FORM 2
THE PATENT ACT 1970 (39 of 1970)
& The Patents Rules, 2003
PROVISIONAL SPECIFICATION
(See section 10 and rule 13) i. TITLE OF THE INVENTION: "Synergistic parenteral compositions of Cephalosporins and Beta-Lactamase
inhibitors"
2. APPLICANT (S)
(a) NAME: LYKA LABS LIMITED
(b) NATIONALITY: an Indian Company incorporated under the
Indian Companies ACT, 1956
(c) ADDRESS: 101, Shivshakti Industrial Estate, Andheri Kurla Road,
Andheri (East), Mumbai - 400 059.
3. PREAMBLE TO THE DESCRIPTION
The following specification particularly describes the invention.

Technical field of the invention:
The present invention relates to synergistic pharmaceutical parenteral compositions having broad spectrum antibacterial activity comprising combinations of Cephalosporins
and Beta-Lactamase inhibitors. More particularly the invention relates to parenteral compositions wherein the Cephalosporins are Cefepime, Ceftazidime and Cefoperazone and the Beta-Lactamase inhibitors are Sulbactam and Tazobactam.
Background and Prior Art
Treatment and control of infections remain a major goal in day to day practice as this being more important in hospital environment where patients are admitted with serious ailments and also suffering from many chronic disease like diabetes, cancer etc. In hospital practice mixed bacterial infections are very common. In most instances resistant staphylococci and other enterobacteriaceae are commonly found. To add to the problems, treatment with cytotoxic or immunosuppressant drugs make patients more vulnerable to infections with bacteria which are more often resistant to routinely used antibiotics.
Chemical modification of 7-aminocephalosporanic acid molecule led to discovery of several oral and parenteral Cephalosporins. The third generation Cephalosporins such as Ceftriaxone, Cefotaxime and Ceftazidime were introduced into clinical practice in 80's and are considered as efficacious and fairly safe antibiotics in the management of many serious infections. Inspite of these discoveries antibiotic resistant bacteria remains most dreaded scenario in hospital practice. Such infection led to multi-organ failure and death. One of the approaches was to combine two antibiotics, like a cephalosporin and an aminoglycoside. Besides stability, when mixed together these combinations may lead to nephrotoxicity.
Beta-Lactamase production is one of the most important mechanism of antibiotic resistance in bacteria. The large number of Beta-lactamases produced by gram positive and gram negative bacteria are either plasmid or chromosomally mediated beta lactamases. The recent appearance of extended spectrum Beta-Lactamases (ESBLS), are capable of conferring resistance to the third generation Cephalosporins. Recently point
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mutations in TEM-1, TEM-2 and SHV-1 enzymes have resulted in more extended spectrum beta lactamases (ESBL), which are produced by E-coli, Klebsiella sp., Pseudomonas.
Cefepime is a bactericidal Cephalosporin belonging to fourth generation. It has broad spectrum in-vitro activity against wide range of gram positive and gram negative bacteria. Cefepime has low affinity for chromosomally encoded beta lactamases. Cefepime is highly resistant to hydrolysis by most beta lactamases and exhibits rapid penetration into gram negative bacterial cells. Within bacterial cells, Cefepime targets the penicillin binding proteins (PBP). Most strains of enterococci eg. Enterococcus faecalis and methicillin resistant staphylococci are resistant to Cefepime. Cefepime is not effective against ESBL organisms.
Ceftazidime is a semi-synthetic third generation Cephalosporin antibiotic. Its activity includes coverage against gram positive and gram negative bacteria and expanded spectrum of activity against gram negative bacteria. It is more active against Pseudomonas than other. Its activity is less against anaerobes and gram positive bacteria. Ceftazidime generally is stable against hydrolysis by B-lactamase classified as Richmond Sykes types I, II, III (TEM Type) IV and V, most PSE types. Resistance strains of Enterobacter and Pseudomonas have developed during therapy. Ceftazidime resistant strains of Klebsiella pneumoniae have been reported and these strains have been involved in nosocomial outbreaks in hospitals. Resistance in these strains results from acquisition of plasmid mediated extended spectrum B-lactamases (ESBL).
Cefoperazone is classified as a third generation Cephalosporin based on its spectrum of activity. Cefoperazone has an extended spectrum of activity against gram negative bacteria compared to first and second generation Cephalaosporins. However it is less active in vitro against susceptible staphylococci than a first generation Cephalosproin. Cefoperazone is more susceptible to inactivation by B-lactamases than are Cefotaxime, Ceftizoxine and Cefuroxime. Cefoperazone is generally resistant to inactiviation by B-lactamases that act principally as Cephalosporinases and is generally not hydrolyzed by enzymes produced by Pseudomonas aeruginosa. Cefoperazone is more active against Pseudomonas aeruginosa than many other available Cephalosporins and may be
