CA1175808A - 2.beta.-CHLOROMETHYL-2.alpha.-METHYLPENOM-3.alpha.-CARBOXYLIC ACID SULFONE DERIVATIVES - Google Patents

Abstract

Abstract:

2.beta.-Chloromethyl-2.alpha.-methylpenam-3a carboxylic acid sulfone and salts and esters thereof were synthesized and found to be potent inhibitors of .beta.-lactamases.

Description

~Si!~8 I. Desc~iption:

The present invention relates to 2~-chloromethyl~2~-methylpenam-3a-carboxylic acid sulfone, a pharmaceutically acceptable salt or an easily hydrolyzed ester thereof which is useul as an inhibitor o~ ~-lactamases~

The presumed association between the resistance shown by ~-lactam antibiotics to cer~ain bacteria and the ability of those bacteria to produce ~ lactama~es has led to an intensive search for ~-lact~mase i~hibitors~ Clavulanic acid is an example of such a compound presently under intensive study. Another ~ lactamase inhibitor has in it- acid ~o~m the struc~ure H ~ 3 N l 1 ~
d~
: COO~{
and is disclosed in Eu~opean Patent Application 2927 published ~uly 11, 19 79 .

The compound having the structure H H
~2N ~2Cl . COOH

is disclosed in U.S. Patents 4,036,847, 4,00g,159, 3,993,646~ 3,989,685 and 3~954~732r :L~7S~

; U.S. Patent 4,155,912 describes 2-penem-3-carboxylic acid compounds having the formula C~

and esters and salts, and see also Farmcloc Abstracts 82090A, 10336B and 44337B.

The c~mpound (under the number CP-45899) having the structure ~ ~ .
~S~H3 COOH

is an irreversibly acting ~-lactamase inhibitor with excellent solution stability. It has weak antibacterial activity and potentiate~ ~he in vitro and in vivo activities of ampicillin versus ~-lactamase-producin~ 3trains ~A. R.
Engli~h _ al., Antimi~robia} Agents and Ch~motherapy, 14, 414-419 (1978), A-~wapokee et al., J. Antibiotics 31~12), 1238-1244 (Dec. 1978) and Derwent's Farmdoc Abstracts 89627A and 73966B].

8~

It is disclose~pbyaBp~ltzer et al.,-Mutual ~ioJ
Drugs of ~-Lactam Antibiotics and ~-Lactamase Inhibitors, J. Antibiotics, 33(10), 1183-1192 ~1980) that the principle of combining ~lactam antibiotic with a ~-lactamase inhibitor in a single molecule functioning as pro-drug for the two active components is illustrated by the linked esters 3 and 4 in which ampicillin and mecillinam, respectively, are combined with the ~-lactamase inhibitor penicillanic acid sulfone. It is shown that in man these esters are excel-lently absorbed from the gastro-int2stinal tract and after absorption hydrolyzed with sLmultaneous liberation of the active components. As a result high blood and tissue levels of antibiotic and ~-lactamase inhibitor in a balanced ratio are attained. The advantages of "mutual pro-drugs" over simple combinations are discussed.

Esters 3 and 4 referred to therein have the structures ~ IH-G0-N~ ~ G-CH=N S
NH2 ~ N ~ ~
O O CO O O O "

5co--o , ~11 CO-O

., . . ... , ,.. , ., .... ... . ,.. , . ...... . ... ;, .........

~7~

. ~

It is stated in GB 2044255 published October 15, 1980 that the invention therein relates to hi~herto unknown com-pounds of the general formula 1:

H H
Rl- CH -CO - NH ~ - S
R2 ~ ~ "

C - O - CH- A

in which Rl stands for a phenyl, 4-hydroxyphenyl, 1,4 cyclohexadienyl or a 3-thienyl group; R2 represents a primary amino or a carboxy group; R3 is a hydrogen atom, or a lower alkyl, aryl or aralkyl radical, and A stands for a radical o~ a ~-lactamase inhibitor containing a ~-lactam ring as well as a car~oxy group, A being connected via the caxboxy group.

The new compounds are useful in the treatment of bacterial infections and are in particular strongly active against ~-lactamase producing bacteria. See also Farmdoc Abstracts 60773C and 60776C.

Accordingly, the present invention provides ~he acid having the formula 7~

~H ~C l or a pharmaceutically acceptable salt of said acid or an easily hydrolyzed ester of said acid.

The pharmaceutically acceptable salts referred to above include nontoxic metallic salts such as sodium, potassium, ralcium and magnesium, the ammonium salt and substituted ammonium salts, e.g. salts o~ such nontoxic amines as trialkylamines (e.g. triethylamine), procaine, dibenzylamine, N-benzyl-~-phenethylamine, l-ephenamine, N,N'-dibenzyl-ethylenediamine, dehydroabietylamine, N,N'-bis(dehydroabietyl)ethylenediamine, N-(lower)alky~l~
piperidine (e.g. N-ethylpiperidine) and other amines which have been used to form pharmaceutically acceptable salts of peniciIlins and cephalosporins. The most preferred salts are the alkali metal salts, i.e. the sodium and potassium salts, snd the ammonium salt.

As used herein the term "physiologically hydrolyzed esters" re~ers to those pharmaceutically acceptable esters known in the art to hydrolyze to the free acid ~orm m ivo.
Examples of suitable physiologically hydrolyzed esters include phenacyl, acetoxymethyl, pivaloyloxymethyl, a-acetoxyethyl, a-acetoxybenzyl, a-pivaloyloxyethyl, ~ - , .

~17SI51~

phthalidylt~-phthalidyl), indanyl(5-indanyl), methoxymethyl~
benzoyloxymethyl, a-ethylbutyryloxymethyl, propionyloxy-methyl, valerylox~methyl, ~obut~r~loxymethyl, 6-~(R)-2-amino-2-phenylacetamido]-~ 3~ -dimeth~1-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carbonyloxymethyl and 6-[(R)-2-~mino-2-p-hydroxyphenylacetEmido3-~,3-dimethyl-7-oxo-4-thia-1-azabicyclo~.2.0]heptane-2-car~onyloxymethyl.
The preferred esters are the acetoxymethyl, pivaloyloxy methyl, methoxymethyl, phthalidyl, 5-indanyl, 6-~(R)-2--amino-2-phenylacetamido]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo~3.2.0]heptane-2-carbonyloxymethyl and 6-~(R)-2-~mino-2-p-hydroxyphenylacetamido]~ dimethyl-7-oxo-4-th~a-l-a2abicyclo~.2.0~heptane-2-carbonyloxymethyl~

There is ~urther pro~ided by the present invention the process ~or the production a~ the desired product of the ac~d having the ~ormula O o . ~S~ CH2Cl N _J~
COOH

or a pharmaceutically acceptable salt of said acld which comprises the consecutive steps of a) catalytically hydrogenating, as with a precious metal catalyst such as palladium, an ester ha~ring the f ormula . - -.

~:~7S8~

. .

C~2Cl ~r" ~ CH3 C:02Rl wherein R i3 benzyl or substituted benzyl and then : b) sub~ecting the hydrogenated product to oxidation to produce said deslred acid or ~alt thereof and then, ir desired~
~ ~ c 3 es~eri~ylng said acid or salt thereof to :~ produce an easily hydrolyzed ester o~ said ~cid.

There is ~urther provided by the present ` ~ lnvention the process for the production as the desired product of the acid having the fo~mula : ~ O o ~OOH
or a pharmaceut~cally acceptable æalt of said acid which comprises the consecutive step~ of a) oxidizing, as with KMhO4, H202 or like peroxide or peracid, an ester having the formula 0~
C- OC~2CC13 O

, .
' ' . ~ ',, 5~

to produce an ester sulfoxlde havlng the formula ~ O
Br ~ CH2Cl ""'n'S~
~N--C-OC~2CC~
and then b) reacting said ester ~ul~oxide with a metal ln aci~, as w~ th zinc in glacial acetic acid, to produce æaid des~ red ~cid or æalt and then, if desired, c ) e~terifying aid acid or salt thereof l;o produce an easily hydrolyæed ester of said acid.

' ~:~L7S~

A wide variety of oxidants known ln khe art ~or the oxidation o~ sulfides to sulfones can be used.
However, particularly convenient reagents for alkali metal permanganates, e.g. potassium permanganate, and organic perac~ds, e.g. 3-chloroperbenzoic acid.

Particularly useful protecting groups for Rl are the benzyl group and substituted benzyl groups, e~pecially 4-nitrobenzyl. Benzyl and substituted benzyl groups can be removed conveniently by catalytic hydrogenation, In this case, a solution in an inert solvent of the compound of the formula A, wherein Rl is benzyl or substituted benzyl, i~ stirred or shaken under an atmosphere of hydrvgen, or hydrogen mixed with an inert diluenk such as nitro~en or argon, in ~he presence of a catalytic amount of a hydrogenation catalys~. Convenient solvents for ~his hydrogenation are lower-alkanols, such as methanol; ethers~
such as tetrahydrofuran and dioxan; low molecu~ar weight esters, such as ethyl acetate and butyl acetate; water;
: and mLxtures of these solvents. However, it is usual to choose conditions u~der which the s artinq matexial is soluble. The hydrogenation is usually carried out at a t~mperature in the range from about 0 to about 60C. and at a pre~sure in the range from about 1 to about 100 kg./cm.~.
m e catalysts used in this hydrogenation reaction are the type o~ agent~ lcnown in the art for this kir~d of trans-formation, and typical examples are the noble metalc: J such aæ nickel, palladium, platinum and rhodiumO The cataly~t i~ usually present in an amount from about 0. 01 to about 205 weight-percent, and preferably from about 0.1 to about 1.0 weight-percent, based on the compound of formula A.
It i~ often eonvenient to s~spend the catalyst on an inert support; a particularly convenient catalyst is palladium suspended on an Lnert support such as carbon. Addition-, .... ~ . . . . .. . . . .
i, . .

7~

ally it is usual to buX~er the reaction mixture in order to operate at a pH in the range from about 4 to g~ and preferably ~rom 6 to 8. Borate and phosphate buffers are commonly used. The reaction typically takes Qbout one hour.

There is further provided by the present inventlon the esters ha~ing the ~ormula . -i-. .
- -- ~ CH~
R - CR - CO -~R ~ ""C

O
S"~ C ~ Cl I ) O ~_O
O
R2 a a ~

or RI ~ wherein Rl is hydrogen or hydroxy and R~ is hydrogen, hydroxy, methyl, methox~ or chloro and ., ` '' ` ~ ' , ~s~

preferably the ester having the fo~ula CH~5 H_CO_NH~s~CH~

NH2 ~ "~

O
. ~ ~

~ , ~ ~ ", I H2 O ~0 O
uld the ester ha~ring the formula :~0~I~_CO~

( '=O

F~ 3 1 2 O C- O
g and the proce~s ~or producgng such an ester which comprises the treatment with acid of a solutlon of a compound h~ving the formula s~
! ` - 12 R - CN - CO-MH

: R5 ~Cl! ' O ~--_O

where~n R is ~ ~ ~ r~

or R6 ~ wherein R6 is hydrogen or hydroxy and R~ is hydrogen~ hydro2yS methyl, methoxy or chloro and R is alkyl~ aralkyl or ar~l, R4 is hydrogen~ alkyl, aralkyl or aryI and R5 is aIkyl, aralkyl~ aryl, aIkoxy, ara,Iko~y, aryloxy or ~ R9 where~n ~8 and R are each hydrogen, alkyl, aralkyl or aryl or~ when taken together with the nitrogen atom, are piperldino or morpholino with said treatment with acid preferably belng c~rried out ln an organic solvent such as acetone or chlorofor~ or in aqueous or partly aqueous solution and prefer~bly between pH 1 and pH 5 at room temperature.

.~

.
' ~ 13 .

.

It is further pre~erred that, in the amino-protecting group, R3 is methyl, R4 is hydrogen and R5 is methoxy, ethoxy or methyl, this requires the use of methyl acetoacetate, ethyl acetoacetate or acetylacetone.

In the removal of the a-amino-protecting group it is preferred that use be made o~ a strong mineral acid such as hydrochloric acid or o~ formic acid.

mere is further provided by the presen~ invention as a no~el in~exmediate an ester having the formula ~rF~CHzCl ~-OR

wherein R is benzyl or substituted benzyl, and preferably ~p-nitrobenzyl, and the proces~ for its production which comprises heating, preferably at reflux, a com~ound having the ~ormula Br f~
. l I
O ~C-OR

wherein R is benzyl or substituted benzyl and preferably p-nitrobenzyl~ in an ~nert, anhydrous organic solvent, preferably dioxane, in the presence of large, and preferably ~i ~ -14- -~L75;~

equimolar, amounts of a weak tertiary Qmlne, preferably quinoline~ and an acid chloride, preferably benzoyl chlor~de, un~il the reaction is substantially co~plete.

There is provided by the present invention in addition as a novel intermediate an ester having the formula .' ' O
Br~CHj! Cl fi-OR

wherein R is benzyl or ~ubstituted benzyl, and preferably p-nitrobenæyl, and the process for its production which comprises oxidizing a solut~on in an inert solvent, preferably methylene chloride~ of a compound having the formula ~ CH
O C-OR
O
wherein R i8 benzyl or ~ub~tituted be~zyl, and preferably p-nitrobenzyl, ~t about room temperature b~ the use of a peracid, pre~erably m chloroperoxybenzoic ~cid.

