CA1214161A - Bis-hydroxymethyl carbonate bridged antibacterial agents - Google Patents

Abstract

BIS-HYDROXYMETHYL CARBONATE
BRIDGED ANTIBACTERIAL AGENTS
Abstract Antibacterial agents in which a penicillin and a beta-lactamase inhibitor are linked by means of a bis-hydroxymethyl carbonate bridge are of the formula

Description

BIS-H~DROXY~ETHYL CARBO~ATE
BRIDGED AMTIBACTERIAL AG~MTS

The invention relates to new chemical compounds which are o~ value as antibacterial agent~. More particularly, it relates to novel bis esters of hydrox~methyl carbonate, HOCH20COOCH20H, in which one hydroxy group i~ esterified with the carboxy group of penicillanic acid 1,1 dioxide and the other hydroxy group i~ esterified with the carboxy group of an -10 ~ ;ami~openicillin.

Penicillanlc acid 1,1 dioxide ~sulbactam) is - known from U.S. 4,234,579 to be an effective beta-lactamase i~hibitor and antibacterial agent.
In UDS~ 4/244,9~1 and BritiSh Patent Application ~oO ~,044,255 bis e~ters of ~he formula N~ ~ CH3 ~ ~ C~3 0~ " COO

(VII) O O CH

o~~ ~ N ~ ' 3 /
" COO
.
are disclosed in which sulbactam i9 coupled to k~own a~tibacterial penicillins via methanediol~ In the above ormula Rl rep~Psants ce~tain ~cyl groups of ~i~
~ .

4~

known antibacterial penicillins e.y., 2-amino-2-phenylacetyl or 2-amino-2~ hydrocyphenyl)acetyl.
In copending Canadian application Serial NoO
410,940 filed September 8, 1982 and assigned to the same assignee, compounds of formula (VII) are disclosed wherein Rl iS

R2COO- ~ _ CHCO

and R2 is certain alkyl or alkoxy groups.
In U.S. 3~928,595 antibacterial compounds are disclosed in which two penicillin or two cephalosporin molecules are bridged via a carbonate ester. These compounds are of the formula R3COOCHOCOO(~HOCOR3 where R~ is H, CE3 or C2H5 and R3 is the residue of a penicillin or cephalosporin.
Ampicillin, 6-[D- (2 amino-2-phenylacetamido)]-penicillanic acid is disclosed in U.S. 2,985,648. Amoxicillin, 6-~D-(2~amino-2-[~-hydroxyphenyl~acetamido)~penicillanic acid is known from U.S. 3,192,198 and U.S. Reissue 28,744. p-Acyl derivatives o~ amoxicillin are disclosed in U.S. 2,985,648, U.S. 3,520,876 and U.S. 4,053~360.

~,.

.

~L2~

The present i~vention provides antibacterial compou~ds o~ ~he formula (I) which are ef~iciently absorbed rom ~he ga~rointe~inal tract o~ mammals and aftex absorp~ion are rapidly ~ran~ormed into -the companent alpha-aminobenzylpenlc~llln ~e..g ampicillin ~r amoxicillin~ and peni~illanic acid 1,l-dioxide (su~bactam). Said inventio~ compounds are o~ the ~ormula Rl ~--CHCON~ <C~I3 COOCX2o ~ (I3 ~ C~3 or a pharmaceutically acceptable acid addition salt thereof, whexein Rl is hydrogen, hydroxy, formyloxy, alkanoyloxy having rom two to seven carbon atoms, alXoxycarbonyloxy having from two to seven carbon atoms or R2C6H4COO where R2 is hydrogen, alkyl having from one to ~our caxbon atoms, alXoxy ha~ing from one to four carb~n atoms, F, Cl, Br, I or CN.
Particularly pre~erred values for Rl include hydrogen, hydroxy, acetoxy, pivaloylo~y or isabutyryl ox~r .

L4~

The invention also provides valuable intermediates of the formula (II) Rl ~ - CHCONU N _ ~ 3 ~II) O O
\ / C=o wherein Rl is as defined above for compound (I) and Q is a group readily convertible to amino, preferably azido, benzyloxycarbonyl-amlno, 4-nitrobenzyloxycarbonylamino or 1-methyl-2-methoxycarbon-ylvinylamino.
The invention further provides valuable intermediates of the formula 1 ~ CH (III) wherein X is a good leaving group, perferably Cl, Br or I.

The invention provides a process for production o~ the compound (I) or a pharmaceutically acceptable acl~ addition salt thereof, the process being characterized in tha-t:
a. compounds of the -Eormulae Rl ~ CECONE1~ ~ ~ C}13 ~IX) N ~CoOZ

and O O

oF~ooz ( x ) wherein Rl is as defined above, Q is azido, benzyloxycarbonylamino, 4-nitrobenzyloxycarbonylamino or l-methyl-2-methoxycarbonylvinyl-amino; one of zl and z2 is a carboxylate salt forming cation, M, and the other is CH20COCH2X where X is Cl, Br or I; are contacted in the presence of a polar organic solvent to provide an intermediate of the formula ~ s 3~ .6~
- 5a -Rl ~ /~ OOCH O (II) O o \ / CCO
~ ~ ~ CH3 and conversion of said intermediate to the product of formula (I) by conversion of said group Q ~o an amino group, or b. a compound of the formula R ~ -CHCONH ~ CH3 o ~/COOCH O

2 \ (VIII)

3 O O
R4 ~ CH ~ =O

N
// ~' COOCH20 wherein Ql is azido, benzyloxycarbonylamino, p~nitrobenzyloxy-carbonylamino;

~L2~
- 5b -R is H, Cl, Br o~ I and R4 is Cl, Br ox I; is contacted with hydrogen in ~he presence of a hydrogenation catalyst, and if required, a compound o~ ~ormula ~I) thus~produced by any process above is converted to a pharmacelltically acceptable acid addition salt thereof.
The invention also provides a process for production o the compound (II), the process being characterized ln that:
compounds of the formulae Rl ~ CHCoNH ~ C33 (IX) O ~COOZ 1 and r COOz 2 (X) wherein Rl is as de~ined above, Q is azido, benzyloxycarbonylamino,

4 nitrobenzvloxycarbonylamino or 1-methyl-2-methoxycarbonylvinyl-amino; one of zl and z2 is a carboxylate salt forming cation, M, and the other is CH20COCH2X where X is Cl, Br or I; are o contacted in the presence o~ a polar organic solvent.

~, ~

- 5c -The invention further provides a process for production of the compound (I~I), the process beiny charaaterized ln that:
a compound of the formula O O
\ / CH
F~CH (V
O COOM

is contacted with a compound of the formula XCH2OCOOCH2X, where M is a carboxylate salt forming cation r in a reaction inert organic solvent at a temperature of from -20 to 60C.

Compounds of the formula 1 ~ (IV) R ~ CHCONH ~ ~ CE3 ~ COOC~I20COOCH2X

wnere Rl .is as defined above, Q is as defined above for compounds of formula (II), are also valuable intermediates.
This invention relates to derivatives of penicillanic acid which is represented by the following structural formula:

1--~S~

~ ",/ 3 Q COOH

., , ~2~
- 5d -In derivatives of penicillanlc acid, broken line attachement (''') of a substituent to the hicyclic nucleus indicate~ that ~he substituent is b~low the plane of the nucleus. Such a sub~tituen-t i5 said to be in the alpha conEiyuratlon. Conversely, broad line attachment ( ~ ) of a substituenk to the bicyclic nucleus indic-ates that the substituent is above the plane of the nucleus. This latter confiyuration is referred to as the beta-configuration. As used herein a solid line attachment ( ) of a substituent to the bicyclic nucleus indicates that the substituent can be in either the alpha-configuration ox -the beta-confiyuration.
Compounds of the formula (I)-(IV) are named as diesters of carbonic acid. For example, the compound of formula (I) where R is hydroyen is desiynated as 6-(2-amino 2-phenylacetamido)pen-icillanoyloxymethyl l,l-dioxopenicillanoyloxymethyl carbonate;
the compound (II) where Rl is hydroxy and Q is azido i5 designated as 6-~2-azido 2-~-hydroxyphenyl)acetamido3penicillanoylox~me-thyl l,l-dioxopenicillanoyloxyme~hyl carbonate; and the compound o~ formula (III) where X is iodo i5 desig~ated S as iodomethyl l,l-dioxopenicillanoylox~f~ethyl carbo~ate~
Additionally, throughout this ~peci1catlon, whenever reference .i~ made to compound of formula (I), (II) or (IV), if not already 90 indicated, it i~
understood ~hat ~his reers to a compound in which the substituent Rl- ~ OElCON~ , wherein Q2 is amino or Q and Rl and ~ are as defined above, has the D-configuration.
The compounds of fQrmU 1 a (I) can be prepared by many of the methods known in the art for synthesis o ester~. ~owever, the preferred general method involves salt formation by condensation of a carboxy-late sal~ with a halomethyl e~ter~ Two preferred ~uch met~ods are outlined below.