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considered as drug of choice for the treatment of infection caused by pseudomonas aeruginosa.
Tazobactam Sodium has little clinically relevant in-vitro activity against bacteria due to its reduced affinity to penicillin binding proteins. Tazobactam is a B-lactamase inhibitor of the Richmond Sykes Class III penicillinase and Cephalo-sporinases. It varies in its ability to inhibit class II and IV penicillinases. Tazobactam does not induce chromosomally mediated B-lactamases. It is poor inducer of B-lactamases of gram -positive and gram negative organisms. It inhibits wide range of B-lactamases including the group 2 penicillinase from Staph- aureaus, the broad spectrum TEM and SHV -I B-lactamases.
Sulbactam has a much greater affinity for B-lactamases and the it quickly forms an enzyme inhibitor complex which targets B-lactamases. Sulbactam inactiviates both plasmid and chromosome medicated B-lactamases. In-vitro studies indicate that Sulbactam generally inhibits staphylococcal B-lactamases and B-lactamases classified as Richmond sykes types II, III (TEM type, HSVI) IV, V (PSE and OXA types). Sulbactam Sodium has minimal activity against bacteria when used alone.
US6900184 of Cohen Jonathan et al discloses pharmaceutical compositions of piperacillin with tazobactam in the presence of a buffer, preferably citrate, a particulate formation inhibitor, preferably EDTA optionally an aminoglycoside which when frozen and thawed or lyophilized and reconstituted forms a solution which has decreased particulate formation and method of treating bacterial infections in mammals.
US4956179 of Bamberg Peter et al discloses antibacterial synergistic compositions comprising combination of penicillin derivatives and cephalosporin derivatives and methods for the preparation of such compositions and a method for the treatment of infectious diseases.
WO0202095 of Batts Donald et al discloses a composition having antibacterial activity comprising a mixture of an oxazolidinone compound, sulbactam, and ampicillin active agents, demonstrating activity against resistant strains of bacteria and methods for using
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an oxazolidinone compound, sulbactam, and ampicillin to treat a bacterial infection are also described.
Object of the invention:
The main object of the invention is to provide synergistic pharmaceutical parenteral compositions having broad spectrum antibacterial activity comprising combinations of Cephalosporins and P-Lactamase inhibitors which ameliorate the resistance problems associated with increased and repeated use of P-Lactam antibiotics.
Summary of the invention:
The present invention discloses synergistic pharmaceutical parenteral compositions having broad spectrum antibacterial activity comprising combinations of Cephalosporins and P-Lactamase inhibitors. The invention further discloses parenteral compositions wherein the Cephalosporins are selected from the group of Cefepime, Ceftazidime and Cefoperazone and the P-Lactamase inhibitors are selected from the group of Sulbactam and Tazobactam.
According to one aspect, the present invention discloses synergistic injectable formulations wherein the said Cephalosporin is formulated with B-lactamase inhibitor Tazobactam in the ratio of 8:1.
As per another aspect, the present invention discloses synergistic injectable formulations wherein the said Cephalosporin is formulated with B-lactamase inhibitor Sulbactam in the ratio of 2:1.
Description of the invention:
The present invention describes synergistic pharmaceutical parenteral compositions having broad spectrum antibacterial activity comprising combinations of Cephalosporins and Beta-Lactamase inhibitors. The invention particularly describes parenteral compositions wherein the Cephalosporins are Cefepime, Ceftazidime and Cefoperazone and the Beta-Lactamase inhibitors are Sulbactam and Tazobactam.
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Increased and repeated use of Beta-Lactam antibiotics leads to them becoming ineffective or less effective, principally as a result of the onset and worldwide spread of enzymatic resistance via Beta-Lactamase production by bacteria.
To counter this resistance problem, the inventors of the present invention have developed parenteral combinations of Beta-Lactamase inhibitor and penicillin such as a Cephalosporin. The rationale for such combined therapy is based on a synergistic effect of the two molecules, the activator that is Tazobactam destroys the Beta-Lactamase activity whereby the Cephalosporins are protected from inactivation.
