Novel 7-amino-thiazolyl-acetamido-cephalosporanic acid compounds of the formula ##STR1## wherein R is selected from the group consisting of hydrogen and a group easily removable by acid hydrolysis or hydrogenolysis, R.sub.1 is selected from the group consisting of alkyl of 1 to 4 carbon atoms, a 5 member heterocyclic ring and a 5 member heterocyclic ring containing a ketone group and A is selected from the group consisting of hydrogen, alkali metal and equivalents of alkaline earth metals, magnesium and organic amine having antibiotic activity against gram negative and gram positive bacteria and their preparation and novel intermediates therefore. PAC STATE OF THE ART French Pat. No. 2,255,077 of Takeda describes antibacterial cephalosporins having a different substitutent in the 3-position.

OBJECTS OF THE INVENTION 
It is an object of the invention to provide the novel cephalosporanic acid 
compounds of formula I. 
It is another object of the invention to provide a novel process for the 
preparation of the compounds of formula I and novel intermediates 
therefore. 
It is a further object of the invention to provide novel antibiotic 
compositions and a novel method of combatting bacterial infections in 
warm-blooded animals, including humans. 
These and other objects and advantages of the invention will become obvious 
from the following detailed description. 
THE INVENTION 
The novel final products of the invention are 
7-amino-thiazolyl-acetamido-cephalosporanic acid compounds of the formula 
##STR2## 
wherein R is selected from the group consisting of hydrogen and a group 
easily removable by acid hydrolysis or hydrogenolysis, R.sub.1 is selected 
from the group consisting of alkyl of 1 to 4 carbon atoms, a 5 member 
heterocyclic ring and a 5 member heterocyclic ring containing a ketone 
group and A is selected from the group consisting of hydrogen, alkali 
metal and equivalents of alkaline earth metals, magnesium and an organic 
amine. 
Groups that are easily removable by acid hydrolysis or hydrogenolysis are 
well known in cephalosporin chemistry and include tert.-butoxy carbonyl, 
trityl, benzyl, dibenzyl, trichloroethyl, carbobenzyloxy, formyl and 
phthaloyl. 
Examples of R.sub.1 are methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, 
tert.-butyl, furyl, thiazolyl, oxothiazolinyl, isothiazolyl, oxazolyl, 
imidazolyl, diazolyl, thiadiazolyl and tetrazolyl. Examples of A are 
hydrogen, sodium, potassium, lithium, calcium and magnesium and organic 
amines such as triethylamine, trimethylamine, methylamine, propylamine, 
N,N-dimethyl ethanolamine or tris(hydroxymethyl)methylamine. 
Among the preferred compounds of formula I, R is tert.-butoxycarbonyl, 
trityl, dibenzyl, trichloroethyl and carbobenzyloxy. Preferably R is 
hydrogen or trityl and R.sub.1 is methyl, ethyl, furyl or 2-oxo (3H) 
thiazolin-4-yl and A is hydrogen. The most preferred compound is 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-acetylthiomethyl-ceph-3-eme-4-carb 
oxylic acid. 
The novel products can exist in the form of formula I or in the following 
formula 
##STR3## 
wherein R, R.sub.1 and A have the above definitions. 
The process of the invention for the production of compounds of formula I 
comprises reacting a compound of the formula 
##STR4## 
wherein R.sub.1 has the above definition with an acid of the formula 
##STR5## 
or a functional derivative thereof wherein R' is a group easily removable 
by acid hydrolysis or hydrogenolysis to obtain a compound of the formula 
##STR6## 
which is a compound of formula I wherein R is other than hydrogen and A is 
hydrogen and the said product may be treated with an acid media or 
subjected to hydrogenolysis to obtain the corresponding compound of 
formula I.sub.b which is a compound of formula I, wherein R is hydrogen 
and if desired the product Ia or Ib may be salified with a mineral or 
organic base or amine to obtain the compounds of formula I where A is 
other than hydrogen. 
In a modification of the process of the invention, a compound of the 
formula 
##STR7## 
wherein R has the above definition and A' is hydrogen or an alkali metal 
is reacted with a compound of the formula 
##STR8## 
wherein R.sub.1 has the above definition and B is hydrogen or an alkali 
metal to obtain the corresponding compound of formula I wherein A is 
hydrogen or an alkali metal which if salified may be treated with an acid 
to obtain the free acid. 
