Composition

Abstract

To provide a compound having specific utility in treatment or prevention of a disorder or a clinical condition improved by AMPK activation.SOLUTION: Provided are alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide expressed by the following structural formula, and a preparation thereof.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] The present invention relates to alkali metal salts of certain pharmaceutically active ingredients and the use of such salts in pharmaceutical preparations. In particular, the present invention relates to oral dosage forms comprising the alkali metal salt of the active ingredient in a finely divided form. [Background technology]

[0002] AMP-activated protein kinase (AMPK) is a protein kinase enzyme composed of three protein subunits. It is activated by hormones, cytokines, exercise, and stressors that reduce cellular energy status (e.g., glucose deprivation). AMPK activation increases processes that generate adenosine 5'-triphosphate (ATP) (e.g., fatty acid oxidation) and limits other processes that consume ATP but are not urgently required for survival, such as fatty acid synthesis, glycerolipid synthesis, and protein synthesis. Conversely, when cells are persistently exposed to excess glucose, AMPK activity decreases, enhancing fatty acid synthesis, glycerolipid synthesis, and protein synthesis. Thus, AMPK is a protein kinase enzyme that plays a critical role in cellular energy homeostasis. Thus, AMPK activation is linked to glucose-lowering effects and triggers several other biological effects, including the inhibition of cholesterol synthesis, lipogenesis, triglyceride synthesis, and the reduction of hyperinsulinemia.

[0003] In view of the above, AMPK is a preferred target for the treatment of metabolic syndrome, particularly type 2 diabetes. AMPK is also involved in many pathways important in many different diseases (e.g., AMPK is also involved in many pathways important in CNS disorders, fibrosis, osteoporosis, heart failure, and sexual dysfunction).

[0004] AMPK is also involved in many pathways important in cancer. Several tumor suppressors are part of the AMPK pathway. AMPK functions as a negative regulator of the mammalian TOR (mTOR) and EF2 pathways, which are key regulators of cell growth and proliferation. Therefore, deregulation may be associated with diseases such as cancer (as well as diabetes). Therefore, AMPK activators may be useful as anticancer drugs.

[0005] 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide (i.e., the compound of formula I) was first disclosed in WO2011 / 004162 and has been shown to be an AMPK activator.

[0006] [ka]

[0007] As AMPK agonists (i.e., AMPK activators), the compounds of Formula I are useful in treating disorders or conditions ameliorated by activation of AMPK. Such compounds may be useful in treating cancer, diabetes, cardiovascular disease, hyperinsulinemia and related conditions, conditions / disorders in which fibrosis plays a role, sexual dysfunction, osteoporosis, and neurodegenerative diseases. Summary of the Invention

[0008] There remains a need to improve the in vivo bioavailability of active ingredients such as 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide in order to improve their effectiveness in medicine. The present inventors have surprisingly found that the use of a specific salt of the compound of formula I improves the bioavailability of the compound in vivo.

[0009] The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge.

[0010] According to a first aspect of the present invention, there is provided an alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide.

[0011] An "alkali metal" is a metal that, along with hydrogen, is included in Group I of the periodic table. The alkali metals are lithium, sodium, potassium, rubidium, cesium, and francium. An "alkali metal salt," therefore, is understood to be a compound consisting of a collection of one or more alkali metal cations and associated anions.

[0012] Thus, the term "alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide" refers to a compound comprising an alkali metal cation (e.g., lithium, rubidium, cesium, and, particularly, sodium and potassium) and an anion of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. For example, "the alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide may be referred to as a compound of formula II:

[0013] [ka] In the formula, X + represents an alkali metal (e.g., lithium, rubidium, cesium, or, especially, sodium or potassium) cation.

[0014] Those skilled in the art will recognize that when dissolved in a suitable solvent (e.g., water), the alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can dissociate into its anionic and cationic components.

[0015] The compound name 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide was derived using the commercially available software package Autonom (a brand of naming software provided as an add-on for use with the Symyx Draw 2.1™ office suite sold by MDL Information Systems).

[0016] Throughout this specification, structures may or may not be displayed with a chemical name. In the event of any doubt regarding nomenclature, the structure shall prevail. Where a compound may exist as a tautomer (e.g., in an alternative resonance form), the depicted structure represents one of the possible tautomeric forms, and the actual tautomeric form observed may vary depending on environmental factors such as solvent, temperature, or pH. All tautomeric (and resonance) forms and mixtures thereof are included within the scope of the present invention. For example, the following tautomers are included within the scope of the present invention:

[0017] [ka]

[0018] For the avoidance of doubt, the alkali metal salts of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide are solids under ambient conditions and therefore the scope of the present invention includes all amorphous, crystalline and partially crystalline forms thereof.

[0019] Alkali metal salts of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be prepared according to techniques well known to those skilled in the art. For example, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be reacted with the appropriate alkali metal hydroxide or an alternative alkali metal base compound. Salt switching techniques can also be used to convert one salt to another.

[0020] When the product is prepared from 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be prepared according to techniques well known to those skilled in the art. For example, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be prepared according to the techniques described in International Patent Application No. 2011 / 004162, the entire contents of which are incorporated herein by reference.

[0021] Unless otherwise defined, all technical and scientific terms used herein have the common meaning as understood by one of ordinary skill in the art to which this invention belongs.

[0022] In a specific embodiment, the alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide is the sodium or potassium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. Preferably, the salt is the sodium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. The sodium salt has been found to exhibit particularly enhanced bioavailability in vivo, as evidenced by data from the examples showing increased plasma exposure when the active ingredient is administered in this form.

[0023] For the avoidance of doubt, those skilled in the art will understand that reference herein to a compound of a particular aspect of the invention (e.g., the first aspect of the invention) includes reference to all embodiments and particular features thereof, and that the embodiments and particular features may be combined to form further embodiments and features of the invention.

[0024] The salts according to the first aspect of the invention are referred to herein as "salts of the invention".

[0025] Pharmaceutical preparations As provided herein, the salts of the present invention are useful as therapeutic agents for treating a variety of medical disorders or conditions. Typically, the salts of the present invention will be administered to a subject in need thereof in the form of a pharmaceutical formulation.

[0026] According to a second aspect of the present invention, there is provided a pharmaceutical formulation comprising an alkali metal salt (e.g., sodium salt) of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. Such a formulation is hereinafter referred to as the "formulation of the present invention." All embodiments and specific features described herein with respect to the first aspect of the present invention are also disclosed herein with respect to the second aspect of the present invention.

[0027] In a particular embodiment, the pharmaceutical formulation comprises a salt as defined in the first aspect of the invention, including all embodiments and particular features thereof, wherein the salt is milled.

[0028] As used herein, the term "comminuted" (which may be used interchangeably with other art terms such as "size-reduced," "comminuted," "ground," and "pulverized") refers to a solid sample that has been subjected to mechanical energy (e.g., by grinding) to reduce the particle size. For example, coarse particles may be broken down into finer particles, thereby reducing the average particle size.

[0029] The phrase "milled salt" will be understood to refer to a salt of the invention (as defined in the first aspect of the invention) obtained by any process which involves milling the salt so that the average particle size of the solid salt product is smaller than the average particle size of the corresponding salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide produced by processes known in the art but which has not been treated to reduce its particle size.

