MXPA06008605A - Compositions substantially free of galactomannan containing piperacillin and tazobactam - Google Patents

Abstract

The invention pertains to pharmaceutical compositions of Zosyn®having substantially free or reduced levels of galactomannan and processes to prepare said pharmaceutical compositions.

Description

COMPOSITIONS CONTAINING PIPERACILLIN AND TAZOBACTAM FIELD OF THE INVENTION The invention relates to pharmaceutical compositions of Zosyn® substantially free of galactomannan.

BACKGROUND OF THE INVENTION Zosyn® is an antibiotic product marketed in the United States and Tazocin brand in many foreign countries containing piperacillin sodium and tazobactam sodium. The product is described in the US patent No. 4,562,073. U.S. Patent Nos. 4,477,452 and 4,534,977 describe a lyophilized form of piperacillin.

Zosyn® is an antibiotic that is used in the treatment of moderate to severe infections. In particular, Zosyn® is used in the treatment of moderate to severe infections caused by strains of beta-lactamase producing microorganisms susceptible to piperacillin / tazobactam, resistant to piperacillin under conditions such as nosocomial pneumonia due to Staphylococcus aureus; intra-abdominal infections, specifically appendicitis (complicated by rupture or access) and peritonitis due to Escherichia coli. Infections of the skin and the structure of the skin, including cellulitis, cutaneous access and ischemic / diabetic foot infections due to Staphylococcus aureus; and gynecological infections, specifically postpartum endometritis or pelvic inflammatory disease due to Escherichia coli. The seriousness of these infections highlights the need for an easily available and approachable treatment.

The drugs are formulated not only in emulsions, suspensions or solutions, but also as lyophilized preparations that are reconstituted before use. Advantageously, the lyophilized preparations are stable, can be stored and are easily reconstituted. Moreover, the lyophilized preparations can be kept sterile and essentially free of insoluble matter.

Zosyn® is available as a powder (lyophilized product) which is reconstituted by adding a compatible reconstitution diluent before intravenous administration. Zosyn® has been found to contain small amounts of galactomannan which is a carbohydrate polymer derived from fungal cell walls and formed in fermentation processes. The presence of galactomannan has been shown to interfere and deliver false positives in certain diagnostic assays for invasive aspergillosis (IA). Although present, galactomannan does not create an increased health risk to the patient.

The disadvantages of the presence of galactomannan in pharmaceutical compositions of Zocyn® are solved by the present invention.

BRIEF DESCRIPTION OF THE INVENTION Zosyn® is known to contain small amounts of galactomannan, a polymer of carbohydrates derived from the cell walls of fungi. However, although present, galactomannan does not create an increased risk to the patient's health.

Invasive aspergillosis (IA) is a fatal infection seen more frequently in immunocompromised patients. The presence of circulating galactomannan aspergillus antigen in the serum is an indicator of invasive aspergillosis (I A), a fatal fangal infection. Immunocompromised ponteros are often subject to prophylactic treatment with Zosyn® to prevent bacterial infections. The diagnosis of invasive aspergillosis in patients is often based on serological methods by detecting the presence of aspergillus galactomannan. The presence of very small amounts of galactomannan in Zosyn®, however, leads to false-positive test results as to when certain diagnostic kits are used. Removal of galactomannan from Zosyn® has the advantage of eliminating or diminishing false-positive results in potential IA diagnostic tests when such kits are used.

The present invention provides a new pharmaceutical composition of pre-blended piperacillin or piperacillin-tazobactam which avoids the presence of galactomannan and is useful for the treatment or control of bacterial infections by parenteral administration., the composition comprises effective amounts of (a) piperacillin or a pharmaceutically acceptable salt thereof (usually as piperacillin sodium), and (b) tazobactam or a pharmaceutically acceptable salt thereof (usually as tazobactam sodium). The pharmaceutical composition according to the invention can be (A) in the form of a powder that can be reconstituted by the addition of a compatible reconstitution diluent before parenteral administration, (B) in a ready-to-use form in parenteral administration. or (C) in a frozen form that can be thawed and is ready to use for parenteral administration. The composition of the invention is provided substantially free of galactomannan.

