferably containing at the most 6 carbon atoms, more preferred at the most 4 carbon atoms, e.g. methyl, ethyl, propyl, isopropyl and tert.butyl. The term "lower alkoxy" designates a straight or branched alkoxy group preferably containing at the most6 carbon atoms, more preferred at the most 4 carbon atoms, e.g.

methoxy, ethoxy, isopropoxy and tert.butoxy. As examples of salt-forming ions may be mentioned sodium, potassium, ammonium, calcium or substituted ammonium, e.g. tri(lower alkyl)amino, procaine or benzylamino. As examples of ester-forming groups may be mentioned groups of the general formula -CH20C0-R8 and -CHCH30C00-R8, wherein R8 represents lower alkyl which may be substituted by one or more of the substituents selected from the group consisting of lower alkoxy, lower alkyithio and halogen-lower alkyl, and as specific examples of such ester-forming groups may be mentioned acetoxymethyl, tert.butyl carbonyloxymethyl and 1 -(ethoxycarbonyloxy)ethyl. The above-mentioned saltforming cations and ester-forming groups are preferably pharmaceutically acceptable cations and groups.

It appears from the definitions that the lower alkyl, aryl, benzyl, alkoxy, aryloxy, benzyloxy and o- phenylene groups may be substituted, and in such cases there is preferably one substituent; however, said groups may carry more substituents, e.g. two or three substituents. As examples of such substituents may be mentioned halogen, e.g. chlorine or bromine, cyano, lower alkoxy, e.g. methoxy or ethoxy, thio-lower alkyl, trifluoromethylthio, lower alkoxycarbonyl, e.g. ethoxycarbonyl, carboxy, carboxamido, hydroxy, cycloalkyl and a group of the formula -NR6R7, wherein R6 and R7 each have the above-mentioned meaning.As examples of optionally substituted alkyl groups may be mentioned trihalogenoalkyl, e.g. 2,2,2-trichloroethyl and trichloromethyl, halogenoalkyl, e.g. 2-chloroethyl, aikoxyalkyl, e.g. methoxymethyl, cyanoalkyl, e.g. cyanomethyl and 2-cyanoethyl, trifluoromethylthioalkyl, e.g. trifluoromethylthiomethyl, alkoxycarbonylalkyl. e.g. ethoxycarbonylmethyl, and carboxyalkyl, e.g. a carboxymethyl.As examples of optionally substituted aryl groups may be mentioned o-, m- and p-tolyl, alkoxyphenyl, e.g. o-, m- and p-methoxyphenyl, mono- and dihalogenophenyl, e.g. o-, m- and p-chlorophenyl, 2,4-dichlorophenyl and 2,6-dichlorophenyl, mono- and dihydroxyphenyl, e.g. o-, m- and p-hydroxyphenyl and 2,4-dihydroxyphenyl, and hydroxyhalogenophenyl, e.g. 3-chloro-4hydroxyphenyl. As examples of optionally substituted benzyl groups may be mentioned alkoxybenzyl, e.g. o-, m- and p-methoxybenzyl, mono- and dihalogenobenzyl, e.g. o-, m- and p-chlorobenzyl and 2,4-dichlorobenzyl, mono- and dihydroxybenzyl, e.g. o-, m- and p-hydroxybenzyl and 2,4dihydroxybenzyl, and hydroxyhalogenobenzyl, e.g. 3-chloro-4-hydroxybenzyl.As examples of optionally substituted alkoxy groups may be mentioned ha loge noalkoxy, e.g. 2-chloroethoxy, trihalogenoalkoxy, e.g. 2,2,2-trichloroethoxy, cyanoalkoxy, e.g. 2-cyanoethoxy and cyanomethoxy, trifluoromethylthioalkoxy, e.g. trifluoromethylthiomethoxy, and N,N-di(lower alkoxy)aminoalkoxy, e.g. 2 (dimethylamino)ethoxy. As examples of optionally substituted aryloxy groups may be mentioned monoand dialkylphenoxy, e.g. o-, m- and p-tolyloxy, 2,4-dimethylphenoxy and 3,5-dimethylphenoxy, monoand dialkoxyphenoxy, e.g. o-, m- and p-methoxyphenoxy, 2,4-dimethoxyphenoxy and 3,4-dimethoxyphenoxy, and mono- and dihalogenophenoxy, e.g. o-, m- and p-chlorophenoxy, 2,4dichlorophenoxy and 2,6-dichlorophenoxy.As examples of optionally substituted benzyloxy groups may be mentioned mono- and dialkoxybenzyloxy, e.g. o-, m- and p-methoxybenzyloxy, 2,4dimethoxybenzyloxy and 3,4-dimethoxybenzyloxy, and mono- and dihalogenobenzyloxy, e.g. o-, m- and p-chlorobenzyloxy and 2,4-dichlorobenzyloxy. As examples of groups of the formula NReR7 may be mentioned amino, methylamino, isopropylamino, dimethylamino, diethylamino, halogenoalkylamino, e.g. 2-chloroethylamino, cyclohexylamino, anilino and benzylamino. As examples of groups of the formula -NR6R7, wherein R6 and R7 are moieties in a ring, may be mentioned piperidino, pyrrolo, pyrolidino and morpholino.

The lower alkyl group stated for the symbol R4 and the lower alkyl and aryl groups stated for the symbols R6 and R7 are preferably unsubstituted and, preferably, the diva lent moiety derived from a heterocyclic compound stated for the symbol Q is 6 membered.

Preferred compounds of the formula I are compounds wherein R3 represents a group wherein Q represents dimethylene or trimethylene, R4 represents ethyl, and R5 represents hydroxy, methyl, methoxy, ethoxy, 2,2,2-trich loroethoxy, phenoxy, benzyloxy and diethylamino, and enantiomers, diastereoisomers, salts and hydrates thereof, preferably physiologically acceptable salts thereof.

A further preferred subclass of compounds of the formula I is compounds of the general formula I, wherein R1 and R2 each have the above-mentioned meanings, R4 represents methyl, ethyl, propyl or isopropyl, R5 represents hydroxy or methoxy, and Q represents trimethylene. A preferred subclass of compounds within the last-mentioned subclass is compounds of the formula 1, wherein R1 and R2 each have the above-mentioned meanings, R4 represents ethyl, R5 represents hydroxy or methoxy, and Q represents trimethylene. Preferably R1 represents hydrogen, and R2 represents sodium, especially within the last-mentioned two preferred subclasses of compounds of the formula The compounds of formula I and salts and hydrates thereof may be prepared by the use of methods being analogously to methods used for the preparation of structurally similar penicillin derivatives. Thus, compounds of formula I or salts or hydrates thereof can, e.g., be prepared by a) reacting a compound of the general formula

wherein R9 represents hydrogen or a leaving group commonly connected to an amino group which is to be acylated, and R10 and R11, respectively, each have the same meaning as R1 and R2, respectively, or represents groups convertible thereto, or an acid addition salt or hydrate thereof with a compound of the general formula R12-R1 (IV) wherein R12 represents a leaving group commonly used as an activating group of an acylating agent, and R'3 has the same meaning as R3 or represents groups convertible thereto, or b) reacting a compound of the general formula

wherein R9 and R11 each have the above meanings, our a salt or hydrate thereof with a compound of the general formula

wherein R'O and R'3 each have the above meanings, or an activated derivative thereof, and, if necessary, subsequent to process variant a) or b) converting one or more of the groups R10, R" and R13, respectively, into the corresponding groups R', R2 and R3, respectively, and/or, if desired, converting one or more of the groups R', R2 and R3, respectively, into other groups within the definitions of R1, R2 and R3, respectively, and, if desired, converting a compound of formula I into a salt or into a hydrate thereof, or, if desired, converting a salt or a hydrate of a compound of the formula I into a compound of the formula I.

