NO750668L - - Google Patents

Description

Mono- and diesters of α-carboxy substituted penicillins.

The present invention relates to new penicillins and methods for their production. The invention also relates to the production of pharmaceutical preparations containing these penicillins and methods for their pharmacological use.

More particularly, the invention relates to new mono- and diesters of α-carboxy substituted penicillins of formula

and their pharmaceutical salts, wherein

R denotes phenyl, thienyl or furyl and R<*> denotes hydrogen. • ;:.

in denotes , hydrogen, alkyl with from 1 ./-^' l to 8 C-atoms, a ryi or aralkyl, which alkyl, aryl and aralkyl groups can be unsubstituted or substituted sed en \ ; . or more groups selected from amino, substituted amino such as V^|^^ methylamino, diethylamino eMer acetamldo, halogen, nltro^•UerV.^^^ . ■ alkoxy with' from 1 to"4 C atoms, -'" -^y-^'^ I is 0 or NH, r3 denotes hydrogen, methyl siler ethyl, R^ is selected from saturated or unsaturated alkyl groups with from 1 to ,/ 8 C atoms, cyclo alkyl groups with from 3 to 7 C atoms, aryl, t-^ fft aralkyl, and heterocyclic groups, where alkyl-j cycloalkyl-,••aryl-t| aralkyl and heterocyclic groups may be unsubstituted*aller."^.l^p substituted with one or more groups selected from amino, aab»-^ §:^, * ■-.■'.We''"■>■ substituted amino such as methyl amino, diethylamino or acetamido, / v,. '' halogen, nitro or alkoxy having from 1 to 4 carbon atoms; provided that R^ bete<g>ner ;'

when f<t>is a hydrogen atom or

Illustrative examples of groups covered by the above definitions and by definitions of groups in the description for-.-v'; others are: - ? f; alkyl: methyl, ethyl, jpropyl, isopropyl, butyl, isobutyl, pentyl, hexyl, heptyl, octyl, 2-ethylhexyl cycloalkyl: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl Alkoxy: methoxy, etbxy, propyloxy, isopropyloxy, butyloxy, isobutoxy ,- - 'halogen: F, Cl, Br *:V: • '> ^••'•-<:>^<>>t aryl: f enyl, iiaphthyl, naphthyloethyl, 5-indanyl ■-, 1%^ aralkyl: benzyl, naphthyylmethyl, 5-indanyl •< heterocyclic groups:

The above illustrative examples show. 1 to the extent they are-^is applicable, all the radicals ;l --RT and other groups that • are mentioned*1 the application within the definition of each group and with restrictions with regard to the number of C atoms that may be prescribed for each group.

The invention included si- also as another drew chemical - v; intermediate products that are new and applicable for the manufacture of K7.' £or- r%^ binders of formula I. v ..... - »+ 4Mi

The compounds of the invention have value for the treatment of infectious diseases in humans or animals, caused 1 , by bacteria. They can be isolated and used as such but also, depending on the presence of acidic or basic groups in the molecule, in the form of salts together with pharmaceutical organic or inorganic acids or bases* Examples of suitable acids are hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, tartaric acid, citric acid and fumaric acid. - Examples of suitable bases are sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, ammonium hydroxide, non-toxic amines such as trialkylamines, including triethylamine, procaine, dibenzylamine, N-beneyl-beta-phenethylamine, 1-ephenamine, N,N-dibenzylethylene-. ;V, diamine, dehydroabiethylamine, N,H -bis-dehydroabiethylethylenediamine, N-(lower)-alkyl-piperidine (e.g. N-ethyl-piperidine) and other bases which<*>have been used for the preparation! of salts with penicillins.

The side chain in the penicillin structure of formula I contains: an asymmetric C atom in the α-position. Depending on the configuration r around this C atom, the compounds will exist in two different :A' diastereoisomeric forms, both of which are biologically active. Likewise, the ester groups can contain asymmetric C atoms, e.g. when RJ ■» CH^or CgH^, which gives rise to different diastereo-isomeric forms which are also all biologically active. It should be understood that the invention includes the pure diastereomers and mixtures thereof.v

It is known that substitution of benzylpenicillin and analogues

compounds 1 a-position on the slde chain with a carboxy group or ' . certain esterified carboxy groups give compounds with general^ structure: .. -.■■,.-•>■ ^

where R has the same meaning as above and R-<*> denotes hydrogen, alkyl^ or aralkyl groups, and these show good antibacterial action, against:J| Gram-positive and Gram-negative bacteria, including Psedomonas aeruginosa (Dutch pstent 6 4^4 3&!»South African patent 67/2804»South African patent 67/6472, Bollard patent 6 805 524, U.S. patent 3 142 673, Dutch patent 6 913 416).

However, these compounds are poorly or only moderately absorbed when given orally, and the iarboxy compounds (II, R** ™ H) are now given by injection. It is an object of the invention to provide esters of these substances which are well absorbed orally and then hydrolyzed in the body to blood and organ levels of the compounds of general formula II which are sufficient for the treatment of the infectious diseases caused by bacteria which are sensitive to penicillins of general;;^ formula II. - : The carboxy group in o-carboxypenicillins (II, R^ » H) is rather unstable and is partially cleaved off during the preparation of the compounds and during storage, forming corresponding non-carboxy-yl penicillins which are less active against gram-negative bacteria , and especially against Ps. aeruginosa. By converting the carboxy group into an ester group, this decomposition is avoided and you get substances that can be easily produced and stored. For i achieve the full antibacterial

effect of α-carboxypenicillins, it is emphasized, however, that one must choose such ester groups which are hydrolysed quickly in vivo during the release of carboxypenicillin. It is an important feature of the invention to provide esters which are stable during production and storage but which, after absorption in the organism, are quickly hydrolysed to high blood and organ levels of ca:eboxypeniciHin. yrt& fcgp

Said compounds of formula I are well tolerated by the organism, produce little side effects and can easily be used in pharmaceutical preparations *' either as such or in the form of their salts, and can be mixed with solid carriers or drench agents and the like. In such preparations, the ratio between the therapeutically active compound and bsere-substances or droying agents is between 1 and 95#» The preparations can either be processed to e.g. pills; pills or dragees or can be given in the form of capsules or filled in bottles. Pharmaceutical, or/?3ni§k$ or inorganic, solid or liquid carriers may be used, of the type suitable for oral el! .is enteral use or for topical use, for the preparation of the preparations, Gelatin, lactose, starch, magnesium stearate, talc, vegetable and animal fat and oil, vegetable gum and polyalkylene glycol and internally known carrier substances for ^ tika can all be used for the production of the preparations in question. " The|% preferred salts of the compounds of the invention are hydrochlorides, r"'-<?>? but salts with other inorganic or organic acids, also antibiotically active acids, can be used, e.g. phosphates, acetates or salts with phenoxymethylpenicillin. Furthermore, the preparation may contain other pharmaceutically active substances which can advantageously be given together with the compound according to the invention for the treatment of infectious diseases. For example, this may apply to certain antibiotics, such as gentamycin.ogyf^i; polymyxin.

When treating bacterial infections in humans, the compounds of the invention can e.g. given in amounts corresponding to 5 to 200 mg/kg/day, preferably 10 to 100 mg/kg/day, in separate doses, HK e.g. two, three or four times daily. D§ is given in dosage units containing e.g. 175, 350» 500 or 1000 mg of active substance.' ^'H^

Examples of preferred compounds according to the invention are: I

Preferred substance classes according to the invention are compounds of formula I where R ex is selected from phenyl, 2- or 3-thienyl or 2- or 3 fu ryi, * /'^i^

is selected from hydrogen, lower-alkyl, benzyl, phenyl, $h indanyl, lower-alkoxycarbonyloxymethyl, l^-lower-alkoxycarbonyloxyethyl, l^-lower-alkoxycarbonyloxy-propyl, lower-alkoxycarbonylaminomethyl, 1'-lower-alkoxycarbonylamino-ethyl, 1 *- lower-alkoxy<y>carbonylamiho-<p>ro<p>yl, , 1<*>-lower-acyloxy-ethyl, 11-lower-acyloxy-propyl, phenokaycarbonyloxymethyl, ,/ 5-indanyloxycarbonyloxyæethyl, l*-phenoxycarbonyloxy-ethylf (5-|S^f^ indanyloxy)carbonyloxy-ethyl, l<*>-b©nzoyloxyethyl, lV<b>^Mo^lox<y>r^^f'propyl, and .--•<-,■ -r-^.- r

R is selected from lower-alkoxycarbonyloxymethyl, l^lower-alkoxycarbonyloxy-ethyl, 1*-lower-alk sycarbonyloxy-propyl, lower- .*•■'' carbonylaminomethyl, l •-lower-ilkoxycarbonylamino-ethyl, l<* >-lower-alkoxycarbonylamino-<p>rop<y>l, f <moxycarbonyloxy-methyl, 5-indanyloxy-carbonyloxymethyl, 1 *-phenoxyki trcarbonyloxy-ethyl, 1' - (5-lndanyloxy) carbonyloxy-ethyl..■ >:■■ : Other preferred classes of substances according to the invention are prepared^ when R<*>" denotes hydrogen and R^ is selected from lower alkoxycarbonyloxymethyl, 1'-lower-alkoxycarbonyloxy-ethyl, 1^-lower-alkoxycarbonyloxy-propyl, la ver e-Alkoxykart onylaainomethyl, 1 * -lower-Alkoxycarbonyl-; "f:araino-ethyl, l^lower-Alkoxycarbcnylamino-propyl, phenoxycarbonyloxy- .: methyl, 5-indanyloxycarbonyloxy-methyl, 1 ?-f enoxycarbonyloxy-ethyl, ^ ^

1r-(5-indanyloxy)carbonyloxyethyl.

Even more preferred classes of substances are obtained when the acyloxy groups in the alkyloxycarbonyloxy, acyloxycarbonyloxy, alkyloxycarbonylamino or acyloxycarbonylamino radicals in R 1 and R 2 have substituents which are amino-met jrlainino or di(lower)alkylamino groups .

The compounds in henholi for the invention can be produced in ( 3 different ways.

' Forward frillinjg; pencl. lln esters

According to this procedure, an activated malonic acid ester derivative III is reacted with an ester of 6-aminopenicillanic acid (6-APA) 17 to form sn penicillin ester V.

When R<2>'R<2>and r<7>' *• fJ constitutes product V a compound•<~^> according to the invention. When R2* or fJ* contain groups that are protected, the jgroups are deprotected, as is known per se, during at least one additional step and give compounds of general formula IA.

In the equations above, the different groups have the following definitions: -f ^

R has the meaning as previously indicated,

R<2>' denotes R<2> as defined previously, or when R2 denotes^ a hydrogen atom or when R contains amino or substituted amino, R denotes this derivative of R 2

-CO-Z denotes a reactive group which can react with an amino group to form an amide residue, e.g. an acid chloride or the latter's functional equivalent. R^* »R?teller denotes, when R^ contains amino or substituted amino, a protected derivative air R^, 7 R' denotes R<2>' is equal

or

presumed that when R? equals

■ • •■ ,->''

E<J>tR<**>" and X have the previously stated meanings. As protecting groups for the carboxyl group, such groups can be used as have previously been used as carboxyl-protecting groups in penicillin synthesis. In particular, the protecting groups can be benzyl . can be removed by treatment with zinc in acetic acid, or via a β-iodethyl, α-p-tolylsulfonylethyl or mono- or dihalobenzyl group which can be removed by treatment with a basic agent such as sodium thiophenolate.

As protecting groups for amino and substituted amino groups, protective groups can be used which can be removed without breaking down the penicillin ring system. Such protecting groups: which are--jffp known in the field are e.g. bejizyloxycarbonyl, o-nitrophenylsulfenyl," • 2-p-tolylsulfonyl-ethoxy-carbonyl, p-trichloroethoxy-carbonyl and 1-methoxycarbonylpropen-2-yl.

The reaction consists in on acylation of an ester of 6-aminopenicillanic acid and can be carried out in the manner described for ■ acylation of other esters of b-aminopenicillanic acid (e.g. as described in French patent 1 57 " 027). The acylating group -CO-Z in -3Tx' compound III can be a jjyre chloride group or a group that functions in the same way, flex, an acid bromide, acid azide, acid anhydride, a mixed anhydridej formed with an inorganic or organic • ■ acid such as an alkylcarboxylic acid, for example isobutyl-carboxylic acid, a carbonic acid, a sulphonic acid and especially an alkoxyformic acid, or can be A a radical prepared from o1 ms! one α-substituted phenylacetic acid and " "• : a carbodiimide or an N,N -carbonyldiimidazole or another compound that reacts in a similar manner.

The reaction can take place in organic solvents such as diethyl ether, tetrahydrofuran, acetone, ethyl acetate, chloroform, methylene chloride, dimethyl formamic, dimethyl sulfoxide or hexamethyl phosphoramide, in water or in strong organic solvents in the presence of organic or inorganic bases such as triethylamine, quinoline, pyridine, N-methylmorpholine, sodium hydroxide, sodium bicarbonate or potassium carbonate. !