[0030] Milling is considered a "top-down" approach to producing fine particles. For example, drug solids can be cut with sharp blades (e.g., cutter mills), hammered, subjected to high-pressure homogenization, or crushed or compressed by the application of pressure (e.g., roller mills or pestle and mortar). Because a limited amount of energy is usually applied, the particles produced by such methods remain relatively coarse. Technological advances in milling equipment have made it possible to produce ultrafine drug particles down to micron (i.e., μm range) or even submicron (e.g., nm range) dimensions.

[0031] Certain milling processes can be characterized as dry milling processes, and such processes are preferred for treating the salts of the present invention.

[0032] "Dry milling" refers to a process in which a drug is milled in its dry state, i.e., in the absence of a liquid medium (e.g., substantially in the absence of water). In the dry state, the drug can be milled alone or in the presence of one or more other ingredients, such as pharmaceutically acceptable excipients. Other abrasive materials, such as salts, may be present during the milling process to aid in particle size reduction. The mechanical energy imparted by dry milling promotes interactions between particles of the drug (and optionally other substances present) via van der Waals forces or hydrogen bonding.

[0033] A review of pharmaceutical milling processes can be found, for example, in Loh et al., Asian Journal of Pharmaceutical Sciences, 10(2015), 255-274. Suitable excipients for inclusion in drug particles are known in the art and are described, for example, in Peltonen et al., Handbook of Polymers for Pharmaceutical Technologies, ed. Thakur and Thakur, Wiley, volume 4, chapter 3, 67-87, and Nekkanti et al., Drug Nanoparticles—An Overview, The Delivery of Nanoparticles, IntechOpen. The contents of these documents are incorporated by reference.

[0034] The salts and formulations of the invention (as defined by the first and second aspects of the invention, and all embodiments of those aspects) have been found to be surprisingly effective in improving (e.g., increasing) the in vivo bioavailability of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide compared to pharmaceutical formulations comprising 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide in the free base form. The improvement in bioavailability can be measured by measuring the C max Or it can be demonstrated by measuring the area under the curve (AUC). The salts and formulations of the present invention are useful in the therapies described in the present invention in subjects in need of such therapy. Preferably, the subject is a human.

[0035] In the context of the present invention, the term "free base" refers to the form of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide that is not in the form of a salt. For example, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide is not in an ionic (e.g., anionic) form associated with one or more oppositely charged ions (e.g., cations). Free base 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be depicted as a compound of formula I.

[0036] [ka]

[0037] "C maxIt will be appreciated by those skilled in the art that the terms " and "AUC" in the present context refer, respectively, to the peak plasma concentration of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide after administration (e.g., to a human subject) and the integral of the concentration / time curve for that substance after administration of a salt or formulation of the invention.

[0038] It has surprisingly been found that administration of a salt of the active ingredient results in a significantly increased systemic exposure of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide compared to administration of the compound in its non-salt form (i.e., the free form of the compound).

[0039] Stabilizers such as polymers and surfactants are often used during the milling process to increase interparticle repulsion and inhibit aggregation. Finely ground particles may aggregate during pulverization, which ultimately slows the dissolution process and may affect bioavailability. It has been found that increased systemic exposure of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide occurs after administration of a dry-milled salt of the active ingredient, even without the addition of stabilizers. Therefore, in one embodiment, the pharmaceutical formulation does not contain any stabilizers.

[0040] Thus, the formulation of the second aspect of the present invention may increase the bioavailability of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide compared to a pharmaceutical formulation comprising the free base form of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide.

[0041] When we state that "the bioavailability of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be increased compared to a pharmaceutical formulation comprising the free base form of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide," we mean that administration of a formulation comprising a salt of the invention results in a greater systemically available fraction of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide in vivo compared to administration of a formulation comprising free 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. The increase in the amount of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide that is systemically available following administration of a formulation comprising a salt of the invention compared to administration of a formulation comprising free 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be at least about 10%, (at least) about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100% (i.e., 2-fold), about 150%, about 200% (i.e., 3-fold), about 250%, about 300% (i.e., 4-fold), about 350%, or about 400% (i.e., 5-fold).

[0042] The improvement in bioavailability provided by the preparation of the present invention can be demonstrated by using suitable methods known in the art.For example, the improvement in bioavailability can be demonstrated by comparing the pharmacokinetic data (for example, AUC data) of the subject who is administered the preparation that comprises the salt of the present invention with the corresponding data of the subject who is administered the pharmaceutical preparation that comprises 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide in free base form.

[0043] Milling reduces the average size of particles containing the alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. The extent and effectiveness of milling can be determined by measuring the particle size distribution of the particles before and after the milling process.

[0044] The term "particle size distribution" refers to the relative number of particles present according to size in a solid sample, such as a powder, a granular material, or particles dispersed in a fluid. Particle size distribution affects the properties of a solid sample (e.g., a powder) in many ways. The inventors have found that reducing the average particle size leads to a surprising improvement in the bioavailability of the resulting pharmaceutical product.

[0045] The particle size distribution of a solid sample can be measured using techniques well known in the art. For example, the particle size distribution of a solid sample can be measured by laser diffraction, dynamic light scattering, image analysis (e.g., dynamic image analysis), sieve analysis, air sieve analysis, optical counting, electrical resistance counting, sedimentation, laser obscuration, and acoustic (e.g., ultrasonic attenuation) spectroscopy. Specific methods that can be mentioned for measuring the particle size distribution of the salt particles of the present invention are dynamic light scattering and laser diffraction.

[0046] The particle size distribution can also be determined based on the results of sieve analysis. Sieve analysis provides particle size information in the form of an S-curve of the cumulative mass retained on each sieve versus the sieve mesh size. The most commonly used metric when describing particle size distribution is the D-value (e.g., D10, D50, and D90, which are the 10%, 50%, and 90% intercepts of the cumulative mass, respectively). The particle size distribution of the present invention is preferably defined using one or more of such values. The D-value essentially represents the diameter of a sphere that divides the mass of a sample into a specified percentage when the particles are arranged on an ascending mass basis. For example, the D10 value is the diameter at which 10% of the mass of the sample is composed of particles having a diameter less than this value. The D50 value is the diameter of particles that 50% of the mass of the sample are smaller than and 50% of the mass of the sample are larger than.

[0047] Particles comprising a salt of the present invention may have a particle size distribution defined by a D90 of less than about 10 μm (e.g., about 5 μm to about 10 μm). Particles comprised of a salt of the present invention may have a particle size distribution defined by a D50 of less than about 6 μm (e.g., about 1 μm to about 6 μm). The particle size distribution of particles comprised of a salt of the present invention may be further defined by a D10 of less than about 2 μm (e.g., about 0.5 μm to about 2 μm). In certain embodiments, a formulation comprises particles comprising a salt of the present invention, the particles having a particle size distribution defined by a D90 of less than about 10 μm.

[0048] The above particle size distribution parameters may be applicable individually or in combination to any given salt, mixture, or formulation. For example, in certain embodiments, a formulation comprises particles containing a salt of the invention, the particles having a particle size distribution defined by a D90 of less than about 10 μm and a D50 of less than about 6 μm.

[0049] The particle size distribution of particles comprising the salt of the invention can be measured, for example, by laser diffraction using a particle size analyzer such as a Shimadzu SALD-2300. Where such a process involves dispersion of the substance to be analyzed in a liquid medium such as water, a suitable amount of dispersing agent, for example, Tween 20 (polyethylene glycol sorbitan monolaurate), can be used.

[0050] The present invention also includes pharmaceutical formulations comprising particles containing a salt of the present invention having any of the particle size distributions defined herein, regardless of the process by which the particles are produced.

