Salts of heterocyclic inhibitors of monocarboxylate transporter 4 for treating diseases

Abstract

Disclosed herein are novel salts of substituted pyrazole compounds that inhibit the activity of the monocarboxylate transporter MCT4 or its mutants. Also disclosed are compounds, pharmaceutical compositions, and methods of treatment for MCT4-mediated diseases, such as inflammatory and proliferative disorders.

Description

[Technical Field]

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 370,972, filed August 10, 2022, the contents of which are incorporated by reference as if set forth in their entirety herein. [Background technology]

[0002] Lactate export from glycolytic cells is normally mediated by the monocarboxylate transporter MCT4, which has a weak affinity for lactate (K m = 28 mM), which, in combination with a high turnover rate, allows for the rapid export of large amounts of lactate. MCT4 expression is typically restricted to highly glycolytic tissues such as white muscle fibers, lymphocytes, astrocytes, and Sertoli cells. While MCT4 is absent in most normal tissues, in many cancer indications, including colorectal cancer, glioma, head and neck cancer, triple-negative breast cancer, prostate cancer, KRAS-mutated lung cancer, liver cancer, and kidney cancer, MCT4 expression is highly upregulated and correlates with reduced survival.

[0003] The correlation between MCT4 expression and poor cancer outcomes appears to be related to important functional consequences in multiple cancer models. Stable expression of MCT4 is highly tumorigenic in respiratory-impaired, Ras-transformed fibroblast xenograft models. Conversely, silencing MCT4 slows or eliminates tumor growth in xenograft models of breast cancer, colon cancer, and glioma. In xenograft models of breast cancer and colon cancer, MCT4 expression is required for inflammatory cytokine IL-8-mediated angiogenesis. MCT4 has also been shown to play an important role in cancer cell migration, invasion, and various aspects of the Warburg effect (e.g., growth on glucose, extracellular acidification, and lactate secretion).

[0004] Inhibition of MCT4-mediated lactate export may be an effective strategy to reduce the Warburg effect in cancer. Unfortunately, no potent and selective MCT4 inhibitors have been reported. Moderate to weak MCT4 inhibitors are known (e.g., phloretin and α-CN-4-OH-cinnamate); however, these compounds indiscriminately inhibit several other transporters, including MCT1.

[0005] Novel salts and pharmaceutical compositions have been discovered that are found to inhibit MCT4, along with methods of synthesizing and using the salts, including methods of treating MCT4-mediated diseases in patients by administering the compounds. Summary of the Invention [Means for solving the problem]

[0006] Structural formula I [ka] Form A of the compound is provided.

[0007] In addition, the amorphous structural formula I [ka] Also provided is a compound of the formula:

[0008] Furthermore, structural formula II [ka] Also provided is a compound of the formula:

[0009] Also, structural formula II [ka] Also provided is Form A of the compound:

[0010] Furthermore, structural formula II [ka] Also provided is Form B of the compound:

[0011] Also provided are pharmaceutical compositions comprising a compound described herein and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0012] Also provided is a method of inhibiting the activity of the monocarboxylate transporter MCT4, or a variant thereof, in a biological sample, comprising contacting the biological sample with a compound described herein.

[0013] Also provided is a method of inhibiting the activity of the monocarboxylate transporter MCT4, or a variant thereof, in a patient, comprising administering to the patient a compound described herein.

[0014] Also provided is a method for selectively inhibiting the activity of the monocarboxylate transporter MCT4 or a variant thereof over the monocarboxylate transporter MCT1 or a variant thereof in a patient, comprising administering to the patient a compound described herein.

[0015] Also provided is a method for treating a monocarboxylate transporter MCT4-mediated disorder in a subject in need thereof, comprising administering to the patient a compound described herein.

[0016] Also provided is a method for treating a monocarboxylate transporter MCT4-mediated disorder in a subject in need thereof, comprising sequentially or co-administering a compound described herein with another therapeutic agent.

[0017] Also provided are methods for achieving an effect in a patient, comprising administering to the patient a therapeutically effective amount of a compound described herein, wherein the effect is selected from the group consisting of reduced triglycerides, reduced cholesterol, and reduced hemoglobin A1c.

[0018] These and other aspects of the present invention will become apparent upon reference to the following description, and to this end, various references are set forth herein which describe in more detail certain background information, procedures, compounds, and / or compositions, each of which is incorporated herein by reference in its entirety. [Brief explanation of the drawings]

[0019] [Figure 1] 1 shows the FT-Raman spectrum of the compound of formula I. [Figure 2] 1 shows DSC and TGA traces of the compound of formula I. [Figure 3] 1 shows the XRPD diffractogram of the compound of formula I. [Figure 4] 1 shows the 1H-NMR spectrum of the compound of structural formula I. [Figure 5] 1 shows the FT-Raman spectrum of Form A of the compound of structural formula II. [Figure 6] 1 shows DSC and TGA traces of Form A of the compound of structural formula II. [Figure 7] 1 shows an XRPD diffractogram of Form A of the compound of structural formula II. [Figure 8] 1 shows the 1H-NMR spectrum of Form A of the compound of structural formula II. [Figure 9] 1 shows the dynamic vapor sorption plot of Form B of the compound of structural formula II. [Figure 10] 1 shows DSC and TGA traces of Form B of the compound of structural formula II. [Figure 11] 1 shows an XRPD diffractogram of Form B of the compound of structural formula II. [Figure 12] 1 shows the 1H-NMR spectrum of Form B of the compound of structural formula II. [Figure 13] 1 shows the mean plasma concentrations of the compound of formula I following oral administration of 20, 60, and 200 mg / kg to male rats. [Figure 14] 1 shows the mean plasma concentrations of the compound of formula I following oral administration of 20, 60, and 200 mg / kg to female rats. [Figure 15]1 shows the mean plasma concentrations of the compound of formula II following oral administration of 25, 75, and 250 mg / kg to male rats. [Figure 16] 1 shows the mean plasma concentrations of the compound of formula II following oral administration of 25, 75, and 250 mg / kg to female rats. [Figure 17] 1 shows the mean plasma concentrations of the compound of formula I following oral administration at 3 mg / kg to male dogs. [Figure 18] 1 shows the mean plasma concentrations of the compound of formula II following oral administration at 3 mg / kg to male dogs. [Figure 19] 1 shows plasma concentrations of the compound of formula II following oral administration at 3 mg / kg to male dogs. [Figure 20] 1 shows plasma concentrations of the compound of formula II following oral administration at 30 mg / kg to male dogs. [Figure 21A] 1 shows a schematic diagram of potential dose groups for Parts A and C of the Phase 1 protocol. [Figure 21B] A schematic diagram of potential dose groups for Parts A and C of the Phase 1 protocol is shown (continuation of Figure 21A). [Figure 22] 1 shows a schematic diagram of potential dose groups for Parts B and D of the Phase 1 protocol. [Figure 23]

[0023] Figure 1 shows the results of a bleomycin-induced pulmonary fibrosis model in mice. Compound 1 (VB253) was administered at 3 mg / kg BID; pirfenidone was administered at 100 mg / kg BID; and nintedanib was administered at 50 mg / kg QD, all by oral gavage. DETAILED DESCRIPTION OF THE INVENTION

[0020] Structural formula I [ka] Form A of the compound is provided.

[0021] The compound of structural formula I is also referred to herein as Compound 1 or 2-((1-(2-(azetidin-1-yl)phenyl)-5-(3-cyclobutoxyphenyl)-1H-pyrazol-3-yl)methoxy)-2-methylpropanoic acid. In certain embodiments, Compound 1 is administered as a pharmaceutically acceptable salt, such as the tris salt, of 2-((1-(2-(azetidin-1-yl)phenyl)-5-(3-cyclobutoxyphenyl)-1H-pyrazol-3-yl)methoxy)-2-methylpropanoic acid.

[0022] In certain embodiments, Form A of the compound of structural formula I is unsolvated.

[0023] In certain embodiments, Form A of the compound of structural formula I has differential scanning calorimetry data that exhibits a first-order melting endotherm with an onset at about 157°C.

[0024] In certain embodiments, Form A of the compound of structural formula I has a differential scanning calorimetry trace substantially as shown in FIG.

