CEFTOLOZANE PHARMACEUTICAL COMPOSITIONS

TECHNICAL FIELD

This disclosure relates to pharmaceutical compositions comprising ceftolozane.

BACKGROUND

Ceftolozane is a cephalosporin antibacterial agent, also referred to as CXA-101, FR264205, or by chemical names such as (6R,7R)-3-[(5-amino-4-{[(2-aminoethyl)carbamoyl]amino}-1-methyl-1H-pyrazol-2-ium-2-yl)methyl]-7-({(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-[(1-carboxy-1-methylethoxy)imino]acetyl}amino)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, and 7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-{3-amino-4-[3-(2-aminoethyl)ureido]-2-methyl-1-pyrazolio}methyl-3-cephem-4-carboxylate.

The prior art describes a variety of ceftolozane salts. For example, U.S. Pat. No. 7,129,232 discloses ceftolozane hydrogen sulfate salt among other salts “with a base or an acid addition salt such as a salt with an inorganic base, for example, an alkali metal salt [e.g., sodium salt, potassium salt, etc.], an alkaline earth metal salt [e.g., calcium salt, magnesium salt, etc.], an ammonium salt; a salt with an organic base, for example, an organic amine salt [e.g., trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N′-dibenzylethylenediamine salt, etc.]; an inorganic acid addition salt [e.g., hydrochloride, hydrobromide, sulfate, hydrogen sulfate, phosphate, etc.]; an organic carboxylic or sulfonic acid addition salt [e.g., formate, acetate, trifluoroacetate, maleate, tartrate, citrate, fumarate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.]; and a salt with a basic or acidic amino acid [e.g., arginine, aspartic acid, glutamic acid, etc.].” Ceftolozane sulfate is a pharmaceutically acceptable ceftolozane salt of formula (I) that can be formulated for intravenous administration or infusion.

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Certain pharmaceutical compositions containing ceftolozane are useful as antibiotics for the treatment of certain serious infections, including serious complicated intra-abdominal infections and complicated urinary tract infections. The ceftolozane pharmaceutical compositions can be administered as intravenous antibacterial agents to treat these infection. The antibacterial activity of ceftolozane is believed to result from its interaction with penicillin binding proteins (PBPs) to inhibit the biosynthesis of the bacterial cell wall which acts to stop bacterial replication. Antibacterial pharmaceutical compositions can include a therapeutically effective unit dose of a pharmaceutically acceptable salt of ceftolozane formulated for intravenous administration.

As disclosed herein, ceftolozane was unstable in certain lyophilized pharmaceutical compositions initially evaluated for intravenous administration. In particular, a decrease in ceftolozane purity and the formation of multiple additional related substances were detected in certain initial ceftolozane pharmaceutical compositions by peaks in high purity liquid chromatography (HPLC) after stability testing. This testing pointed to the need to develop novel ceftolozane formulations providing increased ceftolozane stability. U.S. Pat. No. 7,129,232 discloses that “auxiliary substances” such as “stabilizing agents . . . and other commonly used additives” may be included in pharmaceutical compositions comprising ceftolozane or many other cephalosporin compounds “if needed.” However, the disclosure does not disclose a reduction in ceftolozane purity in pharmaceutical compositions containing ceftolozane sulfate, or the formation of additional related substances observed during stability testing. Nor does this disclosure provide guidance on the formation of ceftolozane pharmaceutical compositions to increase ceftolozane purity during stability testing or control the relative amounts of ceftolozane related substances detected by HPLC peak formation during stability testing.

In view of the above, there is a need for pharmaceutical preparations containing ceftolozane compounds having improved ceftolozane stability.

SUMMARY

The pharmaceutical antibiotic compositions can include ceftolozane sulfate obtained by a process comprising the steps of lyophilizing an aqueous solution containing ceftolozane and a stabilizing amount of sodium chloride, where the stabilizing amount of sodium chloride is about 125 to 500 mg of sodium chloride (including, e.g., 480-500 mg) per 1,000 mg ceftolozane active in the aqueous solution prior to lyophilization. As disclosed in the Examples herein, high performance liquid chromatography (HPLC) analysis conducted during stability testing of lyophilized ceftolozane pharmaceutical compositions comprising between 50 and 481 mg sodium chloride and 1,000 mg of ceftolozane revealed an improved ceftolozane stability in compositions formed from aqueous solutions having at least 125 mg sodium chloride. Unexpectedly, lyophilized ceftolozane compositions having 125 mg or more sodium chloride relative to the fixed amount of ceftolozane active prior to lyophilization were about 35-90% more stable than comparable ceftolozane compositions having less than 125 mg sodium chloride, as measured by total ceftolozane purity during a 7-day stability study in Example 3. In addition, lyophilized ceftolozane compositions having 125 mg or more sodium chloride relative to the fixed amount of 1,000 mg ceftolozane active in an aqueous solution prior to lyophilization produced lyophilized compositions having lower quantities of additional substances identified by peaks 1 and 7 having characteristic retention times measured by HPLC relative to the retention time of ceftolozane. Unless otherwise indicated, HPLC measurements reported herein refer to HPLC using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C. Using this system, the retention time of peaks 1 and 7 are about −0.1 and about 1.3 relative to ceftolozane (an “RRT of 0.1 and 1.3, respectively).

In particular, sodium chloride stabilized ceftolozane compositions described in Example 3 were characterized by about 37-94% less of the material of peak 1 and about 38-306% less of the material of peak 7 (measured by corresponding HPLC peak areas) than comparable ceftolozane compositions having less than 125 mg sodium chloride (e.g., see 7-day stability study in Example 3).