In a preferred mode of the first process described above, the compound of 
formula II is reacted with a functional derivative of the acid of formula 
III such as the acid chloride or acid anhydride. The acid anhydride may be 
formed in situ by action of isobutyl chloroformate on the free acid. Other 
acid halides or other acid anhydrides formed in situ by reaction with 
other alkyl chloroformates, a dialkylcarbodiimide or a 
dicycloalkylcarbodiimide such as dicyclohexylcarbodiimide can be used. 
Equally useful are other acid derivatives such as the azide, amide or 
ester of the acid formed such as with hydroxy succinimide, p-nitrophenol 
or 2,4-dinitrophenol. 
If the compound of formula II is reacted with the acid halide of the acid 
of formula III or an anhydride formed with isobutyl chloroformate, the 
reaction is preferably effected in the presence of a basic agent such as 
an alkali metal carbonate or a tertiary organic base such as 
N-methylmorpholine, pyridine or a trialkylamine such as triethylamine. 
The acid hydrolysis agent to form the compounds of formula Ib may be an 
acid such as trifluoroacetic acid, formic acid or acetic acid. These acids 
can be used in an anhydrous or aqueous medium. A preferred agent for 
hydrogenolysis is zinc-acetic acid system. The preferred hydrolysis agent 
to remove trityl or tert.-butoxycarbonyl groups is anhydrous 
trifluoroacetic acid or aqueous formic or acetic acids. The zinc-acetic 
acid system is preferably used to remove the trichloroethyl group and the 
catalytic hydrogenation is preferably used to remove benzyl, dibenzyl or 
carbobenzyloxy groups. 
The reaction of the products of formulae IV and V is preferably effected in 
water or a water-acetone mixture but equally useful are other aqueous 
solvents such as water-dioxane, water-tetrahydrofuran or water-ethanol. 
In the case of the product of formula IV where A' is hydrogen or the 
product of formula V where B is hydrogen or the case where A' and B are 
both hydrogen, the preferred mode converts the acids in situ to the alkali 
metal salt of formula IV and/or V. The preferred alkali metals are sodium 
or potassium and they are preferably used in the form of their 
bicarbonates. However, other alkaline bases such as sodium hydroxide, 
potassium hydroxide or sodium or potassium carbonate are equally useful. 
The reaction of the products of formulae IV and V is preferably effected in 
the presence of a buffering agent to keep the pH of the medium 
substantially neutral. Such a buffering agent is monosodium 
phosphate-sodium bicarbonate. To form the free acid of formula I, the 
alkali metal salts are preferably treated with acetic acid, but other 
mineral or organic acids such as hydrochloric acid, sulfuric acid, formic 
acid, oxalic acid or trifluoroacetic acid may also be used. 
In the case where the compound of formula IV in which A' is an alkali metal 
and R is hydrogen and the compound of formula V in the alkali metal salt 
form are reacted in stoichiometric amounts, the acetic acid freed during 
the reaction can permit the products of formula Ib to crystallize in the 
form of an internal salt, thus rendering unnecessary the use of an acid as 
discussed above. 
The salification of free acids of formula I may be effected by known 
methods such as reaction of the acids with a mineral base like sodium or 
potassium hydroxide or sodium bicarbonate or an organic base such as 
triethylamine. The salification may be effected in one or more solvents 
such as water, ether, ethanol or acetone. 
The novel antibiotic compositions of the invention are comprised of an 
antibiotically effective amount of at least one compound of formula I and 
an inert pharmaceutical carrier. The compositions may be in the form of 
tablets, dragees, gelules, granules, suppositories, injectable solutions 
or suspensions, pomades, creams and gels prepared in the usual manner. 
Examples of suitable excipients or carriers are talc, arabic gum, lactose, 
starch, magnesium stearate, cacao butter, aqueous and non-aqueous 
vehicles, fatty bodies of animal or vegetable origin, paraffinic 
derivatives, glycols, preservatives and diverse wetting agents, 
dispersants and emulsifiers. 
