Antibacterial parenteral formulations and methods thereof

Abstract

The present disclosure provides formulations comprising a compound of Formula (I), or a salt thereof, with a solubilizing agent, a hydrotrope, and a pH adjusting agent. The present disclosure also provides lyophilized formulations and reconstituted formulations. The present disclosure further provides processes for preparing the formulations and methods thereof. # imgabs0 #

Description

Technical Field

[0001] The present disclosure relates to the field of pharmaceutical preparations and in particular to antibacterial preparations. More specifically, the present disclosure relates to preparations of compounds (S)-6-(5-(((2-(7-fluoro-1-methyl-2-oxo-1,2-dihydroquinolin-8-yl)ethyl)amino)methyl)-2-oxooxazolidin-3-yl)-2H-pyrazino[2,3-b][1,4]oxazine-3(4H)-one or salts thereof comprising formula (I) and processes for preparing the preparations. The present disclosure also relates to freeze-dried injectable pharmaceutical preparations showing improved solubility and storage stability.

[0002] Background Art

[0003] Globally, the increasing challenges posed by the rising incidence and spread of antimicrobial resistance (AMR) necessitate the need to intensify efforts and avert this imminent major health threat. Recent epidemics have only reiterated the urgent need for health sectors to be fully prepared and take appropriate countermeasures to avoid a large-scale global disaster. There is a need to discover and develop new broad-spectrum antibiotics that can address AMR and provide solutions for the most serious hospital-associated infections. In addition to hospital-based and community-based health risks, the potential unexpected danger of bioterrorism is also particularly worrying because it may lead to a serious major public health emergency with panic, large-scale illness and death. Therefore, there is an urgent need for safe, potent broad-spectrum antibacterial agents that are effective against all key and high-priority pathogens with a multidrug resistance background (WHO Antimicrobial Resistance. https: / / www.who.int / news-room / fact-sheets / detail / antimicrobial-resistance). In this view, compounds of formula (I) can be considered suitable for the development of pharmaceutical formulations for antibiotic purposes.

[0004] The compound of formula (I) is a fully synthetic oxazolidinone-based broad-spectrum antibacterial agent and is chemically known as (S)-6-(5-(((2-(7-fluoro-1-methyl-2-oxo-1,2-dihydroquinolin-8-yl)ethyl)amino)methyl)-2-oxooxazolidin-3-yl)-2H-pyrazino[2,3-b][1,4]oxazin-3(4H)-one (WO2018225097). The compounds of formula (I) as broad-spectrum antibiotics are new chemical entities (NCEs) with dual-target inhibition and potent activity against a broad spectrum of Gram-negative bacteria (including all key members of Enterobacterales and non-fermenting bacteria, such as Pseudomonas aeruginosa and Acinetobacter baumannii, etc.) and Gram-positive bacteria (such as Methicillin resistant Staphylococcus aureus (MRSA) and Vancomycin resistant Enterococci (VRE), etc.) (WO2018225097). Extensive MIC studies of nearly 10,000 clinical isolates showed no cross-resistance to all currently used antibiotics, including fluoroquinolones.

[0005] However, the compounds of formula (I) need to be developed as parenteral antibiotics to treat various bacterial infections and have good solubility in pharmaceutically acceptable solvents in the pH range of 3 to 8. In addition, the formulation or drug product should be stable in the formulated solvent and suitable for long-term storage.

[0006] Therefore, there is clearly still a need to develop pharmaceutically acceptable injectable drug formulations of formula (I) which can be prepared and stored as ready-to-use drugs and which meet the requirements regarding stability for longer storage periods. Summary of the invention

[0007] In one aspect of the present disclosure, there is provided a formulation comprising: a) a compound of formula (I) or a salt thereof;

[0008]

[0009] b) a solubilizing agent; c) a hydrotropic agent; and d) a pH adjusting agent.

[0010] In another aspect of the present disclosure, there is provided a lyophilized formulation comprising: a) a compound of formula (I) or a salt thereof;

[0011]

[0012] b) a solubilizing agent; c) a hydrotrope; and d) a pH adjusting agent.

[0013] In another aspect of the present disclosure, a reconstituted formulation is provided, comprising a lyophilized formulation as disclosed herein and a reconstitution agent and a diluent.

[0014] In yet another aspect of the present disclosure, a kit is provided, comprising: a) a first compartment comprising a formulation as disclosed herein; b) a second compartment comprising an enhancer; and c) optional accessories.

[0015] In another aspect of the present disclosure, there is provided a process for preparing a formulation as disclosed herein, the process comprising: a) contacting a solubilizing agent with a compound of formula (I) or a salt thereof in the presence of a second solvent under stirring at a temperature in the range of 18 to 40° C. to obtain a first solution; b) adding a hydrotrope to the first solution under stirring to obtain a second solution; and c) mixing a pH adjuster into the second solution and then adding a vehicle to obtain the formulation.

[0016] In further aspects of the present disclosure, there is provided a process for preparing a lyophilized formulation as disclosed herein, the process comprising: a) freezing the formulation as disclosed herein at a temperature ranging from 0 to -50°C, and then drying at a pressure ranging from 10 μbar to 1 bar to obtain a lyophilized formulation.

[0017] In another aspect of the present disclosure, there is provided a process for preparing a reconstituted formulation as disclosed herein, the process comprising contacting the lyophilized formulation with a diluent and then adding a reconstitution agent.

[0018] In another aspect of the present disclosure, a method of treating a bacterial infection is provided, the method comprising administering to a subject in need thereof an effective amount of a formulation as disclosed herein.

[0019] In further aspects of the present disclosure, provided are methods of treating or preventing a disease or disorder, the methods comprising administering to a subject in need thereof an effective amount of a formulation as disclosed herein.

[0020] These and other features, aspects and advantages of the subject matter will be better understood with reference to the following description and the appended claims. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify the key features or essential features of the subject matter claimed, nor is it intended to be used to limit the scope of the subject matter claimed. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Shown is an XRD (x-ray diffraction) diffraction pattern of a formate salt of a compound of formula (I) according to an embodiment of the present disclosure.

[0022] Figure 2 Shown is a DSC (differential scanning calorimetry) thermogram of a formate salt of a compound of formula (I) according to an embodiment of the present disclosure.

[0023] Figure 3 The pXRD (powder x-ray diffraction) diffraction pattern of the compound of formula (I) according to an embodiment of the present disclosure is shown.

[0024] Figure 4 Shown are CFU / g of thighs of mice infected with Acinetobacter baumannii ATCC 17978 and treated with formulations of the present disclosure or meropenem (200 mg / kg) or polymyxin B (25 mg / kg) q8h subcutaneously, according to embodiments of the present disclosure. DETAILED DESCRIPTION

[0025] Those skilled in the art will appreciate that the present disclosure may be subject to variations and modifications other than those specifically described. It should be understood that the present disclosure includes all such variations and modifications. The present disclosure also includes all such steps, features, compositions and compounds mentioned or indicated in this specification, either individually or collectively, and any and all combinations of any one or more such steps or features.

[0026] definition

[0027] For convenience, before further describing the present disclosure, certain terms and examples used in the specification are collected here. Those skilled in the art should read and understand these definitions based on the remainder of the present disclosure. The terms used herein have meanings recognized and known by those skilled in the art, however, for convenience and completeness, specific terms and their meanings are listed below.

[0028] The articles “a,” “an,” and “the” are used to refer to one or to more than one (ie, to at least one) of the grammatical object of the article.

[0029] The terms "comprise" and "comprising" are used in an inclusive, open sense, meaning that additional elements may be included. Throughout the specification, unless the context requires otherwise, the word "comprise" and variations such as "comprises" and "comprising" will be understood to imply the inclusion of a stated element or step or group of elements or steps, but not the exclusion of any other element or step or group of elements or steps.

[0030] The term "including" is used to mean "including but not limited to." "Including" and "including but not limited to" are used interchangeably.

[0031] As used herein, the term "w / w" refers to the weight of a component relative to the total weight of the composition. As used herein, the term w / v refers to the weight of a component relative to the total volume of the composition. In addition, when water is used in the formulation, then w / w and w / v should be used interchangeably.

[0032] The term "solubilizing agent" as used herein refers to a substance that helps dissolve otherwise insoluble substances in water. The solubilizing agents of the present disclosure include but are not limited to lactic acid, ascorbic acid, acetic acid, propionic acid, succinic acid, gluconic acid, benzoic acid, tartaric acid, glutaric acid, malic acid and fumaric acid. The terms "solubilizing agent" and "solubilizers" are used interchangeably. The solubilizing agent includes but is not limited to lactic acid, ascorbic acid, acetic acid, propionic acid, succinic acid, gluconic acid, benzoic acid, tartaric acid, glutaric acid, malic acid, fumaric acid or a combination thereof.

[0033] The term "hydrotrope" as used herein refers to a chemical or substance added to an incompatible mixture of two or more different substances. A hydrotrope is added to increase the solubility of the solvent in the mixture. Hydrotropes include, but are not limited to, niacinamide, sodium benzoate, sodium citrate, sodium acetate, or a combination thereof.

[0034] The term "pH adjusting agent" as used herein refers to a substance that can be added to a solution or mixture to provide a desired pH. In the present disclosure, pH adjusting agents include, but are not limited to, potassium hydroxide, sodium hydroxide, L-arginine, histidine, glycine, sodium bicarbonate, or a combination thereof. The terms "pH modifier" and "pH adjusting agents" are used interchangeably.

[0035] The term "vehicle" as used herein refers to a substance or liquid used as a medium for dispersing, suspending or dissolving a material. In the present disclosure, the term "vehicle" refers to water, which is added to the formulation to make the formulation injectable and easy to use.

[0036] The term "diluent" as used herein refers to a substance used to make the formulation easy to flow or pump. In the present disclosure, diluents include but are not limited to dextrose, sorbitol, sodium bicarbonate, glucose, mannitol, sucrose or sodium chloride.

[0037] The term "reconstitution agent" as used herein refers to a substance that is added to a dry substance and converts the dry substance into a fluid form. In the present disclosure, a reconstitution agent refers to a substance that is added to a lyophilized preparation and makes the preparation easy to use, preferably as an injectable preparation. The reconstitution agent of the present disclosure includes but is not limited to water. A reconstitution agent is added to a lyophilized preparation to obtain a reconstituted preparation.

[0038] The term "lyophilized preparation" as used herein refers to a dry preparation, wherein the prepared preparation is subjected to a continuous process such as heating and freezing to obtain a lyophilized preparation. Lyophilized preparations enhance the stability of the active ingredients in the preparation and provide long-term storage of the drug product. Lyophilized preparations are also referred to as lyophilized cakes, wherein the prepared preparation is subjected to a process selected from heating, freezing, drying or a combination thereof, which is performed in a sequential manner to obtain a lyophilized preparation. The process for obtaining a lyophilized preparation is also referred to as a lyophilization cycle (lyocycle).

[0039] As used herein, the term "therapeutic agent" refers to a substance or pharmaceutical material capable of imparting a therapeutic effect, such as diagnosis, prevention, cure, alleviation or treatment of a disease, disorder, condition or infection.

[0040] Once a term is described, the same meaning applies throughout the disclosure.

[0041] As discussed in the background art, compounds of formula (I) have the potential to treat a variety of bacterial infections. In particular, compounds of formula (I) are capable of treating a wide variety of clinical indications caused by both Gram-positive and Gram-negative bacterial species, such as complicated and uncomplicated urinary tract infections (cUTI, e.g., pyelonephritis, cystitis), intra-abdominal infections (cIAI), bloodstream infections, hospital-acquired bacterial pneumonia (HABP, nosocomial pneumonia) and ventilator-associated bacterial pneumonia (VABP), community-acquired bacterial pneumonia (CABP), cystic fibrosis secondary infection (CFI), skin and soft tissue infection (SSTI), endocarditis, meningitis, dysentery and diarrhea, typhoid fever, Clostridium difficile-associated colitis and diarrhea (CDAD), Helicobacter pylori-associated peptic ulcer. It also has the potential to treat bacterial sexually transmitted diseases caused by Chlamydia trachomatis, gonorrhea caused by Neisseria gonorrhoeae, and syphilis caused by Treponema pallidum, as well as bioterrorism-related bacterial diseases caused by anthrax (Bacillus anthracis), plague (Yersinia pestis), tularemia (Francisella tularensis), glanders (Burkholderia mallei), melioidosis (Burkholderia pseudomallei), and Q fever (Coxiella burnetii).