The disclosed sodium chloride stabilized ceftolozane compositions can be characterized by decrease in ceftolozane total purity is not greater than 4.06% after storing the pharmaceutical composition for seven days at 60° C., as determined by HPLC using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C. These sodium chloride stabilized ceftolozane compositions were characterized by an increase in the amount of the impurity represented by Peak 1 not greater than 1.83% after storing the pharmaceutical composition for seven days at 60° C., as determined by HPLC using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C., where Peak 1 has a retention time relative to ceftolozane of 0.1.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B are chromatograms of CXA-101 ceftolozane drug substance obtained from the lyophilization process of Example 1. The chromatograms were obtained according to the analytical method described in Example 2.

FIG. 2 is a diagram of a lyophilization process for the ceftolozane obtained according to the process described in Example 1.

FIG. 3 is a table (Table 1) of peaks for the ceftolozane prepared by the lyophilization process in Example 1 obtained by HPLC according to the analytical method of Example 2.

FIG. 4 is a table (Table 2) showing the composition of various additional ceftolozane pharmaceutical compositions in which the sodium chloride content is varied.

FIG. 5A is a table (Table 3) showing the total purity of ceftolozane in the pharmaceutical compositions of FIG. 4, as measured by HPLC peak area according to the analytical method of described in Example 2.

FIG. 5B is a graph showing the total purity of certain pharmaceutical compositions disclosed in FIG. 4, as measured by HPLC peak area.

FIG. 6A is a table (Table 4) showing the amount of material from characteristic peak 1 in the pharmaceutical compositions of FIG. 4, as measured by HPLC peak area according to the analytical method of described in Example 2.

FIG. 6B is a graph showing the amount of material from characteristic peak 1 in the pharmaceutical compositions of FIG. 4, as measured by HPLC peak area according to the analytical method of described in Example 2.

FIG. 7A is a table (Table 5) showing the amount of material from characteristic peak 3 in the pharmaceutical compositions of FIG. 4, as measured by HPLC peak area according to the analytical method of described in Example 2.

FIG. 7B is a graph showing the amount of material from characteristic peak 3 in the pharmaceutical compositions of FIG. 4, as measured by HPLC peak area according to the analytical method of described in Example 2.

FIG. 8A is a table (Table 6) showing the amount of material from characteristic peak 7 in the pharmaceutical compositions of FIG. 4, as measured by HPLC peak area according to the analytical method of described in Example 2.

FIG. 8B is a graph showing the amount of material from characteristic peak 7 in the pharmaceutical compositions of FIG. 4, as measured by HPLC peak area according to the analytical method of described in Example 2.

FIG. 9A is a table (Table 7) showing the formulation composition of the Co-Lyo-Combo Drug Product used in Example 4A.

FIG. 9B is a table (Table 8) showing impurity concentrations at time zero, one month and three months at 25° C./60% relative humidity of the Co-Lyo-Combo Drug Product.

FIG. 9C is a table (Table 9) showing impurity concentrations at time zero, one month and three months at 40° C./75% relative humidity of the Co-Lyo-Combo Drug Product.

FIG. 10A is a table (Table 10) showing the formulation composition of the Blend Drug Product used in Example 4B.

FIG. 10B is a table (Table 11) showing impurity concentrations at time zero, one month and three months at 25° C./60% relative humidity of the Blend Drug Product used in Example 4B.

FIG. 10C is a table (Table 12) showing impurity concentrations at time zero, one month and three months at 40° C./75% relative humidity of the Blend Drug Product used in Example 4B.

FIG. 11 is a table (Table 13) showing the composition of various ceftolozane pharmaceutical compositions in which the sodium chloride concentration is varied.

FIG. 12 is a table (Table 14) showing the purity of Ceftolozane in CXA-201 Compositions with varying amounts of sodium from sodium chloride at time zero, 1 day, 3 days and 7 days at at 30° C. and 60° C.

FIG. 13 is a table (Table 15) showing the HPLC area of Impurity of Peak 1 in CXA-201 Compositions with varying amounts of sodium from sodium chloride at time zero, 1 day, 3 days and 7 days at 30° C. and at 60° C.

FIG. 14 is a table (Table 16) showing the HPLC area of the Impurity at RRT 0.43 and Impurity Peak 3 in CXA-201 Compositions with varying amounts of sodium from sodium chloride at time zero, 1 day, 3 days and 7 days at 30° C. and at 60° C.

FIG. 15 is a table (Table 17) showing the HPLC area of Impurity of Peak 7 in CXA-201 Compositions with varying amounts of sodium from sodium chloride at time zero, 1 day, 3 days and 7 days at 30° C. and at 60° C.

FIG. 16 is a table (Table 18) showing the finished drug product unit composition of ceftolozane/tazolactam.

FIG. 17 is a table (Table 19) showing the primary container closure system for the ceftolozane/tazobactam unit product.

DETAILED DESCRIPTION

Ceftolozane sulfate can be stabilized in lyophilized pharmaceutical compositions by incorporation of a stabilizing-effective amount of an inorganic salt stabilizing agent in a solution that can be lyophilized to obtain a lyophilized composition containing stabilized ceftolozane. The stabilizing-effective amount of a sodium chloride inorganic salt stabilizing agent is preferably 125 mg to 500 mg (preferably about 480-500 mg) of sodium chloride per 1,000 mg of ceftolozane active.

In particular, pharmaceutical compositions comprising ceftolozane and stabilizing amount of sodium chloride can be obtained by lyophilization of a solution comprising a stabilizing-effective amount of sodium chloride and ceftolozane sulfate. Alternatively, they can be obtained by other methods. As is known to those skilled in the art, lyophilization is a process of freeze-drying in which water is sublimed from a frozen solution of one or more solutes. Specific methods of lyophilization are described in Remington's Pharmaceutical Sciences, Chapter 84, page 1565, Eighteenth Edition, A. R. Gennaro, (Mack Publishing Co., Easton, Pa., 1990). A pharmaceutical composition comprising ceftolozane can be prepared by adding a stabilizing amount of sodium chloride in a fixed ratio to ceftolozane in an aqueous solution prior to lyophilization, then lyophilizing the solution to obtain a lyophilized composition comprising sodium chloride and ceftolozane.

