Solid dispersions comprising amorphous 2-[3-[4-(1h-indazol-5-ylamino) quinazolin-2-yl] phenoxy]-n-propyl-2-yl-acetamide

Abstract

The present application relates to a solid dispersion comprising amorphous besudil. Also provided are pharmaceutical dosage forms comprising the solid dispersion comprising amorphous besudil as an active pharmaceutical ingredient, and methods of using the pharmaceutical dosage forms in the treatment of diseases or disorders modulated by ROCK.

Description

Technical Field

[0001] This application relates to solid dispersions comprising amorphous 2-{3-[4-(1H-indazol-5-ylamino)-2-quinazolinyl]phenoxy}-N-(propan-2-yl)acetamide (also known as belumosudil and KD025). This application further relates to methods of preparing solid dispersions comprising amorphous belumosudil as described herein, pharmaceutical compositions comprising one or more of said solid dispersions, and methods of using said pharmaceutical compositions to treat diseases and conditions regulated by Rho-associated coiled-coil forming protein kinase (ROCK). Background Art

[0002] Belumosudil, chemically known as 2-{3-[4-(1H-indazol-5-ylamino)-2-quinazolinyl]phenoxy}-N-(propan-2-yl)acetamide, is represented by Formula I:

[0003] Belumosudil (also known as KD025) is an inhibitor of Rho-associated coiled-coil forming protein kinase (ROCK). Belumosudil binds to and inhibits the serine / threonine kinase activities of ROCK1 and ROCK2, and thus can be used to treat diseases, disorders and conditions regulated by ROCK, particularly including autoimmune or fibrotic diseases, acute and chronic graft-versus-host disease (GVHD), idiopathic pulmonary fibrosis, and moderate to severe psoriasis and other indications. The mesylate salt of belumosudil is currently sold in the United States and other countries under the trade name (Kadmon Corp. / Sanofi) for the treatment of patients with chronic GVHD (cGVHD), in some cases for the treatment of patients with chronic GVHD (cGVHD) after failure of at least two prior lines of systemic therapy.

[0004] A method for preparing belumosudil is disclosed in U.S. Patent 8,357,693 ('693 patent), specifically in Example 82 thereof. The method disclosed in the '693 patent provides belumosudil as a crude product, which is purified by high performance liquid chromatography (HPLC). Belumosudil and methods for preparing said compound are also described in U.S. Patent No. 9,815,820, U.S. Patent No. 10,183,931, and U.S. Patent No. 10,696,660.

[0005] Bezudil is a weakly basic compound that is poorly soluble in water. Current methods of administering bezudil include formulating the mesylate salt of bezudil into pharmaceutically acceptable capsules and tablets for oral administration. Given the low water solubility of bezudil, the compound tends to precipitate when transitioning from the acidic gastric environment of the stomach and early digestive pathways to the more neutral pH of the intestinal environment. Thus, the low solubility of bezudil may affect the manner and timing of its bioabsorption as well as its pharmacokinetic properties. In addition, the low solubility of bezudil may limit the use of traditional excipients and wet granulation methods.

[0006] The low solubility and variable pharmacokinetic properties of bezudil present challenges in the development of alternative formulations. For illustration, a formulation of bezudil with enhanced solubility would provide greater flexibility and a wider range of options in developing different dosing regimens, formulations, and modes for delivering the compound to a subject.

[0007] Accordingly, there remains a need for formulations comprising bezudil that can address these challenges. SUMMARY OF THE INVENTION

[0008] In one aspect, the present application provides a solid dispersion comprising bezudil in an amorphous form. In some embodiments, the amorphous form of bezudil is placed in a solid dispersion comprising a matrix carrier material. The use of amorphous bezudil in a solid dispersion enhances its solubility and improves its biological properties, thereby providing an extended method for formulating and delivering a drug.

[0009] In some embodiments, the present application provides an amorphous solid dispersion comprising bezudil, optionally prepared using spray drying techniques.

[0010] In some embodiments, the amorphous form of bezudil is provided as an amorphous solid dispersion of bezudil formulated with at least one polymer (optionally, e.g., a pharmaceutically acceptable polymer).

[0011] Another aspect of the present application provides a method for preparing a solid dispersion comprising amorphous bezudil. Solvent evaporation methods (such as spray drying) can be used to prepare a solid dispersion comprising amorphous bezudil. A solid dispersion comprising amorphous bezudil can be prepared by the following process: dissolving bezudil in a suitable solvent; adding one or more carrier matrix materials; and removing the solvent by spray drying, thereby providing a solid dispersion of amorphous bezudil contained in the carrier matrix material.

[0012] Solid dispersions containing amorphous besudil can be used to prepare solid pharmaceutical dosage forms, such as tablets and capsules. A pharmaceutical composition containing an effective amount of amorphous besudil can be used to treat ROCK-regulated diseases, disorders, and conditions as further described in this application. Description of the Drawings

[0013] Figure 1 Scanning electron microscope (SEM) images of the solid dispersions of Example 1 (specifically Examples 1.1, 1.2, and 1.3 in Table 5) captured at magnifications of 1500x and 5000x are shown.

[0014] Figure 2 XRPD results of six spray-dried formulations prepared as in Example 1 are shown.

[0015] Figure 3 Two-stage dissolution data of six spray-dried besudil formulations of Example 1 are shown compared to crystalline besudil mesylate form.

[0016] Figure 4 is Figure 3 An expanded view of a portion of the data.

[0017] Figure 5 XRPD results of three solid dispersions of Example 3 are shown.

[0018] Figure 6 SEM images of three solid dispersions (Formulations F1, F2, and F3) of Example 3 captured at magnifications of 1500x and 5000x are shown.

[0019] Figure 7 Particle size data of three solid dispersions (Formulations F1, F2, and F3) of Example 3 are shown.

[0020] Figure 8 Modulated differential scanning calorimetry (mDSC) evaluation of Tg for three solid dispersions (Formulations F1, F2, and F3) of Example 3 after one cooling cycle (upper profile) and one heating cycle (lower profile) with a homogeneous determination is shown.

[0021] Figure 9 Non-sink dissolution data of three solid dispersions (Formulations F1, F2, and F3) of Example 3 are shown compared to crystalline besudil mesylate form.

[0022] Figure 10Shows the assay and impurity data of the three solid dispersions (Formulations F1, F2, and F3) of Example 3 compared to the crystalline mesylate form of besulifloxacin and the diluent described in Example 4.

[0023] Figures 11A - 11C Shows the XRPD diffractograms of the three solid dispersions of Example 3 after an 8-week stability study as described in Example 5 ( Figure 11A : F1 [20:80 KD025:PPPEG]; Figure 11B : F2 [20:80 KD025:PVPVA]; and Figure 11C : F3 [40:60 KD025:PPPEG]).

[0024] Figure 12 Shows polarized light microscopy images (5X magnification) of the solid dispersions (F1, F2, and F3) of Example 3 at 25 mg A / mL in a 0.5 wt.% Methocel A4M (aqueous) suspension.