. ~L17~S~

. .

There is further prov~ded by the present invention as a novel intermediate the ester having the ~ormula O O
Br, N

C-O~H2~Cl~i o the process for its production ~hich comprlses oxidizing a solution ln an inert sol~ent, preferably methylene chloride, of a compound having the formula ~ N
C-OC~CCl~
O
at about room temperature by the use o~ an oxidizing ~gent such aæ KMnO4, H202 or like peroxide or, preferably a peracid, preferably m-chloroperoxybenzoic acid.
"Skell~solve B~t is a petroleum ether ~raction of b.p. 60-68C. con~i~t~ng esæentiall~ of n-hex~ne ("Skelly~olve" is a trade name of the Skelly Oil Co.)~
me following ex~mples are provided solely ~or the purpvse of illustrating the preparation o~ representa-tive compo~nds of the present inuention and are not to be construed as limiting the invention~

*Trademark :

~75~

Preearation of Potassium 2B-Chloromethy~
carboxylate Sulfone t~3L-P2013) .
O O
- 3` 1`
r,~S ~f~

O N O N ~O2--CH2~No2 o ~r ,~N3Cl _ }Sr~CN2Cl C2--C~ ~ N02 02--CH2~1o2 4 E~2 CH2Cl ~ ~/

~ 3 ~ ~~`~f, 0~ ~02H ~ C02K

.~, (BL--P2 013 ) ~l7.~

-Bromopenicillanic Acid S-Sulfoxide ~) 1. Dissolve ~0 g (37.5 mmole) ~a~bromopenicillanic acid N,N'-dibenzylethylenediamine salt [Go Cignarella et al 9 J. Org. Chcm. 27, 2668 (1962) and E. Ev~ardg Nature 201~
1124 (1964)] in 3~0 ml of methylene chloride. Agi.tate and cool to 0C.

2. Slowly add 13 ml ~156 mmole) concentra~ed hydrochloric acid into the methylene chloride solution.
me precipitation of dibenzylethylenedi~mine HCl salt (DBED~Cl) t~kes place within a minute. Stir th~ slurry at 0 5C for lV minutes.

3. Filter to remove the DBED ECl precipitate through a precoated diatomaceous earth ("Dicalite") filter.
Wash the cake with 150 ml of methylene chloride. The filtration should be completed as quickly as possible.
Avoid holding the acidic methylene chloride solution ~or a prolonged period. mere may be some filtration problems because of the fine nature of the preeipitate. Addition of filter aid to the slurry may be helpful.

4. Wash the c~mbined methylene chloride ~iltrate and wash with 60 ml of cold water. Agitate 5 minutes and discard the aqueous phase. The pH of the wash i8 2,0-2.~.

5. me methylene chloride solution containing 6Gc-bromop~icillarlic acid is co~centrated und~r reduced pressure to a ~olume of 65-80 ml. Cool and stir the solution to 5C.
*Trademark ;

- ~7~
-lB

6. With ~igorous agitation cautiously add 13 ml (86.9 mmoles) o~ 40% peracetic acid over a period of 30 minutes.. The reactlon is exothe~ic. Maintain temperature between 15-18~C with ice bath cooling. The sul~oxide begins to crystallize after 10 ml peracetic acid is added~ Cool and stlr the slurry at 0-5C ~or two hours.

7. Filter ana wash the snow whlte cake with the following se~uence: 10 ml 5C water, th~n 10 ml 0-5C
methylene chlor~de, and finally wash with 15 ml of heptane.

8. Dry the cake in 45C air o~en to constant we~ght, about 6-10 hours should be su~ficient. Extended heating may generate a trace of pinkish color. me weight of 1 is about 16.26 gm, 73.24% yield~

9. The reactio~ m~x ~nd final product may be monitored by T~C using 15 toluene/4 acetone/l acetic acid (EAC) or 8 acetone/8 methanol/~ toluene~l HAC solvent systems. The final product should be analyæed by NMR and IR.

p-Nitro~enzyl 6a-bromopenicillanatç S-Sulfoxide (2) To a solution of 12 g (0.04 mole) of 6a-bromopenicillanic ac~d s-sul~oxide ln 100 ml of acetone was ~dded 7.5 g (0.041 mole) of potassium 2~ethylhex~loate.
The salt was collected by ~iltrationg washed with cold acetone and a~r dried to yield a total o~ 10 grams. The crystalline potassium salt was dissolved in 75 ml o~
dimethylacetamide and 7.8 g (0.04 mole) o~ p-nitrobenzyl bromide was added. The solution was stirred at 23C ~or 24 hours. The mixture was diluted with 500 ml o~ water and extracted with ethyl acetate~ me ethyl-acetate layer ,. . , , , ..... ~ . .... . ~ .. . ... . .. .. .. .. .... ... .. . .

~751~

was washed four times with water and dried over ~nhydrou~
magnesium ~ulfate. m e solvent was e~aporated at 35C
(15 mm) to an oil which crystallized. ~he light tan crystals of 2 were slurried with ether and collected by filtra~ion to yield 9 g (70%) mp 124-125C dec, .
Anal- calc'd for C15H15BrN206S: C, 41.98; H, ~.05; N, 6.52 Found: C3 42.00, H~ 3.48; N, 6,98 IR(KBr): 1800(s), 1740(s), 1610(w), 1520(s), 1450(m), 1350(s), 1060(m), 740(m) cm~l. H-NMR (60 mHz 9 DMS0): ~1.22 (s~3H), 1.6 (s,3H)3 4.67 (s,lH), 5.2 (d,J~1-5 Hz~lE), 5.45 (s~2H), 5.68 (d,J~1-5 Hz,lH), 7.5-8.5 (m~4H).

p-Nitrobenz~yl 2~chloromethyl-2a;methyl-6-bromopen~m~
carboxylate (3) -A solution o~ 5 g ~0.012 mole) of p-nitrobenzyl 6a-bromopenicill~nate S-sul~oxide (2) ln 120 ml anhydrous dioxane was heated at reflux under nitrogen ~or 4 hours with 1.5 g (0.012 mole) of quinollne and 1.6 g (0.012 mole) of benzoyl chloride. m e solution was diluted with 600 ml of ~ater and extracted with ethyl acetate. The ethyl acetate extract was washed with 5% sodium bicarbonate solution, 5% phosphoric acid solution and finally with water. m e organic layer was dried over anhydrous magneslum sul~ate and evaporated to an oil at 35C (15 mm).
me oil crystalli2ed and was collected, wa~hed with ether, and ~inally with cold toluene to ~ield ~, 3.5 g (65%) mp 130-135C dec.
r C15~15Cl.BrN205S: C~ 40 . o6; Hg 3-14; N 6 23 Found: C~ 40.19; H~ 3.12; N, 6.75 IR~KBr): 1792(s), 1740(s), 1610(w), 1520(s)9 1~53(s)9 1280(m)~ 1025(w), 990(w), 750(w~ cm~l.
NMR (60 mHz, DMS0): ~ 1.45 (s,3H)~ ~.5-4.3 (m,2H), 5.05 (s~lH)~ 5.42 (s~2H), 5.5(d,J~1-5 HZ,lH)~ 5.62 (d~J~1-5 Hz,l~), 7~5-8.5 (m,4H~.

.~ .

~ -20~
a::L7~5~

.

p-Nitrobenzyl 2~-Chloromethyl-?a-methylpenam-6a-carbox~late Sulfoxide (4) . .
A solution o~ 1 g (0.0022 mole) of p-nitrobenzyl 3~-chloromethyl-2a-methyl-6a-bromopenam-~a-carboxylate ~3~ dissol~ed in 50 ml o~ methylene chloride was stirred w~th 473 mg of (0.0022 mole) of m-chloroperoxybenzoic acid.
'~he solution was stlrred ~t 2~C for 3 hours. The methylene chloride was e~aporated to 20 ml at 15 mm and ~3C and the concentrated solution wa~ d~luted w th 50 ml of heptane ("~kellysolve B1t)O m e sol~ent was decanted and the ;residue was slurried wlth ether and (4) soon crystallized yield 250 mg, 24% mp 136-137C dec.
15 14BrClN206S: C, 38.68; H, 3.02 N 6 02 ~ound: C, ~9.14; H, 3.13; N, 5.96 IR(B r): 1800(s), 1760(s), 152V(s), 1~50(s), 1200(s), 1050(m), 8~o(w), 740(w) cm 1. E-NMR (60 mHz, DMS0):
2 (s,~H), 3.8-4.5 (m,2E)3 4.97 (sglH), 5.25 (d,J~1.5 H~ 5.45 (s92H), 5.6 (d,J~1.5 Hz,lH), 7.8-8.5 (m~4H).

Potassium 2~-Chloromethyl-2a-methyl~enam-3a-carbox~late -Sulfone (5) (BL-P2013) To a solutian of 7 g (0.015 mole) of p-nitrobenzyl 2fl-chlorom~th~1-2a-methyl-6~-bromopen~m-3a-carbox~late sulfoxide (4) in 150 ml of ethyl acetate was added a suspension o~ 4 g o~ ~0% palladium on diatomaceous earth ("Celite") and 2.8 g of sodium bicarbonate in 150 ml of water. '~he mixture was hydrogenated for 3 hours at 50 psi.

~S~

The catalyst was separated by filtration and the aqueous layer was sep~rated and treated with 1.5 g o~ potassium permanganate in 50 ml o~ water. The mixture wa~ stirred f`or 1 hour and 250 mg of sodium bisulfite was added. ~he mixture was filtered and the ~iltrate was ad~us~ed to pH 2 wlth concentrated hydrochloric acid. The solution was lyophilized to give a white amorphous powder. The solid was extracted with ethyl acetate, evaporated to volume o~ 20 ml and diluted with 100 ml of heptanes ~"Skellysolve B"). Whlte, hygroscopic, sol~d 2~-chlor~methyl-2a methylpenam-3a-carboxylic acid ~ulfone was collected. m e acid was di~solved in ~cetone and treated with solid potassium 2-ethylhexano~te.
cr~stall~ne white salt precipitated to give, a~ter ltration, 170 mg o~ 5 mp >140C dec.
Anal. calc'd for C8 ~ ClKN05S~2H20: C, 28.27; H, 3.24; N, 4.12 Foun~: C, 28.27; H, 3.69; N, 3.84 IR(KBr): 17gO(s3, 1770(m), 1620(s), 1460(m), 1370(s)~
1~10(s), 1200(s), lI40(s~, 955(m), 740(m) cm 1, H-NMR
(100 mHz, D20): S1~68(s,3~ .2-~.9 tm,J~2 ~z, J~ Hz, J~6 Ez~2H)~ 4.0-4.4 (m,2H), 4.3 (s,lH), 5002 (d d~ J~ Hz)~
~2 Hz ~ lH ) .

Pi~YaloyloxyT~eth~B-Chlorcmethyl-2-methylpenam-3 ca ~oxyla~e Sulfone 2~-Chloromethyl-2~-methylpenam-3.a-carboxylic acid sulfone in dimethylformamide is treated with one eguivalent of triethylamine and stirred to e~fect solution.
Bromomethyl pivalate (1 equivalent) in dimethylform,amide , ~S~

is then added. The resulting mixture is stirred at roomtemperature. ~he mixture is then clariied by filtxation and ~he ~iltrate poured into ice w~ter. ~he separated solid is collected by filtration, washed with water and dried to give ~he ~itle ester.

me respective acetoxymethyl, methoxymethyl 9 acetonyl a~d phenacyl esters of the ~ame acid are prepared by substituting in ~he method above for the bromome~hyl pivalate used therei~ an equimolar weight of chloromethyl acetate, chloromethyl methyl ether, ehloroacetone an~ phenacyl bromide~ respectively.
. . . ~

.
_~~~
carboxvlate Sul~one BI-P2024 C~3 0 CH2Cl ~ NaI ~ H20 ~ ClCH2-O-C-C~CH3)3 aceto A
~306) eo~

(BL-P2013) ~L7 ~ 2Cl o C02CH;!-O-C~ C~3) 3 (BL-P2024 ) . .

To a stirred suspension of 14.6g (0.04B7 mol) of BI.-P2013 (5) in 200 IQl o~ ac:etone was added 4 ml of a

10% aqueolls solution o ~odium iodide and the mixture was brought to reflux on the steam ~ath. To this refluxing suspension was added 14 . 8 ml (0 .1 mol) of redistilled chloromethyl pivalate (B.P. 34C: at 7 ~ g) all at once~
The mixture was ~tirred a~ reflux for ~hree hours and then cooled to room temperature ~22C). The crys~alline solids were collected by filtration, washed with 3 x 30 ml of as:etone and the combined filtrates were evaporated in an oil in vacuo at ~:22C. The oil was then taken up in 500 ml . .
o ethyl acetate and washed once with wa~er t200 ml) and once with saturated Na2SO", while being stirred with 2 g.
of decolorizing ¢arbon with c:ooling (ice bath). A~er 20 minutes the mixtu.~e was filtered through a diatomaceous ~Dicalite) pad with suction and the pad was washed with 4 x 100 ml of ethyl acetate. ~he combined filtrates were concentrated vacuo at 22~C to an oll~ The oil was then concentrated further at about 22C and cl mm Hg to removemost of the residual chloromethyl pivalate. The remaining oil was then tritura~ed twice with 50 ml portions o~
n-pentane and ~hen le~t over the weekend in the cold room (about 10C) under n-pentane. The ~olid crystalline mass was then broken up ~o a solid powder under 40 ml of 4:1 mixture o ether-n-pentane. The product wa~ then coll~cted by filtration, washed with ether pentane (1:7 ) then pentan~ and air dried. A~ter drying in vacuo for four hours over P205 there was obtained 13.37g. of pivaloyloxyme~hyl 2B-chloromethyl-2~-me~hylpenam-3~-carboxylate sulfone (about 75% yldj M~Po 93-95C~
al. Calcd. for C14~20ClN07S: C, 4 . 3; ~, . ; , .
Found: C, 44011; H, SrO8; N, 3.85.