. ~

~--C~CO~ ~

'COOM ~T 'COOCH20 (VI) (III) C--O
~CCH ~O

Rl -~-CHCO~H,~ ~}3 N 'COOCH2o O O
S C}~3 O'~ COOC~

( I I ~

Re~nove protecting group from Q

(I) E~ C~cO~C~i3 ~ K3 C:Os: C~2o~ o N 'COQM
,,5~=
~)~{C~;~o tV) . r : ~ ~I ) ~(I) In the above ormulae, Rl and Q are as defined above, M i5 a carboxylate salt forming cation, preferably Wa, K or ~(C4Hg~ ca~ions and X i9 a go~d leaving group, preexably C1, Br or I.
The group Rl in the above intermediates o~
formulae (IV) and (VI) and products (II) and (I) includes those compounds wherein RL is acyloxy and alko~ycarbonyloxy as de~ined above. The carboxylic acid precursors of such intermediates (VI) can be prepared e.g., by methods des~ribed in U.S. 4,053,360 by acylation of 6-aminopenicillanic acid with the appropriate acid of the fonmula Rl- ~ -C~COOH

where Rl and Q are as define~ ab~ve, or an activated derivative thereof, e~g. the acid chloride or mixed anhydride ~ormed with ethyl chloroformate. In the res~lti~g i~vention compounds ~I) and'(II) obtàined by the abo~e reactions, Rl ha.~ the same value as the star~in~ material of fonmula tVI).
~ Alternativelyt the starting material of formula (IV) or (VI~ can b~ one in which Rl is hydroxy and the resulting intermediate o formu}a ~II) i~ sub-se~uently acylated or`alkoxycarbonylated to fo~m the correspondi~g compound of formula (II) wherein Rl is alkyl~arbonyloxy, alkoxycarbonyloxy or R2C6-H4COQ as defi~ed above.

The acylation or alkoxycar~onylation o the intexmediate of ormula (II) w~erein R1 is hydroxy and Q is as previously defi~ed can be carried ouk e.g., by reac-~ing said compound o~ ~ormula (II) with ,the appropriate acid chloride or acid anhydride. The reaction is ordinarily carried out in the presence o~
a reacti.on-inert solvent sys~em. In a typical procedure, from 0~5 to 2.0 molar equivalents, and preferably about 1 molar equivalent, ~f the appro-priate acid chloride or acid anhydride is contactedwith the starting compound of formula ~ wherein i~ hydro~y, in a reaction-inert solvent, in the presence of a ter~iary amine, at a temperature in the ~ange from -10 to 30C~ Reaction inert solvents which can be used in this acylation are chlorinated hydrocarbons, such as chloroform and dichloromethane;
eth~rs, such as die~hyl e~h~r and tetrahydrofuran;
}ow molecular weight esters, such as ethyl acetate a~d butyl acetate; low moleculax weight aliphatic ketones r such as acetone and methyl ethyl ketone:
~ertiary amides, such as ~,N-dimethylormamide a~d methylpyrrolidone, acetonitrile; and mixtur~s thereof.
The tertiary ~mine is normally used in an amount equivalent to the starting acid chloride or acid anhydride, and typical tertiary amines which can be used axe triethylamine, tributylamine, diisopropyl-ethylamine~ pyridine and 4-dime~hylaminopyridine~

In each of the reaction sequences designated as A and B, above, to form the amino-protected products o~ formula (II), the respective carboxylate salt and, e.g., halomethyl ester are contacted in approximately equimolar amoun~s in ~he presence of a polar organic solvent at a temperature of from about 0 to 80C.
and preerably from 25 to 50C. While at leas~
equimolar amounts of reactants are ordinarily employed, an excess of the carboxylate salt, up to a ten-fold molar exces~, is preferred. A wide variety of solvents can be used for this reaction, however, it is usually advantageous to use a relatively polar organic solvent to minLmize the reaction time. Typical solve~ts which can be employed include ~ dimethyl-formamide, N,N-dimethylace~amide, ~-methylpyrrolidone, dimet~ylsulfoxide, ethyl acetate, dichlorom~thane, chloroform, acetone and hexamethylphosphoric triæmide.
~he time required for the reactio~ to reach substan-~ial completio~ varies according to a number of factors, such as ~he ~lixture of the reac~ants, the reaction temperature and solvent. However, at about 25C~ reaction time~ of from about 10 minutes to about 24 hours are commonly emploved.
The desired amino-protected compound of formula (II3 is then isolated by methods well known to those ~ skill in the art. For axample, the reaction mi~ture is taken up in a water Lmmi3cible solvent, e~. ethyl acetate, chloroform or methylene diohloxide, washed with water, brine and dried. Evaporation of solvent provide~ the int2nmediate o formula ~
which can be purified, if desired, e~g. by chromato-graphy o~ silica gel.

:: :

~2~4~

The removal of the amino-protecting group from the intermediate (II) is carried out by methods well known in the art, see, e.g. Gross e-t al. in "The Peptides, A~alysis, Synthesis, Biology", Academic Press, ~ew York, N.Y., Vol. 3, 1981, but due regard mu~t be given to the lability of the beta-lactam ring and the ester linkages. For example, when Q is 1-methyl-2-methoxycar~onylvinylamino, the protecting group ~l-methyl-2-~ethoxycarbonylvinyl) can be removed ~Lmply by treating the compound of formula tII) with one equivalent of a strong aqueous acid, e.g.
hydrochloric acid, in a re~ction inert solvent, at a temperature in the range o~ from -10 to 30C. In a typical procedure, the compound of formula ~ is treated with one equivalent of hydrochloric acid in aqueous acetone. The reaction is usual}y complete within a short ~Lme, e.g~ within one hour. Then the acetone is removed by evapora-tion in vacuo, and the me~hylace~oacetate by-product is removed by extraction with ether. Finally, the compound of ormula (I) is recovered by lyophilization as its hydrochloride salt.
Compounds o ormula (II) wherein Q is a~ido, ben~ylox~carbonylamino or 4-nitrobenzyloxycarbonyl-amino can be converted to the corresponding minocompound of for~ula (I) by subjecting the compound ~II) to conditions commonly ~mployed for catalytic hydrogenolysis. The compound o~ formula (II) is stirred or shaken ~nder an atmosphere of hydrogen, or hydrogen, optionally mixed with an inert diiuent such a~ nitrogen or argon, in ~he presence of a catalytic æmount o~ a hydrogenolysis catalyst. Convenient solvents for this hyd.roge~olysis are lower-alkanols t su~h as methanol and isopropanol; ethers, such as ~2~