[0025] Figure 13 Shows a graph of the in vivo plasma concentration (ng / mL) of besulifloxacin versus time after administration of besulifloxacin as described in Example 6: (a) tablet formulation - fasting; (b) F2 (20:80 KD025:PVPVA) - fasting; (c) tablet formulation - fed; and (d) F2 (20:80 KD025:PVPVA) - fed. Detailed Description

[0026] The present application provides solid dispersions comprising an amorphous form of besulifloxacin, which improve the solubility of the compound and its biological properties. Reliable dosing and absorption of besulifloxacin are important for ensuring consistent systemic exposure. Therefore, the development of a reproducible drug delivery system and the characterization of its associated dissolution profiles provide an advantage in ensuring consistent and effective dosing of besulifloxacin.

[0027] Solvent evaporation methods such as spray drying can be used to prepare solid dispersions comprising amorphous besulifloxacin. The technique involves dissolving or suspending besulifloxacin in a polymeric matrix carrier, followed by spraying the mixture stream. Spray drying removes the solvent, resulting in a solid dispersion comprising amorphous besulifloxacin dispersed in a polymeric matrix carrier.

[0028] Using spray drying technology, besudil is converted into an amorphous state dispersed in a polymer matrix carrier. The amorphous besudil prepared by the method disclosed in this application provides dissolution enhancement by reducing particle size and removing the crystal lattice. The lack of crystallinity enables besudil to dissolve without overcoming lattice energy. The carrier material can additionally assist dissolution by improving the wetting, solubility, and stability characteristics of besudil in supersaturated solutions. The spray-dried solid dispersion containing amorphous besudil provides good physicochemical properties (such as controlled particle size and flowability), which can be used for downstream processing, such as tableting.

[0029] In addition, compared to a composition containing crystalline besudil, a pharmaceutical composition containing a solid dispersion containing amorphous besudil demonstrated increased dissolution; in some embodiments, a significant increase in dissolution was obtained with the solid dispersion of this application. The improved solubility of the solid dispersion containing amorphous besudil provides advantages and greater flexibility for formulation development and drug delivery. Definitions

[0030] As used in this application, the term besudil (or KD025) refers to 2-{3-[4-(1H-indazol-5-ylamino)-2-quinazolinyl]phenoxy}-N-(propan-2-yl)acetamide represented by the following formula I:

[0031] In some embodiments, amorphous besudil is in the free base form.

[0032] When the term "besudil" is used in this application, it should be understood that unless the context clearly indicates otherwise, the term can cover any form of the compound besudil and its pharmaceutically acceptable salts. The term "besudil" refers to both the compound besudil (e.g., free base form, amorphous form, or crystalline form), the pharmaceutically acceptable salts of besudil (e.g., the mesylate form as used in REZUROCK TM ), and any form of besudil that can be used in a formulation or pharmaceutical composition for administering the compound to a patient.

[0033] The term "pharmaceutically acceptable salts" refers to non-toxic inorganic and organic acid addition salts of besudil. In some embodiments, the pharmaceutically acceptable salt of besudil in this application is mesylate.

[0034] As used in this application, "about" includes the exact amount modified by the term about and amounts expected to be within experimental error (e.g., within 15%, 10%, or 5%). For example, "about 5 mg" means "5 mg" and the range of mg within experimental error (e.g., 5 mg ± 15%, 10%, or 5%). As used in this application, the term "about" can be used to modify ranges as well as specific values.

[0035] The acronym API refers to "active pharmaceutical ingredient", which is synonymous with the definition of besudil (or KD025) and its pharmaceutically acceptable salts (optionally, the mesylate salt of besudil) when used in this application.

[0036] As used in this application, "administering" or "administer to" refers to the act of prescribing one or more drugs containing the API for a subject to take during treatment, the act of dispensing the one or more drugs to the subject, and / or the act of physically receiving or ingesting the one or more drugs. Thus, the API (besudil) can be "administered" by a physician or other medical personnel who writes a prescription for one or more drugs; and / or by a pharmacist who fills the prescription and / or dispenses the one or more drugs to the subject; and / or by a patient or subject who ingests the drug and / or his or her partner or caregiver who provides the drug to the subject.

[0037] As used in this application, the term "solid dispersion" refers to a system in which the API is dispersed throughout a solid carrier (in some embodiments, a solid matrix carrier). In some embodiments, the carrier comprises small molecules and / or polymers or copolymers, optionally polymers. Thus, the solid dispersion will include at least two components, one of which is the API and the other is the carrier. Optionally, additional additives (such as surfactants) may be included. Optionally, in the solid dispersion, the API is homogeneously or uniformly dispersed throughout the carrier matrix.

[0038] As used in this application, the term "amorphous solid dispersion" refers to a single-phase amorphous system in which the API is dispersed or dissolved in the carrier matrix (optionally, a polymer matrix) in a molecular state.

[0039] In some embodiments of this application, the ratio of besudil to one or more carrier matrix materials in the solid dispersion can be about 10:90 to 90:10 by weight; or about 20:80 to 80:20 by weight; or about 25:75 to 75:25 by weight; or 40:60 to 60:40 by weight.

[0040] Unless otherwise specified, the term "amorphous" or "amorphous form" means that a substance or component is substantially non-crystalline and is in a disordered solid form, i.e., a solid form that is substantially lacking in long-range lattice order as determined by XRPD data. The substantially amorphous state includes at least about 50 wt%, optionally at least about 60 wt%, optionally at least about 70 wt%, optionally at least about 80 wt%, optionally at least about 90 wt%, optionally at least about 95 wt%, or optionally at least 99 wt% amorphous form of the API compared to other forms of the substance or component.

[0041] In some embodiments, amorphous besulforhodamine is a solid form of substantially amorphous besulforhodamine; in some embodiments, it is in a form comprising at least about 95% amorphous form of besulforhodamine; in some embodiments, it is in a form comprising at least 98% amorphous form of besulforhodamine. Whether besulforhodamine is in an amorphous form can be characterized, for example, by XRPD techniques as described in this application or by additional techniques known to those skilled in the art.

[0042] As used in this application, "carrier matrix" or "carrier matrix material" refers to a component that stabilizes, suspends, and / or transports amorphous form besulforhodamine when in the solid state. Optionally, the carrier matrix material is an amorphous polymer material. The choice of polymer in a solid dispersion can play an important role in the overall final product properties. Polymers used as carrier matrix materials can include polyvinylpyrrolidone derivatives such as polyvinylpyrrolidone (PVP) and polyvinylpyrrolidone-vinyl acetate copolymer (PVPVA) (such as those sold under the trade name Kollidon VA those sold); polymethacrylate derivatives (such as series); polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (PPPEG) (such as those currently sold under the trade name those sold); hydroxypropylmethylcellulose (HPMC) and hypromellose acetate succinate (HPMCAS). Optional carrier matrix materials for this application are PVPVA and PPPEG.

[0043] The term "effective amount" when used in connection with the amorphous form of besulforhodamine means an amount capable of treating or preventing a disorder, disease or condition or its symptoms as disclosed in the present application. For example, in a pharmaceutical composition, the effective amount of the amorphous form of besulforhodamine may be at a level that will provide the desired effect; for example, in a unit dose for oral administration, about 0.5 to 15 mg / kg of subject body weight, optionally about 1 to 5 mg / kg of subject body weight, optionally about 3 mg / kg of patient body weight. For example, the dose of besulforhodamine may be the currently therapeutic dose of 200 mg administered daily, or alternatively, a dose in the range of 10 mg to up to 1000 mg, optionally, for an adult patient, in the range of 100 mg to 400 mg; optionally, in the range of 100 mg to 200 mg.