The compositions are effective against gram positive bacteria such as 
staphylococcus, streptococcus and especially penicillin-resistant 
staphylococcus as well as against gram negative bacteria especially 
coliform bacteria, proteus and klebsiellae. 
The compositions are therefore useful in the treatment of staphylococcia 
such as septicemia of staphylococcus, staphylococcia malignant on the face 
or skin, pyodermititis, septic or suppurantes sores, antrax, phlegmons, 
eresipels, acute primitive or post-grip staphylococcia, bromchopneumonia 
or pulmonary suppurations. They are equally useful for the treatment of 
collibacillosis and associated infections, infections of Proteus and 
Klebsiella and other affections caused by gram negative bacteria. 
The novel method of combatting bacterial infections in warm-blooded 
animals, including humans, comprises administering to warm-blooded animals 
an antibacterially effective amount of at least one compound of formula I. 
The compounds may be administered orally, rectally, parenterally or 
locally by topical application to the skin or mucous. The usual effective 
daily dose is 5 to 80 mg/kg depending on the specific compound, the 
bacteria and the route of administration. 
The starting materials of formula II are prepared by reacting a compound of 
formula V with 7-amino-cephalosporanic acid. The products of formula II 
wherein R.sub.1 is alkyl of 1 to 4 carbon atoms and especially 
7-amino-3-acetylthiomethylceph-3-eme-4-carboxylic acid are novel products. 
The products of formula III are prepared by classical reactions of 
protection of amine functions applied to 2-amino-4-thiazolyl-acetic acid 
or its esters. The products of formula IV are prepared by reacting a 
compound of formula III with 7-amino-cephalosporanic acid followed by acid 
hydrolysis. 
The products of formula V that are not known may be prepared by the 
reaction of sodium sulfhydrate with the acid chloride of the formula 
R.sub.1 COCl according to the method described in J. Antibiotics, 27-8-577 
(1974). 
The products of Example 4 may exist in the form indicated or in the form of 
7-2-(2-amino-4-thiazolyl)acetamido!-3-{(2-hydroxy-4-thiazolyl) 
carbonyl}-thiomethyl!ceph-3-eme-4-carboxylic acid. 
In the following examples there are described several preferred embodiments 
to illustrate the invention. However, it is to be understood that the 
invention is not intended to be limited to the specific embodiments.

EXAMPLE 1 
7-2-(2-tritylamino-4-thiazolyl)-acetamido!-3-acetylthiomethyl-ceph-3-eme-4 
-carboxylic acid 
STEP A: 2-tritylamino-4-thiazolyl-acetic acid 
A mixture of 930 mg of ethyl 2-amino-4-thiazolyl-acetate, 25 ml of dry 
chloroform, 0.8 ml of triethylamine and 1.65 g of trityl chloride was 
stirred for 3 hours and then 3 ml of N hydrochloric acid and 5 ml of water 
were added thereto. The mixture was stirred and decanted and 5 ml of N 
hydrochloric acid and 5 ml of water were again added. The mixture was 
decanted and the organic phase was dried and evaporated to dryness. The 
residue was added to 10 ml of dioxane and 6 ml of N sodium hydroxide and 
the mixture was stirred at 50.degree. C and then stood overnight at room 
temperature. The mixture was evaporated to dryness and the residue was 
diluted with water. The solution was washed with ether and was acidified 
with 0.5 ml of acetic acid. The mixture was allowed to crystallize and was 
then vacuum filtered to obtain 1.33 g of 2-tritylamino-4-thiazolyl-acetic 
acid which after empasting with ether melted at 220.degree. C. 