In particular, the pharmaceutical antibiotic compositions can include ceftolozane sulfate obtained by a process comprising the steps of lyophilizing an aqueous solution containing ceftolozane and a stabilizing amount of sodium chloride, where the stabilizing amount of sodium chloride is about 125 to 500 mg (preferably, 480-500 mg) of sodium chloride per 500 mg ceftolozane active in the aqueous solution prior to lyophilization. The pharmaceutical compositions comprising ceftolozane disclosed herein can be prepared by adding 125 to 500 mg (preferably, 480-500 mg) sodium chloride per 1000 mg of ceftolozane prior to lyophilization (or spray drying). For example, the pharmaceutical compositions can be obtained by a method comprising the steps of adding a stabilizing amount (e.g., 125 to 500 mg [more preferably, 480-500 mg] sodium chloride per 1000 mg of ceftolozane active) followed by lyophilizing (or spray drying) the composition comprising the sodium chloride and ceftolozane. In one aspect (e.g., Example 1), ceftolozane and a stabilizing amount of sodium chloride can be dissolved in an aqueous solution that can be lyophilized (or spray drying) to obtain a ceftolozane pharmaceutical composition.

The pharmaceutical compositions may comprise other additional components including stabilizers, pH adjusting additives (e.g., buffers) and the like. Non-limiting examples of these additives include sodium chloride, citric acid and L-arginine. For example, the pharmaceutical antibiotic compositions can include ceftolozane sulfate obtained by a process comprising the steps of lyophilizing an aqueous solution containing ceftolozane sulfate with a stabilizing amount of sodium chloride (e.g., 125 to 500 mg [more preferably, 480-500 mg] of sodium chloride per 1,000 mg ceftolozane active), with L-arginine and/or citric acid in the aqueous solution prior to lyophilization. The use of sodium chloride results in greater ceftolozane stability, while an amount of L-arginine can be used that is effective to adjust pH and to increase the solubility of ceftolozane, and citric acid can be included in an amount effective to reduce or prevent discoloration of the product, due to its ability to chelate metal ions. The aqueous solution can be subsequently lyophilized (or spray dried) to obtain a stabilized lyophilized ceftolozane sulfate composition comprising ceftolozane sulfate, sodium chloride, L-arginine and citric acid.

The pharmaceutical composition can also be a Ceftolozane/Tazobactam for Injection Drug Product, 1000 mg/500 mg. It is presented as a combination of two sterile active powders in a single vial intended for reconstitution and intravenous infusion.

The drug product is first prepared by converting ceftolozane sulfate drug substance to a sterile drug product intermediate (DPI) powder with excipients citric acid, sodium chloride and L-arginine. This is commonly done by lyophilization, as described above. Tazobactam sodium drug substance is presented as a sterile powder without any excipients. The tazobactam sodium drug substance is typically lyophilized (or spray dried). The drug product is then prepared by aseptically filling the two powders (e.g., the two separately lyophilized drug powders or spray dried powders) sequentially into a single vial.

Each vial of ceftolozane/tazobactam for injection contains approximately 2255 mg ceftolozane sterile DPI powder that contains 1147 mg ceftolozane sulfate, which is equivalent to 1000 mg ceftolozane free base, as well as approximately 537 mg tazobactam sodium sterile drug substance, equivalent to 500 mg tazobactam free acid. At the time of administration, the vial is reconstituted with 10 mL vehicle, 5% Dextrose Injection USP, sterile Water for Injection or 0.9% Sodium Chloride Injection USP, then the vial contents further diluted in an infusion bag of 0.9% Sodium Chloride Injection USP or 5% Dextrose Injection USP, for administration. The constituents are shown in Table 18, FIG. 16.

The primary container-closure system is a Type 1 20 mL molded glass vial with 20 mm neck finish. The vial is sealed by a 20 mm rubber stopper and 20 mm plastic flip-cap seal with aluminum ferrule. The primary contain-closure system used for the ceftolozane/tazobactam unit product is summarized in Table 19, FIG. 17.

In other embodiments, pharmaceutical compositions comprising ceftolozane can be obtained by methods that include the steps of: (1) adding a stabilizing amount of sodium chloride to ceftolozane (or preparing an aqueous solution of sodium chloride and ceftolozane) optionally followed by co-lyophilizing or spray drying the ceftolozane and sodium chloride (or aqueous solution thereof); and (2) combining the product of step (1) with other components. For example, the product of step (1) can be combined with a β-lactamase inhibitor, such as tazobactam (CAS#: 89786-04-9), avibactam (CAS#1192500-31-4), Sulbactam (CAS#68373-14-8) and/or clavulanate (CAS#58001-44-8). The beta lactamase inhibitor can be included in a crystalline or amorpous form, such as a lyophilized tazobactam or crystalline tazobactam (e.g., U.S. Pat. Nos. 8,476,425 and 5,763,603) to obtain the pharmaceutical composition.

The stabilizing effect of sodium chloride on ceftolozane can be measured by high performance liquid chromatography (HPLC) analysis, for example by detecting the ratio of peak areas obtained for ceftolozane compared to peaks for other substances. Various ceftolozane compositions having 50-481 mg of sodium chloride per 1,000 mg ceftolozane active as described in Table 2 (FIG. 4) were tested for stability as described in Example 3. The stability testing of lyophilized ceftolozane pharmaceutical compositions (Example 3) revealed improved ceftolozane stability in compositions formed from aqueous solutions having at least 125 mg sodium chloride. Unless otherwise indicated, HPLC measurements reported herein are obtained using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C.