[0044] In addition, optionally for a pediatric patient, the dose of besulforhodamine may be in the range of 10 mg to 200 mg. The dose of besulforhodamine can be adjusted according to the patient's body weight. For example, for a pediatric patient with a body weight in the range of about 6 kg to less than 20 kg, the dose may be in the range of about 10 to 50 mg, administered once daily; in another embodiment, for a pediatric patient with a body weight in the range of about 10 kg to less than 20 kg, the dose may be about 50 mg, administered once daily; for a pediatric patient with a body weight in the range of about 20 kg to less than 40 kg, the dose may be about 100 mg, once daily; for a pediatric patient with a body weight equal to or greater than 40 kg, the dose may be 200 mg, once daily.

[0045] It will be apparent to those skilled in the art that the effective amount of the amorphous form of besulforhodamine disclosed in the present application is expected to vary depending on: the severity of the indication being treated, the route of administration, and / or other drugs administered to the subject taking into account drug interactions (e.g., proton pump inhibitors or CYP3A inducers).

[0046] As used in the present application, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or vehicle, such as a solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc, magnesium stearate, calcium stearate or zinc stearate, or stearic acid). Each carrier must be "acceptable" in terms of being compatible with the other ingredients of the formulation (e.g., including the API so as not to slowly crystallize over time) and maintaining the stability of the other ingredients, and be harmless to the patient.

[0047] "Suitable solvent" as used in this application, such as the "suitable solvent" for dissolving besuldiol in a solvent system containing a carrier matrix material, means a solvent or a mixture of one or more solvents that is compatible with besuldiol and one or more carrier matrix materials and is capable of sufficiently dissolving besuldiol and said one or more carrier matrix materials to enable the use of spray drying techniques. The term "suitable solvent" may include a mixture of solvents and thus may be interchangeable with "suitable solvent system". The solubility of the API and / or carrier matrix material can be confirmed by filtration and HPLC analysis or by visual observation (e.g., a clear or substantially clear solution is produced upon visual observation). "Suitable" for a solvent also means that the one or more solvents do not present unacceptable toxicity or environmental hazards and are acceptable when used in the manufacture of a drug for human consumption.

[0048] Unless the context otherwise requires, "or" is used in an inclusive sense (equivalent to "and / or"). General methods of preparation and use

[0049] A solid dispersion containing amorphous besuldiol can be prepared by the following process: dissolving besuldiol in a suitable solvent; adding one or more carrier matrix materials to the besuldiol solution; and removing the solvent, or substantially removing the solvent, to provide amorphous besuldiol dispersed in the carrier matrix.

[0050] In the first step of the method involving adding besuldiol to a suitable solvent, besuldiol can be in various forms, e.g., any polymorphic crystalline form or solvate. Besuldiol can be in salt form or can be in free base form. If besuldiol is in the form of an acid addition salt and amorphous besuldiol in free base form is desired, then a sufficient amount of base can be added to the solvent to form besuldiol free base. The base can be an inorganic base such as an alkali metal hydroxide, or the base can be an amine such as diethylamine, triethylamine, etc.

[0051] In the context of this method, the choice of solvent is an important consideration in the preparation of amorphous solid dispersions, and combinations of solvents can optionally be used to obtain the desired solvent parameters. As described in Example 1 of the present application, a large number of solvent screening experiments were conducted to obtain a solvent system that can be used to dissolve besudil and the carrier matrix material, thereby enabling the use of spray drying technology. After these solubility experiments, it was found that a suitable solvent system for preparing amorphous besudil via spray drying comprises a mixture of triethylamine (TEA) and acetone. In contrast, solvent systems comprising the following do not effectively dissolve besudil and / or besudil was determined to be sparingly soluble or slightly soluble in one or more of these solvents, such that these solvents and / or solvent systems are not "suitable solvents" as defined in the present application, the solvent systems comprising: acetone, ethyl acetate, acetonitrile (ACN), tetrahydrofuran, methanol, dichloromethane (DCM), dimethylformamide (DMF), isopropanol (IPA), methyl ethyl ketone, methyl isobutyl ketone (MIBK), methyl tert-butyl ether (MTBD), n-heptane, toluene, a mixture of DCM and methanol, a mixture of ethanol and hexane, and a mixture of an aqueous solution and acetone or ACN.

[0052] For example, it is reported that WO 2021 / 129589 A1 identifies solvents that are purported to be "good solvents" for dissolving besudil to produce its amorphous form. WO 2021 / 129589 A1 provides a working example (Example 22 thereof) relating to the preparation of amorphous KD025, the preparation comprising using DMF as a solvent. However, the applicant has found that besudil is only slightly soluble in DMF, and thus DMF is not a suitable solvent. WO 2021 / 129589 A1 further identifies solvents recommended for the preparation of amorphous besudil, purportedly selected from one or more of methanol, acetone, methyl ethyl ketone, DMF, dimethyl sulfoxide (DMSO), n-methylpyrrolidone, and ethylene glycol dimethyl ether. However, WO 2021 / 129589 A1 does not provide working examples describing the use of these solvents. The applicant has found through actual working examples as described in the present application that the solvents identified in WO 2021 / 129589 A1 are not suitable solvents as defined in the present application due to the low solubility of besudil and / or the carrier matrix in the solvents and / or their incompatibility during spray drying and / or the drug development process.

[0053] Amorphous solid dispersions comprising besudil can be used to prepare solid pharmaceutical dosage forms such as tablets and capsules.

[0054] In one aspect, the present application provides a pharmaceutically acceptable composition comprising a therapeutically effective amount of amorphous besuldiol formulated with one or more pharmaceutical excipients. The pharmaceutical composition can be specifically formulated for administration in solid form and adapted for administration to a patient via a method compliant with using a solid form of the API, such as via oral administration, for example, administration with tablets or capsules.

[0055] In some embodiments, the present application provides a solid pharmaceutical dosage form for oral administration (capsules, tablets, pills, powders, granules, etc.) comprising an amorphous solid dispersion of besuldiol, which is mixed with one or more pharmaceutically acceptable excipients (including pharmaceutically acceptable carriers such as sodium citrate or dicalcium hydrogen phosphate) and / or any of the following: (1) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and / or basic silicate; (2) binders such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and / or gum arabic; (3) wetting agents such as glycerol; (4) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) dissolution retardants such as paraffin wax; (6) absorption accelerators such as quaternary ammonium compounds and surfactants such as poloxamer and sodium dodecyl sulfate; (7) wetting agents such as cetyl alcohol, glyceryl monostearate, and nonionic surfactants; (8) absorbents such as kaolin and bentonite; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, zinc stearate, sodium stearate, stearic acid, and mixtures thereof; (10) coloring agents; and (11) controlled release agents such as crosslinked polyvinylpyrrolidone or ethyl cellulose. In the case of capsules, tablets, and pills, the pharmaceutical composition may also contain buffering agents. Similar types of solid compositions can also be used as fillers in soft and hard shell gelatin capsules with such excipients as lactose or milk sugar and high molecular weight polyethylene glycols, etc.