The term "group convertible thereto" designates, e.g., groups corresponding to the groups R' and R3, wherein any hydroxy or amino group contains protecting groups, and as examples of such protecting groups may be mentioned 2,2,2-trichloroethoxycarbonyl, benzyloxycarbonyl, benzyl, acetyl and a silicon or phosphorus atom carrying substituents selected from the group consisting at lower alkyl, halogenolower alkyl, aryl, aralkyl, lower alkoxy, halogeno-lower alkoxy, lower alkylthio, aralkoxy, di(lower alkyl)amino, lower alkoxyalkyl and lower alkylenedioxy, preferably tri(lower akyl)silyl, e.g. trimethylsilyl and tert.butyldimethylsilyl. The introdúction of said groups in the above compounds and the subsequent conversion of said groups into the desired groups are carried out by methods known per se.It is essential that the protecting groups can be removed easily without any undesired influence on the parent compound and, preferably, there are used protecting groups which can be removed by hydrolysis or hydrogenation. In case R11 represents a non pharmaceutically acceptable cation or group, it is preferably converted into a pharmaceutically acceptable cation or group by the use of methods known peruse. Preferably, R" represents an ester-forming group protecting the carboxy radical which can be easily removed after the reaction, e.g. by hydrolysis, hydrogenation or a substitution reaction using a basic or nucleophilic reagent, and which does not interfere with the reaction.As examples of easily removable protecting groups represented by R11 may be mentioned optionally substituted benzyl, e.g. pnitrobenzyl and p-methoxybenzyl, benzhydryl, phenacyl, e.g. p-halogeno-phenacyl, 2,2,2-trichloroethyl or radicals having a silicon or phosphorus atom carrying substituents selected from the group consisting of lower alkyl, halogeno-lower alkyl, aralkyl, lower alkoxy, lower alkylthio, alkoxy, di(lower alkyl)amino,r lower alkoxy-lower alkyl, lower alkylenedioxy and halogen The reaction ofa compound of formula Ill with a compound of formula IV according to process variant a) is in principle an acylation and, hence, it is obvious for those skilled in the art how the method is to be performed.The reaction is preferably carried out by adding a compound of the formula IV which may be dissolved in an organic solvent or a mixture of organic solvents such as an ether, e.g. diethyl ether, or acetone, to a compound of the formula III which may be suspended or dissolved in a solvent or a mixture of solvents such as a mixture of an ester, e.g. ethyl acetate, and water, or a salt or hydrate thereof. The reaction may be carried out at a temperature between -30 and +300C, preferably between -5 and +5 0C, e.g. by the use of an ice bath. The reaction time depends on the reaction temperature and may be in the range between 10 minutes and 10 hours, although it may even be shorter or longer. Preferably, the reaction mixture is stirred during the reaction.The yield may be increased by the addition of acid acceptors such as pyridine, triethylamine N,N-dimethylaniline, epoxides, e.g. 1 ,2-propylenoxide, or sodium hydrogencarbonate. Preferably, R9 represents hydrogen or trimethylsilyl, and R12 represents halogen such as chlorine, in the starting compounds. The compounds of formula I may be recovered from the reaction mixture by the use of methods known per se.

A general method for the preparation of a subclass of compounds of the formula IV, i.e.

compounds of the general formula

wherein R15 represents halogen, R16 represents halogen, a group convertible into hydroxy, optionally substituted alkyl, aryl, benzyl, lower alkoxy, aryloxy, benzyloxy or a group of the formula NReR7, wherein R6 and R7 each have the above meanings or represents groups convertible thereto, and R14 and Q1, respectively, each have the same meaning as R4 and Q, respectively, or represents groups convertible thereto, or salts thereof, are described in Example 6a and 6b or said compounds can be prepared analogously to the method described in said subexamples.The remaining compounds of the formula IV can be prepared by reacting a compound of the general formula

wherein R14, R16 and Q1 each have the above-mentioned meanings, with a compound of the general formula R18-COO-R17 (IX) wherein OR17 represents an activating group, e.g. p-nitrophenoxy, and R18 represents halogen, preferably chlorine.

The reaction of a compound of formula V with a compound of formula VI according to process variant b) is in principle an acylation and, hence, it is obvious for those skilled in the art how the method is to be performed. The reaction can be carried out in the presence of a condensing agent, e.g. a carbodiimide such as dicyclohexylcarbodiimide, optionally mixed with 1 -hydroxybenzotriazole. A compound of the formula VI may be activated by the methods known per se, e.g. by the conversion into acid halides, mixed anhydrides and activated esters. When using acid halides, R9 may be hydrogen, a substituted Si or P-atom, where Si may be substituted with lower alkyl, preferably trimethylsilyl, and where P may be substituted with dioxoalkylene, preferably dioxyethylene.When R9 is hydrogen, the reaction may be performed in the presence of an acid scavenger, e.g. organic or inorganic bases or epoxides. R1l represents a salt or a protecting group which can be removed after the acylation reaction.

Derivatives of compounds of the formula VI wherein the carboxylic group is protected, can be prepared analogously to the preparation of compounds of the formula I by the use of process variant a).

It is within the ken for those skilled in the art to judge which groups shourd be protected in the starting compounds when preparing compounds of formula I or the intermediates used for the preparation thereof.

Those starting materials for which the preparation is not described herein, are either known compounds or compounds which may be prepared in analogy with the preparation of known compounds or in analogy with known processes.

A compound prepared by any of the above methods may be isolated and purified in a manner known per se, e.g. by filtration, evaporation of the solvent and recrystallization.

The compounds of formula I according to the present invention show interesting pharmacological properties, in particular they are effective against a broad spectrum of microorganisms and especially active against Proteus and Pseudomonas species.

The compounds of formula I are preferably employed for therapeutic purposes in the form of nontoxic salts such as sodium, potassium, ammonium or calcium salts. As examples of other salts that may be used, may be mentioned non-toxic suitably crystallising salts with organic bases such as amines, for example trialkylamines, procaine and dibenzylamine.

In the treatment of bacterial infections, the antibacterial compounds of formula I according to the invention can be administered topically, orally and parenterally, in accordance with conventional procedures for antibiotic administration. They are administered in dosage units containing an effective amount of the active ingredient in combination with suitable physiologically acceptable carriers or excipients. The dosage units can be in the form of liquid preparations, such as solutions, suspensions, dispersions or emulsions or in a solid form, such as powders, tablets and capsules.

Accordingly, the invention includes pharmaceutical compositions comprising an effective amount of at least one compound of formula I, in association with a suitable physiologically acceptable carrier or excipient. Such pharmaceutical compositions can also include one or more therapeutic ingredients in addition to a compound of the formula I. The term "effective amount" as used herein in relation to the compounds of formula I designates an amount which is sufficient to destroy or inhibit the growth of susceptible microorganisms when administered in the usual manner, in other words an amount which is sufficient to control the growth of bacteria.The magnitude of an effective amount can be easily determined by those skilled in the art through standard procedures for determining the relative activity of antibacterial agents when utilized against susceptible organisms via the various available routes of administration. In this connection the effective amounts known for administering carbenicillin may be chosen as a starting point. A preferred tablet formulation for oral administration contains 250-1000 mg of a compound of the formula I per unit dosage which may be administered 1-4 times per day.

Suitable carriers and excipients may be any convenient physiologically acceptable ingredient which can serve to facilitate administration of the compound of formula I. Carriers may provide some ancillary function such as that of a diluent, flavour-masking agent, binding agent, action-delaying agent or stabilizer. As examples of carriers may be mentioned water, which may contain gelatin, acacia, alginate, dextran, polyvinylpyrrolidone, sodium carboxymethyi cellulose, aqueous ethanol, syrup, isotonic saline, isotonic glucose, starch, lactose or any other material commonly used in the pharmaceutical and veterinary industry.

The novel compounds of the formula I may also be used as growth promotors for ruminant animals such as cattle.

They are also useful in in vitro applications such as in liquids for disinfecting purposes at a concentration of, e.g. 0.1-1% by weight, dissolved or suspended in a suitable inert carrier for application by washing or spraying.

The process for the preparation of the above-mentioned novel compounds is illustrated by the following examples which, however, are not to be construed as limiting. The examples especially illustrate the preferred embodiments of the process. However, the yields obtained can in some cases be increased substantially.

EXAMPLE 1 a) 1 -Ethyl-2-methoxy-2-oxo-1 ,3,2-diazaphosphorinane.

A solution of 1.5 g of methyl phosphorodichoroidate in 3 ml of diethyl ether was added to a mixture of 1.02 g of 3-(ethylamino)propylamin, 3.36 g of DABCO (designates 1,4-diazabicyclo (2,2,2)octan) and 60 ml of diethyl ether in an ice bath. The mixture was stirred for 3 hours at room temperature. The precipitate formed was filtered off and the filtrate was concentrated to dryness to give an oil, which was subjected to silicagel column chromatography. The eluate of chloroform was evaporated to give 1 -ethyl-2-methoxy-2-oxo-1 ,3,2-diazaphosphorinane.

NMR(CDCI3)8: 1.10(3H, t, CH2CH3, J = 6.0 Hz), 1.50-1.90 (2H, m, CH2CH2CH2), 2.70-3.50 (6H, m, CH2 x 3) and 3.60 (3H, d, POCH3, J = 17.0 Hz).

b) 1 -Ethyl-2-methoxy-2-oxo- 1 ,3,2-diaza phosphorinano-3-carbo nyl chloride.