Unexpectedly, lyophilized ceftolozane compositions having 125 mg more sodium chloride relative to the fixed amount of ceftolozane prior to lyophilization were about 35-90% more stable than comparable ceftolozane compositions having less than 125 mg sodium chloride, as measured by total ceftolozane purity during a 7-day stability study in Example 3. Referring to the data in Table 3 (FIGS. 5A and 5B), the total purity of ceftolozane (measured by HPLC according to Example 2) was measured by the % decrease in the ceftolozane HPLC peak during the 7-day stability test. In particular, samples obtained by lyophilizing compositions containing 125 mg, 190 mg and 481 mg of sodium chloride per 1,000 mg of ceftolozane active showed a decrease in ceftolozane that was at least about 35% less than reductions in ceftolozane observed for formulations containing 50 mg or 75 mg sodium chloride per 1,000 mg ceftolozane active (e.g., the % decrease in ceftolozane for the sample containing 75 mg sodium chloride was about 35% greater than the comparable % decrease in ceftolozane for the sample containing 190 mg sodium chloride). In addition, samples obtained by lyophilizing compositions containing 125 mg, 190 mg and 481 mg of sodium chloride per 1,000 mg of ceftolozane active showed a decrease in ceftolozane that was up to about 90% less than reductions in ceftolozane observed for forulations containing 50 mg or 75 mg sodium chloride per (e.g., the % decrease in ceftolozane for the sample containing 50 mg sodium chloride was about 90% greater than the comparable % decrease in ceftolozane for the sample containing 481 mg sodium chloride).

In addition, lyophilized ceftolozane compositions having 125 mg more sodium chloride relative to the fixed amount of 1,000 mg ceftolozane active in an aqueous solution prior to lyophilization produced lyophilized compositions having lower quantities of additional substances identified by peaks 1 and 7 having characteristic retention times measured by HPLC (see Table 1, FIG. 3, indicating retention times of about 0.1 for peak 1 and about 1.3 for peak 7 relative to celftolozane measured according to the HPLC method of Example 2). In particular, these sodium chloride stabilized ceftolozane compositions were characterized by about 37-94% less of the material of peak 1 and about 38-306% less of the material of peak 7 (measured by corresponding HPLC peak areas) than comparable ceftolozane compositions having less than 125 mg sodium chloride (e.g., see 7-day stability study in Example 3).

Referring to the data in Table 4 (FIGS. 6A and 6B), the amount of the composition of peak 1 (measured by HPLC according to Example 2) was measured by the % increase in the peak 1 HPLC peak during the 7-day stability test of Example 3. In particular, samples obtained by lyophilizing compositions containing 125 mg, 190 mg and 481 mg of sodium chloride per 1,000 mg of ceftolozane active showed at least a 37% reduction in the amount of the peak 1 composition observed for these formulations containing at least 125 mg sodium chloride per 1,000 mg ceftolozane active, compared to the compositions obtained by lyophilizing 50 mg or 75 mg sodium chloride per 1,000 mg of ceftolozane active (e.g., the % increase in peak 1 for the sample containing 75 mg sodium chloride was about 37% greater than the comparable % decrease in ceftolozane for the sample containing 190 mg sodium chloride). In addition, samples obtained by lyophilizing compositions containing 125 mg, 190 mg and 481 mg of sodium chloride per 1,000 mg of ceftolozane active showed up to a 94% reduction in the amount of the peak 1 composition observed for these formulations containing at least 125 mg sodium chloride per 1,000 mg ceftolozane active, compared to the compositions obtained by lyophilizing 50 mg or 75 mg sodium chloride per 1,000 mg of ceftolozane active (e.g., the % increase in peak 1 for the sample containing 50 mg sodium chloride was about 94% greater than the comparable % decrease in ceftolozane for the sample containing 481 mg sodium chloride).

Referring to the data in Table 5 (FIGS. 7A and 7B), the amount of the composition of peak 3 (measured by HPLC according to Example 2) was measured by the % increase in the peak 1 HPLC peak during the 7-day stability test of Example 3. In particular, samples obtained by lyophilizing compositions containing 125 mg and 190 mg of sodium chloride per 1,000 mg of ceftolozane active showed at least a 269% reduction in the amount of the peak 3 composition observed for these formulations containing at least 125 mg sodium chloride per 1,000 mg ceftolozane active, compared to the compositions obtained by lyophilizing 50 mg or 75 mg sodium chloride per 1,000 mg of ceftolozane active (e.g., the % increase in peak 3 for the sample containing 50 mg sodium chloride was about 269% greater than the comparable % decrease in ceftolozane for the sample containing 190 mg sodium chloride). In addition, samples obtained by lyophilizing compositions containing 125 mg or 190 mg of sodium chloride per 1,000 mg of ceftolozane active showed up to a 333% reduction in the amount of the peak 3 composition observed for these formulations containing at least 125 mg sodium chloride per 1,000 mg ceftolozane active, compared to the compositions obtained by lyophilizing 50 mg or 75 mg sodium chloride per 1,000 mg of ceftolozane active (e.g., the % increase in peak 3 for the sample containing 75 mg sodium chloride was about 333% greater than the comparable % decrease in ceftolozane for the sample containing 125 mg sodium chloride). The ceftolozane sample containing 481 mg of sodium chloride had a higher amount of the composition of peak 3 than the ceftolozane samples containing 50 mg or 75 mg of sodium chloride.