[0056] Tablets can be prepared by compression or molding (including melt extrusion), optionally with one or more auxiliary ingredients. Compressed tablets can be prepared using binders (such as gelatin or hypromellose), lubricants, inert diluents, preservatives, disintegrants (such as sodium starch glycolate or croscarmellose sodium), surfactants, or dispersants. Molded tablets can be prepared by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0057] Tablets and other solid dosage forms (such as capsules, pills, and granules) of the pharmaceutical composition of the present application can optionally be scored or prepared with coatings and shells (such as enteric coatings and other coatings well-known in the field of pharmaceutical formulation). They can also be formulated to provide slow or controlled release of the active ingredient, for example, using different proportions of hypromellose, other polymer matrices, liposomes, and / or microspheres that provide the desired release profile. They can be formulated for rapid release, such as by lyophilization. These compositions can also optionally contain opacifying agents and can have a composition such that they release only one or more active ingredients, or optionally, release in a delayed manner in a certain part of the gastrointestinal tract. Examples of embedding compositions that can be used include polymeric substances and waxes. If appropriate, the active ingredient can also be in the form of microcapsules with one or more of the above excipients.

[0058] In addition to inert diluents, oral compositions can also contain adjuvants such as wetting agents, emulsifying agents, and suspending agents, sweetening agents, flavoring agents, coloring agents, fragrances, and preservatives.

[0059] A pharmaceutical composition containing an effective amount of amorphous besudil can be used to inhibit ROCK1 and ROCK2, preferentially inhibit ROCK2, and thus can be used to treat diseases regulated by ROCK enzymes, such as autoimmune diseases and / or fibrotic diseases, especially including GVHD (chronic and acute), pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, radiation fibrosis, or arterial fibrosis, cardiac fibrosis, endomyocardial fibrosis, renal fibrosis, or liver fibrosis; moderate to severe psoriasis, rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus (SLE), Crohn's disease, dermatitis (e.g., atopic dermatitis), and eczema and other indications.

[0060] The composition provided by the present application can further be used to treat bronchiolitis obliterans syndrome (BOS), which is a potentially serious complication after lung transplantation or allogeneic hematopoietic stem cell transplantation (allogeneic HSCT). Examples

[0061] In view of the disclosure of the present application, the following abbreviations can be used for reference. Abbreviations: ACN Acetonitrile API Active Pharmaceutical Ingredient (in this application, KD025) DCM Dichloromethane DSC / TGA Differential Scanning Calorimetry / Thermogravimetric Analysis FaSSIF Fasted State Simulated Intestinal Fluid GC - HS Headspace Gas Chromatography HPLC High Performance Liquid Chromatography HPMC Hydroxypropyl Methylcellulose HPMCAS Hydroxypropyl Methylcellulose Acetate Succinate h / hr hour MDSC Modulated Differential Scanning Calorimetry MEK Methyl Ethyl Ketone PK Pharmacokinetics PLM Polarizing Light Microscopy PMA Polymethacrylate PO Per Oral; Oral Administration PPPEG Polyvinylcaprolactam - Polyvinylacetate - Polyethylene Glycol PSD Particle Size Distribution PVP Polyvinylpyrrolidone PVPVA Polyvinylpyrrolidone - Vinyl Acetate RCS Mechanical Refrigeration System RH Relative Humidity RRT Relative Retention Time RT Room Temperature SDD Spray - Dried Dispersion SEM Scanning Electron Microscopy TEA Triethylamine Temp. Temperature THF Tetrahydrofuran TFA Trifluoroacetic Acid XRPD X - Ray Powder Diffraction Instruments:

[0062] Unless other instrument details are set forth in the following examples, the following instruments and procedures can be used to collect the data set forth in the examples of the present application. Those skilled in the art can understand that alternative instruments and procedures can optionally be used to collect characterization data such as PLM, XRPD, TGA, DSC / TGA, and PSD.

[0063] Collect XRPD, PSD, and MDSC data using the instruments and procedures described in Table 1, Table 2, and Table 3 below, respectively: Table 1 Table 2 Table 3 Example 1 1.1 Solvent Screening

[0064] The purpose of the experiments described in this example was to explore amorphous solid dispersions using spray drying technology to enhance the solubility of besuldiol and improve the biological properties of besuldiol. However, spray drying technology requires a suitable solvent system that is compatible with besuldiol and the polymer matrix material.

[0065] To study the suitable solvent system for use with besuldiol and the polymer matrix material, the organic solvent systems described in this section (and in Table 4A, Table 4B, and Table 4C) were prepared and evaluated. A. Solubility Evaluation Using HPLC

[0066] The solubility (w / v) of besuldiol in various solvents was determined using HPLC. As described in Table 4A below, in a 5 mL amber vial, a saturated solution of besuldiol was prepared with approximately 99 - 105 mg of besuldiol and 2.0 mL of solvent. The solution was shaken well and loaded onto a laboratory rotator and rotated at a constant speed of 200 rpm for 24 h. The resulting sample was filtered through a 0.45 μm filter, and the filtrate was used to quantify besuldiol by HPLC using a linear method. Samples of IPA, acetonitrile, ethyl acetate, DCM, toluene, MIBK, acetone, n - heptane, and methyl tert - butyl ether (MTBE) were injected into the HPLC without any further dilution; samples containing methanol, DMF, and DMSO were further diluted and then injected into the HPLC; for the methanol, DMF, and DMSO solutions, 0.1 mL of the filtered solution was transferred to a 100 mL volumetric flask and diluted to volume with diluent and then injected. The solubility is reported in mg / mL in Table 4A. Table 4A: Results of Solubility Experiments Using HPLC B. pH - Based Solubility Evaluation

[0067] The solubility of besuldiol was examined in various pH buffers (i.e., buffers at pH 1.2, 3.5, 4.5, 6.8, 7.4, and 10). To prepare the buffers, the following stock solutions for pH adjustment were prepared: A 0.2 M HCl solution was prepared by transferring 17.0 mL of 35% HCl to 500 mL of water, mixing well, and diluting the solution to 1000 mL with water; a 2 M acetic acid solution was prepared by transferring 116.0 mL of acetic acid to 500 mL of water, mixing well, and diluting to 1000 mL with water; a 0.2 M NaOH solution was prepared by transferring 4.01492 g of sodium hydroxide pellets to 250 mL of water, dissolving, and diluting to 500 mL with water; a phosphate buffer stock solution was prepared by transferring approximately 2.72 g of dipotassium hydrogen phosphate buffer to a 100 mL volumetric flask, dissolving with water, and diluting to the volume.

[0068] Buffers containing the sample (besuldiol) were prepared as follows.

[0069] pH 1.2 buffer of the sample. Transfer 1.53145 g of potassium chloride to a 100 mL volumetric flask, dissolve with water, and dilute to volume. Transfer 25.0 mL of this solution to a 100 mL volumetric flask; add 42.5 mL of 0.2 M hydrochloric acid solution, and dilute the solution to volume with water. Add 2.0 mL of this buffer at pH 1.2 to a 5 mL amber vial containing 100.15 mg of besuldiol.

[0070] pH 3.5 buffer of the sample. Transfer 4.11871 g of potassium hydrogen phthalate to a 100 mL volumetric flask, dissolve with water, and dilute to volume. Transfer 25.0 mL of this solution to a 100 mL volumetric flask, add 8.3 mL of 0.2 M hydrochloric acid thereto, and dilute the solution to volume with water. Add 2.0 mL of this buffer to a 5 mL amber vial containing 100.52 mg of besuldiol.