[0051] Preferably, the pharmaceutical formulation of the second aspect of the invention comprises particles of a salt of the invention having any of the particular particle size distributions described herein, the particles being obtained by a process involving milling the salt.

[0052] The pharmaceutical formulations of the second aspect of the invention may be prepared in accordance with standard and / or accepted pharmaceutical practice.

[0053] In an embodiment of the second aspect of the invention, the salt of the invention is the only pharmaceutically active ingredient present in the salt-containing particle. In a further embodiment of the second aspect of the invention, the salt of the invention may be present in a formulation together with one or more other pharmaceutically active ingredients or may be administered as part of a combination therapy with one or more other pharmaceutically active ingredients.

[0054] As described herein, the formulations of the invention of the second aspect may, for example, comprise particles containing a salt of the invention wherein the salt has been milled so that the particles have a particle size distribution defined by a D90 of less than about 10 μm.

[0055] Thus, according to a third aspect of the present invention, there is provided a process for preparing a formulation as defined above (i.e. a formulation according to the second aspect of the present invention, and all embodiments thereof), which process comprises milling a salt of the invention, optionally together with one or more excipients, to produce particles having a particle size distribution defined by a D90 of less than about 10 μm.

[0056] In one embodiment of the third aspect of the present invention, the particles are milled using dry milling. Dry milling typically involves milling a substance (i.e., a drug) in the substantial absence of other components (e.g., in the substantial absence of any other components of a pharmaceutical formulation). Thus, in one embodiment, the particles consist essentially of the salt of the present invention and have a particle size distribution as defined herein. In another embodiment, the particles consist essentially of the salt of the present invention and are obtained by dry milling.

[0057] Optionally, dry milling involves jet milling. Jet milling is a milling process that involves a high-velocity stream of compressed air to reduce the size of drug particles, typically from the 20-100 μm range, to less than 10 μm. Particle size reduction is achieved through a combination of impaction (either between particles or against the walls of the milling chamber) and abrasion (when particles move in opposite directions). Those skilled in the art will understand that particle size control can be achieved by varying milling conditions according to methods known in the art.

[0058] In certain embodiments of the process of the third aspect of the present invention, the salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide is mixed with one or more excipients after grinding the salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide.

[0059] In other embodiments of the process of the third aspect of the present invention, enteric coating can be introduced into the preparation at any time, provided that the salt of the present invention in the obtained preparation is encapsulated in the enteric coating.For example, when the pharmaceutical preparation is provided in the form of a capsule or tablet, the process can further comprise the step of coating the capsule or tablet with an enteric coating before or after (preferably after) the milled salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide is incorporated into the capsule or tablet.Alternatively, the process can further comprise the step of applying an enteric coating to the milled salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide before the particles are incorporated into the capsule or tablet.

[0060] Dry milling can be useful for producing particles that are generally larger in size than those obtained from wet milling. Particular particle size distributions that may be mentioned in this context include those having a D90 of less than about 10 μm, less than about 9 μm, less than about 8 μm, less than about 7 μm, or less than about 6 μm.

[0061] More specifically, particles containing a salt of the invention present in a formulation as defined herein may have a particle size distribution defined by a D90 of about 5 μm, about 5.5 μm, about 6 μm, about 6.5 μm, about 7 μm, about 7.5 μm, about 8 μm, about 8.5 μm, about 9 μm, about 9.5 μm or about 10 μm, which particle size distribution is preferably for particles consisting essentially of a salt of the invention.

[0062] Preferably, particles containing the salts of the invention have a particle size distribution defined by a D50 of less than about 6 μm, less than about 5 μm, less than about 4 μm, less than about 3 μm, or less than about 2 μm.

[0063] The particle size distribution of particles containing the salts of the invention may also be defined by a D50 of about 1 μm, about 1.5 μm, about 2 μm, about 2.5 μm, about 3 μm, about 3.5 μm, about 4 μm, about 4.5 μm, about 5, about 5.5, or about 6 μm.

[0064] Still further, particles comprising the salt of the present invention may have a particle size distribution defined by D10 of less than about 2 μm, less than about 1.5 μm, or less than about 1 μm.

[0065] The particle size distribution of particles containing the salts of the invention may also be defined by a D10 of about 0.5 μm, about 1 μm, about 1.5 μm, or about 2 μm.

[0066] Thus, in certain embodiments of the formulation (i.e. a formulation according to any embodiment of the second aspect of the invention), the particles containing the salt of the invention have a particle size distribution defined by a D90 of less than 9 μm, a D50 of less than 6 μm, a D50 of less than 5 μm, a D10 of less than 2 μm, or a D10 of less than 1.5 μm.

[0067] The formulation of the second aspect of the present invention is generally provided as a mixture containing a salt of the present invention and one or more pharmaceutically acceptable excipients. The one or more pharmaceutically acceptable excipients can be selected in accordance with standard pharmaceutical practice, taking into full consideration the intended route of administration. Such pharmaceutically acceptable excipients are preferably chemically inert to the active compound and preferably have no adverse side effects or toxicity under the conditions of use. Suitable pharmaceutical formulations can be found, for example, in Remington The Science and Practice of Pharmacy, 19th ed., Mack Printing Company, Easton, Pennsylvania (1995). A brief review of drug delivery methods can also be found, for example, in Langer, Science 249, 1527 (1990).

[0068] pH-adjusting excipients have been found to be particularly advantageous in the formulations of the present invention. A pH-adjusting excipient is one that substantially changes the pH of an aqueous solution of the formulation compared to the pH of an aqueous solution of the same formulation without the excipient. A pH-adjusting excipient can increase (or decrease) the pH of an aqueous solution of the formulation (e.g., to a pH of 8 or higher) compared to the pH of an aqueous solution of the same formulation without the excipient. Such excipients can be useful for increasing (i.e., improving) the aqueous solubility and / or stability of a compound of the present invention in a formulation.

[0069] It has been found that basic excipients are useful in the formulations of the present invention in combination with the salts of the present invention, resulting in improved solubility of the compounds. Thus, in certain embodiments of the second aspect of the present invention, at least one pharmaceutically acceptable excipient is a basic excipient.

[0070] As used herein, the term "basic excipient" refers to a pharmaceutically acceptable excipient that increases the microenvironmental pH of a formulation. Adjusting the microenvironmental pH of a formulation has been found to improve the dissolution of the active ingredient in the formulation, which in turn can lead to enhanced oral absorption of the active ingredient. Specific basic excipients that may be mentioned include magnesium oxide, sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate, magnesium carbonate, and calcium carbonate. In certain embodiments, the basic excipient is magnesium oxide.

[0071] Thus, according to certain embodiments, the formulation further comprises at least one pharmaceutically acceptable excipient. In particular, the at least one pharmaceutically acceptable excipient may be a lubricant, binder, filler, surfactant, diluent, anti-adherent, coating, flavoring, coloring, glidant, preservative, sweetener, disintegrant, adsorbent, buffer, antioxidant, chelating agent, dissolution enhancer, dissolution retarder, or wetting agent.

[0072] Specific pharmaceutically acceptable excipients that may be mentioned include mannitol, PVP (polyvinylpyrrolidone) K30, lactose, saccharose, sorbitol, starch, amylopectin, cellulose derivatives, gelatin, or other suitable ingredients, as well as disintegrants and lubricants such as sodium docusate, magnesium stearate, calcium stearate, sodium stearyl fumarate, and polyethylene glycol wax. In preparing pharmaceutical formulations of the salts of the present invention for oral administration, particles containing the salts of the present invention (preferably after milling) can be mixed together or separately with mannitol, PVP (polyvinylpyrrolidone) K30, and sodium docusate. Thus, in certain embodiments, the formulations of the present invention comprise PVP K30, sodium docusate, and mannitol.

