PH26810A - Penicillanic acid derivatives composition and a process for the preparation thereof - Google Patents

Description

a8"

This invention relates to penicillanic acid derivatives and the preparation thereof. More particularly, the invention relates to penicillanic acid 1,1-dioxide, and esters thereof readily hydrolyzable in vivo, which are useful as antibacterial agente, and also for enhancing the } effectiveness of several p-lactam antibiotics against many p-lactamase producing bacteria; and also to derivatives of penicillanic acid 1,1-dioxide having the carboxy protecting group, which are useful intermediates for the preparation of ) . penicillanic acid 1,1-dioxide. The invention is : also concerned with penicillanic acid 1l-oxides and certain esters thereof which are useful intermediates for the preparation of penicillanic : acid 1,1-dioxide and ite esters.

Well-known and widely used antibacterial agente are the so-called p-lactam antibiotics.

These compounds are characterizad by a nucleus ’ consisting of a 2-azetidinone (P-lactam) ring fused to either a thiazolidine or a dihydro-8,3-thiazine ring. When the nucleus contains a thiazolidine > ring, the compounds are designated penicillins, ] : 25 whereas wher the nucleus contains a dihydrothiazine ring, the compounds are designated as . : cephalosporins. Typical examples of penicilline which are commonly used in clinical practice are . benzylpenicillin (penicillin @), , 5 BAD ORIGINAL 9

. | 087 phenoxymethylpenicillin (penicillin V), ampicillin and carbenlicillin; typilcal examples of common. cephalosporins are cephalothin, cephalexin and cefazolin.

However, despite the wide ure and wide acceptance of Lhe p- lactam antiblotica as valuable ’ chemotherapeutic agents, they puffer from the major drawback that certain members are not active against certain microorganisms. Apparently, this resistance of a particular microorganism to a given p-1lactam antibiotic occurs because the microorganism produces a B- lactamase, which 1s an enzyme which cleaves the p-lactam ring of penicillins and cephalosporins to glve products which are devoid of antibacterial 16 "activity. However, certain substances have the ability to inhibit p-lactamases, and when a P- lactamase inhibitor is used in combination with a ponicillin or cephalosporin it can increase or enhance the antlbacterial effectiveness of the penicillin or cephalosporinagainat certain microorganisms. It is considered that there is an i enhancement. of antibacterial effectiveness when the antibacterial activity of a combination of a B- lactamuse inhibiting substance and a p-lactam 26 antibiotic is significantly greater than the sum of the antibacterial activities of the individual components. 3 leap ORIGINAL 2 :

, AL . The present invention provides certain new penicillin compounds which are useful as : antibacterial asgonts and are also potent inhibitors of microbial b lactamasen. These new penicillin 6 compounds are penicillanic acid 1,1-dioxide, and esters thereof readily hydrolyzable in vivo.

The invention also provides derivatives of penicillanic acid 1,1-dioxide having a carboxy protecting group, which are umeful as intermediates for the preparation of penicillanic acid 1,1- dioxide; and the invention further provides ’ penicillanic acid l-oxides and certain esters thereof, which are alpo useful as intermediates for . penicillanic acid 1,1-dioxide. 16 According to the invention, there is provided a novel penicillanic acid 1,1-dioxide . derivative of the formula: 0 0 wo / oH 3 ALS? > 1 . ‘OH

To . : 6 5 27 3 ’ * 2 r 5 3 . - cn | 7 “ yt 0 - ‘ “coor} : : --- (1) 3 and the pharmaceutically-acceptable salts of those compound wherein rR! ia hydrogen, wherein rR! is : hydrogen, an estear- forming residue readily 4

J i

$10 hydrolyzable in vivo, or a conventional penicillin carboxy-protecting group. The term “ester—forming residues readily hydrolyzable in viva" as used herein refers to non-toxic ester residues which are 8 rapidly cleaved in mammalian blood or tissue to release the corresponding free acid (i.e., the compound of formala 1, wherein rl is hydrogen).

Typical examples of such readily hydrolyzable ester- ’ forming residues represented by rl are 10 alkanoyloxymethyl having from 3 to 8 carbon atoms, 1-(alkanoyloxy)ethyl having from 4 to 9 carbon atoms, l-methyl-1-(alkanoyloxy)ethyl having from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from ro 3 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl : 15 having from 4 to 7 carbon atoms, 1l-methyl-1- © (alkoxycarbonyloxy)ethyl having from 6 to 8 carbon atoms, 3-phthalidyl, 4-crotonolactonyl and y- butyrolacton—-4-yl. ’ The compounds of the formula I, wherein rl 20 ia hydrogen or an ester forming vesidus readily hydrolyzable jin viva, are useful as antibacterial agents and for enhancing the anti-bacterial activity of f-1actam antibioticna. Hereinafter, these antibacterial compounds are designated by the 26 formmla IA.

The compounds of the formula I, wherein rt is a penicillin carhoxy protecting group, are useful as chemical juntermediates to the compound of the i formula IA. Typical carboxy protecting groups are benzyl and substituted benzyl, 8.g., 4-nitrobenzyl.

The present invention also provides 8 process for the preparation of a penicillanic acid 1,1-dioxide derivative of the forsmla: 0 0

BH 7, / Oily . ’ . 7 IN on may

N

Ce “, 0 ! COOR ...IA i and pharmaceutically-acceptable aalts thereof, wherein R is hydrogen or an ester—forming residue readily hydrolyzable in vivo, characterized by reacting a compound of the formulae: 5s on " ’ % o 3 ~ 5 wn CH

X =” INQ

CHy H of M “, N ”, 3 or ; ’ coor? 0 “2 coor! (11) (111)

H }

CH

: % ® RN 3

N

Cc

Hs oo +s “, , 0 coor? (Iv) 8 a

Jf wherein Rr! ia hydrogen, an ester forming residue readily hydrolyzable in vivo or a conventional penicillin carboxy- protecting group, with an oxidizing agent until oxidation to the corresponding 1,1-dioxide of the formula: 0 / ”, “ ow CH,

TP vo NT, 0 nooR? (ID wherein r! in as defined above, is substantially complete, and, if necessary, removing the carboxy protecting group, to form the deaired product of

Co 10 formula IA and, if desired, torming a CL . pharmaceutically acceptable salt by reacting a compound of formula IA, wherein R is hydrogen, with . a bane. to

The intermediates of formulae (11) and (111) above are novel compoundsg and, accordingly, the invention also provides a compound of the } formula: 0

H, : o CHq

Tr | “CH, : / " “ond 0 or LIL)

vt °

H, { 0 . 7, Sc ™ My - : | | | ~ cH,

HN

: 0? “coon ...(111) and the salts thereof, wherein rl is as defined : } above. .

The novel compounda of formulae I, II and

TIT above are roferred to harein apg derivatives of penicillanic acid, which is represented by the

BR formula: t, 8 oo . . v ~ : JN %, 0 COOH } ...{1VA)

In the above formmlae broken line attachment of a substituent to the bicyclic nucleus : indicates that the substituent is below the plane of r the bicyclic nucleus. Such a substituent is said to be in the A-configuration. Conversely, solid line attachment of a substituent to the bicyclic nucleus 16 indicates that the substituent is attached above the plane of the nm~levas. This latter configuration is referred to an tha Poeonfiguretion. 8 | yer -

FREES )

BAD ORIGINAL 7

L = 3

0

Alao herein reference is made to certain derivatives of cephalosporanic acid, which has the formula:

H

Z ec “LIN 7 | 1 53

LW) 3 -0-

RANG \ , 7 i Cll =0-=CHy

COOH

5 in formula V, Lhe hydrogen ab C 6 is below the plane of the bicyclic nucleus. The derived Lerms desaceloxycephalopporanic acid and 3 desacetoxy methy lcephalonporanic acid are uned to refer to the . formulae VI and Vil, respectively.

H

2 9 H 0 3 n } ,

COOH nOOH

V1 VT

As used herein, 4 ecrotonolactonyl and ¥- “ butyrolacton 14 yl refer to formulae VIII and IX, respectively. The wavy lines are intended to denote ] ’ each of the two epimers and mixtures thereof. | \ \ ¢] ~ 0 0 , - VITI TX . 9 \ 0

BAD ORIGINAL , \

i

When R is an ester-forming residue readily hydrolyzable in vivo in a compound of formula IA, it is a grouping which is notionally derived from an alcohol of the formula R-OH, such that the group - 85 COOR in such a compound of formula IA represents an ester grouping. Moreover, R is of such a nature that the group -COOR is readily cleaved in vivo to liberate a free carboxy group (COOH). That is to say, R is a group of the type that when a compound of formula IA, wherein R is an ester-forming residue readily hydrolyzed in viva, is exposed to mammalian blood or tissue, the compound of formula IA, wherein

R is hydrogen, is readily produced. The groups represented by R are well-known in the penicillin art. In most instances they improve the absorption characteristics of the penicillin compound.

Additionally, the group R should be of such a nature that it imparts pharmaceutically-acceptable properties to a compound of formula IA, and it . 20 liberates pharmaceutically-acceptable fragments when ’ cleaved in vivo.

Typical groups for R are 3-phthalidyl, 4- crotonolactonyl, §-butyrolacton-4-yl and groups of ; the formulae: 2p

= me e— ss wt" rR? © RS © -c-0-C-RP and -c-0-C-0-RP r4 la

X XI wherein each of R3 and R? is hydrogen, methyl or ethyl, and RS is alkyl having from 1 to 8 carbon atoms. However, preferred groups for R are alkanoyloxymethyl having from 3 to 8 carbon atoms, 1-(alkanoyloxy)ethyl having from 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)ethyl having from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having . from 8 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1-methyl-1~

Crewe Sonat + + (alkoxycarbonyloxy)ethyl having from 51to:8. carbon cet atoms, 3-phthalidyl, 4-crotonolactonyl and ¥- butyrolacton-4-yl.

The compound of Formula I above may be 16 prepared by oxidation of either of the compounds of

Formula II or III above. A wide variety of oxidants known in the art for the oxidation of sulfoxides to sulfones can be used for thir process. However, : particularly convenient reagents are metal . 20 permanganates, such as the alkali metal permanganates and the alkaline earth metal permanganates, and organic peroxy acide, such as ] organic peroxycarboxylic acids. Convenient yl individual reagents are sodium permanganate, potassium permanganate, 3d-chloroperbenzoic acid and peracetic acid.

When a compound of the Formula II or III, is oxidized to the corresponding compound of the

Formuia | using a metal permanganate, the reaction is usunaily carried out by reacting the compound of the Formula 11 oe Vid with from 8 5 np 5 molar cquivalenty of the permanganntos. and preferably about 1 molar cauivalent of the rermannanate, in an appropriate aclvenp ryatem. An aprropriate polvent system in one that doen nnt advoreely Interact with aither the starting materiata op the rroduct, and water io commonly weed. Tf deedired, n co-rolvent 5 which is miscible with water but will not interact with the pormancanate, such ap tertrobydrofuran, can be added. fhe reaction ia nermal ly carried out at a ” temperature within the range of from 000 to 56°C. , and preferably at abont 80°C. Ax About f°, the reaction is normally Auhatant inl lv comploke within a ] short period, e.g. within ona hour Although the reaction wav be carried ont gender nebryal, basic or acid conditions, it ia preferable +. oyerato under ’ substantially nentra) conditions in nrdor Lg avoid ) 25 decomposition of the B-tactam ring aviation of the compound of the Formnla 1 Undead it 449 aftan advantagenua tg buffer tha pH wf *he reackion medium in the vicinity of neutrality. The product is i . 4

Le by oD ORIGINAL 9

NAR

. :

ee ee (10 . ye recovered by conventional techniques. Any excess permanganate is usually decomposed using sodium bisulfite, and then if the product is out of solution, it is recovered by filtration. It is separated from managanene dioxide by extracting it into an arganic solvent and romoving the solvent hy evaporation. Alternatively, if the product ia not out of solution at the end of the reaction 1t is isolated by the umal procedure of solvent extraction.