[0071] pH 4.5 buffer of the sample. Add 25.0 mL of the pH 3.5 buffer (from the previous paragraph) to a 100 mL volumetric flask. Add an additional 10 mL of 0.2 M sodium hydroxide, and dilute the solution to volume with water. Add 2.0 mL of this buffer at pH 4.5 to a 5 mL amber vial containing 101.53 mg of besuldiol.

[0072] pH 6.8 buffer of the sample. Transfer 25.0 mL of the phosphate buffer stock solution to a 100 mL volumetric flask; add 23 mL of 0.2 M sodium hydroxide, and dilute the solution to volume with water. Add 2.0 mL of this buffer at pH 6.8 to a 5 mL amber vial containing 100.18 mg of besuldiol.

[0073] pH 7.4 buffer of the sample. Transfer 25.0 mL of the phosphate buffer stock solution to a 100 mL volumetric flask; add 41 mL of 0.2 M sodium hydroxide, and dilute the solution with water to volume. Add 2.0 mL of this pH 7.4 buffer to a 5 mL amber vial containing 100.52 mg of besuldiol.

[0074] pH 10.0 buffer of the sample. Add 1.52493 g of potassium chloride and 1.21879 g of boric acid to a 100 mL volumetric flask, dissolve with water and dilute to volume. Transfer 25.0 mL of this solution to a 100 mL volumetric flask; add 22 mL of 0.2 M sodium hydroxide, and dilute the solution with water to volume. Add 2.0 mL of this pH 10.0 buffer to a 5 mL amber vial containing 103.8 mg of besuldiol.

[0075] Vigorously shake each of the pH buffers of the samples prepared according to the preceding paragraphs, and load them onto a laboratory spinner and spin at a constant speed of 200 rpm for 24 h. In each case, undissolved samples were observed in the vials after 24 h. Filter the resulting sample solutions through a 0.45 μm needle filter, collect the filtrates, and inject them into the HPLC without any further dilution. Observe the pH after equilibration at 25 °C for 24 hours. The results are shown in Table 4B. Table 4B: Solubility of besuldiol in pH buffers pH of the buffer used pH observed after 24 h of equilibration at 25 °C Solubility (mg / mL) Results 1.2 1.42 NA Very Slightly Soluble 3.5 3.22 NA Very Slightly Soluble 4.5 4.32 NA Very Slightly Soluble 6.8 4.45 NA Very Slightly Soluble 7.4 4.84 NA Very Slightly Soluble 10 5.64 NA Very Slightly Soluble C. Solubility evaluation - Visual observation

[0076] Further conduct solubility evaluations using each of the solvent systems listed in Table 4C, with 25 mg of besuldiol used for each sample. Table 4C: Solvent systems used in the solubility experiments Solvent System Acetone Tetrahydrofuran (THF) Methanol Dichloromethane (DCM) Methyl Ethyl Ketone (MEK) 90:10 DCM:Methanol 80:20 DCM:Methanol 50:50 DCM:Methanol 95:5 Acetone:Water Acetonitrile (ACN) 90:10 ACN:Water 50:50 Ethanol:Hexane

[0077] All solutions were prepared with 10 wt.% besuldiol and then diluted to 1 wt.% besuldiol. Under those conditions, besuldiol was insoluble in any of the solvent systems listed in Table 4C. Then all solutions were heated to 40 °C, but no improvement was observed upon heating. Therefore, the solvent systems listed in Table 4C are not suitable for dissolving besuldiol and the polymer carrier matrix material, and thus the technology cannot be used to prepare the amorphous form. After further experiments, a mixture of besuldiol, triethylamine (TEA) in a 3:1 ratio, and acetone (molar equivalent of TEA:acetone) provided a clear solution of 5 wt.% besuldiol. Therefore, the mixture of TEA and acetone was designated as the solvent system for formulating the solid dispersions used in the following examples. 1.2 Polymer Carrier Matrix and Ratio Selection

[0078] Three polymers were selected to prepare six formulations: (1) Vinylpyrrolidone - Vinyl acetate copolymer (PVPVA) (sold under the trade name VA 64); (2) Polyvinylcaprolactam - Polyvinyl acetate - Polyethylene glycol graft copolymer (PCL - PVAc - PEG, or "PPPEG" in this application) (sold under the trade name and available from BASF); and (3) Hydroxypropylmethylcellulose acetate succinate (HPMCAS - M, or HPMC) (available from Shin Etsu Chemical Co.).

[0079] The ratio of besudil to the polymer carrier matrix (by weight) was selected as follows for evaluation: Table 5: Besudil (KD025): Polymer ratios of six formulations 1.3 Preparation of Spray - Dried Dispersions

[0080] First, a suspension of besudil and TEA in acetone was prepared, and then the three polymer carrier matrix materials in Table 5 were added using the besudil: polymer ratios listed in Table 5. Then the suspension was spray - dried as follows.

[0081] A Buchi B - 290 spray dryer was used in this study. Nitrogen was used as the drying gas. The solution feed rate (mL / min) and atomization pressure (psi) were adjusted to 17.5 and 26 respectively. The inlet temperature ranged between 84 °C - 101 °C, and the outlet temperature was adjusted to a range between 49 °C - 51 °C. The spray - dried formulations were oven - dried at 50 °C for 24 h using a convection disk dryer to remove residual solvents. Example 2

[0082] As described in this Example 2, the surface morphology, crystallinity, and dissolution of all six formulations of Example 1 were characterized. All six formulations produced solid products, which were considered acceptable for further evaluation based on their morphology and amorphous state. However, after analyzing and characterizing the surface morphology, crystallinity, and dissolution as described in Sections 2.1 - 2.3 below, Example 1.2 (20:80 KD025:PVPVA), Example 1.3 (20:80 KD025:PPPEG), and Example 1.6 (40:60 KD025:PPPEG) were selected as preferred candidates for further evaluation as described in Examples 3 - 6. 2.1 Scanning electron microscopy

[0083] The dispersions of Examples 1.1, 1.2, and 1.3 were selected for visual observation using a scanning electron microscope (SEM). Samples were prepared as follows: The samples were dispensed onto adhesive carbon-coated sample stubs and a thin gold conductive layer was coated thereon using a Cressington 108 Auto. The samples were analyzed using a FEI Quanta 200 SEM equipped with an Everhart-Thornley (secondary electron) detector operating in high vacuum mode. The results are shown in Figure 1 this application. Characteristic morphological features of the solid dispersions were observed, which consisted of intact and collapsed spheres with smooth surfaces. However, based on this assessment, the use of a polymeric carrier matrix containing PVPVA and PPPEG was more advantageous than HPMC, since the dispersion containing HPMC showed some filaments, while the dispersion containing PVPVA and PPPEG produced a more homogeneous matrix with besulifloxacin suspended in the polymeric matrix. 2.2 Powder X-ray diffraction

[0084] The crystalline forms of the six dispersions of Example 1 were evaluated using powder X-ray diffraction (XRPD) (using a Rigaku Miniflex 6G X-ray diffractometer). The samples were irradiated with monochromated Cu Kα radiation and analyzed in continuous scan mode between 5° and 40°. The samples were rotated during the analysis to minimize the preferred orientation effect. Figure 2 The XRPD results for each of Examples 1.1 to 1.6 are shown. These results confirm that each of the solid dispersions of Example 1 exists in an amorphous state, as reflected by the absence of crystalline peaks. 2.3 Dissolution assessment