[0025] In certain embodiments, Form A of the compound of structural formula I has a TGA trace substantially as shown in FIG.

[0026] In certain embodiments, Form A of the compound of structural formula I has an X-ray powder diffraction (XRPD) pattern substantially as shown in FIG.

[0027] In certain embodiments, Form A of the compound of structural formula I has an FT-Raman spectrum substantially as shown in FIG.

[0028] Also provided is Form A of the compound of structural formula I prepared by the processes described herein.

[0029] In addition, the amorphous structural formula I [ka] Also provided is a compound of the formula:

[0030] Furthermore, structural formula II [ka] Also provided is a compound of the formula:

[0031] The compound of structural formula II is also referred to herein as the Tris salt or Compound 1 Tris salt.

[0032] In certain embodiments, the compound of structural formula II has a 1:1 active ingredient:counterion stoichiometry.

[0033] Also, structural formula II [ka] Also provided is Form A of the compound:

[0034] In certain embodiments, Form A of the compound of structural formula II is unsolvated.

[0035] In certain embodiments, Form A of the compound of structural formula II has a 1:1 active ingredient:counterion stoichiometry.

[0036] In certain embodiments, Form A of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern with peaks at about 7.79, 15.61, 16.71, 20.00, and 20.88±0.3 degrees 2θ, where the XRPD is measured using an incident beam of Cu radiation.

[0037] In certain embodiments, Form A of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern with peaks at about 7.79, 12.59, 15.61, 16.71, 20.00, 20.88, and 21.50±0.3 degrees 2θ, where the XRPD is measured using an incident beam of Cu radiation.

[0038] In certain embodiments, Form A of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern with peaks at about 7.79, 12.18, 12.59, 15.61, 16.71, 17.38, 17.72, 19.16, 20.00, 20.88, and 21.50±0.3 degrees 2θ, where the XRPD is measured using an incident beam of Cu radiation.

[0039] In certain embodiments, Form A of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern with peaks at d-spacings of about 11.34, 5.67, 5.30, 4.44, and 4.25±0.3 Å, where the XRPD is measured using an incident beam of Cu radiation.

[0040] In certain embodiments, Form A of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern with peaks at d-spacings of about 11.34, 7.02, 5.67, 5.30, 4.44, 4.25, and 4.13±0.3 Å, where the XRPD is measured using an incident beam of Cu radiation.

[0041] In certain embodiments, Form A of the compound of Structural Formula II has an X-ray powder diffraction (XRPD) pattern with peaks at d-spacings of about 11.34, 7.26, 7.02, 5.67, 5.30, 5.10, 5.00, 4.63, 4.44, 4.25, and 4.13 Å, where the XRPD is measured using an incident beam of Cu radiation.

[0042] In certain embodiments, Form A of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern substantially as shown in FIG.

[0043] In certain embodiments, Form A of the compound of structural formula II has peaks at about 1604, 1438, and 995 cm -1 It is characterized by the presence of an FT-Raman peak of

[0044] In certain embodiments, Form A of the compound of structural formula II has peaks at about 1604, 1438, 1372, 995, 332, 234, and 173 cm-1 It is characterized by the presence of an FT-Raman peak of

[0045] In certain embodiments, Form A of the compound of structural formula II has an FT-Raman spectrum substantially as shown in FIG.

[0046] In some embodiments, the compound is characterized by a weight loss of 0.5% or less between about 25°C and 150°C by thermogravimetric analysis (TGA).

[0047] In some embodiments, Form A of the compound of structural formula II has a TGA trace substantially as shown in FIG.

[0048] In certain embodiments, Form A of the compound of structural formula II has differential scanning calorimetry data that exhibits a melting endotherm with an onset at about 148°C.

[0049] In certain embodiments, Form A of the compound of structural formula II has a differential scanning calorimetry trace substantially as shown in FIG.

[0050] Also provided is a process for preparing Form A of the compound of structural formula II, comprising combining a compound of structural formula I with tris(hydroxymethyl)aminomethane in a solvent and isolating Form A of the compound of structural formula II. In some embodiments, the solvent is selected from water, acetone, and acetonitrile, or a mixture thereof. In some embodiments, the combining step is performed at room temperature.

[0051] Also provided is Form A of the compound of structural formula II prepared by the processes described herein.

[0052] Also, structural formula II [ka] Also provided is Form B of the compound:

[0053] In certain embodiments, Form B of the compound of structural formula II is unsolvated.

[0054] In certain embodiments, Form B of the compound of structural formula II has a 1:1 active ingredient:counterion stoichiometry.

[0055] In certain embodiments, Form B of the compound of structural formula II has a differential scanning calorimetry trace substantially as shown in FIG.

[0056] In certain embodiments, Form B of the compound of structural formula II has a dynamic vapor sorption plot substantially as shown in FIG.

[0057] In certain embodiments, Form B of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern with peaks at about 9.29, 9.70, 16.36, 19.12, and 20.15 degrees 2θ, where the XRPD is measured using an incident beam of Cu radiation.

[0058] In certain embodiments, Form B of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern with peaks at about 9.29, 9.70, 10.03, 16.36, 19.12, 19.49, 19.61, 20.15, and 21.68 degrees 2θ, where the XRPD is measured using an incident beam of Cu radiation.

[0059] In certain embodiments, Form B of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern with peaks at about 9.29, 9.70, 10.03, 11.14, 11.73, 16.36, 16.71, 19.12, 19.49, 19.61, 20.15, 20.52, 20.73, and 21.68 degrees 2θ, where the XRPD is measured using an incident beam of Cu radiation.

[0060] In certain embodiments, Form B of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern with peaks at d-spacings of about 9.51, 9.11, 5.41, 4.64, and 4.40 Å, where the XRPD is measured using an incident beam of Cu radiation.

[0061] In certain embodiments, Form B of the compound of Structural Formula II has an X-ray powder diffraction (XRPD) pattern with peaks at d-spacings of about 9.51, 9.11, 8.81, 5.41, 4.64, 4.55, 4.52, 4.40, and 4.10 Å, where the XRPD is measured using an incident beam of Cu radiation.

[0062] In certain embodiments, Form B of the compound of Structural Formula II has an X-ray powder diffraction (XRPD) pattern with peaks at d-spacings of about 9.51, 9.11, 8.81, 7.93, 7.54, 5.41, 5.30, 4.64, 4.55, 4.52, 4.40, 4.32, 4.28, and 4.10 Å, where the XRPD is measured using an incident beam of Cu radiation.

[0063] In certain embodiments, Form B of the compound of structural formula II has an X-ray powder diffraction (XRPD) pattern substantially as shown in FIG.

[0064] In certain embodiments, Form B of the compound of structural formula II is characterized by a monoclinic lattice and a P21 / c space group with three axial unit cell lengths of about (a) 26.526 A, (b) 5.940 A, and (c) 19.055 A, and three unit cell angles of about (a) 90.00°, (β) 90.00°, and (γ) 93.123°.

[0065] In certain embodiments, Form B of the compound of structural formula II has peaks at about 1601, 1545, 1468, 1437, 999, 995, and 234 cm -1 It is characterized by the presence of an FT-Raman peak of

[0066] In certain embodiments, Form B of the compound of structural formula II has peaks of about 2946, 1601, 1545, 1507, 1468, 1437, 1374, 1345, 1043, 999, 995, 284, 234, and 186 cm -1 It is characterized by the presence of an FT-Raman peak of

[0067] In certain embodiments, Form B of the compound of structural formula II has differential scanning calorimetry data that exhibits a melting endotherm with an onset at about 148°C.

[0068] In some embodiments, the compound is characterized by a weight loss of 0.1% or less between about 25°C and 145°C by thermogravimetric analysis (TGA).

[0069] In some embodiments, Form B of the compound of structural formula II has a TGA trace substantially as shown in FIG.

[0070] Also provided is a process for making Form B of the compound of structural formula II, which comprises stirring Form A of the compound of structural formula II with a suitable solvent, followed by adding seed crystals of Form B of the compound of structural formula II, and isolating Form B of the compound of structural formula II. Also provided is Form B of the compound of structural formula II prepared by the process described herein.

[0071] Also provided are embodiments in which any of the above-described embodiments may be combined with any one or more of these embodiments, provided that such combinations are not mutually exclusive.