A solution of 220 mg of trimethylchiorosilane in 2 ml of methylene chloride was added to a mixture of 300 mg of 1-ethyl-2-methoxy-2-oxo-1 ,3,2-diazaphosphorinane. 400 mg of triethylamine and 5 ml of methylene chloride in an ice bath. The mixture was stirred for 30 minutes in the same bath and added to a solution of about 250 mg of carbonyl chloride in 2.5 ml of methylene chloride in a dryice/methanol bath. The temperature of the mixture was gradually risen to room temperature during 1 hour. After evaporation of the solvent, diethyl ether was added to the residue to form a precipitate which was filtered off. The filtrate was concentrated to give 1 -ethyl-2-methoxy-2-oxo-1 ,3,2- diazaphosphorinano-3-carbonyl chloride as a colourless oil in a quantitative yield.

IR D (max., CHCI3) cm 1: 1730(C=0).

c) a-( 1 -Ethyl-2-methoxy-2-oxo- 1,3,2-d iazaphosphorina no-3-carbonylami no) benzy Ipenicillin.

A solution of 3.5 g of 1 -ethyl-2-methoxy-2-oxo- 1 ,3,2-diazaphosphorinano-3-carbonyl chloride in 5 ml of diethyl ether was added to a suspension of 5.8 g of ampicillin trihydrate, 2.5 g of sodium bicarbonate, 40 ml of water and 20 ml of ethyl acetate in an ice bath. The mixture was stirred for 6 hours in the bath. The ethyl acetate layer was separated off and thereafter the aqueous layer was washed with ethyl acetate, acidified with 2 N HCI and thereafter extracted with ethyl acetate. The extract was washed three times with ice cold 2 N HCI and with water, dried and thereafter evaporated to leave an oil, which was dissolved in methylene chloride and thereafter evaporated. This treatment was repeated three times to remove a small amount of the ethyl acetate from the oil.After the treatment, 4.8 g (yield: 77%) of a-(1-ethyl-2-methoxy-2-oxo-1 ,3,2-diazaphosphorinano-3- carbonylamino)benzylpenicillin was obtained as a white powder.

IR P (max., KBr) cm-': 3300 (NH), 1785 (p-lactam C = O),1740 (COOH), 1660 and 1680 (CONH).

NMR (CDCI3) : 1.00--1.30 (3H, m, CH2CH3), 1.60-1.90 (2H, m, CH2CH2CH2), 2.80-3.40 (6H, m, CH2 x 3), 3.55 and 3.75 (3H, each d, POCH3, J = 10.0 Hz), 4.35 (1 H, s, C3-H), 5.25-5.80 (3H, m, C5 and C6-H and PhCH), 6.70-6.95 (1 H, m, NH), 7.50 (1 H, m, NH) and 8.80 H, m, COOH). "Ph" represents phenyl.

EXAMPLE 2 a) 1 -Ethyl-2-ethoxy-2-oxo- 1,3,2-diazaphosphorinane.

A solution of 489 mg of ethyl phosphorodichloridate in 1 ml of diethyl ether was added to a solution of 306 mg of 3-(ethyiamino)propylamine and 1 g of DABCO in 20 ml of diethyl ether at room temperature. The mixture was stirred for 3 hours at room temperature. DABCO hydrochloride formed was filtered off and the filtrate was concentrated in vacuo at 800C to give 400 mg (yield: 69%) of 1 ethyl-2-ethoxy-2-oxo-1 ,3,2-diazaphosphorinane as a colourless oil.

NMR (CDCI3) 8: 1.20 and 1.40 (6H, each t, CH3 x 2, J = 8.0), 1.50--2.00 (2H, m, Ch2CH2CH2), 2.84 (1 H, s, NH, disappeared with D20), 2.90-3.30 (6H, m, CH2NCH2CH2CH2N), 3.99 (2H, quintet, CH2, J = 6.0 Hz), MS m/e: 1 94 (M+) and 179 (M±1 5, base peak).

b) 1 -Ethyl-2-ethoxy-2-oxo-1 ,3,2-diazaphosphorinano-3-carbonyl chloride.

A solution of 652 mg of trimethylchlorosilane in 2 ml of methylene chloride was added to a mixture of 776 mg of 1 -ethyl-2-ethoxy-2-oxo-1 ,3,2-diazaphosphorinane, 608 mg of triethylamine and 8 ml of methylene chloride in an ice bath. The mixture was stirred for 30 minutes at the same temperature. After stirring, the reaction mixture was cooled in a dry-ice methanol bath and 600 mg of diphosgene in 2 ml of methylene chloride was added to the reaction mixture over a period of 1 minute.

The temperature of the solution was raised gradually to room temperature over a period of 1 hour. After the reaction, the precipitate formed was filtered off and the filtrate was concentrated to give 1 -ethyl-2 ethoxy-2-oxo- 1 3,2-diaza phosphorinano-3-carbonyl chloride in a quantitative yield.

IRv(max., CHCI3) cm~1: 1720 (COCI). MS m/e: 258 (M+ + 2), 256 (M+) and 241 (M±1 5, base peak).

c) a-( 1 -Ethyl-2-ethoxy-2-oxo- 1 ,3,2-diazaph osphorinano-3-carbonyla mino)benzylpenicillin .

A solution of 256 mg of 1 -ethyl-2-ethoxy-2-oxo 1 ,3,2-diazaphosphorinano-3-carbonyl chloride in 0.5 ml of diethyl ether was added to a mixture of 403 mg of ampicillin trihydrate, 1 70 mg of sodium bicarbonate, 4 ml of water and 2 ml of ethyl acetate in an ice bath. The mixture was stirred for 1 hour at the same temperature. After the reaction the ethyl acetate layer was separated off and the aqueous layer was washed with ethyl acetate and separated off. Ethyl acetate was added to the aqueous solution, which was acidified with 1 N HCI and thereafter stirred for 30 minutes at OOC. The ethyl acetate layer was separated, washed with ice cold 1 N HCI and water and thereafter dried.The solvent was evaporated off to give an oily residue containing a-( 1 -ethyl-2-ethoxy-2-oxo-1 ,3,2- diazaphosphorinano-3-carbonylamino)benzylpenicillin, which was converted into the corresponding potassium salt using ethyl hexanoate potassium salt to give 200 mg (yield: 55%) of the potassium salt as a white powder.

IR V (max., KBr) cm-': 1765 (,B-lactam C =0), 1660 (amide C = 0) and 1600 (COO-).

NMR (DMS0-d6) : 1.00-1.80 (8H, m, CH3 x 2 and CH2CH3 x 2 and CH2CH2CH2), 1.40 and 1.52 (6H, m, each s,C2-CH3), 2.80-3.20 (4H, m, CH2CH2CH2), 3.84(1 H, s, C3-H), 3.0-4.4 (2H, m, CH2), 5.20--5A0 (2H, m, C5-H and Ph-CHCO), 5.60 (1 H, q, C6-H, J = 4.0 and 6.0), 7.20-7.50 (5H, m, aromatic protons), 8.60-8.78 (1H, m, NH) and 8.82-9.00 (1H, m NH).

EXAMPLE 3 a-( 1 -Ethyl-2-ethoxy-2-oxo- 1 3 ,2-diazaphosphorinano-3-carbonylamino)-p- hydroxybenzylpenicillin.

A solution of 421 mg of 1-ethyl-2-ethoxy-2-oxo-1,3,2-diazaphosphorinano-3-carbonyl chloride in 1 ml of diethyl ether was added to a suspension of 256 mg of amoxicillin, 170 mg of sodium bicarbonate in 4 ml of water and 2 ml of ethyl acetate in an ice bath. The mixture was stirred for 1 hour in an ice bath. The ethyl acetate was separated off to give an aqueous layer which was washed with ethyl acetate, acidified with 1 N HCI, and thereafter extracted with ethyl acetate. The extract was washed with ice-cold 1 N HCI and with water, dried with sodium sulphate and thereafter evaporated to give a residue. The free acid was obtained as a white powder in a yield of 52%.

IR V (max., KBr) cm-' (as potassium salt): 1 765 (p-lactam C = 0), 1660 (amide C = 0) and 1600 (COO~).

NMR (CDCl3 + DMSO-d6) : 1.15 (6H, t, CH2CH3 x 2, J = 6.0 Hz), 1.48 and 1.57 (6H, each s,CH3 x 2), 2.80-3.30 (4H, m NCH2C H2CH2), 3.40 (2H, q. NCH2CH3,J = 6.0 Hz), 3.60-4.20 (2H, m CH3CH20), 4.20 (1 H, s, C3-H, 5.30-5.60 (3H, m, Ce and Ce-Hand PhCH), 6.70 and 7.18 (4H, each d, aromatic protons, J = 8.0 Hz) and 8.10-8.70 (2H, m, NH x 2).