[0042] However, the solubility of the compound of formula (I) or its salt is limited and therefore bioavailability may be limited. It should be noted that the compound of formula (I) contains an amine group and therefore it can easily form salts with acids, including hydrochlorides, formate, acetates, etc. However, the salt form of the compound of formula (I) has a very limited solubility (2-4 mg / ml) in water and needs to be further improved in a suitable pharmaceutical solvent with a pH compatible with parenteral administration for human use with a 20-50 mg / ml strength. Therefore, the present disclosure provides a preparation comprising a compound of formula (I) and various components such as a solubilizing agent, a hydrotrope and a pH regulator. In addition, the preparation needs to be stable for a long time and should also maintain the stability of the preparation in the form of injection. The preparation of the present disclosure uses a combination of acids to obtain better solubility and further uses a pH regulator to balance the pH caused by the combination of acids. In addition, the preparation of the present disclosure uses a suitable hydrotrope to maintain the maximum solubility of the compound of formula (I) in the preparation. In addition, the present disclosure provides a lyophilized formulation, a reconstituted formulation, and a process for preparing the same.

[0043] In an embodiment of the present disclosure, a formulation is provided, comprising: a) a compound of formula (I) or a salt thereof;

[0044]

[0045] b) a solubilizing agent; c) a hydrotrope; and d) a pH adjusting agent.

[0046] In an embodiment of the present disclosure, a formulation as disclosed herein is provided, wherein the formulation comprises a compound of formula (I) or a salt thereof in the weight range of 1 to 30% (w / w); a solubilizing agent in the weight range of 2 to 55% (w / w); a hydrotrope in the weight range of 1 to 30% (w / w); and a pH adjusting agent in the weight percentage range of 0.5 to 30% (w / w).

[0047] In another embodiment of the present disclosure, a formulation as disclosed herein is provided, wherein the formulation comprises a compound of formula (I) or a salt thereof in the weight range of 2 to 20% (w / w); a solubilizing agent in the weight range of 5 to 35% (w / w); a hydrotrope in the weight range of 1 to 20% (w / w); and a pH adjusting agent in the weight percentage range of 0.5 to 20% (w / w).

[0048] In an embodiment of the present disclosure, a formulation as disclosed herein is provided, wherein the solubilizing agent is selected from lactic acid, ascorbic acid, acetic acid, propionic acid, succinic acid, gluconic acid, benzoic acid, tartaric acid, glutaric acid, malic acid, fumaric acid or a combination thereof; the pH adjusting agent is selected from potassium hydroxide, sodium hydroxide, l-arginine, histidine, glycine, sodium bicarbonate or a combination thereof; the hydrotropic agent is selected from nicotinamide, sodium benzoate, sodium citrate or sodium acetate; and the salt of the compound of formula (I) is selected from formate, methanesulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, acetate, propionate, fumarate, maleate, tartrate, succinate, glycolate, glutamate, aspartate, hydrochloride, hydrobromide or sulfate. In another embodiment of the present disclosure, wherein the solubilizing agent is selected from lactic acid, ascorbic acid, acetic acid or a combination thereof; the pH adjusting agent is selected from potassium hydroxide, l-arginine or a combination thereof; and the hydrotropic agent is nicotinamide or sodium benzoate.

[0049] In an embodiment of the present disclosure, there is provided a formulation as disclosed herein, wherein the solubilizing agent is a combination of lactic acid and ascorbic acid; the pH adjusting agent is l-arginine; the hydrotrope is nicotinamide; and the salt of the compound of formula (I) is a formate salt.

[0050] In an embodiment of the present disclosure, there is provided a formulation as disclosed herein, wherein the formulation is stable at a temperature ranging from -70°C to 10°C.

[0051] In an embodiment of the present disclosure, there is provided a formulation as disclosed herein, wherein the pH of the formulation is in the range of 2 to 6. In another embodiment of the present disclosure, the pH of the formulation is in the range of 3 to 5.

[0052] In an embodiment of the present disclosure, there is provided a formulation as disclosed herein, wherein the formulation comprises a vehicle; and the vehicle is water.

[0053] In an embodiment of the present disclosure, there is provided a formulation as disclosed herein, wherein the formulation is capable of killing or inhibiting the growth of a microorganism, and the microorganism is selected from bacteria, viruses, fungi or protozoa.

[0054] In an embodiment of the present disclosure, there is provided a formulation as disclosed herein, wherein the formulation is an anti-infective agent against microorganisms, particularly bacteria; and the bacteria are selected from Gram-negative bacteria, Gram-positive bacteria, or a combination thereof.

[0055] In an embodiment of the present disclosure, a formulation as disclosed herein is provided, wherein the microorganism is selected from Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Enterobacter cloacae, Citrobacter spp., Proteus spp., Serratia marcescens, Salmonella species, Morganella morganii, Klebsiella oxytoca, Klebsiella aerogenes, Providencia spp., Neisseria gonorrhoea, Mycoplasma spp., Campylobacter sp., Fusobacterium spp., Bacteriodes fragilis. fragilis), Prevotellas p., Shigella sp., Helicobacter pylori, Ureaplasma spp., Burkholderia gladioli, Burkholderia multivorans, Pandorea apista, Burkholderia cepacia species, Burkholderia pseudomallei, Burkholderia glanders, Stenotrophomonas maltophilia, Achromobacter spp.), Ralstonia picketii, Legionella pneumophila, Clostridium difficile, Staphylococcus aureus, Coagulase-negatives Staphylococcus, Streptococcus pyogenes, Streptococcus pneumoniae, Enterococcus faecium, Enterococcus faecalis, Bacillus anthracis, Yersinia pestis, and Francisella tularensis.

[0056] In an embodiment of the present disclosure, there is provided an agent as disclosed herein, wherein the agent inhibits an enzyme selected from bacterial gyrase, topoisomerase IV or a combination thereof.

[0057] In an embodiment of the present disclosure, there is provided a formulation as disclosed herein, wherein the formulation further comprises an additive, a therapeutic agent or a combination thereof.

[0058] In an embodiment of the present disclosure, there is provided a formulation as disclosed herein, wherein the additive is selected from sorbitol, dextrose, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium citrate, sodium bicarbonate, sodium chloride, or a combination thereof.

[0059] In an embodiment of the present disclosure, a formulation is provided, which comprises: a) a compound of formula (I) or a salt thereof; b) a solubilizing agent; c) a hydrotrope; d) a pH adjusting agent; and e) a vehicle.

[0060] In an embodiment of the present disclosure, a formulation is provided, which comprises: a) a compound of formula (I) or a salt thereof; b) a solubilizing agent; c) a hydrotrope; d) a pH adjusting agent; e) a solvent; and f) an additive.

[0061] In an embodiment of the present disclosure, a formulation is provided, which comprises: a) a compound of formula (I) or a salt thereof; b) a solubilizing agent; c) a hydrotrope; d) a pH adjusting agent; e) a solvent; f) an additive; and g) a therapeutic agent.

[0062] In an embodiment of the present disclosure, a formulation is provided, which comprises: a) a compound of formula (I) or a salt thereof; b) a solubilizing agent selected from lactic acid, ascorbic acid, acetic acid, propionic acid, succinic acid, gluconic acid, benzoic acid, tartaric acid, glutaric acid, malic acid, fumaric acid, or a combination thereof; c) a hydrotrope selected from niacinamide, sodium benzoate, sodium citrate, and sodium acetate; and d) a pH adjusting agent selected from potassium hydroxide, sodium hydroxide, l-arginine, histidine, glycine, sodium bicarbonate, or a combination thereof.

[0063] In an embodiment of the present disclosure, a formulation is provided, which comprises: a) a compound of formula (I) or a salt thereof; b) a solubilizing agent selected from lactic acid, ascorbic acid, acetic acid, propionic acid, succinic acid, gluconic acid, benzoic acid, tartaric acid, glutaric acid, malic acid, fumaric acid or a combination thereof; c) a hydrotrope selected from niacinamide, sodium benzoate, sodium citrate and sodium acetate; d) a pH adjusting agent selected from potassium hydroxide, sodium hydroxide, l-arginine, histidine, glycine, sodium bicarbonate or a combination thereof; and e) water as a solvent.

[0064] In an embodiment of the present disclosure, a formulation is provided comprising: a) a formate salt of a compound of formula (I); b) a solubilizing agent that is a combination of ascorbic acid and lactic acid; c) niacinamide; and d) l-arginine.

[0065] In an embodiment of the present disclosure, a lyophilized formulation is provided, which comprises: a) a compound of formula (I) or a salt thereof; b) a solubilizing agent; c) a hydrotrope; and d) a pH adjuster.

[0066] In an embodiment of the present disclosure, a reconstituted formulation is provided, comprising: a lyophilized formulation comprising a) a compound of formula (I) or a salt thereof, b) a solubilizing agent, c) a hydrotrope, and d) a pH adjuster, as well as a reconstitution agent and a diluent.

[0067] In an embodiment of the present disclosure, a reconstituted formulation as disclosed herein is provided, wherein the reconstituted agent is water; the diluent is selected from dextrose, sorbitol, sodium bicarbonate, glucose, mannitol, sucrose or sodium chloride; and the weight range of the diluent in the first solvent is 0.9-25% (w / v). In another embodiment, the weight range of the diluent in the first solvent is 1-10% (w / v). In another embodiment, the weight range of the diluent in the first solvent is 2-10% (w / v).

[0068] In an embodiment of the present disclosure, there is provided a reconstituted formulation as disclosed herein, the first solvent being water.

[0069] In an embodiment of the present disclosure, a reconstituted formulation is provided, which comprises: i) a lyophilized formulation comprising a) a compound of formula (I) or a salt thereof, b) a solubilizing agent, c) a hydrotrope, d) a pH adjuster; ii) a reconstitution agent; and iii) a diluent.

[0070] In an embodiment of the present disclosure, a reconstituted formulation is provided, which comprises: a) a compound of formula (I) or a salt thereof; b) a solubilizing agent selected from lactic acid, ascorbic acid, acetic acid, propionic acid, succinic acid, gluconic acid, benzoic acid, tartaric acid, glutaric acid, malic acid, fumaric acid or a combination thereof; c) a hydrotrope selected from niacinamide, sodium benzoate, sodium citrate and sodium acetate; d) a pH adjusting agent selected from potassium hydroxide, l-arginine, histidine, glycine, sodium bicarbonate or a combination thereof; e) water as a reconstitution agent; and f) a diluent selected from dextrose, sorbitol, sodium bicarbonate, glucose, mannitol, sucrose or sodium chloride.

[0071] In an embodiment of the present disclosure, a reconstituted formulation is provided, comprising: a) a formate salt of a compound of formula (I); b) a solubilizing agent which is a combination of ascorbic acid and lactic acid; c) niacinamide; d) l-arginine; e) water as a reconstitution agent; and f) dextrose as a diluent.

[0072] In an embodiment of the present disclosure, a kit is provided, comprising: a) a first compartment comprising a formulation as disclosed herein; b) a second compartment comprising an enhancer; and c) optional accessories.

[0073] In an embodiment of the present disclosure, a kit is provided, comprising: a) a first compartment comprising a formulation as disclosed herein; b) a second compartment comprising water for injection; and c) optional accessories selected from a diluent bag, an infusion tube, a needle, a connector, or a combination thereof.

[0074] In an embodiment of the present disclosure, a process for preparing a formulation is provided, the process comprising: a) contacting a solubilizing agent with a compound of formula (I) or a salt thereof in the presence of a second solvent under stirring at a temperature in the range of 18 to 40° C. to obtain a first solution; b) adding a hydrotrope to the first solution under stirring to obtain a second solution; and c) mixing a pH adjuster into the second solution and then adding a solvent to obtain a formulation.

[0075] In an embodiment of the present disclosure, a process for preparing a formulation as disclosed herein is provided, wherein the solubilizing agent is selected from ascorbic acid, lactic acid, acetic acid, succinic acid or a combination thereof; and two or more solubilizing agents are mixed prior to contacting with the compound of formula (I).

[0076] In an embodiment of the present disclosure, a process for preparing a formulation as disclosed herein is provided, wherein the pH of the first solution is in the range of 2 to 3; and the pH of the second solution is in the range of 2.5 to 4.

[0077] In an embodiment of the present disclosure, a process for preparing a formulation as disclosed herein is provided, wherein the formulation is filtered after adding the solvent.

[0078] In an embodiment of the present disclosure, there is provided a process for preparing a formulation as disclosed herein, wherein the second solvent is water; and the vehicle is water.

[0079] In an embodiment of the present disclosure, a process for preparing a lyophilized formulation is provided, the process comprising: a) freezing the formulation as disclosed herein at a temperature ranging from 0 to -50°C, and then drying at a pressure ranging from 10 μbar to 1 bar to obtain a lyophilized formulation.