The invention further includes: A process for preparing a lyophilized pharmaceutical composition that is substantially free of galactomannan comprising the steps of: a) dissolving the piperacillin, and the tazobactam in an aqueous solvent forming a solution and adjusting the pH to about 6.5; b) filter the solution by means of a cut filter; c) reconnect a filtrate; d) cool the filtrate to a temperature below -35 ° C in a lyophilizer, e) evacuate the lyophilizer to a pressure of about 300 μm Hg (micrometers of mercury) (40 pass) and heat the lyophilizer to about +5 ° C; f) maintaining the temperature and pressure for a sufficient time to remove the water from the aqueous solvent forming a lyophilized solid; g) Dry the lyophilized solid at about + 45 ° C.

The invention also includes a process for the manufacture of a pharmaceutical composition in the form of a powder that can be reconstituted by the addition of a compatible reconstitution diluent before administration to a mammal or in the form of a frozen composition that can be thawed with a compatible diluent prior to administration to a mammal said process comprises freezing or freeze-drying a solution substantially free of galactomannan containing effective amounts of (a) piperacillin or a pharmaceutically acceptable salt thereof, (b) tazobactam or a pharmaceutically acceptable salt thereof in an aqueous vehicle.

DESCRIPTION OF THE PREFERRED MODALITIES The present inventive composition offers an advantage over other forms of piperacillin and piperacillin-tazobactam for administration. In particular, the invention provides a composition that is substantially free of galactomannan. Without the presence of galactomannan in the composition of Zosyn® there is a lack of interference and false positive results in the assay with antibody assays that are used for the determination of invasive aspergillosis. Critical to the removal or reduction of galactomannan is the use of an appropriate cut filter from about 3 kD mw to about 10 kD mw. The galactomannan is collected on the filter and the piperacillin or piperacillin-tazobactam passes through the filter and is collected in the collected filtrate. A molecular weight cutoff filter of about 3 kD is preferred. More preferably it is a cut filter of about 5kD.

The removal or reduction of galactomannan proceeds as follows: an aqueous solution of Zosyn® at about (10 mg / ml) is prepared. The solution is applied to series of micro centrifuged filter devices (Pall Life Sciences) and the filters are centrifuged at 10,000 X g. This procedure forces the solution through the ultrafiltration membrane. The solutes don separated by the membrane based on molecular weight. Low molecular weight materials, such as piperacillin and tazobactam, pass through the ultrafiltration membrane (filtrate) while materials with a molecule weight greater than the cutoff membrane are effectively retained by a filter (retentate). Galactomannan is known to have a high molecular weight of between 25,000 to 75,000; piperacillin and tazobactam have low molecular weights < 1000. When a solution of Zosyn® contains galactomannan is applied to a cut-off filter of molecular weights of 3000 mw and is rotated, the galactomannan is found in the re-stain (R). The filtrate (F) contains the components of piperacillin and tazobactam and the filtrate tests are negative for galactomannan. Similar results are found with a membrane of 5 kD. The results found using a 10-kD cut filter show that minor amounts of galactomannan are in the filtrate. Importantly, there is no loss in the strength of piperacillin and tazobactam in the filtrate when compared to the starting material. In typical experiments, where progress is followed by high pressure liquid chromatography (HPLC) the following results are obtained after ultrafiltration. 1. Zosyn® Recovery of tazobactam- 100.3% Recovery of Piperacillin Monohydrate- 99.0% 2. Piperacillin- 100.1% 3. Ampicillin- 99.8% This process is easily adapted to scale production for commercial operations using currently available ultrafiltration (UF) devices and available ultrafiltration membranes.

Galactomannan can be effectively removed from Zosyn® solutions by ultrafiltration. The work has shown that filtration through appropriate molecular weight cutting membranes separates the high molecular weight galactomannan from the components of the low molecular weight Zosyn®. A further removal of galactomannan and an increased recovery of piperacillin and tazobactam can be further carried out in commercial operations using diafiltration with membrane filters as a portion of the ultrafiltration with shear filter. The membrane filters in the diafiltration retain the galactomannan and allow the Zosyn® components to pass through and be collected in the filtrate. Galactomannan can also be removed from 6-aminopenicillanic acid (6-APA) and ampicillin by the appropriate membrane filter.