tetrahydrofuran and dioxan; low molecular weight e3ters, such as ethyl acetate and butyl acetate;
chlorinated hydrocarbons, such as dichloromethane and chloroform; water; and mixtures of these soLvents.
S However, it is usual to choose conditions under which the starting material is soluble. The hydrogenolysis is usuaLly carxied out at a temperature in the range from 0 to 60C. and at a pressure in the range from 1 to 10 atmospheres, preferably about 3 4 at~ospheres~
The catalysts used in this hydrogenolysis reaction are the type of agents known in ~he art ~or this kind of transformation, and typical examples are the noble metals, such as nickel, palladium, platinum and rhodium~ The catalyst is usually used in an amount from 0.5 to 5.0, and preferably about 1.O, times the weight of the compound of formula (II). It is often convenient to suspend the catalyst on an inert suppor~; a particularly convenient cataly~t is palladium suspended on an inert support such as carbon.
The compounds of the ~ormula (I) will form acid addition salts, and these acid addition salts are con~idered ~o be within the scope and purview of this invention. Said acid addition salts are prepared by stan~ard methods for penicillin compounds, for example by combining a solution of the compound of formula tI) in a suitable solvent (e~g. watar, ethyl - acetate, acetone, methanol, ethanol or b7~tanol~ with a solution containing a stoichiometric equivalent of the appropriate acid. If the sal~ precipitates, it is recovered by filtration. Alternatively, it can be recovered by evaporation o~ the solvent, or, in the case of aqueou~ solutions, by lyophilization. Of particular value are ~he sulfate, hydrochloride, hydrobromide, nitrate, phosphate, citrate, tartxate, - pamoate, perchlorate, sulfosalicylate, benzenesulfona~e, 4-toluenesulfonate and 2-naphthalenesulfonate salts~
The compounds of the formula (I), and the salts thereof, can be purified by conventional methods or penicillin compounds, eOg. recrystallization or chromatography, but due regard must be given to the lability of the beta-lactam ring systems and the ester linkages.
An alternate process for preparation of the antibac~erial compounds of formula (I) employs an intermediate of formula ~ Ql r~ CH3 N 'COOC~20 R3 ~/ C~ C=O ~ VIII) R ~ C~ ~

where Rl is as previously defined, Ql is azido, benæyloxycarbonylamino or E~ni~robenzyloxycarbonyl-amino, R3 is H, Cl, Br or I and R4 is Cl, Br or I.
The intermediate (~III) upon catalytic hydrogenation,.
e.g. by ~he me~hod described above for hydrogenolysis of azido, or ben~yloxycarbonylamino compounds of formula (II), is simul~aneously hydrogeno~yzed at the Q , R~ and~or R4 substituents to provide the inventio~
compound o formula (I).

~Z~4~6~L

The intermediates (VIII) are obtained by methods analogous to t~ose described herein for preparation of intermediates of ormula (II), but employing a R3,R4-substituted l,l-dioxopenicillanate in place of the corresponding unsubstituted l,l-dioxopenicillanic acid, its salts or derivatives of formulae (III) or (V) .
Methods for preparation o~ the requisite R3,R4-disubstitu~ed l,l-dioxopenicillanic acids and salts thereo are taught in U.S. 4,234,579; British Patent Application 2,044,255 and Belgian Patent No. 882,028.
The intermediates o~ formula (I~) are obtained, for example, as outlined below.

OCH X
OCH2X -C~CO~H ~ ~ CH3 . o 'COOM

-L~X ~VI) ~ ` ~ r R ~ -C~CO~H ~ < C~3 O '~COOCH20COOCH2X

(I~) where Rl t M, Q and X are as defined above. The reaction is carried out by contacting the starting amino-protected benzylp~nicillin satt of formula (VI) with a~ least an equimolar amount, pre~erably a molar e~cess of up to te~-fold, of a bis-halomethyl carbonate in the presence of a reaction inert organic solvent and a temperature of from a~out -20 to 60C., prefer-ably from 0 to 30C. The ~olvents w~ich can be employed succ~ssfully in this reaction include the ~ame polar organic solvents employed above for prPp~ration of interm~diates of formula (II).
The intermediate halomethyl l,l-dioxopenicillan-oyloxymethyl carbonate~ of formula (III) are prepared as described above for the intermediates of formula (IV~, but employing a salt of penicillanic acid 1,l-dioxide of formula ~V) as starting material i~ place o~ the s~arting penicillin salt of ormula (VI).
bi~-Chloromathyl ~arbonate is prepared by photo-chemical chlorination of dimethyl carbonate by the method of Rling _ al., Compt. rend., 170, 111, 234(1920), ChemO Abstr., 14, 1304 (1920). bis-Bromo~
methyl carbonate and _ -iodomethylcarbonate are o~tained from the bis-chloromethyl compound by ~ reaction with e~g. sodium bromide or potasssium iodi~e by methods well known in the art.
The carboxylate salts of ormulae t~) and (VI) are obtained from the corresponding carboxyli~ acids.
A pra~erred method ~or providing the salts wherein is an alkali metal such as sodium or potassium employs the appropriate salt of ~-ethylhexanoic acid a~ base. Typically, the carbo~ylic acid of formula (V) or (VI~ is dissolv~d in ethyl acetate, a~ equimolar amount o~ ~dium or potassium 2-ethylhe~anoate is 3~ added with stirring and the pr~cipitated salt of formula (V) or (VI) collected by filtration.

~2~

The corresponding sal~s of formula (V) or (VI) where M is tetrabutylammonium can be obtained rom the corresponding acid, e.g. by neutralization with aqueous tetrabutylammonium hydroxide in the presence of a water immiscible organic solvent, preferably chloroform. The solvent layer is separated and the product isolated by evapora~ion of solvent. Alternately, ~he sodium or potassium salts o~ formula (~) or (VI) are reacted with an equimolar amount of aqueous tetrabutylammonium hydrogen sulfate in the presence of a water immiscible solvent, the preciptiated al~ali metal bisulfate salt removed by filtration and the product isolated by evaporation of solvent.
While in the present i~vention the antibacterial compounds of ~he formula (I), as defined above, are the preferxed compounds wherein a penicillin and a beta-lactamase inhibitor are linked as a bis ester of hydroxymethyl carbonate, in a broader sense th~
invention relates to antibacterial compounds of the general formula Rl ~ ~CHCO c~3 NH2 ~ ~ CH3 RS
0~ 'COOCHO
\ -- (IX) C=O
B-COOCHo RS

where RS is hydrogen or alkyl having from one to three carbon atoms, B is the residue of cartain beta-lacta~ase inhibitors and Rl is as defined above.

6~

Examples of beta-lactamase inhibitors, B, include CH3 , ~

(a) (~) O O
\/ EI
F~CH2Y X ~< 3 where ~ is Cl or CH3COO where Xl = CL, Br or I
(c) (d) and O_ " ~CH 2OH
~ r 0~ 1 "

(e) The preferred compounds of formula (I), those above wherein the beta-lactamase inhibitor residue, B, is l,l-dio~openicillanoyl, (a), are prepared, as de~cribed above, employing hydrogenolysis me~lods.
Analogous compounds of ormula (IX~ wherein B is.
moiety (b), above are sLmilarly obtained. However, certain of the above moieties B are no~ stable to hydrogenolysis. Thus, preparation of compounds of formula (IX) w~erein B is a moiety that is unstable to hydrogenolysis conditions such as tc), ~d) or (e), above, requires the use of a protecting group which is removable by mild hydrolysis, or example, as outlined below.
-R~ CHCO~ S CH3 / ' 6 ~ ~ ~' '==J NHR ~ ~ CH3 + BC02M
.. . . . . ' C02C~120COOCH2~

(X~ -MX ' .
~ ~ ' R~ C~CONH~ ~ C~3 H O
F ~1 ~ " 3 ~ (IX) C02CH 20~
~0 (XI) BCC)OCH20 where B, M, Rl and X are as d~ined above and R6 is an ~m~no protecting group whi~h is removable by mild hydrolysis, e.g., triph~nylmethyl or an enamino group such as --19~

CH3~¢
oR7 where R7 is alkoxy having from onë to three carbon atoms or aminoO
Alternately, of course, the compounds of formula (IX~
S which are not stable to hy~rogenolysis conditions can be prepared by the following methods.

BCOOC}~20COC~2X + R --~--CHCO~E~ =C~o33M

-MX

~ r (XI)~ (IX) where B, M, ~1, X and R6 are as de~ined above.
When contemplating therapeutic use for a salt of lQ an antibacterial compound o~ ~his invention, it is necessary to use a pharmaceutically-acceptable salt;
however, salts o~her than these can be used ~or a variety o~ purposes. Such purposes include isolating and purifying particular compounds, and interconv~rting phanmaceutically-acceptable salts and their non-~alt counterparts.