[0080] In an embodiment of the present disclosure, a process for preparing a lyophilized formulation as disclosed herein is provided, wherein freezing is performed for a time period ranging from 30 minutes to 30 hours; and drying is performed for a time period ranging from 20 minutes to 150 hours. In another embodiment of the present disclosure, freezing is performed for a time period ranging from 1 hour to 25 hours; and drying is performed for a time period ranging from 1 hour to 140 hours. In another embodiment of the present disclosure, wherein freezing is performed for a time period ranging from 10 hours to 25 hours; and drying is performed for a time period ranging from 50 hours to 130 hours.

[0081] In an embodiment of the present disclosure, a process for preparing a reconstituted formulation is provided, the process comprising contacting a lyophilized formulation with a diluent and then adding a reconstitution agent.

[0082] In an embodiment of the present disclosure, there is provided a process for preparing a reconstituted formulation as disclosed herein, wherein the process is performed for a time period ranging from 60 seconds to 10 minutes.

[0083] In an embodiment of the present disclosure, there is provided a process for preparing a reconstituted formulation as disclosed herein, wherein the reconstituted formulation is stable at a temperature in the range of 20 to 35°C for a period of time in the range of 20 to 30 hours.

[0084] In an embodiment of the present disclosure, there is provided a process for preparing a reconstituted formulation as disclosed herein, wherein the pH of the reconstituted formulation is in the range of 3 to 5 and the osmotic pressure is in the range of 300-500 mOs-mol / kg.

[0085] In an embodiment of the present disclosure, there is provided use of a formulation or a lyophilized formulation or a reconstituted formulation as disclosed herein.

[0086] In an embodiment of the present disclosure, there is provided a use of a kit as disclosed herein.

[0087] In an embodiment of the present disclosure, there is provided use of a formulation or a lyophilized formulation or a reconstituted formulation as disclosed herein for the manufacture of a medicament.

[0088] In an embodiment of the present disclosure, a method of treating a bacterial infection is provided, the method comprising administering to a subject in need thereof an effective amount of a formulation as disclosed herein.

[0089] In an embodiment of the present disclosure, a method of treating a bacterial infection is provided, the method comprising administering to a subject in need thereof an effective amount of a lyophilized formulation as disclosed herein.

[0090] In an embodiment of the present disclosure, a method of treating a bacterial infection is provided, the method comprising administering to a subject in need thereof an effective amount of a reconstituted formulation as disclosed herein.

[0091] In an embodiment of the present disclosure, a method of treating a bacterial infection as disclosed herein is provided, wherein the bacterial infection is caused by bacteria, viruses, fungi, or protozoa.

[0092] In an embodiment of the present disclosure, a method of treating or preventing a disease or disorder is provided, the method comprising administering to a subject in need thereof an effective amount of a formulation as disclosed herein.

[0093] In an embodiment of the present disclosure, a method of treating or preventing a disease or disorder is provided, the method comprising administering to a subject in need thereof an effective amount of a lyophilized formulation as disclosed herein.

[0094] In an embodiment of the present disclosure, there is provided a method of treating or preventing a disease or disorder, the method comprising administering to a subject in need thereof an effective amount of a reconstituted formulation as disclosed herein.

[0095] In embodiments of the present disclosure, methods of treating or preventing a disease or condition as disclosed herein are provided, wherein the disease or condition is mediated by a Gram-positive bacterial species, a Gram-negative bacterial species, or a combination thereof.

[0096] In an embodiment of the present disclosure, a method of treating or preventing a disease or condition as disclosed herein is provided, wherein the disease or condition is selected from complicated and uncomplicated urinary tract infection (cUTI, e.g., pyelonephritis, cystitis), intra-abdominal infection (cIAI), bloodstream infection, hospital-acquired bacterial pneumonia (HABP, nosocomial pneumonia), ventilator-associated bacterial pneumonia (VABP), community-acquired bacterial pneumonia (CABP), cystic fibrosis secondary infection (CFI), skin and soft tissue infection (SSTI), endocardial difficile-associated colitis and diarrhea (CDAD), Helicobacter pylori-associated peptic ulcer, sexually transmitted diseases caused by Chlamydia trachomatis, gonorrhea caused by Neisseria gonorrhoeae, syphilis caused by Treponema pallidum, or bioterrorism-related bacterial diseases caused by anthrax (Bacillus anthracis), plague (Yersinia pestis), tularemia (Franciscoides tularensis), glanders (Burkholderia mallei), melioidosis (Burkholderia pseudomallei), and Q fever (Coxiella burnetii).

[0097] Although the subject matter has been described in considerable detail with reference to certain embodiments of the subject matter, other embodiments are possible. Therefore, the spirit and scope of the present disclosure should not be limited to the description of the embodiments contained herein.

[0098] Example

[0099] The following examples provide details on the synthesis, activity and use of the compounds and formulations of the present disclosure. It should be understood that the following is representative only, and the present disclosure is not limited to the details set forth in these examples.

[0100] Abbreviations

[0101] XRD - X-ray diffraction

[0102] DMSO-Dimethyl sulfoxide

[0103] CFU-colony forming unit MIC-minimum inhibitory concentration pXRD-powder X-ray diffraction PPt-precipitate

[0104] PBS - Phosphate Buffered Saline NMT - not more than

[0105] WFI-water for injection mL-milliliter

[0106] mg - milligrams

[0107] g-gram

[0108] Qs - enough

[0109] ℃-Celsius

[0110] Hrs-hours

[0111] RH - relative humidity

[0112] FD-Formulation Development

[0113] API-Active Pharmaceutical Ingredient Ppt-Precipitation

[0114] PVP K-12-Polyvinylpyrrolidone HPβ-CD-Hydroxypropylβ-cyclodextrin NaOH-Sodium hydroxide N-Equivalent (Normality)

[0115] RT - Room temperature

[0116] USP-United States PharmacopoeiaMin-minute

[0117] mT-mitorr

[0118] R / H-Ramp / Hold IV-Intravenous

[0119] w / w - weight / weight

[0120] Sec-seconds

[0121] mM - millimole

[0122] Methods and Materials

[0123] For the purposes of this disclosure, the following materials were purchased and used without further purification.

[0124]

[0125] Powder X-ray diffraction analysis was performed using a Bruker Model D2 PHASER instrument. Thermal analysis and differential scanning calorimetry were performed using a Perkin Elmer instrument Model No: DSC8000.

[0126] Example 1

[0127] Compounds of formula (I)

[0128] The compound of formula (I) is prepared by the process defined in WO20188225097 and obtained as a crystalline formate salt (S)-6-(5-(((2-(7-fluoro-1-methyl-2-oxo-1,2-dihydroquinolin-8-yl)ethyl)amino)methyl)-2-oxooxazolidin-3-yl)-2H-pyrazino[2,3-b][1,4]oxazine-3(4H)-one formate having a single polymorphic characteristic.

[0129]

[0130] The prepared formate salt of the compound of formula (I) was subjected to powder x-ray diffraction analysis and the characteristic single polymorph XRD diffraction pattern is shown in Figure 1 The clear pattern of the diffraction pattern confirmed the formation of a single polymorph with crystalline characteristics. Figure 2 Depicted is a Differential Scanning Calorimetry (DSC) thermogram of the formate salt of the compound of formula (I). It can be inferred that the compound of formula (I) is crystalline in nature, as indicated by the tall and sharp peaks of the thermogram.

[0131] Example 2

[0132] Antimicrobial activity assay

[0133] The compounds of formula (I) exhibit very potent antibacterial activity against various bacterial species including Gram-negative and Gram-positive bacteria, aerobic and anaerobic bacteria, and biothreat bacterial pathogens. Bacterial susceptibility testing procedures for various bacterial species are described herein.

[0134] MIC broth microdilution method

[0135] The following MIC broth microdilution method is used for all bacterial strains except N. gonorrhoeae, H. pylori, and anaerobic bacteria:

[0136] Antimicrobials were prepared according to the CLSI susceptibility test standard (Clinical and Laboratory Standards Institute CLSI. 2018. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically. 11th edition. CLSI standard M07). A solution of 2× final concentration of antimicrobial or combination was prepared in a 50 mL test tube by diluting in CA-MHB (or buffered yeast extract alpha ketoglutarate broth for Legionella pneumophila) and manually transferred to a 96-well master block. Then 50 μL of each well was transferred from the master block to a 96-well plate. The microtiter plates were stored frozen at -80°C until the test day, for up to 6 months, and frozen / thawed no more than once. Approximately 1×10 was prepared by diluting a 0.5McFarland suspension 100 times in CA-MHB with TES. 6 CFU / mL of bacterial inoculum. Thaw the antibacterial plate wells at room temperature and add 50 μL of inoculum to each well to obtain approximately 5×10 5The final density of CFU / mL and the desired test concentration of the antimicrobial agent. The test plates were incubated according to CLSI guidelines and read visually. The MIC value corresponds to the first well with no visible growth.

[0137] Agar Dilution Method for Neisseria gonorrhoeae / Helicobacter pylori

[0138] The compound of formula (I) was dissolved in DMSO at 3200 μg / mL. A solution of 100× test range concentration was prepared in DMSO by serial dilution, and then diluted 100 times in GC agar medium (for Neisseria gonorrhoeae) or Mueller-Hinton agar supplemented with 5.0% aged (≥2 weeks) sheep blood (for Helicobacter pylori) and poured into a culture dish (final volume = 40 mL). MIC tests were performed by agar dilution according to CLSI sensitivity test standards (CLSI.2021.PerformanceStandards for Antimicrobial Susceptibility Testing. 31st edition.CLSI supplementM100). Bacterial inoculum was prepared by direct colony suspension method. The bacterial suspension was adjusted to 0.5McFarland in 0.9% NaCl. 200 μL of bacterial suspension was placed in a well of a 96-well plate, and 1 μL of each suspension (equivalent to approximately 1×10 4 CFU) were deposited on the agar surface. The plates were incubated according to CLSI guidelines (CLSI. 2018. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically. 11th edition. CLSI standard M07).

[0139] Agar dilution method for antimicrobial testing of anaerobic bacteria (Bacteroides, Fusobacterium, Prevotella and Clostridium)

[0140] The compound of formula (I) or its salt, preferably the formate of the compound of formula (I) is dissolved in DMSO with 3200 μg / mL. A solution of 100 × test range concentration is prepared in DMSO by serial dilution, then diluted 100 times in Brucella agar supplemented with 5 μg / mL hemin, 1 μg / mL vitamin K1 and 5.0% lysed sheep blood, and the culture medium is poured into a culture dish (final volume = 40 mL). Before determining the MIC value, a Bactron 600 anaerobic chamber containing 5% CO2, 5% H2 and 90% N2 atmosphere is used to incubate the solution at 35-37 ° C for 42-48 hours (CLSI.2021.Performance Standards for Antimicrobial Susceptibility Testing. 31st edition.CLSI supplement M100).

[0141] Minimum inhibitory concentration (MIC) is the lowest concentration of active ingredient or drug that helps prevent bacterial growth. 50 (minimum inhibitory concentration that prevents 50% bacterial growth of the tested species) and MIC 90 (Minimum inhibitory concentration that prevents 90% bacterial growth of the species tested) and are listed below in Table 1. The formate salt of the compound of formula (I) exhibited potent antibacterial activity against Gram-positive, Gram-negative, anaerobic and biothreat bacterial pathogens (Table 2).

[0142] Table 1: MICs of the formate salt of the compound of formula (I) against clinical isolates of Gram-negative and Gram-positive species 90 result

[0143]

[0144]

[0145] Table 2: MIC of formate salts of compounds of formula (I) against biological threat pathogens 90 result

[0146]

[0147] *N indicates the number of bacterial strains tested in a particular species.

[0148] Example 3

[0149] Salt screening

[0150] Salt screening of compounds of formula (I) was evaluated to select suitable salts for developing formulations with improved solubility in water. The solid-state properties of compounds of formula (I) were investigated using pXRD diffraction patterns. Figure 3 ) The free amine of the compound of formula (I) was found to be amorphous in nature as X-ray diffraction showed a halo diffraction pattern in the range of 2-40° (2θ).

[0151] Choice of counterion

[0152] It was found that the free amine compound of formula (I) was only soluble in a mixture of dichloromethane and methanol (1:1). Therefore, the same solvent system was used during the salt screening experiment using the slow solvent evaporation method. Due to the weak basicity (pKa ~ 6.81) of the compound of formula (I), acidic counterions were used to screen its salt-forming tendency. In theory, counterions with a pKa value at least 2 units smaller than the basic pKa value of the compound of formula (I) are considered for salt formation. Table 3 shows a list of selected counterions used for salt screening.