EXAMPLE 4 a)l -Ethyl-2-(2,2,2-trichloroethoxy)-2-oxo-l ,3,2-diazaphosphorinane.

A solution of 800 mg of trichloroethyl phosphorodichloridate (vide W. Gerrarol, W. J. Green and R.

J. Phillips: J. Chem Soc., 1954, 1 148) in 3 ml of diethyl ether was added to a mixture of 3.10 mg of 3 (ethylamino)propylamine, 1 g of DABCO and 20 ml of diethyl ether in an ice bath. The mixture was stirred for 3 hours at room temperature. The precipitate formed was filtered off and the filtrate was concentrated in vacuo at 70-800C to leave 860 mg (yield: 86%) of 1 -ethyl-2-(2,2,2-trichloroethoxy)- 2-oxo-1 3,2-diazaphosphorinane as a colourless oil.

NMR(CDCl3): 1.10(1H,t,CH3, J = 10.0 Hz), 1.50--2.00 (2H, m, CH2CH2CH2),2.90-3.50(6H,m, CH2 x 3) and 4.42 (2H, d, CH2OP, J = 5.0 Hz).

b) 1 -Ethyl-2-(2,2,2-trichloroethoxy)-2-oxo-1 ,3,2-diazaphosphorinano-3-carbonyl chloride.

A solution of 120 mg of trimethylchlorosilane in 1 ml of methylene chloride was added to a mixture of 295 mg of 1 -ethyl-2-(2,2,2-trichloroethoxy)-2-oxo-1 ,3,2-diazaphosphorinane, 3 ml of methylene chloride and 120 mg of triethylamine and refluxed for 1 hour. The mixture was added to a 10% carbonyl chloride solution in 2 ml of methylene chloride in a dry ice/aceton bath and stirred for 2 hours at room temperature. The solvent was removed to give a residue which was extracted with diethyl ether. The evaporation of the solvent afforded 310 mg of 1 -ethyl-2-(2,2,2-trichloroethoxy)-2-oxo-1 ,3,2- diazaphosphorinano-3-carbonyl chloride as a colourless oil.

IR V (max., CHCI3) cm-l: 1730 (C = 0).

c) a-( 1 -Ethyl-2-(2,2,2-trichloroethoxy)-2-oxo-1 ,3,2-diazaphosphorinano-3- carbonylamino)benzylpenicillin.

A solution of 310 mg of 1 -ethyl-2-(2,2,2-trichloroethoxy)-2-oxo-1 ,3,2-diazaphosphorinano-3- carbonyl chloride in 3 ml of diethyl ether was added to a mixture of 145 mg of sodium bicarbonate, 347 mg of ampicillin trihydrate, 6 ml of water and 3 ml of ethyl acetate in an ice bath and the mixture was stirred for 6 hours in the same bath. The aqueous solution was washed well with ethyl acetate, acidified with 2 N HCI, and thereafter extracted with ethyl acetate. The extract was washed with 2 N HCI and with water and dried and thereafter evaporated to give a residual oil, to which methylene chloride was added and the mixture was evaporated in order to remove a small amount of ethyl acetate to give 1 70 mg of c-( 1 -ethyl-2-(2,2 ,2-trich loroethoxy)-2-oxo-1 ,3,2-d iazaphosphorinano-3carbonylamino)benzylpenicillin as a powder.

IR V (max., KBr) cm-1: 3300 (NH), 1 785 (p-lactam C = O), 1745 (COOH) and 1 680 (CONH).

NMR (CDCI3) b: 1.00--1.30 (3H, m CH3), 1.45 and 1.48 (6H, each s, CH3 x 2), 1.60-2.00 (2H, m, CH2CH2CH2), 2.80-3.50 (6H, m, CH2 x 3), 4.30 (1 H, s, C3-H), 4.25-4.65 (2H, m, CCI3CH2), 5.43 (1 H, s, PhCH), 5.30-5.75 (2H, m, C5 and C,--H), 6.607.00 ( 1 H, m, NH), 7.27 (5H, s, aromatic protons), 8.50-8.85 (1H, m, NH) and 9.50 (1H, broad s, COOH).

EXAMPLE 5 a)l -Ethyl-2-phenoxy-2-oxo- 1 ,3,2-diazaphosphorinane.

A solution of 930 mg of phenyl phosphorodichloroidate in 10 ml of diethyl ether was added to a mixture of 450 mg of 3-(ethylamino)propylamine, 980 mg of DABCO and 50 ml of diethyl ether and the mixture was stirred for 3 hours at room temperature. The resulting precipitate was removed from the reactant by filtration. The filtrate was evaporated to give 800 mg of a colourless oil, which was chromatographed on 1 6 g of silica gel. The eluate of chloroform containing 3% of methanol was evaporated to give 700 mg (yield: 66%) of 1 -ethyl-2-phenoxy-2-oxo-1 ,3,2-diazaphosphorinane as an oil.

NMR (CDCI3) : 1.1 (3H, t, CH2CH3, J = 8.0 Hz), 1.552.00 (2H, m, CH2CH2CH2), 2.70-3.50 (7H, m, CH2 x 3 and NH) and 7.20 (5H, s, aromatic protons).

b) 1 -Ethyl-2-phenoxy-2-oxo-1 ,3,2-diaza phosphorina no-3-ca rbonyl chloride.

A solution of 400 mg of trimethylchlorosilane in 4 ml of methylene chloride was added to a mixture of 100 mg of 1 -ethyl-2-phenoxy-2-oxo-1 ,3,2-diazaphosphorinan, 370 mg of triethylamine and 10 ml of methylene chloride in an ice bath and the mixture was stirred for 30 minutes on the bath. The mixture was added to a 10% carbonyl chloride solution in. 5 ml of methylene chloride in a dry ice/ethanol bath and stirred for 1 hour on the bath. The solvent was evaporated off to give a residue which was extracted with diethyl ether. The solvent was removed to give 800 mg 1 -ethyl-2-phenoxy-2oxo-1 ,3,2-diazaphosphorinano-3-carbonyl chloride as a colourless oil.

IR V (max., CHCl3) cm-': 1630 (C = 0).

c) a-( 1 -Ethyl-2-phenoxy-2-oxo-1 ,3,2-diazaphosphorinano-3-carbonylamino)benzylpenicillin.

A mixture of 600 mg of 1 -ethyl-2-phenoxy-2-oxo-1 ,3,2-diazaphosphorinano-3-carbonyl chloride and 6 ml of diethyl ether was added to a suspension of 800 mg ampicillin trihydrate, 340 mg of sodium bicarbonate, 10 ml of water and 5 ml of ethyl acetate in an ice bath and stirred for 5 hours on the same bath. The mixture was washed with ethyl acetate. The aqueous layer was acidified with 2 N HCI, extracted with ethyl acetate and washed with 2 N HCI and with water. After drying, the solvent was removed to give an oil, to which methylene chloride was added and the mixture was evaporated in vacuo to remove a small amount of ethyl acetate giving a-(1-ethyl-2-phenoxy-2-oxo-1 3,2- diazaphosphorinano-3-carbonylamino)benzylpenicillin as a powder.

IR V (max., KBr) cm-': 3300 (NH),1785 ( -lactam C = O),1745 (COOH) and 1 680 (CONH).

NMR (CDCI3) 6: 1.12 (3H, t, CH3, J = 5.5 Hz), 1.35-1.55 (6H, m, CH3 x 2), 1.60-2.00 (2H, m, CH2CH2CH2), 2.70-3.50 (6H, m, CH2 x 3), 4.25 and 4.30 (1 H, each s, C3-H), 5.1 5-5.50 (3H, m, C5 and C5-H and PhCH) and 7.05-7.35 (1 OH, m, aromatic protons).