Referring to the data in Table 6 (FIGS. 8A and 8B), the amount of the composition of peak 7 (measured by HPLC according to Example 2) was measured by the % increase in the peak 7 HPLC peak during the 7-day stability test of Example 3. In particular, samples obtained by lyophilizing compositions containing 125 mg, 190 mg and 481 mg of sodium chloride per 1,000 mg of ceftolozane active showed at least a 38% reduction in the amount of the peak 7 composition observed for these formulations containing at least 125 mg sodium chloride per 1,000 mg ceftolozane active, compared to the compositions obtained by lyophilizing 50 mg or 75 mg sodium chloride per 1,000 mg of ceftolozane active (e.g., the % increase in peak 7 for the sample containing 75 mg sodium chloride was about 38% greater than the comparable % decrease in ceftolozane for the sample containing 125 mg sodium chloride). In addition, samples obtained by lyophilizing compositions containing 125 mg, 190 mg and 481 mg of sodium chloride per 1,000 mg of ceftolozane active showed up to a 306% reduction in the amount of the peak 7 composition observed for these formulations containing at least 125 mg sodium chloride per 1,000 mg ceftolozane active, compared to the compositions obtained by lyophilizing 50 mg or 75 mg sodium chloride per 1,000 mg of ceftolozane active (e.g., the % increase in peak 7 for the sample containing 50 mg sodium chloride was about 306% greater than the comparable % decrease in ceftolozane for the sample containing 481 mg sodium chloride).

Accordingly, preferred pharmaceutical compositions contain ceftolozane sulfate having an improved stability as a decrease in the rate of ceftolozane purity and/or a decrease in the rate of formation of substances characterized by HPLC peaks 1 and 7 identified during a 7-day stability study in Example 3. The preferred ceftolozane pharmaceutical compositions comprise a stabilizing amount of sodium chloride (e.g., 125 to 500 mg of sodium chloride per 1000 mg of ceftolozane). Certain preferred compositions demonstrate improved ceftolozane purity (e.g., Table 3 in FIG. 5A) and chemical stability (e.g., with respect to the composition of HPLC peak 1 in Table 4, FIG. 6A) compared pharmaceutical compositions comprising ceftolozane with comparatively less sodium chloride.

Pharmaceutical compositions comprising ceftolozane can be formulated to treat infections by parenteral administration (including subcutaneous, intramuscular, and intravenous) administration. Pharmaceutical compositions may additionally comprise excipients, stabilizers, pH adjusting additives (e.g., buffers) and the like. Non-limiting examples of these additives include sodium chloride, citric acid and L-arginine. For example, the use of sodium chloride results in greater stability; L-arginine is used to adjust pH and to increase the solubility of ceftolozane; and citric acid is used to prevent discoloration of the product, due to its ability to chelate metal ions. In one particular embodiment, the pharmaceutical compositions described herein are formulated for administration by intravenous injection or infusion. Pharmaceutical antibiotic compositions can include ceftolozane sulfate and stabilizing amount of sodium chloride (e.g., 125 to 500 mg of sodium chloride per 1,000 mg ceftolozane active) in a lyophilized unit dosage form (e.g., powder in a vial). The unit dosage form can be dissolved with a pharmaceutically acceptable carrier, and then intravenously administered. In another aspect, pharmaceutical antibiotic compositions can include ceftolozane sulfate obtained by a process comprising the steps of lyophilizing an aqueous solution containing ceftolozane and a stabilizing amount of sodium chloride, where the stabilizing amount of sodium chloride is about 125 to 500 mg of sodium chloride per 1,000 mg ceftolozane active in the aqueous solution prior to lyophilization.

In one aspect, provided herein is a method for the treatment of bacterial infections in a mammal, comprising administering to said mammal a therapeutically effective amount of a pharmaceutical composition prepared according to the methods described herein. A method for the treatment of bacterial infections in a mammal can comprise administering to said mammal a therapeutically effective amount of a pharmaceutical composition comprising ceftolozane sulfate and sodium chloride. Non-limiting examples of bacterial infections that can be treated by the methods of the invention include infections caused by: aerobic and facultative gram-positive microorganisms (e.g., Staphylococcus aureus, Enterococcus faecalis, Staphylococcus epidermidis, Streptococcus agalactiae, Streptococcus pneumonia, Streptococcus pyogenes, Viridans group streptococci), aerobic and facultative gram-negative microorganisms (e.g., Acinetobacter baumanii, Escherichia coli, Haemophilus influenza, Klebsiella pneumonia, Pseudomonas aeruginosa, Citrobacter koseri, Moraxella catarrhalis, Morganella morganii, Neisseria gonorrhoeae, Proteus mirabilis, Proteus vulgaris, Serratia marcescens, Providencia stuartii, Providencia rettgeri, Salmonella enterica), gram-positive anaerobes (Clostridium perfringens), and gram-negative anaerobes (e.g., Bacteroides fragilis group (e.g., B. fragilis, B. ovatus, B. thetaiotaomicron, and B. vulgates), Bacteroides distasonis, Prevotella melaminogenica). In certain embodiments of the methods described herein, bacterial infection is associated with one or more of the following conditions: complicated intra-abdominal infections, complicated urinary tract infections (cUTIs) and pneumonia (e.g., community-acquired, or nosocomial pneumonia). Community-acquired pneumonia (moderate severity only) can include infections caused by piperacillin-resistant, beta-lactamase producing strains of Haemophilus influenza. Nosocomial pneumonia (moderate to severe) caused by piperacillin-resistant, beta-lactamase producing strains of Staphylococcus aureus and by Acinetobacter baumanii, Haemophilus influenzae, Klebsiella pneumoniae, and Pseudomonas aeruginosa.

As used herein, “treating”, “treat” or “treatment” describes the management and care of a patient for the purpose of combating a disease, condition, or disorder and includes the administration of a pharmaceutical composition of the present invention to alleviate the symptoms or complications of a disease, condition or disorder, or to reduce the extent of the disease, condition or disorder. The term “treat” can also include treatment of a cell in vitro or an animal model.

By a “therapeutically effective amount” of a compound of the invention is meant a sufficient amount of the compound to treat the disorder (e.g., bacterial infection). The specific therapeutically effective amount that is required for the treatment of any particular patient or organism (e.g., a mammal) will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound or composition employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts (see, for example, Goodman and Gilman's, “The Pharmacological Basis of Therapeutics”, Tenth Edition, A. Gilman, J. Hardman and L. Limbird, eds., McGraw-Hill Press, 155-173, 2001, which is incorporated herein by reference in its entirety). The therapeutically effective amount for a given situation can be readily determined by routine experimentation and is within the skill and judgment of the ordinary clinician.