STEP B: 
7-2-(2-tritylamino-4-thiazolyl)-acetamido!-3-acetoxymethyl-ceph-3-eme-4-c 
arboxylic acid 
A mixture of 801 mg of 2-tritylamino-4-thiazolyl-acetic acid, 10 ml of dry 
tetrahydrofuran and 2 ml of 1M solution of N-methyl-morpholine in 
tetrahydrofuran was stirred and cooled to -20.degree. C and 2 ml of a 1M 
solution of isobutyl chloroformate in tetrahydrofuran were slowly added 
thereto. The mixture was stirred and then a solution of 544 mg of 
7-amino-cephalosporanic acid in 24 ml of a 1 M solution of 
N-methyl-morpholine in tetrahydrofuran and 10 ml of water was added 
thereto. The mixture was stirred with reheating and the solvent was 
evaporated. The residue was diluted with water and 2 ml of 2N hydrochloric 
acid were added. The mixture was vacuum filtered to obtain 1.16 g of 
7-2-(2-tritylamino-4-thiazolyl)-acetamido!-3-acetoxy 
methyl-ceph-3-eme-4-carboxylic acid. 
STEP C: 7-2-(2-tritylamino-4-thiazolyl)-acetamido!-3-acetyl 
thiomethyl-ceph-3-eme-4-carboxylic acid 
A mixture of 1.63 of the product of Step B, 0.21 g of sodium bicarbonate, 
0.66 g of potassium thioacetate, 10 ml of water and 5 ml of acetone was 
heated at 90.degree. C for 21/2 hours and was then acidified with 1 ml of 
acetic acid. The mixture was vacuum filtered and the solid product was 
washed and chromatographed over silica gel. Elution with a 1-1 methylene 
chloride-ether mixture and then with a 5-1-4 acetone-water-ether mixture 
yielded 830 mg of 
7-2-(2-tritylamino-4-thiazolyl)-acetamido!-3-acetylthiomethyl-ceph-3-eme- 
4-carboxylic acid. 
EXAMPLE 2 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-acetylthiomethyl-ceph-3-eme-4-carbo 
xylic acid 
The product of Example 1 in 50% aqueous formic acid was heated at 
60.degree. C for 15 minutes and the formic acid was evaporated. The 
residue was taken up in acetone and the mixture was vacuum filtered to 
obtain 325 mg of product. The latter was dissolved in 2.6 ml of acetone 
containing 20% of water and 0.3 ml of 2N hydrochloric acid and the mixture 
was vacuum filtered. 3 drops of pyridine and 1 ml of acetone were added to 
the filtrate to obtain 0.228 mg of purified 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-acetylthiomethyl-ceph-3-eme-4-carb 
oxylic acid. 
Analysis: C.sub.15 H.sub.16 O.sub.5 N.sub.4 S.sub.3 : Calculated: %C, 
42.04; %H, 3.76; %N, 13.07; %S, 22.44. Found: %C, 41.7; %H, 3.9; %N, 12.8; 
%S, 20.9. 
EXAMPLE 3 
7-2-(2-tritylamino-4-thiazolyl)-acetamido!-3-acetylthiomethyl-ceph-3-eme-4 
-carboxylic acid 
STEP A: 7amino-3-acetylthiomethyl-ceph-3-eme-4-carboxylic acid 
A mixture of 5.44 g of 7-amino-cephalosporanic acid and 50 ml of water with 
1% hydroquinone was stirred under an inert atmosphere and 1.7 g of sodium 
bicarbonate were added thereto. After dissolution occurred, 3 g of 
potassium thioacetate were added thereto and the mixture was heated for 3 
hours at 60.degree. C and then was cooled. The mixture was acidified with 
acetic acid and stirred at room temperature and then vacuum filtered. The 
recovered product was washed and dried to obtain 4.9 g of 
7-amino-3-acetylthiomethyl-ceph-3-eme-4-carboxylic acid. 
STEP B: 
7-2-(2-tritylamino-4-thiazolyl)-acetamido!-3-acetylthiomethyl-ceph-3-eme- 
4-carboxylic acid 
A mixture of 9.9 g of 2-tritylamino-4-thiazolylacetic acid, 100 ml of 
tetrahydrofuran and 2.7 ml of N-methylmorpholine was stirred under an 
inert gas for 15 minutes at room temperature and was then cooled to 
-15.degree. C after which 3.15 ml of isobutyl chloroformate were added. 
