Note: Descriptions are shown in the official language in which they were submitted.
z
The present invention relates to processes for preparing novel
esters of formula I, to processes for preparing novel salts of the esters
of formula I with pharmaceutically acceptable acids, and to the esters and
salts so produced.
By one aspect of this invention, a process is provided for pro-
ducing a compound of the following formula I
'7
R6 - C 3lYU_~S;
O~\CH2R5
, ~o \
CH
~ N--C _ ~ J
in which Rl and Rz represent Cl 7 alkyl or together with the nitrogcn form
a heterocyclic radical having 4 to 8 carbon atoms; R3 represents hydrogen
or benzyl; R5 represents hydrogen, acyloxy, carbamoyloxy or SR8, where
R8 stands for a heterocyclic ring; R6 represents methyl substituted by
cyano, puridylthio, trifluoromethylthio or by a 5-membered heterocyclic
ring or simultaneously by as-membered herterocyclic ring and by methoximino
or represents benzyl substituted by azido, amino and/or hydroxy or repre-
sents 1,4-cyclohexadienylmethyl substituted by amino; and R7 represents
hydrogen orrnethoxy; and salts thereof with pharmaceutically acceptable
acids. The process cOmprises carrying out one of the following reactions:
(a) reacting an ester of the formula II
- 1- ~
111~4~Z
I N - C = N ~ ~ Il
O N `coocHx
in which Rl, R2 and R3 are as defined above, and X is a lcaving group,
e.g., Cl, Br, I or p-toluenesulphonyloxy, with a salt of the formula III
R~ H S
R6 - ~O~H ~ / ~ lII
O "r-~ HzR5
~no M
in which R5, R6 and R7 have the above meanings and M stands for a cation,
e.g., Na , K , or (C2H5)3NH , to form the selected compound of formula I;
or
(b) converting a compound of formula V
H H
Rg ~
~ N
` COOCH-Cl
wherein, R4
R4 represents hydrogen, alkyl aralkyl or aryl or alkyl, aralky or aryl
substituted with halogen, hydroxy, theo, carboxy or amino groups.in
which R4 is as defined above, and Rg is a protective group, e.g.,
benzyloxycarbonyl, triphenylmethyl, or G ~t~ trichloroethoxycarbonyl,
into the corresponding iodoalkyl ester, e.g., by treatment with sodium
iodide in acetone, the iodoalkyl ester being reacted with a compound of
formula III to yield a compound of formula VI
R6 - CO~H ~ ~
C VI
~ CHR4
Rg - HN
~ N -- /
in which R5 to R7 and Rg have the above defined meanings; and by removal
of the protective group Rg, a compound of formula VII is obtained:
R7
R6 - CONH ~;~ S
2 5
/~ \ VII
O~ O\
2 ~ S ICHR4
O ,"~,
in which R4 to R7 are as defined above, and the compound of formula VII
is thereafter reacted with a reactive derivative of an amide or thioamide
of the formula VIII
X
1~1841Z
1 \ IR3
N - C = Z VIII
in which Rl, R2 and R3 have the above meanings, and Z stands for oxygen
- 3 a -
41Z
or sulphur, to yield the selected compound of formula I; or
(c) reacting an ester of the formula II with a compound of formula III in
which the S atom is oxidized to the ~ S_-~ O grouping to form a compound
of formula IV:
R O
;7 H
6 - CO~ ~ r S~
~ ~ ~ ~ IV
o i, c
,HR4
R ",~'- C ~
N C
0~
in which Rl to R3 and R5 - R7 have the above defined meanings, the com-
pound of formula IV being subjected to a reduction process to form the
selected compound of formula I; and where required, reacting the compound
of formula I obtalned with a pharmaceutically acceptable acid.
The compounds of another aspect of this invention, which are
valuable in human and veterinary prac~ice, have the formula I
7 H
R6 - ~01~
CH2R5
C CH~ I
~ - C = ~
. ,/
o$~
X - 4 -
111~412
in which Rl and R2 represent Cl 7 alkyl or together with the nitrogen form
a heterocyclic radical having 4 to 8 carbon atoms; R3 represeDts hydrogen
or benzyl; R5 represents hydrogen, acyloxy, carbamoyloxy or SR8, where
R8 stands for a heterocyclic ring; R6 represents methyl substituted by
cyano, pyridylthio, trifluoromethylthio or by a 5-membered heterocyclic
ring or simultaneously by a 5-membered heterocyclic ring and by methoxi-
mino or represents benzyl substituted by azido, amino and/or hydroxy or
represents 1,4-cyclohexadienylmethyl substituted by amino; and R7 repre-
sents hydrogen or methoxy; and salts thereof with pharmaceutically
acceptable acids.