When a compound of the Formula 11 ot IIT is oxidized to the corresponding compound of the

B Formula 1, using an organic peroxy acid, e.g., a peroxycarboxylic acid, the reaction is usually 156 carried out by treating the compound of the Formula

II or II1 with from | to 4 molar equivalents, and preferably about 1.2 equivalents of the oxidant in a reaction- inert organic solvent. Typical solvents are chlorinated hydrocarbons, such as dichloromethane, chloroform and 1,2-dichlorvethane; and ethers, ruch as diethyl ether, tetrahydrofuran and 1,2 -dimethoxyethane. The reaction is normally carried out at a temperature within the range of from - 20° to 509C., and preferably at about 25°C.

At about 257C. reaction times of 2 to 16 hours are commonly used. The product is normally isolated by 2

Le removal of the solvent by evaporation in vacug. The ’ product may be purified by conventional methods, well-known in the art.

When oxidizing a compound of the Formula II ’ Bb or I1I1 to a compound of the Formula I using an ’ organic peroxy acid, it is sometimes advantageous to add a catalyst such as a manganese malt, e.g. manganic acetylacetonate.

The compound of the Formula I, wherein R! 1e is hydrogen, also may be prepared by removal of the protecting group R! from a compound of the Formula

I, wherein R! is a penicillin carboxy-protecting group. In this context, rl may be any carboxy- protecting group conventionally used in the penicillin art to protect carboxy groups at the 8 position. The identity of the carboxy-protecting group is not critical. The only requirements for the carboxy-protecting group rl are that: (1) it . must be stable during oxidation of the compound of

Formula II or III; and (ii) it must be removable . from the compound of Formula I, using conditions under which the p-1actam remains substantially " intact. Typical examples are the tetrahydropyranyl - group, the benzyl group, substituted benzyl groups (e.g. 4-nitrobenzyl), the benzylhydryl group, the 2,2,2-trichloroethyl group, the t-butyl group and the phenacyl group. The penicillin carboxy-

b . - \ . . } . > ¢! ~lr protecting grour is removed in conventional manner, having due resard for the lability of the p- factam ring system. in ike manner, compounds of the Tormula 1, i vie roin i ican previousiyv defined, may be propared by oxidation of a compound of the formula : 4 Sp 3 of N oy, coorl So. (IV) vherein it! is ans proviously defined. This is carried out in exacily the same manner as described hereinbefore for oxidation of a compound of the ) Formula II or 111, except that twice as much oxidant ‘ is usually used.

Compounds of the Formula IA or I, wherein R or rl is an gstev-{forming residue readily hydrolyzable in vive, may be prepared directly from the compound of Formula iA or 1, wherein B or Rr! is hydrogen, by estarification. The specific mathod chosen will depsnd naturally uvpon the precise structure of tho ester-forming residue, bub an appropriates mathod will be readily seleched by one skilled in the art. In the case wherein R or rl is 3-phthalidyl, 4-crotonolactonyl, butyrolacton-4- vl or a group of the Formula X or Xl, wherein R3, Rr : 15 . , \— A

GAD ORIGINAL &

Co n (re and RS are as defined previously, they may be prepared by alkylation of the compound of Formula IA - or I, wherein R or rl is hydrogen, with a 3- phthalidyl halide, a 4-crotonolactonyl halide, a y- butyrolacton-4-yl halide or a compound of the formula rR? © R3 0 enn ev ond

Q-C-0-C-R or Q-C-0-C-0O-R : r r4

X11 XIII wherein Q is halo, and RJ, RY and R® are ae previously defined. The terms “halide” and “halo” are intended to mean derivatives of chlorine, bromine . and iodine. The reaction is conveniently carried out by dissolving a salt of the compound of Formula

IA or I, wherein rl is hydrogen, in a suitable, polar, organic solvent, such as N,N- 156 dimethyl formamide and then adding about one molar equivalent of the halide. When the reaction has proceeded substantially to completion, the product = is imolated by standard techniques. It is often : sufficient simply to dilute the reaction medium with an excess of water, and then extract the product into a water-immiscible organic solvent and then recover the same by solvent evaporation. Salts of the starting material which are commonly used are

Co ——————————————————————————————————————————— 1 y¥ alkali metal anita. ancl of atium and potas in anit and tert Vary Amine cp bat, auch aw

Lriethy amine, t Shy piper ioe. M,N dimethy land tine ant H med iy Tied phot ine antts. The bh reaction 1p run abo {ampere in Lhe range from - about 0 to tone avd ner bby al phot aL The tenp th of ime pnoeded Bo ponch complation varies aecording fo AQ ynarieoty at factors. anech as the concent ration nf Lhe pire tant nl the pee! yvaity of pe the ponent Wher whe cna tedor ind the halo campotvd, ihe jortide pret a pater thee ho tomo. which in turn ric tn { Aster han the Chloride. Tn . fact. il y3 aqomat ines Ad ap Lape onit. when ati bicinm a chloro compre poy oul uw Voy nme mo tar equivalent iH of im atkali peskin podide. This han the at fect of ppoesdingg vy Phe ro cobiden wih tall regard tor thao fore {chore pepe tbe” Limes of from Atuntd to 24 howr:s ee pommont by STR penicil yanic acid lo oxide, the componnd at

Formula 11 wherein r! ia hydrogen, can be prepared by debrominat son of 6.6 Aipremepeniciblanie acid Td oxide. The dehrominat fon can her carried out. uping @ - convent. ion! hydrogen Lysis technique. Thun, - golul ion nl Hh dibromoponiei hanie acid ct oxide 14 ih at ivved ov shaken wnder av at mosphere of hydrogen, or hydro mi eed with an vet Ai nent pach ars nilrea ne tn Cipro IN TE ERIS AL ni a catalytic amend oto JY adem ef horinm core Lomita antalynt. \ 3 ‘BAD ORIGINAL J

Ase --

} . AO

Convenient solvents for this debromination are lower-alkanols, such as methanol ; ethers, such as tetrahydrofuran and dioxan; low molecular weight esters, such as ethyl acetate and butyl acetate; water; and mixtures of these solvents. However, it ie usual to choose conditions under which the dibromo compound is soluble. The hydrogenolysis is usually carried out at room temperature and at a pressure from about atmospheric pressurs to about 50 1e P.8.i. The catalyst is usually present in an amount from about 10 percent by welght based on the dibromo compound, up to an amount equal in weight to the dibromo compound, although larger amounts can be

Co used. The reaction commonly takes about one hour, after which the compound of the Formula 11, wherein

CTT pt is hydrogen, is recovered simply by £1ltration oo followed by removal of the solvent in vacuo. 6,6-Dibromopenicillanic acid lo-oxide is prepared by oxidation of 6,6-dibromopenicillanic acid with 1 equivalent of 3-chloroperbenzoic acid in tetrahydrofuran at 0-25°C. for ca. 1 hour, according to the procedure of Harrison et al. Journal of the

Chemical Society (London) Perkin 1, 1772 (1978). 6,6-Dibromopenicillanic acid is prepared by the method of Clayton, Journal of the Chemical Society (London), (C) 2123 (1969).

gut

Penicillanic acid 1p-oxide, the compound of the Formula III wherein rl is hydrogen, can be prepared by controlled oxidation of penicillanic acid. Thus, it can be prepared by reacting penicillanic acid with one molar equivalent of 3- chloroperbenzoic acid in an inert solvent at about 20°C. for about one hour. Typical solvents are chlorinated hydrocarbons, such as chloroform and ‘ dichloromethane; ethers, such as diethyl ether and 19 tetrahydrofuran; and low molecular weight esters such as ethyl acetate and butyl acetate. The product is recovered by conventional techniques.

A compound of the Formula II or III, wherein rl is an ester-forming residue readily hydrolyzable in vivo, may be prepared directly from the compound of Formula II or III, wherein rl is hydrogen, by esterification, using standard procedures. When r} is 3-phthalidyl, 4- crotonolactonyl, ¥ ~butyrolacton-4-yl or a group of the Formula X or XI, wherein rR3, r3 and RO are as defined previously, the compound may be prepared by alkylation of the appropriate compound of the

Formula II or 111, wherein rl is hydrogen, with a 3- phthalidyl halide, 4-crotonolactonyl halide, a Y - butyrolacton-4-yl halide, or a compound of the

Formula XII or XIII. The reaction is carried out in exactly the same manner as described previously for esterification of penicillanic acid 1,1-dioxide with

3 gil a 3-phthalidyl halide, a 4--crotonolactonyl halide, a

J-butyrolacton-4-yl halide, or a compound of the

Formula XII or XIII. } Alternatively, the compound of the Formula .

II, wherein rl is an ester-forming residue readily hydrolyzable in vivo, can be prepared by oxidation of the appropriate ester of 8,6-dibromopenicillanic ‘ acid, followed by debromination. The ester of 6,6- dibromopenicillanic acid is prepared from 8,6 190 dibromopenicillanic acid by standard methods. The . oxidation is carried out, for example, by oxidation - with one molar equivalent of 3-chloroperbenzolc acid, as described previously for the oxidation of . 8,8-dibromopenicillanic acid to 6,6- 16 dibromopenicillanic acid lc-oxide; and the debromination is carripd out as described previously for the debromination of 6,8-dibromopenicillanic acld l1d-oxide.

In like manner, the compound of Formula 111, wherein Rr! is an ester-forming residue readily hydrolyzable in vivo can be prepared by oxidation of the appropriate ester of penicillanic acid. The ’ latter compound is readily prepared by eaterification of penicillanic acid using standard methods. The oxidation is carried out, for example,

—r— — ptt? by oxidation with one molar equivalent of 3- chloroperbonsoic ncid, an deaoribed proviously acid 1f-oxide.

The compound of the Formula 117, wherein Rr! bh is a carboxy protecting gronp may be prepared in one of two ways. 1t may be prepared by attaching A carboxy protecling group Lo penicillantie acid 14 oxide. Alternatively, it may be prepared by: (a) attaching a carboxy protecting group tao 6,6 dibromopenicillanic acid; (b) oxidizing the protected 6,06 dibromopenicillanic acid to a protected 6,6 dibromopenici Janie acid ld-oxide using | molar cquivalent of 3 chloroperbenzoic acid; and (¢) debrominating the protected 6,6

De . co . Cw EK eas . , , ee Nea oe cL . dibromopenicillanie acid oxide hy hydrogenolysis.

The compound of the Formula 111, wherein rR} . is a carboxy protecting group may be prepared by attaching a protecting romp Lo penicillanic acid 14- oxide. Alternatively, it may be prepared by: (a) attaching a carboxy protecting group to penicillanic } . acid; and (b) oxidizing the protected penicillanic acid using | molar equivalenl of 3-chloroperbenzoic acid as previounly described

The compounds of Formulas 1, IT and I11, wherein r! ig hydrogen, are acidic and will form salts with bagic arent. Such salta are within Lhe 2

TE WY

BAD ORIGINAL J

: g1d

Q¥ scope of this invention. These salts can be prepared by standard techniques, such as contacting the acidic and basic componenbs, usually in a 1:1 molar ration, in an aqueous, non-aqueous or partially aqueous medium, as appropriate. ‘The salts are then recovered by filtration, by precipitation with a nmm-solvent followed by Filtration, by evaporation of the solvent, or in the case of aqueous solutions, by lyophilization, as appropriate. Bases which are suitably employed in galbt formation may be organic or inorganic, and suitable examples are ammonia, organic aminen, alkali metal hydroxides, carbonates, bicarbonate, hydrides and alkoxides, and alkaline earth metal hydroxides, carbonates, hydrides and ‘alkoxides. Representative examples of such bases are primary amines, such as n-propylamine, n- butylamine, aniline, cyclohexylamine, benzylamine : and octylamine; necondary amines, such as diethylamine, morpholine, pyrrolidine and piperidine; tertiary amines, such as triethylamine,

N-ethylpiperidine, N methylmorpholine and 1,5- diazobicyclo [4.3.01non 9H ene; hydroxides, such as ‘ sodium hydroxide, potassium hydroxide, ammonium hydroxide and barium hydroxide; alkoxides, such an ; 25 sodium ethoxide and potassium ethoxide; hydrides, ‘ such as calcium hydride and sodium hydride; . cabonaten, such as potassiuvom carbonate and sodium { } 27

BAD ORIGINAL 2 ’

L

—— rt ——r eee Aer ee —— ee (vo ys carbonate; bicarbonatea. such as sodium bicarbonate and potassinm hicarbonake; and alkali metal salta of long chain fatty acida, such as sodium 2- : cthy lhexinoate.