EXPERIMENTAL PROTOCOL TITLE: Evaluation of Zosyn®, active pharmaceutical ingredient (API) and other antibiotics regarding the presence of galactomannan using the Aspergilius EIA method of BIO-RAD Platelia® 1. Purpose The purpose of this protocol is to describe the experimental design for the evaluation of different batches of Zosyn®, API and other antibiotics regarding the presence of the galactomannan antigen using the Aspergilius EIA kit of BIO-RAD Platelia®. 2. Materials and Equipment 2.1 Samples and reagents 1. Samples Zosyn® 2.250 g / bottle Zosyn® 4.5 g / bottle Tazoxil 4.5 g (generic Zosyn®) from Brazil Tazac 4.5 g (generic Zosyn®) from India piperacillin Tazobactam 4.5 g (Zosyn® generic) Richet (Argentina) Other products that are included in this protocol to evaluate their response in the Aspergilus EIA diagnostic kit of BIO-RAD Platelia®. 2. Aspergilius EIA Platelia® (BIO-RAD, Redmond, WA), No. 62793 (96 Test Kit) or No. 62794 (480 Test Kit) 2. 2 Equipment 1. Microplate reader: Dymex MRX ELISA microplate reader 2. Ultrawash ll automatic washer / Vacuum cleaner, Dynex 3. Biosecurity cabinet 4. Boiling water bath 5. Incubator 6. Vortex shaker 7. Sterile tubes, sterile gloves and sterile pipette tips. 8. Micropipette 3. Environmental control Preparation of reagents, sample and dilutions of samples are made under aseptic conditions in a biosafety cabinet. 4. Zosyn® assay site The experiments are conducted at the Chemical Processes and Biochemistry Development Laboratory, Wyeth Research, Pearl River, NY.

. Main Test and Procedure 5.1 Main Test The EIA Aspergilus Platelia® is a one-step immunoenzymatic sandwich microplate assay used for the detection of galactomannan in human serum. A rat single-channel EBA-2 antibody is used to capture the antigen, which is then detected using an antibody conjugated with peroxidase. The absorption capacity value of the sample is compared to the absorption capacity value of the "cut" control, thus determining the relative concentration / index of galactomannan. . 2 Procedure It refers to the user manual of the Aspergilus EIA kit of BIO-RAD Platelia® for the preparation of the reagent, stage-by-stage test procedure and safety instructions on the handling of reagents in the wheels.

Sample Preparation: A USP grade (US Pharmacopoeia) or any other suitable diluents is reconstituted in water for injection (WFI) and dilutions are made at desired concentrations.

The dilution of the sample can be changed based on the results of the experimental procedures. 6. Experimental Design 6.1 Product Evaluation 1. Evaluation of Zosyn® Vials of Zosyn® analysis at desired concentrations in water for injection (WFI) / phosphate buffer (PBS) or other appropriate matrix. 2. Evaluation of active pharmaceutical ingredients (API) Analysis bottles of piperacillin and tazobactam, and any other intermediate available in water for injection (WFI) / phosphate buffer (PBS). 3. Generic Zosyn® and / or other antibiotics Analysis of other available generic Zosyn® / antibiotics at desired concentrations in WFI / PBS. 6. 2 Filtration studies Zosyn® 1. The reconstituted samples of the filter using appropriate molecular weight cutoff filters and the filtration test at appropriate concentrations. Evaluate other studies on filtration capabilities as appropriate. 3. Acceptance Criterion Cutting control: The optical density (OD) 450 of each (2) Serum well control cut should be between 0.3 and 0.8. Each individual value should comply with the specification. The Average of Court. The control is averaged between two well readings, (see BIORAD kit instructions). Positive Control: The positive control serum index should be greater than 2. Negative control: The negative control serum index should be less than 0.4. Failure in any of the controls to meet the criteria renders the test invalid. To determine the index for the experimental samples, divide the absorption capacity (OD450) of the test sample by the average of the cutoff control. An index greater than 0.5 is considered a positive result. An index less than 0.5 is considered a negative result. 4. REFERENCES Aspergilius EIA Platelial® Manual (BIO-RAD, Redmond, WA). 1 = Positive Control of OD (R5) > 2 1 = Negative control of OD (R3) < 0.4 OD of Average Cutting Control STRENGTH OF ZOSYN® (PIPERACILLINE / TAZOBACTAM) AND IDENTIFICATION IN AQUEOUS SAMPLES BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY 1. METHOD DESIGN A portion of the Zosyn® sample is dissolved and diluted with dilution solvent and then chromatographed on a reversed phase column (USP 23 NF18, Vol.25, p.7497, Supp.6, p.3722). The forces of piperacillin and tazobactam are determined by comparing the respective peak responses in the chromatogram of the sample preparation for those between standard chromatograms obtained concomitantly. Piperacillin and tazobactam are identified by comparing the retention times of the respective peaks in the chromatogram of the sample preparation with those of the respective peaks in the standard preparation chromatograms. The method that reports the limit for piperacillin is 0.16 μg / mL for the injected solution. The method that reports the limit for tazobactam is 0.077 μg / mL for the injected solution. 2. SPECIAL EQUIPMENT Chromatographic Column- Length about 25 cm, internal diameter about 4.6 mm, packed with Fenomenex Luna C18 (2), particles size 5 μm.