In one broad variant, Rl and R2 represent an aliphatic hydrocar-
bon radical in which the carbon chain can be straight or branched, satura-
ted or unsaturated, e.g., methyl, ethyl, propyl, isopropyl, butyl, sec-
butyl, tert-butyl, pentyl, hexyl, dodecyl, allyl, butenyl, pentenyl, pro-
pargyl; a mono- or bicyclic aryl radical, e.g., a phenyl radical or a
naphthyl radical; an aralkyl radical, e.g., a mono- or bicyclic aralkyl
radical, e.g., benzyl, phenylethyl, 1- or 2-naphthylmethyl; a cycloalkyl
or cycloalkylalkyl radical in which the cycloalkyl group can have from 3
to 10 ring members and can be saturated or have one or two double bonds,
e.g., cyclopentyl, cyclohexyl, l-adamantyl, 1-bi~yclo(2.2.2) octyl, cyclo-
pentenyl and cyclohexenyl, cyclopentylmrethyl, cyclohexylmethyl, cyclopen-
tenylethyl, cyclohexenylmethyl, etc.; a heterocyclic radical or a hetero-
cyclically substituted alkyl radical in which the heterocyclic part can
have from 5 to 10 atoms in the ring and can COntain oxygen, sulphur, and/
or nitrogen atoms in all of which the hetero atoms may be placed in any
of the available positions, and such heterocyclic radical optionally being
more or less hydrogenated, e.g., pyridyl, pyrazinyl, pyrimidyl, pyrrolidyl,
piperidyl, morpholinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, thiazinyl,
furyl, thienyl, or quinolyl; wherein each of the radicals Rl
41Z
to R3 can together with one or more of the other radicals represent hetero-
cyclic radicals having from 5 to 8 ring atoms and optionally containing
other hetero atoms in the ring, e.g., S, O and/or N, forming more or less
hydrogenated ring systems, e.g., pyrazolinyl, pyrazolidinyl, 1,2,3,6-tetra-
hydropyridazinyl, 1,4,5,6-tetrahydropyridazinyl, hexahydropyridazinyl,
piperidyl, morpholinyl, hexahydro-lH-azepin-l-yl, or hexahydro-1(2H)-azo-
cinyl. It is preferred that Rl and R2 be Cl 7-alkyl or together with the
nitrogen form a heterocyclic radical having 4 to $ carbon atoms. R3
preferably is hydrogen or benzyl~
It is conceivable with the radicals Rl to R6 may be further sub-
stituted with halogen atoms, and with alkyl, haloalkyl, hydroxy, alkoxy,
aryloxy, alkylthio, arylthio groups and acyl groups, carboxy, N-cyano-
carboximido, carbalkoxy, carbamyl, carbamido, cyano or sulfonyl groups,
azido groups, amino or substituted amino groups.
In yet another variant, Rl and R2 may be selected from the group'
consisting of alkyl, chloroalkyl and hydroxy alkyl having from 1 to 7 car-
bon atoms, alkoxyalkyl having from 2 to 7 carbon atoms, carbalkoxyalkyl
having from 3 to 4 carbon atoms, cyanoal~yl having from 2 to 3 carbon
atoms, and allyl radicals; phenyl-, chlorophenyl-, bromophenyl-, or
phenoxy; carbobenzoxy-substituted methyl and ethyl radicals; cycloalkyl
and cycloalkylmethyl radlcals having f~om 5 to 10 carbon atoms; methyl
and ethyl radicals substituted with a heterocyclic ring selected from the
group consisting of a 5-membered, unsaturated ring having as the hetero
atom an oxygen or sulphur atom, and a 6-membered, unsaturated ring hav-
ing as the hetero atom a nitrogen atom, and Rl and R2 form a straight or
branched alkylene chain which together with the nitrogen atom form a
saturated heterocyclic ring having from 4 to 8 carbon atoms; and wherein
Rl and R2 together with the nitrogen atom form a heterocyclic radical
selected from the group consisting of morpholinyl-4,
341Z
thiomorpholinyl-4, 4~methyl-piperazinyl-1, and 1,2,3,4-tetrahydroisoquino-
lyl-2.
It is also conceivable that R3 can be selected from the group
consisting of hydrogen, and an alkyl or alkenyl radical having from 1 to
7 carbon atoms, a phenyl or benzyl radical, a cycloalkyl or a cycloalkyl-
alkyl radical, the cycloalkyl group of which have from 5 to 6 carbon
atoms and the alkyl group of which have 1 or 2 carbon atoms, a heterocy-
clic radical consisting of a 5 to 6-membered ring containing an oxygen, a
sulphur or nitrogen atom. Further, R3 and Rl when taken together with
the ~-C-atoms between them may represent a heterocyclic fibe-or six-mem-
bered ring system.