I'referred saltae of the compounds of

Formulae 1, 11 and UIT ave sodium, potassium and : triethylamine salli,

Ann indicated hereinbefore, the compounds of

Formula IA, awl Lhe phaamacontical ly acceptable galt thereof are antibacterial agents of medium potency. The ip vilkro activity of the compound of ‘ the Formila 1A, wherein R in hydrogen, may be denonatrated by meararing its minimom inhibitory concentrations (MIC =) in mop, ml apainat a variety © : I. . Aa te Ps qt? = . : SoA « Oy hb want My Ge pe Rg res . of microorganiems. The procedure which ig followed is the one recommended by the International

Collaborative Study on Antibiotic Sensitivity

Testing (Friccson and Sherris, Acta.Pathologica et

Microbiologia Ocandinav, Supp. 217, Sections A and

B: 1-90 {19791), and employs hrain heart infusion (BHI) agar and the inocula replicating device.

Overnight growth tubes are diluted 100 fold for use as the ntandard inoculum (20,000 10,000 cells in ‘ approximately 9.902 ml. are placed on the agar ] sur face; 29 ml. of Wil apar/dish). Twelve 2 fold dilutions of the tent compound are employed, with init al concentration of the test drug being 200 oN . Pr

BAD OHIGINAL i

¢ 10

Le meg. /ml. Single colonies are disregarded when reading pintes alter 18 hrs. at 379°C. The susceptibility (MIC) of the test orgnaism is accepted as the lowent concentration of compound capable of producing complete inhibition of growth as judged Ly the naked eye. MIC values for . penicillanic acid 1,1 dioxide against several microorganisms are shown in Table 1. , . 24 ‘BAD ORIGINAL gl a 4 dd ee - _

Lv gi

TABLE 1

In Vitro Antibacterial Activity of —Penicillanic Acid l.l1-Dioxide

Microorganism MIC (weg./ml.)

Staphylococcus awreurn ige ‘

Streptococcus iaecalis >200

Streptococous progenen 190 }

Escherichia coli. 50

I'seudomonas aeruginosa 200

Klebsiella pneumoniae Ho

Proteus mirabilis 100

Proteus wargtani. 100

Salmonella typhimurium 50 , Pasteurella multocida He

Co 7 1B Serratia marcescens . 100

Enterobacter aerogenes 25

Enterobacter clocae 100 ”

Citrobacter freundii 50

Providencia 100

Staphylococcus epidermis 200

Pseudomonas putida >200

Hemophilus influenzae > 50

Neisseria gonorrhoeas | 0.312 26 > 'BAD ORIGINAL ol

Le

: The compounds of tha Formula IA, and salts thereof are active on antibacterial ag-nis in viva.

In determining such artivity, acute experimental infections are produced in mice by the intraperitoneal inocnlaklon of the mlee with a . standardized culture of the test orgavlrm suspended in 5 percent hos gastric mucin. Tnfectjon severity is standardized ro that the mice receive one to ten : times the Doe dose of the organism (IDgpp: the minimum inoculum of organism required to coneistently kill 199 percent of the infected, non- treated control mice). The test compound is administered to the infected mice using a miltiple dosage regimen. Av the end of the test, the ’ 15 activity of a compound ls assessed by counting the : number of survivors among the treated animals and expressing the activity of a compound as the percentage of animals which survive.

The in vitro antibacterial activity of the . 20 compound of the Formula IA wherein R is hydrogen : makes it useful as an industrial antimicrobial, for example in water treatment, slime control, paint > . preservation and wood preservation. as well as for topical application as a disinfectant. In the case of use of this compound for topical application, it is often convenient to admix the active ingredient with a non-toxic carrier, such as vegetable or mineral oil or an emollient cream. Similarly, it f oR g | i

BAD ORIGINAL 9 ee e—_—

TV can he dissolved or dispersed in liquid diluents or solvents such as water, alkanols, glycols or mixtures thereof. In most instances it is appropriate to employ concentrations of the active ingredient ol irom about 9.1 percent to about 10 percent Ly woighti, basad on cetal composition.

The in vive activity of the compounds of :

Formula lA, aud salts thercof makes them suitable for thu vonlrul of bacterial infections In masmalz, including man, by bolh tho gral and parenteral miles of adminirlrabion. ‘The compound: will find use in the :ontrol of infections caused by munceptible bacterla in human subjects, ©.g- infections cavsed by strains ol Neisseria genorrhoesa. 16 When considering therapeutic use of a compound ol the Formula lA, or a salt thoreof, in a mammal, parhicularly man, the compound may be administered alone, or it can be mixed with pharmaceutically acceptable carriers or diluents.

They may be adninistered orally or parenterally, j.e. intramuscularly, subscutancouoly or intraperitoneally. The carrior or diluent is chosen on the basis of the intonded modo of administration.

For example, when considering the oral mode of 26 administration, an antibacterial penam compound of ’ this invention may be used in the form of tablets, capsules, lozenges, troches, powders, syrups, pi

BAD OHiGINAL 9

L gq! 'V i elixirs, aqueous solutions or suspensions, in oo accordance with standard pharasceuntical practice.

The prouportion=) ratio of active ingredient tn carrier will naturally der=nd on the chemical natura, roluvbility and stobility of the artive ingredient, n= well a= the do-age contemplated. . However, rharmacontical compositions containing an antibacterial pont of Fhe Formmla TA will likely contain from 20¥ +n 95% nf active ingredient. In the come of tablets for oral nse, carrices which are commonly u=ed luclinde lactose, sodium citrate and salta of phosphoric acid. Varioun disintegrants such aa starch, and lubricating agontn, ouch as magnesium stearate, sodium lauryl sulfate and tale, i5 are commonly used in tabletn: For oral administration in capsule form, nzeful diluents are lactone and hirh molecular weight rolyethylene : glycols. When aqueous suaperelions ara required for oral use, the active ingredient. is combined with emulsifying and suapanding agenta. If desired, : certain sweetening and/or flavoring agents can be added. For parenteral administration, which includos intramuscular, intraperitoneal, ’ subcutaneoua and intravenous use, sterile solutions - 25 of the acbive ingredient are usually prepared, and the pil of the solutions are suitably adjusted and ’ Bh 28 ow

BAD ORIGINAL J}

a buffered. For intravenous use, the total concentration of solutes should be controlled to render the preparation isotonic.

As indicated earlier, the antibacterial . 5 agents of this invention are of wie 1a human subjects against susceptiltile orgaaisma. The prescribing physician will vlitlmately determine the appropriate dose lor a given human subject, and this can be expected to vary according to the age, / 10 welght, and response of the individual patient, as well as the pa'mre and Lhe sciceity of the patients symptoms. The compounds of Lhis fovemb ion will normally be used orajly ab docagas ia Lhe range from to 200 mg. per kilogram of body wealght per day, oe 15 | and parenterally at dosages fcom 10 to 400 mg. per Co ) kilogram of body weighl per day. These figures are illustrative only, however, aud in rome cases it may / be necessary Lu use dosages outside Lhese limits.

Additionally the compounds of the Formula

IA and malts thereof are potent inhibitors of microbial p-1actamanes, and they increase the antibacterial effectiveness of p-1actam antibiotics (renicillinng and cephalosporins) sgeinst many microorganisms, vacticularly those which produce a p- lactamase. The mammer In which the comrounds of the

Formula {A increase Lhe clfectiven:== of a p-lactam antibiotic may be appreciated by referomne to iJ : oo

BAD ORMGINAL gy qv © experiments in which the MIC of a given antibiotic alone, and a compound of the Formule IA alone, are measured. Thcue MICU 2 are thon compared with the

MIC values obtained wilh a cembive tion of the given b antibiotic and the compound of the Furmula IA. When the antibaclerial potency of the combination is significantly greater than would have Leen predicted . from the poulunvies of tho individual compounds, this is considered Lo constitute enhancemsnl of activity.

The MTC values of combinations ars measured uning the method described by Barry and Sabath in "Manual ‘of Clinical Microbiology”, edited by Lenette,

Spaulding and I'rauni, 2nd edition 1974, American

Bociety for Microbiology. } Results nf expariments illustrating that penictllante acid t,1-dioxide enhances the effectivenanr of ampicillin Are raported in Table

IT. From Table 11, it can he meen that against 19 ampicillin-reristant, strains of Staphylococcus aurays, the moda MIC of ampicillin, and of penicillaniec acid 1.1-dioxide. is 200 mcg. /ml.

However, tha mode MIC's of ampicillin and i penicillanic acid 1,1-diloxide in combination are 1.568 and 3 12 meog/nml . vespeclively. Looked at another way, thin means that whereas ampicillin alone has a mode MIC of 209 mca. /ml. against the 19 strains ot Sharhelococous anrens, its mode MIC is v : raduced to L. BG mea. /m). in the precencs of 3.12 . 39

BAD CrulaiiaL 9)

. . | (0

Ju mcg./ml. of penicillanic acid 1,1-dioxide. The other enlvico in Table IT ghow enhancement of the antibaclurial effectiveness of ~mpici!lin apgainat 26 ) ampicillin resintant trainee of Haenaphi tue : intluencae, 10 ampicillin resistant strains of

Rlebaiella pucumonine, and 1% straing of the anerobe

Dacterides fragilis. Tables ITT, 1V and V show enhancement. of Lhe antibacterial potency of beunsylpenicillin (yeniciliin G) carbenicillin ( - 19 carboxybenrylpenicillin) and cefazolin, respectively, against strains ot §. aureus, H. influenzae, K. penumoplace and fBacterpidea fragilis.

Te tel Cee me . we CE eam ey oo , 31 cere : — TT)

BAD Cin :

N

(0 vt . ) Oo» ut ® 6 . . g 2 ; g . ot o . 0 * CL ’ * - } g ; LF a : hb Bb 8 © : 8 b : Eo =

B+ B 0 be , pox ? . "i E

S g $ aa | « . : : we = 3

Cc | 5 ¥ a . LF 0

Hh

S wd dg °% >

Vv fb ed fab 3 ER $8 0 .

Br fe & [ CY 8 8

Ad ) 0 + g s an xX ct o 032 [= oo con El

J oO oo Oo = 5 > XE - of 0 ® ba

A Se rg Wr Fh . > - | ER . o eo © ow ow | fg 3s B® BB = BE od 1 a ® 8 : 32

A

. JD

Ww : on 7

K BE rA | x

EE EE od

Hook on z 6 E = Ro 5 lB a Q ®

Bo. op = nor . b & E&OE 5 2 +g +

J. | ® : 5 0 a 4 = ; ne .

Eg g Ee | a ) iV : 8] 2 i : -

J TTT TTY MATRA 2 =n fas 2F : i — ttm | Ho b mos rm Eo H

Ne g 7 3 —— rr ——— ey py —— = . } oo X

HQ P on § o. ’ 0 % i a, , B® 3 3 B32 bs 3 8 g ’ a.Q *i, Yeon ie . . x en een Lo B® oa Bu : [ER To | + ceil MI — og p 33 5 i 3 we ee eT TY $e

EI i pox ® 5a ~ " 590 ™ ox <B>

BE

= Bb) . «0 22 8 2 - i Ey - = | co 2 ’ | : 3 n b & 8

Sr een 5

Re 5

I.

Pow ow X &

Low o.oo |B LEER 3 / . ’ ” 2 ‘ ’ ~~ | or — 0 ’ r.. ts . we : / 9 A 5 1 > 3 tg : / 5 gra or oes ! ——— oo - 9g i rt fo dy . — £3 ® =D | a be

Ct ogres Pon 5 i & mh | 5 .