As used herein, “125-500 mg sodium chloride per 1000 mg of ceftolozane” refers to a ratio of sodium chloride to ceftolozane active. The phrase “125-500 mg sodium chloride per 1000 mg of ceftolozane” includes “62.5 to 250 mg sodium chloride per 500 mg of ceftolozane active” and other similar weight ratios. In addition, “1,000 mg of ceftolozane as ceftolozane sulfate” refers to an amount of ceftolozane sulfate effective to provide 1,000 mg of ceftolozane. The amount of sodium per gram of ceftolozane activity in a pharmaceutical composition containing ceftolozane sulfate and sodium chloride can be calculated using the relevant molecular weights of ceftolozane, ceftolozane sulfate, sodium chloride and sodium.

As used herein, “1000 mg ceftolozane” refers to an amount of ceftolozane that is considered a bioequivalent by the United States Food and Drug Administration (FDA), i.e. for which 90% CI of the relative mean Cmax, AUC(0-t) and AUC(0-∞) is within 80.00% to 125.00% of the reference formulation in the fasting state (see: “Guidance for Industry: Bioavailability and Bioequivalence Studies for Orally Administered Drug Products—General Considerations”. Center for Drug Evaluation and Research, United States Food and Drug Administration, 2003).

“Ceftolozane active” refers to the active portion of a salt form of ceftolozane, i.e., the free base form of ceftolozane.

In another aspect, the disclosed sodium chloride stabilized ceftolozane compositions can be characterized by decrease in ceftolozane total purity is not greater than 3.7% after storing the pharmaceutical composition for seven days at 60° C., as determined by HPLC using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C. In another aspect, the disclosed sodium chloride stabilized ceftolozane compositions can be characterized by decrease in ceftolozane total purity is not greater than 4.2% after storing the pharmaceutical composition for seven days at 60° C., as determined by HPLC using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C. In another aspect, the disclosed sodium chloride stabilized ceftolozane compositions can be characterized by decrease in ceftolozane total purity is not greater than 4.5% after storing the pharmaceutical composition for seven days at 60° C., as determined by HPLC using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C. In another aspect, the disclosed sodium chloride stabilized ceftolozane compositions can be characterized by decrease in ceftolozane total purity is not greater than 5.0% after storing the pharmaceutical composition for seven days at 60° C., as determined by HPLC using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C.

In another aspect, the disclosed sodium chloride stabilized ceftolozane compositions were characterized by an increase in the amount of the impurity represented by Peak 1 not greater than 1.8% after storing the pharmaceutical composition for seven days at 60° C., as determined by HPLC using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C., where Peak 1 has a retention time relative to ceftolozane of 0.1. In another aspect, the disclosed sodium chloride stabilized ceftolozane compositions were characterized by an increase in the amount of the impurity represented by Peak 1 not greater than 2.0% after storing the pharmaceutical composition for seven days at 60° C., as determined by HPLC using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C., where Peak 1 has a retention time relative to ceftolozane of 0.1. In another aspect, the disclosed sodium chloride stabilized ceftolozane compositions were characterized by an increase in the amount of the impurity represented by Peak 1 not greater than 2.2% after storing the pharmaceutical composition for seven days at 60° C., as determined by HPLC using a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of 45° C., where Peak 1 has a retention time relative to ceftolozane of 0.1.

Illustrative Examples of Selected Embodiments of the Invention
Example 1
Manufacturing Procedure of Bulk (Tray) Lyophilized Ceftolozane

There are four main steps in the manufacture of CXA-101 bulk drug product: dissolution, sterile filtration, bulk lyophilization, and packaging into Sterbags®. These four main steps are composed of a total of 20 minor steps. The CXA-101 bulk drug product manufacturing process is presented below.

I. Dissolution

1. The prescribed amount of water for injection (“WFI”) is charged into the dissolution reactor.

2. A prescribed amount of citric acid is added.

3. The solution is cooled at 5° C. to 10° C.

4. A prescribed amount of CXA-101 drug substance is added to the solution.

5. A prescribed amount of L-arginine is slowly added to the solution.

6. A check for complete dissolution is performed. Solution pH is verified to be in the target range of 6.5 to 7.0.

7. A prescribed amount of sodium chloride is added to the solution, wherein the preferred amount of sodium chloride is 125-500 mg of sodium chloride per 1000 mg of ceftolozane active.

8. A check for complete dissolution is performed. Solution pH is verified to be in the target range of 6.0 to 7.0. If the pH is out of this range adjust with either L-Arginine or citric acid.

9. WFI is added to bring the net weight to 124.4 kg and the solution is mixed well.

10. Samples are withdrawn for testing of final pH.

II. Sterile Filtration

11. The solution is passed through the filter (pore size 0.45 μm) followed by double filters (pore size 0.22 μm) onto a shelf on the Criofarma lyophilizer.

12. The line is washed with WFI.

13. The washing solution is passed from Step 12 through sterile filtration.

III. Bulk Lyophilization

14. The washing solution is loaded onto a separate shelf in the lyophilizer (and later discarded).

15. The solution is lyophilized until dry.

16. The product shelf is cooled to 20° C.±5° C.

IV. Packaging into Sterbags®

17. The lyophilized bulk drug product powder is milled.

18. The milled powder is sieved.

19. The sieved powder is blended for 30 minutes.

20. The powder is then discharged into Sterbags®

Prefiltration and Sterile-Filtration

Filtrate the compounded solution with a sterile tilter-set which consists of a 0.2 um polyvinylidene fluoride membrane filter (Durapore®, Millipore) and a 0.1 um polyvinylidene fluoride membrane filter (Durapore®, Millipore) connected in tandem. Confirm the integrity of each filter before and after the filtration. Take approximately 100 mL of the filtrate in order to check bioburden.