[0153] Table 3

[0154] Counter ions pKa p-Toluenesulfonic acid -1.34 Methanesulfonic acid -2.6 Camphorsulfonic acid 1.5 Maleic acid 2.0,6.26 Malonic acid 2.8,5.67 Malic Acid 3.4,5.11 Succinic acid 4.19,5.57 Fumaric Acid 3.03,4.44 Anhydrous Oxalic Acid 1.25,4.23 Benzenesulfonic acid -2.8 Ethylsulfonic acid 1.68 ascorbic acid 4.17&11.6 Citric Acid 3.13,4.76&6.40 lactic acid 3.86 Formic acid 3.75

[0155] A common solvent approach in which the drug and counterion are soluble was optimized and salt screening was performed. Qualitative solubility of the counterion was determined in water as solvent and a stoichiometric ratio of 1:1.125 of drug and counterion was used to prepare solutions during salt screening experiments.

[0156] The compound of formula (I) (100 mg) was dissolved in an appropriate amount of an organic solvent system comprising a mixture of dichloromethane and methanol (1: 1) and the desired amount of counterions dissolved in methanol were added. Then, it was heated at 50° C. to 60° C. for a few minutes and allowed to equilibrate to room temperature to observe whether any salt formation occurred due to crystallization or precipitation from the solution. The results of the salt screening are summarized in Table 4 below.

[0157] Table 4 Feasibility test of free amine salts of compounds of formula (I)

[0158] Acidic counter ions observe HCl Solid Formation p-Toluenesulfonic acid A solution was observed initially. A solid formed after evaporation of the solvent. Methanesulfonic acid A solution was observed initially. A solid formed after evaporation of the solvent. Camphorsulfonic acid Solid Formation Maleic acid PPt. Formed after counterion addition. Solid formed. Malonic acid PPt. Formed after counterion addition. Solid formed. Fumaric Acid PPt. Formed after counterion addition. Solid formed. tartaric acid PPt. Formed after counterion addition. Solid formed. Succinic acid PPt. formed after counterion addition. Solid formed. However, gel formation was observed Anhydrous Oxalic Acid PPt. Formed after counterion addition. Solid formed. Malic Acid PPt. Formed after counterion addition. Solid formed. Citric Acid PPt. Formed after counterion addition. Solid formed. ascorbic acid Remains solid for 1-2 days, then shows deliquesce Formic acid Solid Formation

[0159] *Ppt-precipitate

[0160] The counterions of the crystalline salts generated from the above experiments were further considered for scale-up experiments (200 mg scale).The solubility of the selected crystalline salts was then evaluated in various aqueous media to examine the solubility enhancement of the salts of the compound of formula (I).

[0161] The compound of formula (I) as free amine (200 mg) was weighed into a round bottom flask and dissolved in sufficient amount of dichloromethane and methanol (1:1) mixture with the aid of ultrasonic treatment and slight heating on a water bath at about 40-50° C. Then, the required amount of acidic counterion (if solid) was weighed and dissolved in methanol or measured (if liquid counterion) and then transferred to the above contents in the round bottom flask and kept stirring overnight to observe the precipitated solid and collect the solid after complete evaporation of the solvent.

[0162] Solubility of selected salts of compounds of formula (I)

[0163] The qualitative solubility of the salt of the compound of formula (I) prepared by evaluation in various aqueous media is to select the best counterion based on the solubility enhancement compared with formate. The salt of the compound of formula (I) of 2 mg equivalent is weighed in a transparent glass vial. Add thereto the solvent listed in 20 μ L of the following table 5 and carry out ultrasonic treatment, vortex and observe whether to dissolve. If not dissolved, then add the solvent of further increment as given in table 5. Carry out ultrasonic treatment and vortex to dissolve the compound when each increment.

[0164] Table 5: Qualitative solubility of various screened salts of the compound of formula (I)

[0165]

[0166] Inference from salt screening:

[0167] In order to strengthen the solubility of the compound of formula (I), salt screening is carried out to meet the high solubility requirement of the higher dosage required for human administration. Various salts are screened using acidic counterions, such as sulfonate, mesylate, succinate, toluenesulfonate, fumarate, formates, maleate, malate, oxalate, lactate, glucuronate and HCl. In the case of maleate, citrate and oxalate tests, preliminary feasibility tests have produced amorphous salts. Even in the case of lactate and ascorbate, amorphous solids are also formed. However, from Table 5, it can be determined that the formates of the compound of formula (I) only show crystallinity and increased solubility.

[0168] The formate salt of the compound of formula (I) gave a 10 mg / mL solution in both 10% w / v aqueous ascorbic acid and 30% v / v aqueous lactic acid, while the HCl salt and malate salt gave clear solutions at a concentration of 5 mg / mL in 30% aqueous lactic acid.

[0169] Determination of the saturated solubility of the formate salt of the compound of formula (I)

[0170] The saturated solubility of the formate salt of the compound of formula (I) was measured at 25°C ± 2°C in various solvents, acids, additives as listed in Table 6A.

[0171] Water for injection (WFI) and various solvents (5mL or 5.0g) are placed in 10mL transparent glass bottles. After adding the compound formate of formula (I) of appropriate amount and closing with stopper in each bottle, these glass bottles are installed in orbital shaking water bath. Shaking is carried out for 72 hours, and temperature is maintained at about 25 ± 2 ℃. Then, the drug concentration determination of the obtained test sample in each relevant solvent is evaluated at 24 hours, 48 ​​hours and 72 hours.

[0172] Table 6A: Saturation solubility results of the formate salt of the compound of formula (I)

[0173]

[0174]

[0175] The formate salt of the compound of formula (I) exhibits very limited solubility in water and different water-soluble solvents, however, the maximum solubility is exhibited in individual acids such as ascorbic acid, acetic acid and lactic acid. The combination of acid with hydroxypropyl β-cyclodextrin (HPβ-CD) and polyvinylpyrrolidone (PVP) K-12 shows solubility enhancement for the formate salt of the compound of formula (I). It can be observed that the use of ascorbic acid, lactic acid, glacial acetic acid in combination with HPβCD and PVP K-12 leads to the maximum solubility of the formate salt of the compound of formula (I).

[0176] pH solution stability study

[0177] pH solution stability was performed at 25°C ± 2°C in the following buffers listed in Table 6B (50 mM acetate buffer for pH 3.0, pH 4.0, pH 5.0 and 50 mM phosphate buffer for pH 6.0).

[0178] In order to determine the pH solution stability of the compound formate of formula (I) in different pH ranges, the corresponding buffer is prepared by dissolving the buffer salt in WFI. The compound formate of the formula (I) of the weighed amount is added to each buffer system and dissolved. The volume is supplemented with WFI. The 5mL sample is loaded into a 5mL USPI type vial. The vial is stoppered and sealed. These vials are maintained at 25°C for stability studies. Blank buffer / placebo samples are also placed with the active samples. The properties (description), pH, assay content (assay), and related substances of the samples are evaluated at 24 hours, 48 ​​hours, and 72 hours.

[0179] Table 6B: Compositions used for pH solution stability studies of formate salts of compounds of formula (I) (batch size: 50 mL each)

[0180]

[0181] NA: Not applicable

[0182] Table 6C: Results of pH solution stability studies of formulations in Table 6B

[0183]

[0184] I: Initial, *transparent light yellow solution

[0185] Table 6D: Results of pH solution stability studies of formulations in Table 6B

[0186]

[0187]

[0188] I: Initial, * transparent light yellow solution; ** off-white suspension

[0189] The formulation was found to be a transparent light yellow solution at pH 3.0, 4.0 and 5.0, but it was an off-white suspension at pH 6.0. After 24 hours at 25°C, the content decreased in all pH ranges. The decrease was between 0.4% and 3.5%. This shows that the compound formate of formula (I) has a solution stability of up to 24 hours at pH 3.0-pH 5.0. The related substance (RS) data show that the impurity level increases after 48 hours at 25°C. Compared with pH 3.0, 4.0 and 5.0, the increase of RS at pH 6.0 is significant. Therefore, for formulation development, the target pH range will be between pH 3.0 and 5.0.

[0190] Example 4

[0191] Compatibility study

[0192] The medicine herein is referred to as active ingredient, which is a compound of formula (I) or its salt. Compatibility studies of medicine and formulation components are carried out to determine the suitable formulation components for realizing acceptable impurity profiles. In order to achieve this, compatibility and stability studies of 1 month were carried out at 25°C / 60%RH and 40°C / 75%RH in various components such as buffer, solvent, surfactant mixture in solution / suspension state. Mixture stability was carried out for 1 month at 25°C / 60%RH and 40°C / 75%RH for all these batches.

[0193] Table 7 below provides details of the container closure system used in the present disclosure.

[0194] Table 7: Details of container closure system

[0195]

[0196] Formulations for compatibility studies

[0197] Various formulations F1 to F5 were prepared using the components as described in Table 8 below. In addition, stability and solubility studies were performed on each formulation and the results are listed in Tables 9, 10 and 11.

[0198] Table 8: Formulations used for drug compatibility studies

[0199]

[0200] *X indicates the absence of that particular component

[0201] Table 9: Results of drug-component compatibility studies of F1 and F2

[0202]

[0203] *Pale yellow suspension; #Light brick red solution with brown particles ##Yellow suspension with green pellets

[0204] Table 10: Results of drug-excipient compatibility studies for F3 and F4

[0205]

[0206]

[0207] *Pale yellow suspension; ##Yellow suspension with green lumps

[0208] Table 1: Results of drug-component compatibility study of F5

[0209]

[0210] *Light yellow suspension; @@@@Dense brick red suspension

[0211] In the drug-component compatibility study, significant changes in appearance, assay content and related substances (RS) were observed under both conditions, i.e., at 25°C ± 2°C / 60 ± 5% RH and 40°C ± 2°C / 75 ± 5% RH, as shown in Tables 9, 10 and 11. The changes in appearance (color of the solution / suspension) were observed under both conditions mainly due to discoloration of ascorbic acid, since the placebo (formulation without the compound of formula (I)) sample containing ascorbic acid also showed discoloration. Compared with the assay content of the batches placed with the individual components, the decrease in the assay content was greater in the case of the batches placed with the formulation, except for the batch with L-arginine (F4).

[0212] The impurity profile in the batches with individual components (F1, F2, F3 and F4) was found to be lower compared to the batch manufactured as a formulation (F5).

[0213] The solubility of the compound of formula (I) or its salt is enhanced

[0214] The solubility of the formate salt of the compound of formula (I) in water was limited and was further improved to achieve the target formulation specifications. This was achieved by using a combination of acids such as ascorbic acid, lactic acid and glacial acetic acid.

[0215] The formate salt of the compound of formula (I) is unstable in the higher pH range, i.e. in the neutral to alkaline pH range (pH 5.0 to 10.0) and is very stable at acidic pH (<5). Therefore, the desired target pH range for formulations comprising the formate salt of the compound of formula (I) is between pH 3.0 and pH 5.0. The formate salt of the compound of formula (I) is highly unstable in the liquid state (solution / suspension); therefore, the pharmaceutical formulation is manufactured as a lyophilized pharmaceutical formulation to obtain a stable product.

[0216] It is believed that different prototype formulations use a combination of acids to improve the solubility of the formate salt of the compound of formula (I). Vortex and mix at 35°C-40°C to dissolve the formate salt of the compound of formula (I). Once the solution temperature reaches 25°C-30°C, record all observations.

[0217] Among the solubilizers studied, the formate salt of the compound of formula (I) was most soluble in acidic conditions and a combination of acids such as ascorbic acid, lactic acid, and acetic acid. In addition, with the addition of solubilizers such as hydroxypropyl β-CD and polyvinylpyrrolidone K-12, the solubility was enhanced to greater than 25 mg / mL.

[0218] Table 12: Results of solubility enhancement of the formate salt of the compound of formula (I).

[0219]

[0220]

[0221]

[0222] NR: Not Recorded

[0223] Use of L-arginine as a pH adjuster and evaluation of formulations

[0224] L-arginine was used as a pH adjuster and the formulation was evaluated to maintain the formulation pH between 3.0 and 4.0. Two different batches of 10 mL of compound formate of formula (I) (SF19000421 and SF19000435) were prepared. The prepared formulations for evaluation are shown in Table 13. Formulation 35A, in addition to the components of formulation 21A, also included l-arginine.