EXAMPLE 6 The following process can be used for the preparation of compounds of the general formula

wherein R20 represents alkyl, aryl or benzyl.

a) A mixture of 7 mmoles of 2-(ethylamino)ethylamine, 14 mmoles of triethylamine and 5 ml of diethyl ether was added to a solution of 7 mmoles of a phosphorodichloridate of the general formula R200(P = O)Cl2 (XXI) wherein R20 has the above-mentioned meaning, in 20 ml of diethyl ether in an ice bath and the mixture was stirred for 5 hours at room temperature. The precipitate were filtered off and the filtrate was evaporated to give a phosholane of the general formula

wherein R20 has the above-mentioned meaning.

b) A solution of 5 mmoles of trimethylchlorosilane in 5 ml of methylene chloride was added to a mixture of 4.57 mmoles of a phospholane of the general formula XXII, 5 mmoles of triethylamine and 7 ml of methylene chloride in an ice bath and the mixture was stirred for 1 hour in the same bath. The mixture was added to a 10% carbonyl chloride solution in 9 ml of methylene chloride in a dry ice/ethanol bath and the mixture was stirred for 1 hour in the bath and the stirring was continued for additionally 1 hour at room temperature. The solvent was removed to give a residue which was extracted with diethyl ether. The diethyl ether was evaporated to give a phospholane carbonyl chloride of the general formula

wherein R20 has the above-mentioned meaning.

c) A solution of 4.2 mmoles of a phospholane carbonyl chloride of the general formula XXIII in 5 ml of diethyl ether was added to a suspension of 4.2 mmoles of ampicillin trihydrate, 8A mmoles of sodium bicarbonate, 1 5 ml of water and 7 ml of ethyl acetate in an ice bath and the mixture was stirred for 5 hours in the bath. The ethyl acetate was separated off and the aqueous solution was acidified with 2 N HCI and extracted with ethyl acetate. The ethyl acetate layer was washed with 2 N HCI and with water and thereafter dried. The solvent was evaporated to give a compound of the general formula XX as a powder.

EXAMPLE 7 al -Ethyl-2-benzyloxy-2-oxo- 1 ,3,2-diazaphosholane.

A solution of 1.08 g of benzylalcohol, 1.01 g of triethylamine and 5 ml of diethyl ether was added to 1.53 g of phosphorus oxychloride in 30 ml of diethyl ether and the mixture was stirred for 30 minutes on a dry ice/acetone bath. To the mixture a mixture of 0.88 g of 2-(ethylamino)ethylamine, 4.04 g of triethylamine and 5 ml of diethyl ether was added, the mixture was stirred for 1 hour on the bath, and thereafter the stirring was continued for 1 hour at room temperature. The precipitate formed were filtered off and the filtrate was concentrated to give 1.39 g (yield: 58%) of 1 -ethyl-2-benzyloxy-2oxo-1 ,3,2-diazaphospholane as a colourless oil.

NMR (CDCl3) #: 1.10 (3H, t, CH J = 6.0 Hz), 2.65-3.50 (6H, m, CH2 x 3), 4.93 (2H, d, POCH2 J = 9.0 Hz) and 7.30 (5H, s, aromatic protons).

b) 1 -Ethyl-2-benzyloxy-"2-oxo-1 ,3 ,2-diazaphospholano-3-ca rbonyl chloride.

The compound was prepared analogously to the process described in Example 6b and was obtained as a colourless oil in a yield of 75%.

IR P (max., CHCl3) cm-': 1735 (C=O).

c) &alpha;-(1 -Ethyl-2-benzyloxy-2-oxo-1 ,3 ,2-diaza phospholano-3-ca rbonylamino)benzylpenicil lin.

The compound was prepared analogously to the process described in Example 6c and was obtained as a white powder in a yield of 60%.

IR v (max., KBr) cm-1 1780 (ss-lactam C = O), 1735 (COOH) and 1660 CONH).

NMR (CDCl3) #: 1.13 (3H, t, CH3, J = 7.5 Hz), 1.58 (6H, s, CH3 x 2), 2.80-3.90 (6H, m, CH2 x 3), 4.35 (1 H, ,s,C3-H), 4.85-5.80 (5H, m, CH2OP, C5 and CeH, and PhCH), 6.60-7.00 ( 1 H, m, NH), 7.25-7.60 ( 1 OH, m, aromatic protons) and 7.95 (2H (2H, broad s, NH and COOH).

EXAMPLE 8 a) 1-Ethyl-2-benzyloxy-2-oxo-1,3,2-diazaphosphorinane.

The compound was prepared analogously to the process described in Example 7a and was obtained as a colourless oil in a yield of 98%.

NMR (CDCl3) S: 1.10 (3H, t, CH3, J = 6.0 Hz), 1.40--1.80 (2H, m, CH2CH2CH2), 2.75-3.50 (6H, m, CH2 x 6), 4.95 (2H, d, POCH2, J = 8.5 Hz) and 7.30 (5H, s, aromatic protons).

b) 1 -Ethyl-2-benzyloxy-2-oxo-1 ,3,2-diazaphosphorinano-3-carbonyl chloride.

The compound was prepared analogously to the process described in Example 6b and was obtained as a colourless oil in a yield of 89%.

IR V (max., CHCI3) cm-1: 1730 (C = 0).

c) &alpha;-(1 -Ethyl-2-benzyloxy-2-oxo-1 ,3,2diazaphosphorinano-3-carbonylamino)benzylpenicillin.

The compound was prepared analogously to the process described in Example 6c and was obtained as a white powder in a yield of 60%.

IR v (max., kBr) cm-1: 1 i85 (ss-lactam C = 0); 1 735 (COOH) and 1650-1690 (CONH).

NMR (CDCI3) S: 1.00-2.00(11 H, m, CH3 x 3 and CH2CH2CH2), 2.70-3.50 (6H, m, CH2 x 3), 4.28 and 4.32 (1H, a pair of s, C3- H), 4.70--5.15 (2H, m, CH20P), 5.30-5.70 (3H, m, PhCH, and C5 and C5-K) and 7.20--7.50(10H, m, aromatic protons).

EXAMPLE 9 a) 1-Ethyi-2-diethylamino-2-oxo-1,3,2-diazaphosphorinane.

The compound was prepared analogously to the process described in Example 6a and was obtained as a colourless oil in a yield of 99%.

NMR (CDCI3) S: 1 .00--2.30 (9H, m, CH3 x 3), 2.60-2.95 (2H, m, CH2CH2CH2 and 3.70-4.55 (1 OH, m, CH2 x 5).

b) 1 -Ethyl-2-diethylamino-2-oxo-1 ,3,2-diazaphosphorinano-3-ca rbonyl chloride.

The compound was prepared analogously to the process described in Example 6b. Although the crude product showed many spots on thin layer chromatography it was subjected to the following reaction without purification.

IR v (max., CHCI3) cm-': 1730 (COCI).

c) a-( 1 -Ethyl-2-diethylamino-2-oxo-1 ,3,2diazaphosphorinano-3-carbonylamino)benzylpenicillin.

The compound was prepared analogously to the process described in Example 6c and was obtained as a white powder.

IR V (max., KBr) cm-1: 1780 (,B-lactam C = O), 1730 (COOH) and 1 670 (CONH).

NMR (CDCI,) S: 0.80--1.80(17H, m, CH3 x 5 and CH2CH2CH2), 2.703.50 ( 1 OH, m, CH2 x 5), 4.30-4.32 ( 1 H, a pair of s, C3-H), 5.25-5.70 (3H, m, PhCH, and C5 and C5-H) and 7.32 (5H, s, aromatic protons).

EXAMPLE 10 a) 1 -Ethyl-2-methyl-2-oxo-1 ,3,2-diazaphosphorinane.

To a solution of 0.70 g (6.9 mmoles) of 3-(ethylamino)propylamine and 1.41 g (14 mmoles) of triethylamine in 10 ml of methylene choride was added 1.0 g (7.5 mmoles) of methanephosphonic acid dichloride in 10 ml of methylene chloride dropwise with cooling in an ice bath. After stirring for 16 hours, the precipitate was filtered off and the filtrate was evaporated. To the residue was added ether and the precipitate was filtered off. The filtrate was evaporated to give 0.40 g (yield: 36%) of 1 -ethyl-2 methyl-2-oxo-1 ,3,2-diazaphosphorinane as a colourless oil.

NMR (CDCl3) S: 1.10 (3H, t, CH3CH2), 1.40 (3H, d, CH3-P, J = 15.0 Hz), 1.50-2.00 (2H, m, CH2CH2CH2) and 2.60-3.50 (7H, m, NCH2 x 3, NH).

b) &alpha;-(1-Ethyl-2-methyl-2-oxo-1,3,2-diazaphosphonnano-3-carbonylamino)benzylpenicillin.

