Note: Descriptions are shown in the official language in which they were submitted.
~ 753~
I. De~cription:
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 useful as an inhibitor of ~-lactamases.
The pres~med association between the resistance shown
by ~-lactam antibio~ics to certain bacteria and the ability
of those bacteria to produce ~-lactamases has led to an
in~ensive search ~or ~-lactamase inhibitors. Cla w lanic
acid is an example of such a compound presently under
intensive study. Another ~-lactamase inhibitor has in
its acid form the structure
C~OH
and is disclosed in European Patent Application 2927
published July 11, 197~.
: The compound having the structure
E~2 N
COOH
is disclosed in U.S. Patent~ 4,036,347, 4,009,159,
3,993,646, 3,989,685 and 3,954,732.
, '
-2-
1~7S8~7
U.S~ Patent 4,155,912 describes 2-penem-3 carboxylic
acid compounds having the formula
H3
COOH
and esters and salts, and see also Fa~ndoc Abstracts
82090A, 10336B and 44337B.
The co~pound (under the number CP 45899) having the
~tructure
.:
~ 5 ~ R3
: ~ . ~ 3
N
Coo~
is an irreversibly acting ~-lactamase inhibitor with
excellent solu~ion s~a~ility. It has weak antibacterial
activiky and potentiates the in v1tro and in vivo activities
o~ ampicillin versus ~-lactamase-producing strains ~A.. R.
English et al., Antimicrobial Agents and Chemotherapy, 14,
414-419 (1978), Aswapokee et al., J. Antibiotics 31~12),
1238-1244 tDec. 1978) and Derwent's Farmdoc Abstracts
89627A and 73866B~
--3--
It is disclosed by B. Baltzer et al., Mutual Pro-
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-intestinal tract and after
absorption hydrolyzed with simultaneous liberation of the
active components. As a result high blood and tissue
levels o~ 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
.
NH ~ ` ~ n-CH=N
N ~ N
O O CO O O O
F~ CH2 o~ ~co o
~7~07
--4
It is stated in GB 2044255 published October 15, 1980
that the invention therein relates to hitherto unknown com-
pounds of the general formula l:
H H
Rl- CH -OO -NH
H C - O ~ CH - A
O E~ 1
.
in which Rl stands for a phenyl~ 4-hydroxyphenyl, 1,4-
cyclohexadienyl or a 3 thienyl group; R2 repre~ents 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 of a ~-lactamase inhibitor containing a
~ -lactam ring as well as a carboxy group, A being
-~ connected via the carboxy group.
~ The new compoun~s are use~ul 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 the
acid having the formula
758~7
o o
5~ CH Cl
~1 J~ H
~ _ N
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,
potassiwm, calcium and magnesium, the ammonium salt and
substituted ammonium salts, e.g. salts of su~h nontoxic
amines as trialkylamines ~e.g. triethylamine), procaine,
dibenzylamine, N-benzyl-~-phenethylamine, l-ephenamine,
N,~'-dibenzyl-e~hylenediamine, dehydroabietylamine, -
NjN'-bis(dehydroabietyl)ethylenediamine, N-(lower)alkyl-
piperidine (e.g~ N-ethylpiperidine) and other amines
which have been used to form pharmaceutically acceptable
salts of penicillins and cephalosporins. The most
preferred salts are the alkali mekal salts, i.e. the
sodium and potassium salts, and the ammonium salt.
As used herein the term "physiologically hydrolyzed
esters" refers to those pharmaceutically acceptable esters
known in the art to hydrolyze to the free acid fo~l ln vivo.
Examples of suitable physiologically hydrolyzed esters
include phenacyl, acetoxymethyl, pivaloyloxymethyl,
a-acetoxyethyl, ~-acetoxybenzyl, ~-pivaloyloxyethyl,
S8~7
phthalidyl(3-phthalidyl)~ indanyl(5-indanyl)~ methoxymethyl,
benzoylo~ymethyl, a-ethylbutyryloxymethyl, propionyloxy-
methyl, valeryloxymethyl, isobutyryloxymethyl, 6-[(R)-2-
amino-2-phenylacetamido]-3,3-dimethyl-7-oxo ~r 4-thia-1-
azabicyclo[3.2.0]heptane-2-carbonyloxymethyl and
6-r(R)-2-amino-2-p-hydroxyphenylacetamido]-3~-dimethyl-7-
oxo-4-thia-1-azabicyclo[~.2.0]heptane 2-carbon~loxymethyl.
The preferred esters are the acetoxymethyl, pivaloyloxy-
methyl, methoxymethyl, phthalidyl~ 5-indanyl~ 6-~(R)-2-
~mino-2-phenylacetamido]~ dimethyl-7-oxo-4-thia-1-
azabicyclo[~.2.0]heptane-2-carbonyloxymethyl and
6-~(R)-2-amino-2-p-hydroxyphenylacetamido]-~3-dimethyl-7
oxo-~-thia-l-azabicyclo~.2.0]heptane-2-carbonyloxymethyl
There is further provided by the present invention
the process ~or the production as the desired product
of the acid having the formula
O O
~2Cl
N
~OOH
or a pharmaceutically acceptable salt of said acid which
compr~qes the consecutive steps o~
a) catalytically hydrogenating, as with a
precious metal catalyst such ~s palladium, an ester
having the formula
5~
--7--
CH2Cl
~ C02Rl
wherein R is benzyl or su~stituted benz~l and then
b) su~ject~ng the hydrogenated product to
oxiaat~on to produce said desired acid or salt thereof
and then, i~ desiredg
c) esteri~ying said acid or salt thereo~ to
: produce an easily hydrolyzed ester o~ said acid.
mere is furthex provided by the present
invention the process ~or the production as the desired
product o~ the acid having the ~ormula
O o
C~2Cl
ld
~OOH
or a pharmaceutically acceptable saIt o~ said acid which
comprises the consecu~ive steps of
a) oxidizing, as with KMhO4, H202 or like
peroxide or per~cid~ an est~r having the ~ormula
Br ~ C ~ Cl
~CE~ ,
~ - C--OC ~ CCl~
o
~5~7
to produce an ester sulfoxide having the formula
O O
B~ CH
'~f~S~r~C~ ~
~ N ~,
C-OCH2CC13
and then
b) reacting said ester sulfoxide with a metal
in acid~ as with zinc in glacial acetic acid, to produce
said desired acid or salt and then, i~ desired,
c~ esterifying said acid or salt thereo~ to
~ produce En ezsily hydrolyzed ester of zaid acld.
::
,
s~
A wide variety of oxidants known in the art
for the oxidation of sulfides to sulfones can be used.
However, particularly convenient reagents for alkali
met~l permanganates, e.g. potassium permanganate,
and organic peracids, e~g. ~-chloroperbenzoic acid.
.. ._ . _ .
Particularly useful pro~ecting groups for Rl are
the benzyl group and substituted ben2yl groups, ~specially
4-ni~robenzyl. Benzyl and substîtuked benzyl groups can
be removed conveniently ~y catalytic hydrogenation. In
this case, a solution in an inert solvent of the compound
of the formula A, wherein Rl is benzyl ox substituted
benzyl, is stirred or shaken under an atmosphere of
hydrogen, or hydrogen mixed with an inert diluent such as
nitrogen or argon, in the presance of a catalytic amount
of a hydrogenation catalyst~ Convenient solvents for this
hydrogenation are lower-alkanols, such as methanol; ethers,
such as tetrahydrofuran and dioxan; low molecular weight
esters~ such as ethyl acetate and butyl acetate; water;
and mixtures of these solvents. However, it is usua:L to
choose conditions under which the starting material is
soluble. The hydrogenation is usually carried out at a
t~mperature in the range from about O to about 60 C. and at
a pressure in the range from about 1 to about 100 kg,./cm.2.
The catalysts used Ln this hydrogenation reaction are the
type of agents known in the art for this kind of trans-
~ormation, and typical examples are the noble metals, such
as nicke~, palladium, platinum and rhodium~ The catalyst
i~ usually present in an amount rom abou~ 0~01 to about
2.5 weight-percent, and preferably from about ~ o about
1~0 weight-percent, based on the compound of formula A.
It is often convenient to suspend the catalyst on an inert
support; a par~icularly convenient catalyst is palladium
suspended on an inert support such as carbonO Addition-
..~
; ~75~
--10--
ally it is usual to bu~fer the reaction mixt.ure in orderto operate at ~ pH in the range ~ro~ about 4 to 9, and
preferably fro~m 6 to 8. Borate and phosphate buf~ers
are commonly used. The reaction typically takes about
one hour.
There is further provided by the present inventlon
the esters ha~ing the formula
. -~
C~3
N'X2 ~C=O
~3 0
F~Cl C~12
N "
O ~--
'3
O
wherein R is ~ ' a ~
2 S
or Rl ~ wherein Rl i9 hydrogen or hydroxy and
R~ is hydrogen, hydroxy, methyl, methox~ or chloro and
~s~a~
preferably the ester havin~ the ~ormula
~\ S C~
Q~ CH-CO-NH ~
NH2 ~$
O 'C=O
O O
- ~ "" C~2Cl
~ C:H3 7H2
O ~ O
O
. and
the ester having the ~ormula
CH3
~0~ Il~_CO_~
OJI O
~ ~ Cl '~2
O ~ -O
8 and the
process ~or producing such an ester which comprises the
treatment with acid o~ a solution of a compound having
the ~ormula
.;` ` ~3L7~7
--12--
/ \ ~C=O
R3
OA
F~ CH2
_o
O
where~n R is
R5
or R ~ wherein R4 is hydrogen or hydroxy and
R5 is hydrogen, hydroxy, methyl, methoxy or chloro and
R1 i~ alkyl, aralkyl or aryl~ R2 is hydrogen, alkyl, aralkyl
or aryl and R3 is alkyl, aralkyl~ aryl, alkoxy, aralko~y,
aryloxy or R4
-N ~ 5
wherein R4 and R5 are each hydrogen, alkyl, aralkyl or aryl
or, when taken together with the nitrogen atom, are piperidino
or morpholino with said treatment with acid preferably being
carried out in an organic solvent such as acetone or
chloroform or in aqueous or partly aqueous solution and
pre~erably between pH 1 and pH 5 at room temperature.
. ~ -13-
`` il7580i~
It ~s further preferred that, in the amino-
protecting group, Rl is methyl, R2 is hydrogen and R~
is methoxy~ ethoxy or methyl; this requires the use of
methyl acetoacetate, ethyl acetoacetate or acetylacetone.
In the remo~al of the a-amino-protecting group
it is preferred that use be made of a strong mineral
acid such as hydrochloric acid or of formic acid.
: mere is further provided by t~e present invention
as a novel intermediate an ester having the formula
. . .
Br~CH zCl
~ OR
wherein R is benzyl or substituted benzyl, and preferably
~p~nitrobenzyl, and the process for lts production which
- comprises heating, pre~erably at reflux, a compound
having the formula
~r
~ ~S~ .
O ~C-OR
O
wherein R is benzyl or substituted benzyl and pre~erably
p-nitrobenzyl, in an ~nert, anhydrous organic solvent,
preferably dioxane, in the presence o~ large, and preferably
` -14-
S8~7
equimolar, amounts of a weak tertiary amine~ preferably
quinollne, and an acid chloride, preferably benzoyl
chloride, until the reaction is substantially complete.
There is provided by the present lnvention in
addltion as a novel intermediate an ester having the
formula
O
lir~_~2Cl
~-OR
wherein R is benzyl or substituted benzyl, and preferably
p-nitrobenzyl, ~nd the process for its production which
comprises oxidizing a solution in an inert solvent,
pre~erably methylene chlor~de, o~ a compound having the
formula
"~,
1~ .
O 'C-OR
o
wherein R ls benz~l or substituted benzyl, and preferably
p-n~trobenzyl, at about room temperature by the use of a
peracid, pre~erably m-chloroperoxybenzoic acid.
~:~7~ 07
-15- ` ~
mere is Purther prov~ded by the present
invention as a novel intermediate the ester having
the ~ormula
O O
Br ~ CH2Cl
'f~ S~ CH3 -'
O C-OCH2CC13
O
the process for lts production which comprises oxidizing
a solution in an inert solvent, preferably methylene
chlorid~, of a compound having the formula
C Cl
~; ~ ~ C-OC ~ CCl~
O
at about room temperature by the use of an oxidizing
Qgent such as KMnO4, H202 or like peroxide or, preferably
a peracidg preferably m-chloroperoxybenzoic acid.
: "Skellgsolve Bt' iB a petroleum ether fraction
o~' b.p. 60-68C. consisting essentiall~ of n-hexane
("Skellysolve" i8 a trade name of the Skell~ Oil Co.).