[0225] Table 13: Formulations used for pH solution stability studies of the formate salt of the compound of formula (I) (21A)

[0226]

[0227]

[0228] Table 14: Observations of pH solution stability studies of compounds of formula (I)

[0229]

[0230] * Transparent light yellow solution

[0231] Table 14 shows the solution stability at the beginning of the preparation and after 18-20 hours. It can be seen that L-arginine shows a stable pH and the appearance of the solution remains transparent for up to 18 hours. Therefore, it is found that L-arginine is suitable for use as a pH adjuster for the preparation of the compound of formula (I).

[0232] Evaluation of pH adjuster and hydrotrope combination

[0233] Formulations with pH modifiers (pH adjusters) and hydrotropes (in the presence and absence) were evaluated, with the pH of the formulations ranging from 3.5 to 4.0. Table 15 provides formulations of solution stability of the compound formate salt of formula (I) at different pH ranges and their corresponding stability studies are shown in Table 16.

[0234] Table 15: Preparation of solution stability of formate salt of compound of formula (I) at different pH ranges

[0235]

[0236]

[0237] % expressed as w / v

[0238] Table 16: Observation results of solution stability of formate salt of compound of formula (I) in different pH ranges

[0239]

[0240] *Transparent light yellow solution; **Light yellow ppt;

[0241] ***Dark yellow to brownish yellow ppt. #Immediately light yellow ppt.

[0242] Among the pH adjusters and solubilizers evaluated, the combination of L-arginine and nicotinamide showed better stability (appearance of solution) up to 3 to 4 hours and up to 20 hours (Formulation 465B). Therefore, l-arginine and nicotinamide were used as pH adjusters and hydrotropes, respectively. It can be further observed that the pH adjuster should be taken in a specific weight % range, i.e., in the range of 0.5-30%, and when taken outside of said range, resulted in precipitation of the formulation.

[0243] Evaluation of the concentration of acid (solubilizing agent) that dissolves the formate salt of the compound of formula (I).

[0244] Formulations prepared using different acid concentrations were evaluated to obtain the desired concentrations of solubilization of the formate salt of the compound of formula (I) as shown in Table 17, and their observations are listed in Table 18 below.

[0245] Table 3: Formulations of different acid concentrations for dissolving the formate salt of the compound of formula (I).

[0246]

[0247]

[0248] Table 18: Observations on optimization of acid concentration to dissolve the formate salt of the compound of formula (I)

[0249]

[0250] *Transparent light yellow solution; **Hazy light yellow solution;

[0251] ***Turbid yellow solution; #Light yellow ppt; ##Transparent solution with a small amount of particles.

[0252] The concentration of glacial acetic acid and lactic acid has a significant effect on the solubilization of the compound of formula (I). Formulations 470B and 470G show the solubility of the compound formate of formula (I) at a concentration of 25 mg / mL. In the remaining formulations of Table 17, the compound formate of formula (I) cannot be completely dissolved. Formulation 470A is considered to be a control sample. Therefore, the minimum acid concentration as shown in Table 19 is required to dissolve the compound formate of formula (I) of different concentrations.

[0253] Table 19: Composition of the minimum acid concentration required to dissolve the formate salt of the compound of formula (I).

[0254]

[0255] Based on the improved solubility observed using organic acids, the formulation was further evaluated by using a combination of ascorbic acid and lactic acid to increase solubility, reducing the overall acid strength, with a desired pH range between 4.00 and 4.50.

[0256] Evaluation of the solubility of the formate salt of the compound of formula (I) (without acetic acid)

[0257] Formulations comprising the formate salt of the compound of formula (I) were prepared with different concentrations of lactic acid as a solubilizing agent and without acetic acid as shown in Table 20. The appearance, pH of the prepared formulations were observed and recorded in Table 21.

[0258] Table 4: Formulations used for solubility studies of the formate salt of the compound of formula (I) without acetic acid

[0259]

[0260] Table 21: Observations from solubility studies of the formate salt of the compound of formula (I) without acetic acid

[0261]

[0262]

[0263] As can be seen from Table 21 above, the compound of formula (I) was dissolved at a concentration of 20 mg / mL using a combination of ascorbic acid and lactic acid, and the pH of the solution was found to be in the range of pH 4.5 to 5.3. Therefore, it is inferred that acetic acid may optionally be used as a solubilizing agent.

[0264] Example 5

[0265] Evaluation of the lyophilization cycle of the formate salt of the compound of formula (I)

[0266] The lyophilization cycles of the formulations of the present disclosure were evaluated based on the lyophilization cycles of the formulations with two acid combinations, namely ascorbic acid and lactic acid. The formulations were prepared using a batch size of 100 mL each and the formulations are shown in Table 22.

[0267] Table 22: Formulations for lyophilization cycle development of compounds of formula (I)

[0268]

[0269] The formulations of Table 22 were prepared as described herein. Ascorbic acid (solubilizer) was dissolved in water for injection (~70% batch size, 25°C-30°C), then lactic acid (solubilizer) was added and dissolved by stirring for 5 minutes to obtain a solution. The solution was heated to 35°C-40°C. The compound formate of formula (I) was added to the solution and dissolved by stirring for 2 to 3 hours. The solution was then placed to cool to room temperature under continuous stirring. Nicotinamide (hydrotropes) was added to the first solution and dissolved by stirring for 10 minutes to obtain a second solution. L-arginine was added to the above solution and dissolved by stirring for 10 minutes to obtain a formulation. Mannitol (according to the corresponding batch) was then added and dissolved under stirring for 10 minutes. The solution volume was made up to the batch size using water for injection (25°C-30°C) (solvent) and mixed for 15 minutes.

[0270] The solution was filtered through a 0.2μ PES filter (47 mm) using a vacuum filtration assembly. 25.0 mL of the filtered solution was loaded into a 50 mL clear USPI type tubular vial, the vial was half-stoppered and loaded for lyophilization. Lyophilization cycles were performed as shown in Table 23 below.

[0271] Table 5: Lyophilization cycle of batch No 529 of Table 22

[0272]

[0273]

[0274] R: temperature rise and temperature drop; H: maintain

[0275] Total freeze drying cycle time: 132.3 hours

[0276] After lyophilization the vacuum was broken and the vials were stoppered, unloaded from the lyophilizer, and sealed. The moisture content observed in the two batches was 529A: 5.52% w / w and 529B: 7.65% w / w and the observed results for the lyophilized samples are shown in Table 24.

[0277] Table 24: Results of freeze drying of batch No 529

[0278]

[0279] The above freeze-drying cycle batch provided acceptable cake appearance, reconstitution time and was further used to produce prototype stability batches. Based on the moisture content, secondary drying needs to be performed at a higher temperature, i.e., 25°C-30°C. Therefore, further freeze-drying cycle optimization was performed for prototype batch development.

[0280] Preliminary dilution studies of the formate salt of the compound of formula (I) as an injection sample

[0281] The lyophilized formulations prepared as above were evaluated for dilution studies and 5% dextrose (GS, diluent) solution, sterile water for injection were used for dilution studies. The batches of formulations used were B.No. 529A and 529B.

[0282] The lyophilized formulation was reconstituted with sterile water for injection (reconstitution agent) and the reconstituted formulation was further diluted with different diluents, wherein the ratio of the compound of formula (I) formate salt to the diluent was 1: 5. As shown in Tables 25 and 26, the appearance, pH and osmotic pressure of the reconstituted and diluted samples were evaluated.

[0283] Table 6: Results of reconstituted formulations

[0284]

[0285] Table 7: Results of the dilution study

[0286]

[0287] *Transparent slightly yellowish solution

[0288] The reconstituted solution is a clear light yellow solution and is stable for up to 6 hours at room temperature. The product was diluted with 5% glucose solution and showed an osmotic pressure between 380-410 mOsmol / Kg, which is within the acceptable range for low volume parenteral administration. However, the sample diluted with sterile water for injection showed a hypotonic solution (140-170 mOsmol / Kg).

[0289] Example 6

[0290] Prototype stability batches of parenteral dosage forms

[0291] Prototype formulation batches for stability studies were prepared as detailed in Table 27 below, each batch size was 800 mL, and lyophilized formulations were prepared as shown in Table 28. Stability studies were performed on the lyophilized formulations and the results were recorded for each batch as shown in Tables 29-32.

[0292] Table 27: Compositions used for prototype stability of formulations

[0293]

[0294]

[0295] Table 8: Lyophilization cycle formulation of prototype batch of compound formula (1) formate salt drug product

[0296]

[0297]

[0298] R: temperature rise and temperature drop; H: maintain total freeze drying cycle time: 134.33 hours

[0299] Table 99: Stability data of prototype batch_SF19000586 IUN: initial unfiltered solution; IF: initial filtered solution

[0300]

[0301]

[0302] *White cake on top and red cake on bottom; **Transparent yellow solution @Red yellow color cake completely shrunk / collapsed into a ball; @@Yellow suspension; ***Pale yellow solution with a small amount of grayish brown to black particles $: Reddish yellow color cake slightly collapsed; #: Sample was kept for 5 minutes after adding WFI to wet the cake. Then shaken vigorously. (Time required to reconstitute the sample)

[0303] Table 30: Stability data of prototype batch_SF19000601

[0304]

[0305]

[0306] IUN: Initial Unfiltered Solution; IF: Initial Filtered Solution

[0307] *White cake on top and red cake on bottom;

[0308] **Transparent yellow solution

[0309] #Red and yellow cakes completely shrink / collapse into balls

[0310] ##Yellow to orange suspension

[0311] $Off-white to light yellow cake, with slightly collapsed structure

[0312] @: Samples were kept for 5 minutes after addition of WFI to wet the cake. Then shaken vigorously. (Time required to reconstitute the sample (min)).

[0313] Table 10: Stability data of prototype batch_SF19000634

[0314]

[0315]

[0316] IUN: Initial unfiltered solution; IF: Initial filtered solution * Off-white to light yellow cake; ** Transparent slightly light yellow solution

[0317] ##Off-white to light red cake, with slight collapse; ***Yellow to orange suspension

[0318] $Off-white to light yellow cake, with a slightly collapsed structure

[0319]

[0320] Table 11: Stability data of prototype batch_SF19000649

[0321] IUN: initial unfiltered solution; IF: initial filtered solution#off-white to light yellow cake; *transparent slightly light yellow solution;

[0322] ##Off-white to light yellow cake, with collapse / meltback; **Transparent yellow solution

[0323] ###Light yellow to red cake; ***Yellow suspension

[0324] All batches of the formulation of the compound formate of formula (I) showed remelting / collapse of the cake at 40°C±2°C / 75%RH±5%RH after 6 months, and at 25°C±2°C / 60%RH±5%RH, a slightly collapsed cake structure with a change in appearance compared to the initial was observed in all batches. In all batches, the lyophilized cake after 6 months at 40°C±2°C / 75%RH±5%RH and 25°C±2°C / 60%RH±5%RH showed difficulty in reconstitution, resulting in a yellow to orange suspension.

[0325] The stability of the lyophilized formulations after 6 months at 2°C-8°C (all batches) showed a cake structure similar to the initial cake and no difficulty in the reconstitution of the lyophilized formulations. All batches of formulations did not show pH changes in the stored formulations under all stability conditions. For formulations stored at 40°C ± 2°C / 75% RH ± 5% RH and 25°C ± 2°C / 60% RH ± 5% RH, a decrease in the assay content and an increase in impurities were observed, while for batches of formulations stored at 2°C-8°C, no significant differences were observed in the assay content and impurity profiles. Mannitol had no significant effect on the stability of the drug product.

[0326] Therefore, a lyophilized formulation of the formate salt of the compound of formula (I) for injection was evaluated under different pH conditions, namely at pH 3.50, 4.00 and 4.50 and storage conditions of 2°C-8°C selected for pH range studies. The developed formulation was further slightly adjusted for process optimization.

[0327] Study on process optimization of injection preparations

[0328] The formate salt of the compound of formula (I) was not dissolved at room temperature even after stirring for 3 hours, but heating to 40°C-50°C dissolved the compound of formula (I) in the preparation with the solvent. This resulted in a significant effect of temperature and stirring time on the related substances of the preparation in the unfiltered sample and the filtered sample, especially in the sample prepared by stirring at 50°C for 3 hours. Therefore, the process was further optimized to dissolve the formate salt of the compound of formula (I) at a lower temperature.

[0329] Effect of initial volume of WFI on solubilization of the formate salt of the compound of formula (I)

[0330] The formulations of the present disclosure were prepared by varying the process temperature of the formulation preparation and its effect on the solubilization of the formate salt of the compound of formula (I) was studied. A batch size of 50 mL each was used.

