Formulation of a monobactam antibiotic

Field of the Invention

The present invention relates to stable pharmaceutical compositions comprising (1- (((Z)-(1-(2-aminothiazol-4-yl)-2-oxo-2-(((3S,4R)-2-oxo-4-((2 -oxooxazolidin-3-yl) methyl)-1-sulfoazetidin-3-yl)amino)ethylidene)amino)oxy)cycl opropane carboxylic acid of the Formula (I), their use in medicine and method of preparing said pharmaceutical compositions.

Background

Over the past several decades, the frequency of antimicrobial resistances and their association with serious infectious diseases have increased at alarming rates. The enhanced prevalence of resistances among nosocomial pathogens is particularly disconcerting. Of the over 2 million (hospital-acquired) infections occurring each year in the United States, 50 to 60% are caused by antibiotic-resistant strains of bacteria. The high rate of resistance to commonly used antibiotics increases the morbidity, mortality, and costs associated with nosocomial infections. In the United States, nosocomial infections are thought to contribute to or cause more than 77,000 deaths per year and cost approximately $5 to $10 billion annually.

Resistance of Gram-negative bacteria against antibiotics may be caused by extended-spectrum beta-lactamases (ESBLs), serine carbapenemases (KPCs) and metallo-beta-lactamases (for example NDM-1 ) in Klebsiella pneumoniae, Escherichia coli, and Proteus mirabilis, high-level third-generation cephalosporin (AmpC) beta-lactamase resistance among Enterobacter species and Citrobacter freundii, and multidrug-resistance genes observed in Pseudomonas, Acinetobacter, and Stenotrophomonas. The problem of antibacterial resistance is aggravated by the existence of bacterial strains resistant to multiple antibiotics. For example, Klebsiella pneumonia harboring an NDM-1 metallo-beta-lactamase carries frequently additional serine-beta-lactamases on the same plasmid that carries the NDM-1 .

Thus, there is a need for new antibiotics that are effective against existing drug- resistant microbes, or are less susceptible to development of new bacterial resistance. A new class of monobactam antibiotics is described in WO2015/148379, the content of which is herein incorporated by reference. In Example 22, the preparation of 1 -(((Z)-(1 -(2-aminothiazol-4-yl)-2-oxo-2-(((3S,4R)-2-oxo-4-((2-oxo- oxazolidin-3-yl)methyl)-1-sulfoazetidin-3-yl)amino)ethyliden e)amino)oxy)cyclo- propanecarboxylic acid (in the following compound (I)) is described.

Pharmaceutical compositions of compound (I) are disclosed in WO 2017/050218, the content of which is herein incorporated by reference. These compositions comprise large amounts of excipients such as sucrose for stabilizing the active agent compound (I). This makes preparing, handling and confectioning of these compositions cost-intensive, tedious and time-consuming. It would thus be beneficial to develop novel pharmaceutical compositions of compound (I).

It was an object of the present invention to developing stable pharmaceutical compositions of compound (I) that do not suffer from some or all of these disadvantages.

Summary of the invention

The present invention is directed to new solid and liquid pharmaceutical compositions of compound (I) and kits containing such compositions, which can be prepared and used in a shorter time, and in an improved, economic and simplified fashion.

A first aspect of the present invention relates to a solid pharmaceutical composition essentially consisting of:

- an active agent, which is (1-(((Z)-(1-(2-aminothiazol-4-yl)-2-oxo-2-(((3S,4R)-2-oxo- 4-((2-oxooxazolidin-3-yl)methyl)-1-sulfoazetidin-3-yl)amino) ethylidene)amino)oxy)cyclopropane carboxylic acid) of the Formula (I): including any tautomeric species, solvate or hydrate thereof; and

- a pH modifier.

A further aspect of the invention relates to a solid pharmaceutical composition essentially consisting of:

- an active agent, which is compound (I), including any tautomeric species, solvate or hydrate thereof; and

- L-arginine, a carbonate or any combination thereof.