NOTE: Columns of lengths 150 mm to 300 mm can be used as long as the system's adaptability requirements are met. Pump - Constant flow pump capable of operating at pressures up to 5000 psi.

Detector - Ultraviolet photometric spectrum detector capable of operating at 220 nm with a sensitivity of about 1.0 units of full-scale absorption capacity. Injector - Any manual injector or auto injector capable of reproducible injections and maintain a sample tray temperature of 5o C. Integrator- Electronic integration is preferred. Recorder- Optional. A recording device that matches the output voltage of the detector operation. Membrane filter - Pore size 0.45 μ, nylon 66 membrane filters. Column temperature controller - able to maintain a column temperature of 30 ° C. 3. REAGENTS AND MATERIALS Methanol-grade HPLC. Monobasic Sodium Phosphate- (NaH2P04) reactive grade. 0.4 M tetrabutylammonium hydroxide - Reagent grade. Phosphoric acid- 85%, reactive grade. Water-Suitable for HPLC. A solution of monobasic sodium phosphate buffer at 0.2 M- weight 27.6 g of monobasic sodium phosphate and diluted in 1 L with water. 20% phosphoric acid solution - diluted 23.5 mL of 85% phosphoric acid in 100 mL of water and mixed. 2% phosphoric acid solution - diluted 2.4 mL 85% phosphoric acid in 100 mL with water and mixed. Dilution solvent - Mobile phase. Mobile phase - measured in 447 mL of water, add 100 mL of a buffer solution of sodium phosphate monobasic to 0.2 M, pipette 3.0 mL of tetrabutylammonium hydroxide and add 450 mL of methanol. Mix. Cool to room temperature. Adjust the pH of the solution to approximately 5.6 with the 20% phosphoric acid solution and then 5.50 ± 0.02 with the 2% phosphoric acid solution. Filter through a membrane filter with pore size 0.45 μm, if necessary. Degas if necessary. Standard reference piperacillin- of known strength (S). Standard reference of Tazobactam- of known strength (S). 4. PREPARATION OF THE EQUIPMENT 1. Set the wavelength of the detector at 220 nm and the sensitivity around 1.0 units of absorption capacity in full scale. (The setting of the sensitivity may vary depending on the device used). 2. Set the flow rate at 0.8 mL per minute (0.5 to 1.2 mL per acceptable minutes). 3. Set the temperature controller of the column at 30 ° C. 4. Set the temperature controller of the injector / autosampler at 5o C; . The mobile phase pump through the column until a stable baseline is obtained (usually about 15 X column volume).

. STANDARD PREPARATION 1. Accurate weight around 24 mg of Tazobactam as standard reference, and 20 mg of Piperacillin as standard reference in 2 separate 50 ml volumetric flasks. 2. Dissolve the standards with a few drops of methanol (sonicate if necessary) and dilute the Tazobactam to a volume with dilution solvent. This is the standard starting solution of Tazobactam. 3. A pipette of 5.0 ml of the standard starting solution of Tazobactam in the piperacillin bottle. Dilute the volume with dilution solvent and mix. This is the standard preparation of Piperacillin / Tazobactam. (Approximately 400/48 μg / mL, respectively). These are standard calculations for single point. 4. (This stage is only required when the vehicle / control samples are being tested). Pipette 2.0 mL of the standard Piperacillin / Tazobactam preparation (400/48 μg / mL) into a 100 mL volumetric flask and dilute to volume with the dilution solvent. Pipette 2.0 mL of this solution each into volumetric flasks of 100 and 25 mL and dilute to volume with the dilution solvent. These are the standard limit preparations for reporting Piperacillin and Tazobactam, respectively, (approximately 0.16 μg / mL Piperacillin for the first solution and 0.077 μg / mL Tazobactam for the second solution). For each of these preparations only the relevant concentration is used.