~2ï~

The compounds of formula (I) and pharmaceutically acceptable acid additi~n salts thereo~ possess ln v _ antibacterial activity in mammals, and this activity can ~e demonstrated by standard techniques S for penicillin compounds. For example, the compound of formula (I3 is administered to mice in which acute infections have been established by intraperitoneal inoculation with a standardized culture of a pathogenic bacterium. Infection severity is standardized such that the mice receive one to ten times the LDloo tLDloo: the minimum inoculation req~ired to consistent-ly Xill 100 percent o~ con~rol mic~)~ At the end of the test, the activity of the compound is assessed by counting the number of survivors which have been challenged by the bacterium and also have received the i~vention compound. The compounds of formula (I~
can be administered by both the oral (p.o.) and sub-cutaneous (s.c.~ route.
The in vivo activity of the antibacterial ~0 compounds of this invention makes them suitable for the control of bacterial in~ections in mammals, including man, by bo~h the oral and parenteral mcdes o~ administration.. The compounds are usaful in the control of infections caused by susceptib}e bacteria in human subjects.

~2 ~
-21~

A compound of formula (I~ wherein Rl is other than hydrogen brea~s down to 6-(2-amino-2-~4-hydroxy-phenyl]acetamido)penicillanic acid (amoxicillin~ and penicillanic acid l,l-dioxide (sulbactam) after administration to a mammalian subject by both the oral and parenteral route. Sulbac~am then functions as a beta-lac~amase inhibitor, and it increases the anti~acterial effectiveness of the amoxicillin.
Similarly, a compound or formula (I) wharein Rl is hydrogen breaXs down to 6-(2-amino-2-phenylacetamido)-penicillanic acid (ampi~illin) and sulbactam. Thu5, the compounds of the formula (I) will find use in ~he control o~ bacteria which are s~?sceptible to a 1:1 mixture of amoxicillin and sulbactam or ampicillin and sulbactam, e.g. susceptible strains of Escherichia coli and Staphylococcus aureus.
. =
In determining whether a particular strain of Escherichia coli or Staphylococcus aureus is sen , sitive to a particular therapeutic compound of the invention, the in vivo ~est described earlier can be used, Alternatively, e~g., ~he minimum inhibitory concentration (MIC) o a 1:1 mi~ture of amo~lcillin and sulbactam or ampicillin~sulbactam can be measured.
l~e MIC's can be measured by the procedure recommended by the Interna~ional Collaborative Study on Antibiotic Sensitivity Testing ~Eri~cson and Sherris, Acta.
et Microbiolo~ia Scandinavica, Supp. 217, _ . _ Section B: 64-68 ~1~71]), which employs brain heart infu~ion (B~I) agar and the inocula replicating device. Overnight growth tubes are diluted 100 rold for use as the standard inoculum (20,000-10,000 cells in appro~imately Q.002 ml. are placed on the agar surfaca; 20 ml. of B~I agar/dish~. Twelve 2 fold ~2~

dilutions o~ the test compound are employed, with i~itial concentration of the t~st drug being 200 mcg./ml Single colonies are disregarded when xeading plates after 18 hrs. at 37C. The suscept-ibility (MIC) of the test organism is accepted as thelowest concentration of compound capable of producing complete inhibitlon of growth as judged by the naked eye.
When using an antibacterial compound of this invention, or a salt thereof, in a mammal, parti-cularly manr the compound can be administered alone, or it can be mixed with other antibiotic substances a~d/or pharmaceuticalLy-acceptable carriers or diluents~ Said carrier or diluent is chosen on the basis of the intended mode of administration. For example, when considering the oral mode of administra-tion, an antibacterial compound o this invention ~an be used in the form of tablets, capsules, lozenges, troches, powders, syrup3, eli~irs, aqueous solutions and suspensions, and the like, in accordance w~th standard pharmaceutical practice. The proportional ratio of active ingredient to carrier will naturally depend on the chemical nature, solubility and stability o the ac~ive ingredient, as well as the dosage con~emplated. In the case o tablets for oral use, carriers which are commonly used includa lactose, sodium citrate and salts of phosp~oric acid. Various di integrants such as s~arch, and lubricating agents, ~uch as magnesium stearate, sodium lauryl sul~ate and talc, are commonly used in tablets. For oral adminis-tration in capsule orm, useful diluents are lactose and high molecular weight polyethylene glycols, e.g.
polyethylene glycols having molecular weights of from ~.2~6~

200Q to 4000. When aqueous suspensions are requirea for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring agents can be add~d. For parenteral administration, which includes intramuscular, intraperitoneal, subcutaneous, and in~ravenous use, sterile solutions of the active ingredient are usually prepared, and the pH of the ~olutions are suitably adjusted and buffered. For intravenous use, the total concentration of solutes should be controlled to render the preparation isotonic.
A~ indicated earlier, the antibacterial com-pounds of this invention are of use in human subjects and the daily dosages to be used will not differ significantly from other~ clinically-used, penicillin antibiotics. The prescribing physician will ultimately determine the appropriate dose for a given human subject, and this can be expected to vary according to the age, weight, and response of the individual patient as well as the nature and the severity of the patient's symptoms. The compounds o this invention will normally be used o~aLly at dosages in the range rom 20 to about 100 mg. per kilogram of body weight per day, and parenterally at dosages rom about 10 to about 100 mg. per Xilogram of body weight per day, usually in divided dQses. In some instances it may be necessary to use doses outside these ranges.

~2~

The following examples and preparations are provided solely for further illustration. Nuclear magnetic resonance spectra (NMR) were measured for solutions in deuterated chloroform (CDC13) or deuterated dLmet~yl sulfoxide (DMSO-d6), and peak positions are reported in parts per million downfield from tetra-methylsilaneO The following abbreviations for peak shapes are used: bs, broad singlet; s, singlet; d, doublet; t, triplet; q, quartet, m, multiplet~

6~

-~5 EXAM~LE 1 bis-Iodomethyl Carbonate To a solution of 10.7 ml. (15.9 g., 0.1 mole~
bis(chloromethyl)carbonate in 400 ml. acetone was added 75 g. (0~5 mole) sodium iodide. The mixture was re~lu~ed for 2 hours under nitrogen, then lert over-~ight at room temperature. The mi~ture was filtered a~d the fil~rate was concentrated ln vacuo. Methylene chloride ~500 ml.) was added and the resulting mixture was filtered. The filtrate was concentrated to about 200 ml., 200 ml. of water was added and the aqueous phase was adjusted to p~ 7.5. Aqueous sodium thio-sulfate solution was added to rPmova iodine, the organi~ phase was separated and dried over sodium sulfate. The dried methyLene chloride solution was concentrated _ vacuo to an oil which darkened on standing. The oily product was treated with a mixture of 35 ml. he~ane and 6 ml. diethyl ether at 0C., the resulting crystals war~ filtered, washed with hexane and dried, to afford 10.0 g. ~29%) of yellowish crystalline product. M.P. 49-51C. lH-NM~ (CDC13~
ppm (delta): ~.94 singlet; infrared spectrum (~ujol) cm 1 1756, 1775~

~2~

Iodomethyl 6-~D-(2-azido~2-phenyl-acetamido)~penicillanoyloxymethyl carbonate To a cooled solution (0C.) of 2.43 g. (7.1 mmoles) of bis-iodomethyl carbonate in chloroform ~16 ml~), a solution of 2.19 g. (3.5 mmole) of tetra-butylammonium ~_CD_( 2-azido-2-ph~nylacetamido)penicil-lanate in 10 ml. chloroform was added dropwise. After the addition was completed the reaction mixture was allowed to warm to room temperature and allowed to stand at room temperature overnight. The solvent was removed in vacuo and the residue was chromatographed on silica gel, eluting with 8:1 by volume me~hylene chloride/ethyl acetate to yield 822 mg. (40~) of product. lH-~MR tCDC13) ppm (delta): 1.52 (s, 3~), 1.65 (s, 3~), 4.45 (s, lH), 5.04 ~s, 1~), 5.65 (m, 4~), 5 . 92 ( s , 2H), 7.34 (s, 5H); infrared spectrum - (C~C13): 1770 ~m 1, ~2~

6-~D (2-azido-2-phenylacetamido)~-penicillanoyloxymethyl 1,L-dioxopenicillanoyloxymethyl Carbonate ~II, Rl = H, Q = ~ ) _ -- . 3 . To a solution of 822 mg. (1.4 mmole) iodomethyl 6-CD-(2-azido-2-phenylacetamido)]penicillanoyloxymethyl carbonate in 30 ml. chloroform was added dropwise at room temperature a s~lution of 1.33 g. (2.8 mmole) tetrabutylammonium 1,1-dio~openicillanate in 30 ml.
chloroformO The raa~tion mixture wa~ stirred at room temperature overnight, concentrated in vacuo and chromatographed on silica gel. Elution with 9:1 by volume methylene chloride/ethyl acetate gave 0.51 g.
o~ product (52.6% yield). 1H~MR (CDC13) ppm (d~lta):
1.4 (s~ 3EI) I 1.5 (s~ 3H~ r 1.6 (Sr 3EI) ~ 1~64 (s~ 3E~) ~
3.42 (d, J = 3~z, 2~), 4~4 ~s, lH), 4.44 (s, lH), 4.6 ~t, J ~ 3~zl lH), 5.04 (s, 1~), 5.44-6.0 (m, 6~) r 7.35 (s, 5~), infrared spectrum (CHC13): 1775 cm 1.