[0073] Such mixtures can then be processed into pellets or granules, or compressed into tablets. Thus, pharmaceutical formulations of the present invention include formulations provided in the form of tablets, minitablets, blocks, pellets, particles, granules, or powders for oral administration.

[0074] Those skilled in the art will understand that the preparations described in the present invention can act systemically, and therefore can be administered appropriately by using suitable techniques known to those skilled in the art.The preparations described herein are usually administered orally in pharmaceutically acceptable dosage forms.Therefore, the pharmaceutical preparation of the second aspect of the present invention is preferably an oral pharmaceutical preparation.

[0075] The formulations of the present invention can be prepared for oral administration in the form of capsules. For example, capsules such as soft gelatin capsules can be prepared to contain the salt of the present invention alone or together with a suitable vehicle, such as vegetable oil, fat, etc. Similarly, hard gelatin capsules can contain the salt of the present invention alone or in combination with a solid powder ingredient such as a disaccharide (e.g., lactose or saccharose), a sugar alcohol (e.g., sorbitol or mannitol), a vegetable starch (e.g., potato starch or corn starch), a polysaccharide (e.g., amylopectin or a cellulose derivative), or a gelling agent (e.g., gelatin).

[0076] Thus, pharmaceutical formulations of the present invention include, for example, formulations provided in the form of capsules or tablets for oral administration.

[0077] Preparations intended for oral administration may further comprise an enteric coating to prevent or minimize dissolution or disintegration in the stomach environment. Thus, oral preparations (e.g., capsules or tablets) coated with an enteric coating may provide targeted release of the salt of the present invention in the small intestine. For example, the enteric coating may be present on the surface of the preparation (e.g., on the surface of a tablet or capsule), or each particle containing the salt of the present invention may be coated with an enteric coating. Thus, in certain embodiments, the preparation further comprises an enteric coating.

[0078] It may be desirable to minimize dissolution or disintegration of a capsule or tablet (or the like) in the gastric environment and / or provide targeted release of an active ingredient in the small intestine. Thus, in certain embodiments, an enteric coating is present on the capsule or tablet. For example, the coating may be provided as an outer layer of the capsule or tablet.

[0079] Alternatively, particles containing a salt of the invention can be individually coated with an enteric coating, and the coated particles can be loaded into a capsule or compressed into a tablet. Thus, in certain embodiments, a capsule or tablet contains particles comprising a salt of the invention, each particle coated with an enteric coating.

[0080] The term "enteric coating" refers to a substance (e.g., a polymer) that is incorporated into an oral dosage form (e.g., applied to the surface of a tablet, capsule, particle, or pellet) and inhibits the dissolution or disintegration of the dosage form in the gastric environment. Enteric coatings are stable at the strongly acidic pH typically found in the stomach, but rapidly degrade at the relatively basic pH of the small intestine. Thus, the enteric coating prevents release of the active ingredient in the dosage form until it reaches the small intestine.

[0081] Any enteric coating known to those skilled in the art can be used in the present invention. Specific enteric coating materials that may be mentioned include beeswax, shellac, alkyl cellulose polymer resins (e.g., ethyl cellulose polymer, carboxymethyl ethyl cellulose, or hydroxypropyl methyl cellulose phthalate) or acrylic polymer resins (e.g., copolymers of acrylic acid and methacrylic acid, methacrylic acid copolymers, methyl methacrylate copolymers, ethoxyethyl methacrylate, cyanoethyl methacrylate, methyl methacrylate copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, methacrylate copolymers, methacrylic acid copolymers, aminoalcohols, etc.). Examples of suitable polyacrylic resins include alkyl methacrylate copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, poly(acrylic acid), poly(methacrylic acid), methacrylic acid alkylamide copolymers, poly(methyl methacrylate), poly(methacrylic acid)(anhydride), methyl methacrylate, polymethacrylate, methyl methacrylate copolymers, poly(methyl methacrylate), poly(methyl methacrylate) copolymers, polyacrylamides, aminoalkyl methacrylate copolymers, poly(methacrylic anhydride), and glycidyl methacrylate copolymers), cellulose acetate phthalate, and polyvinyl acetate phthalate. A specific polyacrylic resin that may be mentioned is polyacrylic resin HB-50.

[0082] Preferably, the pharmaceutical formulation of the second aspect of the invention comprises milled particles of the salt of the invention, the particles having any of the particular particle size distributions described herein, and the formulation further comprises an enteric coating. Thus, in a particular embodiment, in the pharmaceutical formulation of the second aspect of the invention, the salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide is milled so that the particles containing the salt have a particle size distribution defined by a D90 of less than 10 μm, and the formulation further comprises an enteric coating.

[0083] Pharmaceutical formulations that may be mentioned include those in which the salt of the invention is present in a total amount that is at least 1% by weight (or at least 10%, at least 30%, or at least 50% by weight) of the formulation, i.e., the weight ratio of the salt of the invention to the total of the components of the pharmaceutical formulation (i.e., the salt of the invention and all pharmaceutical excipients, such as adjuvants, diluents, and carriers) is at least 1:99 (or at least 10:90, at least 30:70, or at least 50:50).

[0084] As used herein, a "therapeutically effective amount," "effective amount," or "dosage" refers to an amount of a salt of the present invention that is sufficient to produce a desired effect, which may be a therapeutic and / or beneficial effect. The effective amount or dosage will vary depending on the age or general condition of the subject (e.g., a human), the severity of the condition being treated, the particular agent being administered, the duration of treatment, the nature of any concurrent treatments, the pharmaceutically acceptable carrier used, and similar factors within the knowledge and professional judgment of one of ordinary skill in the art. Where appropriate, a "therapeutically effective amount," "effective amount," or "dosage" in any individual case can be determined by one of ordinary skill in the art by reference to relevant texts and literature and / or by conducting routine experimentation. Those of ordinary skill in the art will understand that the therapeutic effect need not be complete or curative, as long as some benefit is provided to the subject.

[0085] Those skilled in the art will understand that the formulations of the present invention can be administered in a variety of dosages (e.g., as one or more of the preparations described above), and suitable dosages can be readily determined by those skilled in the art. The total dosage of the salt of the present invention administered to a subject in need thereof can range from about 0.01 to about 2000 mg / kg body weight per day (mg / kg / day), from about 0.1 to about 500 mg / kg / day, or from about 1 to about 100 mg / kg / day. Such dosages can be, for example, oral dosages of the formulation of the second aspect of the present invention.

[0086] When administered orally, treatment with such a formulation (including capsules containing such a formulation) may involve administration of a unit dose formulation containing about 0.01 mg to about 3,000 mg of a salt of the present invention, for example, about 0.1 mg to about 2,000 mg, or about 1 mg to about 1,000 mg (e.g., about 10 mg to about 500 mg) of the salt of the present invention. Advantageously, treatment may involve administration of the salt of the present invention (including capsules containing the formulation) using a single daily dose. Alternatively, the total daily dose of the salt of the present invention may be administered in divided doses, two, three, or four times daily (e.g., twice daily for the doses described herein, e.g., 100 mg, 250 mg, 500 mg, or 1,000 mg twice daily). A skilled physician will recognize that dosages will vary from subject to subject.

[0087] In a specific embodiment, the daily dose of the salt of the invention administered to a subject ranges from about 1 to about 3000 mg, preferably from about 1 to about 1000 mg.