[0331] Table 12: Effect of process temperature on the solubilization of the formate salt of the compound of formula (I)

[0332]

[0333]

[0334] % expressed as w / v

[0335] Ascorbic acid and lactic acid were dissolved in 20% of the total batch size of water for injection (25°C-30°C). Then, the compound formate of formula (I) was added to the above solution and kept stirring under the corresponding temperature conditions as shown in Table 33. Nicotinamide was added to the above solution at room temperature (RT) and dissolved by stirring. L-arginine was added to the above solution and dissolved by stirring. The volume was made up to 50 mL with WFI and kept stirring at room temperature. The sample was kept at room temperature for physical observation.

[0336] Table 13: Observation results of process optimization for solubilization of compound of formula (I)

[0337]

[0338] * Transparent light yellow solution ** Transparent light yellow solution with dust-like particles

[0339] From Table 34 above, it can be understood that the formate salt of the compound of formula (I) can be dissolved at 25°C (room temperature, RT) and 45°C using a smaller amount of WFI (ie 20% of the total batch size) in the initial stage of the manufacturing process.

[0340] The solutions produced using this process remain stable at room temperature (5 to 18 hours) and demonstrate that the heating step in the manufacturing process can be eliminated when dissolving the formate salt of the compound of formula (I) in a given solvent formulation as shown herein.

[0341] To determine the glass transition temperature, differential scanning calorimetry (DSC) thermogram analysis was performed. In the DSC thermogram, no glass transition temperature was observed. An exothermic peak at -20°C to -25°C was observed in both thermograms; this was due to the crystallization of water in the sample and the endothermic peak at 0°C indicated ice melting phenomenon.

[0342] During cooling (second time) to -70°C, a peak was observed at -32°C to -36°C. However, this peak may be due to recrystallization of the frozen matrix. Using DSC analysis, it is challenging to detect the glass transition temperature of the formulation, because the formulation is composed of acid and different salts, which may have very low glass transition temperatures. Further research is needed to determine the glass transition temperature or collapse temperature.

[0343] Lyophilization cycle development for formulations

[0344] Based on the lyophilized formulation developed herein, further optimization was performed. An annealing step was introduced during the freezing phase to form larger ice crystals. The preparation process was as follows, i.e., the formate salt of the compound of formula (I) was dissolved at room temperature and the lyophilization cycle was further developed to obtain a higher solid content.

[0345] Table 35: Formulations developed for the lyophilization cycle of the formate salt of the compound of formula (I)

[0346]

[0347] The preparations as shown in Table 35 were prepared by the process defined herein. Ascorbic acid (solubilizer) was dissolved in water for injection (second solvent, batch size of ~20%, 25°C-30°C), then lactic acid (solubilizer) was added under stirring for 5 minutes. Then, the compound of formula (I) as formate was added to the above solution and dissolved by stirring at room temperature for 3 hours to obtain the first solution. Nicotinamide (hydrotropic agent) was then added to the first solution and dissolved for 10 minutes to obtain the second solution, followed by L-arginine (pH regulator) added under stirring for 10 minutes to obtain the preparation. The volume of the preparation was made up to the batch size using WFI (25°C-30°C) and mixed for 15 minutes. The solution was filtered through a 0.2μPES filter (47mm) using a vacuum filtration assembly. Each preparation with a batch size of 300mL was prepared.

[0348] Table 36 below shows the process followed by the freeze drying cycle of the compound formate of formula (I). As shown in Table 36 below, the preparation is frozen in a temperature range of -50°C to 0°C, preferably in a range of -45°C to 0°C. After freezing, the preparation is subjected to additional freezing and drying to obtain a freeze-dried sample. Drying is carried out in a pressure range of 10 μbar to 1 bar. After freeze drying, the vacuum is broken with nitrogen. The vial is stoppered, unloaded from the freeze dryer, and sealed. The appearance and characteristics of the freeze-dried preparation are analyzed as shown in Table 37.

[0349] Table 36: Lyophilization cycle of the formate salt of the compound of formula (I) - Batch No. SF19000829

[0350]

[0351]

[0352] R: heating and cooling; H: maintaining

[0353] Total freeze drying cycle time: 97.08 hours

[0354] Table 37: Results for the lyophilized formulation of the formate salt drug product of the compound of formula (I) - Batch No. SF19000829

[0355]

[0356] The annealing step during freezing helps to improve the cake structure and provide a complete cake. The freeze-dried cake is easily reconstituted with water for injection within 2 minutes to form a transparent light yellow solution. No pH change / pH movement of the reconstituted solution due to freeze-drying is observed. Then, the freeze-drying cycle is slightly adjusted at the end of the primary drying step, with a freeze-drying cycle of 96 hours as the target, which provides similar freeze-dried cake structure, reconstruction time and uses the same freeze-drying cycle for further development, with a higher solid content in the freeze-dried cake.

[0357] Diluent compatibility study of formulations

[0358] Diluent compatibility studies were performed using 5% dextrose injection at 1.0 mg / mL, 3.33 mg / mL, and 10.0 mg / mL strengths after dilution.

[0359] Table 38

[0360]

[0361]

[0362] The lyophilized formulations as shown in Table 38 were prepared according to the following process. Water for injection (WFI) was purged with nitrogen for 1 hour. 20% of the total batch size of water for injection (RT) purged with nitrogen was collected in a glass beaker. A weighed amount of ascorbic acid was added thereto and stirred for 5 minutes at room temperature using a magnetic stirrer. The solution was transparent and was a light yellow solution. A weighed amount of lactic acid was added to the above solution under stirring at room temperature. The solution was transparent and was still a light yellow solution. A weighed amount of the compound formate of formula (I) was added to the above solution and stirred for 120 minutes at room temperature. The solution now became translucent and was a dark yellow solution with a small amount of dust-like particles. 40% of the total batch size of WFI purged with nitrogen was added to the above solution and stirred for 10 minutes at room temperature. A translucent yellow solution with some dust-like particles was observed. A weighed amount of nicotinamide was added to the above solution and stirred for 15 minutes at room temperature. A translucent dark yellow solution with some dust-like particles was observed. The L-arginine of the weighed amount is added to the above solution and kept stirring at room temperature for 15 minutes. The solution becomes a translucent dark yellow solution with some dust-like particles. The volume of the above step solution is made up to the required batch size by using WFI (room temperature) purged with nitrogen and stirred at room temperature for 15 minutes. (Appearance: transparent to translucent, dark yellow solution, with some dust (external) particles). The above solution is filtered through a 0.2 μ PES (polyethersulfone) membrane filter, and the appearance (after filtration) is a transparent yellow solution. The filtered solution is stored at 5 ° C ± 3 ° C for ~ 18 hours until further use.

[0363] The filtered bulk solution was thawed from 5°C ± 3°C to room temperature for 1 hour. The solution was a transparent dark yellow solution and did not contain any visible particles. The details of the diluent and IV (intravenous) kit are shown in Table 39 below. Table 40 shows the details of the dilution stability and IV kit compatibility of the injection formula (I) formate salt with different concentrations, and Table 41 shows the chemical analysis and particulate matter of the preparation.

[0364] Table 39 Diluent and IV kit details.

[0365]

[0366] Table 40: Dilution stability and IV kit compatibility of formate salts of Formula (I) for injection with different concentrations - Chemical analysis and particulate matter

[0367]

[0368]

[0369] Table 41: Results of In-Use Stability Study (Chemical Analysis)

[0370]

[0371] Table 42: Results of in-use stability study (impurity analysis)

[0372]

[0373]

[0374]

[0375] Table 43: Results of In-Use Stability Study (Particulate Matter)

[0376]

[0377] * Transparent slightly yellowish solution, no visible particles;

[0378] #Transparent colorless to slightly yellowish solution, no visible particles

[0379] @Placebo at the initial time point was on the higher side, probably due to vial contamination.

[0380] The formulations (bulk solutions) containing the formate salt of the compound of formula (I) for injection were diluted with 5% glucose injection at different concentrations. The diluted solutions were kept at room temperature (20°C-30°C) together with the IV kit. The different physicochemical parameters of these diluted solutions were evaluated, such as properties, pH, osmotic pressure, measured content of the formate salt of the compound of formula (I), related substances and particulate matter.

[0381] The dilution solution was found to be a transparent, slightly yellowish solution with no visible particles and remained stable for up to 8 hours at room temperature (20°C-30°C). The dilution solution showed a pH of 4.50 ± 0.20 and remained stable for up to 8 hours. As shown in Table 41, the dilution solution was essentially isotonic (270 to 330 mOsmol / kg) at a concentration of 1.25 mg / mL and remained stable for up to 24 hours. The remaining concentration dilution solutions (2.50, 3.50, 4.0 and 5.0 mg / mL) were slightly hypertonic solutions (370 to 460 mOsmol / kg), but this osmotic pressure range solution can be used as a large volume parenteral solution. The dilution solution remained stable for up to 8 hours, and no significant changes were observed in the measured content of the compound formate of formula (I). An increase in the level of impurities was observed from the initial (0 hour) to the 8 hour holding period, but the impurity level was within the specified limits of the bulk solution of the compound formate formulation of formula (I) for injection (total impurities: NMT 4%, Table 42). Particles in dilute solutions in accordance with USP <788> The limits mentioned for large volume parenterals in the chapter Particulate Matter in Injections. The initial (0 hour) particulate matter of the placebo (Table 43) did not meet the limits. This could be due to contamination that occurred during sample handling / sample analysis.

[0382] Based on in-use stability studies of formulations of the compound of formula (I) for injection (as bulk formate solution), it is recommended that bulk solutions of the compound of formula (I) for injection should be further diluted with 5% dextrose injection (freeflex bags) and used with the recommended IV kit within 6-8 hours after dilution when stored at room temperature (20°C-30°C) and protected from light. The following concentrations of the compound of formula (I) formate in dilution solutions using 5% dextrose injection are recommended for its administration: 1.25 mg / mL, 2.50 mg / mL, 3.50 mg / mL, 4.0 mg / mL, and 5.0 mg / mL.

[0383] Each placebo solution of different strengths should also be used with the recommended IV set within 6 hours of dilution when stored at room temperature (20°C-30°C) and protected from light.

[0384] Stability study of formulations

[0385] A pilot batch stability study of formate salt formulation of the compound of formula (I) for injection was prepared using the ingredients mentioned in Table 44 at 500 mg / vial and 1600 mL batch size.

[0386] Table 44: Formulation of formate salt of compound of formula (I) used for stability study SF20000011

[0387]

[0388]

[0389] Preparations are prepared by processes as described herein. Ascorbic acid is dissolved in water for injection (20% of the batch size, 25°C-30°C) and dissolved by stirring for 30 minutes, then lactic acid is added under stirring at room temperature for 15 minutes, and compound formate of formula (I) is added under stirring at room temperature for 150 minutes to obtain a first solution. Nicotinamide is added to the first solution and dissolved in 30 minutes at room temperature to obtain a second solution. Another 40% of WFI (of the total batch) is added to the second solution under stirring for 45 minutes, and L-arginine (pH regulator) is added under stirring at room temperature for 30 minutes to obtain a preparation. WFI is added to make up the volume of the preparation solution to the batch size. The solution is then filtered through a 0.2 μPES filter (47 mm, batch number 071715407, Sartorius) using a vacuum filtration assembly to obtain a preparation. 25.7 mL of the filtered solution is loaded into a 50 mL transparent USPI type tubular vial, the vial is half-stoppered and loaded for lyophilization. The lyophilization cycle used for the formulation was as follows.

[0390] Table 45: Lyophilization cycle formulation for formulations_Batch SF20000011

[0391]

[0392]

[0393] R: heating and cooling; H: maintaining

[0394] Total freeze drying cycle time: 112.83 hours

[0395] After lyophilization, the vacuum is broken with nitrogen, the vial is stoppered, unloaded from the lyophilizer, and sealed. The preparation is then dried to a pale yellow cake with a dark yellow bottom. The collapsed / melted cake dried at the corner of the vial. The lyophilized cake is easily reconstituted with water for injection within 3 minutes to form a transparent yellow solution. No pH change / pH movement of the reconstituted solution due to lyophilization is observed, and the results are shown in Table 46.

[0396] Table 146: Results of lyophilized cake of formulation - Batch No. SF20000011

[0397]

[0398]

[0399] The prepared batches were loaded for stability studies up to 12 months at 25°C ± 2°C / 60%RH ± 5%RH and 2°C - 8°C. Placebo batches (batch size: 400 mL) of each formulation were also manufactured and processed as per the active batches.

[0400]

[0401]

[0402] Sample 3M becomes a suspension. #Off-white to light yellow shrinkage at the bottom, yellow layer at the bottom of the vial; ##Light yellow to red cake at the bottom, with shrinkage; **Transparent yellow solution; ###Off-white to red cake, with shrinkage; Yellow layer at the bottom; ^Including 5 minutes of holding time after adding WFI; $Light yellowish to orange suspension; ~No 5 minutes of holding time; ￥Particle test: Diluted with 5D injection at a ratio of 1:5.