A further aspect of the invention relates to a liquid composition essentially consisting of:

- an active agent, which is compound (I), including any tautomeric species, solvate or hydrate thereof;

- a pH modifier, and

- a pharmaceutically acceptable aqueous liquid, wherein the pH is in the range from about pH 4.0 to about pH 6.0.

A further aspect of the invention relates to a liquid composition essentially consisting of:

- an active agent, which is compound (I), including any tautomeric species thereof;

- L-arginine, a carbonate or any combination thereof, and

- water, particularly water for injection, wherein the pH is in the range from about pH 4.0 to about pH 6.0. A further aspect of the invention relates to a kit comprising:

(i) a solid pharmaceutical composition as described above;

(ii) a pharmaceutically acceptable aqueous liquid for reconstitution, and

(iii) optionally instructions for use.

A further aspect of the invention relates to methods of preparing a solid pharmaceutical composition, a liquid pharmaceutical composition or a kit as described above.

A further aspect of the invention relates to the use of a solid pharmaceutical composition, a liquid pharmaceutical composition or a kit as described above in medicine, particularly for the treatment of a bacterial infection.

General definitions

The term tautomeric species of compound (I) relates to compounds which differ from the depicted compound in that one or more H ⁺ ions are located at positions different from those indicated in the respective formula. For example, an NFh group may be in a protonated form, i.e. as an NH ₃ ⁺ group, a CO ₂ H group may be in a deprotonated form, i.e. as a CO ₂ group, and/or an SO ₃ H group may be in a deprotonated form, i.e. as an SO ₃ group. In certain embodiments, compound (I) includes a plurality of different tautomeric species, which may be in an equilibrium with each other depending on the pH. The term tautomeric species also includes zwitterionic species comprising both a protonated group and a deprotonated group.

The term solvate refers to a molecular complex of a compound of the present invention (including pharmaceutically acceptable salts thereof) with one or more solvent molecules, i.e. organic solvent molecules or water molecules. The term hydrate specifically refers to a complex where the solvent molecule is water.

A preferred hydrate of compound (I) is the trihydrate wherein compound (I) is bound to three molecules H ₂ O per molecule. Suitable solvates and hydrates of compound (I) are described in WO 2017/050218, the content of which is herein incorporated by reference.

As used in this specification and the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly indicates otherwise.

Similarly, “comprise”, “comprises”, “comprising”, “include”, “includes” and “including” are interchangeable and not intended to be limiting.

Detailed description

The present invention provides new stable solid compositions comprising compound (I) as an active agent, which are suitable for pharmaceutical use. The compositions may only consist of compound (I), particularly in an anhydrous amorphous form, and a pH modifier, particularly L-arginine and/or sodium carbonate, and are free from further excipients, particularly carbohydrates such as sucrose. They may be prepared by lyophilization of highly concentrated aqueous solutions of compound (I). Before use, they are reconstituted with an aqueous liquid, particularly water, and then administered to a patient, particularly by injection or infusion.

The solid composition of the present invention comprises an active agent, which is compound (I) including any tautomeric species, solvate or hydrate thereof. In certain embodiments, the active agent is the trihydrate of compound (I). In certain embodiments, the active agent is an anhydrous form, particularly an anhydrous amorphous form of compound (I).

The solid composition of the present invention comprises a pH modifier, i.e. a compound, which is suitable to achieve a desired pH in an aqueous solution. Suitable pH modifiers include sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium hydroxide, amines such as Tris (tris(hydroxymethyl)aminomethane), and amino acids, particularly basic amino acids such as arginine, lysine, and histidine. In certain embodiments, the pH modifier is present in amount for adjusting a pH in the range from about pH 4.0 to about pH 6.0, and particularly for adjusting a pH of about 5.0, e.g. pH 5 +/- 0.5 or pH 5 +/- 0.25. in an aqueous solution. In certain embodiments, the pH modifier is selected from the group consisting of a basic amino acid, a carbonate and any combination thereof.