The mixture was stirred for 5 minutes at -10.degree. to -15.degree. C and 
then 6.5 g of the product of Step A, 65 ml of water and 3.15 ml of 
triethylamine were added over 2 minutes. The mixture was stirred for 11/2 
hours at room temperature and the tetrahydrofuran was removed. The mixture 
was acidified and was extracted with methylene chloride. The mixture was 
vacuum filtered and the organic phase was washed and evaporated to 
dryness. The powdery residue was triturated with ether and dried to obtain 
15.2 g of raw product. 6.7 g of of the said product were dissolved in 
methylene chloride and ethyl acetate was added thereto. The insoluble 
phase was removed and the solution was treated with carbon black and was 
vacuum filtered. The recovered product was rinsed and dried to obtain 5.1 
g of 
7-2-(2-tritylamino-4-thiazolyl)-acetamido!-3-acetylthiomethyl-ceph-3-eme- 
4-carboxylic acid. 
EXAMPLE 4 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-{(2-oxo-(3H)-thiazolin-4-yl)-carbo 
nyl}-thiomethyl!-ceph-3-eme-4-carboxylic acid 
STEP A: 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-acetoxymethyl-ceph-3-eme-4-carboxy 
lic acid 
A mixture of 351 mg of 
7-2-(2-tritylamino-4-thiazolyl)-acetamido!-3-acetoxymethyl-ceph-3-eme-4-c 
arboxylic acid 0.44 ml of acetic acid and 0.22 ml of water was heated to 
60.degree. C and was stirred for 40 minutes and then cooled. The mixture 
was diluted with acetone and then with ether and was vacuum filtered. The 
precipitate was dried to obtain 171 mg of raw product. 
470 mg of the said product were dissolved in 5 ml of a refluxing 1-1 
mixture of ethanol and water and the mixture was vacuum filtered. The 
filtrate was cooled and vacuum filtered again to obtain a first yield of 
170 mg. The filtrate of the first crop was combined with the first 
insolubles to obtain a second yield of 107 mg for a total yield of 277 mg 
of 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-acetoxymethyl-ceph-3-eme-4-carboxy 
lic acid. 
STEP B: (2-oxo-(3H)-thiazolin-4-yl)-thiocarboxylic acid 
1.635 g of (2-oxo-(3H)-thiazolin-4-yl)-carboxylic acid chloride were added 
to a mixture of 1.6 g of 70% sodium sulfhydrate, 16 ml of ethanol and 2.5 
ml of water at 10.degree. C and the mixture was stirred for an hour. The 
ethanol was evaporated and water was added to the reaction mixture. 0.8ml 
of concentrated hydrochloric acid were added with stirring and the mixture 
was vacuum filtered. The recovered product was rinsed and dried to obtain 
1.46 g of (2-oxo-(3H)-thiazolin-4-yl)-thiocarboxylic acid. 
STEP C: 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-{(2-oxo-(3H)-thiazolin-4yl)-carbo 
nyl}thiomethyl!-ceph-3-eme-4-carboxylic acid 
A mixture of 0.825 g of the product of Step A, 4 ml of distilled water, 0.4 
g of the product of Step B, 0.39 g of monosodium phosphate and 0.21 g of 
sodium bicarbonate was stirred for 5 hours at 50.degree. C and was then 
cooled. Acetic acid was added thereto to adjust the pH to 4 to 5 and the 
mixture was vacuum filtered to obtain after drying 0.56 g of raw product. 
The latter was dissolved in 5 ml of water containing an equivalent of 
sodium bicarbonate and the solution was passed through an ion exchange 
resin column. The column was eluted with water and the product was 
recovered with water containing 10% isopropanol. The fractions were 
combined and acidified with acetic acid to obtain 0.12 g of 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-{(2-oxo-(3H)-thiazolin-4-yl)-carb 
onyl}thiomethyl!-ceph-3-eme-4-carboxylic acid. 
UV spectrum (ethanol containing 10% HCl): Max. at 258 nm, .epsilon. = 
16,350; Max. at 307 nm, .epsilon. = 10,350; 
Infrared spectrum (Nujol): 1722 cm.sup.-1 (.beta.-lactam); 1682-1638 
cm.sup.-1 (secondary amide 
##STR9## 
EXAMPLE 5 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-{(2-furyl)-carbonyl}thiomethyl!-ce 
ph-3-eme-4-carboxylic acid 
STEP A: sodium 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-acetoxymethyl-ceph-3-eme-4-carboxy 
late 
A mixture of 2.26 g of 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-acetoxymethyl-ceph-3-eme-4-carboxy 
lic acid, 420 mg of sodium bicarbonate, 10 ml of water and 0.5 g of 
activated carbon was stirred for 3 minutes at 50.degree. C and was then 
vacuum filtered. The filtrate was washed with acetone and concentrated to 
5 ml and 50 ml of acetone were added. The mixture was heated to 60.degree. 