[0088] The term "about" as used herein when referring to a measurable value, such as the amount of a compound, dosage, time, temperature, etc., refers to a variation of 20%, 10%, 5%, 1%, 0.5%, or no more than 0.1% of the specified amount. In each case, it is contemplated that such a term may be substituted with the notation "±10%" or the like (or by indicating a variation of a particular amount calculated based on the relevant value). It is also contemplated that in each case, such a term may be omitted.

[0089] For the avoidance of doubt, the dose administered to a subject, particularly a human subject, in the context of the present invention should be sufficient to affect a therapeutic response in the subject over a reasonable time frame. Those skilled in the art will recognize that the selection of the exact dose and composition and the most appropriate delivery regimen will also be influenced by, inter alia, the pharmacological properties of the formulation, the nature and severity of the condition being treated, and the physical condition and mental acuity of the recipient, as well as the potency of the particular compound, the age, condition, weight, sex and response of the subject being treated, and the stage / severity of the disease.

[0090] In any event, a physician or other skilled artisan will be able to routinely determine the actual dosage that will be most suitable for an individual subject. The dosages set forth above are exemplary of the average case, and there can, of course, be individual instances where higher or lower dosage ranges are merited, and such are within the scope of this invention.

[0091] Medical Use The salt of the present invention and preparations thereof are also useful as pharmaceuticals.

[0092] The salts of the invention (i.e., salts as defined in the first aspect of the invention, including all embodiments and particular features thereof), and formulations thereof (i.e., formulations as defined in the second aspect of the invention, including all embodiments and particular features thereof) may be particularly useful in the treatment of disorders or conditions ameliorated by activation of AMP-activated protein kinase (AMPK). Accordingly, in a fourth aspect of the invention, there is provided the use of a salt of the invention, or a formulation comprising said salt, as defined herein, in the manufacture of a medicament for the treatment of a disorder or condition ameliorated by activation of AMPK.

[0093] Similarly, there is provided a method of treating a disorder or condition ameliorated by activation of AMPK, comprising administering a therapeutically effective amount of a salt of the invention or a formulation comprising said salt to a subject (e.g., a human) in need thereof. Similarly, there is provided a salt of the invention (or a formulation comprising said salt) for use in a method of treating a disorder or condition ameliorated by activation of AMPK.

[0094] By "activating AMPK," we mean an increase in the steady-state level of phosphorylation of the Thr-172 moiety of the AMPK-α subunit compared to the steady-state level of phosphorylation in the absence of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. Alternatively, or in addition, we mean an increase in the steady-state level of phosphorylation of any other protein downstream of AMPK, such as acetyl-CoA carboxylase (ACC).

[0095] Those skilled in the art will understand that the term "disorders or conditions ameliorated by activation of AMPK" includes cancer, diabetes, cardiovascular disease, hyperinsulinemia and related conditions, conditions / disorders in which fibrosis plays a role, sexual dysfunction, osteoporosis, and neurodegenerative diseases.

[0096] The term "cancer" will be understood by those skilled in the art to include one or more diseases in a class of disorders characterized by the uncontrolled division of cells and their ability to invade other tissues, either by invasion, proliferation, or direct growth into adjacent tissues by implantation at distant sites via metastasis. By "proliferation," we include an increase in the number and / or size of cancer cells. By "metastasis," we mean the movement or migration (e.g., invasiveness) of cancer cells from a primary tumor site within a subject's body to one or more other regions within the subject's body (where the cells may form secondary tumors). 4-Chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can inhibit cancer cell proliferation and cancer cell metastasis.

[0097] Therefore, the formulations of the present invention may be suitable for use in the treatment of any type of cancer, including all tumors (non-solid tumors, preferably solid tumors, such as carcinomas, adenomas, adenocarcinomas, and blood cancers, regardless of organ type). For example, cancer cells may be selected from the group consisting of cancer cells of the breast, bile duct, brain, colon, stomach, reproductive organs, thyroid, hematopoietic system, lung and airway, skin, gallbladder, liver, nasopharynx, nerve cells, kidney, prostate, lymph nodes, and gastrointestinal tract. Preferably, the cancer is selected from the group consisting of colon cancer (including colorectal adenoma), breast cancer (e.g., postmenopausal breast cancer), endometrial cancer, hematopoietic system cancer (e.g., leukemia, lymphoma, etc.), thyroid cancer, kidney cancer, esophageal adenocarcinoma, ovarian cancer, prostate cancer, pancreatic cancer, gallbladder cancer, liver cancer, and cervical cancer. More preferably, the cancer is selected from the group consisting of colon cancer, prostate cancer, and, in particular, breast cancer. When the cancer is a non-solid tumor, it is preferably a hematopoietic tumor such as leukemia (e.g., acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL)). Preferably, the cancer cells are breast cancer cells.

[0098] The term "diabetes" (i.e., diabetes mellitus) is understood by those skilled in the art to refer to both type 1 (insulin-dependent) diabetes and type 2 (non-insulin-dependent) diabetes, both of which are associated with dysfunction of glucose homeostasis. The salts of the invention and formulations thereof may be particularly suitable for use in treating type 1 diabetes and / or type 2 diabetes.

[0099] It will be understood by those skilled in the art that the term "hyperinsulinemia or related conditions" includes hyperinsulinemia, type 2 diabetes, impaired glucose tolerance, insulin resistance, metabolic syndrome, dyslipidemia, childhood hyperinsulinemia, hypercholesterolemia, hypertension, obesity, fatty liver conditions, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, cardiovascular disease, atherosclerosis, cerebrovascular conditions such as stroke, systemic lupus erythematosus, neurodegenerative diseases such as Alzheimer's disease, and polycystic ovary syndrome. Other conditions include progressive renal diseases such as chronic renal failure.

[0100] In particular, the salts of the present invention and formulations thereof may be suitable for use in treating obesity associated with hyperinsulinemia and / or cardiovascular disease associated with hyperinsulinemia.

[0101] The salts of the present invention and formulations thereof may also be suitable for use in treating cardiovascular diseases, such as heart failure, which cardiovascular diseases are not associated with hyperinsulinemia.Similarly, the salts of the present invention and formulations thereof may also be suitable for use in treating obesity that is not associated with hyperinsulinemia.For the avoidance of doubt, the treatment of obesity and / or cardiovascular diseases (such as heart failure) in which AMPK activation may be beneficial is included within the scope of the present invention.

[0102] Conditions / disorders in which fibrosis plays a role include, but are not limited to, scar healing, keloids, scleroderma, pulmonary fibrosis (including idiopathic pulmonary fibrosis), nephrogenic systemic fibrosis, and cardiovascular fibrosis (including endomyocardial fibrosis), systemic sclerosis, liver cirrhosis, ocular macular degeneration, retinal and vitreoretinopathy, Crohn's / inflammatory bowel disease, post-operative scar tissue formation, radiation- and chemotherapy-induced fibrosis, and cardiovascular fibrosis.

[0103] The salts of the invention may also be useful in the treatment of sexual dysfunction (e.g., the treatment of erectile dysfunction). The salts of the invention may also be useful in the treatment of inflammation.

[0104] Neurodegenerative diseases that may be mentioned may include Alzheimer's disease, Parkinson's disease and Huntington's disease, amyotrophic lateral sclerosis, polyglutamine disorders such as spinal-bulbar muscular atrophy (SBMA), dentatorubral-pallidoluysian atrophy (DRPLA), and many spinocerebellar ataxias (SCAs).

