Note: Descriptions are shown in the official language in which they were submitted.
BIS-H~DROXY~ETHYL CARBO~ATE
BRIDGED AMTIBACTERIAL AG~MTS
The invention relates to new chemical compounds
which are o~ value as antibacterial agent~. More
particularly, it relates to novel bis esters of
hydrox~methyl carbonate, HOCH20COOCH20H, in which one
hydroxy group i~ esterified with the carboxy group of
penicillanic acid 1,1 dioxide and the other hydroxy
group i~ esterified with the carboxy group of an
-10 ~ ;ami~openicillin.
Penicillanlc acid 1,1 dioxide ~sulbactam) is
- known from U.S. 4,234,579 to be an effective beta-
lactamase i~hibitor and antibacterial agent.
In UDS~ 4/244,9~1 and BritiSh Patent Application
~oO ~,044,255 bis e~ters of ~he formula
N~ ~ CH3
~ ~ C~3
0~ " COO
(VII)
O O CH
o~~ ~ N ~ ' 3 /
" COO
.
are disclosed in which sulbactam i9 coupled to k~own
a~tibacterial penicillins via methanediol~ In the
above ormula Rl rep~Psants ce~tain ~cyl groups of
~i~
~ .
4~
-- 2 --
known antibacterial penicillins e.y., 2-amino-2-phenylacetyl
or 2-amino-2~ hydrocyphenyl)acetyl.
In copending Canadian application Serial NoO
410,940 filed September 8, 1982 and assigned to the same
assignee, compounds of formula (VII) are disclosed wherein
Rl iS
R2COO- ~ _ CHCO
NH2
and R2 is certain alkyl or alkoxy groups.
In U.S. 3~928,595 antibacterial compounds are
disclosed in which two penicillin or two cephalosporin molecules
are bridged via a carbonate ester. These compounds are of the
formula
R3COOCHOCOO(~HOCOR3
R4 R4
where R~ is H, CE3 or C2H5 and R3 is the residue of a penicillin
or cephalosporin.
Ampicillin, 6-[D- (2 amino-2-phenylacetamido)]-
penicillanic acid is disclosed in U.S. 2,985,648. Amoxicillin,
6-~D-(2~amino-2-[~-hydroxyphenyl~acetamido)~penicillanic acid
is known from U.S. 3,192,198 and U.S. Reissue 28,744. p-Acyl
derivatives o~ amoxicillin are disclosed in U.S. 2,985,648,
U.S. 3,520,876 and U.S. 4,053~360.
~,.
.
~L2~
The present i~vention provides antibacterial
compou~ds o~ ~he formula (I) which are ef~iciently
absorbed rom ~he ga~rointe~inal tract o~ mammals
and aftex absorp~ion are rapidly ~ran~ormed into -the
companent alpha-aminobenzylpenlc~llln ~e..g ampicillin
~r amoxicillin~ and peni~illanic acid 1,l-dioxide
(su~bactam). Said inventio~ compounds are o~ the
~ormula
Rl ~--CHCON~ <C~I3
COOCX2o
~ (I3
O O C30
~ C~3
or a pharmaceutically acceptable acid addition salt
thereof, whexein Rl is hydrogen, hydroxy, formyloxy,
alkanoyloxy having rom two to seven carbon atoms,
alXoxycarbonyloxy having from two to seven carbon
atoms or R2C6H4COO where R2 is hydrogen, alkyl having
from one to ~our caxbon atoms, alXoxy ha~ing from one
to four carb~n atoms, F, Cl, Br, I or CN.
Particularly pre~erred values for Rl include
hydrogen, hydroxy, acetoxy, pivaloylo~y or isabutyryl
ox~r .
L4~
-- 4
The invention also provides valuable intermediates
of the formula (II)
Rl ~ - CHCONU N _ ~ 3 ~II)
COOCH20
O O
\ / C=o
CH3
O COOCH20
wherein Rl is as defined above for compound (I) and Q is a group
readily convertible to amino, preferably azido, benzyloxycarbonyl-
amlno, 4-nitrobenzyloxycarbonylamino or 1-methyl-2-methoxycarbon-
ylvinylamino.
The invention further provides valuable intermediates
of the formula
O /0
1 ~ CH (III)
O COOCH20COOCH2X
wherein X is a good leaving group, perferably Cl, Br or I.
The invention provides a process for production o~ the
compound (I) or a pharmaceutically acceptable acl~ addition salt
thereof, the process being characterized in tha-t:
a. compounds of the -Eormulae
Rl ~ CECONE1~ ~ ~ C}13 ~IX)
N ~CoOZ
and
O O
oF~ooz ( x )
wherein Rl is as defined above, Q is azido, benzyloxycarbonylamino,
4-nitrobenzyloxycarbonylamino or l-methyl-2-methoxycarbonylvinyl-
amino; one of zl and z2 is a carboxylate salt forming cation, M,
and the other is CH20COCH2X where X is Cl, Br or I; are
contacted in the presence of a polar organic solvent to provide
an intermediate of the formula
~ s
3~ .6~
- 5a -
Rl ~ /~ OOCH O (II)
O o
\ / CCO
~ ~ ~ CH3
O COOCH20
and conversion of said intermediate to the product of formula (I)
by conversion of said group Q ~o an amino group, or
b. a compound of the formula
R ~ -CHCONH ~ CH3
o ~/COOCH O
2 \ (VIII)
3 O O
R4 ~ CH ~ =O
N
// ~' COOCH20
wherein Ql is azido, benzyloxycarbonylamino, p~nitrobenzyloxy-
carbonylamino;
~L2~
- 5b -
R is H, Cl, Br o~ I and
R4 is Cl, Br ox I; is contacted with hydrogen in ~he
presence of a hydrogenation catalyst, and
if required, a compound o~ ~ormula ~I) thus~produced
by any process above is converted to a pharmacelltically acceptable
acid addition salt thereof.
The invention also provides a process for production
o the compound (II), the process being characterized ln that:
compounds of the formulae
Rl ~ CHCoNH ~ C33 (IX)
O ~COOZ 1
and
O /0
S / CH3
r COOz 2 (X)
wherein Rl is as de~ined above, Q is azido, benzyloxycarbonylamino,
4 nitrobenzvloxycarbonylamino or 1-methyl-2-methoxycarbonylvinyl-
amino; one of zl and z2 is a carboxylate salt forming cation, M,
and the other is CH20COCH2X where X is Cl, Br or I; are
o
contacted in the presence o~ a polar organic solvent.
~, ~
- 5c -
The invention further provides a process for production
of the compound (I~I), the process beiny charaaterized ln that:
a compound of the formula
O O
\ / CH
F~CH (V
O COOM
is contacted with a compound of the formula XCH2OCOOCH2X, where
M is a carboxylate salt forming cation r in a reaction inert
organic solvent at a temperature of from -20 to 60C.
Compounds of the formula
1 ~ (IV)
R ~ CHCONH ~ ~ CE3
~ COOC~I20COOCH2X
wnere Rl .is as defined above, Q is as defined above for compounds
of formula (II), are also valuable intermediates.