[0072] As used herein, two embodiments are "mutually exclusive" if one is defined as different from the other. For example, an embodiment in which two groups are linked to form a cycloalkyl is mutually exclusive from an embodiment in which one group is ethyl and the other group is hydrogen. Similarly, an embodiment in which one group is CH2 is mutually exclusive from an embodiment in which the same group is NH.

[0073] Certain compounds disclosed herein have useful MCT4 inhibitory activity and can be used to treat or prevent diseases or conditions in which MCT4 plays an active role. Accordingly, in broad aspects, certain embodiments also provide pharmaceutical compositions comprising one or more compounds disclosed herein together with a pharmaceutically acceptable carrier, as well as methods for making and using these compounds and compositions. Certain embodiments provide methods for inhibiting MCT4. Other embodiments provide methods for treating an MCT4-mediated disorder in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a compound or composition according to the invention. Also provided is the use of certain compounds disclosed herein for use in the manufacture of a medicament for treating a disease or condition ameliorated by MCT4 inhibition.

[0074] Methods for inhibiting at least one MCT4 function are provided, comprising contacting MCT4 with a compound described herein. Cell phenotype, cell proliferation, MCT4 activity, changes in biochemical outputs caused by active MCT4, MCT4 expression, or binding of MCT4 to a natural binding partner can be monitored. Such methods may be disease treatment modes, biological assays, cellular assays, biochemical assays, etc.

[0075] Also provided herein is a method for treating an MCT4-mediated disease, comprising administering to a patient in need thereof a therapeutically effective amount of a compound disclosed herein.

[0076] In certain embodiments, the disease is selected from proliferative inflammatory diseases.

[0077] In certain embodiments, the disease is a metabolic disease.

[0078] In certain embodiments, the metabolic disease is selected from metabolic syndrome, diabetes, dyslipidemia, fatty liver disease, non-alcoholic steatohepatitis, obesity, and insulin resistance.

[0079] In certain embodiments, the diabetes is type II diabetes.

[0080] In certain embodiments, the dyslipidemia is hyperlipidemia.

[0081] Also provided is a method for achieving an effect in a patient, comprising administering to the patient a therapeutically effective amount of a compound disclosed above, wherein the effect is selected from the group consisting of reduced triglycerides, reduced cholesterol, and reduced hemoglobin A1c.

[0082] Further provided is the method disclosed above, wherein the cholesterol is selected from LDL cholesterol and VLDL cholesterol.

[0083] Further provided is the method disclosed above, wherein the triglyceride is selected from plasma triglycerides and liver triglycerides.

[0084] Also provided herein is a method for inhibiting MCT4, comprising contacting MCT4 with a compound disclosed herein.

[0085] Also provided herein are methods for achieving an effect in a patient, comprising administering to the patient a therapeutically effective amount of a compound disclosed herein, wherein the effect is selected from cognitive enhancement.

[0086] In certain embodiments, the MCT4-mediated disease is selected from proliferative inflammatory diseases.

[0087] Also provided is a method of modulating MCT4-mediated functions in a subject, comprising administering a therapeutically effective amount of a compound disclosed herein.

[0088] Additionally, pharmaceutical compositions comprising the compounds disclosed herein together with a pharmaceutically acceptable carrier are provided.

[0089] In certain embodiments, the pharmaceutical composition is formulated for oral administration.

[0090] In certain embodiments, the oral pharmaceutical composition is selected from a tablet and a capsule.

[0091] Abbreviations and Definitions As used herein, the following terms have the meanings indicated.

[0092] When used in a list of two or more items, the term "and / or" means that any one of the listed items may be used alone or in combination with any one or more of the listed items. For example, the phrase "A and / or B" is intended to mean either or both of A and B, i.e., A alone, B alone, or a combination of A and B. The phrase "A, B, and / or C" is intended to mean A alone, B alone, C alone, A and B in combination, A and C in combination, B and C in combination, or A, B, and C in combination.

[0093] When a range of values is disclosed and the notation "from n1... to n2" or "n1... to n2" is used, where n1 and n2 are numbers, unless otherwise specified, this notation is intended to include the numbers themselves and the range therebetween. The range may be an integer or continuous value therebetween, and includes the endpoints. For example, the range "2 to 6 carbon atoms" is intended to include carbon numbers 2, 3, 4, 5, and 6, since the number of carbon atoms is an integer unit. For comparison, "1 to 3 μM (micromolar)" is intended to include 1 μM, 3 μM, and all values between any number of significant digits (e.g., 1.255 μM, 2.1 μM, 2.9999 μM, etc.).

[0094] The term "about" as used herein is intended to modify the numerical value it modifies and indicates such value as being variable within a margin of error. When a specific margin of error is not specified, such as a standard deviation for an average value shown in a chart or table of data, the term "about" should be understood to mean a range that includes the specified value and a range that may be encompassed by rounding up or down to that numerical value, taking into account significant digits.

[0095] Asymmetric centers exist in the compounds disclosed herein. These centers are designated by the symbols "R" or "S," depending on the configuration of substituents around the asymmetric carbon atom. It should be understood that the present disclosure encompasses all stereochemical isomers, including diastereomeric, enantiomeric, and epimeric forms, as well as d- and 1-isomers, and mixtures thereof. Individual stereoisomers of the compounds can be prepared by synthesis from commercially available starting materials containing chiral centers, or by preparation and subsequent separation of a mixture of enantiomeric products, for example, by conversion to a mixture of diastereomers followed by separation or recrystallization, chromatographic techniques, direct separation of enantiomers on a chiral chromatographic column, or other suitable methods known in the art. Starting compounds of particular stereochemistry are commercially available or can be prepared and resolved by techniques known in the art. Additionally, the compounds disclosed herein may exist as geometric isomers. The present disclosure includes all cis, trans, syn, anti, entgegen (E), and zusammen (Z) isomers, as well as the appropriate mixtures thereof. Additionally, compounds may exist as tautomers; all tautomers are provided by the present disclosure. Furthermore, the compounds disclosed herein can exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents, such as water, ethanol, and the like. In general, solvated forms are considered equivalent to unsolvated forms.

[0096] As used herein, the term "disease" is generally intended to be synonymous with, and is used interchangeably with, the terms "disorder," "syndrome," and "condition" (in medical conditions), in that it refers to any abnormal condition of the human or animal body or one of its parts that impairs normal functioning and is typically manifested by characteristic signs and symptoms, and that results in a decrease in the lifespan or quality of life of the human or animal.

[0097] The term "combination therapy" refers to the administration of two or more therapeutic agents to treat a condition or disorder described in this disclosure. Such administration includes the co-administration of these therapeutic agents substantially simultaneously, such as in a single capsule having a fixed ratio of active ingredients or in multiple separate capsules for each active ingredient. In addition, such administration also includes the sequential use of each type of therapeutic agent. In either case, the treatment regimen will result in the beneficial effects of the drug combination in treating the condition or disorder described herein.

[0098] The term "MCT4 inhibitor" is used herein to mean a compound that exhibits an IC50 against MCT4 activity of about 100 μM or less, more typically about 50 μM or less, as measured in the MCT4 enzyme assay outlined below. IC50 is the concentration of an inhibitor that reduces the activity of an enzyme (e.g., MCT4) to half-maximal levels. Certain compounds disclosed herein have been found to exhibit inhibition against MCT4. In certain embodiments, the compound exhibits an IC50 against MCT4 of about 10 μM or less, as measured in the MCT4 binding assay described herein; in other embodiments, the compound exhibits an IC50 against MCT4 of about 5 μM or less; in yet other embodiments, the compound exhibits an IC50 against MCT4 of about 1 μM or less; in yet other embodiments, the compound will exhibit an IC50 against MCT4 of about 200 nM or less.

[0099] The phrase "therapeutically effective" is intended to qualify the amount of active ingredient used in treating a disease or disorder or achieving a clinical endpoint.

[0100] The term "therapeutically acceptable" refers to a compound (or salt, prodrug, tautomer, zwitterionic form, etc.) that is suitable for use in contact with the tissues of a patient without undue toxicity, irritation, or allergic response, is commensurate with a reasonable benefit / risk ratio, and is effective for its intended use.