~ ..

me~ enam-3~-carboxylate Sulfone (BL-PZ013) To a mixture of 20 ml of n-butanol and one gram of BL-P2~13 ~5) was added wa~er, one ml at a time, with shaking sn a æep~ratorY funnel until a pale yellow solution was obtained. The elear solution was filtered through a ~lu~ed ~ilter paper and the ~lask and fil~er paper washed with abouk 10 ml o 9 r 1 n-butanol-H20 and the co~bined ~iltrates were diluted with a ~urther 20 ml of n-butanol.
The re~ulting solution wa3 placed in a round-~uttomed fla~k on the roto-~ap and evaporated under reduced pressure to approximately one half the original volume. The ~n~w white crystalline product was collected by filtration, washed , ~ , , ... ... " .. ~ . ....... . .... .. .... ..... .. ... ......

~. -25-t~7~

with 6 x 10 ml of acetone and air dried. Yield 81G mg.
After vacuum drying 6 hour~ over P205 at cl mm %g.
there was obtai~ed 800 mg M.P. 215C (dec.) (80% yld~.
Anal. Calcd. ~or CBHgClNOSSK--}~20 C, 29.67; ~, 3,39; N, 4.63;
Cl, lO.g4; KoF~H~O~ 5~ 56 ~
Found: C, 29.23; H9 3.38; N, 4.49, Cl, 10.74; ~F.H2O, 5.74.
Thi~ recrystallization proc~dure produces a cry~talline monohydrate differing from ~he tar~.~ng ~aterial which is es~n~ially anhydrous.
.... .. .

EXample 5 Il C12 ~lCH2-0-C-Cl C ~ 0-C-Cl ~
¦ ~ClS03H
~ O
ClC ~ -0-S-Cl ~; o See Ch~m~cal Abstracts 27, 24271 and 22, 3828; and GB 299064.

ClS03CH2Cl (6 ) o o ~ c) o S~ CH2Cl ~ S~ CH~Cl ~'CH ~ 'CH
o6~N
COOK ~ C~OCH2Cl .5 7 NaI
ac etone -O ~ ~0 ~S ~ CH2Cl ~ 3 O 'C,-OCH2I
O

8 ~ ~c~-co~

C~' ' C OOK
~C N
~C ~

11C _~0 ~ C ~' OCH~

9 (U.S. Patent 3,316,247) I

: .

~1~7$~

CH~
CH--CO ~ ~C~

N~ O C--O
~c 17 H
XC~ ~o I
gCH3 1~
O O

O

;
.

~cn--co~

N~2 o c=o o S~ CH2Cl I, o ~ ~c ~

, a) A ~olution o~ chloromethyl chlorosulfate (0.115 mol) in 40 ml. dichloromethane is added dropwise, keeping the reaction temperatur~ below 30C., to a ~olution of compound 5 (0.1 mol) and potassium blcarbon~te (0.3 mol) and tetrabutylammonlum hydrogen sul~ate (OoOl mol) in 200 ml. dichloromethane-water (1:1). At the end of the addition the mlxture is stirred at room temperature for ~0 minutes, the ~rganic phase is separated and the aqueous phase is extracted with dichloromethane (50 ml.).
me combined organic phage~ are drled (Na2SO~) and evaporated in varU_ to gi~e a residue which is dissolved in ether (150 ml.). Insolubl~ material is ~iltered off afte:r adding diatomaceous earth and the ~iltr~te ls evaporated in acuo to pro~ide compound 7.

b) To a suspen~ion of compound ~ (1.5 g.) in d$methyl~ormamide (12 ml.j there is ~dded 1.6 g. bis-chloromethyl sulfate and the m~xture is stirred at ro~m temperature ~or 45 minutes. A~ter dilution with ethyl acetate (50 ml.) the mixture is washed with water and then aqueous sodium bicarbonate, dried and evaporated in vacuo to leave compound 7 QS an oil.

c) To a solution of c~mpound 5 (0.005 mol) in dimethyl~ormamide (7.5 ml.) there is added tr~ethyl~mine (0.007 mol) ~nd chloroiodomethane (0.030 mol) and the mi~ture ig stirred at room temper~ture ~or four hours.
A~ter dilutlon with ethyl ~cetate (~0 ml.) the mixture i8 washed with water (3 x 10 ml.) followed by saturated aqueou~ sodium chloride (5 ml.), drled and evaporated in vacuo to leave compound 7 as an oil.

~29-d) To a mixture of compound 5 (0.15 mol) silver nitrate (0.15 mol) and silver oxide (7.5 g.) in acetonitrate ~750 ml.) there is adde~ chloroiodomethane (1.5 molj.
After stirring for 48 hour~ at room temperaturs, the silYer salts are f~ltered off and the filtrate is evaporated to dryness in vacuo. The residue is dissolved in ethyl acetate (200 ml.) and the solution is wa~hed with s~turated aqueous sodium chloride, fTltered, dried and ev~porated in vacuo to give compound 7.

Co~pound 7 and other inte~med$ates and final products of the present in~ention are purified~ if desired, by column chromatography, as on 'tSephadex" ~I20 using chloro~orm-hexane~ 65:~5 as the eluent ~or example or by silic~ gel chromatography, e.g. using Mallinckrodt CC-7 and hexane-ethyl acetate, ~:2 or e~hyl acetate-petroleum ether9 8:2 or 7:~ or 1:9 or 15:85 or ethyl acetate-n-hexane, 4:6 or 3:1 hexane-ethyl acetate~ 3:1 or 1:1 or 1:4 or cyclohex~ne-ethyl acetate, 1:1.

Thin layer chromatogr~phy ~s also useful.
"Sephadex" is cross-l~nked dextran 2-(diethylamino) ethyl 2-~[2-(diethylamino)ethyl3diethyl~mmonio~ethyl ether chloride hydrochloride epichlorhydrln (See Merck Index~ N~nth Edition, item number 7337).

A solution o~ compound ~ (0.2 mol) and sodlum iodide (0.~ mol) in ~cetone (1~0 ml. ) is stirred at room kemperature ~or 18 hours. The resulking suspension is cooled to about 0C. and ad~usted to about pH 7.2 by the addition o~ saturated ~queous sodium bicarbonate with stirring. A~ter decolorizing by titration with 0.5 M
a~ueous æodium thiosulfate, water (150 ml.) is added dropwise to the stirred mixture to precipitate solid com2ound 8 w~ich i3 collected by filtration3 washed with acetone-water 1:1 (2 x 20 ml.)~ i~oprop~nol (2 ~ 20 ml,) and ether (2 x 20 ml.) and dried.

Ampicillin is converted to compound ~ by the use o~ methyl acetoacetate in the procedures of U.S.
Patent ~,~16,247~ Then to a stirred solution of compound 2 (,57 mol) in dimethyl~ormamide (1 liter) there is added at 5C. 0.5 mol of c~mpound 8. After stirr~ng ~or 15 minutes at 5C. the reaction mixture ~ poured into an ice-cold mixture o~ ethyl acetate (4 liters) and saturated aqueous calcium chloride (2 liters) with stirrlng. m e organlc layer i3 separated, ~ashed with saturated ~queous calcium chloride (2 x 500 ml.), filtered and evaporated to about one liter in vacuo to provide a concentrated solution of compound lO. To this concentrate there is added water (500 ml.) and n-butanol (~00 ml.) and then, dropwise, 4 N hydrochloric acid with stirring until the amino-protecting group is removed to provide a solution of compound 11. After the additlon of the acid is finished ether (l liter) and water (5Q0 ml,) are ~dded to the ætirred mixture, the aqueous phase is separated and the organic phase is extracted with water (80G mlO)~ The comblned aqueous extracts are washed with ether (1 liter) and then sodium chloride (640 g.) and d~chloromethane (2 liters) are added and the mixture is stirred for 15 mlnutes. The organic phase is separated and ~he aqueous phase is extracted with dichlorometh~ne (1 1iter) and the combined organic extracts are drled(MgSO~) i~nd evaporated to about 600 ml. under reduced pres.~ure to give a concentrated 801ut~0n 0~ compound 11. Add~tion to the concentrate of 200 ml.2 but~non~ ~ollowed by cooling prec~pitate~
solld 6-~(R)-2-Amin~-2-phenylacetamido1-3,~-d~methyl-7- - -oxo-4-thia-1-azabicyclo~3.2.0Jheptane-2~carbonyloxymethyl 2~-chloromethyl-2-~-methylpenam-3a-carboxylate sulfone

(11) which ls collected by ~iltration.

, ~S~

Example 6 6-~(R)-2-Amino-2-p-hydroxyphenylacetamido~-~,3-dimethyl-7-oxo-4-thia-1-azabicyclor~2.0Jheptane-2-carbonyloxymethyl 2~-chlorGmethyl-2-a-methylpenam-~a-carboxylate sulfone haYing the formula HO~CN--CO--NN ~

NH2 1~

o .
is produced by substitut~ng amoxicillin for the ampicillin used in the procedure of Ex~mple 5.

~s~

C~2 a ~
N-Bromos~lc clnimide o c~-azo-isobutyronit:rile ~Br ~ ~O
\=/ ~O

AS t~-ught by U.5. Patent 3,860,579, recrystallized phthalide (50 g,3 0.~75 mol~ and recryætalli~ed N-bromo-succin1~ide (0.375 mol~ were re~luxed by 4.5 ho~rs ~n the presence of about 100 mgm a-azobutyronitrile ln one liter CC14. The mixture was cooled to about 15C. and ~iltered to remove succln~mide which was itself washecl with about 100 ml. CC14 and filtered. The combined CCl~
phases were concentrated in vacuo to about 150 ml. giving solid ~-bromophthalide which was collected by filtration, washed with about 50 ml. CC14 and air-dried to yield 54 g.
whlch weigh~d ~0 g. a~ter recrystallization from boili~g c~clohexane, m.p. 8~-86C.
b) O O
~ /~ CHBr ~

F~ C113~Cl , ~ o 12 C-O~

D~F
22C.

'' ~ ~ .

~1 7$8~

o o ~ 5~ CH2C

r~cH3 N ~ *
~-0-CH ~ BL-P2036 ~C ~ *dl To a stirred partial solution and partial suspension of compound 5 (BL-P2013; 2.3 g., 0.0075 mol) in 20 ml~
dim~thylformamide (DMF;. dried at least ~ weeks over 3A
molecular seives~:was added 1.7 g. ~o.oo8 mol) of ~-bromophthal~de (12) and the mixture was stirred 4 hour~
at 22Cc The resultlng mixture waæ poured lnto a mixture of 200 ml. ice-cold water and 200 ml. ice-cold ethyl acetate (rinsing the fla,sk with a 1 ttle ethyl acetate) and the mixture was shaken. m en the organic solvent phase was separated and washed with seven portions of ice-cold water (lQO.ml~). The ethyI- acetate phafie ~as ~hed o~ce w~ th saturated aqueous Na2S04, dried in the cold over Na2S04~
~lltered and evaporated to dryness in vacuo to leave as the residue an oil which was tritur~ted twlce with methylcyclo~
hexane (25 ml.), twice with "SkellyRolve ~" ~b.p. 60-68C., essentially n-hexane) t25 ml.) and ~our tlme~ with 25 ml.
n-hexane to 2.5 g. compound 13 as a nearly white solid a~ter drylng in air. Thiæ product was then dried over P205 at le5s tha~ 1 mm Hg to g~ve 2.5 g. compound ~, m.p. 104C. (dec.). Its est~mat~d purlty was 85-95~.

~L~7S~

C16H14ClN07S: C, 51.61 H 7 79 Cl, 9.53.
Found: C, 52.59; Hg 4.67, N, 3.21;
Cl, 7.73; K~FoH20~ 0.27.
.

Plvaloyloxymeth~l 2~-Chlorometh~l-?~-m~thy~enam-~a~
carboxylate Sulfone A mi~ture o~ 1 g. (0.0031 mole) of potassLum 2~-chloromethyl-2a-methylpenam~ carboxylate sulfone hydrate and 1 g. o~ ~A molecular s1eves was stirred in 15 ml. o~ dimethylacetamide for 2 hours at 23, To this mixture was added 470 mg. (0.00~1 mole) of pivaloyloxymethyl ehloride and the stirring was continued for 18 hours. The molecular sieves were collected and the filtrate was di~uted with 100 ml. of water and ~xtracted with ethyl acetate. The ethyl acetate was washed nine times with w~ter ~nd dried over anhydrous magnesium sulfate.
The solvent was removed at ~0 (15 mm) to leave an oil which was chromatographed on silica using silicar CC-7 (methylene chloride 8, eth~1 acetate 2) showing 1 ~pot Rf .5. me re~idue obtained crystallized from heptane ("Skelly~olve B") to yleld 100 mg. (M.P. 94-95) of pivaloyloxymethyl 2~-chloromethyl-2a~meth~1penam-3a-carboxylate sulfone.
Anal. Calcd.: C, 44.0~; H, 5.27; N, 3.67.
Found : C, 44.20; E, 5.24; N, ~.6~.
The NMR and IR spectra were consistent ~or structure.
. . .