This invention relates to derivatives of penicillanic
acid which is represented by the following structural formula:
H CH3
1--~S~
~ ",/ 3
Q COOH
., ,
~2~
- 5d -
In derivatives of penicillanlc acid, broken line attachement
(''') of a substituent to the hicyclic nucleus indicate~ that ~he
substituent is b~low the plane of the nucleus. Such a sub~tituen-t
i5 said to be in the alpha conEiyuratlon. Conversely, broad line
attachment ( ~ ) of a substituenk to the bicyclic nucleus indic-
ates that the substituent is above the plane of the nucleus. This
latter confiyuration is referred to as the beta-configuration. As
used herein a solid line attachment ( ) of a substituent to the
bicyclic nucleus indicates that the substituent can be in either
the alpha-configuration ox -the beta-confiyuration.
Compounds of the formula (I)-(IV) are named as diesters
of carbonic acid. For example, the compound of formula (I) where
R is hydroyen is desiynated as 6-(2-amino 2-phenylacetamido)pen-
icillanoyloxymethyl l,l-dioxopenicillanoyloxymethyl carbonate;
the compound (II) where Rl is
-6-
hydroxy and Q is azido i5 designated as 6-~2-azido 2-
~-hydroxyphenyl)acetamido3penicillanoylox~me-thyl
l,l-dioxopenicillanoyloxyme~hyl carbonate; and the
compound o~ formula (III) where X is iodo i5 desig~ated
S as iodomethyl l,l-dioxopenicillanoylox~f~ethyl carbo~ate~
Additionally, throughout this ~peci1catlon,
whenever reference .i~ made to compound of formula (I),
(II) or (IV), if not already 90 indicated, it i~
understood ~hat ~his reers to a compound in which
the substituent
Rl- ~ OElCON~ ,
wherein Q2 is amino or Q and Rl and ~ are as defined
above, has the D-configuration.
The compounds of fQrmU 1 a (I) can be prepared by
many of the methods known in the art for synthesis o
ester~. ~owever, the preferred general method
involves salt formation by condensation of a carboxy-
late sal~ with a halomethyl e~ter~ Two preferred
~uch met~ods are outlined below.
. ~
--7--
~--C~CO~ ~
'COOM ~T 'COOCH20
(VI) (III) C--O
~CCH ~O
Rl -~-CHCO~H,~ ~}3
N 'COOCH2o
O O
S C}~3
O'~ COOC~
( I I ~
Re~nove protecting
group from Q
(I)
E~ C~cO~C~i3 ~ K3
C:Os: C~2o~ o N 'COQM
,,5~=
~)~{C~;~o tV)
. r
: ~ ~I ) ~(I)
-8-
In the above ormulae, Rl and Q are as
defined above, M i5 a carboxylate salt forming
cation, preferably Wa, K or ~(C4Hg~ ca~ions and X i9
a go~d leaving group, preexably C1, Br or I.
The group Rl in the above intermediates o~
formulae (IV) and (VI) and products (II) and (I)
includes those compounds wherein RL is acyloxy and
alko~ycarbonyloxy as de~ined above. The carboxylic
acid precursors of such intermediates (VI) can be
prepared e.g., by methods des~ribed in U.S. 4,053,360
by acylation of 6-aminopenicillanic acid with the
appropriate acid of the fonmula
Rl- ~ -C~COOH
where Rl and Q are as define~ ab~ve, or an activated
derivative thereof, e~g. the acid chloride or mixed
anhydride ~ormed with ethyl chloroformate. In the
res~lti~g i~vention compounds ~I) and'(II) obtàined
by the abo~e reactions, Rl ha.~ the same value as the
star~in~ material of fonmula tVI).
~ Alternativelyt the starting material of formula
(IV) or (VI~ can b~ one in which Rl is hydroxy and
the resulting intermediate o formu}a ~II) i~ sub-
se~uently acylated or`alkoxycarbonylated to fo~m the
correspondi~g compound of formula (II) wherein Rl is
alkyl~arbonyloxy, alkoxycarbonyloxy or R2C6-H4COQ as
defi~ed above.
- 9 -
The acylation or alkoxycar~onylation o the
intexmediate of ormula (II) w~erein R1 is hydroxy
and Q is as previously defi~ed can be carried ouk
e.g., by reac-~ing said compound o~ ~ormula (II) with
,the appropriate acid chloride or acid anhydride. The
reaction is ordinarily carried out in the presence o~
a reacti.on-inert solvent sys~em. In a typical
procedure, from 0~5 to 2.0 molar equivalents, and
preferably about 1 molar equivalent, ~f the appro-
priate acid chloride or acid anhydride is contactedwith the starting compound of formula ~ wherein
i~ hydro~y, in a reaction-inert solvent, in the
presence of a ter~iary amine, at a temperature in the
~ange from -10 to 30C~ Reaction inert solvents
which can be used in this acylation are chlorinated
hydrocarbons, such as chloroform and dichloromethane;
eth~rs, such as die~hyl e~h~r and tetrahydrofuran;
}ow molecular weight esters, such as ethyl acetate
a~d butyl acetate; low moleculax weight aliphatic
ketones r such as acetone and methyl ethyl ketone:
~ertiary amides, such as ~,N-dimethylormamide a~d
methylpyrrolidone, acetonitrile; and mixtur~s thereof.
The tertiary ~mine is normally used in an amount
equivalent to the starting acid chloride or acid
anhydride, and typical tertiary amines which can be
used axe triethylamine, tributylamine, diisopropyl-
ethylamine~ pyridine and 4-dime~hylaminopyridine~
10--
In each of the reaction sequences designated as
A and B, above, to form the amino-protected products
o~ formula (II), the respective carboxylate salt and,
e.g., halomethyl ester are contacted in approximately
equimolar amoun~s in ~he presence of a polar organic
solvent at a temperature of from about 0 to 80C.
and preerably from 25 to 50C. While at leas~
equimolar amounts of reactants are ordinarily employed,
an excess of the carboxylate salt, up to a ten-fold
molar exces~, is preferred. A wide variety of solvents
can be used for this reaction, however, it is usually
advantageous to use a relatively polar organic
solvent to minLmize the reaction time. Typical
solve~ts which can be employed include ~ dimethyl-
formamide, N,N-dimethylace~amide, ~-methylpyrrolidone,
dimet~ylsulfoxide, ethyl acetate, dichlorom~thane,
chloroform, acetone and hexamethylphosphoric triæmide.
~he time required for the reactio~ to reach substan-
~ial completio~ varies according to a number of
factors, such as ~he ~lixture of the reac~ants, the
reaction temperature and solvent. However, at about
25C~ reaction time~ of from about 10 minutes to
about 24 hours are commonly emploved.