The following examples are provided solely for
the purpose o~ illustrat~ng the preparation of representa-
tive compounds o~ the present invention and are not ko be
constr~ed as l~miting the lnvention.
f~ *Trademark
--16--
gL~lL758~'7
Example
PreParation of Potassium 2~-Chloromethyl~g3~b~
:
carboxyla~e Sulfone ~BL-P2013)
i .
O O
- 3' 1`
B~ B~
: ~ 2~ 2--C~2~NO2
: 1 2
~: :
: O
B~H2Cl Br, r_~S~CH2Cl
O CO --CH ~ O~ N ~'~CO C:H
~2
\ /
r ~ ~ 0~ ~0
r ~CH ~ rl~ ~f'
,~N - ,~, ,~ N - ~"
a C2H _ o co2x
~BL--P2 013 )
~ .
'
.
~17-
~75~CI17
&~-Bromo~e~lc 1 Acid S-Sulfoxide ~)
1. Dissol~e 30 g (37.5 mmole) ~-bromopeniclllanic
acid N~N'-dibenzylethylenediamine salt [G. Cignarella et al.,
J. Org. Chem. 27, 2668 (1962) and E. Evrard, Nature 201,
1124 (1964)~ in 3~0 ml o~ methylene chloride. Agi.tate and
cool to 0C.
2. Slowly add 1~ ml (156 mmole) concentrated
hydrochloric acid into the methylene chloride solution.
me precipitation of dibenzylethylenediamine HCl salt
~DBED~HC1) takes place within a minute. St~r the slurry
at 0-5C for 10 minutes.
3. Filter to r~move 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 ~uickly as possible.
Avoid holding the acidic methylene chloride solution for
a prolonged perlod. There may be some ~iltration problems
because of the fine nature of the p.recipitate. Addition
o~ ~ilter aid to the slurry may be help~ul.
4. Wash the combined methylene chloride filtrate
~ld wash with 60 ml o~ cold water~ Agitate 5 minutes ~nd
dlscard the aqueous phase. The pH o~ the wash is 2.0-2.3.
5. The methylene chloride æolution containing
6~-bromopenicillanic acid is concentrated under reduced
pressure to a ~olume o~ 65-80 ml. Cool ~nd stir the
solution to 5C.
*Trademark
~æ
~ -18-
~5 51~
6. With vlgorous agitation cautiously add 13 ml
(86.9 mmoles) of 40~ peracetic acid over a period of 30
minutes.- The reaction is exothermic. ~aintain temperature
between 15-18C with ice bath cooling. The sulfoxide begins
to crystallize after 10 ml peracetic acid is added. Cool
and stir the slurry at 0-5C for two hours.
7. Fîlter and wash the snow white cake with the
following sequence: 10 ml 5C water, then 10 ml 0-5C
methylene chloride, and finally wash with 15 ml of heptane.
8. Dry the cake in 45C air oven to const~t
weight3 about 6-10 hours should be sufficient. Extended
heating may generate a trace of pinkish color. The weight
of 1 is about 16.26 gm, 73.24% yield.
9. The reaction mix and final product may be
monitored by TIC using 15 toluene/~ acetone/l ~cetic
acid ~HAC) or 8 acetone/8 methanol/3 toluenejl HAC solvent
systems, The final product should be analyzed by NMR and
IR.
p-Nitrobenzyl 6a-bromopenicill~n~' e S 9~1r.~ide (2)
To a solution of 12 g (0.04 mole) of 6a-
bromopenicillanic acid s-sul~oxide in 100 ml of acetone
wa~ added 7.5 g (0.041 mole) of potassium 2-ethylhexanoate.
The salt was collected by filtration~ washed with cold
acetone and air dried to yield a total of 10 grams. The
crystalline potassium salt was dissolved in 75 ml of
dimethylacetamide and 7.8 g (0.04 mole) of p-nitrobenzyl
bromide was added. m e solution was stirred at 23C for
24 hours. The mixture was diluted with 500 ml of water
and extracted ~ith ethyl acetate. The ethyl acetate layer
--19--
~l 3L751~3~7
was washed four times with water and dried over anhyàrous
magnesi~am sulfate. The solvent was evaporated at 35C
(15 mm) to an oil which crystallized. The light tan
crystals of 2 were slurried with ether and collected by
~iltration to yield 9 g (70%) mp 124-125C dec.
Anal. calc'd for C15H15BrN206S: C, 41.98; H, 3.05; Ng 6.52
Found: C, 42.00; H, ~5.48; N, 6.98
IR(KBr): 18QO(s), 1740(s), 1610(w) 3 1520(s), 145~)(mj,
1350(s), 1060(m~, 740(m) cm~l. H-N~ (60 ~z, DMS0): ~1.22
(s,3H), 1.6 (s,3H), 4.67 (s,lH), 5.2 (d,J~1-5 Hz,lH),
5.45 (s92H), 5.68 (d,J~1-5 Hz91H), 7.5-8.5 (m"4H).
p-Nitrobenz~l 2B-chloromethyl-2a-methyl-6-bromopen~-;~a-
carboxylate (3)
A solutioll o~ 5 g (0.012 mole) of p-nitrobenzyl ~
6a-bromopenicillanate S-sul~oxide (2) in 120 ml anhydrous
dioxane was heated at reflux under nitrogen for 4 hours
with 1.5 g (0.012 mole) o~ quinoline and 1.6 g (O.Q12 mole)
of benzoyl chloride. me solution was diluted with 600 ml
of water and extracted with ethyl acetate. me ethyl
acetate extract was washed with 5% sodium bicarbonate
solution, 5% phosphoric acid solution and finally with
water. The organic layer was dried over anhydrous
magnesium sul~ate and evaporated to an oil at 35C (lS mm).
~he oil crystallized and was collected, washed with ether,
and ~nally with cold toluene to yield 3, 3.5 g (65~) mp
130-135C dec~
r C15H15ClBrN205S: C, 40 . o6; H, 3.14; N 6 23
Found: C, 40.19, H, 3.12; N, 6.75
IR~Br): 1792ts), 1740(s), 1610(w), 1520(s), 1~53(s),
1280(m), 1025(w), 990(w), 750(w) cm 1.
N~ ~60 mHz, DMS0): ~1.45 (s,3H), 3.5-4.~3 (m,2H)~
5.05 (s,lH), 5.4Z (s,2H), 5.5(d,J~1.5 HZ~lH)~ 5.62
(d~J~1.5 Hz,lE), ~.5-8.5 (~,4~I).
-20-
S~l~7
p-Nitrobenzyl 2~-Chloromethyl-2a-meth~lpenam-6a-carboxylate
Sulfoxide (~)
_
A solution of 1 g (0.0022 mole) of p-nitrobenzyl
3~-chloro~ethyl-2a-methyl-6a-bromopenam-3a-carboxylate
(3) dissolved in 50 ml of methylene chloride was stirred
with 473 mg of (0.0022 mole) of m-chloroperoxybenzoic acid.
me solution was stirred at 23C for ~ hours. The methylene
chloride was e~aporated to 0 ml at 15 mm and ~3C and the
concentr~ted solution was diluted with 50 ml of heptane
("Skellysolve B") The solvent was decanted and the residue
was slurried with ether and (4) soon crystallized yield
250 mg, 24% mp 136-1~7C dec.
Anal- Calc'a for C15H14BrClN26S G~ ~8-68; H~ ~-02; Ng 6-02
Found: C, ~9.14; ~, 3.13~ N, 5.96
-
IR(KBr): 1800(s), 1760~s), 1520(s), 1~50(s), 12QO(s),
1050(m), 830(w)~ 740(w) cm 1~ H-NMR (60 mHz, DMS0):
~ 1-32 (s,3H), 3.8-4.5 (m~2H), 4.97 (s,lH), 5-25
(d,J~1.5 Hz,l~ 5.45 (s,2H), 5.6 (d,J~1.5 Hz,lH),
7.8-8.5 (m,4H).
Potass u ~ -Chlorometh~1-2a-methylpen~m-~a carbox~late
Sulfone (5) (BL-P2013)
To a solution of 7 g (0.015 mole) of p-nitrobenzyl
2~-chloromethyl-2a-methyl-6a-brvmopenam-3a-carboxylate
sulfoxide (4) in 150 ml of ethyl acetate wa~ added a
suspension of 4 g of 30% palladium on diatomaceous earth
("Celite") and 2.8 g o~ sodium bicarbonate in 150 ml of
water. The mixture was hydrogenated for ~ hours at 50 psi.
-21
~5~7
me catalyst was separated by filtration and the aqueous
layer was separated ~nd treated with 1.5 g o~ potassium
permanganate in 50 ml of water. me mixture was stirred
for 1 hour and 250 mg of sodium bisul~ite was added. The
mixture was ~ltered and the filtrate wa~ adjusted to
pE 2 with concentrated hydrochloric acid. The solution
was lyophilized to give a white amorphous powder. me
solid was extracted with ethyl acetate, evaporated to
volume of 20 ml and diluted with 100 ml of heptanes
("Skellysolve B"). White, hygroscop~c, solid 2~-
chloromethyl-2a-methylpen~m-3~-carboxylic acid sulfone
was coll~cted. The acid was dissolved in acetone and
treated with solid potassium 2-ethylhe~anoate. A
crystalline white salt precipitated to give, after
filtration, 170 mg o~ 5 mp ~140C dec.
C8~7ClKN05S 2H20: C, 28.27; H, 3-24 N 4 12
Found~ C, 28.27; H~ ~.69; N, ~.84
IR(KBr): 1790~s), 1770~m), 1620(s), 1460(m), 1~70~s),
l~lO(s), 1200(s), 1140(s), 955(m~, 740(m) cm 1 H-NMR
(100 mHz, D203: ~1.68(s,~H), 3.2-~.9 (m,J~2 ~z7 J~4 Hz,
J~6~Hz~H)~ 4.0-4.4 (m,2H), 4.3 (s,lH), 5.02 (d d, J~ Hz),
J~2 Hz,lH).
Example 2
Pivaloyloxymethy~l 2~-Chloromethyl-2 ~ -3a-
carboxylate Sulfone
2~-Chloromethyl-2a-methylpenam-3-carboxylic
acid sulfone in dimethylformamide is treated with one
equi~alent of triethylamine and stirred to ef~ect solution~
~romomethyl pivalate (1 equivalent) in dimethylformamide
--22-- -
~L7SI3~7
is then added~ The resulting mixture is stirred at room
~mperature. m e mix~ure is then clarified by filtration
and ~he filtrate poured into ice w~ter. The separated
solid is collected by filtration, washed with water and
dried to give the title ester.
me respective acetoxymethyl t methoxymethyl,
acetonyl and phenacyl esters of the same acid are
prepared by substituting in the method above for the
bromomethyl pivalate used therein an equimolar weight of
chloromethyl acetate, chloromethyl methyl ether,
ehloroaceton~ and phenacyl bromide, respectively.
Example 3
Piva oy_oxy~ethyl 2~ Chloromethyl-2a-methylPenam-3~-
carbo~ te Sulfons BL-P2024
~ H3
~S~ ~, O
~ I~CH2C1 ~ NaI + ~0 ~ ClCH2-0-C-C(CH3)3 ace~one
O ~306) C2}~ '~
~BL--P2013 ~
5~ 7
~lzCl
0 CO2CE12-O-C-c(c~3)3
5BL-P2024)
. - ....
.~
; ~ To a stirred suspension of 14.6g (0.0487 mol)
of BL-P2013 (5) in 200 ml of acetone was added 4 ml of a
10% aqueous solution O.æ sodium iodidP and the mixture was
: :brough~ to reflux on the steam ~akh~ To ~his refluxing
suspe~ion was added 14~8 ml (0.1 mol~ of redistilled
chloromethyl pivalate ~B.P. 34C at 7 mm ~g) all at once.
~he mixture was stirred at reflux for three hours and then
coole~ to room temperature ~22C). The crystalline solids
: were collected by filtration, washed with 3 x 30 ml of
: acetone and the combined filtrates were evapora~ed in an
oil in vacuo at c22C. The oil was then taken up in 500 ml
of ethyl acetate and washed once with water (200 ml) and
once with saturated Na~S04 while being stirred with 2 g.
o decolorizing carbon with cooling tice bath). After 20
minute$ ~he mixture was filtered through a di~tomaceous
(Dicalite) pad with suction and the pad was washed with
4 x 100 ml of e~hyl acetate. The combined ~iltrates were
concen~rated in vacuo at 22~C to an oil. The oil was then
, --24--
7S~7
concentrated further a~ about 22 C and c 1 mm Hg to remove
most of the residua~ chloromethyl pi~alate. The remaining
oil was then triturated twice with 50 ml portion~; of
n-pentar~e and then lef t over the weekend in the cold room
~about 10C) under n-pentane. The solid crystalline mass
was then broken up to a solid powder under 40 ml of
4:1 mixture o ether-n-pentane. The product was then
collected by filtratio~, washed with ether-pentane ~
~hen pentane and air dried. Af~er drying in vacuo for
four hours over P205 there was obtained 13.37~. of
pivaloyloxymethyl 2~-chloromethyl-2a-methylpenam-3a-
carboxylate sulfone (about 75~ yld) M.P. 93~-~5~C.