[0101] As used herein, the terms "treat," "treating," or "treatment" refer to the administration of a therapeutic regimen to an individual who already exhibits at least one symptom of a disease or condition or who previously exhibited at least one symptom of a disease or condition. For example, "treating" can include alleviating, reducing, or ameliorating the symptoms of a disease or condition, preventing further symptoms, ameliorating the underlying metabolic cause of a symptom, inhibiting a disease or condition, e.g., preventing the onset of a disease or condition, relieving a disease or condition, causing regression of a disease or condition, alleviating the condition caused by a disease or condition, or halting the symptoms of a disease or condition. For example, the term "treating" with respect to a disorder refers to a reduction in the severity of one or more symptoms associated with that particular disorder. Thus, treating a disorder does not necessarily mean a reduction in the severity of all symptoms associated with the disorder, and does not necessarily mean a complete reduction in the severity of one or more symptoms associated with the disorder.

[0102] The term "patient" is generally synonymous with the term "subject" and includes all mammals, including humans. Examples of patients include humans, livestock such as cows, goats, sheep, pigs, and rabbits, and companion animals such as dogs, cats, rabbits, and horses. Preferably, the patient is a human.

[0103] While the compounds of the subject invention can be administered as the raw chemical, they can also be provided as pharmaceutical formulations. Thus, pharmaceutical formulations include one or more of the specific compounds disclosed herein, or one or more pharmaceutically acceptable salts, esters, prodrugs, amides, or solvates thereof, together with one or more pharmaceutically acceptable carriers thereof, and, optionally, one or more other therapeutic ingredients. The carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. Appropriate formulations will vary depending on the route of administration chosen. Any of the well-known techniques, carriers, and excipients may be used as suitable and as understood in the art. The pharmaceutical compositions disclosed herein can be manufactured by any method known in the art, for example, by conventional mixing, dissolving, granulating, dragee-making, preparing a homogenous mixture, emulsifying, encapsulating, entrapping, or compressing processes.

[0104] Formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous, intraarticular, and intramedullary), intraperitoneal, transmucosal, transdermal, rectal, and topical (including transdermal, buccal, sublingual, and ocular) administration, although the most suitable route may vary depending, for example, on the condition and disorder of the recipient. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Typically, these methods include the step of bringing into association a compound of the subject invention or a pharmaceutically acceptable salt, ester, amide, prodrug, or solvate thereof (the "active ingredient") with the carrier, which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing the active ingredients into association with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the mixture into the desired formulation.

[0105] Formulations of the compounds disclosed herein suitable for oral administration can be presented as discrete units such as capsules, cachets, or tablets, each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient can also be presented as a bolus, electuary, or paste.

[0106] Orally usable pharmaceutical formulations include tablets, push-fit capsules made of gelatin, and soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. Tablets can be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free-flowing form, such as a powder or granules, optionally mixed with a binder, inert diluent, or lubricant, surfactant, or dispersing agent. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. Tablets may optionally be coated or scored and may be formulated to provide slow or controlled release of the active ingredient therein. All formulations intended for oral administration should be in dosages suitable for such administration. Push-fit capsules can contain the active ingredient mixed with fillers such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate, and optionally, stabilizers. In soft capsules, the active compound can be dissolved or suspended in a suitable liquid, such as fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers may be added. Dragee cores are provided with a suitable coating. For this purpose, concentrated sugar solutions can be used, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to tablets or dragee coatings for identification or to characterize the dosage of various combinations of active compounds.

[0107] The compounds can also be formulated for parenteral administration by injection, e.g., bolus injection or continuous infusion. Injectable formulations can be provided in unit dosage form, e.g., in ampoules or multi-dose containers, with an added preservative. The compositions can take the form of suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and / or dispersing agents. The formulations can be provided in unit-dose or multi-dose containers, e.g., sealed ampoules and vials, and can be stored in powder form or lyophilized (lyophilized) condition, requiring only the addition of a sterile liquid carrier, e.g., saline or sterile pyrogen-free water, immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the type previously described.

[0108] Preparations for parenteral administration include aqueous and non-aqueous (oily) sterile injection solutions of the active compound (which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient); and aqueous and non-aqueous sterile suspensions that may contain suspending agents and thickening agents. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compound, allowing for the preparation of highly concentrated solutions.

[0109] In addition to the above-mentioned formulations, the compound can also be formulated as a depot preparation. Such long-acting preparations can be administered by implantation (for example, subcutaneously or intramuscularly) or intramuscular injection. Thus, for example, the compound can be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion exchange resins, or as a sparingly soluble derivative, for example, as a sparingly soluble salt.

[0110] For buccal or sublingual administration, the compositions may take the form of tablets, lozenges, pastilles, or gels formulated in a conventional manner. Such compositions may comprise the active ingredient in a flavored base such as sucrose and acacia or tragacanth.

[0111] The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, eg, containing conventional suppository bases such as cocoa butter, polyethylene glycol, or other glycerides.

[0112] Certain compounds disclosed herein can be administered topically, i.e., non-systemically. This includes topical application of the compounds disclosed herein to the epidermis or buccal cavity, as well as instillation of such compounds into the ear, eye, and nose, where the compounds do not significantly enter the bloodstream. In contrast, systemic administration refers to oral, intravenous, intraperitoneal, and intramuscular administration.

[0113] Suitable formulations for topical administration include liquid or semi-liquid formulations suitable for penetration through the skin to the site of inflammation, such as gels, liniments, lotions, creams, ointments, or pastes, as well as drops suitable for administration to the eye, ear, or nose. For topical administration, the active ingredient may comprise, for example, 0.001% to 10% w / w (weight percent) of the formulation. In certain embodiments, the active ingredient may comprise up to 10% w / w. In other embodiments, the active ingredient may comprise less than 5% w / w. In certain embodiments, the active ingredient may comprise 2% w / w to 5% w / w. In other embodiments, the active ingredient may comprise 0.1% to 1% w / w of the formulation.

[0114] For administration by inhalation, the compound can be conveniently delivered from an air inhaler, a nebulizer pressurized pack, or other suitable means for delivering an aerosol spray. The pressurized pack may contain a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Alternatively, for administration by inhalation or insufflation, the compound according to the present invention may be in the form of a dry powder composition, for example, a powder mix of the compound and a suitable powder base such as lactose or starch. The powder composition may be presented in unit dosage form, for example, in capsules, cartridges, gelatin, or blister packs, from which the powder can be administered by an inhaler or air inhaler.

[0115] Preferred unit dosage formulations are those containing an effective dose, as herein below recited, or an appropriate fraction thereof, of the active ingredient.

[0116] It should be understood that the foregoing formulations may include, in addition to the ingredients specifically named above, other agents conventional in the art having regard to the type of formulation in question; for example, those suitable for oral administration may contain flavoring agents.

[0117] The compounds can be administered orally or by injection at a dose of 0.1 to 500 mg / kg per day. The dose range for adults is generally 5 mg to 2 g per day. Tablets or other forms provided in individual units may conveniently contain an amount of one or more compounds effective in a unit containing such a dose or multiples thereof, e.g., 5 mg to 500 mg, usually about 10 mg to 200 mg.

[0118] The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending upon the host treated and the particular mode of administration.

[0119] The compound can be administered in various ways, for example, orally, topically, or by injection. The exact amount of compound administered to a patient is up to the attending physician. The specific dose level for a particular patient will depend on a variety of factors, including the activity of the particular compound used, age, body weight, general health, sex, diet, time of administration, route of administration, excretion rate, drug combination, the exact disorder being treated, and the severity of the indication or condition being treated. The route of administration may also vary depending on the condition and its severity.

[0120] In certain cases, it may be appropriate to administer at least one of the compounds described herein (or a pharmaceutically acceptable salt, ester, or prodrug thereof) in combination with another therapeutic agent. By way of example only, if hypertension is one of the side effects experienced by a patient upon receiving one of the compounds described herein, it may be appropriate to administer an antihypertensive agent in combination with the first therapeutic agent. Alternatively, by way of example only, the therapeutic effect of one of the compounds described herein may be enhanced by the administration of an adjuvant (i.e., an adjuvant may have minimal therapeutic benefit by itself, but when combined with another therapeutic agent, enhances the overall therapeutic benefit to the patient). Alternatively, by way of example only, the benefit experienced by a patient may be increased by administering one of the compounds described herein with another therapeutic agent (including a treatment regimen) that also has therapeutic benefit. By way of example only, in a treatment for diabetes that includes the administration of one of the compounds described herein, an enhanced therapeutic benefit may be achieved by also providing the patient with another diabetes therapeutic agent. In either case, regardless of the disease, disorder, or condition being treated, the overall benefit experienced by the patient may simply be the additive effect of the two therapeutic agents, or the patient may experience a synergistic benefit.