The compound of general formula V can be isolated by extraction from the reaction mixture, if necessary after dilution with % water and neutralization. Compounds of general formula V (R<2*>R<2>; R<?>' « R?) to- ; belong to the invention's compounds of general formula I.

Esters of 6-aitiinopenicillanic acid of general formula IV can be prepared by treating 6-APA with compounds R^ - T, where R?<*>;

has the meaning as previously indicated and T denotes halogen or a

functional equivalent of halogen such as an organic sulfonic acid residue. The reaction is preferably carried out in organic solvents. agents such as dimethylformamide or clay dimethylsulphoxide.

Alternatively, 6-acylaminopenicillanic acids containing acyl groups which can be removed without degradation of the perdicillane ring system •'• can be treated with R<?>' - T to form esters of 6-acylaminopenicillanic< acids from which the acyl groups are removed to produce esters of aminopenicillanic acids, i.e. for snow IV. A preferred method consists in reacting a salt such as the sodium, potassium or tetra-alkylammonium salt of benzylpenicillin with R' - T in an organic solvent such as dimethylformamide, dimethylsulfoxide, acetone, chloroform, methylene chloride or hexamethylphosphoraide, or in a mixture of an organic solvent and water, e.g. aqueous acetone or dioxane, for the preparation of the corresponding ester of benzyl--4?penicillin. The phenylacetyl side chain is then removed as described in Dutch patent 6 401 421 or South African patent 67/2927 by treatment with phosphorus pentachloride in the presence of a tertiary organic base !: which gives an imino chloride which oa is added with one alcohol such as propanol, for the production of the corresponding iminoethers which are hydrolysed by the addition of water or alcoholized by the addition of alcohol and form the ester IV. Optionally, the jiide chain can be removed by enzymatic hydrolysis with E. coil acylase as described in French patent 1 576 C27» ;;iff%^

According to yet another process, N-protected 6-aminopenicillanic acid is reacted with R7' 1 -T to produce the corresponding ester, from which the protective groups are removed and give compounds of general formula IV. Examples of protective groups that can be used are the benzyl-oxycarbonyl group, which is removed by catalytic hydrogenation, o-nitro-phenylsulfenyl groups that can be removed by treatment with nucleophilic reagents under acidic pH (Japanese patent 5°5 17&) and trityl-: the group that can be removed by Bild acid hydrolysis.

B. A natural or biosynthetic penicillin with formula

where R°C0 denotes an acyl group on the side chain of a natural or biosynthetic penicillin and M denotes hydrogen or an alkali metal-V

atom such as sodium, potassium, calcium, is etherated by reaction with a compound of formula I

7" I

where R' and I have meanings as previously indicated, after which the eater of formula

which is prepared as follows is reacted with a phosphorus halide in an inert solvent, preferably in the presence of a tertiary amine, to form an iminohalide compound which is reacted with a lower alcohol to give an iminoether derivative which is then reacted with a compound of formula '%v. where R, R<2>' and Z have meanings as indicated above, and the reaction product is then treated with water or an alcohol and forms a for-,, p% bond of formula - V^O

which compound is then converted to a compound of formula I

as described under A above. In this method, the intermediate product in the form of an iminoether compound is directly acylated without isolation of intermediate products. The group R°CO- in the compound of formula VI is an organic

acyl group found in known natural or biosynthetic penicillins. Thus, the group R* can be alkyl, aralkyl or methyl :' :>-" < substituted with a heterocyclic group, or its derivatives. Examples of suitable R° groups are heptyl, phenoxymethyl, 2-thienylmethyl, 2-furylmethyl, and benayl Examples of suitable phosphorus halides are phosphorus pentachloride, phosphorus pentabromide, phosphorus oxychloride, phosphorus

trichloride, etc. Phosphorus pentachloride is preferred. Examples of suitable . alcohols with which the imino halide can be treated are lower alkyl-

alcohols such as methanol, ethanol log n-propanol.

The compounds of formula IX can be converted to any compound of formula V by reaction with compounds of formula j R? 1 where R? has meaning sont earlier and Y denotes halogen or 6 a functionally equivalent group, e.g. a sulfonyl acid residue. When■> Y is halogen, preferably chlorine, bromine or iodine, or when it is enj ;; sulfonic acid residue, such as e.g. a p-tolyl-sulfonyloxy group, the reaction is preferably carried out with a salt such as the sodium, potassium, trialkyl-.. ammonium or tetraalkylammonium salt of compound IX, in an organic solvent such as dimethylformamide, dimethylsulfoxide, acetone, chloroform or methylene chloride, or in a mixture of water

and an organic solvent, such as aqueous dioxane or acetone.

When R2'R<2>andR?' m R^ the product V is a compound according to the invention. Hair R or R' contains groups that are protected, the protective groups are removed as otherwise known, at least; an additional step, during the formation of compounds with a general formula.

IA. ' ■•.:J;:-V'

Penicillins of formula 12, where R 2 * has the meaning previously indicated, are prepared by acylation of 6-amlnopenicillanic acid according to known methods.

The compound of formula X can be reacted with a compound of formula R Jr to form* a compound of formula XI. When R^?-R^ and R<S*>R<8>, the product XI is a compound according to v , the invention. When R^* or f .^'contain groups that are protected,* the protecting groups are removed as otherwise known, in at least one additional step and give compounds of 'general formula I B.

In the above equations, the various radicals have the following definitions: j

R has the meaning as previously indicated,

R is equal to R^ or, when R^ contains amino groups or substituted amino groups, the former denotes a protected derivative

of R<6>.

rR denotes

alkyl with from 1 to 8 C atoms, aryl or aralkyl, which alkyl-, aryl- and aralkyl-.4,/; groups can be unsubstituted or substituted with one or more groups selected from amino» substituted amino, such as methylamino, diethylamino or acetamido, halogen, nitro and alkoxy with from 1 to 4': C atoms,

R is equal to R , or r, when R contains amino groups or substituted amino groups, the former denotes a protected derivative of R 6 , . ■ V.,*^ 8

R denotes

or assuming that R is equal when R8 is equal

-..•'■■ ■.■-■,,. -<&rj-}'-

r<3>, R^" and X have meaning as previously indicated.

Groups mentioned below can be used as protection groups

A.

The reaction conditions described under point C can also be used for the above method. ■.■^■} ;M3.

in

Instead of the activated carboxylic acid derivative of formula III that was used for method A, a ketene derivative of formula XII can be used for the acylation reaction, as described in Belgian patent 726 421.

A ketenoic acid chloride may in formula XIII is reacted with the 6-APA ester of formula XIV after which the compound produced is hydAr* o-ylyzed to form a compound of formula XV. When SC<*>«■ RT. the product XV is a compound according to the invention. ■"Mår%*3-t R^ contains groups that are protected, the protecting groups are removed;^ as otherwise known, in at least one additional step and gives compounds down^# general formula I C.

In the form or the above, the groups R, R^, R^ and X have meanings as previously indicated and R^<*>denotes R^ or, when R^ contains amino groups or protected amino groups, the former forms a • protected derivative of R^.

As protection group sr, the same groups7 • as mentioned under A can be used.

Production of penicillins

in

According to this method, an activated malonester derivative XVI is reacted with a 6-aminopenicillaic acid (6-APA) XVII, to produce ling of a penicillin with fojrmel XVIII, in which formulas R, -CO-Z, J: ^ R^, R^ t R^ and X have meanings as previously indicated.

The reaction conditions are the same as those that can be used for the production of penicillins or acylation of 6-APA. The conditions for the production of the corresponding esters (method A) can partly also be used in connection with the present method.

According to this method, a compound of formula XIX is reacted. where R has the meaning as previously indicated and A represents a protecting group, with a compound of formula

where Y, R-', R\ A<A>' and X have bej't<y>dnin<g>so» previously indicated, forming a compound of formula XI, which is then converted to a compound of formula I D by So replacing the group A with a hydrogen atom, and by replacing the protecting group on R^ as otherwise known.

The group A may be an exi group that has previously been used for carboxyl protection in penicillin synthesis. Especially cat?Al be ;^ benzyl, p-nitro-benzyl or diphenylmethyl, which groups can be removed by catalytic hydrogenation, or A can be an alkyl- or aryl- j"|> group which can be removed by mild alkaline hydrolysis or be a ^ ^i^i p-trichloroethyl group which can be removed by treatment with zlnk I acetic- >?S acid or be an ø-Iodethyl-, 2-p-tolylsulfonylethyl^ or "mono- -^-"'V? or dihalobenzyl group which can be removed by treatment with

basic reagents, such as sodium thiophenolate'

I. Natural or biosynthetic penicillins with formula

where R°GO denotes an acyl group on the side chain of the natural or

biosynthetic penicillin and A has meaning as previously indicated, is reacted with a phosphorus halide in an inert solvent and :' :f-'' preferably in the narvar of a tertiary amine with the formation of an irainp-p^. halide compound which is reacted with a lower alcohol to an iminoether derivative which is then treated with a compound of formula

in

where R, R^, r4, , X and Z have meanings as previously indicated, and the reaction product is treated with water or alcohol to form a compound of formula

which is then converted to a compound of formula I B as described under H above. In this method, the iminoether compound that appears as an intermediate product is directly acylated without isolation of these intermediate products.

The group R°CO- in the compound of formula XXI is an organic acyl group found in known natural or biosynthetic penicillins. The group R° can thus be alkyl, aralkyl or methyl, substituted with a heterocyclic group, or derivatives of the first-mentioned. Examples of suitable groups R are heptyl, phenoxymethyl, 2-thienylmethyl, 2-furylmethyl and beaayl. Examples of suitable phosphorus halides are phosphorus pentachloride^phosphorus pentabromide, phosphorus oxychloride, phosphorus trichlorld etc. Phosphorus pentachloride is preferred. Examples of suitable alcohols as the imino halide can be treated with .'; are lower alkyl alcohols such as methanol, ethanol and n-propanol.

Instead of using the activated carboxylic acid derivative Vr: XVI for this method G, a ketene derivative of formula XXII can be used for the acylation reaction as described in Belgian patent 726 421.

Treatment of carboxypeicillin of formula XXIV, where R has the meaning previously indicated. with a compound Y-CH-X-C-O-R^"

RJ 0

where R-^i R^', X and Y have the previously stated meanings, gives a mixture of two monoesters XVIII and XXV and the diester XXVI. Hair R^'* r4 are all these ester compounds - i. according to the invention with general formula I, and they can be used in the form of the formed mixture. X ^1 When R^"' contains groups that are protected, the protecting groups are removed as otherwise known in at least one additional step.

If one wants the rob connections XVIII, XXV and XXVI, . ^. however, these can be isolated from the &lånding according to known procedures; ways such as extraction, fractional precipitation or crystallization. The best way to prepare the diester XXVI is to treat the carboxypefrcillin XXIV with at least two equivalents • V;

R<3>0 Y-CH-X-C-0-R<4>. RJ 0

In the reaction between the carboxypenicillin and Y-CH-X-C-O-R<4>, the former is used in the form of a dense salt with an inorganic or tertiary organic base, e.g. sodium, potassium, calcium or triethylamine, the salt, and the reaction takes place in organic solvents such as dimethyl-. f formamide, dimethylsulfoxide, acetone, tetrahydrofuran, hecaajoethylphosphoramide or in mixtures of organic solvents and; water, e.g.** aqueous acetone or dioxane.

As described above, the starting material can be used as a salt, e.g. a sodium, potassium, calcium or trialkylmaronium salt, in some of the methods for preparing the compounds in question. :' :±K

4 •■y-t-i:Furthermore, tetraalkylaammonium salts and other analogues can . f-^ :é salts such as salts where the cation has the formula

A XA 2A 3A%

in which A"*" denotes straight-chain or branched alkyl with from 3 to 6%* ;} C atoms, substituted or unsubstituted aryl or substituted or unsubstituted aralkyl, and where A<2>, A^ and A<4>, which may be the same or •'.^^ different, is selected from straight-chain or branched alkyl containing 1 to 6 C atoms, provided that A<2>, A^ and A<4> denote alkyl of 3-6 C atoms when A is equal to alkyl.

Examples of suitable combinations of a\ A2,A?and in the quaternary ammonlum ion can be found in the table below:

Table I. Examples of suitable combinations of groups

A<1>-A4 in the aVaVn<®->ion

When the groups A<*>-A<4> are all different, ' '-^ M dat formed ion contains an asymmetric center and can exist in two enantio-^ more forms» Bpimeric forms can be found if A*t A<2>, A^ and/or/A^ A ^ contains one or more asymmetric C atoms.