[0085] The initial step in the dissolution assessment was to directly determine the solubility of besulifloxacin in a biorelevant medium (0.1 N HCl or FaSSIF), which was found to be >1000 μg / mL and 5 μg / mL, respectively. The dissolution performance of the six dispersions of Example 1 and crystalline besulifloxacin mesylate was evaluated by non-sink dissolution tests (results are shown in Figure 3Dissolution testing was used to measure this solubility enhancement that is higher than that of bulk crystalline besulindac in a biorelevant FaSSIF medium after 30 minutes of exposure to a low pH environment. During the test, the sample was transferred from 0.1N HCl [theoretical Cmax = 1000 μg A / mL] to FaSSIF [theoretical Cmax = 500 μg A / mL] by dilution. The drug concentration measured in this test is a composite of the SDI free drug, the drug in micelles, and the drug in the drug-polymer colloid. The purpose of this experiment was to rank and select lead formulations. Table 6 summarizes the overall two-stage dissolution data. Table 6: Two-stage dissolution data

[0086] Overall, for most formulations, the Cmax in the low pH of the gastric environment is much higher than that in the high pH. However, as shown in Table 6, the Cmax values of Example 1.3 and 1.6 (two PPPEG dispersions, 20:80 and 40:60) and Example 1.2 (20:80 KD025:PVPVA dispersion) are 369.6 μg A / mL, 79.7 μg A / mL, and 99.9 μg A / mL respectively, which are superior to the other three formulations. In addition, compared with crystalline KD025, the drug concentrations of these formulations at the end of the dissolution process (210 minutes) are higher, with reported values of 20.8 μg A / mL (Example 1.2 [20:80 KD025:PVPVA]), 369.6 μg A / mL (Example 1.3 [20:80 KD025:PPPEG]), 50.4 μg A / mL (Example 1.6 [40:60 KD025:PPPEG]), compared with 6.1 for crystalline KD025. In summary, an increase in the area under the curve (AUC) was observed for Example 1.2 (20:80 KD025:PVPVA), Example 1.3 (20:80 KD025:PPPEG), and Example 1.6 (40:60 KD025:PPPEG) compared to besulindac (KD025). Example 3

[0087] Based on the surface morphology, crystallinity, and dissolution studies of Example 2, three formulations were selected for additional characterization and in vivo evaluation and experiments, namely: Example 1.2 (20:80 KD025:PVPVA), Example 1.3 (20:80 KD025:PPPEG), and Example 1.6 (40:60 KD025:PPPEG). For ease of reference, these selected formulations are hereinafter identified as: Formulation 1 [F1]: 20:80 KD025:PPPEG (Example 1.3); Formulation 2 [F2]: 20:80 KD025:PVPVA (Example 1.2); and Formulation 3 [F3]: 40:60 KD025:PPPEG (Example 1.6).

[0088] For this example, F1, F2, and F3 were prepared according to the processing conditions listed in Table 7 below, and then their amorphous state, surface morphology, residual solvents, particle size distribution, and thermal evaluation were analyzed. Table 7: Methods for Preparing Amorphous Dispersion Formulations F1, F2, and F3 3.1 Amorphous Dispersion

[0089] Using the instrument conditions described in Table 1 above, XRPD results of the three formulations prepared according to the process described in Table 7 were obtained. The results are shown in Figure 5 . It was confirmed that all solid dispersions were amorphous ( Figure 5 ). SEM images of F1, F2, and F3 were also obtained at magnifications of 1500x and 5000x (according to the process in Example 2.1), and the results are shown in Figure 6 . No crystals were observed in any of the three solid dispersions. Formulations F1 and F2 (20:80 dispersions) were spheres with both swelling and collapse, and F3 formed swollen spheres mostly fused in clusters. 3.2 Residual Solvent Content Evaluation

[0090] After the secondary drying step listed in Table 7, GC-HS was used to measure the residual acetone and TEA remaining in these three solid dispersions (F1, F2, and F3). The measurement was carried out using an HP 6890 series GC equipped with an Agilent 7697A headspace sampler. A 30m x 0.32mm x 1.8μm capillary column with a 6% cyanopropylphenyl, 94% dimethylpolysiloxane GC column was used for the test.

[0091] The residual solvents detected in all the formulations are reported in Table 8 below. For all three formulations (F1, F2, and F3), the acetone levels were not detected. For F1 and F3 (PPPEG formulations), the TEA levels were below the limit of quantification (LOQ), and for F2 (PVPVA formulation), the TEA level was 1631 ppm. These levels are below the acetone and TEA limits (5000 ppm) as elucidated by the International Conference on Harmonization (ICH). Table 8: Residual Solvent Evaluation of F1, F2, and F3 Formulation Residual Acetone (ppm) Residual TEA (ppm) Measured Tg °C F1 ND <LOQ 73 F2 ND 1631 90 F3 ND <LOQ 67 ND = Not Detected 3.3 Particle Size Distribution

[0092] The particle size distribution (PSD) of F1, F2, and F3 was measured using the parameters summarized in Table 2 and the laser diffraction method (Mastersizer 3000 with Aero S device). A 200 mg sample was added to a standard venture disperser with a hopper gap of 1.5 mm and then fed into the dispersion system. The feed rate (20% - 40%) was adjusted to keep the laser extinction level between 0.1% and 15%. The sample particles were transported and suspended through the optical cell using compressed air at 1.5 bar. A measurement time of 10 seconds was used, and a background measurement was taken for 10 seconds using air. The Dv10, Dv50, and Dv90 diameters were used to characterize the particle size distribution of the powder.

[0093] The results of the PSD study are shown in Table 9 and Figure 7 in. F1 and F2 (20:80 besuldiol dispersions) showed very similar particle sizes, with Dv50 less than 10 μm, and the particle size ranges of both formulations were quite narrow (the Dv10 - Dv90 values for F1 were approximately 3 to approximately 28, and the Dv10 - Dv90 values for F2 were approximately 2 to approximately 17). F3 (40% besuldiol dispersion) was found to have a bimodal distribution, with larger total particles (Dv10 = 6.54, Dv90 = 187). These results are consistent with the observations obtained by SEM( Figure 6 ), which showed that the particles of F3 were fused together in clusters. Table 9: Particle Size Data of F1, F2, and F3 Formulation Dx(10) (μm) Dx(50) (μm) Dx(90) (μm) F1 3.01 8.32 27.9 F2 1.88 4.56 17.2 F3 6.54 32.9 187 3.4 Differential Scanning Calorimetry

[0094] Perform DSC on F1, F2, and F3 using the instruments listed in Table 3 above. Place the samples in unsealed aluminum pans and heat them at a constant rate of 2.0 °C / min over a temperature range of 0 °C to 240 °C. Purge the system with a nitrogen gas flow of 50 mL / min to ensure an inert atmosphere throughout the measurement.