In yet another variant, R5 may represent hydrogen, acetoxy,
propionloxy, benzoyloxy, pyridinium, carbamoyloxy, azido, methoxy, or Sr8
in which R8 represents 1-methyl-1~2,3~4-tetrazol-5-yl~ 1,2,3-(lH)-triazol-
5-yl, 1,3,4-thiadiazol-2-yl, or 5-methyl-1,3,4-thiadiazol-2-yl. It is
preferred, however, that R5 be hydrogen, acyloxy, carbamoyloxy or SR8 when
R8 is a heterocyclic ring.
It is conceivable that R6 can represent lower alkyl, unsubsti-
tuted and substituted benzyl,phenoxy-lower alkyl, lower alkyl substituted
with 5- or 6-membered heterocyclic ring system. It is also conceivable
that R6 may represent methyl, benzyl,ror substituted benzyl, e.g., ~ -hy-
droxy, ~-formyloxy, ~-azido, c(-amino, ot-carboxy, ~-sulfo, ~-sul-
foamino benzyl radicals, ~ -amino-p-hydroxybenzyl, and ~-amino-m-chloro-
p-hydroxybenzyl; phenoxymethyl, cyanomethyl, ~ -amino-1,4-cyclohexadienyl-
methyl, trifluormethiomethyl, cyanomethylthiomethyl, heterocyclically
substituted methyl, e.g., 2- and 3-thienylmethyl, ~-amino-2-thienylmethyl,
-amino-3-thienylmethyl, l-(lH)-tetrazolyl-methyl, ~ -methoximino-2-
furyl-methyl, (1,2,3-oxadiazol-5-on-3-yl)-methyl, or 4-pyridylthiomethyl.
It is preferred, however, that R6 be methyl substituted by cyano, pyridyl-
thio,
-- 7 --
11~841Z
trifluormethyl, or by a 5-membered ring or simultaneously by a 5-membered
heterocyclic ring and by methoximinoj or represents benzyl substituted
by azido, amino, and/or hydroxy, or represents 1,4-cyclohexadienylmethyl
substituted by amino~
In a preferred variant, R7 may represent hydrogen or methoxy.
The invention in a further aspect includes all possible isomeric
forms of the compounds of formula I, depending on the different substit-
uents, wherein the 6-aminopenicillanic or 7-aminocephalosporanic acid
moieties have the configurations indicated in the formula I.
As stated above, the invention in still another aspect also
relates to salts of the esters of formula I with pharmaceutically accepta-
ble acids, e.g., hydrochloric and hydrobromic acid, phosphoric acid, sul-
~uric acid, nitric acid, p-toluenesulphonic acid, methanesulfonic acid,
formic acid, acetic acid, propionic acid, citric acid, tartaric acid and
maleic acid. However, salts with other acids which themselves are thera-
peutically active e.g., penicillins, cephalosporins, and fusidic acid,
are also within the scope of aspects of the invention.
As noted above, this invention in other variants also comprises
a process for the preparation of the above-described compounds. In one
embodiment, an ester of the formula II
C = ~ ~ S II
~2 / ,J, ~ ~ ~
` C~OC,HX
in which formula Rl, R2 and R3 are as defined above, and X is a good
leaving group, e.g.~ Cl, Br, I or p-toluenesulphonyloxy, is reacted with
a salt of the formula III
- 1~184~2
R6 ~ CO~'H -7 H S
~00 M
in which R5, R6, and R7 have the above meanings and M stands for a
cation, e~g., Na , K , or (C2H5)3NH~, to form the desired compound of
formula I.
The reaction is preferably performed in a suitable solvent, e.g.,
N,N-dimethylformamide, acetone, or hexamethylphosphoric acid triamide,
usually at a temperature from 0C. to 60C.
In the above embodiment of the process of a variant of this in-
venLion, it is preferred to use a reactant of formula II with X represent-
ing iodine because this results in a product mainly consisting of a compound
in which the double bond of the cephalosporin moiety is in the 3-position.
This is due to the speed of the reaction which doesnot permit the unde-
sired migration of the double bond to the 2-position which would result
in compounds containing an inactive cephalosporin moiety.
In a second embodiment of the process of a variant of the inven-
tion, an ester of the formula II is reacted with a compound of formula II
in which the S atom is oxidized to the ~ S ~ ~0 grouping, to form a co~r
pound of formula IV
_ g _
~lB41Z
S
R6 - co~
N ~ ~ CH2R5 IV
~C o
N ~ R ~ C~
o
in which Rl to R3 and R5 - R7 have the above defined meanings, the com-
pound being subjected to a reduction process to form the desired, selected
compound of formula I.