In a specific embodiment, the pH modifier is L-arginine. Preferably, the composition comprises the active agent and L-arginine in a molar ratio in the range of about 1.5:1 to about 2:1 .

In a further specific embodiment, the pH modifier is sodium carbonate. Preferably, the composition comprises the active agent and sodium carbonate in a molar ratio of about 0.7:1 to about 1 :1.

The solid composition of the present invention essentially consists of the active agent compound (I) and a pH modifier. Thus, the presence of substantial amounts of further constituents is excluded. For example, the solid composition contains about 10 wt-% or less, about 5 wt-% or less, about 2 w-% or less, or about 1 wt-% or less of further constituents.

In certain embodiments, the solid composition has a total water content of about 2 wt-% or less, of about 1 wt-% or less, or of about 0.5 wt-% or less.

In certain embodiments, the solid composition is essentially free from further excipients, e.g. essentially free from any carbohydrates and/or essentially free from sucrose.

In particular embodiments, the solid composition consists of the active agent and the pH modifier, i.e. it does not contain any other constituent possibly except residual water or solvent, e.g., in an amount of about 0.5 wt-% or less and/or unavoidable impurities.

In certain embodiments, the solid composition of the present invention is a lyophilized composition, i.e., a composition prepared by lyophilization. Alternatively, the solid composition may be a composition prepared by spray drying. The solid composition may be in present in any suitable container, particularly in a container suitable for pharmaceutical use, e.g., a vial. The container may be a sterilized container, e.g., a sterilized vial. Further, the container optionally includes a stopper, e.g. a sterilized stopper, and/or a cap, e.g., a sterilized cap. The container may be made of glass and/or plastic.

The container may have any suitable volume, e.g., from about 1 ml_ to about 1000 ml_. Typically, the volume of the container is from about 10 mL to about 200 ml_, e.g., about 50 mL.

The amount of the composition is adapted to its intended use, e.g. use as a pharmaceutical agent. Typically, the composition may comprise about 1 g to about 50 g of the active agent, particularly about 2 g to about 6 g of the active agent and more particularly about 3 g of the active agent.

The solid composition of the present invention is typically adapted for reconstitution with a pharmaceutically acceptable liquid, particularly for reconstitution with water, particularly with water for injection.

The solid composition of the present invention is stable, i.e. it can be stored under suitable conditions without unacceptable degradation of the active agent, e.g. at a temperature of up to about 2-8°C for a time of at least 24 months of for at least 36 months.

In certain embodiments, the active agent is stable in that (i) it shows an increase of about 3 % or less, particularly about 2% or less impurities (measured as area-% by UPLC) from begin of the storage (t = 0) when stored in solid form at 5°C for 6 months; (ii) it shows an increase of 5 % or less, particularly about 4% or less impurities (measured as area-% by UPLC) from begin of the storage (t = 0) when stored in solid form at 25°C and 60% relative humidity for 6 months; and/or (ii) it shows an increase of about 10% or less, particularly about 9% or less impurities (measured as area-% by UPLC) from begin of the storage (t = 0) when stored in solid form at 40°C and 75% relative humidity for 6 months. Typically, the amount of impurities at begin of the storage (t = 0) is about 98% or more (measured as area-% by UPLC).

A further aspect of the present invention is directed to a solid pharmaceutical composition essentially consisting of or consisting of:

- an active agent, which is compound (I), including any tautomeric species thereof, e.g. in an anhydrous amorphous form or in the form of a trihydrate; and

- L-arginine, a carbonate, particularly sodium carbonate or any combination thereof.

A further aspect of the invention is directed to a liquid composition essentially consisting of:

- an active agent, which is compound (I), including any tautomeric species thereof,

- a pH modifier, and

- a pharmaceutically acceptable aqueous liquid, wherein the pH is in the range from about pH 4.0 to about pH 6.0, particularly of about pH 5.0, e.g. pH 5 +/- 0.5 or pH 5 +/- 0.25.