-2~-When the procedure of Example 2 is carried out with the tetrabutylammonium salt o 6-~D-(2-benzyl-oxycarbonylamino-2-phenylacetamido)~penicillanate in place of the corresponding azidocillin salt, the product obtained is iodomethyl 6-[D-~2-benzyloxycar-bonylamino-2-phenylacetamido)~penicillanoyloxymethyl carbonate. When the latter compound is employed as starting material in the procedure o~ Example 3, the product obtained i 6-~D-(2-benzyloxycarbonylamino-2-phenylacetamido)~penicillanoyloxymethyl 1,l-dioxo-penicillanoylox~methyl carbonate.
Similarly, starting wtih the 6-CD-(2-~-nitro-benzyloxycarbonylamino-2-phenylacetamido)~penicillanate LS6 salt in ~he method of Example 2 and carrying the product through the method of Example 3, provides 6-~D-(2-~-nitroben~yloxycarbonylamino-2-phenylacetamido)~-penicillanoyloxymethyl L,l-dioxopenicillanoyloxymethyl carbonate.

EX~M2LE 4 . . ~.
6-CD-(2-amino~2-phenylacetamido)]~
penicillanoyloxymethyl 1,1-dioxo-penicillanoyloxymethyl carbonate t I r Rl = El ) A -~olution of 1.49 gO 6~-~D-(2-azido-2-phenyl-acetamido)~penicillanoylox~methyl l,l-dioxopenicillan-oyloxymethyl carbonate in 40 ml. methylene chloride and 20 ml. isopropanol was hydrogenated at 60 psi (4.2 kg./cm.2) in ~he presence of 1.5 g. 10% Pd/C for 30 minutes. A fur~her 1.5 g. of catalyst was added and the hydrogenation ~as continued or another 30 minutes. The catalyst was then filtered off and the filtrate was concentrated to provide a white solid residueO The residue was dissolved in tetrahydrofuran/
water (1:1), the resulting solu~ion was cooled to 0C.
and adjusted to p~ 2.5 with 0.1~ hydrochloric acid.
The t~trahydrofura~ was- evaporated i~ vacuo and the resulting aqueous solution was freeze dried to afford 680 mg. (45~) of proauct as a white solld. l~-NMR
(CDC13 + CD30D) ppm ~delta): 1.42 (s, 6H), l.S (s, 3~), 1.6 (g, 3H), 3.46 ~m, 2~), 4.4 (s, 1~), 4.43 (s, lH), 4.72 (m, lH3, 5.2 (s, lH), 5.48 (q, ~ = 4Hz, 2H),

5.82 (~ + q, J = 6~z, 4~), 7.44 (s, 5~), inrared 25 . spectrum (~ujol): 1775 cm 1.
When ~he ~orresponding benzyloxycarbonylami~o c~mpounds provided in Example 3A are employed ~s starting material in the abo~e procedure, the same title compound i9 obtained in like manner.

Chlorome-thyl 1,l-dioxo-penioillanoyloxymethyl carbonate To a mixture of 1.17 g. t5 mmole) penicilLanic acid l,l-dioxidet 50 ml. chloroform and 10 ml. water is added 40~ aqueous tetrabutylammonium hydroxide with vigorous ~irring until a p~ of 8.5 is obtained. The chloro~orm layer is separated and the aqueous phase extracted with fresh chloroform. The combined organic layers are dried and coi~centrated to a small volume (about 20 ml~).
To a solution of 1.5 g. (10 mmole) bis-chloro methyl carbonate in 15 ml~ chloroform at 0~. is added dropwise the above solution of tetra~utylammonium 1,1-dioxopenicillanate. Ater the addition is completed,the mixture is allowed to warm to room temperature and stir overnight. The chloroform is evaporated in acuo and the cruae product puriied by chromatography on 5 ilica gel~

2~

Iodomethyl l,l-dioxo-~enicillanoyloxymethyl carbonate To a solution of 3.37 g~ (10 mmole) chloromethyl S l,l-dioxopenicillanoxymethyl carbonate in SO ml.
acetone is added 7.5 g. (50 mmole) sodium iodide and the mixture is stirred overnight at room temperature.
The acetone is evaporated in vacuo, the residue partitioned between water and ethyl acetate. The aqueous phase is separated, the organic phase washed with water, brine, dried, (~a2So4) and concentrated in vacuo to give the iodomethyl compound which is purified, if desired, by chromatography on silica gel.
When the above procedure is repeated but using dimeth~Lformamide as solvent in pla~e o acetone and sodium bromide in place of sodium iodide, bromomethyl l,l~dioxopenicillanoylox~methyl carbonatP is obtained.

6-[D~ methyl-2-methoxycarbonyl-vinylamino]-2-~p-hydroxyphenyl]-acetamido)]penicillanoyloxymethyl 5- l,l-dloxopenicillanoyloxymethylcarbonate (II, Rl = OH, Q = ~HCH(CH3? = C~C2CH3]
A. To 300 ml. of dichloromethane is added 41.9 g. of 6-t2-amino-2-~ A -hydroxyoheny~acet~lido)penicillanic acid trihydrate and 50 ml. of water, and then the pH
is adjusted to 805 using 40% aqueous tetrabutylammonium hydroxide7 Three layers are obtained. The upper layer is removed, saturated with sodium sul~ate and then it is eætracted with dichloromethane. The extracts ~re combined with the middle layer and the lower layer, and the resul~ing mixture is evaporated in vaouo to give an oil which crystalli7ed on tritura-tio~ with aoetone. This afforded 44.6 g. of tetra-butylammonium 6 (~-amino-2-C4-hydroxyphenyl~acetamido)-penicillanate_ The a~ove salt is added to 150 ml. o methyl acetoacetate and ~he suspen~ion is heated at ca. 6~C.
un~il a clear solution is obtained ~8 minutes). The mix~ure is allowed to cool, and the solid recov~red ~y ~iltra~ion. The solid is washed with me~hyl aceto-acetate, followed by die~hyl ether, to give 49.25 g.
o~ tetrabutylammonium 6-(2~ m~thyl-2-methoxycarbonyl-~i~ylamino~-2-~4-hydroxyphenyl~a~etamido)penicillanate.
.

B. A mixture of 7.04 g. (0.01 mole) o the tetra-butylammonium salt o amoxicillin enamine obtained in Part A, 4.28 g. (0.01 mole) iodomethyl l,l-dioxo-penicillanoyloxymethyl car~onate and 65 ml. chloroform is stirred at room temperature for eight hours. The mixture is diluted with 500 ml. ethyl acetate, washed wi~h brine, ~er, brine again and dried (~a2S04).
The solvent is evaporated in vacuo, the re~idual product is dissolved in a minimal amount of ethyl a~etate and purified by chromatography on silica gel.