[0121] Accordingly, in another aspect, certain embodiments provide methods for treating an MCT4-mediated disorder in a human or animal subject in need of such treatment, comprising administering to the subject an amount of a compound disclosed herein in combination with at least one additional agent known in the art for treating the disorder effective to alleviate or prevent the disorder. In a related aspect, certain embodiments provide therapeutic compositions comprising at least one compound disclosed herein in combination with one or more additional agents for the treatment of an MCT4-mediated disorder.

[0122] Also provided herein are methods of treating a monocarboxylate transporter MCT4-mediated disorder in a subject in need thereof, comprising the sequential or co-administration of a compound disclosed herein with another therapeutic agent.

[0123] In certain embodiments, the therapeutic agent is a protein kinase inhibitor.

[0124] In certain embodiments, the protein kinase inhibitor is selected from Aurora B, EGFR, PLK-1, and CDK inhibitors.

[0125] In certain embodiments, the therapeutic agent is selected from antimetabolites, bcr-abl inhibitors, DNA damaging agents, EGFR inhibitors, microtubule stabilizing inhibitors, mitotic arrest inhibitors, S-phase inhibitors, and taxanes.

[0126] In certain embodiments, the therapeutic agent is a DNA damaging agent selected from alkylating agents, anthracyclines, antimetabolites, cross-linking agents, DNA replication inhibitors, intercalators, microtubule disrupting agents, PARP inhibitors, radiomimetic agents, radiosensitizers, strand breakers, and topoisomerase II inhibitors.

[0127] In certain embodiments, the therapeutic agent is one of the following: aminoglutethimide, amsacrine, anastrozole, asparaginase, valasertib, BCG, bicalutamide, bleomycin, buserelin, busulfan, campothecin, capecitabine, carboplatin, carmustine, chlorambucil, chloroquine, cisplatin, cladribine, clodronate, colchicine, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, doxorubicin, fluticasone, fluoxetine, fluoxetine-10 ... Norubicin, demethoxyviridine, dichloroacetate, ginestrol, diethylstilbestrol, docetaxel, doxorubicin, epirubicin, estradiol, estramustine, etoposide, everolimus, exemestane, filgrastim, fludarabine, fludrocortisone, fluorouracil, fluoxymesterone, flutamide, gemcitabine, genistein, goserelin, hydroxyurea, idarubicin, ifosfamide, imatinib Tinib, interferon, irinotecan, ironotecan, letrozole, leucovorin, leuprolide, levamisole, lomustine, lonidamine, mechlorethamine, medroxyprogesterone, megestrol, melphalan, mercaptopurine, mesna, metformin, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, nocodazole, olaparib, octreotide, oxaliplatin, paclitaxel, pamidronate The active ingredient is selected from the group consisting of acetaminophen, pentostatin, perifosine, plicamycin, porfimer, procarbazine, raltitrexed, rituximab, sorafenib, streptozocin, sunitinib, suramin, tamoxifen, temozolomide, temsirolimus, teniposide, testosterone, thioguanine, thiotepa, titanocene dichloride, topotecan, trastuzumab, tretinoin, vinblastine, vincristine, vindesine, and vinorelbine.

[0128] In certain embodiments, the therapeutic agent is selected from paracetamol, acetaminophen, pirfenidone, nintedanib, and non-hormonal contraceptives.

[0129] When used in cancer and neoplastic diseases, MCT4 inhibitors may be optimally used in conjunction with one or more of the following non-limiting examples of anti-cancer agents: (1) alkylating agents, including but not limited to cisplatin (PLATIN), carboplatin (PARAPLATIN), oxaliplatin (ELOXATIN), streptozocin (ZANOSAR), busulfan (MYLERAN), and cyclophosphamide (ENDOXAN); (2) antimetabolites, including but not limited to mercaptopurine (PURINETHOL), thioguanine, pentostatin (NIPENT), cytosine arabinoside (ARA-C), gemcitabine (GEMZAR), fluorouracil (CARAC), leucovorin (FUSILEV), and methotrexate (RHEUMATREX); (3) antimetabolites, including but not limited to vincristine (ONCO), (4) plant alkaloids and terpenoids, including but not limited to irinotecan (CAMPTOSAR), topotecan (HYCAMTIN), and etoposide (EPOSIN); (5) cytotoxic antibiotics, including but not limited to actinomycin D (COSMEGEN), doxorubicin (ADRIAMYCIN), bleomycin (BLENOXANE), and mitomycin (MITOSOL); (6) angiogenesis inhibitors, including but not limited to sunitinib (SUTENT) and bevacizumab (AVASTIN); and (7) tyrosine kinase inhibitors, including but not limited to imatinib (GLEEVEC), erlotinib (TARCEVA), lapatinib (TYKERB), and axitinib (INLYTA).

[0130] If a subject is suffering from or at risk of suffering from an inflammatory disease, the MCT4 inhibitor compounds described herein may optionally be used in any combination with one or more agents or methods for treating the inflammatory disease. Therapeutic agents / therapies for treating autoimmune diseases and / or inflammatory conditions include, but are not limited to, any of the following examples: (1) corticosteroids, including, but not limited to, cortisone, dexamethasone, and methylprednisolone; (2) ibuprofen, naproxen, acetaminophen, aspirin, fenoprofen (NALFON), flurbiprofen (ANSAID), ketoprofen, oxaprozin (DAYPRO), diazepam, benzodiazepine (ZO4), benzocaine (ZO6), benzodiazepine (ZO8), benzocaine (ZO9), benzodiazepine (ZO10), benzocaine (ZO11), benzocaine (ZO12), benzocaine (ZO13), benzocaine (ZO14), benzocaine (ZO15), benzocaine (ZO16), benzocaine (ZO17), benzocaine (ZO18), benzocaine (ZO19), benzocaine (ZO20), benzocaine (ZO21), benzocaine (ZO22), benzocaine (ZO23), benzocaine (ZO24), benzocaine (ZO25), benzocaine (ZO3), benzocaine (ZO34), benzocaine (ZO46), benzocaine (ZO47), benzocaine (ZO48), benzocaine (ZO49 ... Nonsteroidal anti-inflammatory drugs (NSAIDs), including lofenac sodium (VOLTAREN), diclofenac potassium (CATAFLAM), etodolac (LODINE), indomethacin (INDOCIN), ketorolac (TORADOL), sulindac (CLINORIL), tolmetin (TOLECTIN), meclofenamate (MECLOMEN), mefenamic acid (PONSTEL), nabumetone (RELAFEN), and piroxicam (FELDENE); (3 (3) immunosuppressants, including but not limited to methotrexate (RHEUMATREX), leflunomide (ARAVA), azathioprine (IMURAN), cyclosporine (NEORAL, SANDIMMUNE), tacrolimus, and cyclophosphamide (CYTOXAN); (4) CD20 blockers, including but not limited to rituximab (RITUXAN); (5) etanercept (ENBREL), infliximab (REMICADE), and adalimumab (6) interleukin-1 receptor antagonists, including but not limited to anakinra (KINERET); (7) interleukin-6 inhibitors, including but not limited to tocilizumab (ACTEMRA); (8) interleukin-17 inhibitors, including but not limited to AIN457; (9) Janus kinase inhibitors, including but not limited to tasocitinib; (10) SYK inhibitors, including but not limited to fostamatinib.

[0131] In certain embodiments, the method further comprises performing a non-chemical method of cancer treatment.

[0132] In certain embodiments, the method further comprises the administration of radiation therapy.

[0133] In certain embodiments, the method includes performing surgery, thermal ablation, focused ultrasound therapy, cryotherapy, or any combination thereof.