The desired amino-protected compound of formula
(II3 is then isolated by methods well known to those
~ skill in the art. For axample, the reaction
mi~ture is taken up in a water Lmmi3cible solvent,
e~. ethyl acetate, chloroform or methylene diohloxide,
washed with water, brine and dried. Evaporation of
solvent provide~ the int2nmediate o formula ~
which can be purified, if desired, e~g. by chromato-
graphy o~ silica gel.
:: :
~2~4~
The removal of the amino-protecting group from
the intermediate (II) is carried out by methods well
known in the art, see, e.g. Gross e-t al. in "The
Peptides, A~alysis, Synthesis, Biology", Academic
Press, ~ew York, N.Y., Vol. 3, 1981, but due regard
mu~t be given to the lability of the beta-lactam ring
and the ester linkages. For example, when Q is 1-
methyl-2-methoxycar~onylvinylamino, the protecting
group ~l-methyl-2-~ethoxycarbonylvinyl) can be
removed ~Lmply by treating the compound of formula tII)
with one equivalent of a strong aqueous acid, e.g.
hydrochloric acid, in a re~ction inert solvent, at a
temperature in the range o~ from -10 to 30C. In a
typical procedure, the compound of formula ~ is
treated with one equivalent of hydrochloric acid in
aqueous acetone. The reaction is usual}y complete
within a short ~Lme, e.g~ within one hour. Then the
acetone is removed by evapora-tion in vacuo, and the
me~hylace~oacetate by-product is removed by extraction
with ether. Finally, the compound of ormula (I) is
recovered by lyophilization as its hydrochloride
salt.
Compounds o ormula (II) wherein Q is a~ido,
ben~ylox~carbonylamino or 4-nitrobenzyloxycarbonyl-
amino can be converted to the corresponding minocompound of for~ula (I) by subjecting the compound
~II) to conditions commonly ~mployed for catalytic
hydrogenolysis. The compound o~ formula (II) is
stirred or shaken ~nder an atmosphere of hydrogen, or
hydrogen, optionally mixed with an inert diiuent such
a~ nitrogen or argon, in ~he presence of a catalytic
æmount o~ a hydrogenolysis catalyst. Convenient
solvents for this hyd.roge~olysis are lower-alkanols t
su~h as methanol and isopropanol; ethers, such as
~2~
-12-
tetrahydrofuran and dioxan; low molecular weight
e3ters, such as ethyl acetate and butyl acetate;
chlorinated hydrocarbons, such as dichloromethane and
chloroform; water; and mixtures of these soLvents.
S However, it is usual to choose conditions under which
the starting material is soluble. The hydrogenolysis
is usuaLly carxied out at a temperature in the range
from 0 to 60C. and at a pressure in the range from 1
to 10 atmospheres, preferably about 3 4 at~ospheres~
The catalysts used in this hydrogenolysis reaction
are the type of agents known in ~he art ~or this kind
of transformation, and typical examples are the noble
metals, such as nickel, palladium, platinum and
rhodium~ The catalyst is usually used in an amount
from 0.5 to 5.0, and preferably about 1.O, times the
weight of the compound of formula (II). It is often
convenient to suspend the catalyst on an inert
suppor~; a particularly convenient cataly~t is
palladium suspended on an inert support such as
carbon.
The compounds of the ~ormula (I) will form acid
addition salts, and these acid addition salts are
con~idered ~o be within the scope and purview of this
invention. Said acid addition salts are prepared by
stan~ard methods for penicillin compounds, for
example by combining a solution of the compound of
formula tI) in a suitable solvent (e~g. watar, ethyl
- acetate, acetone, methanol, ethanol or b7~tanol~ with
a solution containing a stoichiometric equivalent of
the appropriate acid. If the sal~ precipitates, it
is recovered by filtration. Alternatively, it can be
recovered by evaporation o~ the solvent, or, in the
case of aqueou~ solutions, by lyophilization. Of
particular value are ~he sulfate, hydrochloride,
hydrobromide, nitrate, phosphate, citrate, tartxate,
- pamoate, perchlorate, sulfosalicylate, benzenesulfona~e,
4-toluenesulfonate and 2-naphthalenesulfonate salts~
The compounds of the formula (I), and the salts
thereof, can be purified by conventional methods or
penicillin compounds, eOg. recrystallization or
chromatography, but due regard must be given to the
lability of the beta-lactam ring systems and the
ester linkages.
An alternate process for preparation of the
antibac~erial compounds of formula (I) employs an
intermediate of formula
~ Ql r~ CH3
N 'COOC~20
R3 ~/ C~ C=O ~ VIII)
R ~ C~ ~
where Rl is as previously defined, Ql is azido,
benæyloxycarbonylamino or E~ni~robenzyloxycarbonyl-
amino, R3 is H, Cl, Br or I and R4 is Cl, Br or I.
The intermediate (~III) upon catalytic hydrogenation,.
e.g. by ~he me~hod described above for hydrogenolysis
of azido, or ben~yloxycarbonylamino compounds of
formula (II), is simul~aneously hydrogeno~yzed at the
Q , R~ and~or R4 substituents to provide the inventio~
compound o formula (I).
~Z~4~6~L
-14-
The intermediates (VIII) are obtained by methods
analogous to t~ose described herein for preparation
of intermediates of ormula (II), but employing a
R3,R4-substituted l,l-dioxopenicillanate in place of
the corresponding unsubstituted l,l-dioxopenicillanic
acid, its salts or derivatives of formulae (III) or
(V) .
Methods for preparation o~ the requisite R3,R4-
disubstitu~ed l,l-dioxopenicillanic acids and salts
thereo are taught in U.S. 4,234,579; British Patent
Application 2,044,255 and Belgian Patent No. 882,028.
The intermediates o~ formula (I~) are obtained,
for example, as outlined below.
OCH X
OCH2X -C~CO~H ~ ~ CH3
. o 'COOM
-L~X ~VI)
~ ` ~ r
R ~ -C~CO~H ~ < C~3
O '~COOCH20COOCH2X
(I~)
where Rl t M, Q and X are as defined above. The
reaction is carried out by contacting the starting
amino-protected benzylp~nicillin satt of formula (VI)
with a~ least an equimolar amount, pre~erably a molar
-15-
e~cess of up to te~-fold, of a bis-halomethyl carbonate
in the presence of a reaction inert organic solvent
and a temperature of from a~out -20 to 60C., prefer-
ably from 0 to 30C. The ~olvents w~ich can be
employed succ~ssfully in this reaction include the
~ame polar organic solvents employed above for
prPp~ration of interm~diates of formula (II).
The intermediate halomethyl l,l-dioxopenicillan-
oyloxymethyl carbonate~ of formula (III) are prepared
as described above for the intermediates of formula (IV~,
but employing a salt of penicillanic acid 1,l-dioxide
of formula ~V) as starting material i~ place o~ the
s~arting penicillin salt of ormula (VI).
bi~-Chloromathyl ~arbonate is prepared by photo-
chemical chlorination of dimethyl carbonate by the
method of Rling _ al., Compt. rend., 170, 111, 234(1920), ChemO Abstr., 14, 1304 (1920). bis-Bromo~
methyl carbonate and _ -iodomethylcarbonate are
o~tained from the bis-chloromethyl compound by
~ reaction with e~g. sodium bromide or potasssium
iodi~e by methods well known in the art.