[0403] It can be inferred from Table 47 above that the color of the lyophilized cake was observed to change from yellow to reddish after 2 months at 25°C ± 2°C / 60% RH ± 5% RH and 2°C-8°C. No pH change of the formulation was observed under both stability conditions. A decrease in the assay content and an increase in impurities were observed at 25°C ± 2°C / 60% RH ± 5% RH. At 2°C-8°C, no significant differences were observed in the assay content and impurity profile. Based on the observation of the lyophilized cake of the formate salt formulation of the compound of formula (I) for injection at 500 mg / vial, further evaluation was performed at 250 mg / vial to improve the lyophilized cake structure to make it more aesthetically pleasing and pharmaceutically acceptable.

[0404] Formulation of formate salt of compound of formula (I) for injection for freeze-drying cycle validation and stability study 250 mg / vial

[0405] Table 15: Formulation of formate salt of compound of formula (I) for injection 250 mg / vial (batch size: 900 mL)

[0406]

[0407]

[0408] The preparations shown in Table 48 were prepared by the process described below. Ascorbic acid was dissolved in water for injection (20% of the batch size, 25°C-30°C) and dissolved by stirring for 30 minutes, then lactic acid was added under stirring at room temperature for 15 minutes, and compound formate of formula (I) was added under stirring at room temperature for 150 minutes to obtain a first solution. Nicotinamide was added to the first solution and dissolved by stirring for 30 minutes at room temperature to obtain a second solution. Another 40% (of the total batch size) of WFI was added to the second solution under stirring for 45 minutes, and L-arginine (pH regulator) was added under stirring at room temperature for 30 minutes. WFI was added to make up the volume of the solution to the batch size. The solution was then filtered through a 0.2 μPES filter (47 mm, batch number 071715407, Sartorius) using a vacuum filtration assembly to obtain a preparation. 25.7 mL of the filtered solution was loaded into a 50 mL transparent USPI type tubular vial, which was half-stoppered and loaded for lyophilization. The lyophilization cycles used for the formulations are provided in Table 49 below.

[0409] Table 16: Formulation of the lyophilization cycle of the formate salt of the compound of formula (I) (250 mg / vial)

[0410]

[0411]

[0412] R: temperature rise and temperature drop; H: maintain

[0413] Total freeze drying cycle time: 88.33 hours

[0414] The solubilization of the formate salt of the compound of formula (I) required a longer time, i.e., about 3 hours, and some dust-like particles were found to be visible. The unfiltered bulk solution was translucent yellow, but after filtering through a 0.2μ filter it was found to be a transparent yellow solution. The assay results of the unfiltered and filtered bulk solutions did not vary significantly.

[0415] After freeze drying, vacuum is broken with nitrogen, and the bottle is stoppered, unloaded and sealed from freeze dryer.Dry, complete, off-white to light yellow cake, cake bottom is dark yellow, and does not observe the contraction or remelting of freeze-dried cake.At the bottom of the bottle, yellow ring is observed, which may be caused by the phase separation of drug substance during freeze drying.11.5mL WFI is used to reconstruct the cake, and the resulting solution is a transparent yellow solution.In order to reconstruct, 11.5mL WFI is added to the bottle and then it is placed at room temperature for 5 minutes to ensure the proper wetting of solid, and then vigorously shaken 2-3 minutes.The complete cake on the top easily dissolves when adding WFI, without any shaking.The dry yellow layer at the bottom needs to be vigorously shaken to dissolve.

[0416] Table 50: Results of lyophilized cake of formate salt of compound of formula (I) at 250 mg / vial strength

[0417]

[0418] Extrapolation of the lyophilized formate salt of the compound of formula (I) at 250 mg / vial

[0419] A lyophilized formulation of the compound of formula (I) in a specification of 250 mg / vial, the lyophilization of the formulation results in the formation of a pharmaceutically acceptable lyophilized cake structure. The batch was sampled for a stability study of up to 24 months at 25°C ± 2°C / 60% RH ± 5% RH and 2°C-8°C. The placebo batch formulation was also manufactured and processed according to the active batch. A compatibility study of the manufacturing process and its process components was conducted, and the results obtained from all process components were satisfactory. Therefore, it can be concluded that the prepared formic acid salt formulation of the compound of formula (I) for injection is compatible with, for example, stainless steel 316L, USP type 1 glass, 0.2μ PES filter, 0.2μ PVDF filter, PharmaPure AL242029 tube and Sani Tech Tubes and other components are compatible for up to 24 hours.

[0420] It is to be understood that the reconstituted formulation of the formate salt of the compound of formula (I) for injection at 250 mg / vial should be used within 24 hours after its reconstitution when stored at room temperature.

[0421] The 250 mg / mL reconstituted solution of the formate salt of the compound of formula (I) for injection is further diluted with 5% dextrose injection IP and should be used within 10 hours after its dilution when stored at room temperature. Concentrations of the formate salt of the compound of formula (I) diluted with 5% dextrose injection IP, for example, 1.0 mg / mL and 3.33 mg / mL are recommended for its administration.

[0422] Temperature Excursion Studies and Freeze-Thaw Studies

[0423] Two different batches of freeze-dried formulations and placebo of the compound formate of formula (I) were manufactured for evaluating freeze-thaw and temperature excursion stability. For freeze-thaw cycle stability study, exposure conditions were -20°C for 24 hours, followed by 25°C ± 2°C / 60% ± 5%RH for 24 hours. The exposure was considered to have completed a freeze-thaw cycle. The preparation was exposed to a total of 3 freeze-thaw cycles together with secondary packaging. For freeze-dried formulations, that is, the compound formate of formula (I) for IV injection was exposed to respective storage conditions and respective duration together with secondary packaging. Exposure conditions were -20°C for 24 hours and 48 hours and 40°C ± 2°C / 75% ± 5%RH for 24 hours and 48 hours.

[0424] It was found that the formulation of the formate salt drug product of the compound of formula (I) for injection at 250 mg / vial was stable for up to 24 hours even when the formulation was exposed to different temperature conditions ranging from -20°C to 40°C ± 2°C.

[0425] It was also found that 250 mg / vial of the compound of formula (I) for injection formate salt was stable for up to 144 hours (3 cycles of freeze-thaw study duration) even when the formulation was exposed to different temperature conditions, i.e., -20°C ± 2°C to 25°C ± 2°C.

[0426] Photostability studies

[0427] Lyophilized formulations of the formate salt of the compound of formula (I) were manufactured as described herein. The formulations were packaged in various packages and exposed to light to assess their photostability.

[0428] The lyophilized formulation (250mg / vial) in 50mL transparent USP type 1 glass vial is kept as is for light exposure until the required total illumination is reached. In another embodiment, the lyophilized formulation in 50mL transparent USP type 1 glass vial is kept in a secondary carton and exposed to a light source until the required total illumination is reached. Similarly, the preparation is packaged in aluminum foil and a carton and subjected to illumination. For comparison purposes, control samples and placebo are also subjected to illumination. Table 51A shows the details of the packaging materials used. Similarly, only the active component, i.e., the compound formate of formula (I), is subjected to light stability studies and the detailed information of the packaging materials is shown in Table 51B.

[0429] Table 51A: Details of packaging materials used:

[0430]

[0431] Table 51B: Details of packaging materials used

[0432]

[0433] The photostability of formulations for injection comprising the formate salt of the compound of formula (I) and the formate salt of the compound of formula (I) alone was evaluated according to the ICH-Q1B guidelines.

[0434] The formulation of the formate salt of the compound of formula (I) for injection at 250 mg / vial was found to be insensitive to light. The remaining components in the formulation provided a photoprotective effect by forming a solid cake structure at the bottom of the lyophilized cake, above the API layer.

[0435] Photostability studies have shown that the compound of formula (I) drug as a formate salt was found to be light sensitive, as discoloration was observed. Therefore, the compound of formula (I) formate salt should be protected from light during the manufacturing process of the formulation. However, the formulation was found to be light insensitive. Therefore, the formulation of the present disclosure is light stable and enhances the stability of the compound of formula (I) formate salt.

[0436] Vmax Research

[0437] Volume Maximization Study (Vmax) The purpose of the constant pressure study is to determine the sterilizing grade filter and membrane-based pre-filter capacity and to provide dimensions for the sterilizing grade filter and membrane-based pre-filter for a given unit operation.

[0438] Virus Vmax container is used to test under manual operation, wherein filter is connected to Virus Vmax container with the help of Luer lock (luer-lock) connection.Before operation, the filter is moistened / adjusted with water for injection and buffer.In addition, before this process, the device is properly ventilated to avoid any air entrapment.The cumulative volume measurement value as a function of time is recorded during the experiment, which is used as a measure (under constant pressure) of filter performance in turn.T / V (time / volume) as a function of time (t) is plotted to plot slope, to calculate the capacity (Vmax) and initial volume flow (Ji) of the filter based on the gradual pore plugging model (gradual poreplugging model).The minimum filtration area (A minute) required for the batch volume (VB) in time (tB) is calculated using the following formula.

[0439]

[0440] In the above equation, V = VB = process volume (L)

[0441] Filters evaluated: Sartoguard 1.2μ / 0.2μ PES nominal filter as pre-filter; followed by Sartopore 0.2μ PES absolute filter as sterile filter as shown in Table 51C.

[0442] Table 51C

[0443]

[0444] The formulation of the compound formate salt of formula (I) for injection at 250 mg / vial was found to be easily filtered. No filter clogging and flow attenuation were observed. The results (Table 51D) show that the formulation of the compound formate salt drug product of formula (I) was not adsorbed on the filter. There was no significant change in related substances.

[0445] Table 51D: V max Results of the study

[0446]

[0447] Observations on the formulations of the present disclosure

[0448] As can be understood from the above examples on developing formulations of the compound formate of formula (I), the drug product, i.e. the compound formate of formula (I), is unstable in solution for a long time, and therefore it is necessary to prepare it as a formulation and lyophilize the formulation to improve stability.

[0449] Further, based on the 6-month stability data of the prototype formulation batch, it was found that the formulation of the compound formate of formula (I) for injection was stable at 2°C-8°C. As the pH was reduced from pH 4.50 to pH 3.50, the impurity spectrum of the formulation was also reduced at higher temperatures, i.e., at 40°C ± 2°C / 75% RH ± 5% RH and at 25°C ± 2°C / 60% RH ± 5% RH. In addition, formulations with l-arginine in the specifications of 500 mg / vial and 250 mg / vial were used for long-term stability studies. The formulation also contains a combination of two acids, i.e., ascorbic acid and lactic acid. Further, in order to reduce the acid concentration in the formulation, l-arginine was added. The specification was reduced from 500 mg / vial to 250 mg / vial to improve the aesthetics of the freeze-dried cake structure with pharmaceutical acceptability and produce freeze-dried cakes using a freeze-drying cycle time period of 3-4 days as disclosed herein.

[0450] In order to use the preparation in injection, the lyophilized preparation is reconstituted with a reconstitution agent and a diluent. The reconstitution agent is water for injection, and, for dilution, 5% dextrose (diluent) is used. It is found that the reconstituted preparation is stable and compatible for up to 10 hours after dilution at room temperature. When dilution is required, 1.0 mg / mL and 3.33 mg / mL dilution preparations can be used. For the preparation of the compound formate drug product containing 20 mg / mL formula (I), the product specification target is 250 mg / vial. Therefore, the preparation of the present disclosure is provided in the following table 52.

[0451] Table 52: Formulation of the formate salt drug product for injection 250 mg / vial

[0452]

[0453] A kit comprising the formulation.

[0454] The preparation of the present disclosure can be used in the form of a kit, wherein the kit comprises a first compartment and a second compartment. The first compartment comprises the prepared preparation. In another embodiment, the kit comprises a reconstituted preparation or a lyophilized preparation. The second compartment comprises water. The kit optionally comprises an accessory selected from a diluent bag, an infusion tube, a needle or a connector.

[0455] Treatments for bacterial infections

[0456] Dose-finding study of the formulation in a neutropenic murine model of thigh infection caused by A. baumannii ATCC 17978

[0457] The purpose of this study was to investigate the efficacy of formulations comprising the formate salt of the compound of formula (I) as disclosed herein administered by subcutaneous (SC) injection every 8 hours (q8h) at a dose range of 10 mg / kg to 120 mg / kg using a 26-hour neutropenic mouse bilateral thigh infection model infected with Acinetobacter baumannii ATCC 17978. Established antibiotics meropenem and polymyxin B were used as comparative controls.