~s~

Sodium 2~-Chlorometh~l-?a-me~ylpenam-~a-carboxylate Sulfone CH2Cl H+ NaEH ~ CH2Cl ~ i ~2Na C8HgClN05SNa (306) (2~g.67 _ _ _ _ _ To a stirred solution of 500 mg~ of B~P2013 (potassium salt) in 5 ml. of ~ 0 and lO ml. of eth~l acetate was added ZN HCl until pE 1 was obtained (done in an ice-bath ~ith ~igorous stirring). The mixture was : then saturated with Na2S04, the aqueous layer separated and the organic phase dried briefly ~n ice over Na~SO49 filtered and treated dropwise with 50% NaE~ (sodiu~
2-ethylhe~anoate) in ~nhydrous n-butanol until neutral to moist pH paper. Product did not crystallize upon scratching and was then concentrated ln vacuo to an oil which was dissolved in acetone (5 ~l.), scratch~d - no crystals~ ether added to cloud point - no cyrstals. C~ncentrated in vacuo on rotovap. to an oil which wa~ dissol~ed in ethyl ac~tate -added one drop ~ 0 - scratched - no cry~tals. Concentrated in ~acuo and then residue was trlturated ~ith 5 ml. of n-butanol - 200 mg. of amorphous white powder obtained, ether washed - air dried - ~acuum dr~ed over P205 for 24 hours.
180 mg. final yleld of sodium 2~-chloromethy1-2a-me~hylpenam-3a-carb~xylate sulfone; deca pt~ ~100 inde~.
Anal. Calcd. for C8H~ClN0 SNa: C, ~3.10; H, 3.13; N, 4.89 _ ~ ~ 5 Found: C~ 33.20; ~, 3.69; N, 4.44 K~FoH20~ 4~04 .

. , , .

~s~

Potassium 2~ chloromethyl-2-methylpenam-~ earboxylate Sulfone (BL-P201~) To 10 L o~ water, 130 g. (1.25 mole) o~ sodium hydrogen carbonate and 200 g. of 10 ~ Pd on BaS04 were added 272 g. (o.565 mole) o~ p-nitrobenzyl 6a-bromo-2~-chloromethyl-2-methylpenam-3~carboxylate sulfone dissolved in 5 L o~ ethyl aceta~e. The mixture was hydrogenated at 40C and 1 Kg of pressure After 5 hours, the hydrogen uptake became very slow and 200 g of 10% Pd ~n BaS04 were added and the mixture hydrogenated until no ~urther signlficant hydrogen absorption was perceptible.
.
me slurry was *iltered thr~ugh a diatomaceous earth ("Celite"~ pad, the pad was washed with water and the aqueous phase washed with ~ L of ethyl acetate. To the aqueous solution, 3 B o~ ethyl acetate were added and ~he pH o~ the mixture adjusted to 105 wlth 150 ml o~ 12 N HCl at 10C. m e org&nic phase was separated and the aqueous solution saturated with Na2S04 10 H20 and extracted with 2 x 1 L of ethyl acetate. The combined extracts were dried with magnesium sul~ate. The drying agent was removed and 2~0 ml of 2 ~ potassium 2-ethylhexanoic ~cid in butanol were added at 0C.
A~ter stirring 2 hours at 0C, the potassium 2~-chlor~methyl-2-methylpenam-3-carboxylate sul~one (BL-P201~) was collected and dried ln a vacuum at room temperature.
Yield: 1~4.8 g (about 70~).

*Trade~ark ~l~L'75 Example 11 p-Nitrobenzyl 6a-Bromo~enicillinate Sul:f oxide o (101) ~N02 N ~ C0 H (216) (296 ) /
o Br"f~"~S ~<

~M ~,~ /~\
C2CH2 ~ ~02 (431. 28) . . .
Procedure:
To 200 ml. of N,N-dlmethylacetAanide was added 44 g. (0.148 mole) of 6a-bro~openicillanic acid sulfoxlde followed by 20.5 ml. (0.148 mole) of tr~ethylamine a~d ~58.2 g. (0.17r mole) of p-nitrobenzyl bromide. It was stirred at 22 for 20 hours.
The reaction mixture was poured lnto 1 liter H20 and extracted into ~ x ~00 ml. of methylene chloride~ The combined methylene chloride extracts were washed with 200 ml.
of 5% aqueouR sodium bicarbonate solution and dried over sodium sulfate at 5 for half an hour. me solution was fil~ered and eraporated under vacuum to a residue. The -3R- ' ~S~

. .
residue was diluted with ether and the solid collécted by ~iltration to yield 54 gO p-nitrobenzyl 6a-bromopenicillinate sulfoxide a~ter drying.
~5% y~ela.
nmr consistent ~or structure.
The y~eld for this step w&S the s~me as for the K-salt esterification. me advantage is there was no need to make the E-salt. (A step which goes in 85% to 90% yield).
_ _ _ . . . .. ... . . . ............. . .
.
Ex~m~le 12 To 4.375 L of N,N-dimethylacetamide was added 873,o g (2.95 moles) of 6a-bromopenicillanlc acid (S) sulfoxide and then with stirring and while keeping the internal temperature below 35C, 293 g (2.95 mole~) of tr1ethgl~ine followed by 764 g (~.54 moles) o~ p-nitrobenzyl bromide. The mixture was stirred then at room temperature for 5 hour~ and let staQd overnight.
m e reaction mixture was poured into 20 L o~ water and extracted with ~ x 7 L Or meth~lene chloride. The comblned organic extracts were wa~hed 5 x 7 L of water and then ~ith 7 L o~ 5% &queou~ sodium bicarbo.nate solution and dried over anhydrous magnes1um sulrate.
me magnesium sulfate was ~iltered off and the solutlon evaporated to & crystalline resi~ue; 4 L o~ diethyl ether were added and the cry~tal~ collected to y~eld a~ter drying at ro~m temperature 1171 g (92~) o~ p-nitrobenz~l 6~-bromopenicillanate sul~oxide.
Br 18.48% (calculated 18.53~), $ (0.25% MeOH) + 162.
.. . ..

Preparation of P-Nitrobenz~l 6a-Bromo-2~-chlorometh~
2-methylpenam-3-carboxylate Sulfone To 16 L of a~etic acid was added ~64.6 g (0~812 mole) of p-nitrobenzyl 6a-bromo 2~-chloromethyl-2-methylpenam-~-c~rboxylate. To the solution so obtained and stirred at room temp rature, a solution of 282 g (1.78 mole) of KMnO~
in 26 L of water was added dropwise over ~ hours. The mlxture was then stirred at room temperature for 1 hour and H202 (37%) was added dropwise until a colorless solution was obtained. 30 L of water were then added~ the mixture stirred for 1 hour at rv~ $~rature~and the cr~stalline precipitate was collected, washed with ~ x ~ L o~ water ana with 2 x 2 ~ of ethanol and dried over ~acuum at room temper-ature.
Yield: 297 g (76~) D (~5% C ~C12) ~ 75-9 Example 14 Preparation o~ BL-P2013 Free Acid ~s~ c ~ Cl ~ po4 ~ C ~ Cl ~ ~E3 ~ ~ r ~
d~ "C2~K~ ~--"'C02H

.

~ ~ 7~5~ ~ ~

To a mixture o~ 25 ml of ethyl acetate and 10 ml of water was added 800 mg (0.00261 mole) o~ BL-P2013 potassium salt. A~ter all o~ the solid had dissol~ed~
the mixture was treated dropwi5e with 50% aqueous phosphoric acid with vigorous shak~ng until no more material precipitated from the aqueous layer. The ethyl acetate layer wa~
separated, then washed wltA satuated sodium chloride solution and dried over anhydrous magnesium sul~ate, The drying agent was removed by filtration and washed with 10 ml of ethyl acetate, (m e wash solvent was combined with the original filtrate ) . I'Skellysol~re B" was then added to the eth~l acetate to the cloud point (approx. 10 ml). The mixture wa treated with 500 mg o~ actl~ated carbon ("Darko KB'I) and filtered. The ~iltrate was diluted with 15 ml of "Skellysol~e B", then seeded with crystals o~ BL-P2013 free acid. After approx. ~ hours at room temperature, the crystalline preclpitate of free acid was collected and dried in acuo ~15 min) over P~05 to obtain 3 3 mg (46%) m.~. slow deco~p. over 100.
Anal. ~.Calcd._ for C8~10ClN ~ S: C, 35.89; H, 3,~7; ~5~23, Cl, 13.25 Found: C, 35.88, H, 3.91; N, 5.41; Cl~ 13.52 This product was ~ound to be unstable when stored at 23C. for se~en da~s.

*Trademark ~l~ll7~ 8 . .
~e~
6a-Bromopenicillanic Acid Sul~oxide _ O
B ~ ( ~5C ~ N~CH2)2 ~

MW 800.6~ 96.14 ... _ . .. .. _ ,. , , , _ To 3 1 o~ methylene chloride was added ~00 g (0.75 mole) o~ 6a-bromopenicillanic acid N,NI-dibenzyl-ethylenediamine salt and this suspension was cooled to 5. Then o~er a 15 min~ perlod~ with good stirring, 130 ml of conc. HCl was added dropwise. The slurry was stlrred at 5 for 2 hours, It was then filtered through a ( "Celite" ) pad of diatomaceous earth and the cal~e was washed wlth ~ x 250 ml of methylene chlor~de.
The combined methylene chlorlde solutions were washed with 2 x 500 ml H20 and dried over sodium sulfate for 15 min. The sodium sulfate was removed by flltration and the ~iltrate ev~porated under reduced pressure to approx.
750 ml.
This solution wa~ cooled to 5 and, with vigorous st~rring3 1~0 ml of 40% peracetic acid was added dropwise such that the temperature was ma~ntained at 5 to 12.
The additlon was quike exothermic. At the end of the addition~
the slurry was stirred ~t 5 for Z hour~ and the product collected by filtration a~d washed wlth 100 ml o~ cold H20 (5) and 100 ml of cold methylene chloride (5). There was obtained 12~ g (57%) o~ 6a-bromopenicill~nic acid sulfoxide, m.p. 12go. The ir and nmr ~pectra were conslstent ~or the desired product.
Anal~ Calcd. ior C8HlOBrN04S C, ~2.44, H~ 3.40; N~ 4-7~-. . . ~ .
~und: C~ 32 .~; Hg ~.~5; N, 4.71;

H20~ 2.18 ,.. ~, . ........ ~...... ,. ,....... ,. ,~ , .

- ~42--~L7.5~

Potassium 6a 33romopenlcillanate Sulfoxide O O
Br~" ~ co2~ C0zK

MW 3~4.24 To 3 1 of acetone was added 126 g (0.43 mole) of 6a-bromopenicillanic acid sulfoxide and 162 ml o~ 50%
by weight potassium 2-ethylhexanoic acid in n-butanol.
After stirring 1 hour at 22, the product was collected by ~iltration~ washed with 2 x 250 ml of acetone and dried.
There was obtained 127 g (90%) of potassium 6a-bromo-penicillanate sulfoxide, m.p. 185. The ir and nmr spectra were consistent ~or the desired structure.
~nal. Calcd. for C~BrKN045: C, 28.75~ H, 2.71; N, 4.19.
Found: C, 29.0~; Hg 2.78, N, ~.04.

_A~_.
~IL7~

p-Nitrobenzyl 6a-gromo~enicillanate Sulfoxide __ O O

~C02K ~P~ 2 H~No2 MW 431.~8 To 1 1 of N,N-dimethylacetamide was added 145 g (0.43 mole) pota sium 6a-bromopenlc~llanate sul~o~.ide, and, with stirris~g, there was th~n added 115 g (0.5~5 mole) of p-nitrobenzyl bramide at 22. The mixture was stirred at 22 for 20 hours~
~; The re~ction mixture was poured into S 1 of H20 and extracted with 3 x 1500 ml of ethyl acetate. The combined ethyl acetate extracts were washed with 2 ~ 500 ml of 5~ aqueous sodium bicarbonate solution and dried over sodium sulfate for 1/2 hour. me sodium sulfate was filtered of~ and the filtrat~ evaporated under reduced pressure to a residue to which 1 1 of diethyl ether w~s added causing the product to c~ystallize. m e cr~stals were collected by filtration, washed with 2 x 100 ml of diethyl ether and dried to yield 162 g ~87~) o~ p-nitro-benzyl 6a-bromopenicillanate sul~oxlde, m.p. 111. The lr and nmr spectra were consistent ~or the desired structure.
Anal. Calcd. for C H 6BrN O~S: C, 41.78; H, ~.51; N, 6.50.

Found: C, 41.66; H, 3.45; N~ 6.85;
H209 o.6g.