The reaction is preferably performed in a suitable solvent, e.g.,
methylene chloride or N,N-dimethylformamide,usually at a temperature of
from 0C. to 60C.
The reduction is performed by processes well known from penicil-
lin and cephalosporin chemistry, [see, e.g., J. Org. Chem. 35, (1970),
p, 2433].
ID a third embodiment of therprocess of a variant of this inven-
tion, a compound of formula V
H H H
Rg ~ ~ ~ ~ V
O ~ N -COOC~2Cl
in which Rg is a protective group, e.g~, benzyloxycarbonyl, triphenyl-
methyl, or ~3,/3, ~-trichloroethoxycarbonyl, is converted into the corres-
ponding iodoalkyl ester, e.g., by treatment with sodium iodide in acetone,
the iodoalkyl ester being reacted with a compound of formula III to yield
~ - 10 -
~841~
a compound of formula VI:
R7
R CONH ~ S
N ~ ~
1~ CH2R5 ~II
0~ ~0\
H2
Rg - HN c~ ,S ~
J --- N ~
o
in which R5 to R7 and Rg have the above defined meaning. By removal of
the protectlve group R9 from the compound of formula VI, a compound of
formula VII is obtained:
R7
- - S ~
C Vll
0~ 0
H H Ç~2
H 2~ ~ __ - ~ ~j< o
~ - N - .
0~
in which R5 to R7 are as defined above, and the compound is thereafter
reacted with a reactive derivative of an amide or thioamide of the for-
mula VIII
R~ C = Z \'III
. Il - lla -
8~1Z
in which Rl, R2, and R3 have the above meanings, and Z stands for oxygen
or sulphur, to yield the desired, selec~ed compound of formula I.
As examples of reactive derivatives of a compound of formula
VIII, the following non-limiting types may be given:
L /N ~ C - Cl ] Cl-- iminium chl~rides,
~ F4 iminium e.hers, and
~R~ 3 S - C~5
L i~ s C:~H5 ¦ thioamide acetals
in which Rl, R2, and R3 have the above meanings.
The reactions with the above reactive derivatives are well
known to the man skilled in the art for preparing amidinopenicillanic
acid derivatives, (see, e.g., British Patent No. 1,293,590 which also
describes in detail the meaning of "a reactive derivative of a compound
of formula VIII").
The starting materials of formulae II, III and VIII are known
or may be prepared by procedures analogous to those used for the prepara-
tion of the known compounds.
By a variant of the process described above, Rl and R2 are
methyl or ethyl, or together with the nitrogen atom, form a saturated
heterocyclic ring having 6 carbon atoms; R3 is hydrogen; R5 is selected
from the group consisting of hydrogen, acetoxy, carbamoyloxy, SR8 in which
R8 is selected from the group consisting of 1-methyl-1,2,3,4-tetrazol-5-
yl, 1,2,3-(lH)-triazol-5-yl, 1,3,4-thiadiazol-2-yl and 5-methyl-1,3,4-
thiadiazol-2-yl; and R6 is selccted from the group consisting of benzyl
- 11 b -
J
substituted by ~-hydroxy, c~ -azldo or c~-amido, ~-amino-p-hydroxybenzyl, ..
cyanomethyl, D<-amino-1,4-cyclohexadienylmethyl, trifluormethylthiomethyl,
2-thienylmethyl, l-(lH)-tetrazolyl-methyl, ~<-methoximino-2-furylmethyl
and 4-pyridylthiomethyl.
By another variant, the process includes the step of reacting
the free base with a selected pharmaceutically acceptable acid, wherein
the selected pharmaceutically acceptable acid is selected from the group
consisting of hydrochloric and hydrobromic acid, phosphoric acid, sulfuric
acid, nitric acid, p-toluenesulphonic acid, methanesulfonic acid, formic
acid, acetic acid, propionic acid, citric acid, tartaric acid and maleic
acid; penicillins, cephalosporins, and fusidic acid.