Examples of quaternary ammonium ions containing an asymmetric center can be found in Table II:

The above-mentioned use of quaternary salts as starting materials for the production of compounds according to the present-;"

the tetraalkylammonium ion, especially the tetrabutylananonium ion. The preferred solvents are chloroform, methylene chloride and acetone.

The quaternary ctmmonium salt form of the above

starting materials can be prepared by reacting the starting materials in question with a quaternary amrtonium salt of the formula aWaV b°

where A<*>, A<2>, A^ and A<4> have the previously stated meanings and B is

a suitable anion such as HSO^Q , Cl Q or CH^COO Q, while forming a quaternary salt of starting material et. =• V;.^;!<*>

Salts with the above formula containing the B sou anion can be prepared in a known manner 1 analogy with the description ffø.rekelss. e in DHSbe Oe^lfgøli.gsk endpae teeknt sem7p5l^7er 91s« kaAl niiolnluet strB oere are opi pfen innfoerlestern unkykttet erulti-gere.

Example 1. Preparation of 6-(oc-carboxyphenylacetamido)-; J penicillanic acid-a-(ethoxycarbonyloxyethyl)-#

raanoester-naxrlum salt - fv"' <*

a) By method A 1) Chloroethyl ethyl carbonate was prepared by reacting chloromethyl chloroformate (33.7 g, 030 mol) with ethanol (13.8 g, 0.30, mol) A. r in dry ether (500 ral) in norm of pyridine (23.7 g, 0.30 mol). Man ■■■ < continued stirring at room temperature for 3 hours. After filtration, yH^ l and evaporation, the residue was distilled to give a colorless o*4 liquid (33.0 g, 79^, B.P. 14: 48-50°C). 2) To a stirred and ice-cooled suspension of potassium phenylacetate (43»5g»0.25 noi) in dry dimethyl sulphoxide (80 ral) was added .5 •. dropwise chloromethylethyl carbonate \27.7 g, Q20 mol). One continued to : ; stir at room temperature for 18 hours. The mixture was poured into an ice-cooled 0.5 N sodium bicarbonate solution (500 ml) and after stirring for 20 minutes the mixture was extracted with ethyl acetate (3 x 50 ml). The combined organic phase was washed with cold water, dried or anhydrous magnesium sulfate and evaporated. The crude oil

(44>9 g, 94$) was used for the next step.

3) To an agitated solution of S-isopropylcyclohexylamine

(8.4 E» 60 mmol) in dry tetrahydrofuran (60 ml) was added (Ng atmosphere, atmosphere, -78°C) a 1.5 N solution of n-butyllithium in hexane (40 ml, 60 mmol) . After 15 minutes, a solution of the above-mentioned ethoxycarbonyloxymethyl-phenylacetate (13.0 g, 54.5 mmol) in dry tetrahydrofuran (40 ml) was added dropwise over the course of one hour, and a portion of powdered dry ice and continued stirring for 15 minutes. the solution was added dropwise to an ice-cooled solution of 2H hydrochloric acid (100 ml) and after stirring for 15 minutes this mixture was extracted with chloroform (3^75 ml). The combined organic phase was washed with cold water, water (100 ml) added, and pH adjusted to 7#5-•>ir:.;-

with IN sodium bicarbonate solution. The organic phase was washed with water and after washing with chloroform, the combined aqueous phase was added to diethyl ether and the pH adjusted to 1.0 with 2N hydrochloric acid. The aqueous phase was washed with ether and the combined organic phase was washed with water o/v and dried. Evaporation gave a crystalline residue (9.7 g, 63$) which was identified as phenylmalonic acid ethoxycarbonyloxymethyl monoester.

IR spectrum (KBr-sklve) had absorption maximum (cm"'"1") at: 3700-2150 (carboxyl OH): 1755 (ester and carbonate C»0), 1690 (carboxyl 00)". The HMR spectrum in the aterochlorof ona showed absorptions

(p.p.m. ( ) from tetramethylsila1) at 9.50 (a, COOH), 7.33 (s, CgH^),/| 5.79(s, 0CH2O), 4.70 U. Cg<H>^CO), 4.19 (q, OCHgCI^), 1.23 (t, -"'- cf; 0CH2CH3). 4) Phenylraalonic acid The -ethoxycarbonyloxymethyl monoester (1.13 g, 4.0 mmol) was stirred with thionyl chloride (1.67 S, H mmol) at 65°C for one hour and the reaction mixture was evaporated to dryness with dry »;<;> benzene (25 ml) four times.j /tw.

The crude acid chloride (1.20 g, 4.0 mmol) was dissolved in dry methylene chloride (5 mL) and added dropwise to a stirred and ice-cooled solution of 6-aminopenicillac acid benzhydryl ester p-toluenesulfonate (2.22 g, 4.0 mmol) and dry triethylamine (1.01 g, 10.0 mmol) in dry methylene chloride (35 mL). Stirring was continued for 90 minutes at 0°C, then cold water (40 ml) was added and the pH was adjusted to 2.0 with 2H. ^ hydrochloric acid. The organic phase was separated from and washed successively with saturated sodium bicarbonate solution and sodium chloride solution. After drying and evaporation, the residue (2.4 g) was chromatographed on a silica gel column (40 g) prepared in dry benzene. The residue was loaded onto the column in a minimal amount of beneen and eluted with a gradient technique, isopropyl ether-acctone (8:2) being used as the second solvent. The collected frictions were examined by thin layer- ..'.*'. chromatography (TSK) on silica: plates with the same solvent mixture. In this way a white foam (1.30&»505») was isolated from one of the middle fractions in the eluate» This fraction showed only one spot >>;at» "TSK.

IR(KBr): I78O-I74O (p-iLactara-, ester- and carbonate-C-O), I68O Jjl (amide-OO). HMR(CDC13): 7.38 (», 3<C>gHj), 6.94 (s, 0H(C^)z) t 5.79 v,<#>j<:>(s,0CHao), 5»80-5.40 (m, 5-H and 6-H), 4.66 (d, CgH^CHCO), 4.51 (d,3-H), 4.20 (q, OCHgCH^) , 1.60-1.10 (ml OCHgCH^, and cum. CH^). Analysis: Calculated for C^K^yLs (646.73): C 63.14, H 5.30, O 22.27,„• N 4.33, S 4.96. Found; C 63.28, H 4.32, O 22.17, N 4.18, 3 4.86.A 5) The above 6U(α-carboxy^en7lacetamiclo)-penlcillansyTe-> «-(ethoxycarbonyloxynietylj-^^s^ ^y^ry^)-^68^^(1.15 S» 1.8 mmol) was dissolved in a 1:1 mixture of ethyl acetate and ethanol (10 ml) and added a pre-hydrogenated palladium/charcoal catalyst (1.0 g, Fd content 10%) in a mixture of ethanol (5 ml) and water (5 ml) containing sodium bicarbonate (0.15 g»1.8 mmol ). One continued hydrogens none.:/; at atmospheric pressure and room temperature for 2 hours, filtered off the catalyst, evaporated ethanol and ethyl acetate under reduced pressure and washed the resulting mixture with ethyl acetate. Ethyl acetate (10 ml) was added. |?g set and pH adjusted to 2.0 with 2KI hydrochloric acid. The inorganic phase was dried and a 2H solution of sodium 2-ethyl hexanoate (1.0 mL, 2 mmol)* was added. The solution was evaporated to minimum volume, and the sodium salt precipitated with dry ether. The filtered product (0.50 g, $55) showed only one spot by TSK in butanone-pyridine-water-acetic acid (70:15:15:2) as transport system and was identical to, but purer than, “! "''.. H the compound prepared according to method E. >?. IR(KBr): 1780-1740 (8-lactam, ether and carbonate-C-O), 1675 (andd-/^ 00), 1610 (carboxyl-OO). NHR(]Lo); 7.40 (p, Cgi^), 9.80-5.60

(ra, 5-H) 6-R and OCHgO), 4.30 (d, 3-H), 4.10 (q, OCHgCH^), 1.51 (d,

gem, CIU), 1.12 (t, OCHoCfU). Analysis: Calculated for C^Hg^OoNgSNa

(502.49)<:>U 5.58, S,6.38, fla 4.5(8. Found: H 5.45, 3 6.22, Na 4.68. J

The degree of hydrolysis for all the 6-(α-carboxyphenylacetamido)-penicillanic acid derivatives described in this paper was investigated in Sørensen<*>'s buffer solution (3), in 25* human serum

(H) and in 5$ rat serum (R) in the presence of 10$ dimethyl sulfoxide, where the pH in each mixture was adjusted to 6.8. The mixtures were incubated

at 37°C, ffian took samples at different intervals after three hours (33 and H3) and two hours (R2) respectively, samples of which were dripped onto paper and the components of the reaction mixture were separated chromatographically with butanol-ethanol-water (4:1 :1) as solvent systems. The concentration of liberated free 6-(B-carboxyphenylacetamido)-peniclllanate was measured quantitatively by microbiological measurement (Bacillus Subtilis), against stahdard samples and was examined at the same time in the satmne manner. The degree of hydrolysis for the compound described in the present example was: B3 - 24.2*, H3 - 37.5*. -*2 95*)' 'V-b) Ketode G •■;

Phenylmalonic acid ethoxycarbonyloxymethyl monoester chloride (3.0 g,

10 mmol) in dry ether (5 ml) was added to a well-stirred and ice-cooled; solution of sodium 6-aminopenicillanate, prepared by suspending