The carboxylate salts of ormulae t~) and (VI)
are obtained from the corresponding carboxyli~ acids.
A pra~erred method ~or providing the salts wherein
is an alkali metal such as sodium or potassium
employs the appropriate salt of ~-ethylhexanoic acid
a~ base. Typically, the carbo~ylic acid of formula (V)
or (VI~ is dissolv~d in ethyl acetate, a~ equimolar
amount o~ ~dium or potassium 2-ethylhe~anoate is
3~ added with stirring and the pr~cipitated salt of
formula (V) or (VI) collected by filtration.
~2~
-16-
The corresponding sal~s of formula (V) or (VI)
where M is tetrabutylammonium can be obtained rom
the corresponding acid, e.g. by neutralization with
aqueous tetrabutylammonium hydroxide in the presence
of a water immiscible organic solvent, preferably
chloroform. The solvent layer is separated and the
product isolated by evapora~ion of solvent. Alternately,
~he sodium or potassium salts o~ formula (~) or (VI)
are reacted with an equimolar amount of aqueous
tetrabutylammonium hydrogen sulfate in the presence
of a water immiscible solvent, the preciptiated
al~ali metal bisulfate salt removed by filtration and
the product isolated by evaporation of solvent.
While in the present i~vention the antibacterial
compounds of ~he formula (I), as defined above, are
the preferxed compounds wherein a penicillin and a
beta-lactamase inhibitor are linked as a bis ester of
hydroxymethyl carbonate, in a broader sense th~
invention relates to antibacterial compounds of the
general formula
Rl ~ ~CHCO c~3
NH2 ~ ~ CH3 RS
0~ 'COOCHO
\ -- (IX)
C=O
B-COOCHo
RS
where RS is hydrogen or alkyl having from one to
three carbon atoms, B is the residue of cartain beta-
lacta~ase inhibitors and Rl is as defined above.
6~
Examples of beta-lactamase inhibitors, B,
include
CH3 , ~
(a) (~)
O O
\/ EI
F~CH2Y X ~< 3
where ~ is Cl or CH3COO where Xl = CL, Br or I
(c) (d)
and
O_ " ~CH 2OH
~ r
0~ 1 "
(e)
-18-
The preferred compounds of formula (I), those
above wherein the beta-lactamase inhibitor residue,
B, is l,l-dio~openicillanoyl, (a), are prepared, as
de~cribed above, employing hydrogenolysis me~lods.
Analogous compounds of ormula (IX~ wherein B is.
moiety (b), above are sLmilarly obtained. However,
certain of the above moieties B are no~ stable to
hydrogenolysis. Thus, preparation of compounds of
formula (IX) w~erein B is a moiety that is unstable
to hydrogenolysis conditions such as tc), ~d) or (e),
above, requires the use of a protecting group which
is removable by mild hydrolysis, or example, as
outlined below.
-
R~ CHCO~ S CH3
/ ' 6 ~ ~ ~'
'==J NHR ~ ~ CH3 + BC02M
.. . . . . ' C02C~120COOCH2~
(X~ -MX '
.
~ ~ '
R~ C~CONH~ ~ C~3 H O
F ~1 ~ " 3 ~ (IX)
C02CH 20~
~0
(XI)
BCC)OCH20
where B, M, Rl and X are as d~ined above and R6 is
an ~m~no protecting group whi~h is removable by mild
hydrolysis, e.g., triph~nylmethyl or an enamino group
such as
--19~
CH3~¢
oR7
where R7 is alkoxy having from onë to three carbon
atoms or aminoO
Alternately, of course, the compounds of formula (IX~
S which are not stable to hy~rogenolysis conditions can
be prepared by the following methods.
BCOOC}~20COC~2X + R --~--CHCO~E~ =C~o33M
-MX
~ r
(XI)~ (IX)
where B, M, ~1, X and R6 are as de~ined above.
When contemplating therapeutic use for a salt of
lQ an antibacterial compound o~ ~his invention, it is
necessary to use a pharmaceutically-acceptable salt;
however, salts o~her than these can be used ~or a
variety o~ purposes. Such purposes include isolating
and purifying particular compounds, and interconv~rting
phanmaceutically-acceptable salts and their non-~alt
counterparts.
~2ï~
-20-
The compounds of formula (I) and pharmaceutically
acceptable acid additi~n salts thereo~ possess ln
v _ antibacterial activity in mammals, and this
activity can ~e demonstrated by standard techniques
S for penicillin compounds. For example, the compound
of formula (I3 is administered to mice in which acute
infections have been established by intraperitoneal
inoculation with a standardized culture of a pathogenic
bacterium. Infection severity is standardized such
that the mice receive one to ten times the LDloo
tLDloo: the minimum inoculation req~ired to consistent-
ly Xill 100 percent o~ con~rol mic~)~ At the end of
the test, the activity of the compound is assessed by
counting the number of survivors which have been
challenged by the bacterium and also have received
the i~vention compound. The compounds of formula (I~
can be administered by both the oral (p.o.) and sub-
cutaneous (s.c.~ route.
The in vivo activity of the antibacterial
~0 compounds of this invention makes them suitable for
the control of bacterial in~ections in mammals,
including man, by bo~h the oral and parenteral mcdes
o~ administration.. The compounds are usaful in the
control of infections caused by susceptib}e bacteria
in human subjects.
~2 ~
-21~
A compound of formula (I~ wherein Rl is other
than hydrogen brea~s down to 6-(2-amino-2-~4-hydroxy-
phenyl]acetamido)penicillanic acid (amoxicillin~ and
penicillanic acid l,l-dioxide (sulbactam) after
administration to a mammalian subject by both the
oral and parenteral route. Sulbac~am then functions
as a beta-lac~amase inhibitor, and it increases the
anti~acterial effectiveness of the amoxicillin.
Similarly, a compound or formula (I) wharein Rl is
hydrogen breaXs down to 6-(2-amino-2-phenylacetamido)-
penicillanic acid (ampi~illin) and sulbactam. Thu5,
the compounds of the formula (I) will find use in ~he
control o~ bacteria which are s~?sceptible to a 1:1
mixture of amoxicillin and sulbactam or ampicillin
and sulbactam, e.g. susceptible strains of Escherichia
coli and Staphylococcus aureus.