6-CD- (2-~mino 2-~p-hydroxyphenyl~-acetamido)3penicillanoyloxymethyl l,l-dioxopenicillanoyloxymethyl 15. . carbonate hydrochloride (I, Rl ~ ~) To 2.5 g. 6-{D-(2~ methyl-2-methoxycarbonyl-vinylamino~-2-Cp-hydroxyphenyL~acetamido)]penicillan-oyloxymethyL l,l-dioxopenicillanoyloxymethyl carbonate dissolved in 60 ml. acetone is added 33 ml. 0.1~
hydrochloric acid and the mixture is stlrred or 20 minutes at xo~m temperature. ~he aceto~e is evaporated in vacuo, the aqueous residue ex~racted ._ with ethyl ether, then with ethyl acetate. The aqueous layer is then free2e dried to aford the title hydrochloride salt~

EX~LE 9 6-~D~(2-Amino-2-phenylacetamido)~-penicillanoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate hydrochloride (I, R1 = H~
~ , , .
To 300 ml~ chloroorm is added 40.3 g. ampicillin trihydrate and 50 ml. water~ The p~ is adjusted to 8~5 with 40~ aqueous tetrabutylammonium hydroxide, and the tetrabutylammonium salt isolated and reacted with methyl acetoacetate by the method of Example 7, Part A
but using chloroform as solvent in place of dichloro-methaneO The resulting tetrabutylammonium 6`CD-t~-cl-me~hyl-2-methoxycarbonylvinylamino~-2-phenylacetamido)]-}5 penicillanats (obtained in ~2~ yield) is reacted with iodomethyl l,l-dioxopenicillanoyloxymethyl carbon~te by the method o Example 7, Part B to provide 6-~D-~2-~l-methyl-2-mPthoxycarbonylvinylamino~-2-phenylacet-amido)~penicillanoyloxymethyl 1,1-dioxopenicillanoyl-oxymethyl carbonate. Removal of the enamine protectinggroup with aqueous hydrochloric acid in acetone by the method of Example 8 provides the title compound.
The compounds of the formula below are ob-tained in liXe manner~

CECO~H ~;~ CH3 ~H2 ,~L--!~ c~3 ' COC)CH20 0~0 C=O

o~T ~ COOCEI20/
where Rl is:
CF~3C
C2~5C~
( CE~3 ) 2~HCOQ
CH3 ( CH:2 ) 3C
C~I3 ( C~ 5 COO

( C~3 ) 2CH~
( C}~3 ~ 3COC00 C~I3 ( ~2 ) 50CC)0 4-FC6H4CoO
2-ClC~H4C00 3-BrC6H4C00 4-C~C6~4Co 3 ( C~I3 ~ 2CHC6Hd,COO
4~ I3 ) 3CC6~4C
3-C~I3~6~'1`C~
4-C2~50~*~0 _ 3H70C6H~Ct:)O
3 ( ~13 ) 2CEICH20C~;H4 0 4-n-C4H90C6H4cOO
~ Q0 6-~D-(2-Amino-2~ acetoxyphenyl]-acatamido)]penicillanoyloxymethyl 1,l-dioxo-penicillanoyloxymethyl carbonate hydroc~loride (I, Rl = CH COO) _ 3 A. 6-CD-(2~ methyl-2-methoxycarbQnylvinylamino~-2-~ -aceto~yphenyl~acetamido)~penicillanoyloxymethyl l,l-dioxopenicillanoyloxymethyl carbonate_ _ 6-~D-(2~ methyl-2-mPtho2ycarbonylvinylamino]-2-~p-hydroxyphenyl~acetamido)~penicillanoyloxy~ethyl 1,l~dioxopenicillanoyloxymethyl carbonate 2.34 g.
(3 mmole), prepared by the method o ~xample 7, and 0.366 g. t3 mmole) 4-dimethylaminopyridine are dissolved in 30 ml. dichloromethane and 0.28 ml.. (3 mmole) acetic anhydride is added. .The solution is stirred for 30 minutes, diluted to 100 ml~ with dichloromethane, wa~hed with water and brine, then dried (~a~So4)..
~vaporation of solvent _ vacuo affords the title ompound..
B. To 1.9 g of the product obtained in Part A, above, dissolved in 50 ml. acetone is added 23 ml.
0..1~ hydrochloric acid. The resulting mixture is stirred for 25 minut~s at room temperature and the acetone evaporated in vacuoO The aqueous phase is wa~hed with ethyl ether, clariied by ~iltration and freeze dried to yield the title compound.

6-C~-(2-Amino-2-~p-pivaloyloxy-phenylJacetamido)~-penicillanoyLoxymethyl 1,1-dioxopenicillanoyloxymethyl carbona~e hydrochloride ~I, Rl - (CH3)3CCOO~
The title compound is obtained by repeating the procedure of Example lO, but using 0.33 g. (3 mmole) o~ pivaloyl chloride in place of the acetic anhydride in Part A. The enamine protecting group is removed from the resulting p-pivaloy}oxyphenyl ester with aqueous ~ydrochloric acid in acetone and the product i~olated as des~ribed in Part B of Example lO.
Use of isobutyryl chloride or isobutyric anhydride n the above procedure afords the corresponding compound of formula (I) where Rl is (CH3)2CHCOO.
Similarly, use of ethyl chloroformate as the acylating agent provides the corresponding compound o~
formula (I) where Rl is CH3CH~OCOO and use of fo~mic-acetic anhydride as acy}ating agent yields (I).where is ormyloxy.

,., ~ hen the procedure o~ Example 11 is repeated, but employing the appropriate acid anhydrida, acid chloride, acid bromide or chloroformate ester as acylating agen~, the following compoun~s wherein Q is N~C(CH3) =
CHC02CH3 are obtained and hydrolyzed to the corresponding compounds of the formula wherein Q is ~H2, R~ CHCONH~< CCH3 ~ COOCH20 O O C=O
\ / ' /

O ~ ~ ~COOCH~o Rl Rl C~3CH~COO CH30COO
C~3~CH2)2COO C~3CH20COO
C~3(CH2)4COO (CH3~2CHOCOO
(C~3)2C~(C~2)3COO (cH3)3cOcOO
C6H5C (C~3)3CCH~OCOO
3-FC6H4COO C~3(~2)50C
2 C~C6~4COO (C~3)2C~(C~2)30COO
4-n-C4H9C6H4C00 2-BrC6H4COO
2 C~3 6 4 4-BrC6H4CO
4 ~3C6 4 3 n C3H7C6H4 3-C2H50C~H4C00 3-n-C3H7~C6H4COO
4--(CH3)3COC6H4COO ~--IC6H~COO
4-ClC6~C00 3-FC6H4COO

.

EX~MPLE 13 6-~D-(2-~Benzyloxycarbonylamino]-2~ hydroxyphenyl~acetamido)]-penicillanoylvxymethyl l,l-dioxopenicillanoyloxy-methyl carbo~ate (II, Rl = OH, Q = ~HCbz) To 7.40 g.. (0.010 mole~ tetrabutylammonium 6-CD -(2-~benzyloxycarbonylam~no]-2-~-hydroxyphenyl]acet-amido)~penicillanate and 3.81 g. (0.010 mola) bromo-methyl l,l-dioxopenicillanoyloxymethyl carbonate is added 50 ml~ dimethylformamide and the mixture is stixr~Qd for four hours. E~hyl acetate (500 mL.) i~
added and the mix~ure washed in turn with brine, water, brin~Q again and d~ied over anhydrous sodium sulfate. ~vaporation o~ solvent in vacuo affords the crude-product whirh can be purified by chromatography on silica gel, i~ desired.
Reaction of ~ompounds o the rormula (IV) with amino-protected penicillin salts of ~he formula (~I) by ~he above procedure aords products of formula (II) in like manner, where R , Q, M and X are as derined below.

~ 2~

Rl_~>--CHCO~ <C 3 v~< CE3 ' COOM ' COOCH2 O
XC~2~=
tVI ) ( IV ) ~ ~ r R~ CHCONH~ CH3 0.0 C=O
\5~ CH3 oF~ ~ COOCH2o (II) Ri Q M X
H C~jH5CH20CONH ~a H ~--~02c6EI4c~2ocoNH K B~
EI ~3 K
(n-/ 4~g)~L~ Cl ~o N3 ( n-C4Hg ) 4~ Br EO p 2C~;EI4 ~2 C~13~00 ~ 4 2 ~ n C4~9 ) 4~ I

6~

EXAM.PLE 14 :
6-~D-~2-Amino-2-~p-hydroxyphenyl)-acetamido)]penicillanoyloxymethyl l,l-dioxopenicillanoyloxymethyl carbonate tI Rl OH) A mixture of 2.0 g. 6-~D-(2-~benzyloxycarbonyl-amino~-2-~ -hydro2yphenyl~acetamido)~penicillanoyloxy-methyl l,l-dioxopenicillanoyloxymethyl carbonate, 50 ml~ dichloromethane, SO ml. isopropanol and 2.0 g.
10% palladium-on-caxbon is hydrogenated at 3-4 atmo-sphere (3.5-4.0 kg./cm.2) until hydrogen uptake ceases. An additional 2 g. of catalyst is added and hydrogenation continued for 30 minutes~ The catalyst is xemoved by ~ ra~ion and the filtrate evaporat~d in vacuo to a~ford the product which can be purified, __ if de~ired, by chromatography on Sephadex L~ 20*.