Filter the prefiltered compounded solution through a sterile filter-set which consists of a 0.2 um polyvinylidene fluoride membrane filter and a 0.1 um polyvinylidene fluoride membrane filter connected in tandem, and introduce the final filtrate into an aseptic room. Confirm the integrity of each filter before and after the filtration.

Processing of Vial, Stopper and Flip-Off Cap

Wash a sufficient quantity of 28 mL vials with water for injection and sterilize the washed vials by a dry-heat sterilizer. Then transfer the sterilized vials into a Grade A area located in an aseptic room.

Wash a sufficient quantity of stoppers with, water for injection. Sterilize and dry the washed stoppers by steam sterilizer. Then transfer the sterilized stoppers into a Grade A area located in an aseptic room.

Sterilize a sufficient quantity of flip-off caps by steam sterilizer. Then transfer the sterilized flip-off caps into a Grade A or B area located in an aseptic room.

Filling and Partially Stoppering

Adjust the fill weight of the filtered compounded solution to 11.37 g (corresponds to 10 mL of the compounded solution), then start filling operation. Check the filled weight in sufficient frequency and confirm it is in target range (11.37 g±1%, 11.26 to 11.43 g). When deviation from the control range (11.37 g±2%, 11.14 to 11.59 g) is occurred, re-adjust the filling weight.

Immediately after a vial is filled, partially stopper the vial with a sterilized stopper. Load the filled and partially stoppered vials onto the shelves of a lyophilizer aseptically.

Lyophilization to Crimping, Visual Inspection, Labeling and Packaging

After all filled and partially stoppered vials are loaded into a lyophilizer, start the lyophilization program shown in FIG. 2. Freeze the loaded vials at −40° C. and keep until all vials freeze. Forward the program to primary drying step (shelf temperature; −20° C., chamber pressure; 100 to 150 mTorr). Primary drying time should be determined by monitoring the product temperature. Forward the program to secondary drying step (shelf temperature; 30° C., chamber pressure; not more than 10 mTorr) after completion of the primary drying step. After all vials are dried completely, return the chamber pressure to atmospheric pressure with sterilized nitrogen. Then stopper vials completely.

Unload the lyophilized vials from the chamber and crimp with sterilized flip-off caps.

Subject all crimped vials to visual inspection and label and package all passed vials.

Example 2
Analytical HPLC Method
A. Operative Conditions

Column
Develosil ODS-UG-5; 5 μm, 250 × 4.6 mm

(Nomura Chemical, Japan)

Mobile phase
Sodium Perchlorate Buffer Solution

(PH 2.5)/CH₃CN 90:10 (vlv)

Flow rate
1.0 mL/min

Wavelength
254 nm

Injection volume
10 μL

Oven Temperature
45° C.

Run Time
85 minutes

Gradient Profile:

Time (min)
A %
B %

0
75
25

30
70
30

60
0
100

85
0
100

85.1
75
25

110
75
25

B. Mobile Phase Preparation.

Sodium Perchlorate Buffer Solution was made by dissolving 14.05 g of sodium perchlorate Monohydrate in 1000.0 mL of water followed by adjusting pH to 2.5 with diluted perchloric acid (1 in 20).

Mobile Phase was then made by mixing Sodium Perchlorate Buffer Solution (pH 2.5) and acetonitrile in the ratio 90:10 (v/v).

Sodium Acetate Buffer Solution pH 5.5 (Diluent) was made by dissolving 1.36 g of sodium acetate trihydrate in 1000.0 mL of water followed by adjusting to pH 5.5 with diluted acetic acid (1 in 10).

C. Sample Preparation.

Sample solution: dissolve 20.0 mg, exactly weighed, of Sample, in 20.0 mL of water (Prepare just before injection into HPLC system).

System Suitability Solution (1%): take 1.0 mL of the Sample Solution (use first sample if more are present) and transfer into a 100.0 mL volumetric flask, dilute with water to volume and mix.

D. HPLC Analysis Procedure

1. Inject Blank (water)

2. Inject System Suitability Solution and check for tailing factor and theoretical plate number for CXA-101 peak:

- The tailing factor must not be greater than 1.5
- Theoretical plates number must not be less than 10000

3. Inject Sample Solution

4. Inject System Suitability Solution and check for tailing factor and theoretical plate number for CXA-101 peak.

- The tailing factor must not be greater than 1.5
- Theoretical plates number must not be less than 10000
  
  5. Identify the peaks of Related Substances in the Sample chromatogram based on the reference chromatogram reported in FIGS. 1A and 1B or, alternatively, on the basis of the RRT values reported in Table 1 (FIG. 3), with a retention time of about 0.14 relative to ceftolozane is believed to have the chemical structure of formula (II):

embedded image

E. Calculations

I. Report for each related substance its amount as expressed by area percent.

$C_{i} = \frac{A_{i} \times 100}{A_{t} + \sum A_{i}}$

wherein:

C_i=Amount of related substance i in the Sample, area %

A_i=Peak area of related substance i in the Sample chromatogram

A_t=Area of CXA-101 peak in the Sample chromatogram

A_t+Σ A_i=Total peaks area in the Sample chromatogram

Consider as any Unspecified Impurity, each peak in the chromatogram except CXA-101, peaks from 1 to 11 and every peak present in the blank chromatogram and report the largest.

II. Report the total impurities content as expressed by the following formula:

$C_{T} = \frac{A_{i} \times 100}{A_{t} + \sum A_{i}}$

wherein:

C_T=total impurities content in the Sample, area %

A_t=area of CXA-101 peak in the sample chromatogram

Σ A_i=total peak areas of impurities in the sample chromatogram

Example 3
The Amount of Sodium Chloride can be Selected to Stabilize Ceftolozane in Pharmaceutical Compositions

Multiple stability studies were performed on ceftolozane sulfate, wherein the effect of varying amounts of sodium chloride on the stability of ceftolozane was examined.