. =
In determining whether a particular strain of
Escherichia coli or Staphylococcus aureus is sen
,
sitive to a particular therapeutic compound of the
invention, the in vivo ~est described earlier can be
used, Alternatively, e~g., ~he minimum inhibitory
concentration (MIC) o a 1:1 mi~ture of amo~lcillin
and sulbactam or ampicillin~sulbactam can be measured.
l~e MIC's can be measured by the procedure recommended
by the Interna~ional Collaborative Study on Antibiotic
Sensitivity Testing ~Eri~cson and Sherris, Acta.
et Microbiolo~ia Scandinavica, Supp. 217,
_ . _
Section B: 64-68 ~1~71]), which employs brain heart
infu~ion (B~I) agar and the inocula replicating
device. Overnight growth tubes are diluted 100 rold
for use as the standard inoculum (20,000-10,000 cells
in appro~imately Q.002 ml. are placed on the agar
surfaca; 20 ml. of B~I agar/dish~. Twelve 2 fold
~2~
-22-
dilutions o~ the test compound are employed, with
i~itial concentration of the t~st drug being 200
mcg./ml Single colonies are disregarded when
xeading plates after 18 hrs. at 37C. The suscept-
ibility (MIC) of the test organism is accepted as thelowest concentration of compound capable of producing
complete inhibitlon of growth as judged by the naked
eye.
When using an antibacterial compound of this
invention, or a salt thereof, in a mammal, parti-
cularly manr the compound can be administered alone,
or it can be mixed with other antibiotic substances
a~d/or pharmaceuticalLy-acceptable carriers or
diluents~ Said carrier or diluent is chosen on the
basis of the intended mode of administration. For
example, when considering the oral mode of administra-
tion, an antibacterial compound o this invention ~an
be used in the form of tablets, capsules, lozenges,
troches, powders, syrup3, eli~irs, aqueous solutions
and suspensions, and the like, in accordance w~th
standard pharmaceutical practice. The proportional
ratio of active ingredient to carrier will naturally
depend on the chemical nature, solubility and stability
o the ac~ive ingredient, as well as the dosage
con~emplated. In the case o tablets for oral use,
carriers which are commonly used includa lactose,
sodium citrate and salts of phosp~oric acid. Various
di integrants such as s~arch, and lubricating agents,
~uch as magnesium stearate, sodium lauryl sul~ate and
talc, are commonly used in tablets. For oral adminis-
tration in capsule orm, useful diluents are lactose
and high molecular weight polyethylene glycols, e.g.
polyethylene glycols having molecular weights of from
~.2~6~
-23-
200Q to 4000. When aqueous suspensions are requirea
for oral use, the active ingredient is combined with
emulsifying and suspending agents. If desired,
certain sweetening and/or flavoring agents can be
add~d. For parenteral administration, which includes
intramuscular, intraperitoneal, subcutaneous, and
in~ravenous use, sterile solutions of the active
ingredient are usually prepared, and the pH of the
~olutions are suitably adjusted and buffered. For
intravenous use, the total concentration of solutes
should be controlled to render the preparation
isotonic.
A~ indicated earlier, the antibacterial com-
pounds of this invention are of use in human subjects
and the daily dosages to be used will not differ
significantly from other~ clinically-used, penicillin
antibiotics. The prescribing physician will ultimately
determine the appropriate dose for a given human
subject, and this can be expected to vary according
to the age, weight, and response of the individual
patient as well as the nature and the severity of the
patient's symptoms. The compounds o this invention
will normally be used o~aLly at dosages in the range
rom 20 to about 100 mg. per kilogram of body weight
per day, and parenterally at dosages rom about 10 to
about 100 mg. per Xilogram of body weight per day,
usually in divided dQses. In some instances it may
be necessary to use doses outside these ranges.
~2~
-24-
The following examples and preparations are
provided solely for further illustration. Nuclear
magnetic resonance spectra (NMR) were measured for
solutions in deuterated chloroform (CDC13) or deuterated
dLmet~yl sulfoxide (DMSO-d6), and peak positions are
reported in parts per million downfield from tetra-
methylsilaneO The following abbreviations for peak
shapes are used: bs, broad singlet; s, singlet; d,
doublet; t, triplet; q, quartet, m, multiplet~
6~
-~5
EXAM~LE 1
bis-Iodomethyl Carbonate
To a solution of 10.7 ml. (15.9 g., 0.1 mole~
bis(chloromethyl)carbonate in 400 ml. acetone was
added 75 g. (0~5 mole) sodium iodide. The mixture was
re~lu~ed for 2 hours under nitrogen, then lert over-
~ight at room temperature. The mi~ture was filtered
a~d the fil~rate was concentrated ln vacuo. Methylene
chloride ~500 ml.) was added and the resulting mixture
was filtered. The filtrate was concentrated to about
200 ml., 200 ml. of water was added and the aqueous
phase was adjusted to p~ 7.5. Aqueous sodium thio-
sulfate solution was added to rPmova iodine, the
organi~ phase was separated and dried over sodium
sulfate. The dried methyLene chloride solution was
concentrated _ vacuo to an oil which darkened on
standing. The oily product was treated with a mixture
of 35 ml. he~ane and 6 ml. diethyl ether at 0C., the
resulting crystals war~ filtered, washed with hexane
and dried, to afford 10.0 g. ~29%) of yellowish
crystalline product. M.P. 49-51C. lH-NM~ (CDC13~
ppm (delta): ~.94 singlet; infrared spectrum (~ujol)
cm 1 1756, 1775~
~2~
-26-
EXAMPLE 2
Iodomethyl 6-~D-(2-azido~2-phenyl-
acetamido)~penicillanoyloxymethyl carbonate
To a cooled solution (0C.) of 2.43 g. (7.1
mmoles) of bis-iodomethyl carbonate in chloroform
~16 ml~), a solution of 2.19 g. (3.5 mmole) of tetra-
butylammonium ~_CD_( 2-azido-2-ph~nylacetamido)penicil-
lanate in 10 ml. chloroform was added dropwise. After
the addition was completed the reaction mixture was
allowed to warm to room temperature and allowed to
stand at room temperature overnight. The solvent was
removed in vacuo and the residue was chromatographed
on silica gel, eluting with 8:1 by volume me~hylene
chloride/ethyl acetate to yield 822 mg. (40~) of
product. lH-~MR tCDC13) ppm (delta): 1.52 (s, 3~),
1.65 (s, 3~), 4.45 (s, lH), 5.04 ~s, 1~), 5.65 (m,
4~), 5 . 92 ( s , 2H), 7.34 (s, 5H); infrared spectrum
- (C~C13): 1770 ~m 1,
~2~
EXAMPLE 3
6-~D (2-azido-2-phenylacetamido)~-
penicillanoyloxymethyl 1,L-
dioxopenicillanoyloxymethyl Carbonate
~II, Rl = H, Q = ~ )
_ -- . 3
. To a solution of 822 mg. (1.4 mmole) iodomethyl
6-CD-(2-azido-2-phenylacetamido)]penicillanoyloxymethyl
carbonate in 30 ml. chloroform was added dropwise at
room temperature a s~lution of 1.33 g. (2.8 mmole)
tetrabutylammonium 1,1-dio~openicillanate in 30 ml.
chloroformO The raa~tion mixture wa~ stirred at room
temperature overnight, concentrated in vacuo and
chromatographed on silica gel. Elution with 9:1 by
volume methylene chloride/ethyl acetate gave 0.51 g.
o~ product (52.6% yield). 1H~MR (CDC13) ppm (d~lta):
1.4 (s~ 3EI) I 1.5 (s~ 3H~ r 1.6 (Sr 3EI) ~ 1~64 (s~ 3E~) ~
3.42 (d, J = 3~z, 2~), 4~4 ~s, lH), 4.44 (s, lH), 4.6
~t, J ~ 3~zl lH), 5.04 (s, 1~), 5.44-6.0 (m, 6~) r 7.35
(s, 5~), infrared spectrum (CHC13): 1775 cm 1.