,, . ., , . .. .. . , . . . ,, . . .. ,. . ~ , -. - , . ~ . ~

~:~7~30~1 3-carboxylate '~ ~ Br f ~ CB2Cl CH ~ N2 ~ ~ N0 MW 449.71 To 1 1 of p-dioxane was added to 70 g (0.16 mole) of p-nitrobenzyl. 6a~bromopenicillanate sulfoxide ~ollowed by 21~2 ml ~0.10 mole) of benzoyl chloride and 21.8 ml (0,19 mole) of quinoline. The reactlon mixture was refluxed ~or 4 hours and then cooled to 22, poured lnto 2500 ml of H20 and extracted into 3 x 800 ml of ethyl acetate. The c~mbined ethyl acetate extracts were washed with ~00 ml of 5% aqueous ~odium bicarbonate soiution, ~00 ml o~ 5% aqueous phosphoric acla-an~ ~0 ml of ~ 0. The ethyl acetate solution was dried over sodium sul~ate for 1/2 hour and the sodium sulfate was removed by ~iltration. The f~ ltrate was evapor~ted under reduced pressure to a r~sidue which was redlssolved in 1 1. of ethyl acetate and again evaporated under reduced pre~sure to a res due. Then 1 1. of diethyl ether wa~
added and the product collected by ~iltration to yield 41 g (57~ o~ p-nitrobenzyl 6a~bromo-2~-chlorometh~1-2-methylpen~m-3-carboxylate, m.p. 132~. The ir and nmr ~pectra were consi~tent for the desired ~tructure.
_ ~or C15H14BrClN205S: C, 40. o6; H, 3.14; N 6 23 Found: C, 40.62~ H, 3.11; N~ 6.13.

~ 5-~:gL75~

p-Nitrobenzyl 6a-Bromo-2~-chlorcmeth~ 2-methylper~am-3-carboxylate Sul~oxide O
Br ~ ~ 2Cl r, ~ CX2C1 2C ~ N0 C2C ~ No2 - MW 465.71 ._ . .. . , _ _ .
To 120~ ml o~ methylene chloride was added 51 g (0.11 mole) of p-nitrobenzyl 6~-bromo-2~-chloromethyl-2-methlypenam-3-carboxylate ~ollowed by 2~ g (O.12 mole) of m-chloro~roxybenzoic acid. The solution was st~rred ~t 22 for 2 hours and evaporated under reduced pressure to a wet residue. The residue W&~ stirred with 4 1 of diethyl ether for 1 hour and allowed to stand at 10 ~or 20 hours.
The product crystallized out and was collected by ~iltratio~, washed with 2 x 200 ml of diethyl ether and dried~ yielding ~9 g p-nitrobenzyl 6~-bromo-2~-chloromethyl-2-methylpenam-3-carbox~late sul~oxide (75~)~ m.p. 1~2. m e ir and ~mr ~pectra were consistent for the desired structure.
An~l Calcd. ~or C15H14BrClN206S: , ~
Found: C, 3~.98, H, ~.04; N, 5.84;
H20, 0.35~

~7~8 Potassium 2~h oromethyl-2-meth,yl~enam-3-carboxylate .
O O

'CN3 0 CH ~ No2 ~CO~K

_ _ MW 305.77 To 600 ml o~ H 0 was added 8 g of ~0% Pd on "Celite" and 16 g (0.19 mole) of sodium bi~arbonate.
en 32 g ~o.69 mole) o~ p-nitrobenz~l 6a-bromo-2~-chloromethyl-2-methylpenam-3-carbo~ylate sul~oxid~ was dissolved in ~00 ml of eth~l acetate and added to the aqueous slurry, The mixture was hydrogenated on a Paar apparatus at 50 p~s.i. at 22 for 4 hours. The slurry was filtered through a thin "Celite" pad on a s~ntered gla~s funnel, the pad was washed with 2 x 50 ml H20 and the agueous layer of the ~ombined filtrate and washings was separated~ The aqueous layer was washed with 200 ml o~ dieth~l ether, then was cooled to 5 and, with st~rring, a solution o~ 12 g. (.o76 mole) of KMhO4 in 200 ml of ~ 0 was added dropwiRe over a 1/2 hour period~ keepin~ the pH
betw~en 7..5 and 8.o by the addltlon o~ 40% ~PO~. ~hen the pink color persi~ted ~or 5 mi.nutes, no more KMnO4 solution was added. The reactlon mi~ture was stirred with a small amaln~ (approx~ 50 mg) of sodium bisulfite for 1/2 hour, and then the ~lurry was filtered through a "Celite"
pad. The pad was washed with 2 x 50 ml of ~ 0. The . . . . .. .... ... . .

_ . .
~ ~ 76~

combined filtrate and washlngs were layered with 500 ml of ethyl acetate and, with stirring, the pH was adjusted to 1.5 by the addltion o~ 2 N HCl. The layers were separated and the aqueous layer was saturated wlth sodium sulfate It was reextracted with 2 x 400 ml o~ ethyl acetate and the combined ethyl acetate extr~cts were dried o~es sodlum sulfate ~or 1/2 hour at 5. The sodlum sulfate was removed by filtration and the filtrate e~aporated under reduced pressure to a residue. That residue was dissolved in 1~0 ml of acetone and 160 ml o~ diethyl ether and 50%
b~ weight of potassium 2-~thylhe~anoate in n-butanol was added until the solution was n~utral to molst pH paper.
The potassium salt o~ BL-P201~ crystallized out, was collected b~ filtration, washed with diethyl ether and dried. ~ield 16 g potassium 2~-chloromethyl-2 methylpenam-3-carboxylate sul~one (BL-P2013) (76%), m,p. 202. The ir and nmr spectra were cons~stent ~or the desired structure.
Ar. ~.- Calcd. ~or C8HgCIKN05S: G~ ~1.42; H~ 2.97; N, 4.58-Found: C~ 31.18; ~3 2.98~ N, 4.51~ . ~~ - H~0, 0.~3.

.

-~8~ 7 Example 16_ Pivaloyloxymet~2~ -Chloromethyl--2-me kh2 lpenam- ~-carbox~late Sulfone (BL-P2024~
O O ~ O
~S/1 CH2C1 ~s~H2Cl F~2K F~ C02C~20COC(C}~

_ _ _ ._ _ MW 381.83 To a stirred suspension of 14.6 g (0.0487 mole) of potassium 2~-ehlorometh~1-2-methylpenam-3~!carbox;ylate sulfone (BL-P2013) in 200 ml of acetone was added 4 ml of a 10% a~ueous solutio~ of sodium iodide and the ~xture was brought to re~lux on the ste&m bath, To thi~ re~lux-ing æuspension was 2dded 14.8 ml. (0.1 mole) of red~stilled chloromethyl pi~alate (bp 34C at 7 mm Hg) all ~t o~ce.
The mixture was stirred at reflux for three hours and then cooled to room temperature (22 C). The crystalline solids were collected b~ filtration, washed with 3 x 30 ml of acekone and the combined ~iltrates were evaporated to a~
oil under reduced pressure at ~22 C. The oil ~as then taken up in 500 ml o~ ethyl acetate and washed once with water (200 ml) and once wlth ~aturated Na2S0~ solution (200 ml). The solution was then dried brie M y over Na2S04 while being stirred with 2 g o~ decolorizing carbon with cooling (ice bath). After 20 min. the mixture was ~iltered through a "Celite" pad and the pad ~ashed wîth 4 x 100 ml o~ ethyl acetate. The combined ~iltrates were concentrated under reduced pressure at 2~ C to ~n oil~ me oil was then further concentrated ~t about 22 C and ~l mm ~g to remo~e most o~ the residual chloromethyl pivalate. The xemainin4 oil was then tritur~ted twice with 50 m:L portlons of n-pent~ne and then le~t o~er the weekend at about 10 C

';

~7~

under n-pentane. The resulting solid crystalline mass was then broken up to a powder under 40 ml of a 4:1 mixture of diethyl ether-n-pentans. The product was the~ collected by ~iltratlon, washed with diethyl ether-n-pentane (1:1~ then n pentane and air dried. After dry~ng under high vacuum for fo~r hours oYer P205 there was obtained 13937 g pivaloylo~ymethyl 2~-chlorome~hyl-2-methylpenam-~-carboxylate.sul~one (BL-P2024) about (75%), m p 9~ g5 C

~50-~7~

Purification of BL-P2024 .

~ pproxim~tely 3 g. of crude BL-P20?4 (obta1.ned as described abo~e) wa~ dlssol~ed in 5 ml. of ethyl acetate, placed on a 4.5 x 40 cm. col~n of ~ilica gel (M~llinckrodt CC-7), and eluted with 4:1 ~/v C ~ C12-ethyl acetate. The ~ractions containing a. single spot at RfO. 84 (TLC on sllica eel plates with 4:1 CH2C12-ethyl acetate, I2 detection) were combined and concentra-ted under reduced pressure to 1.38 g. o~ a crystalline solid. A portion o~ this material (900 mg.) was dis-sol~ed ~n 5 ml. of ethyl acetate; the resulting solution was filtered, diluted almost to the cloud point with 'petroleum ether ('ISkellysolve B") and then ~tored at room te~perature for three days. m e crystals which ~ormed were collected by filtration, washed w~th petroleum ether and dried to give 560 mg., m.p. 100-101., of purified BI~P2024.
Anal. Calcd. for C14~ ~Cl~O S: C, 44.03; E, 5.27; N, ~.67 Found: C, ~4011; H, 5~08; N, 3.85.
All temperatures in this applicat~on are given ln degrees Centigrade.

.

~ ~.7~

Example 17 Preparation o~ BL-P2013 ~mmonium Salt 1. m e ~ree acid of BL-P2013 (250 mg.) dissolved in 20 ml. of acetone-methanol (1:1 by volume) was ~ ered to get a clear solution.
2. Anhydrous ammonium solution was prepared by adding 1 ml.
of ammonium hydroxide (~0%, reagent grade~ to lO m~. o~
acetone-methanol (1:1 by volume) solvent and then 1 gmO
of anhydrous magnesium sul~ate was added to that solution with mild agitation and the mixture was Piltered thrQugh a ~ilter p~per; the filtrate was designated "anhydrous a~monium solution.'i 3. To the filtrate of Procedure 1" approximately 2 ml. of "anhydrous u~monlum solution'l w~s gradually s.dded snd mixed w~ll .
4. A 100 ml. portiQn of diethyl ether was mixed with the mixture from Procedure ~ tQ pr~cipitate the ~Lmmonium salt of BL~P2013.
5. m e white ammonium salt w~s isolated from the solvent and washed with 2 portions of 50 ml. each o~ diethyl ether.
6. me isolated powder was dried at ~5 C vacuum oven for overrlight .
7. Analytical data were as ~ollows:

Calculated % C 33.7; H 4.6, N 9.8, Found C 3~;.66; H 4.63; N ~O.12; drg by KF
Micro~copic Examinatlon: crystalline sub~tance .. . . . . . . .. . . . . . . ... .. . . .. .. .. .. .. .

~52--E~cample 18 1. Dissolve 50 mgO o~ the free acld o~ BL-P201~ in 4 ml.
of acetone-methanol (1:1 by volume) mi~ture. Filter to get a clear solutionO
2. Prepare sodi~m 2-ethylhexanoate solution by dissolving 40 mg. of sodium 2-ethylhexanoate in 10 ml. of acetone-methanol (1:1 by volume) mixture.
3. To the filtrate of Procedure 1~ add the I0 ml. solution of Procedure 2 and mix well.
4. A 10 ml. port~on o~ diethyl ether was mixed wlth the mixture ~r~m Procedure 3 to precipitate the sodium s~lt of B~-P2013.
5. ~he white salt was immersed in the dieth~l ether ~or 1-2 hours and th~n was isolated from the solvent and washed with 3 port~ons of 5 ml. each of diethyl e~her.
6. The isolated powder was dried at 30 C ~cuum o~en for overnight.

.

--53~

// CH2Cl Recryst~ C~12Cl _ ~ S~,. ~ _,~
I" CH3 H20-acetone r~ H3 0~ 3~ " C2K'H2 ~' t~~K~H2 B~P2015 (400 mg.) was dissolved in a minimum amount of acetorle~H20 (1:1) by ~rolume arld diluted with 10 ml. o~
acetone, filtered, ~hen dlluted with acetone to abaut 25 ml., scratched, and after 30 minutes the crystalline hydrate was colleeted by filtration, washed ~7ell wlth acetone9 air dried and then vacuum dried at <1 mm. H~;
overnight .
Yield 280 mg.
Anal. Calcd. ~or C8HgClNOSK~ E~0: C, 29.67; H, 3.39, N, 4.63;
Cl, lO.g4, H20, 5.55-Found: C, 29.32; H, 3.32; 1~, 4.44, Cl, 11.31; H20, ~. 90 .

- .. .. . . . . . .......... .