Specific processes within the ambit of preferred embodiments of
this invention to produce the speciic embodiment compounds referred to
include:
(a) the process for preparing 6-[(hexahydro-lH-azepin-l-yl)-
methyleneamino]-penicillanoyloxymethyl 7-[1-(lH)-tetrazolylacetamido3-3-
[(5-methyl-1,3,4-thiadiazol-2-yl)-thiomethyl]-3-cephem-4-carboxylate,
which comprises reacting 6-amino-penicillanoyloxymethyl-7-[1-(lH)-tetra-
zolylacetamido]-3-[(5-methyl-1,3,4-thiadiazol-2-yl)-thiomethyl]-3-cephem-
4-carboxylate with chlorohexamethyleneformiminium chloride including the
later step of reacting the free base with a pharmaceutically acceptable
acid;
(b) the process for preparing 6-~(hexahydro-lH-azepin-l-yl)-
methyleneamino]-penicillanoyloxymethyl 7-(D-~C-aminophcnylacetamido)-
cephalosporanate which comprises hydrogenating 6-[(hexahydro-lH-azepin-
l-yl)-methyleneamino]-penicillanoyloxymethyl 7-(D- ~'-azidophenylacetamido)-
cephalosporanate, including the later step of reacting the free base with
a pharmaceutically acceptable acid;
(c) the process for preparing 6-[(hexahydro-lH-azepin-l-yl)-
~ -- 1 1 c
~, ~
34:1Z
methyleneamino]-penicillanoyloxymethyl 7-(D-ot-azidophenylacetamido)-
cephalosporanate which comprises reacting iodomethyl 6-[(hexahydro-lH-
azepin-l-yl)-methyleneamino]-penicillante with potassium 7-(D-r~ -azido-
phenylacetamido)-cephalosporanate, including the later step of reacting
the free base with a pharmaceutically acceptable acid.
The starting material of formula IV can, e.g., be prepared as
follows: a salt of a N-protected 6-aminopenicillanic acid is reacted
with a compound of formula IX:
~.
(1I R IX
(,1 '
. , ,
d -
i~l841'~
.
where R4 is as previously defined'and Y is a good leaving group, e.g.
Br, I or a p-toluenesulphonyloxy group. The reaction is preferably per-
formed in a sultable solvent,-e.g. dimethylformamide or acetone, and
usually at a temperature between 0C. and 60C.
The'reaction products of formula I can be purified and isolated
in usual manner and may be obtained'either in the free state or in the
form of a salt.
The compounds of aspects of this invention possess valuable
in vivo antibacterial activity and their toxicity is low, even when
administered parenterally.
; Some of the compounds of formula I are absorbed from the
gastrointestinal tract, and these may also be administered enterally.
It has now been shown'that during or after the absorption the
new esters of aspects of the'invention are hydrolyzed by enzymes present
in blood and many tissues with formation of the corresponding free
cephalosporins and amidinopenicillanic acids whereby the simultaneous
presence of these two types of antibiotics in the organism is obtained.
This is particularly advantageous since it is known that cephalosporins
and certain 6-amidino-penicillanic acids show a synergistic effect both
in vitro and in'vivo (E. Grunberg et al.; H.C. New; Presented at the
Fifteenth Interscience Conference on Antimicrobial Agents and Chemo-
therapy in Washington, D.C. September 24-26th, 1975). This is probably
due to the fact that the two types of antibiotics have a different mode
of action. Both interfere with the biosynthesis of the bacterial cell
wall, but the target is different [J.T. Park and L. Burman, Biochem. and
Biophys. Research Comm. 51, 863 (1973)].
The compounds of aspects of the invention are provided in the
form of a composition containing as an active component at least one
member selected from the group consisting of compounds of the formula I
and salts thereof as defined above, together with solid or liquid pharma-
ceutical carriers and/or diluents.
In these compositions, the proportion of therapeutically active
- 12 -
lZ
material to carrier'substance can vary between'1% and 95% by weight.
The compositions can be worked'up to various pharmaceutical forms of
presentation, e.g. tablets, pills, dragees, suppositories, capsules,
sustained-release tablets, suspensions, ointments, injection medicine
and the like containing the compounds of formula I or their atoxic
salts, mixed with carriers and/or diluents.
Pharmaceutical organic or inorganic, solid or liquid carriers
and/or diluents suitable for enteral, parenteral, or topical administra-
tion can be used to make up compositions containing the compounds of
aspects of this invention. Gelatine, lactose, starch, magnesium
stearate, talc, vegetable and animal fats and oils, gum, polyalkylene
glycol, buffers or other known carriers and/or diluents for medicaments
are all suitable.
Furthermore, the compositions may contain other pharmaceuti-
cally active components which can appropriately be administered together
with the present compounds in the treatment of infectious diseases,
such as other suitable antibiotics.
As indicated above, the compounds of aspects of this invention
may be worked up to pharmaceutical forms of presentation including sus-
pensions and non-aqueous ointments and creams. A pharmaceutical prepara-
tian for oral treatment may be in the form of a suspension of one of the
present compounds, the preparation containing from 10 to 100 mg per ml of
a non-aqueous vehicle. A pharmaceutical preparation for topical treat-
ment may be in the form of a non-aqueous ointment or cream containing
one of the present compounds in an amount of from 1/4 to 10 g per 100 g
of preparation.