-2~-
-EXAMPLE 3A
When the procedure of Example 2 is carried out
with the tetrabutylammonium salt o 6-~D-(2-benzyl-
oxycarbonylamino-2-phenylacetamido)~penicillanate in
place of the corresponding azidocillin salt, the
product obtained is iodomethyl 6-[D-~2-benzyloxycar-
bonylamino-2-phenylacetamido)~penicillanoyloxymethyl
carbonate. When the latter compound is employed as
starting material in the procedure o~ Example 3, the
product obtained i 6-~D-(2-benzyloxycarbonylamino-2-
phenylacetamido)~penicillanoyloxymethyl 1,l-dioxo-
penicillanoylox~methyl carbonate.
Similarly, starting wtih the 6-CD-(2-~-nitro-
benzyloxycarbonylamino-2-phenylacetamido)~penicillanate
LS6 salt in ~he method of Example 2 and carrying the
product through the method of Example 3, provides 6-
~D-(2-~-nitroben~yloxycarbonylamino-2-phenylacetamido)~-
penicillanoyloxymethyl L,l-dioxopenicillanoyloxymethyl
carbonate.
-29-
EX~M2LE 4
. . ~.
6-CD-(2-amino~2-phenylacetamido)]~
penicillanoyloxymethyl 1,1-dioxo-
penicillanoyloxymethyl carbonate
t I r Rl = El )
A -~olution of 1.49 gO 6~-~D-(2-azido-2-phenyl-
acetamido)~penicillanoylox~methyl l,l-dioxopenicillan-
oyloxymethyl carbonate in 40 ml. methylene chloride
and 20 ml. isopropanol was hydrogenated at 60 psi
(4.2 kg./cm.2) in ~he presence of 1.5 g. 10% Pd/C for
30 minutes. A fur~her 1.5 g. of catalyst was added
and the hydrogenation ~as continued or another
30 minutes. The catalyst was then filtered off and
the filtrate was concentrated to provide a white solid
residueO The residue was dissolved in tetrahydrofuran/
water (1:1), the resulting solu~ion was cooled to 0C.
and adjusted to p~ 2.5 with 0.1~ hydrochloric acid.
The t~trahydrofura~ was- evaporated i~ vacuo and the
resulting aqueous solution was freeze dried to afford
680 mg. (45~) of proauct as a white solld. l~-NMR
(CDC13 + CD30D) ppm ~delta): 1.42 (s, 6H), l.S (s,
3~), 1.6 (g, 3H), 3.46 ~m, 2~), 4.4 (s, 1~), 4.43 (s,
lH), 4.72 (m, lH3, 5.2 (s, lH), 5.48 (q, ~ = 4Hz, 2H),
5.82 (~ + q, J = 6~z, 4~), 7.44 (s, 5~), inrared
25 . spectrum (~ujol): 1775 cm 1.
When ~he ~orresponding benzyloxycarbonylami~o
c~mpounds provided in Example 3A are employed ~s
starting material in the abo~e procedure, the same
title compound i9 obtained in like manner.
-30-
EXAMPLE 5
Chlorome-thyl 1,l-dioxo-
penioillanoyloxymethyl carbonate
To a mixture of 1.17 g. t5 mmole) penicilLanic
acid l,l-dioxidet 50 ml. chloroform and 10 ml. water
is added 40~ aqueous tetrabutylammonium hydroxide with
vigorous ~irring until a p~ of 8.5 is obtained. The
chloro~orm layer is separated and the aqueous phase
extracted with fresh chloroform. The combined organic
layers are dried and coi~centrated to a small volume
(about 20 ml~).
To a solution of 1.5 g. (10 mmole) bis-chloro
methyl carbonate in 15 ml~ chloroform at 0~. is added
dropwise the above solution of tetra~utylammonium 1,1-
dioxopenicillanate. Ater the addition is completed,the mixture is allowed to warm to room temperature and
stir overnight. The chloroform is evaporated in acuo
and the cruae product puriied by chromatography on
5 ilica gel~
2~
-31-
EXAMPLE 6
Iodomethyl l,l-dioxo-
~enicillanoyloxymethyl carbonate
To a solution of 3.37 g~ (10 mmole) chloromethyl
S l,l-dioxopenicillanoxymethyl carbonate in SO ml.
acetone is added 7.5 g. (50 mmole) sodium iodide and
the mixture is stirred overnight at room temperature.
The acetone is evaporated in vacuo, the residue
partitioned between water and ethyl acetate. The
aqueous phase is separated, the organic phase washed
with water, brine, dried, (~a2So4) and concentrated in
vacuo to give the iodomethyl compound which is purified,
if desired, by chromatography on silica gel.
When the above procedure is repeated but using
dimeth~Lformamide as solvent in pla~e o acetone and
sodium bromide in place of sodium iodide, bromomethyl
l,l~dioxopenicillanoylox~methyl carbonatP is obtained.
-32-
EXAMPLE 7
6-[D~ methyl-2-methoxycarbonyl-
vinylamino]-2-~p-hydroxyphenyl]-
acetamido)]penicillanoyloxymethyl
5- l,l-dloxopenicillanoyloxymethylcarbonate
(II, Rl = OH, Q = ~HCH(CH3? = C~C2CH3]
A. To 300 ml. of dichloromethane is added 41.9 g. of
6-t2-amino-2-~ A -hydroxyoheny~acet~lido)penicillanic
acid trihydrate and 50 ml. of water, and then the pH
is adjusted to 805 using 40% aqueous tetrabutylammonium
hydroxide7 Three layers are obtained. The upper
layer is removed, saturated with sodium sul~ate and
then it is eætracted with dichloromethane. The
extracts ~re combined with the middle layer and the
lower layer, and the resul~ing mixture is evaporated
in vaouo to give an oil which crystalli7ed on tritura-
tio~ with aoetone. This afforded 44.6 g. of tetra-
butylammonium 6 (~-amino-2-C4-hydroxyphenyl~acetamido)-
penicillanate_
The a~ove salt is added to 150 ml. o methyl
acetoacetate and ~he suspen~ion is heated at ca. 6~C.
un~il a clear solution is obtained ~8 minutes). The
mix~ure is allowed to cool, and the solid recov~red ~y
~iltra~ion. The solid is washed with me~hyl aceto-
acetate, followed by die~hyl ether, to give 49.25 g.
o~ tetrabutylammonium 6-(2~ m~thyl-2-methoxycarbonyl-
~i~ylamino~-2-~4-hydroxyphenyl~a~etamido)penicillanate.