C to cause crystallization and was vacuum filtered to obtain 1.745 g of 
sodium 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-acetoxymethyl-ceph-3-eme-4-carboxy 
late with a specific rotation of .alpha.!.sub.D = + 120.degree. (C = 1% in 
water). 
STEP B: 7-2-(2-amino-4-thiazolyl)-acetamido!-3-{(2-furyl) 
carbonyl}thiomethyl!-ceph-3-eme-4-carboxylic acid 
A mixture of 0.25 g of 2-furyl-thioacetic acid, 4 ml of distilled water, 
0.17 mg of sodium bicarbonate and 0.435 mg of the salt of Step A was held 
at 50.degree. C for 5 hours and was then cooled to room temperature. The 
mixture was adjusted to a pH of 3-4 with acetic acid and was vacuum 
filtered. The recovered product was rinsed and dried to obtain 420 mg of 
raw product. The latter was dissolved in water containing sodium hydroxide 
and the solution was passed through an ion exchange resin column. Elution 
was with water and then with water containing 10% of isopropanol and the 
fractions with the product were combined and the solvents were evaporated 
under reduced pressure. The aqueous residue was acidified with acetic acid 
to obtain 115 mg of 
7--(2-amino-4-thiazolyl)-acetamido!-3-{(2-furyl)-carbonyl}thiomethyl!-ce 
ph-3-eme-4-carboxylic acid. 
Analysis: C.sub.18 H.sub.16 O.sub.6 N.sub.4 S.sub.3 : Calculated: %C, 45.0; 
%H, 3.4; %N, 11.7; %S, 20.0. Found: %C, 44.5; %H, 3.5; %N, 12.2; %S, 20.0. 
EXAMPLE 6 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-propionylthiomethyl-ceph-3-eme-4-ca 
rboxylic acid 
128 ml of ethanol were added to a mixture of 4.6 ml of propionyl chloride 
in a solution of 8 g of sodium sulfhydrate in 20 ml of water and the 
mixture was stirred for 30 minutes and was vacuum filtered. The filtrate 
was evaporated to dryness and the residue was taken up in ethanol. The 
solution was evaporated again and the residue was again taken up in 
ethanol. The mixture was vacuum filtered and the filtrate was evaporated 
to dryness. The residue was added to ether and vacuum filtered to obtain 
4.5 g of sodium thiopropionate. 
A mixture of 1.23 g of 
7--2-(2-amino-4-thiazolyl)acetamido!-3-acetoxymethyl-ceph-3-eme-4-carboxy 
lic acid, 12ml of water, 252 mg of sodium bicarbonate and 516 mg of sodium 
thiopropionate was heated at 60.degree. C for 4 hours and was then cooled. 
0.2 g of carbon black were added to the mixture which was then vacuum 
filtered. The product was acidified, washed and dried to obtain 0.88 g of 
raw product. 0.44 g of the raw product in 8.6 ml of methylene chloride and 
0.2 ml of triethylamine was stirred and then was vacuum filtered. The 
insolubles were re-extracted and vacuum filtered again. The combined 
filtrates were evaporated to dryness and the residue was dissolved in 
water. 0.2 ml of acetic acid were added to the solution and the mixture 
was vacuum filtered. The recovered product was washed and dried to obtain 
0.232 g of purified 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-propionylthiomethyl-cep-3-eme-4-ca 
rboxylic acid. 
Analysis: C.sub.16 H.sub.18 O.sub.5 N.sub.4 S.sub.3 : Calculated: %C, 
43.42; %H, 4.1; %N, 12.66; %S, 21.73. Found: %C, 43.3; %H, 4.3; %N, 12.2; 
%S, 20.8. 