ERE: [I 0 } A

Cri NY os

BAD UHIGINAL

7° feb = on pre :

EBB 1

I Boe Ey 3

B = 8

BE FE | : } “ 8 be Y q

CEs Ee | : & gE § E | : £, be. 5 n co

Eo& BR O§ 8 { nz Bs i 0 a

I a = 7 FS - B Ju eo . § i ee " ~ 3 5 oo x igs | LS . o A o ro | : g | v 2 Yo peo » tw =e 30 3 x | 4 : : + : 8 ®»oX 8 252 S

AN > in: . a bg «0 mow 3 T T ® © 8 > Le o 3 : 8 3 2 - | 8 | § : : hd . . ! 0 0 . 38% : o 5 BB |E BE pega of gc =u ’ . z ) 0 s | fs > a [u oH mo GW om ~ | og “hy » te 5E mo ® 5 a i | gs - - a. bo . & 5 ® 5 PR . . I - ¢ re 3 BAD ORIGINAL 9

_— — a ® on - x : fn : g : . ct : Q a] - g §F ¥8 3 = 58

E o 0 >

Ne go 8 gE 8B o In IX a ’ [al fb o g od 3 o> > ’ eB " $ nN ne HR 2 g 3 o g 5 8 b & o 8 . 8 . a Q et ooo | § | BEE ¥

B 8B BF © £ box

E| 2.8 : pote w » go vg = 8 i» § - | Ed o 5 oy Ih | EE 5 08 ¥ | EE { » E ; - : j=) o

I - fo .

A : 35

So {

oo | pA

The compounds of the Formula IA, and salts thereof enhance the antibacterial effectiveness of p-lactam antibiotics in vivo. This is, they lower the amount of the antibiotic which is needed to protect mice against an otherwise lethal inoculum of certain B-lactamase producing bacteria.

The ability of the compounds of the Formula

IA, and salts thereof to enhance the effectiveness of a B-lactam antibiotic against f-lactamase- producing bacterial makes them valuable for co- administration with B-1actam antibiotics in the treatment of bacterial infections in mammals, particularly man. In the treatment of a bacterial infection, the said compound of the Formila IA, can : . be co-mingled with the p-lactam antibiotic, and the : ’ two agents thereby administered simultanecusly.

Alternatively, the said compound of the Formula IA, can be administered as a separate agent during a : course of treatment with a f-lactan antibiotic. In some instances it will be advantageous to pre—dose : ~ the subject with the compound of the Formmtla IA, # before initiating treatment with a p-lactam } antibiotic. RE : n : \ ) When suing penicillanic acid 1,1-dioxide or an ester thereof readily hydrolyzable in vivo to enhance the effectiveness of f-lactam antibiotic, it 38 Co

— “1 - ims administered preferably in formulation with standard pharmaceutical carriers or diluents. The i methods of formulation discussed earlier for use of penicillanic acid 1,1-dioxide or an ester thereof b readily hydrolyzable in vivo as a single-entity antibacterial agent can be used when co- administration with another b-lactam antibiotic is intended. A pharmaceutical compogition comprising a pharmaceutically-acceptable carrier, a p-lactam antibiotic and penicillanic acid 1,1-dioxide or a ’ readily hydrolyzable ester thereof or a salt thereof will normally contain from about 6 to about 80 percent of the pharmaceutically acceptable carrier by weight.

Cl to . 15 ne When using penicilianic acid 1,irdioxide or .. © . an ester thereof readily hydrolyzable in vivo in : combination with another p-lactam antibiotic, the sulfone can be administered orally or parenterally, i.e. intramuscularly, pubcutaneously or intraperitoneally. Although the prescribing physician will ultimately decide the dosage to be used in a human subject, the ratio of the dally dosages of the penicillanic acid 1,1-dioxide or ester or salt thereof and the p-1actam antibiotic 26 will normally be in the range from about 1:8 to 3:1.

Additionally, when using penicillanic acid 1,1 dioxide or an ester or salt thereof in combination ot with another p Factam antibiotic, the daily oral dosage of each component will normally be in the range from about 10 to about 200 mg. per kilogram of . body weight. and the daily parenteral dosage of each component will normally be about 10 to about: 400 mg. per kilogram of body weight. These figures are illustrative only, however, and in some cases it may be necessary t.o use dosages outside these limite. ’ "Typical p-1actam antibiotics with which penicillaniec acid \,1-dloxide, or ester: or salts thereof may be co-administered are: 6-(2-phenylacetamido)penicillianic acid, 6-(2)- phenoxyacetamido)penicillanie acid, . . 6-(2--phenylpropionamido)penicillanic acid, 6-(D-2-amino -2- phenylacetamido)penicillanic acid, 6-(D-2-amino-2-[4 -hydroxypheny 1 Jacetamido )- . penicillanic acid, 6-(D-2-amino-2 11,4 eyclohoxadienylJacetamido)- } renicillanic acid, 6-(1-aminocyelohevanecarboxamido)penicillanic acid, 6-(2-carboxy -2- phenylacetamido) pencil lanic acid, 6—(2-carboxy--2 [J thienyl lacetamido)penicillanic ’ . anid, 6-(D-2-{4-ethylpiparazin 2,3 dione | -carboxamido }- 2 -pony tacatamido)penicil lanie acid, 6-(R-2-T4 hydroxy t,5-naphthyridine-3-carboxamido]- . < 2 -phenylacetamido)penicil lanice acid, an oo

CL

SE BAD ORIGINAL Pp} . " ‘

—————— ee —————————— + _ 6 (D 2 snlfn 2 phonylacetamido)penicitlanie acid, - 6 (D2 aulioaming © phenytacetamido)penic tllanic . acid,

G-L ¥ limidarolidin 2 one 1 carboxamidol-2-

Hh phenviacetomidoipenicillanic acid,

G6 (1 [3 mathybmibionyiimidasobadin 0 one carboxamido] on pineny lace tamido )penici l tanice acid, 6 (1 heyvahydro 1H azepin | yl Imethyleneamino) i0 ponicil lanie acid, acetoxymeihyl 6 (2 pheny lacetamido)penicillanate, acetoxymethy!l 6 () 2 amino 2 phenylacetamido) ponict i banal.e, . acetoxymethy tL 6 (D 2 amino 2 [4 hydroxyphenyl 1 oo acetamido)penicillanate, Cees Le pivaloyloxymethyl 6 (2 phenylacetamido)penici Llanate, pivaloyloxymethy! 6 (1) 22 amino 2 pheny lacetamido) penicil lanate, ‘ pivatloyloxymethy! 6 (D2 amino-2 [4 hvdroxypheny ld 29 ace tamido)penicil lanate, 1--{ethoxycarbonyloxylethyt 6 (2 phenylacetam ido) penicillannte, 1 (ethoxycearvbonyloxy)ethyl 6 (D2 amino 2 phenyl : acetamido)penicil lanate, 25H 1-(ethoxycarbony loxy)ethyl 6 (D2 amino 2 141 hydroxyphenyl lacetamido)penicillanate,

A-phthalidyl 6 (2 phenylocetamide)penicillanate,

I)

BAD ORIGINAL 9

. 0

A

3-phthalidyl 8-(D-2-amino-2-phenylacetamido)- penicillanate, 3-phthalidyl 6 (D-2-amino-2-[4-hydroxyphenyl ]- to acetamido)penicillanate, b 6- ( 2-phenoxycarbonyl-2-phenylacetamido)penicillanic . acid, : 6-(2-tolyloxycarbonyl-2-phenylacetamido)penicillanic acid, 8-(2- [5-indanyloxycarbonyl]-2-phenylacetamido)- penicillanic acid, 6-(2-phenoxycarbonyl--2-[3-thienylJacetamido)- penicillanic acid, 8- (2-tolyloxycarbonyl-2-[3-thienyllacetamido)- penicillanic acid, 8- (2- [5-indanyloxycarbonyl-2-[3--thienylJacetamido)- 16 penicillanic acid, 6~-(2,2-dimethyl-5-oxo-4-phenyl--1-imidazolidinyl)- } penicillanic acid, 7-(2-[2-thienyl lacetamido)cephalosporanic acid, 7-(2-[1-tetrazolyllacetamido-3-(2-[56-methyl-1,3,4- thiadiazolyllthiomethyl)--8- desacetoxymethylcephalosporanic acid, } 7-(D-2-amino-2-phenylacetamido )desacetoxy— cephalosporanic acid, ) 7-o-methoxy-7-(2-[2-thienyl]acetamido)-3- carbamoyloxymethyl-3-desacetoxymethyl—- cephalosporanic acid, ’ 7-(2-cyanoacetamido)cephalosporanic acid, _ ae ;

et ee ee eet eet et tee ——————————————— ee _________ oo | gtd ov - 7—(D hydroxy 2 pheaylacetamido) 3 (H Ii mathy!tbebrazolyl ILhiomethylt) 3 desacetoxymethy leophaloaporanic acid,

T—(2-- 14 pyridylthio Iacetamida)eephalosporanic acid,

T-(D 2 amino 2 (1,4 evelohexadienyl lacetamido) . cephalosporanic acid,

T--(D 2 imino 2° phony lacebamido)cephaloaporanie acid, and : the pharmaceutically acceptable nalts thereof.

As will be appreciated by one gkilled in the art, some of the above p lactam compounds are effective when administered orally or parenteral ly, while others are effective only when administered by ' oo ' the parenteral route. When penicillanic acid 1,1 | dioxide or an esler or malt thereof is to be used Co ' or “gimultaneounly (i.e. co mingled) with a Pp tactam antibiotic which ig effective only on parenteral use

CL will be required. When the penicillanic acid 1,1 dioxide or cater or salt thereof is to be used agimultaneoualy (co mingled) with a B- lactam antibiotic which is effective orally or parenterally, formulations suitable for either oral or parenteral adminiagtralion may be prepared.

Additionally, it is ponsible to administer formulationn of the penicillanie acid 1,1 dioxide or ester or salt thereof orally, while at the same time administering a further B-1actam antibiotic 41

BAD ORIGINAL 9 i oo aS parenterally; aul it is also possible to administer formulations of the penicillanic acid 1,1-dioxide or eater or salt thereof parenterally, while at the same time administering the further p-lactam antibiotic orally.

The following examples are provided solely for the purpose of further illustration. Infrared : (IR) spectra were measured as potassium bromide discs (KBr dines) or as Nujol mulls, and diagnostic absorption bands are reported in wave numbers (em 1).

Nuclear magnetic resonance spectra (NMR) were measured at 60 MHz for solutions in deuterochloroform (CDClj,), perdeutro dimethyl sulfoxide (DMSO-dg) or deuterium oxide (Dy0), and peak positions are expressed in parts per million (ppm) downfield from tetramethylsilane or sodium 2.92-dimethy1-2- oT silapentane-5-sulfonate. The following : abbreviations for peak shapes are used: a, singlet; ‘ d, doublet; t, triplet; q, quartet; m, multiplet.