[0458] Two hours before treatment with the compound formate of formula (I) administered subcutaneously (SC) at 10, 30, 60, 90 and 120 mg / kg q8h and comparative controls meropenem (administered SC at 200 mg / kg q8h) and polymyxin B (administered SC at 25 mg / kg q8h), mice were infected intramuscularly with Acinetobacter baumannii ATCC 17978. Animals were euthanized at 2 hours post-infection (pre-treatment control) or 26 hours post-infection (vehicle control and all treatment groups). Data were obtained from all animals at early time points.

[0459] The lyophilized vials of the formate salt of the compound of formula (I) as formulation were reconstituted with SWFI and the reconstituted solution was diluted with SWFI to the desired dosage strength for administration via the subcutaneous route. Similarly, for the dose control group animals, the lyophilized placebo vials were reconstituted with SWFI and diluted with SWFI to the desired strength.

[0460] In the neutropenic rat thigh model, when evaluating the SC dose range of 10 to 120 mg / kg q8h, the formate salt of the compound of formula (I) effectively reduced the bacterial burden of Acinetobacter baumannii ATCC 17978 at 10 mg / kg and 30 mg / kg q8h, showed an inhibitory or bactericidal effect on Acinetobacter baumannii ATCC 17978 at a dose of 60 mg / kg q8h, and achieved logarithmic killing at higher doses of 90 mg / kg and 120 mg / kg q8h.

[0461] The formate salt of the compound of formula (I) at doses ≥ 90 mg / kg q8h was more effective against this strain of Acinetobacter baumannii than polymyxin B (administered SC at 25 mg / kg q8h) and subcutaneous meropenem (200 mg / kg q8h). Figure 4 The CFU / g of rat thighs infected with Acinetobacter baumannii ATCC 17978 and treated subcutaneously with [compound formate of formula (I)] or meropenem (200 mg / kg) or polymyxin B (25 mg / kg) q8h (every 8 hours) are depicted. The neutropenic rat double thigh model of Acinetobacter baumannii ATCC17978 was used to demonstrate the efficacy of the compound formate of formula (I) when administered subcutaneously in vivo. When the SC dose range of 10 to 120 mg / kg q8h was evaluated in three replicate studies, the compound formate of formula (I) effectively reduced the bacterial load of Acinetobacter baumannii ATCC 17978 at 10 mg / kg and 30 mg / kg q8h and caused logarithmic killing in a general dose-dependent manner at ≥60 mg / kg q8h.

[0462] Thus, in the neutropenic rat thigh model, a formulation comprising the formate salt of the compound of formula (I) exhibited an inhibitory or killing effect on Acinetobacter baumannii ATCC 17978 at a dose of 60 mg / kg q8h, achieving logarithmic killing at higher doses of 90 mg / kg and 120 mg / kg q8h. The formulation of the present disclosure was more effective against this Acinetobacter baumannii strain at a dose of ≥90 mg / kg q8h than polymyxin B (administered SC at 25 mg / kg q8h) and subcutaneous meropenem (200 mg / kg q8h).

[0463] Advantages of the present disclosure

[0464] The present disclosure provides a formulation comprising a compound formate of formula (I) with a solubilizing agent, a pH adjusting agent and a hydrotrope. The formulation of the present disclosure is stable over a long period of time and is stable in a pH range of 2 to 6. The formulation of the present disclosure is stable at temperatures up to -25°C to 10°C. In addition, the formulation of the present disclosure can be lyophilized and stability can be improved. The present disclosure also provides a reconstituted formulation that makes the formulation in an injectable form. The present disclosure further provides a process for preparing a formulation that is easy to adapt and suitable for large-scale production. The present disclosure also provides a formulation capable of treating infections caused by microorganisms. The present disclosure also provides a method for treating or preventing a disease or condition mediated by a microorganism and the disease or condition is selected from complicated and uncomplicated urinary tract infections (cUTI, such as pyelonephritis, cystitis), intra-abdominal infections (cIAI), bloodstream infections, hospital-acquired bacterial pneumonia (HABP, nosocomial pneumonia), ventilator-associated bacterial pneumonia (VABP), community-acquired bacterial pneumonia (CABP), cystic fibrosis secondary infection (CFI), skin and soft tissue infections (SSTI), endocarditis, brain meningitis, dysentery and diarrhea, typhoid fever, Clostridium difficile-associated colitis and diarrhea (CDAD), Helicobacter pylori-associated peptic ulcer, sexually transmitted diseases caused by Chlamydia trachomatis, gonorrhea caused by Neisseria gonorrhoeae, syphilis caused by Treponema pallidum, or bioterrorism-related bacterial diseases caused by anthrax (Bacillus anthracis), plague (Yersinia pestis), tularemia (Franciscoides tularensis), glanders (Burkholderia mallei), melioidosis (Burkholderia pseudomallei), and Q fever (Coxiella burnetii).

Claims

1. A preparation comprising: a. a compound of formula (I) or a salt thereof; b. solubilizing agent; c. hydrotropes; and d. pH adjuster.

2. A formulation as claimed in claim 1, wherein the formulation comprises a compound of formula (I) or a salt thereof in an amount ranging from 1 to 30% (w / w) by weight; a solubilising agent in an amount ranging from 5 to 55% (w / w) by weight; a hydrotrope in an amount ranging from 1 to 30% (w / w) by weight; and a pH adjusting agent in an amount ranging from 0.5 to 30% (w / w) by weight.

3. The formulation of claim 1, wherein the solubilizing agent is selected from lactic acid, ascorbic acid, acetic acid, propionic acid, succinic acid, gluconic acid, benzoic acid, tartaric acid, glutaric acid, malic acid, fumaric acid or a combination thereof; the pH adjusting agent is selected from potassium hydroxide, sodium hydroxide, l-arginine, histidine, glycine, sodium bicarbonate or a combination thereof; the hydrotrope is selected from nicotinamide, sodium benzoate, sodium citrate, sodium acetate or a combination thereof; and the salt of the compound of formula (I) is selected from formate, methanesulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, acetate, propionate, fumarate, maleate, tartrate, succinate, glycolate, glutamate, aspartate, hydrochloride, hydrobromide or sulfate.

4. The formulation of claim 1, wherein the formulation is stable at a temperature ranging from -70°C to 10°C.

5. The formulation of claim 1, wherein the pH of the formulation is in the range of 2 to 6.

6. The formulation of claim 1, wherein the formulation comprises a vehicle; and the vehicle is water.

7. The formulation of claim 1, wherein the formulation is capable of killing or inhibiting the growth of microorganisms; and the microorganisms are selected from bacteria, viruses, fungi, and protozoa.

8. The preparation of claim 1, wherein the preparation is an anti-infective agent against microorganisms.

9. The preparation of any one of claims 7 to 8, wherein the microorganism is selected from Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Enterobacter cloacae, Citrobacter, Proteus, Serratia marcescens, Salmonella, Morganella, Klebsiella oxytoca, Klebsiella aerogenes, Providencia, Neisseria gonorrhoeae, Mycoplasma, Campylobacter, Fusobacterium, Bacteroides fragilis, Prevotella, Shigella, Helicobacter pylori pylori, Ureaplasma spp., Burkholderia gladiolus, Burkholderia multivora, Pandoraella, Burkholderia cepacia, Burkholderia pseudomallei, Burkholderia mallei, Stenotrophomonas maltophilia, Achromobacter spp., Ralstonia pietermaritzia, Legionella pneumophila, Clostridium difficile, Staphylococcus aureus, coagulase-negative staphylococci, Streptococcus pyogenes, Streptococcus pneumoniae, Enterococcus faecium, Enterococcus faecalis, Bacillus anthracis, Yersinia pestis, and Francisella tularensis.

10. The formulation of claim 1, wherein the formulation inhibits an enzyme selected from bacterial gyrase, topoisomerase IV, or a combination thereof.

11. The formulation of claim 1, wherein the formulation further comprises an additive, a therapeutic agent, or a combination thereof.

12. The formulation of claim 11, wherein the additive is selected from sorbitol, dextrose, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium citrate, sodium bicarbonate, sodium chloride, or a combination thereof.

13. A lyophilized preparation, comprising: the preparation according to claim 1.

14. A reconstituted formulation comprising: the lyophilized formulation according to claim 13 or the formulation according to claim 1, and a reconstitution agent and a diluent.

15. The reconstituted formulation of claim 14, wherein the reconstitution agent is water; the diluent is selected from dextrose, sorbitol, sodium bicarbonate, glucose, mannitol, sucrose or sodium chloride; and the weight range of the diluent in the first solvent is 0.9-25% (w / v).

16. The formulation of claim 15, wherein the first solvent is water.

17. A kit comprising: a. A first compartment comprising a formulation as claimed in claim 1 or 13 or 14; b. a second compartment comprising a facilitator; and c. Optional accessories.

18. The kit of claim 17, wherein the accelerator is selected from water for injection; and the accessories are selected from diluent bags, infusion tubes, needles and connectors.

19. A process for preparing the formulation of claim 1, comprising: a. contacting a solubilizing agent with a compound of formula (I) or a salt thereof in the presence of a second solvent under stirring at a temperature in the range of 18 to 40 ° C to obtain a first solution; b. adding a hydrotrope to the first solution under stirring to obtain a second solution; and c. Mixing a pH adjuster into the second solution, and then optionally adding a vehicle, to obtain the formulation.

20. A process as claimed in claim 19, wherein two or more solubilising agents are mixed prior to contact with the compound of formula (I) or a salt thereof.

21. The process of claim 19, wherein the pH of the first solution is in the range of 2 to 3; and the pH of the second solution is in the range of 2.5 to 4.

22. The process of claim 19, wherein the preparation is filtered after adding the solvent.

23. The process of claim 19, wherein the second solvent is water; and the vehicle is water.

24. A process for preparing the lyophilized formulation of claim 19, comprising: a) freezing the preparation according to claim 1 at a temperature ranging from 0 to -50°C, and then drying at a pressure ranging from 10 μbar to 1 bar to obtain the lyophilized preparation.

25. The process of claim 24, wherein freezing is performed for a period of time ranging from 30 minutes to 40 hours; and drying is performed for a period of time ranging from 20 minutes to 150 hours.

26. A process for preparing the reconstituted formulation of claim 14, comprising: The lyophilized formulation of claim 13 is contacted with a diluent and then a reconstitution agent is added.

27. The process of claim 26, wherein the process is performed for a period of time in the range of 60 seconds to 10 minutes.

28. The process of claim 26, wherein the reconstituted formulation is stable at a temperature in the range of 20 to 35°C for a period of time in the range of 20 to 30 hours.

29. The process of claim 26, wherein the pH of the reconstituted formulation is in the range of 3 to 5 and the osmotic pressure is in the range of 300-500 mOs-mol / kg.

30. Use of a formulation as claimed in claim 1 or claim 13 or claim 14.

31. Use of the kit according to claim 17.

32. A method of treating a microbial infection, the method comprising administering to a subject in need thereof an effective amount of the formulation of claim 1 or claim 13 or claim 14.

33. The method of claim 32, wherein the microbial infection is caused by bacteria, viruses, fungi, or protozoa.

34. A method of treating or preventing a disease or condition, the method comprising administering to a subject in need thereof an effective amount of the formulation of claim 1 or claim 13 or claim 14.

35. The method of claim 34, wherein the disease or condition is mediated by a Gram-positive bacterial species, a Gram-negative bacterial species, or a combination thereof.

36. The method of claim 34, wherein the disease or disorder is selected from complicated and uncomplicated urinary tract infections (cUTI, e.g., pyelonephritis, cystitis), intra-abdominal infections (cIAI), bloodstream infections, hospital-acquired bacterial pneumonia (HABP, nosocomial pneumonia), ventilator-associated bacterial pneumonia (VABP), community-acquired bacterial pneumonia (CABP), secondary infections of cystic fibrosis (CFI), skin and soft tissue infections (SSTI), endocarditis, meningitis, dysentery and diarrhea, typhoid fever, Clostridium difficile-associated colitis and diarrhea (CDAD), Helicobacter pylori-associated peptic ulcers, sexually transmitted diseases caused by Chlamydia trachomatis, gonorrhea caused by Neisseria gonorrhoeae, syphilis caused by Treponema pallidum, or bioterrorism-related bacterial diseases caused by anthrax (Bacillus anthracis), plague (Yersinia pestis), tularemia (Franciscoides tularensis), glanders (Burkholderia mallei), melioidosis (Burkholderia pseudomallei), or Q fever (Coxiella burnetii).

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