In certain embodiments, the liquid composition consists of the active agent, the pH modifier, and a pharmaceutically acceptable aqueous liquid. The pharmaceutically acceptable aqueous liquid comprises at least about 50% (v/v) water, at least about 80% (v/v) water or at least about 95% (v/v) water. In certain embodiments, the pharmaceutically acceptable aqueous liquid is water, particularly water for injection.

In certain embodiments, the liquid composition is a solution, i.e. it comprises the active agent and the pH modifier in a dissolved state.

In certain embodiments, the liquid composition comprises compound (I) in an amount, which is in the range of about 20 mg/mL to about 150 mg/mL, and which is particularly about 60 mg/mL. The liquid composition can be obtained by combining the individual constituents, i.e. active agent, pH modifier, and pharmaceutically acceptable aqueous liquid in the required amounts and preparing a solution. From such a solution, a solid composition as described above may be prepared by drying, e.g., lyophilization.

Further, the liquid composition can be obtained by reconstituting a solid composition as described above with a pharmaceutically acceptable aqueous liquid. Such a composition is ready for administration to a patient.

A further aspect of the invention is directed to a liquid composition essentially consisting of or consisting of:

- an active agent, which is compound (I), including any tautomeric species thereof,

- L-arginine, a carbonate or any combination thereof, and

- water, particularly water for injection, wherein the pH is in the range from about pH 4.0 to about pH 6.0.

A further aspect of the invention is directed to a kit comprising:

(i) a solid composition as described above;

(ii) a pharmaceutically acceptable aqueous liquid for reconstitution as described above and

(iii) optionally instructions for use.

A further aspect of the invention is directed to a method for preparing a solid composition as described above, comprising the steps:

(a) preparing a solution essentially consisting of the active agent, the pH modifier and a solvent, particularly an aqueous solvent, more particularly water, e.g. water for injection;

(b) optionally adjusting the pH of the solution to a target pH in the range from about pH 4.5 to about pH 6.0, e.g. pH 5 +/- 0.5 or pH 5 +/- 0.25.;

(c) filtering;

(d) optionally portioning,

(e) removing the solvent, and

(f) optionally confectioning. Filtration step (c) may comprise at least one filtration, e.g. two filtrations with a 0.22 urn filter. Optional portioning step (d) may comprise portioning the solution into predetermined batch sizes .According to step (e), the solvent may be removed by lyophilization or by spray drying. Optional confectioning step (f) may comprise at least one of filling the solid composition into a sterilized vial, stoppering the vial using a sterilized stopper, and capping the vial using a sterilized cap. The method may further comprise a quality control step, wherein the active agent and optionally of at least one impurity is quantitatively determined by suitable analytical methods including chromatographic methods such as HPLC.

A further aspect of the invention is directed to a method for preparing a liquid composition as described above, comprising reconstituting a solid composition, or a kit with a pharmaceutically acceptable aqueous liquid, particularly water.

The solid composition, the liquid composition or the kit are suitable for use in medicine, e.g. for use in human medicine, particularly for the treatment of a bacterial infection, e.g. a Gram-negative bacterial infection in a subject in need thereof. The bacterial infection may be selected from a chronic urinary tract infection, particularly a Gram-negative urinary tract infection, including a complicated urinary tract infection, a bacterial pneumonia, particularly a Gram-negative pneumonia, including a hospital acquired and/or ventilator-associated pneumonia, and a bacterial intra abdominal infection, particularly a Gram-negative intra-abdominal infection, including a complicated intra-abdominal infection. The bacterium causing the Gram negative bacterial infection may be selected from Citrobacter, Enterobacter, Escherichia, Haemophilus, Klebsiella, Morganella, Moraxella, Pseudomonas, Proteus, Salmonella, Serratia, Shigella, and Neisseria bacteria.