6-aminopenicillanic acid (3»24 g»15 mmol) in 50% acetone (50 ml) and adjust the pH to 7.0 with 2H sodium hydroxide. During the addition of the acid chloride, the pH was held constant at 7.0 by the addition of '-alkali. r*^$j Kan continued to stir for one hour at 0°C, the organic solvents were distilled off under reduced pressure and the remaining aqueous phase was washed with ether. The pH was adjusted to 4.5 æd 2EJ hydrochloric acid, the precipitate was filtered off and the filtrate acidified to pH 2.2 in nssrvtEr of ether (50 ml). The organic phase was washed with water and extracted by the addition of water (50 mL) and adjusted to pH 7.0 with 2S sodium hydroxide solution. The ether-free aqueous phase was freeze-dried. ^^; a colorless powder (3.1 g, S2r%) which showed a main spot of TSK v-*fi^ (butanone-pyridine-water-acetic acid4systera) beside a minor amount; 6-(α-carboxyphenylacetamido)-disodium penicillanate. This compound was identical to the compound prepared according to method A according to spectral analysis, analytical and pyrolytic data. Example II. Preparation of 6-(α-carboxy-3-thienylacetamido)»| penicillanic acid-o- (ethoxycarbonyloxymethyl)- '■•>■.;; ft^É monoester sodium salt 1) Chloroethyl ethyl carbonate (6.9 g, 50 mmol) was added dropwise to an ice-cooled suspension of potassium 3-tlenyl acetate (10.8 g, 60 mmol) in dry dimethyl sulfoxide (20 mL) ):i and the reaction mixture stirred at room temperature for 20 hours, after which the mixture was worked up in a similar manner as in Example I a. The remaining oil (11.1 g, 9**) was uniform according to TSK artalysis in the system Isopropyl ether- acetone (8:2). I 2) A solution of the above ethoxycarboxyloxymethyl-(3-thianyl)-acetate (7.1 g, 29 mmol) in dry tetrahydrofuran (20 ml) was added dropwise (-78°C, Ng atmosphere) and over the course of one hour added to a solution of lithium N-isopropylcyclohexylamide prepared as in Example Ia by reacting N-isopropylcyclohexylamine (4.2 g, 30 mmol) with en-butyllithium (30 mmol). Powdered dry ice was added, and after 10 minutes the reaction mixture was worked up as in Example!; ^ Ia and gave a product (6.1 g, 73*) jidentified as 3-thienylmalonic acid ethoxy-carbonyloxymethyl monoester. IR(KBr): 3700-2100 (carbonyl-OH),^ I76O (ester and carbonate-C-O), I69O (carboxyl-C-O). HMR(CDCl^): v---;fj> 9.51 (s, COOH), 7.35-7.05 (m, cXs), 5.80 (s, OCHgO), 4.83 (3, C^SCHCO), 4.19 (q, OpJIgCH^), 1J23 (t, OCHgCH^). 3) 3-thienylmalonic acid ethoxy37carbonyloxyethyl monoester chloride bla prepared as in example li from the above acid (1.73 S» 6.0 mmol) and thionyl chloride (1.67 g, 14 ucnol). After working up in the usual way (azeotrope distillation with dry benzene) the crude oil (1.84 g, 6.0 mmol) was dissolved in ether (5 ml) and added dropwise to an ice-cooled 50% acetone solution of sodium 6-amlnopenicillanate, prepared from 6-araino<pe>niellanic acid (1.956»9 mmol) as described in example 1b. Work-up and freeze-drying which in example 1b gave a powder (1, 8 g, 59%) soa showed a main spot in T3K (butanone- ' ;-'\iV.'* pyridine-water-acetic acid-systeqj next to an «inner amount of 6-(a-<x>^<;> vv: carboxy-3-thienyl-acetamido)-ainatriurea-penicillanate IR(KBr): I78O-I74O (B-lactam, ester- ic carbonate-C«*0), I68O-I67O (amide-C-0) , 1610 (carboxyl-OO). NMRfDgS): 7.35-7.05 (m, C₂S), 5.80-5.60 (α, 5-H, 6-H and OHCH₂O), 4.30 (d, 3-H), 4.11 (q,OCt^CH^, ]£0 (d, gem, CH^), 1.12 (t, OCHgCH^). Analysis: Calculated for C^<Hg> jOgN<gS>g<H>atøOS.Sa): K 5.51, S 12.61, Na 4.52. Fuanet: N 5.38, S 12.52, »a 4#62. , ..aJ '2|C. The degree of hydrolysis for this compound was: B3 11.5*»H3 - 32.835, aa - 104&. W Example III. Preparation of S-(α-carboxyphenylacetamido)- penicii; Lanic acid-α-(phenoxycarbonyloxyethyl)- mbnoestor sodium salt 1) Chloromethyl chloroformate (38.7 g, 0.30 mol) in dry ether - r ■ r"* ATI"**' ' "t (150 mL) was added dropwise to a stirred and ice-cooled solution of phenol (28.2 g, 0.30 mol) and pyridine (23.7 g, 0, 30 mol) in dry ether (400 ml). Stirring was continued for 16 hours, the pyridine hydrochloride was filtered off and the filtrate was evaporated, after which the residue (45.3 g) was distilled to give chloromethylphenyl carbonate as a colorless liquid (40.7 g , 73$). B.p. 0.04: 65-68°C. 2) Phenylacetic acid phenoxycarbonyloxymethyl ester was prepared in a similar manner as in Example Ia from chloromethylphenyl carbonate and potassium phenylacetate, and the crude ester was treated at -78°C under nitrogen with lithium-N-isopropylcyclohexyllamd followed by the addition of dry ice as described in Example Ia. After working up in the usual way, euylmalonic acid-phenoxycarbonyloxymethyl monoester was obtained, which was transferred to the 3-oxychloride by treatment with thionyl chloride, :-'/ ' Hp\ excess of the latter was removed by azeotropic distillation with dry benzene 3) A solution of phenylmalonic acid-phenoc sycarbonyloxymethyl- „, monoester chloride (1.74ff. 5.0 mmol) in dry methylene chloride (5 mL) was added to a stirred and ice-cooled | solution of triethylamine (1.21 g, 12.5 mmol) and 6-aminopenicillane urea benzhydryl ester p-toluenesulfonate (2.77 g, 5.0 mmol) in dry methylene chloride (45 ml). After stirring for 90 minutes at 0°C, the reaction mixture was worked up as in Example Ia and gave a yellowish foam (3.34 g) which was chromatographed on a silica gel column (50 g), prepared in dry ben2en. Elution with .,. % ; the gradient technique with isopropylether-acetone (8:2) as the secondary Al^CA solvent gave a major fraction containing 6-(α-carboxy-phenylacetaoido)-penicillanic acid io-(phenoxycarbonyloxymethyl)-3-benz-C'C; ' The hydryldiester (1.84 g, 53$) was isolated as a white foam, which was found to be pure by TSK analysis. IRfSBrh 1780-1740^-(p-lactam-, ether-, and carbonate-C»O), 1675 (amide-C-O). Nrø(CDCl«): 7.45-7.10 (m,4C6H,), 6.95 (sJ(C6H5)2<C>H), 5.80 (s, 0CH2O), 5.80-5.40 ( m, 5-H and 6-H), 4.67 (d, CgH^CHCO), 4.51 (d, 3-H), 1.40 (s, gem.CH-j). Analysis: Calculated for CogH^OgNgS (694.78): C 65*69, ; 5 H 4.93, O 20.73, N 4.03, S 4.64. Found: C 65.75, H 4.96, O 20.62,C-S:;; H4.00, S 4.53... AclSg 4) The diester (1.60 g, I<»>,30 mmol) was hydrogenated over palladium/charcoal as in Example Ca, and the sodium salt (0.82 g, 65* ) became

precipitated in the usual way with natr:Lum-2-ethyl hexanoate. The compound showed only one spot on TSK. v IR(KBr): I78O-I74O (B-lactam, ester and amide-OO), I68O (amide-OO) 1610 (carboxyl-OO). NMR (DgO 2 : 7.45-7.10 (ra, 2 CgH 2 , 5.80-5.60

(m, 5-H, 6-H and 0CH 2 O), 4.3<0>(i, 3-H), 3£0 (d, gem, CH 2 ).

Analysis: Calculated for CgcH^O^SNa (550.52): N 5.9, S 5.83, Na 4.18. Found: N 4.96, S 5.62, Na 4.4é. Degree of hydrolysis: B3~29.2#, H3 j 42.5%, R2 117*.

Example IV. Preparation of 6-(α-carboxyphenylacetamido)- v penicillanic acid o-{acetoxymethyl)-3-(1,-ethoxy-carbonyloxyethyl)-diester

1) Chloromethyl acetate (2.72 g, 25 mmol) was added with stirring to an ice-cooled suspension of potassium phenylacetate (5.23 g, 3 mmol) in dry dimethyl sulfoxide (10 mL) and the mixture was stirred for 16 h . Workup which in example Ia gave acetoxymethylphenylacetate (4.77 g, cC 92%) which was converted to phenylmalonic acid acetoxymethylanoester •C:;.j-(1.97 g» 34*) with lithium-N-isopropylcyclohexylamide and carbon dioxide : as described in previous examples. Treatment of this acid (1.51 g, 6.0 mmol) with thionyl fluoride (1.67 g, 14 mmol) and work-up in the usual manner gave the acid chloride as a crude oil (1.73 g, 6.0 mmol) % which was used directly for the acylation step.

2) Phosphorus pentachloride (16.9 g, 3>mol) was added during

5 minutes and with vigorous stirring to a solution of 6-phenylacet-*%. amido-penicillanic acid 3-(1,-ethoxycarbonyloxyethyl)-ester (13.5&»30 mmol) and N,N-diraethylaniline 112.3 «1»97 mmol) in dry methylene f chloride (135 ml) which was kept at -25°C under dry conditions. nitrogen*:•:• Btier^f •>

. Dry methanol (120 ml) was added over 1.1/2 hours while the temperature was maintained at -25° 30°C. After a further 2.1/2 hours, a solution of sodium chloride (12.0 g) in water (60 ml) was added at -10°C. •"• -/\P;< The mixture was set aside and reached room temperature after which the VI organic phase was separated and evaporated. After repeated treatment:' of the evaporation residue, an oil, with petroleum ether, was then dissolved in

tetrahydrofuran and p-toluenesulfonic acid monohydrate added (5.76"

30 mmol). On addition of ether, crystals of 6-amino-penicillanic acid (1f-etoxycar bonyloxyetyl)-ester-p-toluenesulfonate were obtained:;" fr f . (9.4 g. 62% ). IR(KBr): I785-I745 (8-lactam-, ester- and carbonate-C»0). MR (in deuterium diethyl sulfoxide): 9.50-8.10 (s, very broad, < NH3<+>), 7.36 (q, CI^C^SO^, ^.83 (q, OCN^CH^O), 5.50 (d, 5-H), 5.09 (d, 6- H), 4.41 (s, 3-K), |4.20 (q, OCH^U, 2.É6 (s, Cj^CgH^U, 1.45-1.15 (m, gem, CH ^, OCHlCHgJO and OCHgCH^).Analysis: Calculated for C2D<B>28°9<N>2S2(5°4»59): N 5.55, S 12.71, :^ Found: N 5.36, S 12.52 3) The above compound (3.P3 g, 6.0 mmol) was acylated with phenylmalonic acid acetoxymethyl eonnoester chloride (1.73 g, 6.0 mmol) in the presence of triethylamine (1.52 g , 15 mmol) as described in example Ia. After chromatography on silica gel (60 g) using the usual technique and <1> solvents, the desired product was isolated as a foam (1.56 g, 46*) from one of the middle fractions of the eluate. IR(CHC1«): 1780-1740 (8-lactam-, ester- c g carbonate-OO), 1580 (amide-OO).NMR(CDC13): 7.38 (s, CÆ), 6.77 (q, QCHfCH^O) 5.76 (s, OCHgO), 5.80-5.40 (m, 5-U and 6TH), 4.§Q (d,CgH^CHCO), 4.43 (d, 3- H), 4.20 (q, OCHgCi^), 2.06 (s, Cl^CO), 1.60-1.110 (m, 0CH2CK3, 0CH(CH3)0 and gem;,- CH3). •-' "<A>'<;>Analysis: Calculated for C25H3o°llH2S (566.59), 0 53.00, H 5.34, 0 31.06, . n4.94. s 5.66. I : ■ • ■ - .. Found: C 53.40, H 5.62, 0 31.00, N 4.73, 3 5.58 Degree of hydrolysis: 83 » 3.2$, H3 11.15*, R2 »43.5*.

Example V. Preparation of 6-(a-carboxyphenylacetamido)-penicillar acid-o-(ethoxycarbonyloxymethyl)-3-(1<*>-acetck3ethyl)-diester ulj-jjb_-6-(phenylacetamido)-penicillaric acid-3- (1*-ac ethoxyetyl)»est is (3.786»9.0 mmol) in methylene-k! .oride solution was treated with phosphorus pentachloride (2.07£»99 mmol) at -25° C. in the presence of N»K-, diethylanilln (3.7 a>1» 29.1 mmol), whereupon adding methanol (36 ml) and water as described in Example IV. The crude oil 6-aminopenicillin-;j acid 3-(1'-acetoxyethyl)-ester was dissolved in methylene chloride (80 ml).;^:l, and acylated with phenylmalonic acid tJtoxycarbonyloxymethyl monoester- • ; ; chloride (2.70 g, 9.0 mmol) in the presence of triethylamine (1.36 g, 13.5 mmol) in the same manner as described in Example Ia, Preparation and

chromatography on silica gel 1100 g) was carried out in the usual way, and the desired substance isolated (1.7 g, 35*) from one of the middle fractions of the eluate. IR(CHCl-j): 178O-174O (8-lactam, ester and carbonate-C-O), 1680 (amide-OO). NMR(CDCl 2 ): 7.39 (s, CgHc), 6.87 '•

(q, 0CH(CH3)0), 5.78 (s, OCHg), 5.80-5.40 (m, 5-H and 6-H), 4.6O

(d, CgH^CO), 4.40 (d, 3-H), 4.20 (q, OCR-Ci^), 2.02 (s, (CIUCO)), 1.60-1.10 ( m, gem, CH^, OCHgCH^ and 0CH(CH3)0).

Analysis: Calculated for ^^( Pu^ 3 (566.59): C 53.00, H 5.34, 0 31.06, H 4.94, p 5.66. ] .. Found: C 53.22 , H 5.46, 0 30.62, N 4.82, S 5.62. W'V' Degree of hydrolysis: B3 « 1.2*, H3 - 8.8$, R2 - 38.2 $. ' ! J;

Example VI. Preparation of 6-(α-carboxyphenylacetamido)-penicillanic acid-3-(1<*>-ethoxycarbonyloxyethyl)-monoester sodium salt

a) Method_AA_indirekta^procedure

1) To a solution of 6-aminopenicllanoic acid 3-(1f-ethoxy-carbonyloxyethyl)-ester p-toluenesulfonate (3.24 g, 8j0 mmol) in dry methylene chloride (70 ml) containing triethylamine (2.01 g , 20.0 mmol) was added dropwise, under ice-cooling and stirring, to phenylmalonic acid monobenzyl ester chloride (2.31 g, 8.0 roraol) in dry methylene chloride (10 ml). Stirring was continued for 90 minutes at 0°C and then the mixture was worked up and chromatographed on silica gel (100 g) as described in Example Ia. The diester (3.03 g, 65*) was isolated as

a foam which was pure according to TSK analysis. IRfCHCl^): 178O-174O (8-lactam, ester and carbonate-OO): 1680 (amide-OO). NMR(CDCl ): 7.28 (d, 2 C6H5), 6.77 (q, q2H(jcH3)0), 5.80-5.40 (m, 5-H and 6-H),

IN

5.16 (s, C6H^CK2O), 4.61 (d, CgH^CHCO), 4.42 (d, 3-H), 4.20 (q, OCHgCH^), 1.60-1.10 (m, gem, CH^, OCHgCH- °S OCH(CH3)0). Analysis: Calculated for C^H^O^NgS (584.65): C 59.58, H 5.52, O 24.63,"$ H 4.79, S 5.49. Found: C 59.66 , H 5.60, 0 24.34, N 4.64, 0 5.32 Degree of hydrolysis: B3 1*, H3 - < 1*, R2«47.5*.