[0095] The results of this thermal analysis (mDSC) are shown in Figure 8 . All three formulations, F1, F2, and F3, have a single Tg (the Tg (°C) of F1, F2, and F3 are 73, 90, and 67, respectively), indicating their good homogeneity. F3 (40% KD025 dispersion) shows a broad Tg with an ill-defined onset, indicating a high molecular mobility of the dispersion during spray drying. Example 4 4.1 In Vitro Drug Release

[0096] Perform in vitro drug release evaluation of F1, F2, and F3 using a USP II Type Distek 2100 dissolution apparatus. Conduct a two-stage dissolution test. Briefly suspend the pre-weighed SDI powder in the medium and transfer it to 50 mL of pre-heated (37 °C) volume of 0.1 N simulated gastric fluid (SGF) (pH approximately 1.0, without pepsin or bile salts) with a stirrer speed of 100 rpm. After 30 minutes of gastric pH exposure, add 2x concentrated (FaSSIF) to the SGF, resulting in a final pH of 6.8 in FaSSIF (100 mM PBS containing 2.24 mg / mL SIF powder (original) (Biorelevant Inc)) with a total volume of 100 mL. Take 1.0 mL samples at predetermined time points and analyze them using a suitable HPLC method.

[0097] The results are shown in Table 10 and Figure 9 As Figure 9 reflected, the in vitro performance results indicate that F1, F2, and F3 exhibit improved dissolution performance compared to the crystalline mesylate of besudil. The data suggest that F1, F2, and F3 are viable options for achieving higher relative solubility while maintaining acceptable chemical and physical stability. Table 10: Non-sink dissolution data of F1, F2, F3 4.2 Assay Evaluation

[0098] Reverse-phase high-performance liquid chromatography (RPHPLC) (Agilent 1200 series LC (1220 and 1260)) was used to determine and analyze the impurities in F1, F2, and F3 during processing compared to besuldiol in mesylate form. The HPLC was equipped with a diode array detector. A gradient method using a Zorbax SB-CN column was used. The mobile phase consisted of (A) 50 mM potassium phosphate buffer and (B) acetonitrile, pumped at a flow rate of 1.4 mL / min at ambient temperature, and the detection wavelength was 250 nm. The mobile phase gradient was maintained as follows (minutes, % B): (0, 20.0); (20.0, 30.0); (30.0, 60); (40.0, 60).

[0099] The results of this determination are reported in Table 11 below and shown in Figure 10 . The impurity profile was similar to that of besuldiol in mesylate form. No degradation was observed during processing. Table 11: Determination evaluation and impurity profile Example 5 5.1 Stability assessment

[0100] To evaluate the physical and chemical stability of formulations F1, F2, and F3, the three formulations were aged for up to 8 weeks at 25 °C / 60% relative humidity (RH) in open packaging and at 40 °C / 75% RH in open and closed packaging. The physical and chemical stability of F1, F2, and F3 was evaluated by appearance and XRPD. The XRPD collected is shown in Figure 11A (F1), Figure 11B (F2), and Figure 11C (F3).

[0101] Overall, even with the Figure 11B observed physical changes shown in, F2 (20:80 KD025:PVPVA) still formed hard solids after eight weeks under all conditions, and the dispersion remained amorphous under all conditions. As shown in Figure 11A , F1 (20:80 PPPEG dispersion) remained amorphous at 25 °C / 60% RH and open conditions, as well as at 40 °C / 75% RH and closed conditions. At 40 °C / 75% RH and open conditions, F1 formed hard lumps. As shown in Figure 11C , F3 remained amorphous at 40 °C / 75% RH and closed conditions and formed crystals at 25 °C / 60% RH and open conditions, as well as at 40 °C / 75% RH and open conditions. This study provides information on the time and storage conditions required for pre-manufacture of the final dosage form (i.e., tablets, suspensions, etc.). Example 6

[0102] As described in this Example 6, after oral (PO) administration, the pharmacokinetics (PK) of F1, F2, and F3 were evaluated in male beagle dogs. 6.1 Suspension Formulations for In Vivo Administration

[0103] For the PK dog model, suspension formulations for administering F1, F2, and F3 were developed. Suspensions of 25 mg A / mL were prepared with each of F1, F2, and F3 in 0.5 wt.% Methocel A4M, and the visual appearance, injectability, and crystallinity were evaluated by PLM. Methylcellulose A4M was added to purified preheated water (65 °C ± 5 °C) until it was completely dispersed in the water. Then, with continuous mixing, the mixture was cooled to room temperature. Based on the required dosing scenario, the amount of the solid dispersion (F1, F2, F3) powder was added slowly. Initially, a wet paste was formed and it turned into a suspension as mixing continued. The suspensions were evaluated by PLM (5X magnification) at T = 0, T = 1 h, and T = 2.5 h; the images are shown in Figure 12 in.

[0104] The F1 dispersion (20:80 KD025:PPPEG) was homogeneous, without agglomerates or crystals, and remained unchanged within 2.5 hours. At room temperature with stirring, the F1 formulation remained stable in suspension form for at least 4.5 h. Both F2 (20:80 KD025:PVPVA) and F3 (40:60 KD025:PPPEG) showed agglomerates by PLM at T = 0, and the agglomerates grew over time; however, no crystals were observed and the suspensions remained injectable (injected through a 20-gauge gavage tube).

[0105] The suspension formulations were successfully developed and selected for ease of administration to the dog PK model; however, solid dosage forms can also be considered. Solid dosage forms (such as tablets) are relatively easy to manufacture, package, and transport, are more stable than liquids, and can be formulated and shaped with coatings to facilitate swallowing. Therefore, those skilled in the art can consider using solid dosage forms as an alternative to the liquid suspensions used in this dog study. 6.2 In Vivo Canine PK Evaluation

[0106] Male beagle dogs were selected for the biological performance evaluation of F1, F2, and F3. All studies were conducted according to protocols approved by the Pharmaron Institutional Animal Care and Use Committee. Twenty male beagle dogs, 1 - 1.5 years of age, were assigned dosing and feeding conditions, with their body weights maintained between 11 and 12 kg throughout the course of the study. All animals were housed in an environment with a 12-hour light / dark cycle. F1, F2, and F3 were tested under fasting and fed conditions and compared to besulforhodamine free base powder and immediate release (IR) tablets containing besulforhodamine crystalline mesylate used as controls. For the fasting group, the dogs were fasted overnight and the drug was administered to the dogs simultaneously in the fasting state in the morning. After collecting plasma 4 hours after dosing, the food was returned to the dogs. For the fed group, the dogs were fasted overnight and fed 1 hour before dosing. Throughout the study, all dogs had access to water. All samples were administered orally (PO); the tablets were administered as received. After tablet administration, 5 mL of vehicle was used to assist the dogs in swallowing the tablets. Formulations 1, 2, 3, and the free base sample were administered via oral gavage using a suspension formulation of 0.5 wt% Methocel A4M. After suspension administration, 5 mL of vehicle was given to the dogs to ensure that all of the suspension was flushed into the stomach. 6.3 Study Design

[0107] A total of twenty beagle dogs were dosed according to the dosing regimens summarized in Table 12. Four dogs were assigned to each of groups A - J. Groups A / F, B / G, C / H, D / I, and E / J shared the same animals; the animals in groups A - E were dosed first. After a seven-day washout period, the animals in groups F - J were dosed. In each case, dosing was performed via oral administration. Four male dogs in each group were dosed. Table 12: Beagle Dog PK Dosing Information for F1, F2, and F3 and API Control 6.4 Sample Collection

[0108] Blood samples were collected from each animal at pre-determined time points before dosing and at 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, 18 hours, 24 hours, 30 hours, and 36 hours after dosing. Blood samples (1 mL) were collected from each animal via the jugular vein. These blood samples were placed in tubes containing dipotassium ethylenediaminetetraacetate and then centrifuged at 2000 g for 10 minutes at 2°C to 8°C to obtain plasma. 6.5 LC / MS Conditions