NOTE 1: The linearity for Piperacillin has been established between 100 to 500 μg / mL. The linearity for Tazobactam has been established between 10 to 100μg / mL. Standard weights proportionately smaller or larger may be taken, provided that any subsequent dilution is adjusted accordingly to provide the standard preparation concentrations within the linear range. If this is done, the appropriate adjustments should be made to the calculations.

NOTE 2: Other dilution systems are possible as long as the final dilutions and the injected concentrations are within the linear range. If this is done, the appropriate adjustments should be made to the calculations. 6. PREPARATION OF THE SAMPLE Based on the claimed concentrations of the sample, make necessary dilutions in the dilution solvent to obtain a concentration of sample solution close to the standard single point concentrations for Piperacillin and Tazobactam (approximately 400 and 48 μg / mL, respectively ). For the pre-measured sample of more or less 2 mL typical, quantitatively transfer the entire sample. Rinse the bottle, cover the bottle, and out of the neck of the bottle add the rinses to the dilution bottle.

If necessary, stir the sample bottle during rinsing to remove the entire sample. Dilute to volume with dilution solvent and mix well. Any subsequent dilutions should also be made in the solution solvent. Samples should be processed one at a time to minimize the time before being injected.

NOTE 1: Non-typical samples may require an alate preparation procedure. For example, the volume of the sample or concentration may require an aliquot to be taken.

NOTE 2: The 2:10 vehicle / control diluted samples for the typical 20 Ml sample for piperacillin. For Tazobactam further dilute the sample 2:10.

If the samples are pre-filled, the initial sample volume should be calculated using the density as follows: (mL) Sample volume = (g) sample mass / (g / mL) Density 7. ADEQUACY OF THE SYSTEM 1. After a stable baseline has been obtained, inject 10 μL of the piperacillin / Tazobactam standard preparation three times and obtain a chromatogram of the Piperacillin / Tazobactam reference standard. These injections are used for the suitability of the system and the calculations. 2. Calculate the capacity factor, k 'of Piperacillin. The capacity factor must be 3.5 or greater. If not, prepare a fresh mobile FACE or replace the column.

NOTE: The ta value (retention time of a non-retained peak) should be estimated by dividing 60% of the volume of the column by the flow rate in ml_ / minutes. For the specified Phenomenex column, the estimate of ta is 2.5 mlJ (flow rate in mL / minutes). 3. Calculate the column coefficient of the column, T, as directed in the USP. The column coefficient of the column should not be more than 1.5. If it is greater, repair the chromatographic system and / or replace the column. 4. Calculate the theoretical plates, N, as directed in the USP. The value of N must be greater than or equal to 3000. If it is less, decrease the flow velocity within the permitted range, replace the column and / or repair the chromatographic system.

. Calculate the RSD for the three replicate Piperacillin injections. The RSD should not be more than 2.0%. 8. PROCEDURE A. Strength 1. (This stage is required only when the vehicle / control samples are being tested). At some point during the test, inject 10μL of the dilution solvent to obtain a blank chromatogram. Inject 10 μL of the sample preparations and standard preparations of the reporting limit and obtain the responses in the retention time of the peak of interest. 2. Inject 10μL of the sample preparation and obtain the responses of the peaks of interest.