~nal- Calcd- for C14H20ClN~7S C, 44.03; H~ 5.27; ~ 3~7-
Found: C, 44.11; H, 5.08; N, 3.85.
Exam~le 4
Recrystallization of Pota~sium 2~-Chloromethyl-2~-
methvloenam~3~-carboxvlate Sulfone (BL-P2013)
To a mixture of 20 ml o~ n-butanol and one gram
of BL-P2013 (5) was added water, one ml at a tLme, with
shaking in a separator~ funnel until a pale yellow solution
was obtaîned. The clear solution was ~ ered through
a fluted ~ilter paper and the flask and fil~er paper washed
with about 10 ml of 9:1 n-butanol-H20 and the combined
filtrates were diluted with a further ~0 ml of n-butanol.
The resulting ~olution was placed in a round-bu~omed
f lask on the roto-vap and evaporated under reduced pressure
to approxi~ately one hal~ the original volume. The snow
white crystalline product was collected by filtration, washed
.~
-25 -
~S~7
with 6 x 10 ml of acetone and air dried. Yield 810 mg.
After vacuum drying 6 hours over P205 at <1 mm Hg.
there was obtained 800 mg M.P. 215C ~dec.) (80% yld).
Anal. Calcd. for C8HgClN~5SR-lHzO: C, 2g.67; H, 3.39; N, 4.63;
Cl, 10.94; K~F.~2~ 5.56.
Found: C, 29.23; H, 3.38; N, 4.49;
Cl, 10.74; ~oF~H20~ 5.74.
This recry~tallization procedure produces a
crystalline monohydrate differing from the ~tarting
material which is essentially anhydrous.
_.
Example 5
o, 11
C ~ 0-C-Cl ~ ClCH2-0-C-Cl
light
¦ fClS03H
ClC ~ -0-S-Cl
6
See Chemical A~stracts 27f 2427 and 22, ~828; and
GB 299064,
.
--26--
gL~.'75~
~lS0~5CX2~1 (6 )
o o o o
~S/ C:H;~Cl ~5~ CH2Cl
1FN~ FN~g
~OOK I -OCH2
. 7
NaI
ac etone
O ~ ~0
~1
,C,-OCH2I
: ~ ~
8 ~ CO-NI~
'COOK
H C N
}[11~ ~o
OCH~
9 (U.S. Pa~ent 7,316,247)
I
--27--
~L7~ 7
~c f-- H
~ C~c~
OC~13
:', O
1 .
. 1H
::
,=~ CH~
~CH--CO--NH ~;~ ~CX3
.NH2 o C=O
"S"' CH~Cl I
0~ ' ~C~O
O
-28-
~7~8~
a) A solution o~ chloromethyl chlorosulfate (0.115 mol)
in 40 ml. dichloromethane is added dropwise, keeping
the reaction temperature below ~0C., to a solu~ion of
compound 5 (0.1 mol) and potassium bicarbonate (0,3 mol)
and tetrabutyl~mmonium hydrogen sulfate (0.01 mol) in
200 ml. dichloromethane-water (1.1). At the end
of the addition the mixture is stirred at room tempPrature
for ~0 minutes, the organic ph~se is separated ~nd the
aqueous phase is extracted with dichloromethane (50 ml.).
Th~ combined organic phases are dried (Na2S04) and evaporated
in vacuo to giYe a resldue which is dissolved in ether
(150 ml.). Insoluble material is ~iltered of~ after
adding diatomaceous earth and the ~lltrate is e~aporated
in vacuo to proYide compound 7.
b) To a suspension of compound 5 (1.5 g.) ~n
dimethyl~ormamide (12 ml.) there is added 1.6 g. bis-
chloromethyl sulfate and the mixture is stirred at room
te~perature ~or ~5 ~inutes. A~ter dilution with ethyl
acetate (50 ml.) the mlxture is washed with water and
then aqu~ous sodium bicarbonate3 dried and evaporated in vacuo
to leave compound 7 as an oil.
c~ To a solution of compound 5 (0.005 mol) in
dimethyl~ormamide (7.5 ml.) there is added triethylamine
(0.007 mol) ~nd chlorolodomethane (0.030 mol) and the
mixture is stirred at room temperature ~or four hours.
A~ter dilution with ethyl acetate (30 ml.) the mixture
is washed wlth water (3 x 10 ml.) followed by saturated
aqueous sodium chloride (5 ml.), dried and evaporated in
vacuo to leave compound 7 as an oil.
-29-
~75~3~7
d) To a mixture of compo~nd 5 (0.15 mol) silver
nitrate (0.15 mol) and silver oxide (7.5 g~) in acetonitrate
(750 ml.) there is added chloroiodomethane (1.5 mol).
After stirring for 48 hours at room temperature, the silver
salts are filtered off and the filtrate is evaporated
to dr~ness in vacuo. The residue is dissolved in ethyl
-
acetate (200 ml.) and the solution is washed with saturated
aqueous sodium chloride, filtered, dried and evapor~ted
in vacuo to give compound 7.
Compound 7 and other intermediates and final
products of the present invention are purified, i~ desired,
by column chromatography, as on "Sephadex" LH20 using
chloroform-hexane, 65:35 as the eluent ~or example or by
silica gel chromatography~ e.g. using Mallinckrodt
CC-7 and hexane-eth~l acetate~ ~:2 or ethyl acetate-
petroleum ether, 8:2 or ~:3 or 1:9 or 15:85 or ethyl
acetate-n-hexane, ~:6 or 3:1 hexane-ethyl acetate, ~:1
or 1:1 or 1:1~ or cyclohexane-ethyl acetate, 1:1.
Thin layer chromatography is also useful.
"Sephadex" is cross-linked dextran 2~(diethylamino) ethyl
2-~C2-(diethylamino)ethyl}diethylammonio]ethyl ether chloride
hydrochloride epichlorhydrin (See Merck Index, Ninth Edition,
item number 7337).
A ~olution of compound 7 (0.2 mol) and socLium
iodide (0.~ mol) in acetone (150 ml.) i5 stirred at room
temperature ~or 18 hours. The resulting suspension is
cooled to about 0C. and ad~usted to about pH 7.2 by the
addition of saturated aqueous sodium bicarbonate with
stirring. A~ter decolorizing by titratio~ with 0.5 M
aqueous sodium thiosulfate, watPr (150 ml.) is added
dropwise to the stirred mixture to precipitate solid
compound 8 w~ich is collected by filtration, washed with
acetone-water 1:1 (2 x 20 ml.), i~opropanol (2 x 20 ml.)
and ether (2 x 20 ml.) and dried
-30-
S~
~ mpicillin is converted to compo~d 9 by the
use of methyl acetoacetate in the procedures o~ U.S.
Patent 3,316,247. Then to a stirred solutlon of co~pound
(0.57 mol) in d~methylform~mide (1 li.ter) there is added
at 5C. 0.5 mol of compound 8. A~ter ~tirring for 15
minutes at 5C. the reaction mixture is poured into an
ice~cold mixture o~ ethyl acetate (4 liters) and saturated
aqueous calcium chloride (2 liters) with stirring. The
organic layer is separated, washed with saturated aqueous
c~lcium chloride (2 x 500 ml. ), filtered and evaporated
to about one liter in vacuo to provide a concentrated
solution of compound 10. To this concentrate there is
added water (500 ml. ) and n-butanol (50~ ml.) and then,
dropwise, 4 N h~drochloric acid with stirring until
the amino-protecting group is removed to pro~ide a solution
of compound 11. A~ter the addition of the acid is finished
ether (1 llter) and water (500 ml. ) are added to the ~tirred
mixture, the aqueous phase is separated and the organic
phase is extracted with water (800 ml.). me co~bined
aqueous extracts are washed with ether (1 liter) and then
sodium chloride (640 g.) and dichlorometh~ne ~2 liters) are
added and the mixture is stirred for 15 minutes. The
organic phase is separated and the aqueous phase is
extracted with dichloromethane (1 liter) and the cornbined
organic extracts are dried(MgS0~) &nd evaporated to
about 600 ml. under reduced pressure to give a concentrated
solution o~ compound 11. Addition to the concentrate
o~ 200 ml.2-butanon~ followed by cooling precipitates
solid 6-~(Rj-2-Amin~-2-phenylacet~midol~~,3-dimethyl-7~
oxo-4-thia-1-azabicyclo~3.2.0~heptane-2-carbonyloxymethyl
2~-chloromethyl-2-a-methylpenam-~a-carboxylate sul~one
(11~ which is collected by filtration.
.
~ 31
~ ~.75~7
Ex~mple 6
6-[(R)-2-Amino-2-p_hydroxyphenylacetamido]-3,3-
dimethyl-7-oxo-4-thia-1-aæabicyclo~.2.0]heptane-2-
; carbGnyloxymethyl 2~-chloromethyl-2-a-methylpen~m-~a-
~ carboxylate sulfone having the formula
,.~
rl7
HO~ CH--CO ~ C=O
O O
O
..
is produced by substituting ~moxicillin ~or the
ampicillin used in the procedure o~ Example 5.
-32-
~ ~7 ~ ~ 7
Ex~mple 7
C
H~
a) ~
~ _ ~ g .~ N-Bromosuccinimide
o
~-azo-isobutyronitrile
CC14
C~r
~ ~O
\=/ ~O
As taught by U.S. Patent 3,860,579, recrystallized
phthalide ~50 ~., 0.375 mol)-and recryætallized N-bromo-
succinimide (0.375 mol) were refluxed by 4.5 hours ln
the presence of about 100 mgm a-azobutyronitrile in one
liter GC14. m e mixture was cooled to about 15C. and
~iltered to remove succinimide which was itsel~ washed
with about 100 ml. CC14 and ~iltered. The combined CC14
phases were concentrated in vacuo to about 150 ml. giving
solid 3-bromophthalide which was collected by filtration,
washed with about 50 ml. CC14 and air-dried to yield 54 g.
which weighed 50 g, after recrystallization ~rom boiling
cyclohexane, m.p. 84-86C.
b)
. ~ ....... . . .. ._ . ,
O O
C B r -
C ~Cl ~ ~0
C-Og
0
_ DMF
22C.
~
-33-
~L~7~il3al7
o o
N ~ *
a ~l-Q-CH ~ BL-P2036
\C ~ *dl
O
To a stirred partial solution and partial suspension
of compound 5 (BL-P2013; 2.3 g., 0.0075 mol) in 20 ml.
dimethylformamide (DMF; dried at least 3 weeks over ~A
molecular seives) was &dded 1.7 g. (o.oo8 mol) o~
~-bromophthallde (12) and the mixture was stirred 4 hours
at 22C. The resultlng mixture was poured into a mi~ture
of 200 ml. ice-cold water and 200 ml. ice-cold ethyl
acetate (rinsing the ~lask with a little ethyl acetate)
and the mlxture was shaken. m en the organic solvent phase
was separated and washed with seven portions of ice-cold
water (lOO ml.). The eth~I aceta~e phaae was w~hed o~ce with
saturated aqueous Na~S04, dried in the cold over Na2S0~,
filtered and evaporated to dryness in vacuo to leave as the
residue an oil whlch was triturated twice with methylcyclo-
hexane (25 ml.), twice with "Skellysolve B'~ (b.p. 60 68C.,
essentially n-he~ane) (25 ml.) and four times with 25 ml.
n-hexane to 2.5 g. compound 13 as a nearly wh~te solid
after drying in air. This product was then dried over
P205 at less than l mm Hg to give 2.5 g. compound 13
m.p. 104C. (dec.). Its estimated purity was 85-95
-34-
~7S8~7
Anal. Calcd. ~or C16H14ClN07S: C, 51.61; H, 3.79; N, 3.77;
Cl, ~.5~.
Found: C, 52.59; X, 4.67; N, 3.21,
Cl, 7.73; K.F.H20, 0.27.
Example 8
Pivaloylox~methyl 2~-Chloromethyl-2a-meth~lpenam-3a-
carbox~Iate Sulfone
h mixture o~ 1 g. (0.0031 mole) of potassium
2~-chlorometh~1-2a-methylpenam-~a-carboxylate sulfone
hydrate and 1 g. of 3A molecular sieves was stirred in
15 ml. of dimethyla~etamide ~or 2 hours at 23~. To
this mixture ~as added ~70 mg. (0.00~1 mole) of
pivaloyloxymethyl chloride and the stirring was continued
for 18 hours. m e molecular sieves were collected and
the ~lltrate was diluted with 100 ml. of water and extracted
with ethyl acetate. The ethyl acetate was wa~hed ~ine
times with water and dried over anhydrous magnesi~m sul~a~e.
m e solvent was removed at 30 (15 mm) to leave an oil which
was chro~atographed on silica using silicar CC-7 (methylene
chloride 8, eth~l acetate 2) showing 1 spot R~ .5. The
residue obta~ned crystallized from heptane t"Skellysolve
B") to yield 100 ~e. (M.P. 94-95) of pivaloyloxymethyl
2~chloro~ethyl-2a-methylpenam-3a-carboxylate sulfone.