To a solution of 200 mg of 1 -ethyl-2-methyl-2-oxo-1 ,3,2-diazaphosphorinane and 293 mg of triethylamine in 4 ml of methylene chloride was added 161 mg of trimethylchlorosilane in 1.5 ml of methylene chloride and the mixture was thereafter refluxed for 1 hour. To 183 mg of carbonyl chloride in 1.83 ml of methylene chloride was added the previously prepared solution dropwise with cooling in a dry-ice/methanol bath. The mixture was stirred for 2 hours and 30 minutes. During this time, the temperature of the mixture was gradually risen to room temperature. The solvent was evaporated, dioxane was added to the residue and the mixture was filtered.The filtrate was concentrated to give 240 mg of a pale yellow oil. 1 50 mg of the oil in 0.2 ml of dioxane was added to a suspension of 270 mg of ampicillin trihydrate, 113 mg of sodium bicarbonate, 3 ml of water and 1.5 ml of ethyl acetate in an ice bath. The mixture was stirred for 6 hours in the bath. The ethyl acetate layer was separated off and thereafter the aqueous layer was washed with ethyl acetate. The extract was washed with 2 N HCI and thereafter with water, dried and thereafter evaporated to leave an oil which was triturated with ether to give &alpha;-( -ethyl-2-methyl-2-oxo-1,3,2-diazaphosphorinano-3-carbonylamino)benzylpenicillin as a white powder.

IR V (max., KBr) cm-1: 1785 (,B-lactam C = O),1735 (COOH) and 1690--1660 (CONH).

NMR (CDCI3) S: 1.00--1.35 (3H, m, CH2CH2), 1.43-1.58

1.55 and 1.73 (3H, each d, p-CH3, J = 1 5.0 Hz), 1.50-2.00 (2H, m (CH2), 2.80-3.40 (6H, m, N-CH2 x 3), 4.34 (1 H, s, C3-H), 5.30-5.80 (3H, m, C5 and CeH and PhCH), 6.8-7.1 H, m, NH), 7.38 (5H, s, CeHe) and 8.45-8.80 (1 H, m, NH).

EXAMPLES 11-15 The compounds stated in Examples 11-15 were prepared analogously to the process described in Examples 6a-6c.

EXAMPLE 11 a) 1 -Ethyl-2-methoxy-2-oxo- 1 ,3,2-d iazaphospholane.

Colourless oil; yield: 67%.

NMR (CDCl3) S: 1.20 (3H, t, CH2CH3, J = 7.5 Hz), 2.80--3.60 (7H, m, CH2 x 3 and NH) and 3.65 (3H, d, CH3OP,J= 13.5Hz).

b) 1 -Ethyl-2-methoxy-2-oxo- 1,3 2-diaza phosp holano-3-carbonyl chloride.

Colourless oil; yield: 93%.

IR V (max., CHCl3) cm-1: 1735 (COCI).

c) a (1 -Ethyl-2-methoxy-2-oxo- 1 ,3,2-diazaphospholano-3-carbonyla mino)benzylpenicillin.

White powder; yield: 27%.

IR u (max., KBr) cm-': 3350 (NH), 1785 (ss-lactam C C = O) and 1740 (COOH) and 1670 (CONH).

NMR (CDC3) #: 1.15 (3H,t,CH2CH3, J = 7.0 Hz), 1.48 (6H, s, CH3 x 2), 2.65-3.50 (6H, m, CH2 x 3), 3.55 and 3.76 (3H, double d, CH3OP, J = 1 2.0 Hz), 4.30 (1 H, s, C3-H), 5.30-5.75 (3H, m, C5 and CeH and PhCH), 6.80 ( 1 H, d, NH, J = 7.5 Hz) and 7.33 (5H, s, aromatic protons).

EXAMPLE 12 a) 1 -Ethyl-2-ethoxy-2-oxo-1 ,3,2-diaza phosphola ne.

Colourless oil; yield: 58%.

NMR (CDCI3) S: 1.08-1.50 (6H, m, CH2CH3), 2.85-3.50 (7H, m, CH2 x 3 and NH) and 3.70-4.1 5 (2H, m, CH2OP).

b) 1 -Ethyl-2-ethoxy-2-oxo- 1,3,2-diaza phospholano-3-carbonyl chloride.

Colourless oil; yield: 95%.

IRv(max.,CHCl3) cm-1: 1730(COCI).

c) &alpha;-(1 -Ethyl-2-ethoxy-2-oxo-1 ,3,2-diazaphosholano-3-carbonylamino)benzylpenicillin.

White powder; yield: 46%.

IR V (max., KBr) cm-1: 3350 (NH), 1785 (p-lactam C = O), 1735 (COOH) and 1665 (CONH).

NMR (CDCl3) S: 1.05-1.40 (6H, m, CH2CH3 x 2), 1.45 and 1.48 (6H, m, NCH2 x 3), 3.83--4.25 (2H, m, OCH2), 4.34(1 H, s, C3-H), 5.35-5.80 (3H, m, C5 and C5-H and PhCH), 6.85 (1H, m, NH), 7.35 (5H, s, aromatic protons) and 7.92 (2H, s, NH and COOH).

EXAMPLE 13 a) 1 -Ethyl-2-(2,2,2-trichloroethoxy)-2-oxo-1 3,2-diazaphospholane.

Colourless oil; yield: 97%.

NMR (CDCl3) S: 1.20 (3H, t, CH2CH3), J = 7.0 Hz), 2.80-3.50 (6H, m, CH2 x 3), 4.20-4.50 (1 H, m, NH) and 4.50 (2H, d, CH2OP, J = 7.7 Hz).

b) 1-Ethyl-2-(2,2,2-trichloroethoxy)-2-oxo-1,3,2-diazaphospholano-3-carbonyl chloride.

Colourless oil; yield: 97%.

IRw(max., CHCl3) cm-1: 1740 (COCI).

c) -(1 -Ethyl-2-(2 ,2 -Ethyl-2-(2,2,2-trichloroethoxy)-2-oxo-1 ,3,2-diazaphospholano-3- carbonylamino)benzylpenicillin.

White powder; yield: 49%.

IR V (max., KBr) cm-1: 3350 (NH), 1785 (ss (ss-lactam C=O), 1735 (COOH) and 1665 (CONH).

NMR (CDCI3) S: 1.10-1.40 (3H, m, CH2CH3), 1.47 (6H, s, CH3 x 2) 2.80-3.90 (6H m, CH3 x 3), 4.33 ( 1 H, s, C3-H), 4.30--4.80 (2H, m, CH2OP), 5.30-5.80 (3H, m, C5 and C6-H and PhCH), 7.35 (5H, s, aromatic protons) and 8.65 (2H, broad s, NH and COOH).

EXAMPLE 14 a) 1-Ethyl-2-phenoxy-2-oxo-1,3,2-diazaphospholane.

Colourless oil; yield: 88%.

NMR (CDCl3) S: 1.08 (3H, t, CH2CH), J = 7.0 Hz), 2.70---3.45 (6H, m, CH2 x 2), 3.60-4.00 (1 H, m, NH) and 7.22 (5H, s, aromatic protons).

b) 1-Ethyl-2-phenoxy-2-oxo-1,3,2-diazaphospholano-3-carbonyl chloride.

Colourless oil; yield: quantitative.

IR V (max., CHCl3) cm-1 1730 and 1 755 (COCI).

c) a-( 1 -Ethyl-2-phenoxy-2-oxo-1 ,3,2-diazaphospholano-3-carbonylamino)benzylpenicillin.

White powder; yield: 71%.

IR v (max., KBr) cm-1: 3350 (NH), 1785 (p-lactam C=O), 1740 (COOH) and 1680 (CONH).

NMR(CDCI3): 1.23 (3H, t, CH2CH3, J = 7.5 Hz), 1.40 and 1.43 (6H, each s, CH3 x 2), 2.75-3.50 (6H, m, CH2 x 3), 4.32 ( 1 H, s, C3H). 5.33-5.80 (3H, m, C5 and CeN and PhCH), 7.25 and 7.30 (1 OH, each s, aromatic protons) and 8.13 (2H, broad s, NH and COOH).

EXAMPLE 15 a) 1 -Ethyl-2-diethylamino-2-oxo-1 ,3,2-diazaphospholane.

Colourless oil; yield: 98%.

NMR (CDCI3) S: 2.00-2.40 (9H, m, CH3 x 3) and 3.80--4.60 (1 OH, m, CH2 x 5).

b) 1 -Ethyl-2-diethylamino-2-oxo-1 ,3 ,2-diazaphospholano-3-carbonyl chloride.

Colourless oil; yield: 87%.

IR V (max., CHCl3) cm-1: 1725 (COCI).

c) a-( 1 -Ethyl-2-diethylamino-2-oxo-1 ,3 ,2-diazaphospholano-3-carbonyla mino)benzylpenicillin.

White powder; yield: 68%.

IR V (max., KBr) cm-1: 1785 (ss-lactam C=O), 1735 (COOH) and 1670 (CONH).