B. Identification. 1. Inject 10 μL of each of Piperacillin / Tazobactam and obtain the retention time of the respective peaks. 2. Inject 10μL of the sample preparation and obtain the retention time of the respective peaks. 9. CALCULATIONS A. Strength 1. Calculate the concentration of Piperacillin / Tazobactam of the standard preparation from the following equations: Piperacillin mg / ml = (Wr) (S) / (50) mg Tazobactam / ml = (Wr) (S) (V1) / (50) (V2) Where: Wr = weight of the respective reference standard, mg S = strength of the respective reference standard, decimal V1 = volume of the standard starting solution used to make the standard preparation, mL. 50 = Volume of the standard starting solution or standard preparation, mL V2 = Volume of the standard preparation, mL 2. For Piperacillin and Tazobactam: Calculate the forces from the equation: mg Piperacillin or Tazobactam / mL = ( Cs) (Rspl) (Dspl) / (Rstd) Where: Cs = concentration of the respective standard from 1 up, in mg / mL Rspl = Response for the sample priority Dspl = Dilution factor for sample preparation Rstd = Average response for the respective standard preparation B. Identification 1. Calculate the relative retention value (Rr) of the respective peak in the chromatogram of the sample preparation using the expression: Rr = Rt of the respective peak, from the chromatogram of the sample / Rt of the respective peak, from the standard chromatogram.

Rt = Retention time, minutes 2. Identity report as positive and: Rr is 1.0 ± 0.05, otherwise it is reported that the identity is negative.

. REPORT LIMIT The limit of the report for Piperacillin for this model is 0.16 μg / mL for the injected solution. This is 0.8 μg / mL for a vehicle / control sample of 2 mL diluted in 2 mL to 10 mL. The reporting limit for Tazobactam for this method is 0.077 μg / mL for the injected solution. This is 1.92 μg / mL for a 2 mL vehicle / control sample diluted between 2 mL to 10 mL and then between 2 mL and 10 mL again.

FILTRATION STUDY The Zosyn® (typical commercial sample) is dissolved in water at 10 mg / ml. Piperacillin is dissolved in saturated sodium bicarbonate at 100 mg / ml. Zosyn® and Piperacillin are diluted 10 to 1 mg / ml using USP water. Zosyn® (300 μl) at 10 and 1 mg / ml as well as Piperacillin are transferred to the nanosep spinning device with 10 kD or 3 kD molecular weight cut-off filters. The samples are placed in an eppendorf centrifuge and centrifuged for 10 minutes at 10,000 rpm. At the end of the centrifugation, the samples are collected in the penetration. The galactomannan retained at the top of the nanosep spinning device is resuspended with 300 μl of water for the assay. Typical results are shown in the following Examples 1-4. The optical density (OD) for galactomannan are shown for each example, as well as the determined index of experimental samples.

Results: negative CTL: 0.078, index = 0.14 CTL C-O: 0.534, 0.554, average optical density (OD) = 0.544 positive CTL: 2.009, index 3.69 EXAMPLE 1 Filter 10K (10Kd) * (R) is the retentate (retained in the filter) ** (F) in the filtering EXAMPLE 2 Filter 3K (3kD) * (R) is the retained (retained in the filter) ** (F) in the filtering EXAMPLE 3 Filter 10K (10kD) * (R) is the retentate (retained in the filter) ** (F) in the filtering EXAMPLE 4 3K filter (3 kD) (R) is the retentate (retained in the filter) * (F) in the filtrate EXAMPLE 5 The experimental activity consisted in: (1) formulating a bulk product of ZOSYN® using a batch size of 10L, (2) filtering the bulk solution through a filter with a porosity size of at least 5 μm, and (3) remove the galactomannan content from the bulk solution by the ultrafiltration / diafiltration technique. The sampling process was conducted during the ultrafiltration treatment of the bulk solution for up to 10 concentrations (10X) and six diafiltration process (6DV).

Bulk Formulation A bulk solution of a Zosyn® bulk product development batch was formulated at a concentration of 250 g / mL. Piperacillin and 31.25 mG / mL of Tazobactam with an excess of 12% Piperacillin to direct the reaction to be complete. The starting material of piperacillin monohydrate (PMH), lot number 2000084742, which was tested as positive for galactomannan (GM) (using the EIA Bio-Rad Platelia ™ kit) was used in this study. Sodium bicarbonate (limit reagent) was added on a stoichiometric basis. The total batch size was 10L. The weight data are summarized in the table of starting materials. The bulk formulation was performed well, as expected. For protocol purposes, the bulk solution of the product was not brought to final volume (Qs). The reaction was considered complete since the solution reached a pH of 6.0 (acceptable pH limit of 6.8 or less). A volume of bulk product of about 8 liters (8 L) was obtained before for the qs. For the purpose of this study, the bulk solution of the product was not brought to the final volume. The table of starting materials summarizes the different ingredients of the formulation used for the manufacture of the experimental batch.