Anal. Calcd,: C, 44.03; H, 5.27; N, ~.67.
Found : C, 44.20, H, 5.24; N, ~.6~.
The NMR and IR spectra were consistent ~or structure.
'
~7S~7
Example 9
Sodium 2~-Chloromethyl-?a-meth~lpenam-3a-carboxylate
Sulfone
C1 , H+ NaEH~ CH~Cl
C ~ ~ (1) ~ ~ CH3
~ N ~ Na
: ~ ~o E 1 2
2 C8HgClN05SNa
(356) .(289.67
__ _ _ .
To a stirred solution of 500 mg. of B1-P201
(potassium salt) in ~ ml. of H20 and 10 mlu o~ ethyl
acetate was added 2N HCl until pH 1 was obtained (done
in an ice-bath with vigorous stirring ) . The mixture was
then saturated with Na2S04, the aqueous layer separated
and the organic phase dried brie~ly in ice over Na2S04,
filtered and treated dropwise with 50% NaEH (sodium
2-e~hylhexanoate) - in anhydrous n-butanol until neutral to
moist pH paper. Product did not crystallize upon scratching and
was then concentrated in vacuo to an oil wh$ch was dissolved
in acetone (5 ml.), scratched - no crystals, ether added to
cloud point - no cyrstals. Concentrated in vacuo on
rotovap. to an oll which was dissolved in ethyl acetate -
added one drop H20 - ~cratched no cry~tals, Concentrated
in vacuo and then residue was triturated with 5 ml. of
n-butanol - 200 mg. o~ amorphous white powder obtained, ether
washed - air dried - vacuum dried over P205 ~or 24 hours.
180 mg. final yield o~ sodium 2~-chloromethyl-2a-methylpenam-
3a-carboxylate sul~one; dec. pt. ~100 indef.
~nal. Calcd. ~or C8HgClN05SNa: C, 33.10, H, 3.13; N, 4.89
Found: C~ 33.20; H, 3.69; N, 4.44
K.F.H20, 4.04
-36-
~L7~
Example lo
Potassium ?~-chloromethyl-2-methylpenam-3 carbox~late
Sulfone (BL-P2013)
` To 10 L oi wa~er3 1~0 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-methylpen~m-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 v~r~ slow and 200 g of 10% Pd on BaS04
were added and the mixture hydrogenated until no further
signi~icant hydrogen absorption was perceptible.
The slurry was ~iltered through a diatomaceous
earth ("Celite"~ pad, the pad was washed with water and the
aqueous ?hase washed with 3 L o~ ethyl aceta~e. To the
aqueous solution, ~ L of ethyl a~etate were added and the
pH o~ the mixture adjusted to 1.5 with 150 ml o~ ~2 N HCl
at 10C. m e organic phase was separated and the aqueous
solution saturated with Na2S04 10 ~2 and e~tracted with
2 x 1 L of ethyl acetate. The combined extracts were
dried wlth magnesium sulfate. The drying agent was removed
and 260 ml of 2 N potassium 2-ethylhexanoic acid in butanol
were added at 0C.
~ ter stirring 2 hours at 0C, the potassium 2~-
chloromethyl-2-methylpenam-3-carboXylate ~ul~one (BL-P2013)
was collected and dried in n vacuum at room temperature~
Yield : 1~4.8 g (about 70%).
*Trade~ark
.
_3 j
~L~75il~7
Example 11
p-Nitrobenzyl_6a-Bromopen~ cillinate Sulfoxide
. . . , _ . . _ . . .
o
i 1~ ~ (101) ~N02
0 ~ ~C02H (216 )
(296 )
o
Br"~ ~ S
i I I
C02CH2-~ N2
~ (4~1 . 28)
... .. .
Procedure:
To 200 ml. of N,N-dimethylacetamide was added
44 g, (0.148 mole) o~ 6~-bromopenicillanic acid sulfoxide
~ollowed by 20.5 ml. (0.148 mole) of triethylamlne and
38.2 g. (0.177 mole) of p-~itrobenz~l bromide, It was
stirred at 22 for 20 hour~.
The reaction mixture was poured into 1 liter H20
and extracted into 3 x ~00 ml. o~ methylene chloride. The
combined meth~lene chloride extracts were washed with 200 ml.
o~ 5% aqueous sodium bicarbonate solution and dried over
sodium sulfate at 5 for hal~ an hour. The solution was
filtered and evaporated under vacuum to a residue. The
.
-38-
~.i7S~7
residue was diluted with ether and the solid coliécted by
filtration to yield 54 g, p-nitrobenzyl 6a-bromopen~cillinate
sul~oxide after drying.
85% y~eld.
nmr consistent for structure.
me y~eld for this step was the same as for the K-salt
esterification. The advantage is there-was no need to
make the K-salt. ~A s-tep which goes ln 85% to 90~ yield).
Eæample 12
Preparation of ~-~itrobenz~l 6a-Bromopenicillanate Sulfoxide
To 4.~75 L of N,N-dimethylacetamide was added 873.0
g (2.95 moles) o~ 6a-bromopenicillanic acid (S) sulfoxide and
then with stirring and while keeping the internal tempexature
below 35C~ 2g~ g (2.95 moles) o~ triethylamine followed by
764 g (~.54 moles) of p-nitrobenzyl bromide. The mixture
was stirred then at room temperature for 5 hours and let
stand o~ernight.
The reaction mixture wa~ poured i~to 20 L of water
and extracted with ~ x 7 L of methylene chloride. I~e
comblned organic extracts were washed 5 x 7 L of water and
then wlth 7 L of 5~ a~ueous sodium bicarbonate solution and
dried over anhydrous magnesium sulfate.
me magnesium sul~ate was filtered of~ and the
solution evaporated to a crystalline residue; 4 L of diethyl
ether were added and the crystals collected to y~eld after
drying at room temperature 1171 g (92%) of p-nitrobenzyl
6a-bromopenicillanate sulfoxide.
Br 18.48~ ~calculated 18.5~%), ~ (0.25% MeOH) I 162.
. .
-3g-
7S~
Example 13
Preparation of P-Nitrobenz~l 6a-Bromo-2~-Chloromethyl-
2-methylpenam-3-carboxylate Sul~one
To 16 L oY acetic acid was added 364.6 g (0.812
mole) of p-nitrobenzyl 6a-bromo-2~-chloromethyl-2-meth~lpenam-
~-carboxylate. To the solution so obtained and stirred at
room temperature, a solution of 282 g (1.78 mole) of KMnO4
in 26 L of water was added dropwise over 3 hours. me
mixture was then stirred at room temperature for 1 hour
and ~ 2 (37%) was added dropwise until a colorless solution
was obtained. 30 L of water were then added~ the mixture
stirred for 1 hour at ro~m ~em~erature~and th~ crystalline
precipitate was collected, washed with 3 x 5 L of water and
with 2 x 2 L of ethanol and dried over vacuum at room temper-
ature.
Yield: 297 g (76%)
(0.5% C ~C12) ~ 75.9
Example 1~
Preparation of BL-P2013 Free Acid
C~ Cl ~ pO~ ~ ~ C Cl
~C2~K C02H
:
:
-40-
~S~
To a mixture of 25 ml o~ ethyl aceta~e and 10
ml of water was added 800 mg (0000261 mole~ of B~-P201
potassium salt. After all of the solld h~d dissolved5
the mlxture was treated dropwise with 50~ aqueou~ phosphoric
acid w~th vigorous shaking until no more material precipitated
from the aqueous layer. The ethyl acetate layer was
separated, then washed with satuated sodium chloride solution
and dried over anhydrous magnesium sulfate. The d.rying agent
was removed by ~iltration and w~shed with 10 ml o~ ethyl
acetate. (The wash sol~ent was combined with ~he original
filtrate). "Skelly~olve B" was then added to the ethyl
acetate to the cloud point (approx~ 10 ml). The mixture
~; was tre~ted with 500 mg o~ activated carbon ("Darko KB")
and filtered. The f~ltrate was diluted with 15 ml of
"Skellysolve B", then seeded with crystals of BL-P2013
free acid. After approx. 3 hour~ at room temperature, the
crystalline precipit~te o~ free ~cld was collected and
dried in vacuo (15 m~n) o~er P205 to obtain 323 mg (~6%)
m.p. slow decomp. o~er 100.
Anal.~;.Calcd. for C8~1oClN~5S: C, 35.89; H9 3,~7; Ns5~23
Cl, 13.25
Found: C~ 35.88, H~ 3.91; N, 5.41; Cl, 13.52
This product was found to be unstable when stored
at 23C. for seven ~ays.
*Trademark
~1'
... .
-41-
Example 15
6a-Bromopenicillanic Acid Sulfoxide
_ -- O
¦ ~r~ ¦(C6~5C~2~C~
2~ ~C02H
MW 800.6~ MW 296.1
,
To 3 1 of methylene chloride was added 300 g
(0.75 mole) of 6a-bromopenicillanic acid N,N'-dibenzyl-
ethylenediamine salt and this suspension was cooled to
5. Then over a 15 min period, with go~d stirring, 1~0
ml of conc. HCl was added dropwlse. me slurr~ was
stirred at 5 for 2 hours. It was then filtered through
a ("Celite") pad of diatomaceous earth and the cake was
washed with 3 x 250 ml of meth~lene chloride.
The combined methylene chloride solutions were
washed with 2 x 500 ml H20 and dried over sodium sulfate
for 15 mln. The sodium sulfate was removed by filtration
and the filtrate evaporated under reduced pressure to approx.
750 ml.
This soIution was cooled to 5 and, wi.th vigorous
stirring, 130 ml of 40% peracetic acid was added dropwise
such that the temperature was maintained at 5 to 12.
The addition was quite exothermic. At the end of the addition,
the slurry was stirred at 5 ~or 2 hours and the product
collected by ~iltration ~d wa~hed with lOO ml of cold ~2
(5) and 100 ml of cold methylene chloride (5). There
was obtained 126 g ~57%) o~ 6a-bromopenicillanic acid
sulfoxide, m.p. 129. The ir ~nd nmr spectra were consistent
for the desixed product.
Anal. Calcd. ~or C8ElOBrN04S: C, 3204~, H, 3.40; N, 4.73.
.. . .
- Fou~d: ~ 3~.3~; H, 3.~5; N, 4 71;
H203 2.18.
,
-42-
~75~
Potassium 6a~romopenicillanate Sul~oxide
O O
Br~" ~ B~, ~
""C02H "~,C02K
MW ~ 4
:
To 3 1 of acetone was added 126 g (0.43 mole)
of 6a-bromopenicillanic acid sulfoxide and 162 ml of 50
by weight potassium 2-ethylhexanoic acid in n-butanol.
After stirring 1 hour at 22, the product was collected
by filtration, was~ed with 2 x 250 ml of acetone and dried.
mere was obtained 127 g (90%) of potassium 6~-bromo-
penicillanate sul~oxide, m.p. 185. The ir and nmr spectra
were consistent for the desired structure.
Anal. Calcd. for C8~rKN04S: C, 28.75; H, 2.71; N~ 4.19.
.
Found: C, 29.0~; H, 2.78; N, 4.04.
-43-
5~
p~Nitrobenz~_6a-Bromo~en~cill~nate Sulfoxide
O O
Br ~ Br ~
~C02K ~ ~02CH ~ No2
MW 431.28
To 1 1 of N,N-dimethylacetamide was added 145
g ~0.4~ mole) potassium 6~-bromopenicillanate sulfoxide,
and, with stirring, there was then added 115 g (0.53 mole)
of p-nitrobenzyl bromide at 22. The mixture was stlrred
at 22 for 20 hours.
The reaction mixture was poured into 3 1 of ~2
and extracted with 3 x I500 ml of ethyl acetate. The
combined eth~l acetate extracts were washed with 2 x 500
ml of 5% aqueous sodium bicarbonate solution and dried
over sodium sulfate for 1/2 hour. The sodium sulfate was
~iltered o~f and the filtrate e~aporated under reduced
pressure to a residue to which 1 1 o~ dlethyl ether was
added causing the product to crystallize. The cr~stals
were collected by filtration, washed with 2 x 100 ml of
diethyl ether and dried to yield 162 g ~87~) of p-nitro-
benzyl 6a-bromopeniclllanate sul~oxide, m.p. 111. m e
ir and nmr spectra were consistent ~or the desired structure.