In the human therapy, the compounds of aspects of this inven-
tion are conveniently administered (to adults) in dosage units containing
not less than 25 mg and up to 1000 mg, preferably from 100 to 500 mg,
calculated as the compound of formula I.
By the'term "dosage unit" is meant a unitary, i.e. a single
dose which is capable of being administered to a patient, and which may
41Z
be readily handled and packed, remaining as a physically stable unit
dose comprising either the'active material as such or a mixture of it
with solid or liquid pharmaceutical diluents or carriers.
In the form of a dosage unit, the compound may be administered
once or more times a day at appropriate intervals, always depending,
however', on the condition of the patient, and in accordance with the
prescription made by the medical practitioner.
Thus a daily dose will preferably be an amount of from 1 to 3
g of a compound of formula I or an equivalent amount of a salt thereof
as defined before.
If the composition is to be injected, a sealed ampoule, a
vial or a similar container may be provided containing a parenterally
acceptable aqueous or oily injectable solution or dispersion of the
active material as the dosage unit.
The parenteral preparations are in particular useful in the
treatment of conditions in which a quick response to the treatment is
desirable.~ In the continuous therapy of patients suffering from infec-
tions diseases, the tablets or capsules may be the appropriate form of
pharmaceutical preparation.
In the treatment of infectious diseases, such tablets may'
advantageously contain other'active components, as mentioned hereinbefore.
The compounds of aspects of this invention are useful in
treating patients suffering from infectious diseases. The method of use
compri8es administering to adult patients from 0.5 g to 5 g per day,
preferably from 1 to 3 g per day, of a compound of the formula I or an
equivalent amount of a salt as defined before of a compound of the
formula I. Preferably, the compound is given in the form of the dosage
units aforesaid.
The invention will be further described in the following
~xamples whlch are not to be construed as limiting the invention, but
which are given to provide Examples of variants and embodiments of this
invention. '
341Z
Example 1
'Iodomethyl 6-[(hexahydrd-lH-aæepin-l-yl~-methyleneamino]-penicillanate.
To a solution of chloromethyl 6-[(hexahydro-lH-azepin-l-yl)-
methyleneamlno]-penicillanate '(0.5 g~ in acetone (4 ml) was added sodium
iodide (0.8 g). The'mixture was stirred'at 40C. for 1 hour, cooled and
filtered. The filtrate was diluted with either and washed repeatedly
with water. The organic phase was dried and evaporated to leave the
desired compound as an oil, which was used in the next step without
purification.
The NMR-spectrum (CDC13) showed signals at ~= 1.55 (s),
1.70 (s), 4.37 (s), 1.5-2.0 (m), 3.3-3.6 (m), 5.20 (dd, J=l, J=4),
5.50 (d, J=4), 5.90 (d, J=6), 6.03 (d, J=6) and 7.68 (s) ppm.
TMS was used as internal standard.
Example 2
6-[(Hexahydro-lH-azepin-l-yl)-methyleneamino]-penicillanoyloxymethyl
7-(D-~ -a~idophenylacetamido)-cephalosporanate, hydrochloride.
To~n ice-cold solution of iodomethyl 6-[(hexahydro-lH-azepin-
_ l-yl)-methyleneamino]-penicillanate (1.19 g) in dimethylformamide (20 ml),
potassium 7-(D- ~-azidophenylacetamido)-cephalosporanate (1.5 g) was
added. After stirrin8 for 10 minutes'the'mixture was diluted with ethyl
acetate and washed repeatedly with water. The organic phase and water
was stirred'while hydrochloric acid was added to pH = 4. The aqueous
phase containing some impurities was discarded and the organic phase was
stirred'with watçr while hydrochloric acid was added to pH = 2.5. The
aqueous phase was freeze-dried to yield the title compound as a colour-
less powder.
The IR-spectrum (KBr) showed strong bands at 2110, 1780, 1740
and 1680 cm 1.
The NMR-spectrum (CD30D, TMS as internal reference) showed
peaks at ~ - 1.58 (s), 1.73 (8), 1.4-2.2 (m), 2.05 (s), 3.55 (m), 3.6-4.0
(m), 4.63 '(s), 4.82 (d, J-14), 5.08 (d, J=14), 5.10 (d, J=5), 5.15 (s),
5.4-5.9 (m), 5;99 ~s), 7;43 '(s) and 8.17 ~bs) ppm.
- 15 -
341Z
' EXample 3
6-[(Hexahydro-lH-azepin-l-yl)-methyleneamino]-penicillanoyloxymethyl
.
''7-(D-~'-aminophenylacetamido~-cephalosporanate, dihydrochloride.