EXAMPLE 7 
An injectable solution was prepared from 500 mg of 
7-2-(2-amino-4-thiazolyl)-acetamido!-3-acetylthiomethyl-ceph-3-eme-4-carb 
oxylic acid and sufficient sterile aqueous excipient for a total volume of 
5 ml. Gelules were prepared with 250 mg of the same product and sufficient 
excipient for a gelule with a final weight of 400 mg. 
PHARMACOLOGICAL DATA 
A. In Vitro Activity -- Dilution Method in Liquid Media 
A series of tubes were prepared with the same quantity of sterile nutrive 
media and increasing quantities of the test product was distributed to 
each tube. Then, each tube was seeded with a bacterial strain and after 
incubation at 37.degree. C for 24 or 48 hours, the inhibition of the 
growth was determined by transillumination which permitted determination 
of the minimum inhibitory concentrations (M.I.C.) in .mu.g/ml. The results 
are reported in Tables I to IV. 
TABLE I 
______________________________________ 
PRODUCT - EXAMPLE 2 
MIC in .mu.g/ml 
Bacterial Strain 24 H 48 H 
______________________________________ 
Staphylococcus Oxford UC 1061 
Penicillin sensible 0.5 0.5 
Staphylococcus aureus UC 1128 
Penicillin resistant 1 1 
Streptococcus hemolyticus (in bacto 
Todd Hewitt Broth pH = 7.8) 
0.05 0.1 
Streptococcus faecalis n.degree. 5 432 
5 >40 
Bacillus subtilis 0.1 0.5 
Escherichia Coli UC 1 020 
2 5 
Escherichia Coli UC 1 261 
1 1 
Klebsiella pneumoniae 52 145 
0.5 0.5 
Proteus mirabilis (indol) 
1 1 
Staphylococcus aureus 54 146 
0.6 0.6 
Escherichia Coli (T) 0 26 B 6 
3 3 
______________________________________ 
TABLE II 
______________________________________ 
PRODUCT - EXAMPLE 4 
MIC in .mu.g/cm.sup.3 
Bacterial strain 24 H 48 H 
______________________________________ 
Staphylococcus aureus UC 1 061 
Penicillin sensible 0.2 0.5 
Staphylococcus aureus UC 1 128 
Penicillin resistant 1 1 
Staphylococcus aureus n.degree. 54 146 
0.5 1 
Streptococcus pyogenes A 561 
0.1 0.2 
Streptococcus faecalis 5 432 
10 >40 
Bacillus subtilis 0.1 0.1 
Escherichia Coli UC 1 020 
2 2 
Escherichia Coli UC 1 261 
1 1 
Escherichia Coli T 026 B 6 
2 3 
Escherichia Coli R 55 123 D 
5 10 
Klebsiella Pneumoniae 52 145 
0.1 0.2 
Proteus mirabilis (indol) A 235 
0.6 0.6 
Salmonella Typhimurium 420 
3 3 
______________________________________ 
TABLE III 
______________________________________ 
PRODUCT - EXAMPLE 5 
MIC in .mu.g/cm.sup.3 
Bacterial Strain 24 H 48 H 
______________________________________ 
Staphylococcus aureus UC 1 061 
Penicillin sensible 0.5 1 
Staphylococcus aureus UC 1 128 
Penicillin resistant 1 2 
Staphylococcus aureus n.degree. 54 146 
1 1 
Streptococcus pyogenes A 561 
0.1 0.1 
Streptococcus faecalis 5 432 
5 40 
Bacillus subtilis ATCC 6 633 
0.2 0.2 
Staphylococcus aureus ATCC 6 538 
0.5 1 
Escherichia Coli ST UC 1 020 
10 10 
Escherichia Coli RT UC 1 261 
2 2 
Escherichia Coli T 026B6 
10 10 
Escherichia Coli RG R 55 123 D 
10 10 
Klebsiella pneumoniae Exp. 52 145 
0.4 1 
Proteus mirabilis (Indol) A 235 
2 2 
Salmenella Typhimurium 10 
10 
______________________________________ 
TABLE IV 
______________________________________ 
PRODUCT - EXAMPLE 6 
MIC in .mu.g/cm.sup.3 
Bacterial Strain 24 H 48 H 
______________________________________ 
Staphylococcus aureus UC 1 061 
Penicillin sensible 0.6 0.6 
Staphylococcus aureus UC 1 128 
Penicillin resistant 1 1 
Staphylococcus aureus 54 146 
1 2 
Streptococcus pyogenes A 561 
0.2 0.2 
Streptococcus faecalis 99 F 74 
20 40 