[0109] Evaluation of KD025 in dog plasma samples using an LC-MS / MS system consisting of two Shimadzu LC-30AD pumps, a DGU-20A5R degasser, a Rack changer II, and an AB Sciex Triple Quads 5500 LC / MS / MS mass spectrometer. Chromatographic separation was performed at room temperature on an Agilent ZORBAX XDB-Phenyl 5 μm (50×2.1 mm) column. The mobile phase consisted of the following: A: 5% acetonitrile (0.1% formic acid) in water; B: 95% acetonitrile (0.1% formic acid) in water. The flow rate was 0.6 mL / min. The injection volume was 2 μL, and the lower limit of quantification (LLOQ) was 10 ng / mL. 6.6 Data collection and statistical analysis

[0110] Data acquisition was performed using Sciex Analyst 1.6.3 software (AB Sciex, Foster City, CA). Pharmacokinetic parameters such as area under the curve (AUC0-36h), maximum plasma concentration (Cmax), and time to reach Cmax (Tmax) were calculated by non-compartmental model analysis (Phoenix TM WinNonlin TM 6.1). The AUC was calculated using the linear trapezoidal algorithm. Data statistics and plasma profile analysis were performed using Excel 2010 software.

[0111] Table 13 summarizes the mean pharmacokinetic parameters after administration of a 40 mg / kg dose of the test formulation to male beagle dogs (n = 4). Figure 13 Plots of plasma concentration versus time of KD025 were made for the following dog groups over a twenty-four data collection period: Group A = fasted, tablet formulation; Group D = fasted, F2 formulation (20:80 KD025:PVPVA); Group F = fed, tablet formulation; Group I = fed, F2 (20:80 KD025:PVPVA). Table 13: Mean plasma pharmacokinetic parameters after oral administration at 1000 mg / dog in beagle dogs

[0112] When comparing the solid dispersion formulations F1, F2, and F3 with the tablets, it was surprisingly found that F2 (20:80 KD025:PVPVA) performed better than the other (PPPEG) formulations. For F2, the mean Cmax in the fasting group and the fed group was found to be similar, i.e., the mean Cmax in the fasting group was 2915.0 ng / mL and the mean Cmax in the fed group was 3100 ng / mL. In contrast, the mean Cmax in the fasting state of the reference tablet was significantly different from that in the fed state, i.e., the mean Cmax in the fasting group was 2842 ng / mL and the mean Cmax in the fed group was 4773 ng / mL. From a pharmacokinetic perspective, this improved control of Cmax by amorphous besudil may be beneficial.

[0113] In addition, compared with the reference tablet, the F2 (KD025:PVPVA) test group demonstrated lower variability, as characterized by %CV (100 x standard deviation / mean). Under fasting conditions, the %CV of the F2 formulation was found to be 28.9, while that of the tablet was 83.0. When compared with the fasting condition, the percentage variability of the F2 formulation was even lower, at 7.5% CV, while that of the reference tablet was 32.9% CV. In summary, in terms of Cmax, the solid dispersion formulation F2 (20:80 KD025:PVPVA) reduced inter-subject variability and minimized the food effect. The AUC observed with the F2 formulation was slightly lower than that of the actual reference tablet; however, the variability was better controlled. Under fasting conditions, using %CV indicated that the AUC variability (33.1) of F2 (20:80 KD025:PVPVA) was improved compared with that of the tablet (97.0). In summary, the use of the solid dispersion formulation F2 was unexpectedly effective in achieving minimal variability, and the food effect of F2 was lower than that of the free base tablet. During the study of this example, no side effects on the animals were observed during or after the study.

Claims

1. A solid dispersion comprising substantially amorphous 2-{3-[4-(1H-indazol-5-ylamino)-2-quinazolinyl]phenoxy}-N-(propan-2-yl)acetamide or a pharmaceutically acceptable salt thereof (bezulifloxacin) and one or more carrier materials.

2. The solid dispersion according to claim 1, wherein the one or more carrier materials are selected from polymers.

3. The solid dispersion according to claim 1, wherein the one or more carrier materials are selected from polyvinylpyrrolidone-vinyl acetate copolymer, polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol graft copolymer, and hypromellose acetate succinate.

4. The solid dispersion according to claim 1, 2 or 3, wherein the ratio of bezulifloxacin to the one or more carrier materials is about 10:90 to 90:10 by weight; or about 20:80 to 80:20 by weight; or about 25:75 to 75:25 by weight; or about 40:60 to 60:40 by weight.

5. The solid dispersion according to any one of claims 1 to 4, wherein the carrier material is polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol graft copolymer.

6. The solid dispersion according to any one of claims 1 to 4, wherein the carrier material is polyvinylpyrrolidone-vinyl acetate copolymer.

7. The solid dispersion according to any one of claims 1 to 3 or claim 5 or 6, wherein the ratio of bezulifloxacin to the carrier material is about 20:80 by weight.

8. The solid dispersion according to any one of claims 1 to 7, wherein at least about 95% of bezulifloxacin in the dispersion is in an amorphous form.

9. The solid dispersion according to any one of claims 1 to 7, wherein at least about 99% of bezulifloxacin in the dispersion is in an amorphous form.

10. The solid dispersion according to claim 8 or 9, characterized in that The solid dispersion contains solid particles with a particle size diameter less than 10 μm.

11. A pharmaceutical formulation comprising a therapeutically effective amount of the solid dispersion according to any one of claims 1 to 10.

12. The pharmaceutical formulation according to claim 11, wherein the pharmaceutical formulation is a tablet or a capsule.

13. A method for treating a disease or disorder regulated by ROCK, the method comprising administering to a subject in need of treatment the solid dispersion according to any one of claims 1 to 10 or the pharmaceutical formulation according to claim 11 or 12.

14. The method according to claim 13, wherein the disease or disorder is graft-versus-host disease (GVHD).

15. The method according to claim 14, wherein the GVHD is chronic or acute.

16. The method according to claim 13, wherein the disease or disorder is an autoimmune disease or a fibrotic disease.

17. The method according to claim 16, wherein the autoimmune disease or fibrotic disease is pulmonary fibrosis; idiopathic pulmonary fibrosis; cystic fibrosis; radiation fibrosis; arterial fibrosis, cardiac fibrosis, endomyocardial fibrosis, renal fibrosis or hepatic fibrosis; moderate to severe psoriasis; rheumatoid arthritis; multiple sclerosis; systemic lupus erythematosus (SLE); Crohn's disease; dermatitis; or eczema.

18. The method according to claim 13, wherein the disease or disorder is bronchiolitis obliterans syndrome (BOS).

19. The method according to claim 18, wherein the BOS is BOS after lung transplantation or BOS after allogeneic hematopoietic stem cell transplantation (allogeneic HSCT).

20. A method for preparing amorphous besulforide, the method comprising dissolving besulforide and one or more carrier materials in a suitable solvent to form a solution.

21. The method according to claim 20, wherein the suitable solvent comprises a mixture of triethylamine and acetone.

22. The method according to claim 20 or 21, the method further comprising spray drying the solution of besulforide and one or more carrier materials to remove the suitable solvent.

23. The method according to any one of claims 20 to 22, wherein the carrier material is selected from vinylpyrrolidone-vinyl acetate copolymer and polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol graft copolymer.

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