[0134] In either case, the multiple therapeutic agents (at least one of which is a compound disclosed herein) may be administered in any order, or simultaneously. If administered simultaneously, the multiple therapeutic agents may be provided in a single, unified form, or in multiple forms (by way of example only, as a single pill or as two separate pills). One of the therapeutic agents may be administered in multiple doses, or both may be administered in multiple doses. If not simultaneous, the timing between the multiple doses can be any period ranging from a few minutes to four weeks.

[0135] Also provided are methods for treating an MCT4-mediated disorder in a human or animal subject in need of such treatment, comprising administering to the subject an amount of a compound disclosed herein in combination with at least one additional agent known in the art for treating the disorder effective to alleviate or prevent the disorder. In a related aspect, certain embodiments provide therapeutic compositions comprising at least one compound disclosed herein in combination with one or more additional agents for treating an MCT4-mediated disorder.

[0136] Also provided are compounds and pharmaceutical compositions that inhibit glutaminase activity, particularly MCT4 activity, and are therefore useful for treating or preventing disorders associated with MCT4. The compounds and pharmaceutical compositions described herein selectively modulate MCT4 and are therefore useful for treating or preventing a variety of disorders associated with MCT4, including, but not limited to, proliferative and inflammatory diseases.

[0137] Accordingly, provided herein is a method for inhibiting the activity of the monocarboxylate transporter MCT4, or a variant thereof, in a biological sample, comprising contacting the biological sample with a compound disclosed herein.

[0138] Also provided herein is a method of inhibiting the activity of the monocarboxylate transporter MCT4, or a variant thereof, in a patient, comprising contacting the patient with a compound disclosed herein.

[0139] Also provided is a method of selectively inhibiting the activity of the monocarboxylate transporter MCT4, or a variant thereof, over the monocarboxylate transporter MCT1, or a variant thereof, in a patient, comprising administering to the patient a compound disclosed herein.

[0140] In certain embodiments, the inhibition is at least 100-fold selective for MCT4 over MCT1.

[0141] In certain embodiments, the compounds, salts, and pharmaceutical compositions described herein may be useful for treating or preventing cancer.

[0142] In certain embodiments, the compounds and salts described herein can be used to prevent or treat cancer, wherein the cancer is one of the following or variants thereof: acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), adrenocortical carcinoma, AIDS-related cancers (Kaposi's sarcoma and lymphoma), anal cancer, appendix cancer, atypical teratoid / rhabdoid tumor, basal cell carcinoma, cholangiocarcinoma (including extrahepatic cholangiocarcinoma), bladder cancer, bone cancer (including osteosarcoma and malignant fibrous histiocytoma), brain tumors (e.g., astrocytoma, brain and spinal cord tumors, brain stem glioma, central nervous system atypical teratoid / rhabdoid tumor). tumors, central nervous system embryonal tumors, craniopharyngioma, ependymomas, ependymomas, medulloblastomas, medulloepithelioma, intermediate pineal parenchymal tumors, supratentorial primitive neuroectodermal tumors, and pineoblastomas), breast cancer, bronchial tumors, Burkitt's lymphoma, carcinoid tumors, cancer of unknown primary origin, central nervous system (e.g., atypical teratoid / rhabdoid tumors, embryonal tumors, and lymphomas), cervical cancer, childhood cancer, chordoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma (mycosis fungoides and Sézary syndrome), cholangiocarcinoma, cholangiocarcinoma (extrahepatic) ), Ductal carcinoma in situ (DCIS), Embryonal tumors (central nervous system), Endometrial cancer, Ependymoblastoma, Ependymoma, Esophageal cancer, Nasal neuroblastoma, Ewing's sarcoma family tumors, Extracranial germ cell tumors, Extragonadal germ cell tumors, Extrahepatic bile duct cancer, Eye cancers (e.g., intraocular melanoma, retinoblastoma), Fibrous histiocytoma of bone (including malignant tumors and osteosarcomas), Gallbladder cancer, Gastric (stomach) cancer, Gastrointestinal carcinoid tumors, Gastrointestinal stromal tumors (GIST), Germ cell tumors (extracranial, extragonadal, ovarian), Gestational trophoblastic tumors, Glioma, Hairy cell leukemia, Head and neck cancer, Heart cancer, Hepatocellular (liver) carcinoma, Histiocytosis, Langerhans cell , Hodgkin's lymphoma, hypopharyngeal carcinoma, intraocular melanoma, islet cell tumors (endocrine, pancreatic), Kaposi's sarcoma, kidney (including renal cell), Langerhans cell histiocytosis, laryngeal cancer, leukemia (acute lymphoblastic (ALL), acute myeloid (AML), chronic lymphocytic (CLL), chronic myeloid (CML), hairy cell), lip and oral cancer, liver cancer (primary), lobular carcinoma in situ (LCIS), lung cancer (non-small cell and small cell), lymphoma (AIDS-related, Burkitt's, cutaneous T-cell (mycosis fungoides and Sézary syndrome), Hodgkin's, non-Hodgkin's, primary central nervous system (CNS), macroglobulinemia,Waldenstrom's, male breast cancer, malignant fibrous histiocytoma of bone and osteosarcoma, medulloblastoma, medulloepithelioma, melanoma (including intraocular (eye)), Merkel cell carcinoma, mesothelioma (malignant), metastatic squamous cell carcinoma of the neck with unknown primary, NU midline carcinoma, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, myeloid leukemia, chronic (CML), myeloid leukemia, acute (AML), myeloma and multiple myeloma, Myeloproliferative disorders (chronic), nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cavity cancer, oral cancer, lip, and oropharyngeal cancer, osteosarcoma and malignant fibrous histiocytoma of bone, ovarian cancer (e.g., epithelial, germ cell tumors, and low-grade malignant tumors), pancreatic cancer (including islet cell tumors), papillomatosis, paraganglioma, paranasal sinus and nasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, intermediate pineal parenchymal tumor, pineoblastoma, and supratentorial undifferentiated extraneuronal tumor Germinal tumors, pituitary tumors, plasma cell neoplasms / multiple myeloma, pleuropulmonary blastoma, gestational and breast cancer, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, renal cell (kidney) cancer, renal pelvis and ureter, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcomas (e.g., Ewing's sarcoma family of tumors, Kaposi's, soft tissue, uterine, etc.), Sezary syndrome, skin cancer (e.g., melanoma, Merkel cell carcinoma, non-melanoma), small cell lung cancer, small intestine cancer, soft tissue sarcoma, tonsillitis Squamous cell carcinoma, squamous cell neck cancer of unknown primary, metastatic, gastric (stomach) cancer, supratentorial primitive neuroectodermal tumor, T-cell lymphoma (skin, mycosis fungoides and Sezary syndrome), testicular cancer, pharyngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor (gestational), unusual childhood cancer of unknown primary, ureter and renal pelvis, transitional cell carcinoma, urethral cancer, uterine cancer, endometrium, uterine sarcoma, Waldenstrom's macroglobulinemia or Wilms' tumor.

[0143] In certain embodiments, the cancer to be treated is T-cell specific, such as T-cell lymphoma and lymphoblastic T-cell leukemia.

[0144] In certain embodiments, the methods described herein are used to treat a disease state, comprising administering a therapeutically effective amount of a compound as described herein to a subject in need thereof, wherein the condition is cancer that has developed resistance to chemotherapeutic agents and / or ionizing radiation.

[0145] In certain embodiments, the compounds, salts, and pharmaceutical compositions described herein are useful for treating or preventing inflammatory diseases.