For therapeutic application, the solid composition or the kit may be reconstituted with a pharmaceutically acceptable aqueous liquid, particularly water, and administered to a subject in need thereof, e.g., a human subject. The reconstituted composition may be administered by injection or infusion, particularly by intravenous infusion. Other suitable routes include intramuscular, subcutaneous, intrathecal, and intraocular administration. Typically, the reconstituted composition is administered within a time period of up to 24 hours after reconstitution.

For therapeutic application, the liquid composition may be administered to a subject in need thereof, e.g., a human subject, by injection or infusion. Typically, the liquid composition is administered within a time period of up to 24 hours after preparation.

A further aspect of the invention is directed to a method for treating a bacterial infection in a subject in need thereof comprising administering an effective amount of compound (I), comprising reconstituting a solid composition as described above with a pharmaceutically acceptable aqueous liquid, particularly water, and administering the reconstituted composition to the subject, e.g., a human subject.

A further aspect of the invention is directed to a method for treating a bacterial infection in a subject in need thereof comprising administering an effective amount of compound (I), comprising administering a liquid composition as described above to the subject, a human subject.

The subject in need of treatment may suffer from a bacterial infection, e.g., from a Gram-negative bacterial infection as described above.

General Methods

The following HPLC method can be used for the detection of compound (I) and other compounds as described herein.

1

Principle RP UHPLC method with ion pairing and UV detection

Reagents Grade/Source

Acetonitrile: gradient grade, e.g. Merck LiChrosolv No. 100030

Methanol: gradient grade, e.g. Merck LiChrosolv No. 106007

Water deionized, for HPLC

Tetrabutylammonium HPLC grade, e.g. Sigma-Aldrich No. 86853 hydrogen sulphate (TBAHS) Trifluoroacetic acid (TFA) HPLC grade, e.g. Sigma-Aldrich No. 302031

1 M Hydrochloric acid 1 M Sodium hydroxide reagent grade, e.g. Fluka No. 35328 reagent grade, e.g. Fluka No. 35256

Diluent

Mobile Phase A

Materials

Glassware Amber colored volumetric glass flasks and UHPLC vials

Equipment

Apparatus UHPLC system with gradient elution and UV detector, e.g. Agilent 1290 with UV detector or equivalent

Column Waters Acquity HSS T3 Length: 100 mm, internal diameter: 2.1 mm Particle size: 1 .8 pm

Chromatographic conditions

Gradient

Mobile Phase A Water + 0.05% TFA + 5 mM TBAHS

Mobile Phase B Methanol + 0.05% TFA + 5 mM TBAHS

Gradient table Time [min] Phase A [%] Phase B [%] CO 98 2 1.0 98 2 20.0 60 40 26.0 54 46 32.0 30 70 36.0 30 70 36.1 98 2 38.1 98 2

43.1 70 30

43.2 98 2

50.0 98 2

Flow rate 0.4 mL/min Detection UV 260 nm

Column Temperature 40° C Autosampler Temperature 5°C Injection volume 2.0 pL of the test and reference solutions, equivalent to a column loading of approximately 2.4 pg of compound (I) drug substance (free acid).

Retention time (RT) compound (I) about 10.3 min

Calculation Percent of related substance (%RS): where

PAT Peak area of individual peaks in the test solution

Zi UV response factor

(ratio of the detector signal of a given amount of drug substance as salt free form divided by the detector signal of the same amount of related substance i as salt free form)

For unknown compounds Z = 1 .0

Examples

Example 1

Preparation of formulations containing compound (I) and L-arginine

Procedure:

Different formulations of compound (I) (in the following “API’) and L-arginine at pH 4.0, 4.5, 5.0, 5.5 and 6.0 were prepared at ambient room conditions; The initial amount of arginine for each formulation was dissolved in 75 mL WFI (water for injection);

12.5 g API were added slowly into the arginine solution with stirring until a clear solution was obtained; ^■ 0.5 M arginine solution in WFI was added to the formulation solution until the target pH was reached;

^■ WFI was added to make up for a volume of 100 ml_;

^■ The solution was filtered through PES 0.2-micron syringe filter; ■ 2 ml_ from each formulation were transferred into 48 vials, which were closed by a stopper. *The API was added as a hydrate that contains 11.8% water. Therefore, the actual API concentration in the compounding solution is 125 x 0.882 = 110 mg/ml_.