To a stirred solution of the compared prepared according to
Example 2 (0.5 g) in water'(25 ml), 10% Pd-C (0.2 g) was added. Hydrogen
was bubbled through the mixture while a pH-value of 3 was maintained by
the addition of hydrochloric acid. When the consumption of acid stopped
the catalyst was filtered'off and the filtrate was freeze-dried to yield
the title compound as a colourless powder.
The IR-spectrum (KBr) showed strong bands at 1775, 1740 and
1685 cm 1.
The NMR-spectrum (CD30D, TMS as internal reference) showed
peaks at ~ = 1.57 (s), 1.73 '(s), 1.5-2.2'(m), 2.03 (s), 3.53 (m), 3.5-4.0
(m), 4.62 (6), 5.11 (s), 4.8-5.3 (m), 5.5-6.0 (m), 5.98 (s), 7.50 (s) and
8.20 (s) ppm.
''Examples"4'-'15
By following the procedures'described in Examples 2 andtor 3,
but replacing potassium 7-(D-~ -azidophenylacetamido)-cephalosporanate
with a compound of formula III, the compounds of formula I in Table I are
prepared in the form of their hydrochlorides.
- 16 -
8412
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Example 16
' Chl~romethyl'6-(N-triphenylmethylamino)-penicillanate
To a solution of 6-(N-triphenylmethylamino)-penicillanic acid
(5 g) in dimethylformamide '(50 ml) was added triethylamine (1.53 ml)
and chloroiodo-methane (5 ml). 'After stirring at room temperature for
3 hours, the mixture was diluted'with ethyl acetate (200 ml) and washed
with water t4 x 50 ml). The organic phase was dried and evaporated to
leave chloromethyl 6-(N-triphenylmethylamino)-penicillanate as a yellow
oil.
The NMR-spectrum (CDC13) showed signals at ~ = 1.38 (s),
1.58 (s~, 3.18 (d, J=ll), 4.38 (s), 4.55 (m), 5.60 (d, J=6), 5.75 (d,
J=6) and 7.1-7.7 (m) ppm. TMS was used as internal standard.
- Example 17
- Iodomethyl:6-(N-triphenylmethylamino)-penicillanate.
To a solution of chloromethyl 6-(N-triphenylmethylamino)-
penicillanate (0.65 g) in acetone (4 ml) was added sodium iodide (0.8 g).
The mixture was stirred at 40C. for 3 hours, cooled and the precipi-
tated sodium chloride filtered'off. The filtrate was diluted with
ether and washed repeatedly with water. The organic phase was dried
and evaporated to leave a yellow oil, which was used in the next step
without purification.
The NMR-spectrum (CDC13) showed signals at J = 1.38 (s),
1.57 (s), 3.15 (m), 4.33 (g), 4.45 (m), 5.77 (d, J=6), 5.92 (d, J=6)
and 7.1-7.7 (m) ppm.
TMS was used as internal standard.
Example 18
6-(N-triphenylmethylamino)-penicillanoyloxymethyl 7-[1-(lH)-tetrazolyl-
acetamido]-3-[(5-methyl-1,3,4-thiadiazol-2-yl)-thiomethyl]-3-cephem-4-
carboxylaté. _ '
To an ice-cold solution of iodomethyl 6-(N-triphenylmethylamino)-
penicillanate (0.29 g) in dimethylformamide (6 ml), sodium 7-[1-(lH)-
tetrazolylacetamido3-3-{(5-methyl-1,3,4-thiadiazol-2-yl)-thiomethyl]-3-
L1341Z
cephem-4-carboxylate (cefazoline sodium) (0.23 g) was added. After
stirring for 10 minutes'the mixture was diluted'with ehtyl acetate and
washed several times with water. The organic phase was dried and
evaporated to leave the desired'compound as an oil.
Thin-layer chromatography (SiO2-plates, solvent system: ethyl
acetate) showed after 2 consecutive runs one spot with a Rf-value of 0.53.
Example l9
6-Amino-penicillanoyloxymethyl 7-[1-(lH)-tetrazolylacetamido]-3-[(5-
--methyl-1,3,4-thiadiazol-2-yl~-thiomethyl]-3-cephem-4-carboxylate.
A solution of the'compound prepared in Example 18 (0.1 g) in
acetone (2 ml) was treated with p-toluenesulphonic acid (21 mg) at room
temperature for 15 minutes. Then aqueous sodium bicarbonate and ethyl
acetate was added. The organic phase was separated and washed with water,
dried and evaporated to yield an oil which was used in the next step
without further purification.
Example 20
6~[(Hexahydro-lH-azepin-l-yl)-methyleneamino]-penicillanoyloxymethyl
7-[1-(lH)-tetrazolylacetamido]-3-[(5-methyl-1,3,4-thiadiazol-2-yl)-
thiomethyl]-3-cephem-4-carboxylate,'hydrochloride.