[0146] In certain embodiments, the compounds and salts described herein can be used to prevent or treat an inflammatory disease, wherein the inflammatory disease is one of the following or variants thereof: acid-induced lung injury, acne (PAPA), acute respiratory distress syndrome, Addison's disease, adrenal hyperplasia, adrenal insufficiency, aging, AIDS, alcoholic hepatitis, alcoholic liver disease, allergen-induced asthma, allergic bronchopulmonary aspergillosis, allergic conjunctivitis, alopecia, Alzheimer's disease, amyloidosis, amyotrophic lateral sclerosis, angina pectoris, angioedema, anhidrosis. Ectodermal hypoplasia (e.g., with immunodeficiency), ankylosing spondylitis, anterior segment inflammation, antiphospholipid syndrome, aphthous stomatitis, appendicitis, asthma, atherosclerosis, atopic dermatitis, autoimmune diseases, autoimmune hepatitis, bee sting-induced inflammation, Behçet's disease, Bell's palsy, Berylliosis, Blau syndrome, bone pain, bronchiolitis, burns, bursitis, cardiomegaly, carpal tunnel syndrome, catabolic disorders, cataracts, cerebral aneurysms, chemical irritant-induced inflammation, chorioretinitis, chronic heart failure, chronic lung disease of prematurity, chronic obstructive pulmonary disease, colitis, complex regional pain syndrome, connective tissue diseases, corneal ulcers, Crohn's disease, cryopyrin-associated periodic syndrome, cryptococcosis, cystic fibrosis, interleukin-1 receptor agonist deficiency, dermatitis, dermatitis endotoxemia, dermatomyositis, endometriosis, endotoxemia, epicondylitis, erythroblastopenia, familial amyloidotic polyneuropathy, familial cold urticaria, familial Mediterranean fever, fetal growth retardation, glaucoma, glomerular disease, glomerulonephritis, gout, gouty arthritis, graft-versus-host disease, intestinal disease, head trauma, headache, hearing loss, heart disease, hemolytic anemia, Henoch-Scholein purpura, hepatitis, hereditary periodic fever syndromes, shingles, and herpes simplex HIV-1, Huntington's disease, hyaline membrane disease, hyperammonemia, hypercalcemia, hypercholesterolemia, recurrent fever with hyperimmunoglobulinemia D, aplastic anemia and other anemias, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura, incontinentia pigmenti, infectious mononucleosis, inflammatory bowel disease, inflammatory lung disease, inflammatory neuropathy, inflammatory pain, insect sting-induced inflammation, iritis, ischemia / reperfusion, juvenile rheumatoid arthritis, keratitis, kidney disease, kidney damage due to parasitic infection, kidney transplant rejection prevention, leptospirosis, Löffler's syndrome, lung damage, lupus, lupus nephritis, meningitis, mesothelioma,Mixed connective tissue disease, Muckle-Wells syndrome (amyloidosis with urticaria and hearing loss), multiple sclerosis, muscle wasting, muscular dystrophy, myasthenia gravis, myocarditis, mycosis fungoides, myelodysplastic syndrome, myositis, sinusitis, necrotizing enterocolitis, neonatal-onset multisystem inflammatory disease (NOMID), nephrotic syndrome, neuritis, neurological disorders, non-allergenic asthma, obesity, ocular allergy, optic neuritis, organ transplant, osteoarthritis, otitis media, Paget's disease, pain, pancreatitis, Parkinson's disease, pemphigus, pericarditis, periodic fever, periodontitis, whooping cough, perineal or peritoneal endometriosis, pharyngitis and adenitis (PFAPA syndrome), plant irritant-induced inflammation, Pneumocystis infection, pneumonia, pneumonitis, poison ivy ivy) / urushiol oil-induced inflammation, polyarteritis nodosa, polycystic kidney disease, polychondritis, polycystic kidney disease, polymyositis, psoriasis, psychosocial stress disorders, lung disease, pulmonary fibrosis, pulmonary hypertension, pyoderma gangrenosum, suppurative aseptic arthritis, kidney disease, retinal disease, rheumatic diseases, rheumatoid arthritis, rheumatic carditis, sarcoidosis, seborrhea, sepsis, severe pain, sickle cell anemia, sickle cell disease, silica-induced disease, Sjogren's disease syndrome, skin diseases, sleep apnea, spinal cord injury, Stevens-Johnson syndrome, stroke, subarachnoid hemorrhage, sunburn, systemic sclerosis (scleroderma), temporal arteritis, tenosynovitis, thrombocytopenia, thyroiditis, tissue transplant, TNF receptor-associated periodic syndrome (TRAPS), toxoplasmosis, transplant, traumatic brain injury, tuberculosis, type 1 diabetes, type 2 diabetes, ulcerative colitis, urticaria, uveitis, Wegener's granulomatosis, and weight loss.

[0147] Thus, in another aspect, certain embodiments provide a method of treating a monocarboxylic acid transporter MCT4-mediated disorder in a subject in need thereof, comprising administering to the patient a therapeutically effective amount of a compound as disclosed herein.

[0148] In certain embodiments, the subject is a human.

[0149] In certain embodiments, the subject is in a fed state.

[0150] In certain embodiments, the subject is in a fasted state.

[0151] In certain embodiments, the monocarboxylate transporter MCT4-mediated disorder is selected from an inflammatory disorder and a proliferative disorder.

[0152] In certain embodiments, the monocarboxylate transporter MCT4-mediated disorder is a proliferative disorder.

[0153] In certain embodiments, the proliferative disorder is cancer.

[0154] In certain embodiments, the cancer is selected from the group consisting of: adenocarcinoma, adult T-cell leukemia / lymphoma, bladder cancer, blastoma, bone cancer, breast cancer, brain tumor, carcinoma, myeloid sarcoma, cervical cancer, colon cancer, esophageal cancer, gastrointestinal cancer, glioblastoma multiforme, glioma, gallbladder cancer, gastric cancer, head and neck cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma, intestinal cancer, kidney cancer, laryngeal cancer, leukemia, lung cancer, lymphoma, liver cancer, small cell lung cancer. , non-small cell lung cancer, mesothelioma, multiple myeloma, eye cancer, optic nerve tumor, oral cancer, ovarian cancer, pituitary tumor, primary central nervous system lymphoma, prostate cancer, pancreatic cancer, pharyngeal cancer, renal cell carcinoma, rectal cancer, sarcoma, skin cancer, spinal cord tumor, small intestine cancer, gastric cancer, T-cell lymphoma, testicular cancer, thyroid cancer, pharyngeal cancer, genitourinary cancer, urothelial carcinoma, uterine cancer, vaginal cancer, and Wilms' tumor.

[0155] In certain embodiments, the monocarboxylate transporter MCT4-mediated disorder is an inflammatory disorder.

[0156] In certain embodiments, the inflammatory disorder is selected from Crohn's disease, ulcerative colitis, idiopathic pulmonary fibrosis, muscular dystrophy, rheumatoid arthritis, and systemic sclerosis (scleroderma). In certain embodiments, the inflammatory disease is idiopathic pulmonary fibrosis.

[0157] In certain embodiments, the therapeutically effective amount is from about 30 mg to about 200 mg. In certain embodiments, the therapeutically effective amount is from about 30 mg to about 80 mg. In certain embodiments, the therapeutically effective amount is selected from 50 mg, 75 mg, 100 mg, 150 mg, and 200 mg.

[0158] Also provided herein are compounds as disclosed herein for use in human therapy.

[0159] Further provided herein is a compound as disclosed herein, e.g., as disclosed in any of the above embodiments and paragraphs relating to methods of treatment, for use in treating a monocarboxylate transporter MCT4-mediated disorder.

[0160] Also provided herein is the use of a compound as disclosed herein, e.g., as disclosed in any of the above embodiments and paragraphs relating to methods of treatment, for the manufacture of a medicament for the treatment of a monocarboxylic acid transporter MCT4-mediated disorder.

[0161] Metabolic syndrome (also known as metabolic syndrome X) is characterized by the presence of at least three of the following symptoms: insulin resistance; abdominal fat—defined as a waist of 40 inches or more in men and 35 inches or more in women; high blood glucose levels—at least 110 milligrams per deciliter (mg / dL) after fasting; high triglycerides—at least 150 mg / dL in the bloodstream; low HDL levels—less than 40 mg / dL; a thrombophilic state (e.g., high blood levels of fibrinogen or plasminogen activator inhibitor); or blood pressure of 130 / 85 mmHg or higher. Metabolic syndrome has been associated with other conditions, such as obesity, high blood pressure, and high LDL cholesterol levels, all of which are risk factors for cardiovascular disease. For example, metabolic syndrome has been strongly associated with atherosclerosis. Individuals with metabolic syndrome are also more likely to develop type 2 diabetes, polycystic ovary syndrome (PCOS) in women, and prostate cancer in men.