Anal. Calcd. for C H16BrN O~S: C, 41.78; H, 3.51; N, 6.50.
_ 15 2
Found: C, 41~66; H, 3.45; N, 6.85;
H20, o.69.
-44-
~5~
benz~l 6~-Bromo-2~-chloromethyl-2-methylpenam-
3-carbox~late
~o CE ~ N0 B* ~ ,; ~2Cl
2 2 C02CX ~ N0z
MW 449.71
To 1 1 of p-dioxane was added to 70 g (0.16
mole) of p-nitrobenzyl 6a-bromopenicillanate sulfoxide
followed by 21.2 ml (0.10 mole) of benzoyl chloride and
21.8 ml (0.19 mole) o~ quinoline. m e reaction mixture
was re~luxed ~or 4 hours and then cooIed to 22, poured
into 2500 ml of H20 and extracted into 3 x 800 ml of
ethyl acetate. The combined ethyl acetate extracts were
washed with 300 ml of 5% aqueous sodium bicarbonate
soiution, 300 ml o~ 5% aqueous phosphoric act~~and ~00
ml of ~ 0. m e ethyl acetate solution was dried over
sodium sulfate ~or 1/2 hour and the sodium sulfate was
removed by ~iltration. The filtrate was evaporated
under reduced pressure to a residue which was redissolved
in 1 1. of ethyl acetate and again evaporated under reduced
pressure to a resldue. Then 1 1. of diethyl ether was
added and the product collected by ~iltration to yield
41 g (57%) of p-nitrobenzyl 6a-bromo-2~-chloromethyl-2-
methylpenam-3-carboxylate, m.p 132. The lr and nmr
spectra were consistent for the des~red structure.
~nal Calcd. ~or C EI BrClN 0 S: C, 40 ~ o6; H, 3.14; N, 6.2~.
15 14 2 5
Found: C, 40.62; H, 3.11, N, 6.13.
-45-
~5E~7
p-Nitrobenzyl 6awBromo-2~-chloromethyl-2-methyl~enam-3-
carboxylate Sul~oxide
.. . .. . . . . O
O (~02Cll~r N2 '~ N02
To 1200 ml of methylene chloride was added 51 g
(0.11 mole) of p-nitrobenz~l 6a-bromo-2~-chloromethyl-2-
methlypenam-3-carboxylate followed by 23 g (0.12 mole) of
m-chloro~eroxybenzoic acid. me ~olution was stirred at
22 for 2 hours and evaporated under reduced pressure to a
wet residue. The residue was stirred with 4 1 of diethyl
ether for 1 hour and allowed to stand at 10 for 20 hours.
The product crystallized out ~nd was collected by ~iltration~
washed with 2 x 200 ml of dieth~l ether and dried, yielding
39 g p-nitrobenzyl 6a-bromo-2~-chloromethyl-2-methylpen~m-3-
carboxylate sul~oxide (75~), m.p. 132. m e ir and nmr
spectra were consistent ~or the desired structure.
r C15H14BrClN206S: C, 38.69; H, 3.0~; N 6 07
.
Found: C, 38.98; H, 3.04, N, 5.84;
H20, -35
.
-46
Potassium 2~hloromethyl-2-methylpenam-~-carboxylate
5ul~one_ ~BL-P2013)
.
_ . .... _ . . .
~. - O O
B~ ~ CH2C ~ CH~Cl
O ~2CH~ N2 ~' C02K
BL-P201~
.. . _ .
__ _ MW 305.77 _ --
- -
To 600 ml of H 0 was added 8 g of 30~ Pd on
"Celite" and l~ g (0.19 mole) of sodium bicarbonate.
m en 32 g (o.69 mole) of p-nitrobenzyl 6a-bromo-2~-
chloromethyl-2-methylpenam-3-carboxylate sulfoxide was
dissolved in L~oo ml of eth~l acetate and added to the
aqueous slurry. m e mixture was hydrogenated on a Paar
apparatus at 50 p.~ at 22 for ~ hours. The slurry
was filtered through a thin '1Celite" pad on a s~ntered
glass funnel, the pad was washed with 2 x 50 ml H~0 and
the aqueous layer of the combined filtrate and washings
was separated. The a~ueous layer was washed with 200 ml
of diethyl ether, then was cooled to 5 and~ with stirring,
a solution of 12 g. (.076 mole) of KMnO4 in 200 ml of H20
was added dropwise over a l/2 hour period, keeping the pH
between 7..5 and 8.o by the addition o~ 40~.~ P04. When
the pink color persisted for 5 minutes, no more KMnOl~
solution was added. The reaction mixture was stirred
with a small amount (approx. 50 m4) of sodium bisulfite ~or
l/2 hour, and then the slurr~ was ~iltered through a "Celite"
pad. r~he pad was washed with 2 x 50 ml of H20. The
.. . .. . . . .. . . . ..
~ 5~7
combined filtrate and washings were layered with 500 ml of
ethyl acetate and, with stirring, the pH was adju~ted to
1.5 by the addition of 2 N HCl. The layers were separated
and the aqueous layer was s~turated with sodium sulf&te.
It was reextracted with 2 x 400 ml of ethyl acetate and the
co~bined eth~l acetate e~tracts were dried over sodium
sulfate for 1/2 hour at 5. The sodium sulfate was
removed by filtration and the ~iltrate evaporated under
reduced pressure to a residue. That resldue was dissolved
in 1~0 ml of acetone and 160 ml of diethyl ether a~d ~0
by weight of potassium 2-~thylhexa~oate in n-butanol
was added until the solution was neutral to moist pH paper.
The potassium saIt of BL-P2013 crystallized out, was
collected by filtration, washed with diethyl ether and
dried. Yield 16 g potassium 2~-chloromethyl-2-methylpenam-
3-carbox~late sulfone (BL-P2013) ~76%), m.p. 202~. The
ir and nmr spectra were consistent for the desired structure.
~na~. Calcd. for C8EgCIKN05S: C~ ~1.42; H, 2.97; N, 4.58.
_ Found: C, 31.18, H, 2.98i N, 4.51;
- H20~ 0-93-
.
-~8-
~ ~ 5 ~ ~ 7
Ex~mple 16_
Pivaloylox2~e_h~1 2~-Chloromethyl-2-meth~ nam-3-
carboxylate Sulfon~e (BL-P2024~
. . _
5 0 0 0 .
~S~ C ~ Cl - D~ ~ ~ Cl
~ > '~,CE~
"'~C02K~ Co?cH2ococ(c~3)3
BL-P2024
_ MW 381.8~ . -
_ .. . . ~
To a stirred suspension of 14.6 g (0.0~87 mole~
o~ potassium 2~-chloromethyl-2-methylpenam-~carbox~late
sulfone (BL-P201~ in 200 ml of acetone was added ~ ml o~
a 10~ aqueous solution o~ sodium iodide and the mixture
was brought to reflux on the steam bath. To this reflux-
ing suspension was added 14.8 ml. (0.1 mole) of redistilled
chloromethyl pivalate (bp 34C at 7 mm Hg) all at o~ce.
m e m~xture was stirred at reflux for three hours and then
cooled to room temperature (22 C). The crystalline solids
were collected by filtration, washed with 3 x ~0 ml o~
acetone and the combined filtrates were e~aporated to an
oil under reduced pressure at <22 C. The oil was then
taken up in ~00 ml of ethyl acetate and wa3hed once with
water (200 ml) and once with saturated Na2S01~ solution
(200 ml). The solution was then dried brle~l~ over Na2S0
while being stlrred with 2 g o~ decolori~ing carbon with
cooling (ice bath). A~ter 20 min. the mixture was ~iltered
through a "Celite" pad and the pad washed with 4 x 100 ml
o~ ethyl acetate. The combined ~iltrates were concentrated
under reduced pressure at 22 C to an oil. The oil was
then further concentrated at about 22 C and <1 mm Hg to
remove most of the residual chloromethyl pivalate. The
remaining oil was then tritura~ed twice with 50 ml portions
of n-pentane an~ then left over -the weekend at about 10 C
-49-
~7S13~7
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-pentane. The product waæ
then collected by filtration, washed with diethyl ether-
n-pentane (l:l) then n-pentane and air dried. A~ter
drying under high ~acuum for ~our hours over P205 there
was obt~ined 1~.37 g pivaloyloxymethyl 2~-chloromethyl-
2-methylpen~m-3-carboxylate.sulfone (BL-P2024) about (75%),
m.p. 93 - 95~ C .
: :
,
:
-5~-
~l~L7~ 7
Purification of BL-P2024
Approximately 3 g. o~ crude B~-P2024 (obtained
as described above) was dissolved in 5 ml. o~ ethyl
acetate, placed on a 4 5 x 40 cm col~mn of silica gel
(Mallinckrodt CC-7), and eluted with 4:1 ~/~ C ~ C12-
ethyl acetate. The ~ractions contain~ng a single ~pot
at RfO.84 (TI~ on silica gel plates with 4:1 CE2C12-
ethyl acetate, I2 detection) were combined and concentra
ted ~der reduced pressure to 1.38 g. of a crystalline
solid. A portion of this material (900 mg.) was dis-
sol~ed in 5 ml. of ethyl acetate; the resulting so:Lution
was filtered, diluted almost to the cloud point with
petroleum ether ('ISkell~solve B") and then stored at room
temperature for three days. The cr~stals which formed
were collected b~ filtration3 washed with petroleum ether
and dried to give 560 mg., m.p. 100-101., of purified
BL-P2024.
Anal- Calcd. for C14H20C1~07S C~ 44~03; H~ 5-27; N~ 3-67
Found: C, 44.115 H, 5.o8; N, 3.85.
All temperatures in this application are given
in degrees Centigrade.
5~
-51--
Example 17
Preparation of BL-P2013 Ammonium Salt
1. The free acid of BL-P2013 (250 mg.) dissolved in 20
ml. o~ acetone-methanol (1:1 by volume) was ~iltered to get
a clear solution.
2. Anhydrous ammonium solution was prepared by adding 1 ml.
of ammonium hydroxide ~30%, reagent grade) to 10 ml. of
acetone-methanol (1:1 by volume~ sol~ent and then 1 gm.
of anh~drous magnesium sul~ate was added to that solution
with mild agitation and the mixture was filtered thrQugh
a ~ilter paper~ the ~iltrate was designated "anhydrous
ammonium solution. 11
3. To the filtrate o~ Procedure 1, approximately 2 ml. o~
"anhydrous ~mmonium solution" was gradually added and mixed
well.
~, A 100 ml. portion of diethyl ether was mixed with the
mixture from Procedure ~ to pr~cipitate the ammonium salt
of BL-P20130
5. me white ammonium salt was isolated from the solvent
and washed with 2 port1ons of 50 mlO each o~ dieth~1 ether.
6. The isol~ted powder was dried at 35 C vacuum oven for
overnight.
7. Analytical data were as ~ollows:
Calculated ~ C 33.7; H 4.6; N 9.8;
Found C 33.66; H 4.63; N 10.12; dry by KF
Microscopic Examination: cr~stalline sub~tance
.:~
.,, -
.
~ ~ 75 ~ ~ 7
Example 18
Preparation of Non-hygr~sc~ c Sodium Salt of BL-P2013
1. Dissol~e 50 mg. of the free acid of B~-P2013 in 4 ml.
of acetone-methanol (1:1 by volume) mixture. Filter to
get a clear solution.
2. Prep~re sodium 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 o~ Procedure 1, add the 10 ml. solution
of Proce~ure 2 and mix well.
4. A 10 ml. portion o~ diethyl ether was mixed with the
mixture from Procedure 3 to precipitate the sodium salt of
BL-P201~.:
5. The white salt was immersed in the diekh~l ether for
1-2 hours a~d th~n W2S isolated from the solvent and washed
with 3 portions of 5 ml. ~ach of diethyl ether.
6. The isolated powder was dried at ~0 C vacuum oYen
for o~ernight.
53
~7S~
o o o
S~ CH2Cl Recryst, ~ ~ CH2Cl
f
~c~ ~20-aceton~ F -~c~
'~C02E~H?O C2 2
BL-P2013 (~00 mg.) was dissolved in a mlnimum amount o~
acetone-H~0 (1:1) by volume and diluted with 10 ml. of
acetone, filtered, then diluted with acetone to abcut
25 ml., scratched, and after ~0 minutes the crystalline
hydrate was collected by filtration, washed well with
acetone9 air dried ~nd then vacuum dried at <1 mm. Hg
overnight.
Yield 280 mg.
Anal. C~lcd. for C8HgClNOSK-H20: C, 29.67; H, ~.~9; N, ~.63,
. . . Cl~ 10.94; H20, 5.55.