[0105] Those skilled in the art will understand that reference to treatment of a particular condition (or, equivalently, treating the condition) takes its ordinary meaning in the medical arts. In particular, the term refers to achieving a reduction in the severity and / or frequency of occurrence of one or more clinical symptoms associated with the condition, as may be determined by a physician treating a subject with or susceptible to such symptoms.

[0106] As used herein, reference to a subject (or subjects) refers to a living subject being treated or receiving a prophylactic medication, including a mammalian (e.g., human) subject. In particular, reference to a subject refers to a human subject.

[0107] For the avoidance of doubt, those skilled in the art will understand that such treatment or prevention is carried out in a subject in need thereof, which may be assessed by one skilled in the art using routine techniques.

[0108] In the context of the present invention, a "subject in need" of a salt of the invention includes a subject suffering from a disorder or condition that is ameliorated by activation of AMPK.

[0109] As used herein, the terms "disease" and "disorder" (and similar terms such as condition, illness, medical problem, etc.) may be used interchangeably.

[0110] Without wishing to be bound by theory, it is believed that the salt forms of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide of the present invention enhance the bioavailability of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide in the systemic circulation. Formulations containing the salts of the present invention have been shown to provide an approximately two-fold increase in the bioavailability of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide under certain circumstances compared to formulations containing 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide in its free base form.

[0111] The salts of the invention (and formulations thereof), whether for use in the above indications or otherwise, may have the advantage that they may be more effective, less toxic, longer acting, more potent, have fewer side effects, be more easily absorbed, and / or have a better pharmacokinetic profile (e.g., higher oral bioavailability and / or lower clearance), and / or possess other useful pharmacological, physical, or chemical properties than other therapies known in the prior art. In particular, the formulations of the invention may have the advantage that they are more effective and / or exhibit advantageous properties in vivo. [Brief explanation of the drawings]

[0112] The following drawings are provided to illustrate various aspects of the inventive concept and are not intended to limit the scope of the invention unless otherwise specified herein.

[0113] [Figure 1] FIG. 1 shows comparative results of an oral pharmacokinetic study using 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide (Compound 1) in suspension and uncoated capsules, and 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide sodium salt (Compound 2) and 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide potassium salt (Compound 3) separately in uncoated capsules. [Figure 2] 1 shows comparative results of an oral pharmacokinetic study using Compound 1 in suspension and in enteric-coated capsules, and Compounds 2 and 3 separately in enteric-coated capsules. [Figure 3] Absolute and relative Cmax results are shown for formulations of Compound 1 as a suspension and in uncoated capsules, and Compound 2 and Compound 3 separately in uncoated capsules. [Figure 4] Same as above. [Figure 5] Absolute and relative AUC results are shown for formulations of Compound 1 as a suspension and in uncoated capsules, and Compound 2 and Compound 3 separately in uncoated capsules. [Figure 6] Same as above. [Figure 7] Absolute and relative Cmax results are shown for formulations of Compound 1 as a suspension and in enteric coated capsules, and Compound 2 and Compound 3 separately in enteric coated capsules. [Figure 8] Same as above. [Figure 9] Absolute and relative AUC results are shown for formulations of Compound 1 as a suspension and in enteric coated capsules, and Compound 2 and Compound 3 separately in enteric coated capsules. [Figure 10] Same as above. [Example]

[0114] Abbreviation AUC 0-t : Area under the concentration-time curve from time zero to the last quantifiable concentration AUC 0-∞ : Area under the concentration-time curve from time zero to infinity bw: weight CE: Collision energy CL: Clearance C max :Peak plasma concentration. CXP: Collision exit potential DLS: Dynamic Light Scattering DP: Declustering potential EP: Entrance potential h: time HPLC: High-performance liquid chromatography ISD: Internal standard K2EDTA Dipotassium ethylenediaminetetraacetic acid LC: liquid chromatography LC-MS / MS: Liquid chromatography-(tandem) mass spectrometry LS: Light scattering MRT: Mean residence time. min:minutes PVP K30: Polyvinylpyrrolidone K30 rpm: revolutions per minute RT: room temperature T 1 / 2 :Half-life T max : Time to reach peak plasma concentration v / v: volume / volume w / v: weight / volume

[0115] The present invention is further illustrated by reference to the following examples, which are not intended to limit the scope of the invention.

[0116] material 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide (Compound 1), 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide sodium salt (Compound 2), and 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide potassium salt (Compound 3) were prepared by Anthem Biosciences.

[0117] Sodium-docusate, PVP K30, and mannitol were supplied by Sigma-Aldrich.

[0118] Preparation of 1-4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide sodium salt (compound 2)

[0119] Scheme:

[0120] [ka]

[0121] procedure:

[0122] To a suspension of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide (100 g, 0.2629 mol) in isopropanol (1.0 L) was slowly added a solution of sodium hydroxide (11.56 g, 0.2891 mol) in water (100 mL) at 25±5°C. The mass was stirred at 25±5°C for 3 hours and cooled to 5±5°C. The mass was stirred at 5±5°C for 3 hours and filtered to collect the solid. The solid was washed with isopropanol (300 mL) and dried under reduced pressure at 35±5°C for 8 hours. The dried solid was dried at 4.0 kg / cm 2 Primary pressure: 7.0 kg / cm 2 After micronization twice using an air jet mill equipped with a secondary pressure of 1000 kJ and a screw feeder at 8 RPM, the desired sodium salt was isolated as a white solid (50 g, 48%).

[0123] Preparation of 2-4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide potassium salt (compound 3)

[0124] Scheme:

[0125] [ka]

[0126] procedure:

[0127] To a suspension of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide (100 g, 0.2629 mol) in isopropanol (1.0 L) was slowly added a solution of potassium hydroxide (16.22 g, 0.2891 mol) in water (100 mL) at 25±5°C. The mass was stirred at 25±5°C for 3 hours and cooled to 5±5°C. The mass was stirred at 5±5°C for 3 hours and filtered to collect the solid. The solid was washed with isopropanol (300 mL) and dried under reduced pressure at 35±5°C for 8 hours. The dried solid was pressurized at 4.0 kg / cm 2 Primary pressure: 7.0 kg / cm 2 After micronization twice using an air jet mill equipped with a secondary pressure of 1000 kJ and a screw feeder at 8 RPM, the desired potassium salt was isolated as a white solid (55 g, 50%).

[0128] Example 1 - Ground product

[0129] Milling was carried out for Examples 1a to 1c using an air jet mill (equipment code CP-AJM-01; Promas engineers) with the following parameters:

[0130] [Table 1]

[0131] Micronization was repeated for all examples by another air jet milling run using the same parameters, which allowed for particle size reduction to a D90 of less than 10 μm.

[0132] Particle size analysis

[0133] [Table 2]

[0134] [Table 3]

[0135] [Table 4]

[0136] preparation

[0137] Four drops of Tween 20 were added to 25 mL of water and the mixture was sonicated for 3 minutes to form a dispersant solution.

[0138] 0.05 g of the ground material was transferred to a 250 mL glass beaker. 25 mL of dispersant solution (Tween 20 / water mixture) was added to the beaker and rotated continuously for 2-3 minutes. The suspension was transferred to a measuring unit, and particle size distribution measurements were performed in triplicate.

[0139] [Table 5]

[0140] Examples 2-8 - Single Oral Pharmacokinetic Study in Rabbits

[0141] The studies detailed below (and referred to as Examples 2-8) were conducted in male New Zealand White rabbits to provide comparative single-dose oral pharmacokinetic data for Compound 1, and the sodium and potassium salts of that compound, using uncoated and enteric-coated capsules.