NMR (CDCI3) S: 0.70-1.30 (9H, m, CH3 x 3), 1.48 (6H, s, CH3 x 2), 2.50--3.90 (10H, m, CH2 x 5), 4.30 (1 H, s, C3-H), 5.30-5.85 (3H, m, PhOH, and C5 and C8--H) and 7.30 (5H, s, aromatic protons).

EXAMPLE 16 a) 1-Ethyl-2-methyl-2-oxo-1,3,2-diazaphospholane.

The compound was prepared analogously to the process described in Example 1 Oa and was obtained in a yield of 57%.

NMR (CDC13) #: 1.18 (3H, t, CH3CH2), 1.54 (3H, d, CH3-P, J = 15.8 Hz) 2.80--3.60 (7H, m. NCH2 x 3, NH).

b) a-( 1 -Ethyl-2-methyl-2-oxo- 1 ,3,2-diazaphospholano-3-carbonylamino)benzylpenicillin.

The compound was prepared analogously to the process described in Example 1 Ob and was obtained as a white powder.

IR V (max., KBr) cm-1: 1785 (ss-lactam C=O), 1 730 (COOH) and 1700--1650 (CONH).

NMR (CDCl3) 6: i .00--1.40 (3H, m, CH2CH3), 1.40--1.60 (6H, m, 2C (CH3)2), 1.67 and 1.83 (3H, each d, P-CH3, J = 15.8 Hz) 2.803.50 (6H, m, N-CH2 x 3), 4.35 (1 H, s, C3-H), 5.40--5.80 (3H, m, C5 and QH and PhCH), 6.80-7.10 (1 H, m, NH), 7.39 (5H, s, CeHs) and 8.20-8.50 (1 H, m, NH).

EXAMPLE 17 a) 1 -Ethyl-2-methyl-2,3-dihydro-1 H-1 ,3,2-benzodiazaphosphole 2-oxide.

To a cold solution of 0.98 g (7.2 mmoles of N-ethyl-o-phenylendiamine and 2.91 g (28.8 mmoles) of triethylamine in 20 ml of methylene chloride was added 0.96 g (7.2 mmoles) of methanephosphonic acid dichloride in 20 of methylene chloride dropwise during a period of 30 minutes. The mixture was stirred at room temperature for 1 6 hours, the solvent was evaporated off and thereafter ethyl acetate was added to the residue. The mixture was washed with cold diluted HCI, with an aqueous sodium bicarbonate solution and thereafter with saline, dried over sodium sulfate and thereafter the solvent was evaporated off to give 1 -ethyl-2-methyl-2,3-dihydro-1 H-1 ,3,2-benzodiazaphosphole 2-oxide as a slightly red powder. The product was recrystallized from methylene chloride/diethyl ether.

NMR (CDCl3) S: 1.35 (3H t, CH2CH3), 1.80 (3H, d, POCH2, J = 16.5 Hz), 3.2-3.9 (2H, m, NCH2CH3), 6.4-6.9 (4H, m, aromatic protons) and 7.15(1 H, d, NH, J = 18.5 Hz).

b) 1 -Ethyl-2-methyl-2,3-dihydro-2-oxo- 1 H- 1 ,3,2-benzodiazaphosphole-3-carbonyl chloride.

To a solution of 0.24 g (1.22 mmoles) of 1-ethyl-2-methyl-2,3-dihydro-1H-1,3,2- benzodiazaphosphole 2-oxide and 0.185 g (1.83 mmoles) of triethylamine in 4.5 ml of methylene chloride was added 0.199 g (1.83 mmoles) trimethylchlorosilane and the mixture was cooled in an ice bath and thereafter stirred for 1 hour at the same temperature. The mixture was added dropwise to 2.6 ml of a 10% methylene chloride solution of carbonyl chloride cooled in a dry ice/acetone bath. The mixture was stirred for 1 hour and 30 minutes during which time the temperature was risen gradually to room temperature. Evaporation of the solvent gave 1-ethyl-2-methyl- 2,3-dihydro-2-oxo-1H-1 ,3,2-benzodiazaphosphole-3-carbonyl chloride containing triethylamine hydrochloride.The product was used for the following reaction without isolation.

Rv (max., KBr) cm-1: 1750 (C=O).

c) -(1 -Ethyl-2-methyl-2,3-dihydro-2-oxo- 1 H- 1 ,3,2-benzodiaza phosphole-3 carbonylamino)benzylpenicillin.

To a suspension of 0.426 g (1.22 mmoles) of ampicillin trihydrate and 0.246 g (1.83 mmoles) of triethylamine in 4.5 ml of methylene chloride was added 0.265 g (1.83 mmoles) of trimethylchlorosilane cooled in an ice bath and thereafter the mixture was stirred for 1 hour. To the mixture was added 1 -ethyl-2-methyl-2,3-di hydro-2-oxo-1 H-1 ,3,2-benzodiazaphosphole-3-carbonyl chloride in 5 ml of methylene chloride dropwise cooled in an ice bath and thereafter the mixture was stirred for 4 hours. To 10 ml of water was added the above methylene chloride solution dropwise, keeping the pH value between 7.0 and 7.5 by addition of sodium bicarbonate. The water layer was separated and washed with ethyl acetate and thereafter 5 ml of ethyl acetate was added and the pH value was adjusted to 2.0 with 2 N HCI.The ethyl acetate layer was separated off and the water layer was extracted with ethyl acetate and the combined ethyl acetate phase was washed with saline and dried over sodium sulfate and the solvent was evaporated off to give 0.27 g (yield: 39%, calculated on the basis of 1-ethyl-2-methyl-2,3-dihydro- 1 H-1,3 ,2-benzodiazaphosphole 2-oxide) of a-(1 -ethyl-2- methyl-2,3-dihydro-2-oxo-1 H-1 ,3,2-benzodiazaphosphole-3-ca rbonylamino)benzylpenicillin as a white powder.

IR v (max., KBr) cm~1: 1780 1780(ss-lactam), 1735 (COOH) and 1680 (CONH).

NMR (CDCl3 + 2 drops DMSO-d6) S: 1.10-1.80 (9H, m, C-CH3 x 3), 1.83, 2.09 (3H, d x 2, PCH3, J = 16 Hz), 3.3-4.0 (2H, m, N-CH2-CH3), 5.3-5.8 (3H, m, C5-H, Q-H, PhCH), 6.5-7.1 5 (4H, m, aromatic), 7.1 5-7.70 (5H, m, aromatic protons) and 7.70-8.20 (2H, m, NH x 2).

EXAMPLE 18 The following compounds of the general formula XX can be prepared analogously to the process described in Example 6.

a) &alpha;-(1 Ethyl-2-isopropoxy-2-oxo-1 ,3,2-diazaphospholano-3-carbonylamino)benzylpenicillin.

b) &alpha;-(1-Ethyl-2-cyanomethoxy-2-oxo-1 ,3,2-diazaphospholano-3-ca rbonylamino)benzylpenicillin.

c) a( 1 -Ethyl-2-trifluoromethylthiomethoxy-2-oxo-1,3,2-diazaphospholano-3- carbonylamino)benzylpenicillin.

d) a-( 1 -Ethyl-2-(2-dimethylaminoethoxy)-2-oxo- ,3,2-diazaphospholano-3-carbonyla mi no) benzylpenicillin.

EXAMPLE 19 The following compounds can be prepared analogously to the process described in Example 6, with the proviso that ampicillin trihydrate is substituted by amoxicillin trihydrate: a) a-( 1 -Ethyl-2-methoxy-2-oxo-1 ,3,2-diazaphospholano-3-carbonylamino)-p-hydroxybenzyl- penicillin.

b) a-( 1 -Ethyl-2-ethoxy-2-oxo-l ,3,2-diazaphospholano-3-carbonylamino)-p- hydroxybenzylpenicillin.

c) &alpha;( 1 -Ethyl-2-isopropoxy-2-oxo- 1 3,2-diaza phospholano-3-carbonyla mino)-p-hydroxybenzylpenicillin.

d) a-( 1 -Ethyl-2-cyanomethoxy-2-oxo-l ,3,2-diazaphospholano-3-carbonylamino)-p hydroxybenzylpenicillin.