The amount of sodium per mg of sodium chloride can be calculated (as known to one of ordinary skill in the art) based on the relative molar weight ratio of sodium and sodium chloride (e.g., 50 mg sodium chloride contains about 20 mg of sodium, etc).

The amount of ceftolozane in ceftolozane sulfate can similarly be calculated based on the respective molecular molar weights of ceftolozane and ceftolozane sulfate (e.g., 1,147 mg ceftolozane sulfate contains about 1,000 mg of ceftolozane). Accordingly, a composition comprising about 1,147 mg ceftolozane sulfate and 480 mg of sodium chloride also contains 480 mg of sodium chloride per 1,000 mg of ceftolozane.

A. CXA-101 Purity Increases in Compositions Having at Least about 125 Mg NaCl/1,000 Mg Ceftolozane Active

A stability study was carried out at 60° C. as described herein. Sodium chloride content in test samples is described in Table 2 (FIG. 4). The samples were formulated with 481, 190, 125, 75, and 50 mg sodium chloride per 1 g of ceftolozane active.

FIG. 5A is Table 3 with data for total purity of ceftolozane measured by HPLC during the 7-day stability test using the HPLC method in Example 2, with the data plotted in the graph of FIG. 5B.

B. The Amount of Substances Identified by HPLC Peaks 1 and 7 Decreases in Compositions Having at Least about 125 Mg NaCl/1,000 Mg Ceftolozane Active

A stability study was carried out at and 60° C. as described herein. Sodium chloride content in test samples is described in Table 2 (FIG. 4). The samples were formulated with 481, 190, 125, 75, and 50 mg sodium chloride per 1 g of ceftolozane active.

Stability data for amounts of additional substances in the ceftolozane compositions from Table 2 (FIG. 4) as measured by peaks 1, 3 and 7 by HPLC (according to Example 2) are summarized in Tables 4-6, FIGS. 6A, 7A and 8A. The data are also plotted in FIGS. 6B, 7B, and 8B to show trends of total purity, peak 1, RRT 0.43+peak 3, and peak 7 with respect to sodium chloride (NaCl), respectively. FIG. 5A is Table 3 with total purity of ceftolozane measured by HPLC during the 7-day stability test, with the data plotted in the graph of FIG. 5B.

Example 4
The Amount of Sodium Chloride can be Selected to Stabilize Ceftolozane in Pharmaceutical Compositions Comprising Ceftolozane and Tazobactam

A. Stabilized Ceftolozane Compositions Co-Lyophilized with Tazobactam

A composition comprising ceftolozane, sodium chloride and tazobactam was prepared by co-lyophilizing a stabilizing amount of sodium chloride, ceftolozane sulfate and tazobactam acid in amounts described in Table 7 (FIG. 9A) together in an aqueous solution to obtain a stabilized ceftolozane lyophilized composition (“Co-Lyophilized Combo Drug Product”). The components of the composition that was lyophilized to obtain the Co-Lyophilized Combo Drug Product is shown in Table 7 (FIG. 9A). Notably, this composition included about 484 mg of sodium chloride per 1,000 mg ceftolozane active provided as ceftolozane sulfate, and a weight ratio of about 2:1 between the CXA101 and tazobactam acid.

The results of a stability study of the Co-Lyophilized Combo Drug Product are shown in Table 8 (FIG. 9B) and Table 9 (FIG. 9C) as representative examples that summarize the results at 25° C./RH=60% and 40° C./RH=75% after one month (T1) and three months (T2). Samples were analyzed using a HPLC method as described in Example 2.

Referring to the data in Tables 8 and 9, the Co-Lyophilized Combo Drug Product was characterized by amounts of the substances corresponding to HPLC peaks 1-12 that were less than the applicable drug product specification, indicating stabilization of the ceftolozane in the presence of about 484 mg of sodium chloride per 1,000 mg of ceftolozane active.

B. Stabilized Ceftolozane Compositions Lyophilized without Tazobactam

A composition comprising ceftolozane, and a stabilizing amount of sodium chloride was lyophilized, and then blended with a separately lyophilized composition of tazobactam. The stabilized ceftolozane composition was formed by lyophilizing an aqueous solution of the “CXA-101 for Injection Bulk” row of Table 10 (FIG. 10A), which was then blended with 5.4 g tazobactam free acid to form a pharmaceutical composition containing ceftolozane, sodium chloride and tazobactam components (“Blend Combo Drug Product”). Notably, this composition included about 481 mg of sodium chloride per 1,000 mg ceftolozane active provided as ceftolozane sulfate, and a weight ratio of about 2:1 between the CXA101 and tazobactam acid.

The results of a stability study of the Blend Combo Drug Product are shown in Table 11 (FIG. 10B) and Table 12 (FIG. 10C) as representative examples that summarize the results at 25° C./RH=60% and 40° C./RH=75% after one month (T1) and three months (T2). Samples were analyzed using a HPLC method as described in Example 2.

Referring to the data in tables 11 and 12, the Blend Combo Drug Product was characterized by amounts of the substances corresponding to HPLC peaks 1-12 that were less than the applicable drug product specification, indicating stabilization of the ceftolozane in the presence of about 481 mg of sodium chloride per 1,000 mg of ceftolozane active.

Example 5
Improvement in the Purity of Ceftolozane CXA-201 Pharmaceutical Compositions with Varying Amounts of Sodium Chloride

A stability study was carried out at 60° c. as described in Example 3. The sodium chloride content in the CXA-201 compositions is described in Table 13, FIG. 11. The HPLC data at 60° C. are summarized in Table 14-17, FIGS. 12-15.

	Number	Date	Country
	61793007	Mar 2013	US
	61792092	Mar 2013	US

CEFTOLOZANE PHARMACEUTICAL COMPOSITIONS

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