.
B. A mixture of 7.04 g. (0.01 mole) o the tetra-
butylammonium salt o amoxicillin enamine obtained in
Part A, 4.28 g. (0.01 mole) iodomethyl l,l-dioxo-
penicillanoyloxymethyl car~onate and 65 ml. chloroform
is stirred at room temperature for eight hours. The
mixture is diluted with 500 ml. ethyl acetate, washed
wi~h brine, ~er, brine again and dried (~a2S04).
The solvent is evaporated in vacuo, the re~idual
product is dissolved in a minimal amount of ethyl
a~etate and purified by chromatography on silica gel.
EXAMPLE 8
6-CD- (2-~mino 2-~p-hydroxyphenyl~-
acetamido)3penicillanoyloxymethyl
l,l-dioxopenicillanoyloxymethyl
15. . carbonate hydrochloride
(I, Rl ~ ~)
To 2.5 g. 6-{D-(2~ methyl-2-methoxycarbonyl-
vinylamino~-2-Cp-hydroxyphenyL~acetamido)]penicillan-
oyloxymethyL l,l-dioxopenicillanoyloxymethyl carbonate
dissolved in 60 ml. acetone is added 33 ml. 0.1~
hydrochloric acid and the mixture is stlrred or
20 minutes at xo~m temperature. ~he aceto~e is
evaporated in vacuo, the aqueous residue ex~racted
._
with ethyl ether, then with ethyl acetate. The
aqueous layer is then free2e dried to aford the title
hydrochloride salt~
-34-
EX~LE 9
6-~D~(2-Amino-2-phenylacetamido)~-
penicillanoyloxymethyl 1,1-
dioxopenicillanoyloxymethyl
carbonate hydrochloride
(I, R1 = H~
~ , , .
To 300 ml~ chloroorm is added 40.3 g. ampicillin
trihydrate and 50 ml. water~ The p~ is adjusted to
8~5 with 40~ aqueous tetrabutylammonium hydroxide, and
the tetrabutylammonium salt isolated and reacted with
methyl acetoacetate by the method of Example 7, Part A
but using chloroform as solvent in place of dichloro-
methaneO The resulting tetrabutylammonium 6`CD-t~-cl-
me~hyl-2-methoxycarbonylvinylamino~-2-phenylacetamido)]-
}5 penicillanats (obtained in ~2~ yield) is reacted with
iodomethyl l,l-dioxopenicillanoyloxymethyl carbon~te
by the method o Example 7, Part B to provide 6-~D-~2-
~l-methyl-2-mPthoxycarbonylvinylamino~-2-phenylacet-
amido)~penicillanoyloxymethyl 1,1-dioxopenicillanoyl-
oxymethyl carbonate. Removal of the enamine protectinggroup with aqueous hydrochloric acid in acetone by the
method of Example 8 provides the title compound.
The compounds of the formula below are ob-tained
in liXe manner~
--3S--
CECO~H ~;~ CH3
~H2 ,~L--!~ c~3
' COC)CH20
0~0 C=O
o~T ~ COOCEI20/
where Rl is:
CF~3C
C2~5C~
( CE~3 ) 2~HCOQ
CH3 ( CH:2 ) 3C
C~I3 ( C~ 5 COO
CH3 OC00
( C~3 ) 2CH~
( C}~3 ~ 3COC00
C~I3 ( ~2 ) 50CC)0
C6HSCOO
4-FC6H4CoO
2-ClC~H4C00
3-BrC6H4C00
4-C~C6~4Co
3C6H4CO
3 ( C~I3 ~ 2CHC6Hd,COO
4~ I3 ) 3CC6~4C
3-C~I3~6~'1`C~
4-C2~50~*~0
_ 3H70C6H~Ct:)O
3 ( ~13 ) 2CEICH20C~;H4 0
4-n-C4H90C6H4cOO
~ Q0
-36-
EXAMPLE 10
6-~D-(2-Amino-2~ acetoxyphenyl]-
acatamido)]penicillanoyloxymethyl 1,l-dioxo-
penicillanoyloxymethyl carbonate hydroc~loride
(I, Rl = CH COO)
_ 3
A. 6-CD-(2~ methyl-2-methoxycarbQnylvinylamino~-
2-~ -aceto~yphenyl~acetamido)~penicillanoyloxymethyl
l,l-dioxopenicillanoyloxymethyl carbonate_ _
6-~D-(2~ methyl-2-mPtho2ycarbonylvinylamino]-2-
~p-hydroxyphenyl~acetamido)~penicillanoyloxy~ethyl
1,l~dioxopenicillanoyloxymethyl carbonate 2.34 g.
(3 mmole), prepared by the method o ~xample 7, and
0.366 g. t3 mmole) 4-dimethylaminopyridine are dissolved
in 30 ml. dichloromethane and 0.28 ml.. (3 mmole)
acetic anhydride is added. .The solution is stirred
for 30 minutes, diluted to 100 ml~ with dichloromethane,
wa~hed with water and brine, then dried (~a~So4)..
~vaporation of solvent _ vacuo affords the title
ompound..
B. To 1.9 g of the product obtained in Part A,
above, dissolved in 50 ml. acetone is added 23 ml.
0..1~ hydrochloric acid. The resulting mixture is
stirred for 25 minut~s at room temperature and the
acetone evaporated in vacuoO The aqueous phase is
wa~hed with ethyl ether, clariied by ~iltration and
freeze dried to yield the title compound.
-37-
EXAMPLE 11
6-C~-(2-Amino-2-~p-pivaloyloxy-
phenylJacetamido)~-
penicillanoyLoxymethyl 1,1-
dioxopenicillanoyloxymethyl
carbona~e hydrochloride
~I, Rl - (CH3)3CCOO~
The title compound is obtained by repeating the
procedure of Example lO, but using 0.33 g. (3 mmole)
o~ pivaloyl chloride in place of the acetic anhydride
in Part A. The enamine protecting group is removed
from the resulting p-pivaloy}oxyphenyl ester with
aqueous ~ydrochloric acid in acetone and the product
i~olated as des~ribed in Part B of Example lO.
Use of isobutyryl chloride or isobutyric anhydride
n the above procedure afords the corresponding
compound of formula (I) where Rl is (CH3)2CHCOO.
Similarly, use of ethyl chloroformate as the
acylating agent provides the corresponding compound o~
formula (I) where Rl is CH3CH~OCOO and use of fo~mic-
acetic anhydride as acy}ating agent yields (I).where
is ormyloxy.