[0142] Preparation of formulations

[0143] 1. Capsules

[0144] Ready-to-use enteric coated and uncoated capsules were used and were obtained from CapsulCN International Co., Ltd.

[0145] Compound 1 was prepared using the process described in WO2011 / 004162 and milled as described in Example 1c. Uncoated or enteric-coated gelatin capsules were individually filled with 180 mg of dry-milled Compound 1, along with accompanying excipients, as indicated. Both capsule types were filled with Compound 1 along with 1.8 mg sodium docusate, 0.18 mg PVP K30, and 9 mg mannitol.

[0146] Similarly, for Compound 2, uncoated or enteric-coated gelatin capsules were individually filled with 180 mg of dry-milled Compound 2 (obtained as in Example 1f) along with 1.8 mg of sodium doxorubicin, 0.18 mg of PVP K30, and 9 mg of mannitol.

[0147] For Compound 3, uncoated or enteric coated gelatin capsules were individually filled with 90 mg of dry-milled Compound 3 (obtained in Example 1i) along with 0.90 mg sodium docusate, 0.09 mg PVP K30, and 4.50 mg mannitol.

[0148] Once prepared, the capsules were stored in a desiccator at 19-25°C before administration to the animals.

[0149] Formulation details are summarized in Table 2.

[0150] 2. Suspension

[0151] Compound 1 was made using the process described in WO2011 / 004162.

[0152] A 2% w / v methylcellulose solution in 4 mM phosphate buffer pH 7.4 was prepared.

[0153] 40 mL of 2% w / v methylcellulose solution was added to a 250 mL Erlenmeyer flask along with 10 g of 2 mm glass beads and vigorously stirred. Compound 1 (720 mg) was slowly added to the solution, and the mixture was continuously stirred for 1 hour. The homogenate was transferred to another flask, and its pH was recorded. A suspension formulation was prepared prior to administration to animals.

[0154] Animal Management

[0155] Rabbits (New Zealand White, male) were housed under standard laboratory conditions in an environmentally monitored, climate-controlled room with adequate fresh air supply (10-15 air changes per hour), room temperature (22 ± 3°C), relative humidity (30-70%), and a 12-h light and 12-h dark cycle. Temperature and relative humidity were recorded daily.

[0156] Each animal was housed in a standard stainless steel rabbit cage SS-304 (size: L24" x B18" x H18") equipped with a stainless steel mesh and a removable bottom tray for waste disposal, a food hopper for holding pelleted food, a holder for a drinking water bottle and siphon tube, and a label holder. Clean, sterilized corncobs were provided as bedding material.

[0157] Animals were fed Krishna Valley Agrotech rabbit chow ad libitum throughout the acclimatization and experimental period.

[0158] Water was provided ad libitum throughout the acclimation and experimental periods. Water from an Aquaguard water filter and purifier was autoclaved and provided in polypropylene water bottles with stainless steel sipper tubes.

[0159] Acclimation

[0160] Animals were allowed to acclimate to the facility room conditions for a minimum of 1 week (7 days) and were observed daily for clinical signs. Veterinary examinations of all animals were performed on the day of receipt, daily, and on the day of randomization.

[0161] Grouping

[0162] Animal grouping was performed by weight stratification and randomization. The animals selected for the study were weighed and grouped according to their weight range. These weight-stratified rabbits were distributed equally among all study groups, if possible, so that the weight variation of the animals used did not exceed ±20% of the mean weight. Grouping was performed one day before the start of treatment.

[0163] [Table 6]

[0164] dose control

[0165] Healthy adult male New Zealand White rabbits, approximately 2-3 months old, were used in the experiments after 7 days of acclimation.

[0166] 1. Capsules

[0167] A soft plastic dosing tube was used to administer the filled capsules. The filled capsules were inserted into the dosing tube so that the short end of the capsule protruded slightly beyond the tip of the tube. The tip of the capsule was dipped in mineral oil to aid in swallowing.

[0168] The head was grasped firmly by holding down the upper jaw with one hand. The dosing tube containing the capsule was inserted behind the incisors. The dosing tube was slid straight to the back of the mouth. The capsule was expelled by pressing the plunger on the dosing tube. The dosing tube was removed and the rabbit's mouth was closed. The neck was gently stroked to facilitate swallowing.

[0169] 2. Suspension

[0170] An infant feeding tube was used to administer the suspension. The feeding tube was inserted through the rabbit's mouth into the esophagus and stomach, and it was confirmed that it was not inserted into the trachea before administration. The suspension of Compound 1 was administered through the feeding tube. After administration of the suspension, approximately 2.0–2.5 mL of drinking water was administered to flush out the contents of the feeding tube.

[0171] Blood sampling

[0172] Animals were restrained in a rabbit restrainer, and blood samples (400–500 μL / time point) were collected via the middle ear artery at 0.16, 0.25, 0.50, 1.0, 2.0, 4.0, 6.0, 8.0, 24.0, 48.0, and 72 hours post-dose. The collected blood specimens were centrifuged at 4000 or 6000 rpm at 4°C for 10 minutes, and plasma samples were separated and stored at -80°C until analysis.

[0173] bioanalysis

[0174] Analyte concentrations of Compound 1 in New Zealand White rabbits were determined using an API 3200 Q-trap LC-MS / MS system.

[0175] Method overview

[0176] Chromatographic separation was performed on a Zorbax C18, 50 × 4.6 mm, 5 μm column using a gradient elution of methanol-0.1% formic acid as the mobile phase. The flow rate was 1.0 mL min -1 The calibration curve was set to 0. Detection was achieved by a triple quadrupole tandem mass spectrometer with multiple reaction monitoring (MRM) scanning via an electrospray ionization source applied in positive mode. The mass transition ion pairs optimized for quantitation were m / z 379.999 → 125.000 for compound 1 and m / z 376.165 → 165.00 for ISD (haloperidol). The calibration plot was linear over the range of 11.062 to 20594.820 ng / mL.

[0177] Buffer solution (0.1% formic acid)

[0178] Approximately 1.0 mL of formic acid was added to 999 mL of ultrapure water Type 1 to prepare a buffer solution. This solution was stored at room temperature and used within two days of preparation.

[0179] Dilution solvent (methanol:water, 80:20% v / v)

[0180] 800 mL of methanol and 200 mL of ultrapure water type 1 were added to the reagent bottle, mixed thoroughly, and sonicated. The solution was stored at room temperature and used within 7 days of preparation.

[0181] Rinse solvent (methanol:water, 50:50% v / v)

[0182] 500 mL of methanol and 500 mL of ultrapure water type 1 were added to the reagent bottle, mixed thoroughly, and sonicated. The solution was stored at room temperature and used within 3 days of preparation.

[0183] Preparation of Compound 1 stock solution

[0184] 3.044 mg of Compound 1 was weighed out, transferred to a 5.0 mL volumetric flask, and dissolved in 5 mL of methanol to obtain a stock solution of 605.756 μg / mL. The final concentration of Compound 1 was corrected according to the potency of the standard and the amount actually weighed.

[0185] Internal Standard Stock Solution

[0186] Approximately 2.00 mg of haloperidol (Sigma-Aldrich) was weighed into a 5.0 mL volumetric flask and dissolved in methanol to obtain a 400 μg / mL internal standard stock solution. The final concentration of haloperidol was corrected according to the potency of the standard.