*A registered trademark of Pharmacia Fine Chemicals, Piscataway, ~.~T~

~2~
--~2--The remaining compounds of formula (II) provided in Example 13 are converted to the corresponding alpha-amino compound of formula (I) by the method of the preceding Example.

R~ 2 ~ ~ C~3 " COOC~20 O O ` C=O
~CCEO~) o/

where Rl is as defined for the starting material of formula (II)_ PREPA~ATION A
bis-Ch1oromethyl Carbonate The method is essentially that of Kling et al., Compt. rend. 170, 111-113, 234-236 (1920); Chem.
Abst~., 14, 1304 (1920).
A soLution of 59 ml. dimethyl carbonate in 120 ml~ ~arbon tetrachloride is cooled in an ice bath.
Chlorine gas is bubbled into the solution while irradiating with a sun lamp un~il most o~ the starting material is rea~ted. The excess chlorine is displaced by nitrogen, the solvent evaporated and the residue was distilled through a short column with ractionating head at 50 mm. pressure. The desired produc~ boils at 95-100C./50 mm. Yield, 68 g, .

~L%~6~
_a4_ PREPARATION B
Tetrabutylammonium 6-(2-Benzyloxycarbonyl-am o-2-~4-hydroxyphenyl~acetamido)penicillanate To a rapidly stirred mix~ure of l.O g. of 6 (2-b~n~yloxycarbonylamino-2-~4-hydrox~phenyl]acetamido)-penicillanic acid, 30 ml~ o~ dichloromethane and 20 ml~ of water was added 40~ aqueous ~etrabutylammonium hydro~ide until a p~ of 8.0 was obtained. Stirring was continued or 30 minutes at pH 8.0 and then the layers were separated. The aqueous layer was extracted wi~h dichloromethane, and then t~ combined dichloro-methane soiutions were dried (Na2SO~) and evaporated in vacuo~ This af~orded l.l g~ o~ t~e title compound~
The ~MR spectrum (in DMSO-d6) ~howed absorptions at 0~70-lo~O (m, 34~), 2.90-3.50 (m, 8~, 3.93 (5, lH),. SolO (5~ 2H), 5.23-5.50 (m, 3~1~ 6076 (d, 2~),

7~20 (d, 2H), 7 ao ~s, 5R), 7.76 (d, lH) and 8.6 (d, lH) ppm_ -TPtrabutylammonium 6-(2-C4-nitrobenzyloxycarbonyl-amino~2-C4-hydroxyphenyl]acetamido)penicillanate is obtained from 6-(2-~4-nitrobenzyloxycarbonylamino~-2-~4-hydro~yphenyl~acetamido)penicillanic acid and tetrabutylammonium hydroxide by ~he above method.
Tatrabutylammonium 5-~D-(2-~enzyloxycarbonyl-amino-2-phe~yl)acetamido]penicillanate, tetrabutyl-ammon~um-6-~D-2 (4-nitrobenzyloxycarbonylamino)-2-phenylacetamido~penicillana~e and tetrabutylammQnium 6-~D~ a2ido-2-phenylacetamido)penicillanate ar~
~repared in like manner.

Claims (30)

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A process for production of a compound of the formula (I) or a pharmaceutically acceptable acid addition salt thereof wherein R1 is hydrogen, hydroxy, formyloxy, alkanoyloxy having from two to seven carbon atoms, alkoxycarbonyloxy having from two to seven carbon atoms or R2C6H4COO
where R2 is hydrogen, alkyl having from one to four carbon atoms, alkoxy having from one to four carbon atoms, F, Cl, Br, I or CN;
characterized in that:
a. compounds of the formulae (IX) (X) wherein R1 is as defined above, Q is azido, benzyloxy-carbonylamino, 4-nitrobenzyloxycarbonylamino or 1-methyl-2-methoxycarbonylvinylamino; one of Z1 and Z2 is a carboxylate salt forming cation, M, and the other is where X is Cl, Br or I; are contacted in the presence of a polar organic solvent to provide an intermediate of the formula (II) and conversion of said intermediate to the product of formula (I) by conversion of said group Q to an amino group, or b. a compound of the formula (VIII) wherein Q1 is azido, benzyloxycarbonylamino, p-nitrobenzyloxy-carbonylamino;
R3 is H, Cl, Br or I and R4 is Cl, Br or I; is contacted with hydrogen in the presence of a hydrogenation catalyst, and if required, a compound of formula (I) thus-produced by any process above is converted to a pharmaceutically acceptable acid addition salt thereof.

2. A process according to claim 1, wherein said contacting of Part a is carried out at a temperature of from 0 to 80°C.

3. A process according to claim 1, wherein R1 is H or OH.

4. A process according to claim 1, wherein R1 is H.

5. A process according to claim 1, wherein Part a is carried out using a compound of formula (IX) wherein R1 is OH to produce an intermediate (II) wherein R1 is OH; said intermediate (II) is acylated or alkoxycarbonylated by reaction with an acid chloride or acid anhydride to provide an intermediate of formula (II) wherein R1 is formyloxy, alkanoyloxy having from two to seven carbon atoms, alkoxycarbonyloxy having from two to seven carbon atoms or R2C6H4COO where R2 is as previously defined; and thus-acylated intermediate of formula (II) is subsequently converted to a product of formula (I).

6. A process according to claim 1, wherein Part a is carried out using a combination of a compound of formula (IX) wherein Z1 is wherein X is Cl, Br or I and a compound of formula (X) wherein Z is Na, K or N(C4H9)4 cation; or a combination of a compound of formula (IX) wherein Z1 is Na, K
or N(C4H9)4 cation and a compound of formula (X) wherein Z2 is wherein X is Cl, Br or I.

7. A process according to claim 6, wherein Q in formulae (IX) and (II) is azido, benzyloxycarbonylamino or 4-nitrobenzyl-oxycarbonylamino and Q is converted to an amino group by catalytic hydrogenolysis; or Q in formulae (IX) and (II) is 1-methyl-2-methoxycarbonylvinylamino and Q is converted to an amino group by treating a compound of formula (II) by an aqueous strong acid.

8. A process according to claim 1, 6 or 7, wherein in the formulae R1 is CH3COO, C2H5COO, (CH3)2CHCOO, CH3(CH2)3COO, CH3(CH2)5COO, CH3OCOO, (CH3)2CHOCOO, (CH3)3COCOO, CH3(CH2)5OCOO, C6H5COO, 4-FC6H4COO, 2-ClC6H4COO, 3-BrC6H4COO, 4-CNC6H4COO, 4-CH3C6H4COO, 3-(CH3)2CHC6H4COO, 4-(CH3)3CC6H4COO, 3-CH3OC6H4COO, 4-C2H5OC6H4COO, 2-n-C3H7OC6H4COO, 3-(CH3)2CHCH2OC6H4COO, 4-n-C4H9OC6H4COO or HCOO.

9. A process according to claim 6 or 7, wherein in the formulae R1 is H or OH.

10. A process according to claim 5, wherein the intermediate (II) wherein R1 is OH is acylated by an acyl group CH3CO, (CH3)3CCO or (CH3)2CHCO; or according to claim 6 or 7, wherein R1 in the formulae is CH3COO, (CH3)3CCOO or (CH3)2CHCOO.

11. A process for producing 6-[D-(2-azido-2-phenylacetamido]-penicillanoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate or 6-[D-(2-benzyloxycarbonylamino-2-phenylacetamido)]penicillanoyl-oxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate, which process comprises reacting iodomethyl 6-[D-(2-azido-2-phenylacetamido]pen-icillanoyloxymethyl carbonate or iodomethyl 6-[D-(2-benzyloxy-carbonylamino-2-phenylacetamido)]penicillanoyloxymethyl carbonate with tetrabutylammonium 1,1-dioxopenicillanate in a polar organic solvent.