2) The diester prepared above (2.92 g, 5.0 mmol) was dissolved in ethanol (10 ml) and added to a prehydrogenated palladium charcoal catalyst (3.0 g, Pd content 10%) in a mixture of ethanol and sodium bicarbonate (0.42 g, 5.0 mmol). The hydrogenation was continued; under normal pressure and room temperature for 2 hours, from raf Purified the catalyst and Evaporated the ethanol at reduced pressure. Ether was added

(25 ml), adjusted to pH 2.2 with 2N hydrochloric acid and the organic phase

was separated and washed with water. Water (25 ml) was added, the pH was adjusted to 7.0 in 2H sodium hydroxide solution and the ether-free aqueous phase was freeze-dried to a white powder (1.60 g, 62*), which was \ pure according to TSS analysis (butanone-pyridine-water-acetic acid- :.' ^ ii system) ♦ IR(KBr): _780p_760 (6-lactam-, ester- and carbonate-CW»,\'' - : \vjf% 1675 (amide-C₂O), 1615 (carboxyl-O₂O). AMR (D 2 O): 7.35 (s, C₂H₂), ^;

6.71 (q, 0CH(CH3)0), 5.70-5.60 (m, 5-H and 6-H), 4.A2 (s, 3-H), 4.15 : (q, OCHgCH- .), 1.50-1.10 (m, gem, CH^, OCHgCH-. and OCHfCK^O). Analysis: Calculated for GggHg^O^Hj3Na (516.51): N 5.42, S 6.21, Ka 4.45.-Found: N 5.36, S 6.16, Na 4.74- Degree of hydrolysis: B3 - 3.5*, H3 •13.5*, R2 83.595....

Crude 6-aminopenicillanic acid 3-(1,-ethoxycarbonyloxyethyl)-ester,

prepared as in Example IV from 6-phenylacetamidopenicillanic acid 3-(1'-ethoxycarbonyloxy9)-ester (2.70 g, 6.0 mmol) was dissolved in 50% acetone (20 ml) and acylated with phenylmalonic acid monochloride (0, 80 g, 4.0 mmol) under the same reaction conditions and preparation method as in example lb. Freeze drying of the ether-free aqueous phase gave the title compound as a powder (0.74 g»36*). This compound was identical to the compound prepared ad Indirekte vei (Via) in terms of spectral analysis, analytical and hydrolytic data, but had :^ lower degree of purity. %.

c) tj2i£^2-^x«.»fi22»2Z2ii' p2

A solution in dry methylene chloride (5 ml) of phenyl-(chloro-

The carbonyD-ketene (0.45 g, 2.5 ran.) prepared from phenylmalonic acid and phosphorus pentachloride (CA. 73, 25451 t, 1970), was added dropwise to *

a stirred solution of 6-aminopenicillane-3-(1'-ethoxycarbonyloxyethyl)-ester-p-toluenesulfonate (1.01 g, 2.0 mmol) in dry methylene chloride (10 mL) containing triethylamine (0, 22 g, 2.2 mmol) at -20°C. Stirring was continued for 90 minutes at 0°C, water (20 ml) was added and the pH was adjusted to 2.2 with <:N hydrochloric acid. The methylene chloride solution was extracted with water (15 ml), pH adjusted to 7»P.m«d;2H.?%.4';-.^ sodium hydroxide solution. The aqueous phase was freeze-dried and gave the title compound as a powder (0,(>6 g, 64*), which by spectra 1 analysis, -,,v. elemental analysis and hydrolytic data was identical to the compound^ produced ad indirect well (Vie), but with less purity. '-/' V^ j'^-rl

d) Method_C

To a stirred and ice-cooled solution of tetrabutylammonium hydrogen sulfate (17.0 g, 50 mmol) in water (25 ml) was added chloroform

(50 mL) and the pH was adjusted to 7.0 with 2N sodium hydroxide solution. 6-(α-carboxyphenylacetamido)-penicillanic acid-α-ben2ylnonoester-v; Potassium salt (25.3 S» 50 mmol) was added portionwise, the organic phase was separated and, after drying with anhydrous magnesium sulfate'■ <Kf*' f', o-chlorodiethyl carbonate (7*6 g, 50 mmol) was added. The solution was left at 40°C for 16 hours

The reaction mixture was evaporated, water (150 ml) added and the product extracted with ether (3x100 ml). The combined organic phase was washed successively with water, saturated sodium bicarbonate solution and water. Drying and evaporation gave an oily residue

(17.4 g) which was chromatographed on silica gel (250 g) according to the usual method. The diester (9.9 g»30»5? ) was isolated as a foam and was identical in all respects to the diester produced indirectly (Via).

2) Hydrogenation of the compound as described previously

in this example gave the title precursor (5.4 g»69$) which was identical to the compound under Via according to spectral analysis, elemental analysis and hydrolysis data.

Example VII. Preparation of 6^(α-carboxy-3-thienylacetamido)-genicillary acid-3-(1'-ethoxycarbonyloxyethyl-^:.a... monoester sodium salt

3-thienylacetic acid (7.1 g, 50 mmol) was stirred with thionyl-. ^, chloride (12.0 g, 100 mmol) at 40°C for one hour and then distilled with dry benzene (4x50 ml). The crude acid chloride (50 mmol) was added dropwise to an ice-cooled solution of aenzyl alcohol (5.4 g, 5° mmol) in pyridine (30 ml) and the mixture was stirred for 18 hours.

The reaction mixture was poured into 50 -1 ice-water and after

acidification with hydrochloric acid, it was extracted with chloroform. The extract was washed with saturated sodium bicarbonate and water, dried and evaporated. The residue was distilled to give pure benzyl-(3-thienyl)acetate (6.75g.58*. Kp0(1, 143-144°t.. A) This ester (4.65g, 20mmol) was converted to 3-thienylmalonic ■ acid monobenzyl ester (4»20 g, 76*) with lithium N-isopropylcyclohexyl amld and carbon dioxide in the same manner as in Example IA.

The acid chloride of the compound (1.92 g, 6.5 mmol) was used for the -:* A*:' acylation of S-aminopenicillanic acid^-t^ethoxycarbonyloxyethyl)- v::-.^-ester-p-toluene-sulfonate (3.28 g»6.5 mmol) by the same method as in example Via. Chromatography on silica gel (69 g) gave pure 6-(oc-carboxy-3-thienylacetamido)-pericillanic acid e-a-ben»yl-3 - (1 * -ethoxy-carbonyloxyethylD-diester (2.23 g, 58*). IRtCHCl^): I78O-I74O (6-lactam, ester and carbonate-OO), I68O-I67O (amide-OO). IrøRfCDCLj), 7.45-7.05 (m, C6H5 and C4H3S), 6.77 (q. QCH(CH-)O), 5.80-5.40 (m, 5-H and 6-H), 5, 16 (s, c6h5c<h>2o), 4.77 (d,C^SCHCO), 4.42 (d, 3-H), 4 4.20 (q, OCHgCH^), 1.60-1, 10 (m, avg. CG^, 0CH2CH3 and 0CH(CH-,)0). Analysis: Calculated for C^H^O-jN-Sg (590.59): C 54.90, H 5.12,

0 24.38, N 4.74, 3 10.86. Found: C 54.85, H 5.22, O 24.54, N 4.62,

S 10.82. Degree of hydrolysis: B3 - < 1*, H3 1*, R2 - 39.8*.

\4) The diester (2.09 g, 3.5 mmol) was hydrogenated over palladium/charcoal (2.0 g) as described in Example Via. The freeze-dried product (1.17 g, 64*) showed only one fleck on TSK (butanone-pyridine-water-acetic acid system). IR: (KBr) I78O-IJ6O (B-lactam, ester and carbonate-C-0), 1675 (amide-OO), 1610 (carboxyl-OO), NMR (D20): 7.35-7.05 (m, C^S), 6.71 (q, 0CH(CH3)0)( 5.70-5.60 (m, 5-H and 6-H), 4.43

(s, 3-H), 415 (q, 0CH2ChT3), 1.50-1.10 (m, gem, CH3, 0CH2CH3 and 0CH(CH3)O). Analysis: Calculated for C20<H>23<0gN>2<S>2??a (522.53): N 5.36, 3 12.27, Na 4.40. Found: N 5.22, S 12.08, Sa 4.62. Degree of hydrolysis: B3 » 4.?£, H3 - 15.3*, R2 - 92.6*.

Example VIII. Preparation of 6-(pt-carboxyphenylacetamido)-penicillin* m acid-o-(ethoxycarbonyloxymethyl)-3-*

(1'-ethoxycarbonyloxy ethyl)-diester

a) Method_Ax_direct_ac}'leration

Crude 6-aminopenicillanic acid 3-(1T-ethoxycarbonyloxy)-ester

prepared as in Example VIb from 6-phenylacetamidopenicillanic acid 3-(l<»>«.ethoxycarbonyloxyethyl)-ester (4»05 g, 9.0 naol) was acylated in

methylene chloride (70 mL), as described in Example Ia, with phenylmalonic acid ethoxycarbonyloxymethyl methyl ester chloride (2.70 g, 9.0 mmol). ;,, - After workup, the crude oil was chromatographed on silica gel (100 g) to give the tltel compound, isolated as a foam (0.75£• 14*)

from one of the medium fractions of the eluate. IR (CHCl-,): 1780-1740 (B-lactam-, ester- and carbonyl-C»0), I685-1675 (amide-C-0), NMRtCDCl^): 7.40(s, c6h5) , 6.3o (q, ocH(k3)o), 5.81 (s, OCHgO), 5.80-5.40

(m, 5-H and 6-H), 4.61 (d, C 6 hIcHC0), 4.41 (d, 3-H), 4.23 (q, QgHgCH^), ; 1.60-1.10 (m, gem, CH 2 , OCH 2 cf 0 and 0CH(CH 3 ) 0 ). Analysis: Calculated for C26<H>32°12N2S (596.67. C 52.34, H 5.41, O 32.18, B 4.70, S 5.38, Found: C 52.44, H 5 .48, 0 31188, N 4.62, 3 5.44. Degree of hydrolysis: B3 - 3.8*. H3 - 12.7*. R2 - 4^.5*.

b) Me^ode^A^addisJons thesis

Phosphorus pentachloride (2, (7 g, 9.9 mmol)) was in the course of 5 -in- v

under vigorous stirring was added to a solution of 6-phenylacetamido-penicillanire-3-(1,-ethoxycarbonyloxyethyl)-ester (4.05 g, 9.0 mmol) and N,N-dimethylaniline (3.7 ml) in dry methylene chloride (40 mL) at -25°C under dry nitrogen. After 1.1/2 hours, dry methanol (36 mL) was added while maintaining the temperature at -25° - -30°C After further

2.1/2 hours, N,N-dimethylaniline (5.4">!)" was added, followed by dropwise addition, over a period of time, of phenylmalonic acid-ethoxycarbonyloxy-:methyl-anoester chloride (3.25 g 10.8 mmol ) in methylene chloride (10 ml). The mixture was stirred for 16 hours at -20° C. Water (27 ml) was added, the pH was adjusted to 2.0 with 2M hydrochloric acid and the organic phase was separated and washed successively with saturated sodium bicarbonate solution and sodium chloride solution. After drying and evaporation, the residue (8.70 g) was chromatographed on silica gel (100 g). The title compound was isolated as a foam (2.30 g, 43^) ^ra a of the medium fractions of the eluate. The compound showed the same spectral data, analytical values and hydrolysis values as the substance (prepared under a).

Example IX. Preparation of 6-(o-carboxy-e-thienylacetamido)-penicillanic acid-α-(ethoxycarbonyloxymethyl)-3-(1•-ethoxycarbonyloxyethyl)-diester

3-thienylmalonic acid ethoxycarbonyloxymethyl monoester chloride

(0.92 g, 3.0 mmol) was reacted with 6-aminopenicillanic acid-3**(1,-^oxy-carbonyloxyethyl D-ester-p-toluenesulfonate (1.51 g»3.0 mmol) in methylene chloride -solution in the presence of triethylamine (0.76 g, 7.5 mmol)

in the same way as described in example Ia. After processing and

chromatography on ailikageL (40 g) the tital compound was isolated

as a foam (0.99 g, 55$) from the middle fractions of an ar eluate. IRfCHCl^): 178O-174O (B-lactam, ester and carbonate-OO), 1680 (amide-OO). NMR(CDCl 2 ): 7.1*5-7.05 (m, C.H^S), 6.78 (q, OCHjCH^O), 5.81 (s, 0CH 2 O, 5.80-5.40 (i , 5-H and 6-H), 4.78 (c^SCHCO), 4.41

(d, 3-H), 4.23 (q, OCHgCH-j), 1.50-1.10, (m, gem. CRy OCHgj^ andH

OCHfCH^O).