Found: C, 29,32; H, 3.~2; N, 4.44;
Cl, 11.~1; H20~ 5~9
- s~ -
Example 20
N,NI-Dibenz~lethylened~amine Salt.of BL-P201~
BL-P2013 + 1/2 N,N'-Dibenzylethylened~ami~e diacetate
Recryst. ~ ~ ~ //~ C ~ Cl
Acetone-ether ~ ~ C ~ .
C 02H. C6H5cH2NH-cH2----
. _ . . . . . .
~ o6 mg~ (0.001 ml) of BL-P2013 was dissolved
in 7 ml. H20 and added to a solution of 180 mg. (0.0005 mol)
of N,N'-dibenzylethylenediamine diacetate in 7 ml~ H20.
The mixture was stirred and the salt crystallized and after
stirring approximately 10-15 minutes the salt was collected
by filtration and air ~ried to yield N,N'-dibenzylethylene~
diamine salt of BL-P2013 (300 mg). The material was re-
crystallized by dissolving it in approximately 10 ml. of
boiling acetone and diluting with ether to the cloucl point.
260 mg. of air dried and vacuum dried material was obtained
Anal. Calcd: C, 51.69; H, 5 42; N, 7.5~; Cl, 9.55.
Found: C, 49.39; H, 5~49; N, 7 05; Cl, 8.96;
~2' 1.23 (KF).
:~31.75il~7
Exam~le 21
Chloromethyl E~ter of' BL-P2013
....
O O
\\S~d~CH2Cl + ClCH2-0-S02C
~'CH3 + (CE3CH2CE2CH2 )4~ES04
0~ "COOK ~ 3 KHC03
t35.7)
I CH~C12
~ H2
O O
~ ~2Cl
O C-ocH
O
(316 ~ 17 )
... . .. , , ,, . ,,, , , , " . ... .. . .. . ..
...
.
-56-
~7S~7
To a vigoxously stirred mixture of 15.25 g
(0.05 mol) of BL-P2013 (5~, 15 g. (0.15 mol) KHC03 and 1.7
g (0.005 mol) o~ tetrabutylammonium hydrogen sulfate
(Aldrich Chem. Co.) in a mixture of 5V ml. water and
50 ml. CH2C12 there was added dropw~se a solution of 9.5
g ~0.0575 mol) of ClC~2-0-S02Cl in 40 ml. CH2C12. m e
temperature rose to 26 C. and after the addition (wh~ch
took about 15 minutes) the mixture was stir~ed another 30
minutes. Because the product cr~stallized out more CH2C12
(about ~00 ml.) was added to obtain a solution~ The
separated CH~C12 layer and a 50 ml. C ~ C12 wash were com-
bined, dried over MgS04 with stirring and 2 g of decoloriz-
ing carbon ('tDarco KR") was added. ~ter about 30 minutes
the mixture was ~iltered, concentrated to about 50 ml. and
isoprop~l alcohol (150 ml.) was added~ The rest o~ the
CH2C12 was then removed under reduced pressure. The
resulting crystalline precipitate was col~ected by filtra-
tion, washed well with isopropyl alcohol and air-dried.
After vacuum drying at less than 1 mm. Hg there was obtained
8.5 g of chloromethyl 2~-chloromethyl-2-methylpenam-3-
carboxylate sulfo~e (73. M.p. 116 (dec., dar~ens above 100C).
Anal. Calcd. ~or CgHllC12N05S C, ~4.18i H, 3.51; N, 4.~3;
Cl, 22.4~.
Found: C, 34.16; H, 3.45; N~ 4.47;
~ Cl, 22.46, ~ 0, 0.33 ~KF)~
~Bstimated puxlty in the 90-95~ range.
~s~
Iod ~ r of BL-P2013
H2C1 ~ CH2Cl
Na I ~ ~ oC~3
To a stirred mixture o~ 5 g. (0.0159 mol) of
the chloromethyl ester of BL-P2013 (7) in 2~ ml. acetone
was added ~ g. (0.02 mol) of sodium iodide. me resulting
slurry was stirred for 17 hours and then cooled to about
0C. Two drops o~ saturated aqueous KHC0~ were added
and the mixture was slowly diluted d~opwise with water
over ten mi~utes until 50 ml. had been added. m e
slurry underwent a sudden color change from yellow to
grey to purple to black and therefore the crystals were
immedi~tely collected by ~iltration and washed with cold
acetone-water (1:2) and then isopropyl alcohol (3 x 10 ml)~
then diethyl ether and finally n-pentane and air-dried
to yield 5.55 g. (91% ~leld) of the iodo~ethyl ester of
BL-P2013 (8). M.p. 118 - 119C. with decomposition.
Purity estimated at about 90%.
_. . .
6-[(R)-2-Amino-2-phe~ylacetaml~o]-~,3-dimethyl-7-oxo-4_
thia-l-aZabicycloc ~ thyl
2~-chlorometh~1-2-a-methylpenam-~a-carboxylate sul~one (11)
, .... . . _ . ,
58-
~1~5~
8~ - C~f~-Co-~'COOK
C~
OCH~Patent 3, ~16, 247 )
I
~ ~ '
~=~ CH~ -
~-CH--CO--~ ~ ~CH3
~N O C=O
H~C--If H
H C~C~D
O~H3 0
O O
o~, ;H2
o
... . ' I ~
~j
.~
-59-
~a~75~7
C~L
CH C0 - NH ~ C
NH~ o C~O
o
:
::
~ : ~ ,r~ 'fs' l~ ~
:
To a stirred mixture cooled in an ice-bath of
5.~6 ~ (0.01 mol) of the indlcated Dane salt of amplcillin
9 (which was sol~ated with one molecule of isopropyl
alcohol) in 60 ml. acetone there was added 4.o8 g. (0.01 ml)
o~ the iodomethyl ester of BI~P2013 (8) and the resulting
nearly clear solution was stirred for f~ve hours w~th ~he
ice-bath remo~ed a~ter 30 minutes. m en most of the~ acetone
was removed in vacuum on the roto-~ap and the resulting
concentrated solution was dlssolved in 200 ml. co~d ethyl
acetate which was then washed with 2 x 50 ml. ice cold water
and 2 x 100 ml. saturated aqueous Na2S04. me ethyl acetate
so.lution was then dried over Na2S04, filtered and most of
the ethyl acetate was removed in vacuo on the roto-vap~
The residue was triturated with 2 x 200 ml. dry diethyl
ether and the resulting solids were collected by ~iltration
to give 5.5 g. of 10 as a pinkish powder. This powder was
..,
.
:
:.
- -60-
D7
stirred in a mixture of 50 ml. water, 50 ml. n-butanol
and 20 ml. ethyl acetate while 6N HC1 was added dropwise
to pH 2.5. Then occasionally a drop or two of HCl was
added to keep the pH at 2.2 - 2.5 over 45 minutes. ~hen
the pX no longer drifted upward there was added to this
mixture 100 ml. diethyl ether with good stirring~ The
aqueous phase was separated an~ 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 layer was then stirred vigorously
under a layer of 100 ml. 2-butanone (meth~l ethyl ketone)
while Na2S04 was added to saturate the aqueous layer~ The
2-butanone l~yer was separated~ dried over Na2S0~
for 30 minutes in an ice-bath, 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~3.2.0]heptane-2-
carbonylmethyl 2~-chloromethyl-2-a-methylpenam-~a-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-phenylace-
tamldo]3,3 dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]-
heptane-2-carbonyloxymethyl 2~-chloromethyl-2-a-methyl-
penam-3a-carboxylate sulfone.
~61--
~l~S~
Exam~le 22
Improved Synthesis of BL P2013
This procedure simpli~ies production of
BL-P201~ by elim-lnating the previous use of catalytic
reduction .
Step 1
Br 1'
"/, , S~,<
o ~ COO~CX2CC ~ + Dicyclohexylcarbodiimide
1 Pyridine
l C~2C12
-
O
Br~ ~ 2
N
O C-OC~2CC}~
o
( See page 633 of Cephalosporins and Penicillins, edited by
Edwin H. Flynn, Academic Press, New York, 1972)
6a-Bro~openicillanic acid sulfoxide ( 1 ) (30 g.,
0.1 mol) was dissolved in 1 1. dry CH~C12 followed by
the addition of 16.2 ml. (0.2 mol) pyridine and 29.8 g.
~0.2 mol) trichloroethanol. ~hen 20 g. (0.1 mol) o~
dicyclohexylcarbodiim~de was added and the mixture was
3tirred at 22 for 16 hours. Dicyclohexylurea began
~L~75~7
to precipitate out; at the end it was removed by filtration.
The ~iltrate was washed with 200 ml. o~ 5~ aqueous sodium
bicarbonate, 200 ml. of 10% phosphoric acid and 100 ml.
of saturated aqueous sodium sulfate. m e organic phase was
dried over sodium sulfate at 5C. ~or 30 minutes, filtered
and evaporated to an oil. Dlethyl ether was added and,
with scratching, the product 2 crystallized out (27 g~,
6~ yield).
s~e~ ?
2 ~ /=\ o
- ~ CC1
¦ ~u~noline
Br C ~ Cl
~ -''~CH3 ~ :
0~ C - OC~2CC1~5
o
Compound 2 (26~5 g., 0.062 mol) was dlssolved ln
500 ml. p-d~o~ane and there was added 8~5 ml. (o.o78 mole)
benzoyl chloride and 8.75 ml. (o.o78 mole) quinoline. m e
solution was reM uxed ~or four hours and -then poured into
1100 ml. water and the product ~ was extracted into
2 x 400 ml. ethyl acetate The e~hyl acetate extracts
were combined, successively washed with 200 ml. 5~
aqueous sodium bicarbonate, 200 ml. 5~ phosphoric acid
and 200 ml~ saturated aqueous sodium.sul~ate, driea over
sodlum sul*ate at 5C. for thirty minutes and evaporated
to an oll ~) which was used "as ls~' ~or the next reaction.
-63-
Step 3
3 ~ KMnO ~H O
in glacial
acetic acid
~: \ /
; O O
Br \\ ~ ~CH2Cl
C ~ 4
O C-OCH2CC13
~: O
~: ~
Compound 3 obtained in the previous step was
dissolved in 1 1. glacial acetic acid and, w~th stirring
at 2?C., a sa~urated aqueous solution o~ KMnOI~ was
added dropwise until a pink color persisted (that is~
a drop placed on a piece of filter paper gave a pink
coloration). Then with cooling 30% H202 was added
dropwise until a clear solution wa~ obtained; some
white precipitate was present. The solution was
poured into 2.5 1. water and the product 4 was extracted
into ~ x 500 ml. ethyl acetate. The ethyl acetate w~s
washed wlth 5% aqueous sodium b~carbonate until neutral
(that is~ no more bubbling upon addition), dried over
sodium sul~ate and e~aporated to leave 4 as the residue.
.~ ~
-64-
~3L75~1~7
It was le~t at 10Co for one day and then triturated
with "Skellysolve B'l to yield 9.1 g. solid 4. The
yield was 28% of theory ~or steps 2 and ~ combined~
Step 4
Zn in Acetic Acid
KEH
\ ~
,U~C~2Cl 5 (III,-P2013)
~; ~
o COOK
(See U. S. Patent 4,164,497)
Zinc dust (3.75 g.) was slurried in 5 ml. glacial
acetic acid and cooled to 5C. To this ~xture there was
added a solution of 4 (3 g.; 0.0057 mole) in 15 ml.
dimethylformamide and the result~ng slurry was stirred
at 5 for 2~5 hours.
The zinc was then filtered off and the pa:Le
yellow solution was poured lnto 80 ml. of 5% aqueous
hydrochlorlc acid. That mixture was extracted with ~ x 25
ml. ethyl acetate. The combined ethyl acetate extracts were
extracted wi~h 3 x 20 ml. of 5% a~ueous sodium bicarbonate,
sa~in4 the ethyl acetate phase after separation.
--65--
~l75~7
The bicarbonate extracts were combined~ placed
under a layer o~ ethyl acetate~ adjusted to pH 1.5 by
the addition of 2N ECl and saturated with sodium sulfate.
The ethyl acetate was separated and the aqueous phase
was extracted with 2 x ~0 ml. ethyl acetate.
All of the ethyl acetate phases above were
combined, dried over sodium sulfate and evaporated to an
oil (which was the free acid form o~ BL-P2013) which was
dissolved ~n about 20 ml. acetone to which 20 ml. cliethyl
ether was then added. Then 50% potassium 2-eth~lhexanoate
(KEH) in dry n-butanol was added to neutrality. I~le
product 5 (~L-P201~ crystallized out. After stirring
0.5 hour at 22 it was collected by filtration to yield
650 mgm. o~ 5 (37% yield).
A 50 mgm. sample o~ 5 wa~ dissolved in 0.5 ml.
water and 20 mgm. N,N'-dibenzylethylenediamine (DBED)
diacetate was added. The DBED salt o~ 5 crystallized out,
was collec~ed by ~iltration, washed with water and dried
over P205 under vacuum to yield N,N9-dibenzylethylenediamine
2~-chloromethyl-2-methylpe~am-3-carboxylate sulfone (DBED
salt of ~ree acid 5~.