[0187] Preparation of internal standard (haloperidol) working solution.

[0188] Approximately 0.250 mL of the internal standard stock solution was diluted to 10 mL using dilution solvent (methanol / water, 80:20) to give a solution of approximately 10 μg / mL.

[0189] Preparation of a calibration curve for compound 1 in plasma

[0190] Calibration curve standards were prepared in the range of 11.062 to 20594.820 ng / mL in plasma (prepared concentrations: 11.062, 20.112, 40.224, 80.449, 160.897, 321.794, 643.588, 1287.176, 2574.353, 5148.705, 10297.410, and 20594.820 ng / mL) by spiking aqueous analyte standards into blank plasma.

[0191] Quality control samples LQC, MQC, and HQC within the calibration curve were prepared for Compound 1 in plasma (prepared concentrations: 40.224, 5148.705, and 10297.410 ng / mL) by spiking blank plasma with the appropriate aqueous analyte standards.

[0192] Liquid-liquid extraction of plasma samples

[0193] 10 μL of ISD working solution (approximately 10 μg / mL) was added to the RIA vial. Exactly 50 μL of rabbit plasma was added from a polypropylene capped tube / vial to the RIA vial and vortexed for 30-40 seconds. · 2.5 mL of TBME was added to the RIA vial, which was then vortexed for 10 minutes at 2000 rpm using a vibramax shaker. The samples were centrifuged at 4000 rpm for 10 minutes at 4°C. 2 mL of the organic layer was separated and evaporated to dryness using a turbo evaporator at 50°C for 20 minutes. The sample residue was reconstituted with 200 μL of reconstitution solvent (methanol). The reconstituted sample was then transferred to an autosampler vial. · 10 μL of the reconstituted sample was injected into an API 3200 Q Trap LC-MS / MS system.

[0194] [Table 7]

[0195] Data analysis

[0196] The plasma concentration data for each time point for the analyte Compound 1 were used for pharmacokinetic analysis, which was performed using the non-compartmental analysis (NCA) module of Phoenix WinNonlin 6.3 software, and the following pharmacokinetic parameters were determined:

[0197] C max T max AUC 0-t AUC 0-∞ T 1 / 2: MRT

[0198] Chromatograms of data acquired using Analyst software version 1.6.1 were processed by the peak area ratio method. A weighting factor of 1 / ×2 was used. The concentration of unknown substances was calculated using the following formula:

[0199] y=mx+c where x = drug concentration; m = slope of the calibration curve; y = peak area ratio; and c = intercept of the calibration curve.

[0200] result

[0201] The results for the single oral pharmacokinetic study in rabbits are tabulated below in Tables 3-8 and graphically depicted in Figures 1-10.

[0202] The results show that there is a 50-100% increase in systemic exposure of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide when it is administered as the dry-milled sodium salt (Compound 2) compared to the free base active ingredient (Compound 1) (compare Examples 2 and 3, and also Examples 5 and 6, Figures 3-10). The dry-milled potassium salt (Compound 3) showed a more modest improvement compared to the milled free base active ingredient (compare Examples 2 and 4, and also Examples 5 and 7, Figures 3-10).

[0203] When compared with a standard suspension of the active ingredient (Example 8), there was a surprisingly large (up to 6-fold) increase in the systemic exposure of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide when the active ingredient was administered as the dry-milled sodium salt (Compound 2) in either uncoated or enteric-coated capsules.

[0204] Thus, the systemic exposure of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide is increased by administration of the compound in the form of its alkali metal salt. The greatest improvement in systemic exposure was observed when the drug was administered in the form of its sodium salt.

[0205] [Table 8]

[0206] [Table 9]

[0207] [Table 10]

[0208] [Table 11]

[0209] [Table 12]

[0210] [Table 13]

[0211] Examples 9 and 10

[0212] The following studies (designated Examples 9 and 10) provide comparative single-dose oral pharmacokinetic data for Compound 2 (the sodium salt of Compound 1) using enteric-coated capsules in male New Zealand White rabbits in the presence of a pH modifier (magnesium oxide). The studies were conducted using the methods described above for Examples 2-8, except where indicated below.

[0213] Enteric coated gelatin capsules were individually filled with: Example 9: Compound 2 (62.5 mg), sodium docusate (6.25 mg), PVP K30 (0.625 mg) and magnesium oxide (125 mg). Example 10: Compound 2 (62.5 mg), sodium docusate (6.25 mg), PVP K30 (0.625 mg) and mannitol (125 mg).

[0214] Animals were given a single dose of Compound 2 at 25 mg / kg bw.

[0215] result

[0216] The results for the single-dose oral pharmacokinetic study in rabbits are tabulated below in Table 9.

[0217] The results show that there is an increase in the systemic exposure of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide when Compound 2 is formulated with an alkaline excipient compared to formulation in the absence of the alkaline excipient.

[0218] [Table 14]

[0219] Example 11

[0220] The following study was a single-dose oral pharmacokinetic study of Compound 2 using enteric-coated capsules in male New Zealand White rabbits in the presence of a pH adjuster (magnesium oxide). The study was conducted using the methods described above for Examples 2-8, except where indicated below.

[0221] Enteric coated gelatin capsules were individually filled with Compound 2 (106 mg, obtained in Example 1f), sodium docusate (10.6 mg), PVP K30 (1.1 mg), and magnesium oxide (100 mg).

[0222] Animals received a single dose of 53 mg / kg bw of Compound 2 (equivalent to 50 mg / kg of Compound 1).

[0223] result

[0224] The results for the single-dose oral pharmacokinetic study in rabbits are tabulated below in Table 10.

[0225] The results show that when compound 2 is formulated with alkaline excipients, the systemic exposure of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide is significant.

[0226] [Table 15]

[0227] Example 12

[0228] The following illustrates a representative pharmaceutical tablet dosage form containing Compound 2 for therapeutic or prophylactic use in humans.

[0229] [Table 16]

[0230] These formulations may be obtained by conventional procedures well known in the pharmaceutical art. The tablets may also be enteric coated by conventional means, for example to provide a coating of polymethacrylate.

Claims

1. 1. A composition for oral administration comprising the sodium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide in a daily dose of 10 mg to 1000 mg, for treating a disorder or condition ameliorated by activation of AMP-activated protein kinase (AMPK) in a human in need thereof.

2. the daily dose of the sodium salt is 10 mg to 500 mg; The composition of claim 1.

3. the daily dose of the sodium salt is 200 mg to 1000 mg; The composition of claim 1.

4. the daily dose of the sodium salt is 200 mg to 500 mg; The composition of claim 1.

5. the daily dose of the sodium salt is 500 mg to 1000 mg; The composition of claim 1.

6. The disorder or condition is cancer, diabetes, cardiovascular disease, hyperinsulinemia and related conditions, conditions or disorders in which fibrosis plays a role, sexual dysfunction, osteoporosis, or neurodegenerative diseases; The composition according to any one of claims 1 to 5.

7. The disorder or condition is hyperinsulinemia, type 2 diabetes, impaired glucose tolerance, insulin resistance, metabolic syndrome, dyslipidemia, childhood hyperinsulinemia, hypercholesterolemia, hypertension, obesity, fatty liver conditions, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, cardiovascular disease, atherosclerosis, cerebrovascular conditions, stroke, systemic lupus erythematosus, neurodegenerative diseases, Alzheimer's disease, or polycystic ovary syndrome; The composition according to any one of claims 1 to 5.

8. The sodium salt is administered in a tablet or capsule. The composition according to any one of claims 1 to 5.

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