- | EEE

EXAMPLE 1

Penicillanic Acid 1.1:Diogida

To a solution of 8.61 gg. (41 mmole) of potassium permanganate in 130 ml. of water and 4.95 ml. of glacial necelble acid, cooled to ca. 5°C., was added a cold (ca. H7C.) volution of 4.50 g. (21 mmole) of the sodiwn sali, of penicillanic acid in 50 : ml. of water. ‘the mixture wos stirred at ca. 59°C. for 20 minutes and then the cooling bath was removecl. Solid sodium bisulfite was sdded until the color of the potassium permanganate had baen discharged, snd then the mixture was flltered. To . co the agueous filtrate was added half its volume of saturated sodium chloride solution, and then the pH }

Cow ’ +16 7 + was adjusted to 1.7. The acidic solwtionwag. ~~ = extracted with athyl acetate. The extracts were dried, and then evaporated in vacua, to give 3.47 g. of the title product. The aqueous mother liquor was saturated with sodium chloride, and further extracted with ethyl acetate. The ethyl acetate . salution was dried and evaporated in vacua, to give a further 0.28 g. of product. The total yield was therefore 3.75 g. (76% yield). The NMR spectrum (DMSO--dg) of the product showed absorptions at 1.40 (8,34), 1.50 (8,31), 3.13 (d of d's, IH, J; = 16Hz,

Jy = 2Hz), 3.63 (d of d"m, 1H, Jy = 16 llz, Jo =

AHz), 4.22 (3, LI). and 65.03 (d of d's, 1H, Jy - 4H=z,

Jog = 2Hz) ppm. 43

I” ORIGINAL dD

9 pé! ’ 1 _ : To a atirred solution of 6.85 g. (24 mmole) of benzyl penicillanate in 75 ml. of ethanol-free : 5 chloroform, under nitrogen, in an ice-bath, was added in two portions, several minutes apart, 4.78 8. of BX pure 3-chloroperbenzoic acid. Stirring ‘was continued for 30 minutes in the ice-bath, and then 46 for minutes without external cooling. The reaction mixture was washed with aguecus alkali (pH 8.6), followed by saturated sodium chloride, and then it was dried and evaporated in vacuo to give 7.86 g. of residue. Examination of this residue showed it to be a 5.5:1 mixture of benzyl ~ penicillanate 1-oxide and benzyl penicillanate 1,1- dioxide.

To a stirred solution of 4.85 g. of the above 5.5:1 sulfoxide-sulfone mixture in 50 ml. of . ethanol-free chloroform, under nitrogen, was added 3.2 g. of 85X pure 3-chloroperbenzoic acid at room . temperature. The reaction mixture was stirred for 2.5 hours, and then it was diluted with ethyl : acetate. The resultant mixture was added to water - at pH 8.9, and then the layers were separated. The : 25 organic phase was washed with water at pH 8.0,

Lo followed by saturated sodium chloride, and then it : was dried using sodium sulfate. Evaporation of the b oo 44 g

—,,,———— «® solvent in vacuo afforded 3.59 g. of the title compound. The NMR spectrum of the product (in

CDClg3) showed absorptions at 1.28 (ss, 8H), 1.58 (s,34), 3.42 (m,2H), 4.37 (s,1H), 4.55 (m,1H), 5.18 b (,2H, J = 12 Hz) and 7.35 (s,bH) ppm.

Co CL Whe bomn, HM VL Id ed a Bre aS Tg SE a eh rm een lane teh ides JER Eh oS

EXAMPLE 3

To a stirred solution of 8.27 g. of benzyl } penicillanate 1,1-dioxide in a mixture of 40 ml. of . 5 methanol and 18 ml. of ethyl acetate was slowly added 106 ml. of water, followed by 12 g. of 5% palladiwm-on-calcium carbonate. The mixture was shaken under an atmosphere of hydrogen, at 652 psi, for 40 minutes, and then it was filtered through 19 supercel (a diatomaceous earth). The filter cake wan washed with methanol, and with agueous methanol,

J and the washings were added to the filtrate. The combined solution was evaporated in vacuo to remove i "the majority of the organic solvents and then the Coe residue was partitioned between ethyl acetate and

Co water at a pH of 2.8. The ethyl acetate layer was . removed and the aqueous phase was further extracted with ethyl acetate. The combined ethyl acetate : solutions were washed with saturated sodium chloride solution, dried naing sodium sulfate and then evaporated in vacua. The residue was slurried in a . 1:2 mixture of ethyl acetate-ether, to give 2.37 g. : of the title product having a melting point of 148- - . 51°C. The ethyl acetate-ether mixture was : 25 evaporated giving a further 2.17 g. of product.

(0

EXAMPLE 4 piyalavlosymethyl Penicillenate 1.1-Dioxide

To 0.615 g. (2.41 mmole) of penicillanic acid 1,1-dioxide in 2 ml. of N,N-dimethylformamide was added ©.215 g. (2.60 mmole) of diisopropylethylamine followed by 9.365 ml. of chloromethyl pivalate. The reaction mixture was stirred at room temperature for 24 hours, and then it was diluted with ethyl acetate and water. The ethyl acetate layer was separated and washed three times with water and once with saturated sodium . chloride solution. The ethyl acetate solution was then dried using anhydrous sodium sulfate, and bo ee .. . evaporated in vacuo to give 0.700 g. of the title 16 product as a 014d, mp 103-4° ie a peck ET me the product (in CNClg) showed absorptions at 1.27 (s, OH), 1.47 (s, SH), 1.62 (8, an), 3.62 (m, 2H), . 4.47 (8, 1H), 4.70 (m, 1H), 5.73 (4d, 1H, J = 6.0 Hz) and 5.98 (d, 1H, J = 6.0 Hz).

EXAMPLE 5

The procedure of Example 4 js repeated, except that the pivaloyloxymethyl chloride used therein is replaced by an equimolar amount of : acetoxymethyl chloride, propionyloxymethyl chloride and hexanoyloxymethyl chloride, respectively, to give: a7

1d ¢l 0) Vv acetoxymethyl penicillanate 1,1-dioxide, proplonyloxymethyl penicillanate 1,1-dioxide and hexanoyloxymethyl penicillanate 1,1-dioxide, respectively.

—-—————

P<

EXAMPLE © 3-Phthalidyl Penicillanate 1.1l-Dioxide

To ©.783 g. (3.36 mmole) of penicillanic acid 1,1-dioxide in 5 ml. of N,N-dimethylformamide was added 8.47 ml. of triethylamine followed by 0.715 g. of 3-bromophthalide. The reaction mixture was stirred for 2 hours at room temperature and then it was diluted with ethyl acetate and water. The pH of the aqueous phase was raised to 7.9 and the layers were separated. The ethyl acetate layer was ‘ washed successively with water and saturated sodium chloride solution, and then it was dried using : sodium sulfate. The ethyl acetate solution was evaporated in vacua leaving the title product as a

Cotte 038 0 white foam. The NMR spectrum of the product (tm.

CDClg3) showed absorptions at 1.47 (a, 611), 3.43 (m, 1H), 4.45 (=, 1H), 4.62 (m, 1H), 7.40 and 7.47 Lo (28°, 1H) and 7.73 (m, 4H) ppm.

When the above procedure is repeated, except that the 3-bromophthalide is replaced by 4- bromocrotonolactone and 4-bromo- {-butyrolactone, : respectively, thie affords: 4-crotonolactonyl penicillanate 1,1-dioxide and ¥-butyrolacton-4-yl penicillanate, respectively.

nS

EXAMPLE 7 1-(Ethoxyvecarbonvloxy)ethyl Penicillanate 1.1-Dioxide

A mixture of @.654 g. of penicillanic acid 1,1-dioxide, 8.42 ml. of triethylamine, 0.412 g. of 1-chloroethyl ethyl carbonate, 0.300 g. of sodium bromide and 3 ml. of N,N-dimethylformamide was stirred at room temperature for 6 daya. It was then worked up by diluting it with ethyl acetate and water, and then the pH was adjusted to B.5. The ethyl acetate layer was separated, washed three times with water, washed once with saturated sodium chloride, and then it was dried using anhydrous sodium sulfate. The ethyl acetate was removed by evaporation ip vacua leaving 9.390 g. of the title product as an oil.

The above product was combined with an approximately equal amount of similar material from . a similar experiment... The combined product was dissolved in chloroform and | ml. of pyridine was added. The mixture was stirred at room temperature overnight and then the chloroform was removed by evaporation in vacuo. The residue was partitioned between ethyl acetate and water at pH . The : separated and dried athyl acetate wan then 26 evaporated in vacuo to give 150 mg. of the title product (yield ga 7X). The IR spectrum (film) of the product showed absorptions at 10056 and 1783 em,

-— . . ne

The NMR spectrum (CDClgy) showed absorptions at 1.43 (m, 12H), 3.47 (m, 2H), 3.9 (a, 2H, J = 7.5 Hz), : | 4.37/m, 1H), 4.63 (m, 1H) and 8.77 (m, 1H) ppm. : EXAMPLE 8 b The procedure of Example 7 is repeated, except that the 1-chloroethylethyl carbonate is replaced by an equimolar amount of the appropriate 1-chloroalkyl alkyl carbonate, I- (alkanoyloxy)ethyl chloride or 1-methyl-1-(alkanoyloxy)ethyl chloride, to produce the following compounds: methoxycarbonyloxymethyl penicillanate 1,1-dioxide, ethoxycarbonyloxymethyl penicillanate 1,1-dioxide, isobutoxycarbonyloxymethyl penicillanate 1,1- a mean ord bt dioxide; pan ete aan ain Tm pe Ra Cee 16 © {_(methoxycarbonyloxy)ethyl penicillanate 1,1- dioxide, 1- (butoxycarbonyloxy ethyl penicillanate 1,1- dioxide, : 1-(acetoxy)ethyl penicillanate 1,1-dioxide, 1-(butyryloxy)ethyl penicillanate 1,1 dioxide, 1-(pivaloyloxy)ethyl penicillanate 1,1-dioxide, oo 1-(hexanoyloxy)ethyl penicillanatae 1,1-dioxide, 1-methyl-1- (acetoxy)ethyl penicillanate 1,1-dioxide and 1-methyl-1-(iscbutyryloxy)ethyl psnicillanate 1,1-dioxide, respectively.

Lest

EXAMPLE 9

The procedure of Example 4 is repeated, except that the chloromethyl rpivalate is replaced by an equimolar amount of benzyl bromide and 4- nitrobenzyl bromide, respectively, to product benzyl penicillanate 1,1-dioxide and 4-nitrobenzyl penicillanate 1, 1-dioxide, respectively.

EXAMPLE 10 :

Penicillanio Acid 1d-Qxide

To 1.4 g. of prehydrogenated 5X palladium- on-calcium carbonate in 50 ml. of water was added a solution of 1.39 g.0of benzyl 6,6- dibromopenicillanate Ik-oxide in 50 ml. of tetrahydydrofuran. The mixture was shaken under an . 15 atmosphere of bydrogen at ca. 45 p.s. 1. and 26°C. for 1 hour, and then it was filtered. The filtrate was evaporated in vacuo to remove the bulk of the tetrahydrofuran, and then the agueous phase was extracted with ether. The ether extracts were evaporated in vacug to give 0.5 8. of material which appeared to be largely benzyl penicillanate 1o- . oxide.

The above benzyl penicillanate lo-oxide was combined with a further 2.9 g. of benzyl 8,8 dibromopenicillanate 1& oxide and dissolved in 50 ml. of tetrahydrofuran. The solution was added to

CT 2 4.9 g. of 5% palladium-on-calcium carbonate, in 50 ml. of water, and the resulting mixture was shaken under at atmosphere of hydrogen, at ca. 45 p.a.1. and 25°C. overnight. The mixture was filtered, and b the filtrate was extracted with ether. The extracts wore evaporated in vacuo, and the residue was purified by chromatography on allica gel, eluting with chloroform. This afforded 8.50 g. of material.

The latter material was hydrogenated at ca. 45 p.s.i. at 25°C. In water-methanol (1:1) with 0.59 g. of 5% palladium-on-calcium carbonate for 2 hours.

At this point, an additional 0.50 g. of BX . ’ palladium -on-calcium carbonate was added and the hydrogenation was continued at 45 p.s.i. and 25°C.

Catt ee "8 overnight. Tha réactlbn mixture was filtered, " vo extracted with ether and the extracts were © discarded. Th: residual aqueous phase was adjusted to pil 1.5 and then extracted with ethyl acetate.

The ethyl acetate extracts were dried (Nagy504) and then evaporated in vacua to glve 9.14 g. of penicillanic acid ld-oxide. The NMR spectrum (CDC143/DM50-dg) showed absorptions at 1.4 (s, au), 1.64 (s, 3H), 3.60 (m, 2H), 4.3 (=, 1H) and 4.54 (m, ) iH)ppm. The IR spectrum of the product {Kbr disc) shown absorption at 1795 and 1745 om V. 53

BAD ORIGINAL 2 vv ol?