Another s~mple of 5 (450 mgm.) was dissol~ed in
3 ml. water ~o which was added a solution o~ 270 mgm DBED
diacetate in 2 ml. H20. With scratching the DBED salt of
5 crystallized out (4~0 mgm~). Recr~stallization ~rom
about 5 ml. boiling acetone yielded 270 mgm.
, ~
--66-
Example 23
6-[~R)-2-Amino-2-p-hydroxyphenylacetamido]-3~3-d~methyl-7
oxo-4-thia-1-aæabicyclo[3.2.0]heptane-2-carbonylo~ymethyl
2~-chloromethyl-2-a-methylpen~m-3a-carboxylate sulfone
having the formula
HO ~CH--CO NH ~CH3
NH2 0 C~O
I
O
.. .`', O
i8 produced by substituting the corresponding Dane salt o~
amoxicillin for the ampicillln used in the procedure of
Example 21.
--67--
S~
BIOLOGICAL DATA
~ he product of Example 1, Compound 5, having
the struc:ture
(~O~CM2Cl
C0
.
will be referred to below as Bl.-P2013.
Al~ough at best a Yery s~eak antibacterial agent
itself, BL-P2013 inhibits ~-lactamases and protected
ceforanide and amoxic:illin from destruction by ~-lactamase-
~~ producing bacteria in vitro and in vivo.when used in
:~ combination with those two agent~.
,~,
--68--
75~7
Table 1
Antibacter al Activity of_ the New Sulfone
.. _ _ .... . _ .
MIC (mcg/ml )
. . ~ .
Organism
BL--P2 013 Ampicill~n
-- ~ . . ... _ _ ~ ....... _ .
~_ ~ . . ,
S . pneumoniae A-9585 16 O . 004
p; .. pyogenes A-960ii4 _ 63 O - _
ts. aureus P~-9537 _ >125 0.16
, ~ . _ ~ .
S. aureus +
50% serum A-953? ~125 0 . 06
. _ . . _ _ .. .
S~. aureus Pen-
Res A-9606 _>125 ~125
S.. aur~us Meth-
es ~15097 >125 125
S. faecaiis A20688 >125 0.13
E. coli A15119 ~12 5 ~ --- - - A ~ _ _ _
_ ~ _ _ , _ __ _ ~___ _ _ _ _ _
E. coli A~0341--.1 ~125 ~125
R. pneumoniae A15130 ~125 125
K. pneumoniaé A2 04 68 >12 5 > 12 5
P. mirabilis ~A-9900 ~125 0.13
P. vulgarls A21559 -~125 ~125 .
_ . .... _ _ . _ ,.. _ .. , _ - I
P. rnorganii A15153 >125 >125
P. rettgeri A21203 ~12 5 ~ ~ l
S. marcesce~s A20019 >125 16: ¦
., . ._ _ _ . , __, . . . _ ._ _ ._
E. cloacae A-9659 >125 63
. E. cloacae A-9656 >125 >125
. . . . _ _ _ . . ~
P. a~ruginosaA-9843A ~125 ~lX5
. E'. aerugi-wsa AZ~Z~ 2S ~125 ~ ~ .
--69--
~75~7
Tabl e 2
Anti-Bacteroides Activity of Ceforanidle and Amoxicillin
Alone and in Combination with BL-P2013
_ _ . , ~ l
eta MIC ~mc:g/ml) ~
Organism lact- ~ _ _ ~ .
. amasranide~B~-P~013;~013 ~BLP2013c:illi
r
B . f ragil is
A2191 ~ 63 2 >125 2 8
A22053 ~ 32 4 63 2 8
A2202 ~ 32 2 32 2 4
A2187 + 32 4 63 2 8
A22534 ~>125 32 125 16 >125
A22 697 + 63 8 63 2 8
A2269 ~ 63 4 63 2 16
P.22~94 ~ 125 16 63 2 16
~2269 +~125 _16_ 32 4 1~5
A2269 +~125 _32 63 E~ >125
A2253 l>125 _32 32 32 >125
A2253 +~125 32 125 32 >125
A2279 ~~125 8 32 4 125
A2279 t~ 3Z 4 32 ? 8
A2279 ~ 32 4 32 2 8
A2279 ~ 63 4 32 4 16
A2279 ~ 63 4 63 2 16
A2279 ~ 32 4 63 2 8
B. theta-
iotaomic-
ron A2227 -~- 125 4 63 2 16
A2227 ~ 125 8 63 4 16
__ . _ _._ ____ _ ___.
... .
-70-
7551~7
Table 2 - cont'd.
Anti-Bacteroides Activity of Ceforanide and Amoxicillin
Alone and in Combination with BL-P2013
_ _ MIC (mcg/mI)*~
Beta
Organism lact~ Ff o- Ceforanide B Amoxicillin Amoxi-
: amasranide+BL-P2013 2013~BL-P2013 cillin
~ _ , ,, _. ~
Bacteroides
species
A2Q93 ~32 4 32 2 8
A2195 +63 4 32 2 16
A2092 ~63 4 32 2 16
A2195 ~63 lfi 63 2 16
A2093 ~63 16 63 _4_ 8
: A2093 ~125 8 125 4 32
: jA2093 ~~3 ~ 32 2 1~
.~ --A2~927- _0.5 1 63 0.13 0.13
~09~ _ 2 2 125 0.13 0.13
good synergism
--- marginal synergism
.. .... .
~Minimum inhi~itory concen~ration ~MIC) determined by the
agar dilutio~ method usi~g 50X dilutions of 24 hour cultures
as inocula dispensed by the Steer's inoculatorO Assay
mcdium composed o~ Brucella Agar plus 5% lakëd sheep blo~d
and 10 mcg/ml vitamin K~
--71--
~7S~
Tabie 3
Therapeutic Effica~:y of Amoxicillin in Combination with
BL-P2013 in Mice Exeer~mentall~v Infected with a_Beta-
Lactarnase Strain of Staphylococcus aureus
_ Chaïlenge 5~7~eat:ment (mg/kg)~ - ------
Organism (No. of ~moxicillin ~A` BL-P2013 tB) A+B (1:1)
_ rganisms~ IM PO IM PO IM_ PO
:~S. aureus 5 x 10 >800 ~800~50 ~200 6.3 44
: A--9606 5 x 10 >800 >800>50 >200 19~7
: ~ ~ ~50 _ 9.6 -
:~
Treatment schedule: Drugs administered 0 and 2 hours post-
inf ection .:
:
.
.
-72
~5~7
. .... ... ,..... .
~: w ~ ~ _ :a
. t~ ~ t~1 0
~ ~ ~ o ~ ~
U~ O O 1:; N
. ~ j_. :~ 0 1-'
~ P ~ P- !~ 9~
~a .. -- ~D
It _, ., ,, ,, ... ,, ~ ~ U~
~ h~ ~ a ~ ~h
~ O O C:~ O O O ~ o . ,~ o
O O O O ~ O
P~ . . _ o
~ ~. _
~ ~ ~ ~_ l_ ~>
pl ~ N ~J ~ ~ )~ O
~ d ~ ~ Ul
~D ~ ~ ~ ~n ~. ~ w
O ~ W ~ Ul ~ W
H) $- ~ ~ . ~O O
0~ ~ O
~D ~ ~ r~
O ' ~ t~ ~ ~ ~D
U~ CO ~ ~I ~ ~ H~ r~
r~ ~ ~r~o ~ ~ ~a
~: O : ~ ~J ~_ ~O~ ~ O I
: : ~ ~ I_ ~ 'C
O A V ~ ~ 1-~
~:: c~ .~ 3 o ~ O ~
w ~ ~ 3 0
l~ AU~ _, t~ ~
~t U~ L~ Ul ~ ~ 1_ ~ 1- 3
u~ . .. . ~ .. ~ ~ ~'D
~- ~ Ul Ct~ ~ ~n ~ ~
O ~:
N ~ ~ ~ ~ O I_ t~
. . . ~ . U~ ~1
U~ 133 N 0~ C~ CO O llr
. (D
. _
. . 31':C .
o~ ~I ~ n ~
O o o O I o :1 ~D I
. __ ~ _ .
PJ ~ O
I . I I I I ., 1.-
9 ~D ~O ~ P~ U~
a~ o~ ~ a
oG o~ ~O O~: os: ~1 ~
a~ n ~ a~ n 1~ ~ ~C
~~ ~-
u~ ~o ~ u~
- . . . ,_ .
~ .
s~
--73-- .
P~
.
~ o ~a
8 ~ c o o u~ o i,
. - _ ____ IU O
oo q~
o o ~ ~ V
; o V ....... U~ ~ ~ o
V ~o~ 5~
~ a~ ~ U ~
u~ P~ ~ qC
~ ¢. ~o ~ ~ ~9 ~ e
. ~ ~ ~ ~u ol ~1
:~ . ~ ~ o ~I U
~_
P: ~ ~ D~ ~ ~
o ~ U~ ~ ~ O
._ __ _ _ 3
~ ~ ~ ~ c~l ~
o ~ ~ _ _ ~ ~ ~, , e
cq o
. ~
--74--
. _ . . . . .. , -
U~ O tq ~3
. ~ . ~ . ,_ . ~o . ~S :C
_ o o
n ~ ~ o ~ o
~ O ~ O ~ ~ ~ p~
W Y ~ W
i~- ~....... ~ ~1 3
. , ~ ol Ul ~
. . _ ,_
~ a~ Ul Ul ~ ~ O o2: ~ ~
X X X X X ~C X X X X X .sl Pl ~ ~b
~ ~ ~ ~ ~ ~ o i- ~ n
O O O O O O O O O O O ~ ~h tD ~
~n Ul U~ ~ ~ C~ CO ~ CO ~D C~ ~ :~ ~D a
o ~
~0 ~ (D O
. ... ~
V V V V V V V V V ~ V ~ 1'- :3
.a ~ ~ ~ ~ CD ~ a~ :~ O ~ O
~ o o c~ o o o o o o ~a X ~ X
O O O C~ O O O O C~ O O ~ I.~G
n t a
_ _ _ _ ~ ~ t~ ~
. 1_ ~ 1- ~3
_ V ;~ ~ 3 1~- ~_
, J ~ Ul _J ~ IV ~ C~
.. .. ., .. ~ ~ o o ~ t
.. .. c~~ X ~3 ~D O
~ ~ v Y n
.. ~n ~ ~
O ~ ~ O ~J-
_
tD
~_ Ul ~ 1- '`I W ~ ~ 3 ~1. ~t.
cow ~ ~ ~ :1 0
., ,. ~o..... " .. .... .. .. ~ ~ ~q
~ w w ~ ~n ~ ~ ~n w ~ ~ I_
. Ul ~ ~ ~ ~ Q U~
: ~ ul ~n I_ ~ ~ ~
t_ .P ~ O,P ~ W
.G~ t~ o~ ~ w ~ ~_
a~ ~ N ~ ~ ~ Ul t~
I_ W U~ O ~~ W I-h N
I~ W C:l
o ~ ~ cn o 0 ~_ Icn 1~
. o . . . .
O O ~ ~ ~ O~ ~
~ . C ~
w a~ ~ ~ I_ ~D
~, ~ ~ ~n .. C 3:
~n ~ . i~ ~ ~ ~
~ Vl O~ O ~tD
. . _ __ _ _ _ 1
. V V V V V V V V V V
~ ~ ~ ~ ~ ~ ~ 3 ~ o t
O S:~ ~ O O ~ ~ ~ O O O )- I O
O O O O O O .P O O Q O 1~ td
--75--
~75~
___~_
o ~ ~3 n
a~ ~ x ~ O
_ n ~ ra
~ ~D ~n o ~ ~ ~ :
n, ~ ~n ~ ~ ~
~D rt ~ ~ ~ ~D
n
~ tn . __ .__ ~ i-~
c o ~ _ ~ n
o~ O Z ~ I_ ~
~t O ~ ~ ~h
C X X ~C~ D~ ~b
~b 1~ 3 0 ~- :1 n
n~ .. o o o~ ~b ~D ~ P~
. m Ul ~n 3 ~ o
1~. ~ rn ~D ~t
: ~ ~ ~ . ~ o
:~ ~q ~ , . , ~, ~
,~ ~ ~, v v ~: ~ x
~D O o O O ~ t)
o o c:~ ~ I t~
It .- .. .. .- ~ ~ O
ID ~ a ~ I_ t~ tD ~ 3
a. . ,.. ~ o
O ~_ D~
P~ ~ g ~n ~'
0~ ~ ~ ,3~ ~ 'o'
o ~n ~ w` ~ ~ ~t~
.~t~
I~ h~H~ N
C Ul b N o
le 3
~ tD . .