A solution of 6-amino-penicillanoy~oxymethyl 7-[1-(lH)-tetra-
zolylacetamido]-3-[(5-methyl-1,3,4-thiadiazol-2-yl)-thiomethyl]-3-cephem-
4-carboxylate (1.75 g) in alcohol-free chloroform was cooled to -40C.
and pyridine'(0.43 ml) was added. A solution of chlorohexamethylene-
ormiminium chloride (0.95 g) in alcohol-free chloroform was added drop-
wise and the mixture was stirred for 0.5 hour at -20C. After stirring
for a further 15 minutes at 0C., the solvent was evaporated and the
residue was treated with ethyl acetate and aqueous sodium bicarbonate.
The organic phase was separated and washed with water. The organic phase
was stirred with water while hydrochloric acid was added to pH=2.5. The
aqueous phase was separated and freeze-dried to yield the desired compound
which was identical with that prepared in Example 4.
The compound was homogeneous according to thin-layer
- 19 -
341Z
chromatography and a bioautogram showed only one active compound.
The NMR-spectrum (CD30D, TMS as internal reference) showed
peaks at ~ = 1.60 (s), 1.73 ~s), 1.4-2.1 (m), 2.75 (s), 3.60 (m),
3.6-3.9 (m), 4.65 (s), 4.20 (d, J=14), 4.65 (d, J=14), 5.16 (d, J=6),
5.46 (m), 5.4-5.9 (m), 6.02 (m), 8.24 (s) and 9.18 ~s) ppm.
Examples 21 - 26
By following the procedures described in Examples 18 - 20 but
replacing cefazoline sodium with a compound of formula III and chloro-
hexamethyleneformininium chloride with a reactive derivative of a com-
pound of formula VIII the compounds of formula I in Table II and TableIII are prepared in the form of their hydrochlorides.
- 2Q -
457
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Example 27
Enzymatic hydrolysis of 6-~(hexahydro-lH-azepin-l-yl)-methyleneamino]-
peniciilanoyloxymethyl 7-[1-(lH)-tetrazolylacetamido]-3-[(5-methyl-
1,3,4-thiadiazol-2-yl)-thiomethyl]-3-cephem-4-carboxylate, hydrochlor-
ide (A)- `
(A) was added to heparinized human blood at a concentration
of 0.5 mg/ml and the mixture was incubated at 37C. for 30 minutes.
After ultrafiltration the ultrafiltrate was subjected to paper chromato-
graphy together with standard solutions of cefazoline sodium (B),
6-[(hexahydro-lH-azepin-l-yl)-methyleneamino]-penicillanic acid (C) and
(A).
, Solvent systems: a) butanol: acetic acid: water (4:1:5)
b) ethyl acetate (saturated with acetate
buffer, pH = 5.6)
The paper chromatograms from a) and b) were placed on agar
plates seeded with E. coli and Staph. aureus respectîvely. After
incubation overnight at 37C. both plates showed no zone of inhibition
in the ultrafiltrate cor~esponding to (A).
The E. coli plate showed a zone of inhibition in the ultra-
filtrate with the same Rf-value as that of (C). The Staph. plate showed
a zone of inhibition in the ultrafiltrate with the same Rf-value as that
Of ~B)-
These result6 indicate that the title compound (A) is hydro-
lyzed into (B) and (C) by enzymes present in human blood.
-Example 28
By using the technique described in Example 27 the compounds
described in Examples 2 - 15 and 20 - 26 under the influence of enzymes
present in human blood are cleaved into the corresponding free
cephalo~porins and amidinopenicillanic acids.
Example 29
Ingredients for 1000 tablets:
6-[(Hexahydro-lH-azepin-l-yl)-tetrazolylacetamido]-3-[~5-methyl-
- 22 -
34~;~
1,3,4-thiadiazol-2-yl)-thiomethyl]-3-cephem-4-carboxy]ate, hydro-
chloride (A) ................................................. 250 g
Palyvinylpyrrolidone .......................................... 10 g
Isopropanol ................................................. 100 ml
Microcrystalline cellulose .................... ......... 165 g
Corn starch .................................... ......... ...... 71 g
Magnesium stearate ............................. ......... ....... 4 g
Sieve A through a screen with 1 mm openings and wet the po~de~
with a solution of polyvinylpyrrolidone into isopropanol. Dry the
moist mass at 30~C. and pass it through a sieve with 0.7 mm openings.
Mix the granules with microcrystalline cellulose, corn starch
and magnesium stearate.
Press tablets of 0.500 g weight using 12 mm punches and dies
to yield tab]ets each containing 250 mg of A.
- 23 -