^■ The sample vials were immediately loaded into the Freeze Dryer ■ The lyophilization was carried out according to the following protocol:

^■ When the secondary drying was determined to be complete, the freeze dryer was stopped;

^■ The vials were backfilled with nitrogen and stoppered.

^■ An HPLC impurity analysis was performed for all 5 formulations (before freeze drying):

Example 2 Preparation of formulations containing compound (I) and sodium carbonate

Procedure:

^■ Different formulations of compound (I) (in the following “API’) and sodium carbonate at pH 4.0, 4.5, 5.0, 5.5 and 6.0 were prepared at ambient room conditions;

^■ The initial amount of sodium carbonate for each formulation was dissolved in 75 ml_ WFI (water for injection);

^■ 12.5 g API were added slowly into the arginine solution with stirring until a clear solution was obtained;

^■ 0.25 M sodium carbonate in WFI was added to the formulation solution until the target pH was reached;

^■ WFI was added to make up for a volume of 100 ml_;

^■ The solution was filtered through PES 0.2-micron syringe filter;

^■ 2 ml_ from each formulation were transferred into 48 vials, which were closed by a stopper.

^* The API was added as a hydrate that contains 11 .8% water. Therefore, the actual API concentration in the compounding solution is 125 x 0.882 = 110 mg/ml_.

^■ The sample vials were immediately loaded into the Freeze Dryer ■ The lyophilization was carried out according to the following protocol:

An HPLC impurity analysis was performed for all 5 formulation (before freeze drying):

Example 3

Stability testing

The following tables list the stability data of the formulations batches Arg-1 to Arg-5 (Example 1) and SC- to SC-5 (Example 2) each containing an amount of 250 mg lyophilisate. Measurements were performed at begin of storage, after 1 month (1 M), after 3 months (3 M) and after 6 months (6 M). 3.1 Formulation Arg-1

Arg-1: Storage temperature 5°C

Arg-1: Storage temperature 25°C/60% relative humidity (RH)

Arg-1: Storage temperature 40°C/75% RH 3.2 Formulation Arg-2

Arg-2: Storage temperature 5°C

Arg-2: Storage temperature 25°C/60% RH

Arg-2: Storage temperature 40°C/75% RH 3.3 Formulation Arg-3

Arg-3: Storage temperature 5°C

Arg-3: Storage temperature 25°C/60% RH

Arg-3: Storage temperature 40°C/75% RH 3.4 Formulation Arg-4

Arg-4: Storage temperature 5°C

Arg-4: Storage temperature 25°C/60% RH

Arg-4: Storage temperature 40°C/75% RH 3.5 Formulation Arg-5

Arg-5: Storage temperature 5°C

Arg-5: Storage temperature 25°C/60% RH

Arg-5: Storage temperature 40°C/75% RH 3.6 Formulation SC-1

SC-1 : Storage temperature 5°C

SC-1 : Storage temperature 25°C/60% RH

SC-1: Storage temperature 40°C/75% RH 3.7 Formulation SC-2

SC-2: Storage temperature 5°C

SC-2: Storage temperature 25°C/60% RH

SC-2: Storage temperature 40°C/75% RH 3.8 Formulation SC-3

SC-3: Storage temperature 5°C

SC-3: Storage temperature 25°C/60% RH

SC-3: Storage temperature 40°C/75% RH 3.9 Formulation SC-4

SC-4: Storage temperature 5°C

SC-4: Storage temperature 25°C/60% RH

SC-4: Storage temperature 40°C/75% RH 3.10 Formulation SC-5

SC-5: Storage temperature 5°C

SC-5: Storage temperature 25°C/60% RH

SC-5: Storage temperature 25°C/60% RH