12. A process for production of 6-[D-(2-amino-2-phenyl-acetamido)]penicillanoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate or a pharmaceutically acceptable acid addition salt thereof, which process comprises:
catalytically hydrogenating 6-[D-(2-azido-2-phenyl-acetamido]penicillanoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate or 6-[D-(2-benzyloxycarbonylamino-2-phenylacetamido)]-penicillanoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate produced by the process of claim 11, and if required, converting the thus obtained desired compound into a pharmaceutically acceptable acid addition salt thereof.

13. A process for producing 6-[D-(2-[1-methyl-2-methoxy-carbonylvinylamino]-2-phenylacetamido)]penicillanoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate, which process comprises reacting tetrabutylammonium 6-[D-(2-[1-methyl-2-methoxycarbonyl-vinylamino]-2-phenylacetamido)]-penicillanate with iodomethyl 1,1-dioxopenicillanoyloxymethyl carbonate in a polar organic solvent.

14. A process for production of 6-[D-(2-amino-2-phenyl-acetamido)]penicillanoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate or a pharmaceutically acceptable acid addition salt thereof, which process comprises:
contacting 6-[D-(2-[1-methyl-2-methoxycarbonyl-vinylamino]-2-phenylacetamido)]penicillanoyloxymethyl 1,1-dioxo-penicillanoyloxymethyl carbonate prepared by the process of claim 13, with aqueous hydrochloric acid, and if required, converting the thus obtained desired com-pound into a pharmaceutically acceptable acid addition salt there-of.

15. A process according to claim 12 or 14, wherein the desired product is obtained as a free base or as the hydro-chloride.

16. The compound 6-[D-(2-amino-2-phenylacetamido)]penicill-anoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate or a pharmaceutically acceptable acid addition salt thereof, whenever produced by the process of claim 12 or 14, or by an obvious chemical equivalent thereof.

17. A process for producing 6-[D-(2-[benzyloxycarbonyl-amino]-2-[p-hydroxyphenyl]acetamido)]penicillanoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate, which process comprises reacting tetrabutylammonium 6-[D-(2-[benzyloxycarbonyl-amino]-2-[p-hydroxyphenyl]acetamido)]penicillanate with bromomethyl 1,1-di-oxopenicillanoyloxymethyl carbonate in a polar organic solvent.

18. A process for producing 6-[D-(2-[1-methyl-2-methoxycar-bonylvinylamino]-2-[p-hydroxyphenyl]acetamido)]penicillanoxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate, which process comprises reacting tetrabutylammonium 6-(2-[1-methyl-2-methoxycarbonylvinyl-amino]-2-[4-hydroxyphenyl]acetamido)penicillanate with iodomethyl 1,1-dioxopenicillanoyloxymethyl carbonate in a polar organic solvent.

19. A process for production of 6-[D-(2-amino-2-[p-hydroxy-phenyl]acetamido]penicillanoyloxymethyl 1,1-dioxopenicillanoyl-oxymethyl carbonate or a pharmaceutically acceptable acid addition salt thereof, which process comprises:
catalytically hydrogenating 6-[D-(2-benzyloxycarbonyl-amino]-2-[p-hydroxyphenyl]acetamido)]penicillanoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate produced by the process of claim 17 and, if desired, converting the thus obtained desired compound into a pharmaceutically acceptable acid addition salt thereof.

20. A process for production of 6-[D-(2-amino-2-[p-hydroxy-phenyl]acetamido]penicillanoyloxymethyl 1,1-dioxopenicillanoyloxy-methyl carbonate or a pharmaceutically acceptable acid addition salt thereof, which process comprises:
contacting 6-[D-(2-[1-methyl-2-methoxycarbonylvinyl-amino]-2-[p-hydroxyphenyl]acetamido)]penicillanoxymethyl 1,1-di-oxopenicillanoyloxymethyl carbonate produced by the process of claim 18, with aqueous hydrochloric acid, and if desired, converting the thus obtained desired com-pound into a pharmaceutically acceptable acid addition salt there-of.

21. The compound 6-[D-(2-amino-2-[p-hydroxyphenyl]aceta-mido)]penicillanoyloxymethyl 1,1-dioxopenicillanoyloxymethyl carbonate or a pharmaceutically acceptable acid addition salt thereof, whenever produced by the process of claim 19 or 20, or by an obvious chemical equivalent thereof.

22. A process for production of a compound of the formula (IV) wherein X is Cl, Br or I;
characterized in that a compound of the formula (V) is contacted with a compound of the formula XCH2OCOOCH2X, where M is a carboxylate salt forming cation, in a reaction inert organic solvent at a temperature of from -20 to 60°C.

23. A process according to claim 22, wherein M is sodium, potassium or tetrabutylammonium.

24. A process for production of a compound of the formula (II) .

wherein R1 is hydrogen, hydroxy, formyloxy, alkanoyloxy having from two to seven carbon atoms, alkoxycarbonyloxy having from two to seven carbon atoms, or R2C6H4COO where R2 is hydrogen, alkyl having from one to four carbon atoms, alkoxy having from one to four carbon atoms, F, Cl, Br, I or CN; and Q is azido, benzyloxycarbonylamino, 4-nitrobenzyloxycarbonylamino or 1-methyl-2-methoxycarbonylvinylamino; characterized in that compounds of the formulae (IX) (X) wherein R1 is as defined above, Q is azido, benzyloxycarbonyl-amino, 4-nitrobenzyloxycarbonylamino or 1-methyl-2-methoxycarbony-lvinylamino; one of Z1 and Z2 is a carboxylate salt forming cation, M, and the other is where X is Cl, Br or I; are contacted in the prsence of a polar organic solvent.

25. A process according to claim 24, wherein Q is azido or 1-methyl-2-methoxycarbonylvinylamino and R1 is hydrogen or hydroxy.

26. A process for production of a compound of the formula:

(XI) wherein W is X or a group of the formula (XII) X is Cl, Br or I;
R1 is as defined in claim 1, and Q2 is NH2, azido, benzyloxycarbonylamino, 4-nitro-benzyloxycarbonylamino or 1,-methyl-2-methoxycarbonylvinylamino;
or a pharmaceutically acceptable acid addition salt thereof when Q2 is NH2;

characterized in that:
[A] when a compound of formula (XI) wherein W is X is required, a compound of the formula (V) is contacted with a compound of the formula XCH2OCOOCH2X, where M is a carboxylate salt forming cation, in a reaction inert organic solvent at a temperature of from -20 to 60°C, [B] when a compound of formula (XI) wherein W is a group of formula (XII) wherein Q2 is as defined above other than NH2 is required, compounds of the formulae (IX) and (X) (X) wherein R1 is as defined above, Q is azido, benzyloxycarbonyl-amino, 4-nitrobenzyloxycarbonylamino or 1-methyl-2-methoxycarbon-ylvinylamino; one of Z1 and Z2 is a carboxylate salt forming cation, M, and the other is where X is Cl, Br or I; are contacted in the presence of a polar organic solvent, [C] when a compound of formula (XI) wherein W is a group of formula (XII) wherein Q2 is NH2 or a pharmaceutically acceptable acid addition salt thereof is required, (a) Q2 in the product of process [B] is converted into an amino group, or (b) a compound of the formula (VIII) wherein Q1 is azido, benzyloxycarbonylamino, p-nitrobenzyloxy-carbonylamino;
R3 is H, Cl, Br or I and R4 is Cl, Br or I; is contacted with hydrogen in the presence of a hydrogenation catalyst, and if required, a product of (a) or (b) is converted into a pharmaceutically acceptable acid addition salt thereof.

27. A compound of the formula (I) as defined in claim 1 or a pharmaceutically acceptable acid addition salt thereof whenever prepared by a process of claim 1 or an obvious chemical equivalent thereof.

28. A compound of the formula (IV) as defined in claim 22 whenever produced by the process of claim 22 or by an obvious chemical equivalent thereof.

29. A compound of formula (II) as defined in claim 24 whenever produced by the process of claim 24 or by an obvious chemical equivalent thereof.

30. A compound of formula (XI) as defined in claim 26 or a pharmaceutically acceptable acid addition salt thereof, whenever produced by the process of claim 26 or by an obvious chemical equivalent thereof.