Example 20 NlN'-Dlbenz~lethylenediamine Salt~

BL-P2013 + 1/2 N,N'-Dibenzylethylenedi&mi~e diacetate Recr~st. ~ / ~ // C ~ C1 Acetone-ether / ~ ~ C ~ \

~S~ C 02H ~6H5CEI2 ~J

\ ~ 2 ~ o6 mg. (0.001 ml) of BL-P2013 was dissol~ed in 7 ml. H20 and added to a solution of 180 mg. (0,0005mol) of N~N' dibenzylethylenediam~ne diacetate in 7 ml. H20.
The m~xture was stirred and the salt crystallized and after stirring approximately 1~-15 minutes the salt was collected by filtration and air dried to yield N,N'-dibenzylethylene-diamine salt of BL-P201~ (300 mg)~ The material was re-crystallized by d~ssolving it in approximately 10 ml. of boiling acetone and diluting with ether to the cloud point.
260 mg. o~ air dried and vacuum dried material was obtainedn Anal. Calcd: C, 51.69; H7 5.42; N, 7.5~; Cl, 9.55.
~ ~ . ~
Found: C, 49.39; H, 5.49; N, 7,05~ Cl, 8.96;
H203 1.23 (KF).

.~7SI~

Ex~mple 21 Chlorometh~l Ester of BL-P201 . . .

H2Cl + ClCH2-O-SO~Cl ~' C~ ( CE~;CH2c~2cH2 ) 4~SO4 0~ ~COOK + :3 K~ICO~;
(35 ~7) 5 I C~E2C12 ~ H20 O O
~3 O ~C~-OCH2C

(316 . 17) .7 . _ ..

-Sh-~75l~

To a vigorously stirred mlxture of 15.25 g ~0.05 mol) of BL-P2013 (5)~ 1~ g. (0.15 mol) RHC03 and 1.7 g (0.005 mol) of tetr~butylammonium hyclrogen sulf~te (Aldrich Chem. Co,) in a mlxture of ~0 ~1~ water and 50 ml. CH2C12 there was added dropwise a solution of 9.5 g (0.0575 mol) of ClCH2-0-S02C1 in 40 ml. CH2C12. The temperature rose to 26 C, and after the addition (w~ch took about 15 minutes) the mixture wa8 stirred Qnother 30 minutes. Because the product crystalliæed out more C~2C12 (abou~ 400 ml.~ was added to obtain a solution. me separated CH2C12 layer and a 50 ml. C ~ C12 wash were com-bined, dried over MgS04 with ætirring and 2 g of dPcoloriz~
ing carbon ("Darco KB") was added. ~ter about 30 minutes the mixture wPs filtered, concentrated to about 50 ml. and isopropyl ~lcohol (150 ml.) was added. The rest of the CE2C12 was then removed under reduced pressure. The resulting crystalline precipitate was col ected by filtra-tion9 washed well:with isopropyl alcohol and air-dried.
After vacuum drying at l~ss than 1 mm. ~g there was obtained 8.5 g of chloromethyl 2~-chloromethgl-2-methylpenam-~-carboxylate sul~one (~). M.p. 116 (dec., darkens above 100C).
Anal. Calcd. for Cg ~ lC12N05S: Cg 3~.18, H~ 3.51; N, 4.43, Cl, 2~.4~.
Found: C, 34.16; ~I, 3,45, N, ~47;
, C1~ 22.46~ H20J 0~33 (~)-~Estim~ted purit~ I~ the gO-95~ range~

~ ~7t~

Iodomethyl E~ter o~ BI,P2013 O O O O
~ /~ ~ CH2cl Na I ~ //~ CH2Cl ~ c~ r~ ¦ //CH

O C-OCE2Cl C-OCH2I

To a stirred mixture of ~ g. (0.01~9 mol) of the chloromethyl ester of BL-P2013 (7) in 25 ml. acetone was added 3 g. (0.02 mol) of sodium iodide. The resulting slurry was ~tirred for 1~ hours and then cooied to about OC. Two drops o~ saturated aqueous KHC03 were added and the mixture ~as slowly diluted dropwise with water over ten minutes u~til 50 ml. had been added. The slurry underwent a sudden color change ~rom yellow to grey to purple to black and therefore the crystals were ~mmedlately:collected by ~iltration and washed with cold acetone-water (1:2) and then isopropyl alcohol (~ x 10 ml), then diethyl ether and ~inally n-pentane and air-dried to yield 5.5~ g. (91~ yield) oi the lodomethyl ester of BL-P2013 (8). M.p. 118 - 119C. w~th decompositiQn.
Purity estimated at about 90%.
.. . . .
6-C(R)-2-Amino-2-phenylacetamido]-3,~dimethyl-7-oxo-.4_ thia.-.l-azabicyclo~3 2.0]~pt~e-2-car~oA~l~xymethyl 2~-chloromethyl-2-a-methylpenam-~a-carboxylate sulfone (11) ., . . . . . . . . .. . . . . ~, . .. .. . . . . . .. . .. .. . .... ... ..... ..

~ o - ~
~L~7~

8~ - ~CH-CO-IIH~3 oo~
n3~, ~ N

C~ C~O g ~CH3 (A Dane salt of' ampicillin, see U,S.
Patent 39316,247) \ /

~CH--CO~ ~ 3 N ~ C=O
H~C--C ~ ~
H C~ f ) O O
~S~ CH2C~
C!~ I 2 O
~ .
... .

, . . .... ~ .. , ., . , .. .... . .... . ... " . . ... . . .
. ~

. -5g-L7b~

CH - Co - NH ~ C

MH2 'CcO

O
To a s~irred mixture cooled in an ice-bath of 5.~6 g. (0.01 mol) of the ind~cated Dane s&lt of ampicillin 2 (wh~ch was solvated wikh one molecule of isopropyl alcohol) in 60 ml. acetone there wa~ added 4.o8 g. (O.Ol ml) of the iodomethyl ester of BI,P2013 ~8) and the resulting nearly cle&r solution was stirred for five hours with the ice-bath removed a~ter 30 m~utes. Then most o~ the acetGne wa~ removed ~n vacuum on the roto-vap and the resulting concentrated ~olution W&S dissolred ~n 200 ml. cold ethyl acetate whlch waæ then washed wlth 2 x 50 ml. ice coid water and 2 x 100 ml. saturated aqueous Na2S04. The eth~l acetate solutlon was then dried over Na2S04, filtered and most of the ethyl acetate was removed ln ~acuo on the roto-vap.
m e residue was triturated with 2 x 200 ml. dry diethyl ether and the resultlng solids were collected by filtrat~on to g$~e 5.5 g. of 10 as a p~nklsh powder. This powder was ..

--~o--~7~

stirred in a mixture of 50 ml. water, 50 ml. n-butanol and 20 ml. ethyl acetate while 6N HCl was added dropwise to pH 2.5. men occasionally a drop or two of HCl was added to keep the pH at 2.2 - 2.5 over 45 minutes. When the pH no longer drifted upward there was added to this mixture 100 ml. diethyl ether with good stirring. The aqueous phase was separated and combined with a second 25 ml. H20 extract of the organic layer~ The aqueous solution was extracted once with 50 ml. diethyl ether and the ether was discarded.

The aqueous lay2r was then stirred vigorously under a layer of 100 ml. 2-butanone (methyl ethyl ketone) while Na2S04 was added to saturate the aqueous layer. The 2-but~none layer was separated, drled over Na2S04 ~or ~0 minutes in an ice-bath9 filtered and concentrated in vacuo to near dryness. The residual oil was triturated to a solid with n-butanol, washed well with ether, then n-pentane, air-dried and then vacuum-dried over P205 at ~1 mm Hg pressure to yield 1.6 g. 6-[(R)-2-amino-2-phenylace-tamido]-3,~-dimethyl-7-oxo-4-thia-1-azabicyclo~.2.0]heptane-2-carbonylmethyl 2~-chloromethyl-2-a-methylpenam-3a-carboxylate sulfone (11) in crude form. The IR and nmr spectra were consistent with structure 11 but not with high purity.
This solid product waæ estimated to contain at least 40% and perhaps as much as 80% 6-~(R)-2-amino-2 phe~ly~ace-tamido]3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[~.2.0]-heptane-2-carbonyloxymethyl 2~-chloromethyl-2-a-methyl-penam-~a-caxboxyl~te sul~one.

.

S13~8 Example 2?

This procedure simplifies production of BL P2013 by eliminating the previou3 use of catalytic reduction.

O .
Br ~

~= + CEI2CC13 + Dicyclohexylcarbodilmi de Pyridine CH2Cl~
O
B~ 2 ,~N

O

(See page 6~5~ of Cephalosporins and Penicillins, edited b~
Edwin H. Flynn, Academic Press, New York~ 1972) 6a-.Bromopenicillan~ c acid sul~oxide (1) (30 g., 0.1 mol) was dis~olved :Ln 1 1. dry CH~C12 Pollowed b;!jr the addition of 16.2 ml. (0.2 mol) pyridine ~nd 29.8 g.
(0.2 mol) trichloroethanol. Then 20 g. (0.1 mol) of dicyclohexylcarbodiimide was added and the mixture was stlrred at 22 for 16 hours. D:lcyclohe:cylure~ began . -62- . ..
~7~

to precipitate out 3 at the end it was removed by ~iltration.
me flltrate was washed with 200 ml. of 5% aqueous sodium bicarbonate, 200 ml. o~ 10~ phosphoric acid and 100 ml.
of saturated ~queous sodium sulfate. The organic phase was dried over sodlum sul~ate at 5C. for ~0 minutes, filtered and evaporated to an oil. Diethyl e~her was added and, with scratching, the product 2 crystallized out (27 g., 63~ yield).

2 * ~ 8 CCl ~ Qulnoline ",F~CN2Cl C- OCX2CC~

Compound 2 (~6.5 g., 0.062 mol) was d~ssolved in 500 ml. p~ oxane and there was added 8.5 ml. (o.078 mole) benzoyl chloride a~d 8.75 ml. (o.or8 mole) quinoline. The solution was re~luxed for ~our hours and then poured into 1100 ml. water and the product ~ was extracted lnto 2 x 400 ml. ethyl acetate. m e ethyl acetate extracts were combined, successively washed with 200 ml. 5~
aqueous sodium bicarbonate, 200 ml. 5~ phosphoric ac~d and 200 ml. saturated aqueous sodium æul~ate~ dried over sodium sulfate ~t 5C. ~or thirty minutes and evaporated to an oil (~) which was used l~&S iS~I for the next reaction.

... ..

~ ~ 7 KMnOl~,~202 in glacial ~cetic aeid : ~
\ /
: O O
: Br \\ ~ ~CH2Cl N____J"
: 0 C~OCH2CC
~ O
:
Compound ~ obtained in the previous 3tep was dissolYed in l 1. glacial acetic a~ld and, with st~rring at 22C., a saturated aqueous solut~on o~ KMnO4 wa~
: added dropwise until a plnk color peræ~ted (that is, a drop placed o~ a piece of ~ilter paper ga~e a pink coloration). men with cooling ~% ~22 was added dropwise until ~ clear solution was obtained; some white precipitate w~s present. The olution was poured into 2.5 1. water and the produ~t 4 was extracted into 3 x 500 ml. ethyl acetate. Ihe ethyl acetate was washed with 5% aqueou~ sodium bicarbonate until neutral (that is~ no more bubbling upon addition)~ dried o~er sodium 3ulrate and evaporated to leave 4 as the residue.

--~4--S~

Xt was le~t at 10C. for one day and then triturated with "Skellysolve B" to yield 9.1 g. sol~d 4. The yield was 28~ o~ theor~ for step~ 2 and ~ combined.

St~p 4 + ~ Zn in Acetic Acid KEH

/ CH Cl ~ 3 5 (BL~P2013 ) o COOK

(See U. S. Patent 4,16~,497) Zinc dust (3.75 g.) was slurried in ~ ml. glaclal acetic acid and cooled to 5C. To this ~ ture there was added a solution of 4 (~ g.; 0.0057 mole) in 15 ml, d~methylformamide and the re~ulting slurry was stirred at 5 for 2.5 ho~rs.

The zinc was then ~iltered of~ a~d the pale yellow solution was poured into 80 ml. of 5% aqueous hydrochloric acid. That mixture was extracted ~lth 3 x 25 ml. ethyl ~cetate, The comb~ned ethy~ acetate extracts were extracted with ~ x 20 ml. of 5~ aqueou~ sodium blcarbonate, saving the ethyl acetate phase after separationO

-6~-S~

m e bic~rbon~te extracts were combined~ placed under a layer o~ ethyl acetate~ ad~usted to pH 1.5 by the addition o~ 2N HCl and saturated with sodium sul~ate.
The eth~l acetate was separated and the aqueous phase was extracted with 2 x ~0 ml. ethyl acetate.

All o~ the ethyl acetate phases above were combined, dr~ed over sodium sul~ate and e~aporated to an oil (which was the ~ree acid form o~ BL-P2013) which was dissolved in about 20 ml. acetone to which 20 ml. diethyl ether was then added. m en 50% potassium 2-ethylhe~anoate ~KEH) in dr~ n butanol was added to neutrality. The product 5 (~I-P201~) crystalli~ed out. After stirring 0.5 hour at 22 it was collected by flltration to yield 650 mgm. of 5 (37% yield).

A 50 mgm. sample o~ ~ was dissolved in 0.~ ml.
water and 20 m~m~ N,N'-dibenzylethylenediamine (DBED) diacetate was added. The DBED salt o~ 5 crystallized out, was collected by filtration, washed with water and dried over P205 under vacuum to yield N,N'-dibenzylethylenediamine 2~-chloromethyl-2-methylpenam-3-carboxylate sul~one (DBED
salt of free acid 5).