Analysis: Calculated for ^4^o°12N2S2 (602.66). C 47.83, H 5.02, O 31.86, N 4.65, S 10.64. (Found: C 47.62, H 4.88, 0 31.58, H 4.44,fe' S 10, 28. Degree of hydrolysis: 83 -2.8*, H3 » 13.6*. R2 - 51, 5*.

Example X. Preparation of 6-(α-Carboxyphenylacetaoido)-peniciLan8yre-α-phenyl-3-(1'-ethoxycarbonyloxy-eth<y>1)-Ester

6-Aminopenicillanic acid 3-(1'-ethoxycarbonyloxyethyl ester p-toluenesulfonate (1.26 g, 2.5 mmol) was acylated in methylene chloride solution, as previously described with Phenylsalonic acid monophenyl ester chloride (0.69 g, 2.5 mmol). Workup and chromatography on <!'^.' f" py silica gel (30 g) afforded the title compound as a foam {0.87 g, 61 *), from one of the middle fractions of the eluate. IRfCHCl^): 1780-1740 (B-lactam, ester and carbonate-OO), I680 (amide-OO). MR(CDC1~): 7.40-7.10 (ra, V 2 C6H5), 6.78 (q, 0CH(CH~)0), 5.80-5.40 (m, 5-H and b-H), 4.60 (d, . |1 CgH^ CHjCO), 4.42 (d, 3-H), 4.22 (q, OCHgCH^), 1.60-1.10 (m, gem. fey;#' Ctiy 0CH2CH-,, and OCH(CH- ,)0).Analysis: Calculated for C^H-^OgNgS (570.64): C 58.94, H 5.30, O 25.23,

S 4.91, S 5.62. Found: C 58.72, H 5.16, O 24.92, N 4.84, S 5.55» Degree of hydrolysis: B3 - cl*, H3 4.2*, R2 - 58.5*.

Example XI. Preparation of (α-carboxyphenylacetamido)-penic: llanoic acid α-(5'-indanyl)-3-(1'-ethoxy-carboyloxyethyl)-diester

6-Aminopenicillanes]xe-3-(1,-ethoxycarbonyloxyethyl)-ester p-toluenesulfonate (2.02 g, 4.0 mmol) was acylated with phenylmalonic acid-,. 5'7indanyl monoester chloride (1.26 g, 4.0 ramole) following the same procedure as in the previous example. After chromatography on silica gel^ -

(50 g) the title compound was isolated as a precipitate (1.51 g»62*) from vAy one of the middle fractions of the eluate. IR(CHCl 10 ): 178O-174O (B-lactam, ester and carbonate-OO), 16«0 (amide-OO). NMRtCDCl^); 7.40-6.90

(sn, C6H5 and indanyl -H), 6.78 (q, OCH(CHo)O), 5.80-5.40 (5-H and 6-H), 4.61 (d, CgH^CHCO), 4.41 (d, 3-H), 4.20 fq, OCHgCH^, 2.86 (t, indanyl-H), 2.30-1.90 (m, indanyl-H), 1.50-1 ,10 (m, average CH^ and

OCH(CH3)0).

Analysis: Calculated for C^H^C^S (610.70): G 60.97, H 5.6l, O 23.58, N 4.59, S 5.25. Found: C b©.52, H 5.38, 0 23.72, N 4.62, 3 5»l8. Degree of hydrolysis: B3 - 1*, H3 12.3*, R2 - 65.8*.

Example XII. PREPARATION OF 6-(α-carboxyphenylacetamido)-penicillanic acid-α,3-(ethoxycarbonyloxymethyl)-diester To a stirred and ice-cooled suspension of 6-(α-carboxyphenylαacetamido)-penicillanic acid-α-(ethoxycarbonyloxymethyl)- monoester sodium salt (1.51 g, 3.0 mmol) in dry dimethylformamide (6.0 ml) containing potassium iodide (0.015) was added dropwise chloromethylethyl carbonate (0.49 E» 3.5 mmol ).

After stirring for 16 minutes at room temperature, the reaction mixture was poured into saturated sodium bicarbonate solution (10 ml) and after stirring for 10 minutes, the mixture was extracted with ether (3*10 ml). The combined organic phase was washed with water, dried and evaporated to an oily residue (1.40 g) which was chromatographed on silica gel (40 g) in the usual manner. The Tite1 compound was isolated as a foam

(0.26 give 15*) from one of the middle fractions of the eluate. IRfCHCl^): 1780-1740 (B-lactam, ester and carbonate-OO), 1680 (amide-OO). NMR(CDCLj): 7.39 (s, C6H5), 5.80 (s, och2o), 5.80-5.40 (m, 5-H and 6-H), 4.60 tr (d, C6H5 £HC0), 4.42 (d, 3-E), 4.21 (q, O-CJIgCH^), 1.50-1.10 (ra, gem..CH^, OCHgCH^). Analysis: Calculated for C25H30°12W23 (582.60): C 51.54, H 5.19, O 32.95, N 4.81, S 5.55. Found: C 51.46, H 5.08, O 32.72, T N 4.72, S 5.36. Degree of hydrolysis: B3 - 13.8*, H3 «29.2*, R2 » 88.2*.

Example XIII. Preparation of 6-(a-carboxyphenylacetamido)-penicillanic acid-3-(ethoxycarbonyloxymethyl)- monoester - sodium salt , , m . m ii 1) To a stirred and Ls-cooled suspension of 6-(α-carboxy-phenylacetaaido)-penicillanic acid-α-benzyl monoester potassium salt (20.2 g, 40 mmol) in dry dimethyl sulfoxide (32.5 ^1) was chlorine added drop by drop? methyl ethyl carbonate (5.5 g, 40 mmol).

After stirring for 16 hours at room temperature, the mixture was poured into an ice-cooled saturated solution of sodium bicarbonate 7 (150 ml). After stirring for 10 minutes, the mixture was extracted with ethyl acetate (3*75 mD) and the organic phase was washed with water, dried and evaporated to an oil (18.5 g) which was chromatographed on silica gel (250 g) with usual 3olvent system. In this way, 6-(α-carboxyphenylacetamido)p»snicillanic acid-o-benzyl-3-(ethoxycarbonyl- j

oxyraetyD diester (9.8 g, ^3^) isolated from the main fraction of the eluate.

IR (CHCl^): I78O-I74O (8-lactam, ester and carbonate-OO), I68O (amide-OO). HM R (CDCl^): 7jf.30 (d, 2 CgH^, 5.78 (s, OCHgO), 5.80-fe 5.40 (m, 5-H and 6-H), 5»l6j( s, CgH^CHgO, 4.60 (d, CgH^CHCO), 4.42 (d, 3-H), 4.22 (q, QCHgCHj, 1.60-1.10 (m, gem. CH^ and OCHgCH^).Analysis: Calculated for CggHLo^HgS (570.64: C 58.98, H 5.30, O 25.23,

N 4.91, S 5.62. Found: C 58.76, H 5.14. 0 25.42, H 4.82, 3 5.46. Degree of hydrolysis: 03 • > 1*, Hp -<1*, R2 - 64.5*.

2) Diestene (9.7 g, 17 mmol) was hydrogenated over palladium/charcoal (10.0 g, Fd content 13*) as described in example Via. The freeze-dried product (6.0 g, 70*) was a white powder showing only one spot on TSK (butanone-pyridine-water-acetic acid system). IR (KBr): I78O-I74O (B-lactam, ester and carbonate-OO), I685-I675 (amide-OO), I615-I605 (carboxyl-OO). NKR (DgO): 7.33 (s, CgH^, 5.70-5.60

(m, 0CH2O, 5-H and 6-H), 4.45 (s, 3-H), 4.17 (q, OCHgCH^, 3^0-1.10 fe (m, gem. CH^ and OCHgCH^).Analysis: Calculated for C^E^OgNgSNa (502.48): N 5.58, 3 6.38, Na 4.58.5 Found: N 5.31, S 6.38, nJ 4.08. : B3 « 18.2*, H3 - 37.5*, R2 - 109*.

Example XIV. Preparation of 6-(α-carboxyphenylacetamido)-penicillanic acid-3-(ethoxycarbonylaainomethyl)-mone ester sodium salt 1) According to the description in Example XIII, 6-(α-carboxyphenylacetaraido)-p<nicillanic acid-α-ben2ylmonoester potassium salt ( 1.87 g, 3.7 mmol) treated with M-(chloromethyl)ethylurethane (0.55 S,

4.0 raraol). After work-up, the evaporation residue was chromatographed on silica gel (40 g), the one diester (0.57 g, 27*) was isolated as a foam from one of the middle fractions of the eluate. IR (CHCl^): 1780-1750 (B-lactam-, and ester-OO) 1720-1630 (amide- and urethane-OO).

NMR (CCCl): 7.30 (d, 2 CgH5), 5.80-5.40 (m, 5-H and 6-H), 5.17 (CgH^O), 4.85-4.55 (m , CgH^CHCO and 0CH2SH), 4.41 (d, 3-H), 4.19

(q, OCHoCH^). 1.60-1.10 (m, average CH^ and OCH^CH^). Analysis: Calculated for<c>2<gH>31<o>8<H>3<s>(569.65): 59.04, H 5.49, 0 22.47. R 7.36, p 5.63. Found: C 58.88, H 5.36, O 22.36, H 7.A2, 3 5.58.

2) The diester (0.40 g, 0.70 mmol) was hydrogenated over palia-v r dium charcoal as described in Example Via and gave, after freeze-drying, the title compound (0.19 g > 54#) soæ a white powder. IR (KBr): 176O-175O (B-lactam and ester-OO), 1720-1680 (amide and urethane-O0), 1610 (carboxyl-OO). NMR (DgO): 7.35 (s, CgH^, 5.70-5.60 (m, 5-H and 6-H), 4.35-4.65 (m, OCjHgNH), 4.43 ( 3, 3-H), 4.15 (q. QCJi2CH3), 1.50-1.10 (m, gem. CH^ and OCHjCH^. Analysis:^Calculated for C2lH24°8R3~ SNa (501.50): R 8.38, S 6.39, |Na 4.58. Found: N 8.28, S 6.18, .0\ Na 4.72. Degree of hydrolysis: B3 - 3.6*, H3»9.8 *, R2 - 73.5*.

Example XV. Preparation of 6-(α-Carboxyphenylacetamido)-penicillinic acid-3-(2*-methylaminoethoxycarbonyl-" oxmetvD- ester salt 1) From chloroauric acid benzyl ester (34.12 g, 0.20 mol) and 2-methylaminoethanol (37, 55 6, Q50 mol) in dry ether (300 ml) prepared J man H-ben«yloxycarbonylmethyl-amino-ethanol (40.36»96*) in the usual way.

2) Chloromethyl chlorophoriate (10.2 g, 72 mmol) in dry ether

(50 ml) was added dropwise to a stirred and ice-cooled solution of

N-benzyloxycarbonylmethylamiroethanol (15.0 g, 72 mmol) and dry pyridine (5.7 g»72 mmol) in dry ether (100 ral). Stirring was continued for 1.5 hours, after which the solution was filtered. The filtrate. '^ was washed with 1H hydrochloric acid (!>0 ml), water (50 ml) and 0.5 N mtrlwa--,. H$?. bicarbonate solution (50 ml. The organic phase was dried and evaporated. The residue was a colorless oil (12.8 g, 59*). NMR (CDCl^): 7.35 Cs, c6H5), 5.73 (s, Cl I :h2o), 5.10 (s, och2c6h5), 4.33 (t, och2-( CH2N), 3.53 It, OCHgCHgN), 2.92 (s, HCH^. v ;