EXAMPLE 11

Penicillanic Acid la-Oxide

To 1.86 g. of prehydrogenated 6X palladium-- on-calcium carbonate in 30 ml. of water is added a : solution of 1.8 g. of 68,8-dibromopenicillanic acid : ld-oxide. The mixture is shaken under an atmosphere of hydrogen, at ca. 45 p.s.i. and 26°C., for 1 hour.

The reaction mixture is then filtered and the filtrate is concentrated in vacuo to remove the methanol. The residual aqueous phase is diluted with an equal volume of water, adjusted to pH 7, and : washed with ether. The aqueous phase is then avldified to pll 2 with dilute hydrochloric acid and extracted with ethyl acetate. The ethyl acetate " 1b extracts are dried (Na,y504) and evaporated in vacuo to give penicillanic acid ld-oxide.

EXAMELE 12

Penicillanic Acid 1p-Oxide

To a stirred solution of 2.65 g. (12.7 mmole) of penicillanic acid in chloroform at °C. was added 2.58 g. of B5% pure 3-chloroperbenzoic - "acid. After 1 hour, the reaction mixture was filtered and the filtrate was evaporated in vacuo.

The residue was dissolved in a small amount of chloroform. The solution was concentrated slowly 54 \ I

— (Ww a until a precipitate began to appear. At this point the evaporation was stopped and the mixture was diluted with ether. The precipitate was removed by filtration, washed with ether and dried, to give 0.815 g. of penicillanic acid lb-oxide, m.p. 140- 3°C. The IR spectrum of the product (CHCl 4 } solution) showed absorptions at 1775 and 1720 em 1.

The NMR spectrum (CDC143/MM50-dg) showed absorptions at 1.35 (a, 3H), 1.76 (s, 3H), 3.36 (m, 2H), 4.50 (s, 1H) and 5.05 (m, 1H)ppm. From the NMR spectrum, the product appeared to be ca. 90X pure.

Examination of the chloroform-ether mother liquor revealed that it contained further penicillanic acid 1p-oxide, and also some teen ome wo 18.0 penicillanic acid loroxide. Cee

EXAMPLE 13

Esterification of penicillanic acid lo- oxide or penicillanic acid 1p-oxide, as appropriate, with the requisite alkanoyloxy chlorides, according to Example 5, provides the following compounds: acetoxymethyl penicillanate la-oxide, propionyloxymethyl penicillanate la-oxide, pivaloyloxymethyl penicillanate 1d-oxide, acoetoxymethyl penicillanate 1p-oxide,

oo Vv of propionyloxymethyl penicillanate tp-oxide and pivaloyloxymethyl penicillanate 1p-oxide, respectively.

EXAMPLE 14

Reaction of penijcillanic acid lot-oxide or penicillanic acid 1p-oxide with 3-bromophthalide, 4- bromocrotonolactone or 4A-bromo--Y-butyrolactone, as appropriate, affords the following compounds: 3-phthalidyl penicillanate lot-oxide 4-crotonolactonyl penicillanate ld-oxide, 3-phthalidyl penicillanate 1B-oxide, 4-crotonolactonyl penicillanate 1p-oxide and

J-butyrolacton-4-yl penicillanate 1p oxide, respectively.

EXAMPLE 15 ’ Reaction of penicillanic acid loi-oxide or penicillanic acid 1g oxide, as appropriate, with the - requisite il-chloroalkyl alkyl carbonate or 1- (alkanoyloxy)ethyl chloride, according to the ’ 20 procedure of Example 7, provider the following. . i compounds: 1-(ethoxycarbonyloxy)ethyl penicillanate lg-oxide, methoxycarbony loxymethyl penicillanate lat-oxide, . ethoxycarbonyloxymothyl penilcillanate 1lot-oxide, 56 {

BAD ORIGINAL 9

— 4° isobutoxycarbonyloxymethyl penicillanate lo-oxide, 1- (me thoxycarbonyloxy Jethyl penicillanate 1ot-oxide, 1-(butoxycarbonyloxy)ethyl penicillanate 1o-oxide, 1-(acetoxy)ethyl penicillanate ig-oxide, 1-(butyryloxy)ethyl penicillanate 1ot-oxide, 1-(pivaloyloxy)ethyl penicillanate lo--oxide, 1-(ethoxycarbonyloxy)ethyl penicillanate 1p-oxide, methoxycarbonyloxymethyl penicillanate 1p-oxide, ethoxycarbonyloxymethyl penicillanate 1h-oxsde, i{msobutoxycarbonyloxymethyl penicillanate 1p-oxide, 1- (methoxycarbonyloxy)ethyl penicillanate 1p-oxide, 1-(butoxycarbonyloxy ethyl penicillanate 1p-oxide, 1-(acetoxy)ethyl penicillanate 1p-oxide, 1-(butyryloxy)ethyl penicillanate 1h-oxide and

LL 16 1-(pivaloyloxy)lethyl penicillanate 1f-oxide, respectively. EE or or

EXAMPLE 16

Reaction of penicillanic acid lo-oxide and penicillanic acid 1p-oxide with benzyl bromide; according to the procedure of Example 4, produces benzyl penicillanate 1o-oxide and benzyl penicillanage 1fp-oxide, respectively.

In like manner, reaction of penicillanic acid 1lot-oxide and penicillanic acid 1p-oxide with 4- nitrobenzyl bromide, according to the procedure of

RT ob . Example 4, produces 4-nitrobenzyl penicillanate lo oxide and 4-nitrobenzyl renicillanate 1p-oxide, respectively.

EXAMPLE 17

Penicillanic Acid 1.1-Dioxide

To 2.17 g. (10 mmole) of penicillanic acid "lokoxide in 30 ml. of ethanol-free chloroform at ca. : 80°C. is added 1.73 g. (10 mmole) of 3I-- chloroperbenzoic acid. The mixture is stirred for 1 hour at ca. 08°C. and then for an additional 24 hours at 26°C. The filtered reaction mixture is evaporated in vacuo to give penicillanic acid 1,1-

Cor Co dioxide. oo eT Tym - Co - EXAMPLE 18

The procedure of Example 17 is repeated, except that the penicillanic acid lg-oxide used therein is replaced by: penicillanic acid 16-oxide, acetoxymethyl penicillanate lo--oxide, ¢ 20 propionyloxymethyl penicillanate la-oxide, rlvaloyoxymethyl penicillanate lof-oxide, acetoxymethyl penicillanate 1f-oxide, propionyloxymethyl penicillanate 1p-oxide, pivaloyloxymethyl renicillanate 1p-oxide,

ee . 4 3-phthalidyl penicillanate 1o-oxide, 3-phthalidyl penicillanate 1h-oxide, 3- (ethoxycarbonyloxy)ethyl penicillanate jo-oxide, ethoxycarbony loxymethyl penicillanate 1a-oxide, . 5 sthoxycarbony Loxymethyl penicillanate 1a--oxide, {sobutoxycarbony loxymethyl penicillanate jo-oxide, (methoxycarbony loxy ethyl penicillanate 1x-oxide, - (butexycarbony Loy Jetbyl penicillanate 1or-oxide, 1-(acetoxy)ethyl ponicillanate lo-oxide, 1- (bmtyryloxy)ethyl penicillanate jto—oxide, {-(pivaloyloxy)ethyl penicillanate to—-oxide, 1 (ethoxycarbony loxy)ethyl penicillanate 1p-oxide, sothoxycarbonyloxymethyl penicillanate 1p-oxide. othoxycarbonyloxymethyl penicillanate 1p-oxide,

Loeb CE 1 {mobutoxycarbony loxymetbyl penicillanate 1f-oxide, - (mothoxycarbony Loxy ethyl riotilanate th-oxide, TT : 1 (butoxycarbony loxy ethyl penicillanate 1p-oxide, 1-(acetoxy)ethyl penicillanate 1fp-oxide, 1 (butyryloxy)ethyl penicillanate 1f-oxide and 1—(pivaloyloxy)ethyl penicillanate 1f-oxide: respectively. This affords: penicillanic acid 1,1-dioxide, acetoxymethyl penicillanate 1,1-dioxide, propionyloxymethyl penicillanate 1,1-dioxide, valoyoxymethyl penicillanate 1,1-dioxide, acetoxymethyl penicillanate 1,1-dioxide, propionyloxymetbyl penicillanate 1,1-dioxide,

. ot pivaloyloxymethyl penicillanate 1,1-dioxide, 3-phthalidyl penicillanate 1,1l-dioxide,

3-phthalidyl penicillanate 1,1-dioxide, 1-(ethoxycarbonyloxy)ethyl penicillanate 1,1- : 5 dioxide, methoxycarbonyloxymethyl penicillanate 1,1-dioxide, ethoxycarbonyloxymethyl penicillanate 1,1-dioxide, isobutoxycarbonyloxymethyl penicillanate 1,1- dioxide, 1-(methoxycarbonyloxy)ethyl penicillanate 1,1- dioxide, 1-(butoxycarbonyloxy)ethyl penicillanate 1,1- dioxide, } 1-(acetoxy)ethyl penicillanate 1,1--dioxide,

16 1-(butyryloxy)ethyl penicillanate 1,1-dioxidse, 1-(plivaloyloxy)ethyl penicillanate 1,1-dioxide, 1-(ethoxycarbonyloxy)ethyl penicillanate 1,1-dioxide, methoxycarbonyloxymethyl penicillanate 1,1-dioxide,

ethoxycarbonyloxymethyl penicillanate 1,1-dioxide, isobutoxycarbonyloxymethyl penicillanate 1,1-dioxide,

1~-(methoxycarbonyloxy)ethyl penicillanate 1,1-dioxide, 1- (butoxycarbonyloxy)ethyl penicillanate 1,1-dioxide,

. 1-(acetoxy)ethyl penicillanate 1,1-dioxide, ’ 1-(butyryloxy)ethyl penicillanate 1,1-dioxide, and

; 25 1-(pivaloyloxy)ethyl penicillanate 1,1-dioxide, . respectively.

a

EXAMPLE 19

Oxidation of benzyl penicillanate locoxide } and benzyl penicillanate 1$-oxide with 3- chloroperbenzoic acid, according to the procedure of

Example 17, produces, in each case, benzyl penicillanate 1,1-dioxide. . In like manner, oxidation of 4-nitrobenzyl penicillanate la-oxide and 4-nitrobenzyl penicillanate 1h-oxide with 3-chloroperbenzoic acid, according to the procedure of Example 17, produces 4-nitrobenzyl penicillanate 1,1 -dioxide.

EXAMPLE 20

Dame Tn SE oo ame are Penicillanic Acid 1.1-Dioxide sR ag Sao ul cee Toe

Hydrogenolysis of 4 -nitrobenzyl 156 penicillanate 1,)-dioxide, according to the , procedure of Example 3, affords penicillanic acid 1,1-djoxide.

EXAMPLE 21 ‘

To a stirred solution of 32.76 g. (0.14 mole) of penicillanfic acid 1,1-dioxide in 450 ml. of ethyl acetate was added a solution of 25.7 8. of sodium 2-ethylhexanoate (0.155 mole) in 200 ml. of ethyl acetate. The resulting solution was stirred

: 24° for 1 hour and then additional 10% excess of sodium 2-ethylhexanoate in a small volume of ethyl acetate was added. Product immediately began to precipitate. Stirring was continued for 30 minutes and then the precipitate was removed by filtration.

It was washed pequent;ially with ethyl acetate, with 1:1 ethyl acetate-ether and with ather. The solid was then dried over phosphorus pentoxide, at ca. 0.1 mm of Hg for 16 hours at 25°C., giving 36.8 g. of the title sodium salt, contaminated with a small amount of ethyl acetate. The ethyl acetate content ° was reduced by heating to 100°C. for 3 hours under vacuum. The IR spectrum of this final product (KBr disc) showed absorptions at 1786 and 1608 cm !. The

NMR spectrum (D500) showed absorptions at 1.48 (=,

JH), 1.62 (sa, 3H), 3.35 (d of d's, 1H, Jy = 16Hz, J, = 2Hz), 3.70 (d of d's, 1H, Jy = 16Hz, Jo = 4Hz), 4.26 (a, 1H) and 6.03 (d of d's, 1H, Jy = 4Hz, Jy = 2Hz )ppm. : 20 The title sodium salt can also be prepared using acetone in place of ethyl acetate. i EXAMPLE 22

Penicillanic Acid 1,1-Dioxide

To a mixture of 7,600 ml. of water and 288 ml. of glacial acetic acid was added, portionwise, 379.5 g. of potassium permanganate. This mixture 62 ano oman: P.