-38-
EXAMPLE 12
,.,
~ hen the procedure o~ Example 11 is repeated, but
employing the appropriate acid anhydrida, acid chloride,
acid bromide or chloroformate ester as acylating
agen~, the following compoun~s wherein Q is N~C(CH3) =
CHC02CH3 are obtained and hydrolyzed to the corresponding
compounds of the formula wherein Q is ~H2,
R~ CHCONH~< CCH3
~ COOCH20
O O C=O
\ / ' /
O ~ ~ ~COOCH~o
Rl Rl
C~3CH~COO CH30COO
C~3~CH2)2COO C~3CH20COO
C~3(CH2)4COO (CH3~2CHOCOO
(C~3)2C~(C~2)3COO (cH3)3cOcOO
C6H5C (C~3)3CCH~OCOO
3-FC6H4COO C~3(~2)50C
2 C~C6~4COO (C~3)2C~(C~2)30COO
4-n-C4H9C6H4C00 2-BrC6H4COO
2 C~3 6 4 4-BrC6H4CO
4 ~3C6 4 3 n C3H7C6H4
3-C2H50C~H4C00 3-n-C3H7~C6H4COO
4--(CH3)3COC6H4COO ~--IC6H~COO
4-ClC6~C00 3-FC6H4COO
-39-
.
EX~MPLE 13
6-~D-(2-~Benzyloxycarbonylamino]-
2~ hydroxyphenyl~acetamido)]-
penicillanoylvxymethyl
l,l-dioxopenicillanoyloxy-
methyl carbo~ate
(II, Rl = OH, Q = ~HCbz)
To 7.40 g.. (0.010 mole~ tetrabutylammonium 6-CD -
(2-~benzyloxycarbonylam~no]-2-~-hydroxyphenyl]acet-
amido)~penicillanate and 3.81 g. (0.010 mola) bromo-
methyl l,l-dioxopenicillanoyloxymethyl carbonate is
added 50 ml~ dimethylformamide and the mixture is
stixr~Qd for four hours. E~hyl acetate (500 mL.) i~
added and the mix~ure washed in turn with brine,
water, brin~Q again and d~ied over anhydrous sodium
sulfate. ~vaporation o~ solvent in vacuo affords the
crude-product whirh can be purified by chromatography
on silica gel, i~ desired.
Reaction of ~ompounds o the rormula (IV) with
amino-protected penicillin salts of ~he formula (~I)
by ~he above procedure aords products of formula (II)
in like manner, where R , Q, M and X are as derined
below.
~ 2~
--40--
Rl_~>--CHCO~ <C 3 v~< CE3
' COOM ' COOCH2 O
XC~2~=
tVI ) ( IV )
~ ~ r
R~ CHCONH~ CH3
O J COOCH20
0.0 C=O
\5~ CH3
oF~ ~ COOCH2o
(II)
Ri Q M X
H C~jH5CH20CONH ~a
H ~--~02c6EI4c~2ocoNH K B~
EI ~3 K
(n-/ 4~g)~L~ Cl
~o N3 ( n-C4Hg ) 4~ Br
EO p 2C~;EI4 ~2
C~13~00 ~ 4 2 ~ n C4~9 ) 4~ I
6~
EXAM.PLE 14
:
6-~D-~2-Amino-2-~p-hydroxyphenyl)-
acetamido)]penicillanoyloxymethyl
l,l-dioxopenicillanoyloxymethyl carbonate
tI Rl OH)
A mixture of 2.0 g. 6-~D-(2-~benzyloxycarbonyl-
amino~-2-~ -hydro2yphenyl~acetamido)~penicillanoyloxy-
methyl l,l-dioxopenicillanoyloxymethyl carbonate,
50 ml~ dichloromethane, SO ml. isopropanol and 2.0 g.
10% palladium-on-caxbon is hydrogenated at 3-4 atmo-
sphere (3.5-4.0 kg./cm.2) until hydrogen uptake
ceases. An additional 2 g. of catalyst is added and
hydrogenation continued for 30 minutes~ The catalyst
is xemoved by ~ ra~ion and the filtrate evaporat~d
in vacuo to a~ford the product which can be purified,
__
if de~ired, by chromatography on Sephadex L~ 20*.
*A registered trademark of Pharmacia Fine Chemicals,
Piscataway, ~.~T~
~2~
--~2--
EXAMPLE 15
The remaining compounds of formula (II) provided
in Example 13 are converted to the corresponding
alpha-amino compound of formula (I) by the method of
the preceding Example.
R~ 2 ~ ~ C~3
" COOC~20
O O ` C=O
~CCEO~) o/
where Rl is as defined for the starting material of
formula (II)_
-43-
PREPA~ATION A
bis-Ch1oromethyl Carbonate
The method is essentially that of Kling et al.,
Compt. rend. 170, 111-113, 234-236 (1920); Chem.
Abst~., 14, 1304 (1920).
A soLution of 59 ml. dimethyl carbonate in
120 ml~ ~arbon tetrachloride is cooled in an ice bath.
Chlorine gas is bubbled into the solution while
irradiating with a sun lamp un~il most o~ the starting
material is rea~ted. The excess chlorine is displaced
by nitrogen, the solvent evaporated and the residue
was distilled through a short column with ractionating
head at 50 mm. pressure. The desired produc~ boils at
95-100C./50 mm. Yield, 68 g,
.
~L%~6~
_a4_
PREPARATION B
Tetrabutylammonium 6-(2-Benzyloxycarbonyl-
am o-2-~4-hydroxyphenyl~acetamido)penicillanate
To a rapidly stirred mix~ure of l.O g. of 6 (2-
b~n~yloxycarbonylamino-2-~4-hydrox~phenyl]acetamido)-
penicillanic acid, 30 ml~ o~ dichloromethane and 20
ml~ of water was added 40~ aqueous ~etrabutylammonium
hydro~ide until a p~ of 8.0 was obtained. Stirring
was continued or 30 minutes at pH 8.0 and then the
layers were separated. The aqueous layer was extracted
wi~h dichloromethane, and then t~ combined dichloro-
methane soiutions were dried (Na2SO~) and evaporated
in vacuo~ This af~orded l.l g~ o~ t~e title compound~
The ~MR spectrum (in DMSO-d6) ~howed absorptions
at 0~70-lo~O (m, 34~), 2.90-3.50 (m, 8~, 3.93 (5,
lH),. SolO (5~ 2H), 5.23-5.50 (m, 3~1~ 6076 (d, 2~),
7~20 (d, 2H), 7 ao ~s, 5R), 7.76 (d, lH) and 8.6 (d,
lH) ppm_ -
TPtrabutylammonium 6-(2-C4-nitrobenzyloxycarbonyl-
amino~2-C4-hydroxyphenyl]acetamido)penicillanate is
obtained from 6-(2-~4-nitrobenzyloxycarbonylamino~-2-
~4-hydro~yphenyl~acetamido)penicillanic acid and
tetrabutylammonium hydroxide by ~he above method.
Tatrabutylammonium 5-~D-(2-~enzyloxycarbonyl-
amino-2-phe~yl)acetamido]penicillanate, tetrabutyl-
ammon~um-6-~D-2 (4-nitrobenzyloxycarbonylamino)-2-
phenylacetamido~penicillana~e and tetrabutylammQnium
6-~D~ a2ido-2-phenylacetamido)penicillanate ar~
~repared in like manner.