According to one aspect of the present invention, the Cephalosporins such as Cefepime, Ceftazidime and Cefoperazone are formulated with the Beta-lactamase inhibitor Tazobactam in the ratio of 8:1.
Cefepime with Tazobactam is a combination formulated to reduce the drug-resistance of bacteria and maintain effectiveness of Cefepime. This combination should be used only to treat or prevent infection that are proven or strongly suspected to be caused by bacteria. Cefepime with Tazobactam is active against Enterobacteriaceae that are resistant to other Cephalosporins via induction of type -I beta- lactamases. It remains susceptible to many bacteria expressing extended spectrum plasmid-mediated Beta-lactamases such as TEM-3 and TEM-10 against the fastidious gram negative bacteria H-influenzae, N. gonorrhoeae and N-meningitidis. Combination of Cefepime and Tazobactam has greater or comparable in vitro activity than Cefotaxime. For Pseudomonas aeruginosa, Cefepime and Tazobactam combination has comparable activity to Ceftazidime. Combination of Cefepime and Tazobactam has higher activity than Ceftazidime and Cefotaxime for Streptococci and Methicillin sensitive staphylococci.
Combination of Tazobactam with Cefepime also has some advantage over other beta-lactam/ beta-lactamase inhibitor combinations such as co-amoxiclav, ticarcillin/clavulanic acid and ampicillin / sulbactam. Cefepime is easier to protect against TEM-beta lactamases. This is because of the lower affinity of Cefepime for these enzymes. Tazobactam appears to be a weaker enzyme inducer than other beta-lactamase inhibitor. Cefepime with Tazobactam combination include coverage for mixed gram-negative,
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anaerobic infections (including intra-abdominal, diabetic skin or soft tissue infections and nosocomial respiratory tract infections and empiric broad spectrum therapy for Polymicrobial infections). Their activity against Enterobacter, Pseudomonas and other
gram negative organisms provide enhanced coverage over ampicillin - sulbactam or amoxicillin- clavulanate for nosocomial infections.
Combination of Cefepime and Tazobactam is used for the treatment of:-
i. Empiric Therapy for febrile neutropenic patients.
ii. Uncomplicated and complicated urinary tract infections.
iii. Uncomplicated skin and skin structure infections.
iv. Complicated intra - abdominal infections.
Dosage: Cefepime and Tazobactam combinations are usually administered in a dose of 0.5 to lg every 12 hours, intravenously. In severe infections 2g may be administered intravenously every 12 hours.
Ceftazidime is more bactericidal and has similar plasma half-life of 1 hour like Tazobactam. The combination of Ceftazidime and Tazobactam is effective in the management of hospital acquired infection like nesocomial pneumonia, uncomplicated and complicated skin and skin structure infections. To avoid drug resistance of Staphylococci, Pseudomonas, Klebsiella and ESBL producing bacteria it is justified to combine Ceftazidime with Tazobactam. The combination may be used as empiric therapy for febrile neutropanic patients, infections in immunocompromised patients and empiric therapy of infections in patients with multiple organ failure.
Cefoperazone and Tazobactam is a combination of third generation Cephalosporin. Cefoperazone with Tazobactam a synthetic B-lactamase inhibitor. Tazobactam is B-lactamase inhibitor and it is synergistic with Cefoperazone. Tazobactam inhibits all beta-lactamases inhibited by other beta-lactamases inhibitors like clavulanic acid. But in addition it is also active against chromosomally mediated enzyme. Tazobactam is also a weaker enzyme inducer than other beta-lactamase inhibitors. ESBLS are inhibited by Tazobactam. Therefore combination of Cefoperazone with Tazobactam is considered
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essential for the treatment of multiply resistant microorganisms, mixed pseudomonas and gram positive infections, mixed aerobic and anaerobic infections.
Tazobactam with antibiotics in 1:8 ratio is effective and achieves therapeutic results. Ceftazidime and Cefepime when combined with Tazobactam will provide coverage against extended spectrum (^-Lactamases producing (ESBL) bacteria. Tazobactam not only enhances the coverage but protects the antibiotics from destruction by [3-Lactamase enzymes.
According to another aspect of the present invention, the Cephalosporins such as Cefepime, Ceftazidime and Cefoperazone are formulated with the B-lactamase inhibitor Sulbactam in the ratio of 2:1.