EXAMPLE 20 The following compounds can be:prepared analogously to the process described in Example 6, with the proviso that 2-(ethylamino)-ethylamine is substituted by 2-(methylamino)ethylamine: a) a-( 1 -Methyl-2-methoxy-2-oxo- ,3,2diazaphospholano-3-carbonylamino)benzylpenicillin.

b) &alpha;-(1-M -Methyl-2-ethoxy-2-oxo-1,3,2-diaza phospholano-3-carbonylamino)benzylpenicillin.

c) &alpha;-( 1-M -Methyl-2-benzyloxy-2-oxo-l ,3,2-diazaphospholano-3-carbonylamino)benzylpenicillin.

d) &alpha;-( 1 -Methyl-2-( p-methoxybenzyloxy)-2-oxo- ,3,2-diazaphospholano-3- carbonylamino)benzylpenicillin.

e) a-( 1 -Methyl-2-phenoxy-2-oxo-l ,3,2diazaphospholano-3-carbonylamino)benzylpenicillin.

f) a-( 1 -Methyl-2-(2,4-dimethoxyphenoxy)-2-oxo- 1,3,2-diazaphospholano-3-carbonyla mino)- benzylpenicillin.

EXAMPLE 21 The following compounds can be prepared analogously to the process described in Example 6, with the proviso that 2-(ethylamino)ethylamine is substituted by 2-(methylamino)propylamine: a) &alpha;-(1 -Methyl-2-methoxy-2-oxo- ,3,2-diazaphosphorinano-3-carbonylamino)benzylpenicillin.

b) &alpha;-( 1 -Methyl-2-ethoxy-2-oxo- 1 ,3,2-diazaphosphorinano-3-carbonyla mino)benzylpenicillin.

c) &alpha;-( 1 -Methyl-2-benzyloxy-2-oxo- 1 ,3,2-diazaphosphorinano-3-carbonylamino(benzylpenicillin.

d) &alpha;-( 1 -Methyl-2-phenoxy-2-oxo- 1 ,3,2-diazaphosphorinano-3-carbonylamino)benzylpenicillin.

e) a-( 1 -Methyl-2-cyanomethoxy-2-oxo- ,3,2-diazaphosphorinano-3-carbonylamino)benzyl- penicillin.

EXAMPLE 22 The following compounds can be prepared analogously to the process described in Example 6, with the proviso that 2-(ethylamino)ethylamine is substituted by 3-(methylamino)propylamine and ampicillin is substituted by amoxicillin: a) a-( 1 -Methyl-2-methoxy-2-oxo- 1 ,3,2-diazaphosphorinano-3-ca rbonylamino)-p-hydroxybenzylpenicillin.

b) a-( 1 -Methyl-2-ethoxy-2-oxo- 1 ,3,2-diaza phosphorinano-3-carbonyla mino)-p-hydroxybenzyl- penicillin.

c) &alpha;( 1 -Methyl-2-benzyloxy-2-oxo- ,3,2-diazaphosphorinano-3-carbonylamino)-p- hydroxybenzylpenicillin.

d) a-( 1 -Methyl-2-phenoxy-2-oxo- 1 ,3,2- diazaphosphorinano-3-carbonylamino)-p-hydroxybenzylpenicillin.

e) a-(1-Methyl-2-(2-dimethylaminoethoxy)-2-oXo-1,3,2-diazaphosphorinano-3-carbonylamino)-p- hydroxybenzylpenicillin.

EXAMPLE 23 The following compounds of the general formula

wherein R21 represents lower alkyl, aryl or benzyl, can be prepared analogously to the process described in Example 10: a) an( 1 -Ethyl-2-ethyl-2-oxo- 1 ,3,2-diaza phosphorinano-3-carbonylamino)benzylpenicillin.

b) a-( 1 -Ethyl-2-trichloromethyl-2-oxo-1 ,3,2-diazaphosphorinano-3-carbonylamino)- benzylpenicillin.

c) a-( 1 -Ethyl-2-trifl uoromethylthiomethyl-2-oxo-1 ,3,2-d iazaphosphorinano-3-carbonylamino)- benzylpenicillin.

d) an( 1 -Ethyl-2-ethoxycarbonylmethyl-2-oxo-1 ,3,2-diazaphosphori nano-3-carbonylamino)- benzylpenicillin.

e) &alpha;-(1-Ethyl-2-benzyl-2-oxo-1,3,2-diazaphosphorinano-3-carbonylamino)benzylpenicillin.

f) a-( 1 -Ethyl-2-phtha limidomethyl-2-oxo- 1 ,3 ,2-diazaphosphorinano-3-ca rbonylamino)benzylpenicillin.

PHARMACOLOGICAL DATA: &alpha;( 1 -Ethyl-2-methoxy-2-oxo- ,3,2-diazaphosphorinano-3-carbonylamino)benzylpenicillin (designated ES 626) and carbenicillin was tested for antibiotic activity in vitro by means of an agar serial dilution test. The following results were obtained::

MIC values in yg/ml Strain/compound Carbici II in ES 626 Proteus morganii 69 0,5 8 Proteus vulgaris 5209 16 8 Proteus vulgaris 16868 16 2 Proteus vulgaris 17028 8 2 Pseudomonas aeroginosa 1999 250 125 Pseudomonas aeroginosa 781 31 16 Pseudomonas aeroginosa 2750 125 31 The LDso value of &alpha; ;-( 1 -ethyl-2-ethoxy-2-oxo- 1,3 ,2-diazaphosphorinano-3-carbonylamino)be nzyl- penicillin intra peritonealfor mice is less than 4 g/kg.

Claims (7)

1. A penicillanic acid derivative of the general formula

wherein R' represents hydrogen or hydroxy, R2 represents hydrogen or a saltforming cation or esterforming group, and R3 represents a group of the general formula

wherein R4 represents hydrogen or optionally substituted lower alkyl, Q represents dimethylene, trimethylene, tetramethylene, optionally substituted o-phenylene or a divalent moiety derived from a 5 or 6 membered heterocyclic compound containing one or two nitrogen atom(s) wherein the two free bonds are situated adjacent to each other, and R5 represents hydroxy, halogen, optionally substituted lower alkyl, optionally substituted aryl, optionally substituted benzyl, optionally substituted lower alkoxy, optionally substituted aryloxy, optionally substituted benzyloxy or a group of the general formula NR6R7, wherein Re and R7 are the same or different and each represents hydrogen, optionally substituted lower alkyl, cycloalkyl, aralkyl or optionally substituted aryl, or Rg and R7 together with the adjacent nitrogen atom from a 5 or 6 membered heterocyclic ring which may contain a further hetero atom selected from the group consisting of nitrogen, oxygen and sulphur, and enantiomers and, when more than one asymmetric carbon atom is present, also diastereoisomers and salts and, when R2 represents a saltforming cation, hydrates thereof.

2. A derivative according to Claim 1 , wherein R4, R6 and R7 are unsubstituted.

3. A derivative according to Claim 1 or 2, wherein Q is a diva lent moiety derived from a 6 membered heterocompound.

4. A derivative according to Claim 1, wherein R3 represents a group wherein Q represents dimethylene ortrimethylene, R4 represents ethyl and R5 represents hydroxy, methyl, methoxy, ethoxy, 2,2,2-trichloroethoxy, phenoxy, benzyloxy or diethylamino, or enantiomer(s), diastereoisomer(s), salt(s) or hydrates(s) thereof.

5. A derivative according to Claim 1, wherein R4 represents methyl, ethyl, propyl or isopropyl, R5 represents hydroxy or methoxy, and Q represents trimethylene.

6. A derivative according to Claim 5, wherein R4 represents ethyl, R5 represents hydroxy or methoxy and Q represents trimethylene.

7. A derivative according to any one of the preceding claims, wherein R1 represents hydrogen and R2 represents sodium.

7. A derivative according to Claim 5 or 6, wherein R1 represents hydrogen and R2 represents sodium.

8. an( 1 -Ethyl-2-methoxy-2-oxo- 1,3,2-diazaphosphortnano-3-carbo nyla mino)benzylpenicillin.

9. A pharmaceutical composition which comprises an effective amount of at least one derivative in accordance with any one of the preceding claims, in association with a suitable physiologically acceptable carrier or excipient.

10. The use of a derivative in accordance with any one of Ciaims 1 to 8 as a therapeutic agent against a microorganism.

11. A use according to Claim 10, in which the microorganism is of the Proteus or Pseudomonas species.

12. The use of a derivative in accordance with any one of Claims 1 to 8 as a growth promotor for ruminant animals.

13. A derivative in accordance with Claim 1, as described in any of foregoing Examples 2 to 23.

14. Any novel feature or combination of features described herein.

New claims or amendments to claims filed on 2nd March 1979.

Superseded claims 2, 3, 6, 7.

New or amended claims:

2. A derivative according to Claim 1, wherein R4, Re and r7 are unsubstituted and wherein the divalent moiety derived from a heterocyclic compound stated for the symbol Q is 6 membered.

3. A derivative according to Claim 1 or 2, wherein ail the substituents stated as being optionally substituted are unsubstituted.

6. A derivative according to Claim 5, wherein R4 represents methyl.