3) To a stirred and Ls-cooled suspension of 6-(α-carbonyl-• x f enylacetaraido)-penicillansy^e-α-benzyl-eanoester sodium salt '--v (25.25 g, 50 mmol) in dry dimethylformamide (80 ml) was added dropwise to chloroethyl-2-N-ban37oxycarbamylmethylaminoethyl carbonate (15.1 g, 50 mmol) in dry dim et ylf orma: Rid (20 ml). After stirring for 16 hours, the reaction mixture was worked up in the usual way and the residue (26.8 g) chromatographed on silica gel (400 g) with isopropyl ether-acetone (7"-3) eo® solvent system. Repeated chromatography from 14.36 isolated compound pi similar manner gave the desired compound as a white foam (10.7 5, 27.8*) which was uniform according to TSK IR (KBr): I785-I765 (B-lactam, ester and carbonate-OO), 1680 (amide-OO). NMR (CDCl 3 ): 7.^0 (s, CgHc), 7.25 («, CgH^, 5.78 (s, > 0CH 2 O), 5.65-5.40 (ra , 5-H and 6-H), 5.16-5.10 (2s, COOCHg C^), 4.6ofe (COCHC6H5), 4.40 (d, 3-H), 4 25 (t, OCHgCHgN) , 3.60 (t, OCHgCH^H), %; 2.95 (NCH^), 2.59 (s, gem. CH,;). Analysis: Calculated for ^HoqO^HoS v (733.81): C 60.55, H 5.36, 0<->24.00, N 5.73, S 4.37-Found: C 60.72, H 5.42, 0 2j.82, N 5.52, S 4.25. 4) N-Bentyloxycarbonsrl-u-benz<y>ester-deri wet prepared above (1.5 J5» 2mraol) was hydrogenated in 50% dioxane (150 al) over palladium/charcoal (3.0 g , Pd content 10*) red room temperature and aoraaW^ff:' pressure. E After 3 hours, the catalyst was filtered off and the pH of the filtrate adjusted to 2.8 with 2H hydrochloric acid. ' The solution was evaporated to a smaller volume (about 20 ml), extracted with ether several times,:;pH^^"^ was adjusted to 4.8. After standing in a refrigerator for several days, a yellow-white powder precipitated which was filtered off and washed with a few drops of cold water. IR l KBr): 2750-2530 (ammonium), 178O-174O.?(8-lactam-, ester- and carbo<tu>t-OQ), 1670 (amide- C-0) •; Degree of hydrolysis: f B3 - 23.8*, H3 - 54.6*, R2 - 87.5*.<;:.>^<:.>!^<;Vv,;>;^ Example XVI. Preparation of 6-(α-carboxyphenylacetamido)- Penicillic acid-α-(ethoxycarbonyloxymethyl)-3-(2'-aminoethoxycarbonyloxymethyl)-diester hydrochloride —~ 1) In the same manner as described in Example XIII, 6-(α-carbocaif enylacetamido)-peni;illanic acid-α- (ethoxycarbonyloxymethyl) monoester sodium salt (4.09.5, 9.2 mmol) treated with clonBOtyl-.-Vj'^-(2-azidoethyl)-carbonate (1.80 g, 10.0 mmol). Chromatography on silica gel (100 g) gave the diester in pure form as a foam (1.416»25*). IR (CHClj): 2150 (azido), 1700-1740 (p-lactam-, ester- and carbonate-;fe C-0) 1680 (amide-C-0). NMR: 7.29 (d, 2 CÆ), 5.80 (s, OCH 2 O), fefe-? 5.80-5.40 (5-H and 6-H), 5.17 (s, C6HcCH2N«), 1.42 (s, gem. CH^j/v-fe^ Analysis: Calculated for CggH^O ^S (6ll765j: C 54.99, H 4.78, 0 23.54, ■ K 11.45, S 5.24, Found: C 55.16, H 4.94, 0 23.42, N 11 .36, S 5.17. 2) The azido diester (920 mg, 1.5 mmol) prepared above, ^ v was hydrogenated in ethyl acetate (55 mL) over palladium/charcoal (0.5 g, 'C\. Pd -content 10*) at room temperature and normal pressure. After 7 hours, the catalyst was filtered off, cold water (10 ml) was added to the filtrate and pH Adjusted to 2.7 with 2N hydrochloric acid, and the organic phase was drained off and washed with water (2x5 ml ). The combined aqueous phase was extracted with isopropyl ether and the aqueous phase was freeze-dried. The white microcrystalline residue is uniform 1 according to TSK. IR (KBr):> 3050 (ammonium), I79O-I775 (8-lactam-, ester and carbonate-C-O) , I685 (amide-C-0), 1510 (ammonium). Degree of hydrolysis: B3 - 28.5*, H3 - 62.5*, Vw;. R2-92.5*.'fe Example XVII. Preparation of 6-(α-carboxyphenylacetamido)- p enicillanic acid 3-(1*-cyclop entyloxycarbonyl-oxyethyl D-ester sodium salt 1) A stream of dry chlorine (220 g, 3.14 »ol) was bubbled through ethyl chloroformate (450 g, 417 mol) at 25-35 °C for 30 hours. During the reaction the mixture was irradiated with white light with a 250 W lamp.^ Fractional distillation of the product (the fractions bl? * examined by chromatography) gave a fraction containing pure (> 95#) α-chloroethyl chloroformate (114 g, 25.6* ). 2) The above compound (50.0 g, 0.35 mol) was reacted with cyclopentanol (30.1 g, 0.35 mol) in the presence of pyridine (27.7 S, 0.35 mol) as described in example Ia. After stirring for 19 hours, the mixture was filtered and the filtrate washed with 2W hydrochloric acid, saturated sodium bicarbonate solution and water. After drying and tnndamplng^^vf; the crude oil (60.3 g, $89) was used directly for the next step. 3) α-chloroethyl cyclopentyl carbonate (2.89 g, 15.0 mmol) was added dropwise to a stirred and ice-cooled suspension of 6-(α-carboxy-phenylacetamido)-penicillanic acid α-benzyl monoester potassium salt (5.07 g»10 .0 mmol) in dry dimethyl sulfoxide (10 mL). After stirring for 18 hours^ the mixture was worked up and chromatographed on silica gel (100 g) "T: as in Example XIII. Pure 6-(α-carboxyphenylacetaraldo)-penicillanic acid-α-benzyl-3-(1t-cyclopentyloxycarbonyloxyethyl)- diester (2.06 g, 33$) was isolated from one of the middle fractions of the eluate. IR (CHC1~): I78O-1740 (6-lactara-, ester- and carbonate-OO) 1680 (aaid-O0). NMR ( CDCl^):7.28 (d,2 C6HJ, 6.78 (q, 0CR(CH3)0), 5.80-5.40 (m, 5-H and 6-H), 5.35-5.05 (m, CgH^O and cyclopenyyl), 4.60 (d, CgH^CHCO), 4.41 (d, 3-H)fe.. 2.00-1.70 (m, cyclopentyl), 1.60 -1.10 (rn, gem. CH^ and OCHfCH^JO). x Analysis: Calculated for O^gH^gO^NgS (624.73): C 6l.53, H 5.8l,>f 0 23, 05, N 4.48, S 5.13. Found: C 61.62, H 5.88, O 22.92, N 4.24, S 5.08. 3) The diesterene (1.87 g, 3, 0 mmol) was hydrogenated and worked up in the same way as in example Via. The freeze-dried product (1.07 &» : 64*) was pure according to TSK (butanone-pyridine-water-acetic acid system).

IR (KBr): I780-I740 (B-lactam-L ester- and carbonate-OO), I68O-I67O (amide-OO), I6IO-I6OO (carboxyl-OO). NKR (DgO): 7.35 (s, C^), 6.71 (q, 0CH(CH3)0), 5.70-5.60 (m, 5-H and 6-H), 5, 35-5.05 (m, cyclo-,5,pentyl), 4.43 (s, 3-H), 2.00-1.70 (m, cyclopentyl), 1.50-1.10 (m, chem. CH^ and 0CH(£H3)0). Analysis: Calculated for C^HggO^NgSNa (556.57)C K 5.03, S 5.76, Na 4.13. Found: N 4.92, S 5.58, Na 4.52. Degree of hydrolysis: B3 » 5.8*, H3 - 15.6«S, R2 - 73.5%.

Example XVIII. Preparation of 6-(α-Carboxyphenylacetamido)-penicillanic acid-α-benzyl-3-(enyloxycarbonyl-oxime) ester According to the description and example XVII, 6-(α-carboxy-phenylacetido)-penicillanic acid-0|-benzyl thioene was -potassium salt (5.07 g, 10.0 mmol) treated with chloromethylbenzyl carbonate (2.01 g, 10.0 mmol) which gave, after chromatography on silica gel (100 g), the pure title compound (2.02 g, 32*) as a foam. IR (CDCl^): 178O-174O (8-lactam, ester and carbonate-C-C), 1680 (amide-O0). NMR (CDCl^): 7.28 (d, 2 CgHc), 5.78 (s, OCH^O), 5.80-5.40 (m, 5-H and 6-H), 5.18 .'.„f (d, 2 CgH^O), 4.61 (d, CgH^dHCO), 4.42 (d, 3-H), 1.42 (s,''gem.'"' CEJ. Analysis: Calculated for C^H^OgNgS (632.70 ): C 62.65, H 5.10, O 22.76, N 4.43, S 5.07. Found: C 62.75, H 5.24, O 22.62, N 4.36,

S 5.02. Degree of hydrolysis: B3 - 1*, H3 - ■ 1*, R2 - 36.5*.

Example XIX. Preparation of 6-(α-carboxyphenylacetaraido)-./: •

Phenylcillanic acid-α-benzyl-3-(2-furfuryloxy-.;

carbonyl .oxymet yl) -di est is '': By the same method as :. examples XVII and XVIII, 6-(α-carboxyphenylacetamidol-penicillic acid-α-benzyl monoester potassium salt (5.07 g, 10.0 mmol) was treated with chloromethyl-(2-furfuryl)-carbonate

(1.91 g, 10.0 mmol). Chromate and: *eparation from the pure diester (1.68 g, 27*) as a foam. IR (CHCl^): 178O-174O (Ø-lactam-, ester- and carbo- ■ nat-OO), 1680 (araid-OO). NMt (CDCl^): 7.50-7.30 (m, CgH^ and C^O),/ 6.55-6.35 (m, C4H3O), 5.78 (s, OCHgO), 5.80 -5.40 (m, 5-H and 6-H),

5.18 (s, C^H^CHgO and C^QCHgOi. Analysis: Calculated for C3iH^o°10N2S (622.66): C 59.80, H 4.85, 0 2:>.70, N 4 ,50, S 5.15 Found: C 60.02,

H 4.78, 0 25.54, N 4.32, 3 5.0^. Degree of hydrolysis: B3 - > 1*, H3 1*,

R2 - 41.5*.

Example XX. Preparation of 6-(α-Carboxyphenyl lac ethamido)-penicillanic acid-3-(Cie-2-methyl-β,3-dioxanyl-5-oxycarbonyloxymethyl)-ethionoester sodium 3-alt

Chloromethyl 5-(cis-2-methyl<r>1,3-dioxanyl)carbonate (1.68 g, 8.0 mmol) was treated with 6-[α-carboxyphenylacetamido)-penicillanic acid α-benzyl monoester -potassium salt (4.05 g, 8.0 mmol) as described previously. Chromatography gave the pure diester (1.44 g»28*) as a foam. IR (CHCl^): I78O-I746 (B-lactam, ester and carbonate-OO), I69O-I68O (amide-OO). NMR (CDCLO: 7.29 (d, 2 CgHj), 5.80 (s, OCHgO), 5.80-5.40 (m, 5-H and 6-H), 5.19 (s, CgH^ CHgO), 4.69 (q, OCH (CHgOjg- >CH,CHo), 4.65-4.40 (m, 3-H, OCJiy(CH2O)2>CHCH3 and CgH^CHCO, 4.20-3.90 (ra , XH<(CJi20)2>CHCH3), 1.60-1.10 (m, cum. CH^ and OCH^CHgOj^CHCH^). Itnalysis: Calculated ^or C^h^OjjN^S (642.69 ): C 57.94. H 5.33" 0 27.38fe N 4.36, S 4.99. Found: C 57.82, H 5.24. 0 27.42, N 4.30, S 4.72 .#J

The diester (1.22 g, 1.9 pmol) was hydrogenated on palladium/charcoal in the usual manner to give, after freeze-drying, the title compound (0.72 g, 66*) as a white powder. IR (KBr): I78O-I740 (is-lactam-, Ester- and carbonate-OO) I69O-I680

(amide-OO),1620-1600 (carboxyl-OO).HMR(DgO):7.35 (s, .C^5)fe§S|^ 5.80-5.60 (m, OCH^O, 5-H and 6-H), 4.70 (q, OCH^tCHgOj^CHCH^) ^4.65-feS: 4.35 (m, 3-H, CgH^CO and 0CH.<(]H 2 O) 2>CHCH3), 4.20-3.90 (m, OCUr. (£Egb)2-VCHCJ3). Analysis: Calculated for C 2 H 2 O-^NgSNa (574.54): » 4.88, 3 5.58, Na 4.00. Found: N 4.66, S 5.42, Na 4jl8. Degree of hydrolysis: B3 - 7.6*,

H3«21.5*, R2«88.5*.

Example XXI. Preparation of 6-(α-carboxyphenylacetamido)- r .

penicillin acid-a ,3- (1 * -ethoxycarbonyloxyethyl).- ? diester > ■ A suspension of sodium bicarbonate (15.1 g, 180 mmol) and 6-(α-carboxyphenylacetamido)-penicillanic acid-3-(11-ethoxycarbonyloxy-ethyl)-monoester sodium 3alt (15.5 g* 3° mmol) a-chlorodiethyl carbonate (13.7 g, 90 mmol) was added dropwise to 50* dioxane (30 ml) and with stirring and I-air cooling.