Bacillus subtilis ATCC 6 633 
0.1 0.1 
Staphylococcus aureus ATCC 6 538 
0.2 0.4 
Escherichia Coli ST UC 1 020 
10 10 
Escherichia Coli RT UC 1 261 
10 10 
Escherichia Coli T 026B6 
40 40 
Klebsiella pneumoniae 52 145 
3 5 
Proteus mirabilis (Indol ) A 235 
10 20 
Salmonella Typhimurium 420 
5 5 
Serratia RG 2 532 10 10 
______________________________________ 
B. In Vivo Activity -- Experimental Staphylococcia 
The antibacterial activity of the compound of Example 2 was determined 
against experimental staphylococcia in mice. Lots of 10 male mice weighing 
about 21.5 g were infected by intraperitoneal injection of 0.5 ml of a 24 
hour culture of staphylococcus aureus 54,146 in Pasteur nutritive broth 
diluted by 1/5 with distilled water. The test product was administered 
subcutaneously 1 hour, 5 hours and 24 hours after the infection in 
different doses. The number of dead mice was determined during 8 days and 
the results are reported in Table V. 
TABLE V 
______________________________________ 
Dose ad- Mice 
ministered living 
in each Mortality after after 
injection 
9 h 45 22 h 30 25 h 30 
29 h 4 d 5 d 8th day 
______________________________________ 
0.1 mg 1 8 1 0/10 
0.25 mg 1 1 1 1 6/10 
0.5 mg 1 9/10 
0.75 mg 10/10 
1 mg 10/10 
______________________________________ 
C. In Vivo Activity -- Experimental Infection of Proteus Mirabilis 
The product of Example 2 was studied for its effect on an experimental 
infection of Proteus mirabilis in mice. Groups of 10 male mice weighing 
about 22.5 g were infected by an intraperitoneal injection of 0.5 ml of a 
24 hour culture in Pasteur nutritive media of Proteus mirabilis No. A 235 
diluted by 1/4 with distilled water. 
The mice received a subcutaneous injection 1 hour, 5 hours and 24 hours 
after the infection of a quantity of the product of Example 2 and the 
mortality was noted during 8 days. The results are in Table VI. 
TABLE VI 
______________________________________ 
Dose administered 
Mortality after 
Mice living on 
in each injecton 
21 h 15 8th day 
______________________________________ 
Controls 10 0/10 
0.1 mg 2 8/10 
0.25 mg 0 10/10 
0.5 mg 0 10/10 
0.75 mg 0 10/10 
______________________________________ 
D. Experimental Infection with Escherichia Coli (T) 026B6 
Again, the activity of the product of Example 2 was tested against 
experimental infection of Escherichia Coli in mice. Groups of 10 male mice 
weighing about 23 g were infected with an intraperitoneal injection of 0.5 
ml of a 24 hour culture in Pasteur nutritive media of Escherichia Coli (T) 
026B6 diluted by 1/6 with distilled water. The test product was 
administered subcutaneously one hour, five hours and 24 hours after the 
infection and the mortality was determined during 8 days. The results are 
reported in Table VII. 
TABLE VII 
______________________________________ 
Dose Admin- 
instered Mice 
in each 21 h 24 h 27 h 30 h 45 h Living 
injection 
15 15 45 45 30 70 h on 8th day 
______________________________________ 
Control 10 0/10 
0.5 mg 6 1 1 1 1 0/10 
1 mg 1 9/10 
2 mg 10/10 
3 mg 10/10 
______________________________________ 
The results of Tables I to VII clearly show the antibacterial activity of 
the tested compounds. 
Various modifications of the products and methods of the invention may be 
made without departing from the spirit or scope thereof and it should be 
understood that the invention is to be limited only as defined in the 
appended claims