Another ~ample o~ 5 (450 mgm.) was dissolved ~n 3 ml. water to which was added a ~olution of 270 mgm. DBED
di~cetate ln ~ ml. H20. With scratching the DBED salt o~
crystallized out (430 mgm.). Recrystallization ~rom about 5 ml. boil~ng acetone yielded 270 mgm.

--.
.

~.~.7~

6-~(R)-2-~mino-2-p-hydroxyphenylacet~mido~-3,~-dim~thyl-7-oxo-4-thia-1-azabicyclo[~.2.0]heptane-2-carbonyloxymethyl 2~-chloromethyl-2-a-methylpenam-~a-carboxylate sul~one having the ~ormula _ _ HO ~ I ~ 3 O O `
~S~ CH2 . , ~ ~
i~ produced by substituting the correspond~ng Dane salt of amoxiciilin ~or the amplcillin used in the procedure of Example 21.

BIOLOGICAL DATA

me product of Example 1, C:ompou:nd 5 y ha~ing the structure ,~o r~C~3 ~ N "~
tJ CQ2 R

will be referrea to below a~ BL-P2013~

Alt~iough at bes~ a Yery ~eak antibact~rial agent itself, BL-P2013 inhibits i3-lactamas~s asld prot~cted ~eforanide and amoxicillin from ae~truction by ~-lactamase-produc:ing }:~acteria in ~itro arld in Vi~To when used in co~bination with those t~wo agent~.

.

Table 1 Antibacterial Activity of the New Sulfnne . ~ _ . ~
MIC ~mcg~ml ) .
Organism : BI.-P2013 Ampicillin _ . _ ~ .
~ ~ _ ~ , S. pneumoniaaA-95B5 16 0 . 004 . pyogenes A- 9 6 0 4 _ ~_ s . aureusA- 9 53 7 _ >12 5 0 ~,1 6 . _ _ _ . . .
~ aureus ~
50% seru~nA-953~ >125 O. 06 S. ureus Pen- _ Res A-9606 >12S ~125 S. aureus Me~h- _ :: Res A15097 ~125 125 ~ ~ . .... . . ~
S. faecalisA20688 >125 0.13 : ~ E. co}i A15119 >125 ~ 1 E. coli A203~1-1 ~
K. pneu~noniae A15130 >125 125 _ _ ~ _ K. pneumonia~ ~20468 >I25 1 ~125 P. mirabii~s A-9900 ~125 0.,13 . ,_, . .. _ . _ ...... ~ ~ . . . . , P. vulgaris A21559 >125 125 ~ --I
P. morganii A15153 ~125 >125 P. rettgervi v - -i~21iO--- - ~12 5 ~ ~ ~~ 4 _ . S.. marcescens ~20019 ~125 1~:
E. cloacae A-9659 :~125 - ~
. . _ , ,, .. ~ ~ . ~_ I~ _ _ . . - - . - I E. cloacae A-9656 ~125 ~125 - . - ... _ _ .. .. ~ ..
P. aeruginosaA 9843A ~125 ~125 P. aeruginosaA21Z13 >125 ~ ~~ >125~ = .

-~75~

Table 2 -Ar~ti-Bacteroides Activi~y of Cefora~ide and Amoxicillin Alone and in Combination with BL-P2013 _ Beta~ [~ T~
Organism lact Ce~o= Cefora~ide BL pATnoxicillin A~ioxi-r _ r amil 8 C _ . fragilis .
A2191 ~ 63 2 > 125 2 8 ~22053 + 32 4 63 2 8 A2202 + 32 2 32 2 4 ~2187 + 3~ 4 63 2 8 A2253 +~125 32 125 16 >125 A2269 ~ 63 8 63 2 8 A2269 ~ 63 4 63 2 ~6 A2269 ~125 16 63 2 16 A2259 . +>125_16_ 32 4 125 A2269 l>125 _32 63 8 ~125 : ~2~5~ ~>1~5 _32 32 32 >125 A2253 +~125 32 125 32 ~125 P~2279 ~~125 8 32 4 12 A2279 ~ 32 4 32 2 8 1~279 + 32 4 32 2 8 ~2279 +6 :1 4 32 4 16 A2~79 I ~3 ~ 63 2 16 A2279 + 32 4 63 2 8 ES. theta- .
iot:aomic-ron A2227 ~ 125 4 63 2 16 A2227 l 125 8 63 4 16 _ _ __ _ _ .

-' :

,:

Table 2 - cont'd.

Anti-Bacteroides Acti~ity o~ Ceforanide and Amoxicillin Alone and in Co~bina~io~ with BL-P201~
. _ ~_ Beta-Organism lact- Cefo- Ceforanide BL-p Amoxicillin Am-oxi~
. ~mase (lPl)13 2013 BL~ .3cilli~
~ _ . . .~
Bacteroides _ pecies A2G934 ~ 32 4 32 2 8 A21959 + 63 ~ 32 2 16 A20929 + 63 4 3~ 2 16 A~1954 ~: 63 16 63 2 16 . A2093 ~ 6~ 3 _4_ 8 A2093t ~ 125 8 125 4 32 ~ A2093] ~ 63 4 32 2 1~
. A20927-~ _ 0.S 1 63 0~13 ~.1 A2093' _ --- good synergism --- marginal synergism . ~,, , , , . _ . .
*Mini}num inhibitory concentration (MIC) determined by the agar dilu tion method using SOX dilutions o~ 2 4 hour cultures as inocula dispensed by the Steer ' s inoculator,. A~csay medium composed o~ Brucella Agar plu5 551~ laked sheep blood and 10 mcg/ml vitamin K.

- , Tabl e 3 Therapeutic Efficac~_of Amoxicillin in Combination with BL-P20l3 in Mice E:xPerimentallY Infected wit}l a Beta-_ Lac:ta3nase Strain of StaE~hy~c:o~cus aureus ~hallenge PD5~/Treatment~mg~kg) ~
organism ~rganisms) ~o~icill~ 1 BL-P2 013 B) A~B ~ l) S~ aureus S x lO~ ~800 ~80Q >50 ~200 6.3 44 A-9606 5 x lO~ >800 >800 >50 >200 l9 77 7 ~ I D > 8 00 -- ~ 50 _ 9 . 6 --:: :
Treatment sc:hedule: I)rugs administered a and 2 hours po~t-ection., " :

: ' _7~ 75~

. ~... ~.. ~ _.... _ o~ ~o ~o o~ ~o ~o ~a ~c) ~ ~U ~ D 0 ~n 0 ~ ~ O~ O~ G~
o l ., ., ., ., ., o ~ ~
_~ _.
, _ q~ ~ ~ ~ , o ~, o o _~ ~ T1 ") U~ ~ O
. ~ .
_ _ _ _.
.
~J O ~D ~ 1~ N ~
I . . I . . .
~4 ~ o ~ ~ v v O
: ~ ~ O ~ In ~ U~
O~ ~ h . .
E~ _~ ~ ~ 3 Il~
X~ ~U ~ V Q~
_I :3E ~ _I ~ ~ 1 t~ 1~
~ O ~ o E3 o o ~ It~ :~
~ C: ~1 ~ V O
~ O ~ ~ ~ o~ _l r` ~ O
-=t ~ 0 ~ ~1t`~ O~
a)J~ ~ ~ ~ ~
,1_I ~ ~ ~ a~ ~r J~
~ ~D O Ql ~ ~ 0 ~ .
: ~~a rl O O .~1 7 ~I N ~ ~1 4 ~ :: ~ _1 ~1 ~ ~n ~; ~n ~ u~ ao P~ ~ ~ ~ ~ ~
_, ,. . .__ . ., m ~ . . ~
C:~ O o o O O
O ~ O b~ O O O o o o a~
VJ ~¢ a ~ ~ ~ _I N a~
~ ._ _ . ~ o~
.
Q 1~1 ~1 q) 3=~ I ~ ~

. ' :

.

.
.

c ~ ê . -~ - .8 _ - ....
O t` ~ ~a ~
e ~ s~ r ~,~ O

C Iq u7t~ ~ o R. 0 O r~î ~ ~ ~ ~ U~ ~ ~ O
~9 ~ O ~~` N . n ~ ill ~C ~ ~ Æ ~ ~ ~ Q tJ
e~ 8 s~ ~ ~ ~ ~ ~t ~4 e rl N 19 dl 1~ ~ U V ~ 10 U9N~Dr U 0 o O 0~ 0 ~ ~ X ~ ~ 0 Il) ul ~C ~ I~ D
~ v ~ u~ ~ ~ ~ ol ~
~ ~5 _~ " ~ --I U
~ .le ~c ~ 0~q ~ #~ O O O ~ ~
8~, ~ ~ Y~o o ~ ~ o _..... ~ Y
h e o ~ ~ ~ ~ h ~ o ~ ~ ~ ~ ~ 3~ ~
;

5 !3C~8 ~, ~1 ~ ~ o o o o ~ o ~> o c~ o ~
o , _. o C o o ~` o Cl o o o s~
c7 m ~ N N ~ I'il N
_ ~
~ ~ O ,D Il~ ~1 V ~ ~1 _ N
~1 _ N ~1 r~ ~' NO ~~1 ~r e~ o ~ ~ o q~ ~ ~ ~D ~ ~ _~
O o ~ ~ _1 ~ ~ 7 W
m c _ ~ _. ~, ,", O ~ ~
~, ~ O ~ ~ _, 3ul ~ Cl. ~ r` cr.
E ~1 ~ t'~ N ~1 It~
O C 1:1 N ~r ~ o ~ ~r o rl a~
d V ~ ~ r- ~ ~ I
U ~ ~ ~1 c~n ~ rl O
.n ~. ~ ~ u~
Ll U ~1 A ~ ~Y
~O 1~ E~ ~ ~ o 4 1 c ~ a ~ .~ ~ O ~ ~
~ i o . _ _ X C X O O O O O O O O O O
3~ 3 L~ A ~ A A ~ ~ ~ ~ ~
. _ ~.__.__ , c 1~1 c~ It~ In In ~ Ul 0 ~ ~ O O C~ O O O O O O O O
V C O C ~ ~ 1 I ~ X X ~ X X X X ~ ~ X :~e t4 ~ ~ ZO O t'` t~ O IrlItl ~ I`
_ _ _ ~
~ ~: 1~ ~ ~ ~
_1 ~ ~ ~ 5~ 6r ~: ~ o ~ o o ~ o ~ ~ U ~ U
o ~q 2 ~o ;! ui ~e ~i .

_75~
., ,._ ~. _ .. __ ol ~ o o o o C~

~ W o ~ C
~ e ~ -I ~ ~
~ h ~ ~ . O
0~ V ~ ~ V
~a ~ . u7 ~ o ~1 1~1 rl o t) _~ ~ ~ ~
E~ O ~ ~ o 0 co ~ .. __ m ' ~_i o E ~1 O ~ A A ~C
_ E ~ c~
O ~'q, '_ ~ ~
u ~s: ~n In In U~
o o o ~ C-I O c: ,~ ~ ~1 _I H~1 1~
~1~1 01 X X X
W ~~C O
~ 5 = . .~ ~ c o o ~i ~ t s:: ~o ul .~ 3 ~: a) tl t~ 5 r~
o .c ~o ~

_ _ _ ~ ~8 ~o~
o 3C
o C ,1 ~
~ ~ ~ ~r I~ _~ .
_l u~ O
c ~ E cr. ~ ~r u7 r~ ~3 o :~ ~ v ~ ~ O
~ c ~ ~
~ O F _I ~ U~ ~ W 1` Ul 1` C:l .1 1~ 0 ~ ~1 ~_~ a~ 0 ~ U Ul ~r . o C O ~ ~I _~ ~ . . 1 ~1 E~ X _1 ~ 1 C ~
I; C O E~ ~ ~ OUl N .SJ ~ 3 aU a ~ ~

t_ O O i~ ~ _~ O O O O O ~ I r~ _~
~ ~ ~ 3 Q ~ o o o o o o ~ o :1 .CI ~ ~1 . ~ ~ ~ A ~ ~ ~1 0 ~ C q~ _ __ ~ 3 >~
~ O O .1:1 g 5 S: I 1) U Vd~ O a~ O Cl U N ~
o ~ ~_~ ~ :~ o u~ o u~ 2 ~E ~ X
C ~ C~: C ~ ~D
'4 ~ ~q a :~ ~ ~ -~
C . .. - ~ o O t ;~
~ ~ ~ ~ 0 .
o C ~ ~ ~CO Ul U~ -~ sl ~ o c o o o oo o ~ ~ s 0 ~ ~ ~ ~ 1 ~
z o x ~ ~ xx x o ~O o ~_ u~ w ~
C~ ~ ~ . _._ _ . . ~ OC
~ ~ ~ ~ ~ U ~q rt ~ ~ ~ t) i~ C
_~ ~ ~ h o~
P o al~ c ~ 1 1'` o u7 0 u~ O ul O u~ O ~ C
X U~ C I~ ~ o ~ U U 1` ~ ~o ~
a~ t ~ ~.1 , ~ N ~ ~ hi~
o D~ ~ ~

_-- O -- n----o ~,~ a: ~ ~
u~ ~o ~ o ~ ~oo in c~i ~
o o o o o _, _ _ - ~ _ _ û~ _, ~ .~
_i N i :S~
O ~1 1 i~ I ~1 1 t- I
N i ~0 i~ i ~_10 Cli O O ~i V O r1 ._ _ t- ~i _~
~ _ e~ CU N ~i tl~
O ~ f~ ~ ~ l ~ I ~ I
~e o ~ ~D i--~.~ ~ ... .
~ i;~ ~ri C ~-i O ~-i O ~i N
~ ~ ~ ~i V _ _ V _ _ ~i ¢ _ ~ ~ _ ~ _ a:l ~1 ~ ~ ~ ~ ~ ~D ~
0 0 ~i U~i~ ~ O~ ~CI ~:t ~ ~ ~ ~ ~ C)~ ~ r~ r1CO ~ ~
lD ~ S~ O Ul O ~i ~ ~ ~ ~ ~i O ~D O ~ _ _--Il) N ,a rl ~ ~ _ _ _ r~l ~:4 i- t~i~ ~_t ~O ~:U ~ ~D
~: _ ~ I~ i ~ ~ Y
a:~ ~ ~ ~ ~ ~ o~ I ~ ~ ~o i u~ I ~u 6~ ~i i~ N~i ~ ,~Cr~ ~i n'-~ ~D o ~ _~
O E-~ N a _ _ N _ I O
~0 _ __ -......... ~
c~ ~ ~D t- ~ ~ ~ ~ O
C~i 3 . 3i~ ~ N ~ U~ t 0~
~~1 1 CU I ~1~ I CU I ~ I a- ~
.: ~ O l-i 1~ ~0 CV -1 _i ~ 0 . _ _ _ _ _ _ ~ ~ o 0 _ o 0~ U~ o o Lr. o o :E m bO C~Lt~ O al tr~ o 0 ~! .
o ~Q e _ ~I _ _ ,1 ~ 0 e ~a ~ ~ ~
~ ~ ~ ,1 ~ ,1 ~ o O ~ ~ ~o ~0 ~0 ~ g ~
~ ~ ~ ~ ~ ~ ~ a~
~ m p: al a: ~ ~q ~ ~ ¢

~1 .