The reaction mixture was stirred for 64 hours, then filtered and chloroform (100 ml) was added to the filtrate. The organic phase was separated and washed with water saturated sodium bicarbonate solution and water, one after the other. After evaporation of the residue, it was kept under high vacuum (0.01 mmfy;) for 12 hours to remove the rest of the dioxane and the α-chlorodiethyl carbonate. Chromatography on silica gel (150 g) with usual. Solvent syst<sm gave the title compound (1.50 g, 3.2*) in pure form, isolated as a foam from one of the two main fractions of the eluate. The second major fraction contained 6-(phenylacetaraido)-penicillanic acid-3-(1,-ethoxycarbonyloxyethyl) ether. IR (CHCl^): I78O-I74O (B-lactam^,-, ester- and carbonate-OO), I69O-IÉ7O (aæid-C«0). NMR (CDCl 3 ): 7.39

(3, C6H5), 6.78 (q, 0CH(CH3)0), 5.80-5.40 (ra, 5-H and 6-H), 4.60 H (d, CgH^CHCO), 4.41 (d, 3-H), 4.20 (q, OCHgCH,^, 1.60-1.10 (m, gem.fe CH^, 0CH2CH3and 0CH(CH3)0). Analysis: Calculated for C27<H>3<4>°12<N>2S

(610.65): C 53.11. H 5.61»0 31.44, N 4.59, S 5.25. Found: C 53.25, H 5.72, O 31.28, N 4.46, S 5.15. Degree of hydrolysis: B3 1*, H3 - 8.5*,; R2 - 32.5*.

Example XXII. Preparation of 6-(α-carboxyphenylacetaroido)-penicijlans<y>re-α-(1'-ethoxycarbonyloxyethyl)-monoester sodium salt "'VC

a) Method^G

1) To a stirred and ice-cooled suspension of phenylmalonic acid bionobenzyl ester (40.5 g, 0.15 noi) and sodium bicarbonate (88.2 g, 1.05 mol) in 50* dioxane (150 ml) was added dropwise a -chlorodethyl carbonate (68.6 g, 0.45 mol).. then stirring was continued at room temperature for 64 hours. ".■' r?:. y

The precipitate was filtered off, and chloroform (500 ml) was added to the filtrate. The organic case was drained off and washed with water, saturated sodium bicarbonate solution and water, successively. After evaporation, the residue (89.5 g) was kept under high vacuum (0.01 mm Hg) for 16 hours to remove remaining α-chlorodiethyl carbonate and dioxane. The evaporation residue (46.3 g) was chromatographed on a silica gel column. .

(300 g)» prepared in carbon tetrachloride. The compound was affixed ■'■^ u:y. ;-without dilution and eluted with a gradient technique with chloroform as second solvent.

As a second fraction, phenylrealonic acid benzyltri<*>-ethoxycarbonyloxyethyl-diester (9.7 g»16.5*) was obtained as a colorless oil., . <

2) The diester (9.5 g»24.6 mmol) was dissolved in ethyl acetate

(100 ml) and hydrogenated at room temperature and atmospheric pressure over palladium charcoal (4.25 g, Pd content 5*) until one equivalent of hydrogen was absorbed. The catalyst was filtered off and the filtrate evaporated to give phenylalonic acid diethoxycarbonyloxyethyl monoester (5.8 g, 80%) as a colorless syrup. in (film): 3500-3200 (hydroxyl), I76O-I74O (ester- and carbonate-OsO), , .. 1600 (carboxyl-C-O). NMR (CVZlj) : 10.20 (s, COOH), 7.32 (s, CgH^, 6.73(q, OCHfCH^O), 4.65 (s, :6H^CHCO), 4.12( q, OCHgCK^), 1.60-1.10 (m, OCHgCH^ and 0CH(CH3)O).

3' Phenylmalonic acid di-ethoxycarbonyloxyethyl monoester chloride (3.15 g, 10 mmol), obtained from the corresponding acid (2.96 g, 10 mmol) as described above, was used for the acylation of sodium -6-aminopenicillanate in the same way m as described in example Ib. The freeze-dried product (3.10 g, l>0#) showed a major spot on TSK (butanone-pyridine-water-acetic acid system), alongside a minor amount of di-sodium 6-(α-carboxyphenylacetÅmido)-penicillanate . IR (KBr): 178O-174O (p-lactam, ester and carbonate-loo), 1680 (amide-OO), 1610 (carboxyl- - C-0), mR(D20: 7.40 (s, C5H5 )j, 6.70 (q, OCHfCH^O), 5.70-5.6"0

(m, 5-H and 6-H),'4.30 (s, 3-H) ,| 4^2 (q, OCHgCH^, 1.60-1.10 (m, OCHgCH^, 0CH(CH3)0 and gem. CH^)L Analysis: Calculated for CggH^OgNgSNa (516.51): N 5, 42, S 6.21, Na 4.45."* Found: N 5.28, S 6.12, Na 4.5&. Degree of hydrolysis: B3 - 4.8*, H3 - 13.8*, R2 -< 8>5.3^. /' m J,

b) Method_H j r=?i-"# \

1) A suspension of 6-(α-carboxyphenylacetamido)-penicillanic acid-3-benzbydryl monoester sodium salt (17.0 g, 30 mmol), (prepared by acylation of 6-aminopenicyllans<y>re-3-benzh<y>dr <y>le3ter-p-toluene-'jfe sulfonate with phenylmalonic acid monochloride) and sodium bicarbonate (15.1 g,^ 180 mmol) in 50$ dioxane (30 ral) was treated with α-chlorodiethyl carbonate (13.7&» 90 mmol) in a similar manner as described in Example XXI. After work-up and chromatography on silica gel, pure (6-(α-carboxyphenylacetamido)-penicillanire-α-(1*-ethoxycarbonyloxy-v ethyl)-3-benzhydryl-diester (j.74 g, 8.8*) was obtained which was isolated from the eluate's second main fraction. CDCLj): jM (d, 3 CgH^), 6.95 (s, (CgH^gCHj, 6.78 (q, 0CH(CH3)0), 5.80-5.40 (m, 5-H and 6-H), 4.66 (d, CgH^CHCO), 4.51 (d, 3.H), 4.20 (q, OCH^H^), 1.60-1.10 (m, OCHgCH ^.OCHfCH^O and gem.CH^).Analysis: Calculated for ^^H^OgNgS (660.76), C 63.62,;.H 5.49, O 21.79, N 4.24, S 4.85. Found: C 63.78, H 5.62, O 21.68, fe N4.16, S 4.32. 2) The diester (1.65 is, 2.5 mmol) was hydrogenated on palladium/ • coal according to the same procedure and work-up method as described in example 1 Ia. The product (0.92 g, 71$) identical to the compound prepared by method E according to spectral analysis, elemental analysis and hydrolytic data, less pure than the latter.

Example XXIII. Preparation of 6-(α-Carboxyphenylacetamido)-penlcllanoic acid-α-(51-Indanyloxycarbonyloxymethyl)~monoester sodium salt Natrlum-6-arainopenic illanate in 50% acetone solution was '• . acylated with phenylmalonic acid-(*'-indahyloxycarbonyloxymethyl) monoester chloride (1.17 g, 3.0 mmol) according to the same method and work-up process as in example Ib. The freeze-dried product (1.05 g, 59%) showed a \'-~'"T main spot on TSK (butanone-pyridine-water-acetic acid system) next to a minor amount of disodium 6-(α-carboxyphenylacetamido) -penicillanate IR(KBr): I78O-I74O (B-lactam-, ester and carbonate-OO), I69O-I68O

(amide-C=0), I610-1600 (carboxyl-C=0). NMR (D 2 O: , 7 , 40-6.90 (m,'

CgH^ and indanyl-H), 5.80-5.50 (m, 0CH2O, 5-H and 6-H), 4.30 (s, 3-H), 2.89 (t, indanyl-H> , 2.30-1.90 (m, indanyl-H), 1.50 (s, gem. CH^). Analysis: Calculated for C28H27<OgN>2SNa (590.59): N 4.74, S 5 ,43, Na 3.89-Found: N 4.66, S 5.32, Na 4.12 Degree of hydrolysis: B3 = 17.2%,

H3=35.4%, R2 = 83.6%.

Example XXIV. Preparation of sodium ct-carboxybenzylpenicillinate a-(isopropoxy-carbonyl lox yrne ty 1) mon oe st er^ .__

Method A

a) Phosphorus pentachloride (2.3 g, 11.0 mmol) was added over 5 min with vigorous stirring to a solution of

6-phenylacetamidopenicillanic acid benzyl ester (4.25 g, 10 mmol) and N,N-dimethylaniline (4.12 mL, 32.5 mmol) in dry methylene chloride

(40 mL) at -25°C under dry nitrogen. After 1.5 h, dry methanol (40 mL, 1.0 mol) was added while maintaining the temperature at -25 to -30°C. After an additional 2.5 h, N,N-dimethylaniline (6.86 mL, 54.1 mmol) was added, followed by the dropwise addition over 30 min of phenylmalonic acid and isopropoxycarbonyloxymethyl monoester chloride (3.8 l g, 11.5 mmol) in methylene chloride. The mixture was then stirred for 16 hours at -20°C. Water (40 ml) was added, the pH was adjusted to 2.0 with 2N hydrochloric acid and the organic phase was separated and washed successively with saturated sodium bicarbonate solution and sodium chloride solution. After drying and evaporation, the residue (7.8 g) was chromatographed on silica gel (160 g). The benzyl ester of the desired compound was isolated as a foam (3.8 g, 65.2%) from one of the middle fractions in the eluate. IR (CHCl 2 ): 1780-1740 (B-lactam, ester and carbonate C=0); 1685 (amide C=0).

NMR (CDCl-j): 7.40 (s, 2 CgH 2 ; 5.80 (s, 0CH 2 O); 5.80-5.50 (m, 5-H

and 6-H.); 5.20 (s, C 6 H 5 -CH 2 ); 5.10-4.60 (m, OCH(CH3)2), 4.65 (d, Cg+^-CHCO), 50 (d-3-H); 1.50 (d, gem CH 3 ); 1.30 (d, CH(CH 3 ) 2 ). Analysis: Calculated for C2gH32<N>20gS (584.67). C 59.57; H 5.52;"

N 4.79;024.63; pp. 5.48.

Found: C 59.26; H 5.74; N 4.65; 0 24.76; S 5.44.

b) The diester obtained above (2.92 g, 5 mmol) was dissolved in ethyl acetate (10 ml) and added to a prehydrogenated

palladium-charcoal catalyst (3.0 g, Pd content 10%). Hydrogenation

was complete after 15 minutes and then the catalyst was filtered off, water was added (10 ml) and the pH adjusted to 6.8 with N sodium bicarbonate solution. The aqueous phase was extracted with isopropyl ether (15 mL) and the isopropyl ether-free aqueous phase was freeze-dried to a white powder (1.76 g, 68.5%) which was pure by TLC analysis (butanone-pyridine-water-acetic acid system).

IR (KBr) 1780-1760 (B-lactam, ester and carbonate C=0); 1675 (amide C=0); 1610 (carboxyl C=0). NMR (D 2 O); 7.40 (s, C₂H₂); 5.80-5.60 (m, 5-H, 6-H and OHCH 2 O)-, 4.40 (s, C<g>H^HCO); 4.30 (d, 3-H); 1.60 (d, gem CH 3 ); 1.30 (d, CH(CH 3 ) 2 ).

Analysis: Calculated for C^H^O^SNa (516.51); C 51.1-6; H 4.88;

N 5.42; S 6.21; Na 4.45-

Found: C 50.02; H 4.72; N 5.26; S 6.10; N 4.58.

Degree of hydrolysis: B3 = 18.2%; H3.= 42.5%; R2 = 97%.

Claims (1)

Procedure for the preparation of a compound of the formula: and therapeutically acceptable salts thereof, wherein R is a phenyl-, thienyl or furyl group; R 1 is a hydrogen atom or a methyl group; R<4> is selected from saturated or unsaturated alkyl groups with 1-8 carbon atoms, cycloalkyl groups with 3-7 carbon atoms, a phenyl group, a benzyl group, an indanyl group, a thienyl group, a furyl group, a pyridyl group and a 2-methyl-1,3 ~ dioxanyl group, where all R 4 groups can be unsubstituted or substituted with one or more groups selected from amino groups, substituted amino groups such as methylamino, diethylamino or acetamido groups. halogen and nitro, characterized in that a natural or biosynthetic penicillin with the formula: where R°-CO- is the acyl group in the side chain of the natural or biosynthetic penicillin, is reacted with phosphorus halide in an inert solvent; the iminohalide compound thus formed is reacted with a lower alcohol and the iminoether thus obtained is reacted with a compound of the formula: "5 .. 4' 4 where R, R^ and X have the meaning given above, R is R as defined above, or when R 4 contains amino or substituted amino groups, a protected derivative of R , A is a protecting group and - CO-Z is a reactive group that can react with an amino group to form an amide residue, after which the protecting groups in the compound of the formula: is cleaved to form a compound of formula ID; after which, if desired, a compound of formula ID thus obtained is converted into a therapeutically acceptable salt thereof.