The combined use of Sulbactam with certain penicillins and Cephalosporins expands the spectrum of activity against many strains of B-lactamase producing bacteria. Results of invitro studies indicate that 2:1 ratio of antibiotic to Sulbactam results in optimal B-lactamase inhibition and anti-bacterial activity.
The combination of Sulbactam with Cefepime has advantage over other beta-lactam/ beta-lactamase inhibitor combinations such as co-amoxiclav, ticarcillin /clavulanic acid and ampicillin / sulbactam. Cefepime is easier to protect against TEM-beta lactamases. This is because of the lower affinity of Cefepime for these enzymes. Besides Sulbactam has an affinity for and binds to same penicillin binding proteins and this binding may also contribute to the synergistic bactericidal effect.
Cefepime with Sulbactam combination include coverage for mixed gram-positive, anaerobic infections (including intra-abdominal, diabetic skin or soft tissue infections and nosocomial respiratory tract infections and empiric broad spectrum therapy for Polymicrobial infections.) Their activity against staphylococci and other gram negative organisms provide enhanced coverage over ampicillin - sulbactam or anoxicillin-clavulanate for nosocomial infections.
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Combination of Cefepime and Sulbactam is used for the treatment of:-
i. Postoperative Infections
ii. Uncomplicated and complicated urinary tract infections.
iii. Respiratory tract infection including nosocomial pneumonia
iv. Complicated intra - abdominal infections.
Dosage: Combination of Cefepime and Sulbactam is administered in a dose of 0.5 to lg every 12 hours, intravenously. In severe infections 2g may be administered intravenously every 12 hours.
The processes for preparation of the said parenteral compositions are described followed by the non limiting examples.
Process A: Cefepime and Tazobactam for injection
Cefepime and Tazobactam injection is prepared by blending the two ingredients by
geometric mixing.
Blending
Stepl: Weigh and transfer Tazobactam Sodium and equal amount of Cefepime
Hydrochloride in a pre-sterilized cone blender. Run blender at appropriate speed for
specified period of time.
Step2: Weigh and transfer remaining amount of Cefepime Hydrochloride geometrically in
the above mixture (Stepl) and run the blender for specified period of time.
Process B: Ceftazidime and Tazobactam for injection
Ceftazidime and Tazobactam injection is prepared by blending the two ingredients by
geometric mixing.
Blending
Stepl: Weigh and transfer Tazobactam Sodium and equal amount of Ceftazidime
Pentahydrate in a pre-sterilized cone blender. Run blender at appropriate speed for
specified period of time.
Step2: Weigh and transfer remaining amount of Ceftazidime Pentahydrate geometrically
in the above mixture (Step 1) and run the blender for specified period of time.
Process C: Cefoperazone and Tazobactam for injection
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Cefoperazone and Tazobactam injection is prepared by blending the two ingredients by
geometric mixing.
Blending
Stepl: Weigh and transfer Tazobactam Sodium and equal amount of Cefoperazone
Sodium in a pre-sterilized cone blender. Run blender at appropriate speed for specified
period of time.
Step2: Weigh and transfer remaining amount of Cefoperazone Sodium geometrically in
the above mixture (Step 1) and run the blender for specified period of time.
Process D: Cefepime and Sulbactam for injection
Cefepime and Sulbactam injection is prepared by blending the two ingredients by
geometric mixing.
Blending
Stepl: Weigh and transfer Sulbactam Sodium and equal amount of Cefepime
Hydrochloride in a pre-sterilized cone blender. Run blender at appropriate speed for
specified period of time.
Step2: Weigh and transfer remaining amount of Cefepime Hydrochloride geometrically in
the above mixture (Stepl) and run the blender for specified period of time.
Process E: Ceftazidime and Sulbactam for injection
Ceftazidime and Sulbactam injection is prepared by blending the two ingredients by
geometric mixing.
Blending
Stepl: Weigh and transfer Sulbactam Sodium and equal amount of Ceftazidime
Pentahydrate in a pre-sterilized cone blender. Run blender at appropriate speed for
specified period of time.
Step2: Weigh and transfer remaining amount of Ceftazidime Pentahydrate geometrically
in the above mixture (Step 1) and run the blender for specified period of time.
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The present invention is more specifically explained by following examples. However, it should be understood that the scope of the present invention is not limited by the examples in any manner. It will be appreciated by any person skilled in this art that the present invention includes the following examples and further can be modified and
altered within the technical concept of the present invention.
Examples
Example 1