.

.
, s~

--78, , ____ __ . . ~. . ___ . ~î
O ~D ~ ~U~ ~)O N ~ ~
~1 O~ C~ ~D O~ In ,~
. ~O N VO O
_ _ ._ _ __ .
~ Q~ ~ tD ~
O O ~'1 ~U 00 ~0 _ S: _'- _ ____ _ _, _ ~ ~D ~1 ~ C~
~ O ~ 0 ~ ~D~ 0~ ~ ~ ~1 ~ ~D
~ ~ m O ~ r l ~ . r-1 ~ ~ N ~ O
O 1~ 1~ O ~i C~
.___ _ . ._ ~__ ' V
CU ~ 0 C~ P~ 0 ~ C~ ~
~ :~ ~ ~ o~o~ a:,l ~ r~ ~ a) ~
~ ~ O ~ ~(;~ ~t:~ ~ ~\DN~ ~r Q) ~ ~0 ~ O 0i _ ~i ~i __ ~ ~ .
m ~ ~ 'la~
_ U~ ~ ~ __ U~
O ~.D . ~ ~ N . ~ . ::~ . ~ C~
~_ _ ~ ~ C~i ~ ~D~ S: s:
__ _ _ _ _. _. ,~ O
L~. . C~ Ir~ P. l~
C~ ~~ ~CJ ~ ~ ~_ O~ ~ O
~ U~~ ~1 ~ ' ~ ~\~D h C~
In~ IU'~ I 0~ ~ ~ ~ ~ . ~ I ~ O~ O
~1~) CX:) Ir~ 'nJ If~ ~ ~ h ~rl C~ O L~ C~ ~ ~
, ,~_~ _.................. _ _ _ __ . ___ ~ al ~ ~a .
alLt~ o o Lr~ o C) h ~ ~C) 0~ bl)C\J Il~ O CU Ir'\ O
~3 ~1 r~l P~
. . ,~ . . .. .. _ . _ .0 O O ~ O O ~ ~a OC~J ~ ~ CU ~J M O
P- P~ l4 ~ P~ ~4 P~ 9, ~
. a ~ ~ ~ ~ ~ ~ ~ ~ ~

- ;.. ., . . . ~ . ~ ... . .. -. ~ .
_ _____.
. _ ~ _ o c~ ~1 ,i Lr~0' o î- o~
~ ~i . I ~ i . . . ~ <, I
r-~ V O O N ~ O CU0 ,, ~_ _~ _ _, ~ .
~, ~_ ,_ ~ ~ ~
N lt~ ~ I C~ C~ C~ I
CU . co ~ `.=~ ~ ~:U 0 ~1 . ~ ~ O ~ C'J ~ '~ ~i ~ o _ _ _ ~ _ _ __ __ .___ _ _____ , ~D C~ ,_ _~ ,ZB c~ u~ a:~ ~ ~
~'I1;~ CU I ~1 1 C I I 1~ 1 N I ~^
~ ~ ~.0 . ~ ~1 ~ ~ C~l ~D
bD ~ 0 rt N _i ~1 ~ I O
~3 ~ , _ ~_ _ ~- ~_~ _ ~CU
: ~ ,_ C~ _~ ~ ~
1 c~ ~ ._ CU CU ~ L~
a~ ~ C~ ~ ~1 ~ t- c~
:~\ :~ O ~ I ~ ~ :t I 1~ 1 It~ I 0 1 q ~ It)O ~ ' C;~ ~D ~ C~ . ~ ~D
,t~ ~1 ~ ¢ -10 C~ ~ ~i 1~,~ ~ ~
. ~ ~ _ _ _ ~ 0 ~ _~ ,_ ~ c8 ~
,., m C ~D ,~ ô~ ~ ,~ ~i , ~ ~ OC~i U~o ~1 U~O~ OOt- ~1 ~ O N 15~ 15~ _~ ~1 1~ ~ /~1 ~ 1<~ ~D ~ ~ ~
1~ ~i ~ t- CU ~ 1~ _1 ~ ~_ ~_ ~ _, _ ~ q~
.. .. - - .
-~ ~ ~- ~ ~u ~
O`\ ~D Ir~ CU . h ,_1 . ~ . .
O~ ~ O~ ~D ~ cr~ ~ ~1 o u~
. I. . . I . I . I . I ~ ~ ~Q O
~1 ~1 ~1 ~D ~ ~ a~ ~ ~ c-~
. ~i ~ ~t ~:U C~J ~ ~!R h . . . _ ~_ _ _ _ _~ h'\ Id _ . _ ___ ,~j ~ O

_~ ~
~ ~ 1~ o o Ir~ o o h ~ ~D
0 ~3 C~ 11'~ rl C~ IS~ O a3 ~
-- ~1 ~ CO 11 _._ h ~1 5~
__ _ Ir\ ~\ ~ ~O ~O ~ C ~ P' ' ~: ,1 ,1 ~1 pr~ t~ t~ S:, O
:~ O O C~ O O O :~
O CU ~U C~ CU ~U C~l O
~ ' ~ ~ ~ P~ F4 O
t~ a: a: m .

, ~S~

q~he compounds of the present is~vention are thus useful, given orally and parenterally, for enhancing the ef~ecti~reness of 5-lactam antibiotics against ~-lactamase prod~cing bacteria. On a weight basis, the dosage is from one-fifth to five times, and preferably equal to, that of the ~-lactam antibiotics. As an example, the compounds of the present invention as shown above when used in a 1:1 ratio markedly improved 'che acti~.rity of ceforanide and amoxicillin against ~-lactamase produ~ing stra~ns o anaerobic Bac~eroides SUC}l as B. fragilis, B, 'chetaiotaomicron and other species of that genus and also against resistant Staphylococcus aureus. ll~e compounds of ~he prese~nt __ invention are giverl either in admixture with or concomitantly wi~ 'che ~-lactam antibiotic with the dosage within the indicated ratio to the knowrl and customary dosage of the antibiotic .

.
Thus, the ability of the compounds of the presen~
invention to e~hance the effectiveness o~ a B-lactam anti-biotic sgai~st certain ~-lactamase-producin~ bacteria makes them valuable for co-administxation with certain ~-lactam antibiotics in the treatment of bacterial infections in ~ammals, particularly man. In the treatment of a bacterial infection, a compound o~ the present invention can be co-~ingled with the ~-lactam antibioti~, and the two agents thereby administered simultaneou~ly. Alternatively r a ccmpound of the present invention can be administered as a ~epa~ate agent during a courqe of treatment with a B-lacta~ antibiotic.

, ~

J~ ~

When USiDg a compound of the present invention or salt thereof, to enhance the antibacterial activity of a ~ -lactam antibiotic, it can be adminis tered alone, or preferably, ~ ~ormula~ion with standard phannaceutical carriers and diluents. A compound of the present invention which is in ~e acid fonn or.as a pha~naceutit:ally-acceptable salt ~ereof, can be administered oral ly or parenterally; a compound o~ the present invention in the form of an es i:er w~ich is readily hydrolyzable in v vo, is ~est administered orally. Paren~eral administration includes intramuscular, subcutaneous, intraperitoneal and intra-venous ad~inistration~, When a oompo~d of the present invention is used in the presence of a ~ar~ier or diluen~, ~aid carrier or diluent is chose~ on the ba-~is of the intended mode o admi~is~ration.. ~or example, when considerLng the or~l.
mode of administ:ration, the compound can ~e used ~ the :form o~ ~ablets; .capsules, lozenges, troches, powders, .
syrups, elixirs, aqueous sol~tions and su~pensions, and the like, .in accordance with standard phaxma~eutical practice.
_ _ .. _ . ~ .. . . . . . . _ .. .. .. . _ .. , .. . ~ . .
me proportional ratio of active ingredients to carriler will naturally d~pend on the chemical nature, solubili~y, stabi}ity and potency of ~he active ingredients, as well as the dosage contemplated. However~ these pharmaceutical compositions will likely contain from about 5% to about 80% of carrierr In the case of tablets for oral use, carriers which are commonly used includ~ lactose, sodium citrate and salts of phosphoric acid. Various disintegrants such as starchO and lubricating agents, such as magnesium stearate, ~odium lauryl sulfate and tal~, are commonly used in tablets. For oral administration in capsul~ form, useful diluents ar~ lactose and high molecular weight S~3~98 polye~hylene glyc:ols. Wherl aqueous suspensions are required for oral use, the active ingredients ~re conlbined with emulsifying and suspending agents. I~ desired, certain sweetening and/or f lavoring agents can ,be added . For parenteral administration, which includles intramuscular, intraperitoneal, subcutaneous and intravenous use, sterile solutions of the aoti~e ingredients are usually prepared, and the pH or the solutions are suitably adjusted a~d buffered. For intravenous use~ the total concentration of solutes should be co~trolled to render the preparation isotonic.

Although the prescribing physi~ian will ultimately decide ~he dosage to be used in a human sub ject~ the ratio of the daily dosages of a compound of the present invention, or salt thereof, and the ~lactam antibio~i~ will normally be in the ra~ge from about 1:5 to 5:1~ and preferably abou~
1:1. Additionally, the daily oral dosage o~ each ~omponent will normally be i~ the range from about 10 to about 200 mg~
per kilogram of body weight and the daily parenteral dosage of each componen~ will normal:Ly be about 10 to about 100 mg~, per kilogram o:E body weight. q~hese figures are illustrative only, however, a~d in some cases it may be necessary to use dosages outside these limits.

Thi~ invention is capable of indus~ial applieation .

.

Claims (9)

1. A process for the preparation of the ester compounds having the formula or wherein R1 is hydrogen or hydroxy and R2 is hydrogen, hydroxy, methyl, methoxy or chloro, wh.ich process comprises the treatment with acid of a solution of a compound having khe formula wherein R is or wherein R6 is hydrogen or hydroxy and R7 is hydrogen, hydroxy, methyl, methoxy or chloro and R3 is alkyl, aralkyl or aryl, R4 is hydrogen, alkyl, aralkyl or aryl and R is alkyl, aralkyl, aryl, alkoxy, aralkoxy, or aryloxy or wherein R8 and R9 are each hydrogen, alkyl, aralkyl or aryl or, when taken together with the nitrogen atom, are piperidino or morpholino and said treatment with acid is carried out in an organic solvent or in aqueous or partly aqueous solution at room temperature.

2. The process of claim 1 wherein R3 is methyl, R 4 is hydrogen and R 5 is methoxy, ethoxy or methyl, in the starting materials.

3. The process of claim 1 wherein the reaction is carried out in acetone or chloroform.

4. The process of claim 1 wherein the reaction is carried out at a pH between 1 and 5.

. 5. The process of claim 2 wherein the reaction is carrier out at a pH between 1 and 5.

6. The process of claim 3 wherein the reaction is carried out at a pH between 1 and 5.

7. An ester having the formula wherein R is or wherein R1 is hydrogen or hydroxy and R2 is hydrogen, hydroxy, methyl, methoxy or chloro, whenever prepared by the process of claim 1 or by an obvious chemical equivalent thereof.

8. An ester as in claim 7, whenever prepared by the process of claim 2 or by an obvious chemical equivalent thereof.

9. An ester having the formula wherein R is or wherein R1 is hydrogen or hydroxy and R2 is hydrogen, hydroxy, methyl, methoxy or chloro, whenever prepared by the process of claim 3 or 4, or by an obvious chemical equivalent thereof.