- —_— 0 rt was stirred for 15 minutes, and then it was cooled to 80°C. To it was then added, with stirring, a mixture which had been prepared from 270 g. of penicillanic acid, 2680 ml, of AN sodium hydorxide b and 2,400 ml. of water (pil 7.2), and which had then been cooled to A°C. The temperature rose to 15°C. during this latter addition. The temperature of the resulting mixture was reduced to 65°C. and the stirring was continved for 39 minutes. To the reaction mixture was then added 142.1 g. of sodium bisulfite, in portions, during 10 minutes. The mixture was stirred for 10 minutes at 16°C., and _ then 100 g. of supercel (a diatomaceous earth) was , : added. After a further § minutes of stirring, the

Ce... .. 8... .;mixture was £iltered. Tp, the filtrate was added 4.0 ... . liters of ethyl acetate, and then the pil of the aqueous phase was lowered to 1.55 using 6N hydrochloric acid. The ethyl acetate layer was removed and combined with several further ethyl acetate extracts. The combined organic layer was washed with water, dried (Mg€04) and evaporated almost to dryness in vacuo. The slurry thus obtained was stirred with 700 ml. of ether at 10°C., for 20 minutes, and then the solid wan collected by filtration. This afforded 82.6 g. (26% yield) of the title compound having a melting point of 154-155.5°C. (dec.).

: (1v wv

EXAMPLE 23

Pivalovloxymethyl Penicillanate 1l.1-Dioxide

To a solution of 1.25 g. pivaloyloxymethyl penicillanate in 49 ml. of chloroform, cooled to ca. -169C., was added 6.8 g. of 3-chloroperbenzoic acid.

The mixture was stirred at ca. 15°C. for 20 minutea and then it was allowed to warm to room temperature.

Analysis of the resulting solution by NMR indicated « that it contained both the lo- and lb-oxide.

The chloroform solution was concentrated to about 26 ml. and a further 0.8 g. of 3- chloroperbenzoic acid was added. This mixture was stirred overnight at room temperature, and then all the solvent was removed by evaporation in vacuo.

The residue was redissolved in ca 4 ml. of dichloromethane and 8.4 g. of 3-chloroperbenzoic acid was added. The mixture was stirred for 3 hours . and then the solvent was removed by evaporation in vacua. The residue was partitioned between ethyl . acetate and water at pH 6.0, and sodium bisulfite was added until a test for the present of peroxides - was negative. The pH of the aqueous phame was raised to B.® and the layers were separated. The organic layer was washed with brine, dried using 26 anhydrous sodium sulfate and evaporated in vacua.

The residue was dissolved in ether and 64 x

A wl” reprecipitated by the addition of hexane. The resulting solid was recrystallized from ether to give 0.357 g. of the title compound. . The NMR spectrum of the product (CDClg) showed absorptions at 1.23 (s, 9H), 1.50 (s=s,3H), 1.67 (s,3H), 3.28 (m,2H), 4.45 (s=,1H), 5.26 (m, 1H) and 5.78 (m,2l)ppm. :

EXAMPLE 24 i-Phthalidvl Penicillanate 1.1-Dioxide

To a solution of 713 mg. of 3-phthalidyl } penicillanate in 3 ml. of chloroform was added ©.430 g. of 3-chloroperbenzoic acid at ca. 10°C. The further 6.513 g. of 3-chloroperbenzoic acid was added. The mixture was stirred for 4 hours at room ' temperature, and then the solvent was removed by evaporation in vacuo. The residue was partitioned between ethyl acetate and water at pH 6.0, and sodium bisulfite was added to decompose any remaining peracid. The pH of the aguecus phase was . raised to 8.8. The layers were separated and the organic phase was evaporated in vacua. This afforded the title compound as a foam. The NMR spectrum (CDC1g) showed absorptions at 1.82 (m,6H), oe 25 3.3(m,2H), 4.52 (p,1H), 5.23(m, 1H) and 7.63 (m, SH) ppm.

et

EXAMPLE 25 2.2.2-Trichloroethyl Peniclill anate_l,1-Dioxide

To 100 ma. of 2,%2,2 trichloveethy? penicillanate in a emall volume of chloroform was added 58 mg. of deh loraporbenzoic arid and the ‘mixture was stirred for 30 minutes. Rxamination of the reaction product at this point revealed that it : was mostly sulfuxide (The NMR spectrum (CDClg3)

Pet showed absorptions at 1.8 (e,3H), 1.77 (=,311), 3.36(m,2H), 4.65 (8,14), 4.85 (m,2H) and 6.37 (m,1H)ppm.) A further 100 mg. of 3-ch loroperbenzoic acid was added and the mixtures wan etirced : overnight . The solvent was then removed by evaporation in acne, and the residuae was 156 partitioned between ethyl acetate and water at pH 6.0. Sufficient sodium bisulfite was added to decompose the excens peracid and then the pl was ’ ralsed to B.5. The organic phase was separated, washed with brine and dried. Rvaporation in vacuo i 20 afforded 65 mg. of the title product. The NMR . gpactrum (CDCl) showed absorptions at 1.53 (a,3H1), 1.72(s,3H), 3.A7(m,2H), A.5(s, 1), 4.6 (m,1H) and

A.8 (m,?H)ppm. : £6 aad ORIGINAL Bp “ (- ee

- . Lt

EXAMPLE 26 1s rabapay. Bente Lanate tA oxide

A solution of 4-nitrobenzvl penicillanate yn chlorofor® was cooled tO about 157C- and 1

Bb equivalent of 5 cnioroperbonzoic acid was added.

The reaction mixture was stirred for 29 minutes.

Examination of the reaction mixture at this point by nuclear magnetic resonance spectroscoPy revealed vo that it contained 4-nitrobenzyl penicillenate 1- oxide. A further 1 equivalent of a-chloroperbenzolic } acid was added and the reaction mixture was atirred for 4 hours At this point 2 further 1 aquivalent of 4_chloroperbenzolc acid was added and the reaction mixture was stirred overnight. The 16 solvent was removed by evaporation. and the residue was partitioned between ethyl acetate and water at pH 8.5. The ethyl acetate layer was separated; washed with water, dried and evaporated to glve the ' crude product - The crude product wan purified by 29 chromatography on silica gel, aluting with at 1:4 mixture of ethyl acetate/chlorofor®:

The NMR spectrum of the product (CDCl]) showed absorptions ot 1.35 (a, 3H). 1.68 (8, 3H), 4.45 (m, 28); a d2 (8, 1H), 4-58 0% 1H), 6-30 (= ' 25 oH) and 7.83 (Q. AH Yppm- 67 oo

D pe

EXAMPLE 27

Penicillanic Acid 1.1--Dioxide

To 6.54 g. of 4-nitrobenzyl penicillanate " 1,1-dioxide in 30 ml. of methanol and 16 ml. of ethyl acetate was added 9.54 g. of 10X palladivm-on - carbon. The mixture was then shaken under an atmosphere of hydrogen at a pressure of about 50 ‘peig. until hydrogen uptake ceased. The reaction mixture was filtered, and the solvent removed by - 10 'svaporation. The residue was partitioned between ethyl acetate and water at PH B.5, and the water layer was removed. Fresh ethyl acetate was added and the pH was adjusted to 1.5. The ethyl acetate layer was removed, washed with water and dried, and 16 then it was evaporated in vacuo. This afforded 9.168 g. of the title compound as a crystalline solid. oo EXAMPLE 28 : Penicillanic Acid 1.1-Dioxide

A atirred solution of B12 mg. of 4- . o nitrobenzyl penicillanate 1,1-dioxide in a mixture of 6 ml. of acetonitrile and 5 ml. of water was cooled to 08°C. and then a solution of 484 mg. of sodium dithionite in 1.4 ml. of 1.8N sodium 26 hydroxide was added portionwine over several

. AL o minutes. The reaction mixture was stirred for an additional 5 minutes and then it was diluted with ethyl acetate and water at pH 8.5. The ethyl acetate layer was removed and evaporated in vacuo giving 300 me. of starting material. Fresh ethyl acetate was added to the agueous phase and the pH was adjusted to 1.5. ‘The ethyl acetate was removed, dried and evaporated in yaguo giving 59 mg. of the title compound.

EXAMPLE 29 1-Mathyl-1-(acetoxy)athyl Penicillanata L.i-Dioxide

To 2.33 g. of penicillanic acid 1,1-dioxide

Co - in 5 ml. of N,N-dimethyl formamide was added 1.9 ml. oo Ct ethyldiisopropylamine, followed by thé drbpwise = neo 16 addition of 1.37 &- of 1-methyl-1-(acetoxy)ethyl chloride, at ca 20°C. The mixture was stirred at : ambient temperature overnight and then the mixture was diluted with ethyl acetate and with water. The layers were peparated and the ethyl acetate layer was washed with water at pi 9. The ethyl acetate solution was then dried (NajSO4) and evaporated in yacua leaving 1.65 8. of crude product as an oll.

The oil solidified on standing in the refrigerator, and it was then recyrstallized from a mixture of chloroform and ether giving material having a : melting point of 96-92°C.

0 (LV ¥ yv

The NMR spectrum of the crude product (CDC14) showed absorptions nt 1.4% (a, 3M), 1.62 (8, a), 1.86 (a, 3H), 1.93 (as, IM), 2.97 (8, IM), 3.43 (m, 20), 4.3 (a, 1H) and 4.57 (m, tH)ppm. 6 EXAMPLE 30

The procedure of RExzample 20 in repeated, except that the 1 methyl 1 (acetoxy ethyl chloride im replaced by the appropriate | methyl I (alkanoyloxy)othy!l chloride, to produce the following compounds: 1-methyl-1- (propionyloxy)ethyl penicillanate 1,1- dioxide, 1-methyl--1-(pivaloyloxy)ethyl penicillanate 1,1: . , . - oe . : . L. “oa 3 } } Ca ~ yt Cr | , dioxide and 16 1-methyl -1-(hexanoyloxy)ethy! penicillanic acid . 1,1- dioxide, reapectively.

EXAMPLE 31

Penicillanic Acid L.1 Dioxide

To a ntirred solution of 1.78 g. of peniclillanic actd in water, at pil 7.5, was added 1.46 m)l. of 40% peracetic acid, followed by an . additional 2.94 ml. of 40% peracetic acid 30 minuten : later. The reaction mixture was stirred for 3 days at room temperature and then it was diluted with ethyl acetate and water. Solid sodium bisulfite wan 70 : e

BAD ORIGINAL % ;

ee _ __ added to decompose excess peracid, and then the pH was adjusted to 1.5. The ethyl acetate layer was removed, dried (Nap50,4) and evaporated in vacuo.

The residue was a 3:2 mixture of penicillanic acid 1,1-dioxide and penicillanic acid 1-oxide.

EXAMPLE 32 pivalovlaxymethyl Penicillanate 1.1-Dioxide Co . A stirred solution of 595 mg. of : pivaloyloxymethyl penicillanate 1-oxide in b ml. of ethyl acetate was cooled to ca -15°C., and 5 mg. of manganic acetylacetonate was added. To the dark oo , brown mixture thus obtained was added, during several minutes, 0.89 ml. of 40% peracetic acid in : - neo J weve mmal]l mounts over several minutes: ~AfLor 40% Tot 16 minutes the cooling bath was removed, and the mixture was stirred at ambient temperature for 3 | . daya. The mixture was diluted with ethyl acetate : and water at pH B.5, and the ethyl acetate layer was removed, dried and evaporated in vacuo. This. afforded 178 mg. of material which was shown by NMR spectroscopy to be a mixture of pivaloyoxymethyl penicillanate 1,1-dioxide and pivaloyloxymethyl 1- ; oxide. : The above material was redissolved in ethyl acetate and reoxidized using 0.9 ml. of peracetic acid and b mg. of manganic acetylacetonate, an a described above, using a reaction time of 16 hours.