, ,_, _ . ~b
! ~
.. ~ ~1~_ In
~, .
.'75~
--76--
_
, . ~ ~r O o U~ O ~ V
al ~ ~ ~ ~ 0
~ C ~
~ ~1 ~ ~ ~` _~ ~
v 0 ~ o _1u~ X 3
S C E r~ ~ ~ ~ N
c ~ ~ ¦ c c ~ ~ . . E 3
; O J~ ~I N U~ 0 Cl
Ga ¦ h U ~ p C
Z 1-~ ~ o I ~ ¦ ~1 0
a I ~ D G ~ --I --I A ~ ~ C O
_ ~
O ~ GS ~ ~ O O U~ ~ ~ C C .8
__ ~ O ~o U"~
O ~ ,~ tl~ ~ v o C r
~ O ~ o o o o ~ O 1:~ S
.C~ ~x x ~c x ac ~e ~
w ~ 3 c~-- ~ D ~ c G
. ~ ~ ~ C
a ~ 4 ~ S ~ --~ 5
--77
5~7
. _ . . . ~ . .
tII t~ W lll t~ tc~ Q
O tl 1~ tl t-l t' tl
~d ~d
o ~ r~ r~ ~ n) r~) o
U~ ~ O O O O O O
~ 1- 1- 1-
c~ ~ a~
. _ _ ~ _ ._ -,_...... O
O ~ ~ O Ul ~ ~qO~ ~
O O Ul O O Ul ~D O
~q 6~
. n ~ ~ _, ~D
o ~o ~ ~ _ ____ . _ _. ~
O ~~ _~ ~_ ~ ~ ~ ~ IJ
3 ~ 1- 1- ~ ~ I- O
. . . . . ~ o
D I_ CO I- ~ ~ ~ ~' R-
~D ~D I ~1 I n~ I IJ I \~J I r~ I I-- ~1
~' ~' 1'- ~' ~' ~'
I . ~ . ~ . ~ ~ C~ 0~ . ~ ~D
O ~Jl ~ Q ~ _~ ~ ~O 0:) ~1 ':
O C~ ~_ ~_ _~_ ~-_ ~ _. I (D
~ ~ ~ ~ ._ ~ _~ ~ ~
~ ~ I_ r~ r~ ~ lD O
~ o . ~ ~ n~ . 1- ~ ~ I ~ I n) ~ G
~ cr.- ~_ . I~) ~ ~D ~ Ul I~ ~ O c~
~5 ~n ~ 1-~ ~D ~\>J ~ ~ ~ t~l ~
. . . . _~ ~ ~ Ç
a) ~ 1~ C~ 1- 5~ 0 E~ Il) t-~
_, _~ ~ _~ C~' IJ IJ' I~ ~D
~-- .. _ _ . _ lD O ~ O
E31 ~1 ~ O 11~ ~ O R ~ ~1
~ ~1_ COI- ~ n~ r~ a~o
I ~- ~ ~ ~ ~ ~ O ~b
~D n~ 1- o r~ ~n~n 1_~
C~ C~ ~ ~ ~1 \~J
~_ _~ ~_ _, _~ ~_ :~
~ _ _ ~-A ~5 EQ~ ~
n) IJ ~ ~ a~ I-
. ~ I~ . . . ,. .
Ul~ ~ ~ ~n~ ~ CJ~ ~ O ~`
I ~ I Ul l I ~)J ~ 1-
~rl IU 1~ 1~ ~ ~
1- ~ W
_, ~_ ~ tl
~ , ... __.
~ O O î~ O O ~0
. ~ . ,.1 . . ,~. ~ O ~ I'
ot~ cr~ I_ ~ a~ 1
4~ I 1' I ~Jl I Ot) O
. 1~ r\~ 1~
~n
~_ ~_ _~ _,
_ _ _ ..... _ _ .
_ ~ ~ _ ~ ~ .
O O O O O ~J
n~ ~n o ~ ~ o ~ ~ o
~ ~"CO 0tX)
_ _~ _ _ _ ~.
--78--
~7~i~iO7
:~ q ~ ~ ~ ~ t, ~ ki~- _- _ .
D o N ~ 10 10 N ~ O
s~ O O O O O O
~o~ ~ ~ ~ r~ ~ n~, ~
_ . ,
IJ 1- 3 t1
0~ct ~ O O 01 O O N ~?
W :5 ~ Oq
i- o ~ ~ - _ _ ... ,.. _ _~___
~a i' ~ ~ ~ ~ ~ ,_
r~) ~n o r~
D ID _J ~n r~ ~n C~ CJ~ IJ
O ~D ~ I ~ ~ r I ~ I ~ I ~ s ~ u
o ~ a~ ~1~ u~ ~ ~J ~
~Jl O . ~ ~--o ~ I~ ~1 ~1 . _J
" ~q ul ~:) a~ ~ ~n
._ _ . ~ .
,_ ~ _~
~D ~ ~_ n~ ~n I_
~ . ~ . ~ . ~ . ~ ~ r- t
`Q. ~ ~ r-~ n~ 1'~ ~ o~-
I ~1 ~ a~ I ~1 I ~ I u~ O ~ ~
r~ 1_ ~ ~ ~ ~3
~_ _~ . ~ ~?l ~_ :S ~D
~ ~ ~n ~ ~ ~ ~
' ~ 80 _ ~_ ~ ~ ~ ~Q o ~
1~ ~ ~1 ~J 1~ ~ r~ O ~ P- ~i .
~t ~D ~ ~ ~ C~50 iO~ ~*r ~1~ ~ O, ~ ~
~5 ~51~1 ~1 i~ N i--~ CCI ~O ~D ~l~J ~ ~
. ~ ~D _3 CO ~ C~ C~
- 1- ~ O -
o ~n~ ~ ) ~ ~ ~O ~ ~O ~
I ~ I 1~ I ~ I ~ I CJ~ I C~ c~ El
I_ p~ ~
CJ~ C:5~ ~ ~ i_ 03 O
_ _ . , l _
_~ _ _ ~_
_~ O O 1~ 1_1 A
~ ' a~' ~ i~ ~ ) I'
~ ~I~n I~ ~0 ~ ~ O
CS~ ~ 0 ~J
. - . _ _ --, .
_ ~ . ~ ~
O O ~ ~ O V
~ O CO~I ;O 1' O IJ
~ 'o l ~ ~
.
--79--
~75~
D q C~ ~ ~ ~ ~ ~ W ~
t~ t~ t~ t~ t~ t~ o
~ ~ r~ ~ ~ r~
o ~ ~ ~ ~ ~ ~ o ,~
__
D O ~n ~ lo ~n r,~ . ~,tl l
~q
O o tD ~ _ .~ _ _ _ _ - -.. ,
i- ~ . ..
_~ ~ ~ _ ,_ ~
~ ~ ~ ~ r,~ . ~ IJ -
IJ-J ~ ~O OD C~ cr~ i- l- Ul
~ ~ ~ ~ ~O ~ ~t ~O Co ~ (J~ 1- ~
. I~ ~1 Cl~ ~
N _ _ _ _ _ .
:D ~ ,_ __
Q ~ ~ r~ ~ ~ I~ ~
~ ~ ~ ~ ~ r ~ 1_ 1~ Ul ~n N O
::s cc IJ_J -`JCO ~O~Jl I-~Jt ~n ~ ~ ~3
o n~ co ~ ~ ~ ~ ~
_ ~ ~ _~ _~ _~ ~D O
_ _ . U~ O
S~.
ct ~ ~ ~ I_ \~1 N O ~ c~
~0 ~D ~ ~n . ~.~J . \~ o~ I ~1 . ,., ~ CJ~ 1~ a~
~D ~O~ CO' ~ I~~ ~O~ ~ ~t
. _~ ~ N~ I~ ~ ~)J
~Jl N ~ D ~
. _ _ . ~ 3
O ~ I~ l_ ~ ~_ O
I ~ . ~ ~ n~ ~ r- . ~ . ,~ ~ ~ ~
. ~ ~ ,_. ~ ~ ~ 3`
I ~ I ~n I n~ I Y I n~ ~ I_
_ ~ co ~ r~ P~ ~_
CO
. _ _ _
. r~ ~ ~'r~ _ c~ ~
oo ~ ~ r ~ r~ ~o . r~
~n ~ ;o rv ~ ~n n~ . o
C~ _ _, ~ ~_ .
. __ . ~ . _ _ _
~- O î- O O .
CON J ~ . . ~ N ~ ~ ~1
~_~ roo -J O co~n 1 i~ O
~o ~ .
_ __ _ _ _
_
~ . . .
- --80--
5~
The compounds of the present invPntion are thus
useful, given orally and parenterally, for enhanc ~ g the
effecti~eness of ~-lactam antibiotics against B-lactamase
producing bacteria. On a weight basis, the dosage is from
one-fifth to five times, and preferably e~al to, t}lat of
the ~-lactam antibiotics. As an example, the compoun~s o
the present invention as shown above when used in a 1:1
ratio markedly improved the activity of ceforanide and
amoxicillin against ~-lactamase producing strains o~
anaervbi~ Bacteroides such as B. fragilis, ~ thetaiotaomicron
and other species of that genus and also against resistant
~ aureus~ The compounds vf the present
in~ention axe given either in admixture with or concomitantly
with the ~-lactam antibiotic w~ th the dosage within the
indicated ratio tc the known and customary dosage oiE the
anti~iotic ~.
Thus, the ability of the compounds of the present
invention to enhance the effectiveness of a ~-lactam anti-
biotic against certain ~lactamase-producing bacteria makes
the~ valua~le for co-administration with certain ~-lactam
antibiotics in the treatment of bacterial infec~ions in
mammals, particularly man. In the treatment of a bacterial
infection, a compound of the present invention can be co-
mingled with the B-lac~am antibiotic, and the two agents
thereby administ~red simultaneously. ~lternatively, a
compound of the present invention can he administ.ered as
a separate agent during a courYe of treatment with a
~-lactam antibioticO
-81-
~75~qa 7
When using a compound of the present invention
or salt thereof~ to enhance the antibact:erial activity of
a ~-lactam antibiotic, it can be administered alone, or
preferably~ in formulation with s~andard pharmaceutical
carriers and diluents. A compound of the present invention
which is in the acid form or.as a pharmaceutically-
acceptable salt thereof, can be administered orally or
parenterally; a compound o the present invention in the
form of an ester which is readily hydrolyzable in vivo, is
best administexed orally. Parenteral administration includes
i~tramuscular, subcutaneous, intraperitoneal and int;ra-
venous adminis~ration.
When a compo~na of the present invention is used
in the presence of a ~a~rIer or dilue~t, said carrier or
diluent is chosen o~ the basis of the intended mode of
administration. For example, when consid~ring the oral
mode of:administration, the c~mpound can be used Ln the
form of tablets,.capsules, 102enges, troches, Eowders,
5yrups, elixirs~ aqueous solutions and suspensions, and the
like, in accordance with standard pharmaceutic21 practice.
_ _ .. . ... ... ., .. ., .. ._ .. s ~ . .. .. ..
The proportional ratio of active ingredients to carr:ier wi}l
naturally depend on the chemical nature, solubility,
stability and potency o~ the active ingredients, as well as
the dosage contemplated. However, these pharmaceutical
compositions will likely con~ain from abou~ 5% ~o about
80~ o~ carrier. In the case o~ tablets for oral use,
carriers which are commonly used include lac~ose, sodium
citrate and salts o phosphoric acid. Various disintegrants
such as starch, and lubricating agents, such as magnesium
stearate, sodium lauryl sulfate and talc, are commonly
use~ in tablets. For oral administration in capsule form,
useful diluents are lac~ose and high molecular weight
.~
-82
polyethylene glycols. When aqueous suspensions are
required for o~al use, the active ingredients are combined
with emulsifying and suspending agents. If desired, certain
sweetening and/or flavoring agents can be added. For
.
parenteral administration, which includPs intramuscular~
intraperitoneal, subcutaneous and intravenous use, sterile
solutions of the active ingredients are usually prepared~
and the pH or the solutions are suitably adjusted and
buffered. For intravenous use, the total concentration of
solutes should be controlled to render the preparation
isotonic.
Although the prescribing physician will ultimately
decide the dosage to be used in a human subject, the ratîo
of ~he daily dosages of a compound of the present inv~ntion,
or salt thereof r and the ~-lactam antibiotic will normally
be in the range from about 1:5 to 5:1, and preferably about
l:1. Additionally, the daily oral dosage of each component
will normally be in the range from a~out lO to about 200 mg.
per kilogram of body weight and the daily parenteral dosage
of each component will normally be about lO to about 100 mg.
per kilogram of body weight. These figures ar~ illustrative
only, however, and in somë cases it may be nécessary to
use dosages outside these limits~
This invention is capable of industrial
application.