Filtration Once the reaction was completed and before reaching the final volume of 10L, the bulk product was filtered through a nylon membrane filter with a porosity size of 0.2 μm (CUNO® LifeASSURE ™ capsule).

Ultrafiltration / Diafiltration Process The ultrafiltration (UF) filtering process was conducted using a Omega® 5-kD membrane (part OS005G02). The previous membrane size was selected since the GM removal efficiency is greater than 1, 3 and 10kD membranes.

A total volume of the 6L bulk solution of ZOSYN® was used to evaluate the operational efficiency of the filtration system. The UF system operated with a feed pressure of 37 psi (42 psi, maximum pressure) and a retention pressure of 35 psi (39 psi, maximum pressure). During ultrafiltration, the pooled clustered samples were taken at 2X, 4X, 8X, and 10X concentrations. Once the 10X concentration was achieved, a recovery yield of 96% for Piperacillin and 86% for Tazobactam were obtained as shown in Table A.

A filtering process by diafiltration followed and was executed by completing six diafiltration volumes (2DV, 4DV, 5DV, and 6DV). The data collected showed that, after four volume of diafiltration (4DV), a recovery yield of 100% is obtained for both Piperacillin and Tazobactam as shown in Table B.

Concurrently, the GM detection test was carried out on each sample included in Tables A and B. The results obtained for the GM detection test are shown in Table C.

Note: 0.718, Average OD cut control The samples of the permeate grouped gave negative results for galactomannan. All the test results were well within the established specifications.

Claims (13)

1. A pharmaceutical composition comprising effective amounts of (a) piperacillin or a pharmaceutically acceptable salt thereof, (b) Tazobactam or a pharmaceutically acceptable salt thereof substantially free of galactomannan or a pharmaceutically acceptable salt thereof.

2. A pharmaceutical composition according to claim 1 wherein the piperacillin is sodium piperacillin.

3. A pharmaceutical composition according to claim 1 or claim 2 wherein the Tazobactam is tazobactam sodium.

4. A pharmaceutical composition according to any one of claims 1 to 3 wherein the composition is a lyophilized powder.

5. A method for the treatment or control of bacterial infections in a mammal wherein the method comprises administering to said mammal a therapeutically effective amount of the pharmaceutical composition of claim 1.

6. A process for preparing a lyophilized pharmaceutical composition substantially free of galactomannan comprising the steps of: a) dissolving piperacillin, and tazobactam in an aqueous solvent forming a solution and adjusting the pH to about 6.5; b) filter the solution through a filter cutter; c) Collect a filtrate; d) Cool the filtrate to a temperature below -35 ° C in a lyophilizer; e) Evacuate the lyophilizer at a pressure of about 300 μm Hg (micrometers of mercury) (40 pass) and heat the lyophilizer to about + 5 ° C; f) Maintaining the temperature and pressure for a sufficient time to remove the water from the aqueous solvent formed a lyophilized solid; g) Dry the lyophilized solid at about + 45 ° C.

7. A pharmaceutical composition according to claim 6 wherein the cut filter has a molecular weight of about 3 kD to about 10 kD.

8. A pharmaceutical composition according to claim 6 wherein the cut filter has a molecular weight of about 3 kD.

9. A pharmaceutical composition according to claim 6 wherein the cut filter has a molecular weight of about 5 kD.

10. A pharmaceutical composition according to claim 6 further comprising an index of experimental samples of the collected filtrate that is less than 0.5.

11. A process for the manufacture of a pharmaceutical composition in the form of a powder that can be reconstituted by the addition of a compatible reconstitution diluent before administration to a mammal or in the form of a frozen composition that when thawed can be diluted with a compatible diluent prior to administration to a mammal said process comprises freezing or freeze drying a solution substantially free of galactomannan containing effective amounts of (a) piperacillin or a pharmaceutically acceptable salt thereof, (b) Tazobactam or a pharmaceutically salt acceptable of it or a pharmaceutically acceptable salt thereof in an aqueous carrier

12. A pharmaceutical composition comprising an effective amount of piperacillin substantially free of galactomannan or a pharmaceutically acceptable salt thereof.

13. A pharmaceutical composition according to claim 12 wherein the piperacillin is sodium piperacillin.