The reaction mixture was worked up as described above. This afforded 1868 mg. of pivaloyloxymethyl penicillanate 1,1--dioxide.

PREPARATION A

6,6-Dibromopenicillanic Acid la-Oxide

The title compound is prepared by oxidation of 6,8-dibromopenicillanic acid with 1 equivalent of 3-chloroperbenzoic acid in tetrahydrofuran at 6 259°C. for ca. | hour, according to the procedure of

Harrison et al.. dournal of the Chemical Society (London) Perkin I, 1772 (1976). - PREPARATION B

Benzyl 6.6-Dibromopenicillanate 10 To a solution of 54 g. (0.165 mole) of 6,6- . dibromopenicillanic acid in 350 ml. of N,N- . dimethylacetamide was added 22.9 ml. (0.165 mole) of triethylamine and the molution was stirred for 49 minutes. Benzyl bromide (19.6 ml., 0.165 mole) wasn added and the resulting mixture was stirred at room temperature for 48 hours. The precipitated triethylamine hydrobromide wasn filtered off, and the filtrate was added to 1,500 ml. of ice-water, + adjusted to pH 2. The mixture was extracted with ether, and the extracts were washed successively a with saturated sodium bicarbonate, water and brine.

The dried (MgS04) ether solution was evaporated in yacuo to give an off-white solid, which was recrystallized from jsopropanol. This afforded 70.0 g. (95% yield) of the title compound, m.P. 75-168°C. . . The IR spectrum (KBr disc) showed absorptions at 1795 and 1740 em 1. The NMR spectrum (CDClg) showed } ' absorptions at 1.53 (8s, 3H), 1.58 (s, 3H), 4.50 (=, iH), 5.13 (=, 21), 6.72 (a, 1H) and 7.37 (8, S5H)ppm.

PREPARATION E

2.2.2-Trichloroethyl Penicillanate

To 403 mg. of penicillanic acid in 16 ml.

Cea Co _ of dichloromethane was added 26 mg. of

Ty of . Cy RE mms . gn ra hepa dead a REE Co. 4i1sopropylearbodiimide followed by 0.18 ml. of a 2.2,2-trichloroethanol. The mixture was stirred i overnight and then the solvent was removed by evaporation in vacuo. The crude product was purified by columm chromatography using silica gel as the adsorbent and. chloroform as the sluant.

PREPARATION C

To a stirred aolution of 13.4 g. (0.03 mole) .benzyl 6,6-dibromopenicillanate in 200 ml. of dichloromethane was added a solution of 6.12 g- 256 (0.03 mole) of 3--chloroperbenzoic acid in 100 ml. of 73 i

EE ——————e eee ree ———— - ee ——————————————em mes TTT

Go 4 dichloromethane, at ca. 90°C. Stirring was continued for 1.5 hours at ca. @°C. and then the reaction mixture was filtered. The filtrate was washed successively with 5% sodium bicarbonate and water,

Bb and then it was dried (Nas80,4). Removal of the solvent by evaporation in vacua gave 12.5 g. of the title product as an oil. The oil was induced to solidify by trituration under ether. Flltration then afforded 16.5 g. of benzyl 6,6- dibromopenicillanate lo-oxide as a solid. The IR spectrum (CHClg) showed absorptions at 1800 and 1750 cem~l. The NMR spectrum of the product (CDClg)

Co _ showed absorptions at 1.3 (ss, 3H), 1.5 (8, 3H), 4.5 (s, 1H), 5.18 (8, 2H), 5.2 (=, 1H) and 7.3 (=,

PREPARATION D

4-Nitrobenzyvl Penicillanate

Reaction of the triethylamine salt of penicillanic acid with 4-nitrobenzyl bromide; © 20 according to the procedure of Preparation B, affords 4-nitrobenzyl penicillanate. :

PREPARATION FE

3-Phthalidyl Penicillanate

To a solution of 508 mg. of peniclllanic acid in 2 ml. of N,N-dimethylformamide was added ee

Se oS 9.476 ml. of diisopropylethylamine followed by 536 mg. of 3-phthalidyl bromide. The mixture was stirred overnight and then it was diluted with ethyl acetate and water. The pH was adjusted to 3.0 and the layers were separated. The organic layer was washed with water, and then with water at pH 8.0, and then it was dried using anhydrous sodium : sulfate. The dried ethyl acetate solution was evaporated in vacuo giving 713 mg. of the title ester as an oil. The NMR spectrum (CDClg) showed absorptions at 1.62 (m,6H), 3.3 (m,2H), 4.52 (s,1H), "§.23 (m,1H) and 7.63 (m,6H).

PREPARATION G

Pivaloyloxvmethvl Penicillanate :

EE ’ : Co : Cpr Ve Wet Gee eel es co

To 3.688 g. of 6,6-dibromopenicillanic acid in 10 ml. of N,N-dimethylformamide was added 1.8 ml. of diisopropylethylamine, followed by 1.40 ml. of chloromethyl pivalate. The mixture was stirred . overnight and then it was diluted with ethyl acetate and water. The organic layer was removed and washed : successively with water at pH 3.9 and water pH 8.06. " The ethyl acetate solution was dried (NagBO4) and _then evaporated in vacuo to give pivaloyloxymethyl * 6,8-dibromopenicillanate as an amber oil (3.1 g.) which slowly crystallized.

-— — . 80

The above ester was dissolved in 100 ml. of methanol, and then 3.1 g. of 10% palladium-on-carbon and 1.31 g. of potassium bicarbonate in 20 ml. of water were added. The mixture was shaken under b hydrogen at atmospheric pressure until hydrogen uptake ceased. The reaction mixture was filtered and the methanol was removed by evaporation in vacuo. The residue was partlitioned between water and ethyl acetate at pil 8, and then the organic layer was removed. The latter was dried (Na,504) and evaporated jn vacuo to give 1.25 g. of the title compound. The NMR apectrum (CDC13) showed absorptions at 1.23 (s,9H), 1.5 (8,34), 1.67 (8,3H), oC 3.28 (m,2H), 4.45 (s,1H), 5.26 (m, 1H) and 5.78 (m,2H) ppm.

PREPARATION H

4-Nitrobenzyvl Penicillanate

To a stirred golution of 2.14 g. of pencillanic acid and 2.01 ml. of ‘ 20 ethyldiisopropylamine in 10 ml. of N,N- dimethyl formamide was added dropwise 2.36 g. of 4- nitrobenzyl bromide, at ca. 20°C. The mixture was stirred at ambient temperature overnight, and then : it was diluted with ethyl acetate and water. The : © 25 layers were separated and the ethyl acetate layer

Lo as washed with at pH 2.5, followed by water at pH - B.5. The ethyl acetate solutiou was then dried = 4 oo | 76 ee —_—_ . gD ~~ (Nay504) and evaporated in vacuo leaving 3.36 g. of the title compound.

The NMR spectrum of the product (in CDClg) : showed absorptions at 1.45 (s, 3H), 1.68 (=, 3H), 3.32 (m, 2H), 4.60 (8s, 1H), 5.23 (m, 1H), 6.26 (s, 2H) and 7.85 (a, 4H) ppm.

Fe am tT ee gE ee - pn ps Fra ae ae :

Claims (14)

-— ———— Lo (0 . 4 We Claim:

1. A penicillanic acid 1,1-dioxide derivative : "of the formula: H 0, 0 i “, ’ / OH3 ’ \ . Ci po 7 N=, . : 0 coon ot \

i ...IA i 5 a pharmaceutically-acceptable salt thereof, wherein R 18 hydrogen or an ester-forming residue readily ; hydrolyzable in vivo selected from the group cqnsisting of alkanoyloxymethyl having from 3 to 8

CC. igi eke. aaa. © -parbon atoms, 1-(alkanoyloxy)sthyl having: from 4 to". © Co - 10 9 carbon atoms, 1-methyl-1-(alkanoyloxy)ethyl having from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms, 1- (alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to B carbon atoms, 3-phthalidyl, 4- crotonolactonyl and ~butyrolacton-4-yl.

2. ‘Penicillanic acid 1,1-dloxide.

3. Sodium penicillanate 1,1-dioxide.

4. Pivaloyloxymethyl penicillanate 1,1- dioxide.

5. 3-Phthalidyl penicillanate 1,1- ‘ dioxide.

qu 410 . wv. LTB UHUAY UAL WUSLY LUAY JOLLY i onic iianaie Ayo dioxide.

7. 1-Methyl-1-(acetoxy)ethyl penicillanate 1,1- : dioxide.

8. A process for the preparation of a penicillanic acid 1,1-dioxide derivative of the formula: 0, H %, “lo CHq . 0 N “, CoOR Ia or a pharmaceutically-acceptable salt thereof, wherein R is hydrogen or an ester-forming residue readily hydrolyzable in vivo selected from the group consisting of alkanoyloxymethyl having from 3 to 8 carbon atoms, 1-(alkanoloxy)ethyl having from 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)ethyl having " from 5 to 18 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 8 carbon atoms, 1- (alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms, 3-phthalidyl, 4-— crotonolactonyl and -butyrolacton-4-yl, which comprises reacting a compound of the formula: 8 BAD ORIGINAL Pp) {

ee , —— —em (0 A % 0, 0 H ’ CH H | CH “, 8 o 3 %, 8 oF 3 ” ] 8 (Hy 7" “, N ’, °r 0 “000R 0 0] (11) (111) ALL kt nme a 5 We (Re Pate awhpderm oa : ‘HR ct GH Co Cpe ne ph ed iT coe ‘ %, 8 Sy S ! OH, - J N “, 0 | COOR (1v) wherein R is as defined above, with an oxidizing agent selected from the group consisting of sodium permanganate, potassium permanganate, 3- chloroperbenzoic acid and peracetic acid, until oxidation to the corresponding 1,1-dioxide of the formula IA, wherein R ls as defined above, is 80 ‘a

LL — — : J . ’ F080 substantially complete, and, if desired, forming a pharmaceutically-acceptable salt by reacting a compound of formula 1A, wherein R is hydrogen, with a base.

9. A process according to claim 8, in which a IE compound of formula TI or ILI is oxidized with from

9.5 to 5b molar equivalents of sodium permanganate ovr potassium permanganate at a temperature within the range of -20° to 50°C. until the reaction ie substantially complete.

10. A process according to claim 8, in which a compound of formula II or IIT im oxidized with from 1 to 4 molar equivalents of 3-chloroperbenzoic acid hn Ce or peracetic acid at a temperature within the, range . . of -20° to 50°C. until the reaction is substantially complete. }

11. A process according to claim 8, in which a compound of formula IV is oxidized with from 1 to 1e molar equivalents of sodium permanganate or potassium permanganate at a temperature within the range of -20° to 59°C. until the oxidation to the 1,1-dlioxide is substantially complete.

12. A process according to claim 8, in which a compound of formula IV is oxidized with from 2 to 8 molar equivalents of 3-chloroperbenzoic acid

— an au or peradic acid at a temperature within the range of -20° 0 509°C. until oxidation to the 1,1-dioxide is subsintially complete.

13. A process for the preparation of sodium penicilinate 1,1-dioxide which comprises, reacting penicilanic acid 1.1-dioxide with modlum 2- ‘ athy lheanoate .

14. A pharmaceutical composition comprising a pharmasutically-acceptable carrier in admixture with apenicillanic acid 1,1-dioxide derivative of formul IA as defined in claim 1 or a Co | pharmesutically-acceptable salt thereof as an activeantibacterial agent, wherein the said active Chet ei arm __Angredent is present in an amount from 20% to 95% ; “a Ce SEA Ee oe “