Ingredients Quantity
Cefepime Hydrochloride USP equivalent to Cefepime 500mg
Sulbactam Sodium USP equivalent to Sulbactam 250mg
Example 2

Ingredients Quantity
Cefepime Hydrochloride USP equivalent to Cefepime lOOOmg
Sulbactam Sodium USP equivalent to Sulbactam 500mg
Example 3

Ingredients Quantity
Cefepime Hydrochloride USP equivalent to Cefepime 500mg
Tazobactam Sodium USP equivalent to Tazobactam 62.5mg
Example 4

Ingredients Quantity
Cefepime Hydrochloride USP equivalent to Cefepime lOOOmg
Tazobactam Sodium USP equivalent to Tazobactam 125mg
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Example 5

Ingredients Quantity
Ceftazidime Pentahydrate IP equivalent to Ceftazidime lOOOmg
Tazobactam Sodium equivalent to Tazobactam 125mg
Example 6

Ingredients Quantity
Ceftazidime Pentahydrate IP equivalent to Ceftazidime 500mg
Tazobactam Sodium equivalent to Tazobactam 62.5mg
Example 7

Ingredients Quantity
Cefoperazone Sodium USP equivalent to Cefoperazone lOOOmg
Tazobactam Sodium equivalent to Tazobactam 125mg
Example 8

Ingredients Quantity
Cefoperazone Sodium USP equivalent to Cefoperazone 500mg
Tazobactam Sodium equivalent to Tazobactam 62.5mg
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It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative examples and that the present invention may be embodied in other specific forms without departing from the essential attributes thereof, and it is therefore desired that the present embodiments and examples be considered in all respects
as illustrative and not restrictive, reference being made to the appended claims, rather than to the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Dated this 22nd day of November 2006
Dr. Gopakumar G. Nair Agent for the Applicant
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