[0162] As mentioned above, insulin resistance manifests in several ways, including type 2 diabetes, the condition most clearly associated with insulin resistance. Compensatory hyperinsulinemia helps maintain normal glucose levels for decades before overt diabetes develops. Eventually, pancreatic beta cells are unable to overcome insulin resistance through hypersecretion. When glucose levels rise, diabetes can be diagnosed. Patients with type 2 diabetes remain hyperinsulinemic until the disease progresses. As mentioned above, insulin resistance can also be correlated with hypertension. Half of patients with essential hypertension are insulin resistant and hyperinsulinemic, and there is evidence that blood pressure is related to the degree of insulin resistance. Hyperlipidemia is also associated with insulin resistance. The lipid profile of patients with type 2 diabetes includes elevated serum very-low-density lipoprotein cholesterol and triglyceride levels and, occasionally, decreased low-density lipoprotein cholesterol levels. Insulin resistance has been observed in individuals with low high-density lipoprotein levels. Insulin levels are also related to very low density lipoprotein synthesis and plasma triglyceride levels.

[0163] Accordingly, a method for treating insulin resistance in a subject is also disclosed, comprising the steps of selecting a subject in need of treatment for insulin resistance; and administering to the subject an effective amount of a compound that inhibits MCT4.

[0164] Specific diseases treated by the compounds, compositions, and methods disclosed herein are diseases mediated at least in part by MCT4. Accordingly, the following are disclosed herein: a method for reducing glycogen accumulation in a subject; a method for increasing HDL or HDLc, reducing LDL or LDLc, shifting LDL particle size from small dense to normal LDL, lowering VLDL, lowering triglycerides, or inhibiting cholesterol absorption in a subject; a method for reducing insulin resistance, enhancing glucose utilization, or lowering blood pressure in a subject; a method for reducing visceral fat in a subject; a method for reducing serum transaminases in a subject; or a method for treating a disease; all of which comprise administering a therapeutic amount of a compound described herein to a patient in need thereof. In another embodiment, the disease to be treated may be a metabolic disease. In yet another embodiment, the metabolic disease may be selected from the group consisting of obesity, diabetes, particularly type 2 diabetes, hyperinsulinemia, impaired glucose tolerance, metabolic syndrome X, dyslipidemia, hypertriglyceridemia, hypercholesterolemia, and fatty liver. In other embodiments, the disease to be treated may be selected from the group consisting of cardiovascular diseases, such as vascular disease, atherosclerosis, coronary heart disease, cerebrovascular disease, heart failure, and peripheral vascular disease. In a preferred embodiment, the method does not induce or maintain a hypoglycemic state.

[0165] In addition to being useful for human treatment, certain compounds and formulations disclosed herein may also be useful for the veterinary treatment of companion animals, exotic animals, and farm animals, including mammals, rodents, etc. More preferred animals include horses, dogs, and cats. [Example]

[0166] The present invention is further illustrated by the following examples.

[0167] Compound synthesis The compounds and salts can be prepared using methods exemplified in the general synthetic schemes and experimental procedures detailed below. The general synthetic schemes and experimental procedures are offered for illustrative purposes and are not intended to be limiting. The starting materials used to prepare the compounds and salts described herein are either commercially available or can be prepared using routine methods known in the art.

[0168] Example 1: 2-([1-[2-(azetidin-1-yl)phenyl]-5-(3-cyclobutoxyphenyl)-1H-pyrazol-3-yl]methoxy)-2-methylpropanoic acid [ka] The compound of structural formula I (ie, compound 1) was synthesized according to the steps in Scheme 1. Scheme 1 [ka]

[0169] Step 1: [ka] A mixture of 1-(3-hydroxyphenyl)ethan-1-one (20 kg), bromocyclobutane (1.3 eq.), and CsCO (1.5 eq.) in DMF (5 vol.) was stirred at 80±5°C for 16 h. The reaction vessel was charged with water (15 vol.) and methyl tert-butyl ether (MTBE, 15 vol.). The organic layer was separated and washed twice with 20% brine (5 vol.), then concentrated to approximately 5 vol. and solvent-exchanged three times with methanol to approximately 5 vol. The crude methanol solution containing compound 7 was used in the next step without further purification.

[0170] Step 2: [ka] To the crude methanol solution (5 volumes) from Step 1, sodium methoxide (2.0 eq.) and dimethyl oxalate (1.5 eq.) were added. The reaction vessel was stirred at 30±5°C for 16 hours. Upon completion, the reaction mixture was cooled to a temperature of 0-10°C. The pH was adjusted to 2-3 with 4.0 M HCl in methanol, and the crude solution containing compound 2 was used in the next step without further purification.

[0171] Step 3: [ka] To the reaction vessel containing the crude methanol solution from Step 2 was added 2-bromophenylhydrazine hydrochloride (1.0 eq.). The reaction mixture was then stirred at 60±5°C for 10 hours and then cooled to 5-15°C. The resulting solid was filtered, washed with methanol (1 vol), and then slurried in water (20 vol). The solid was filtered again and washed with water (2 vol). The product was dried in an oven at 50°C to give 54.9 kg of compound 3 (88% yield for Steps 1-3).

[0172] Step 4: [ka] Compound 3 (4.0 kg; 1.0 eq.) was added to THF (4 vol.), followed by methanol (0.4 vol.) in a reaction vessel that was cooled to 10-25°C. Sodium borohydride (1.2 eq.) was added, and the reaction was stirred at 10-25°C for 16 h. Upon completion, the reaction was cooled to 0-10°C, and the pH was adjusted to 3-5 with 0.5 M HCl, at which point the product precipitated from solution. The mixture was stirred at 5-15°C for an additional 1-2 h. The product was collected by filtration, washed with water, and dried in an oven at 50°C to give 3.5 kg of compound 4 (85% yield).

[0173] Step 4 was repeated on a 5x scale as follows: Compound 3 (20.0 kg; 1.0 eq.) was added to THF (6.0 vol.), followed by additional methanol (0.4 vol.) in the reaction vessel, which was cooled to 25±5°C. Sodium borohydride (1.2 eq.) was added, and the reaction was stirred at 25±5°C for 16 h. Upon completion, the reaction was cooled to 0-10°C and the pH was adjusted to 3-5 with 0.5 M HCl, at which point the product precipitated from solution. The mixture was stirred at 5-15°C for an additional 1-2 h. The product was collected by filtration, washed with water, and dried in an oven at 50°C to give 18.2 kg of compound 4 in 99.4% purity and 97% isolated yield.

[0174] Step 5: [ka] Compound 4 (2.97 kg; 1.0 eq.) was added to THF (10 vol.) under a nitrogen atmosphere with stirring. Xantphos (0.11 eq.), Pd(OAc)2 (0.11 eq.), and t-BuONa (2.0 eq.) were added to the reaction vessel. Azetidine (2.5 eq.) was added, and the resulting mixture was stirred at 25-35°C for 20 h. The vessel was charged with MTBE (20 vol.) and water (20 vol.). The organic layer was separated, washed with 5% aqueous NH4Cl and 5% aqueous NaCl, and filtered through Celite®. The resulting organic layer was concentrated and solvent-exchanged with ethyl acetate (10 vol.). The resulting solution was stirred at 70-80°C for 30 min, then cooled to 15-20°C and stirred for an additional 1-2 h. The solid was collected by filtration, washed with ethyl acetate (0.5 vol), and dried in an oven at 50° C. to give 1.7 kg of compound 5 in 60% yield. Reproduction with 18.0 kg of compound 4 gave 10.8 kg of compound 5 with a purity of 98.8% and an isolated yield of 63.8%.

[0175] Step 6 [ka] To a solution of compound 5 (700.0 g, 1 eq.) in DMF (8 vol.) was added isopropyl 2-bromo-2-methylpropanoate (8.0 eq.). The reaction mixture was cooled to -60°C to -50°C using liquid nitrogen. A solution of potassium bis(trimethylsilyl)amide (KHMDS, 5.0 eq.) in THF was added dropwise over 1.5 h; the reaction was then stirred for 30–50 min. Water (10 vol.) was added to the reaction vessel. The organic phase was collected, washed twice with NaCl (15%) (5 vol.), and concentrated. The crude compound 6 (70% yield) was used directly in the next step without further purification.

[0176] Step 7 [ka] Crude compound 6 (1950.0 g, 1 eq.) from Step 6 was added to a reaction vessel containing 30% (aq.) KOH (10 vol., 20 eq.) and methanol (5 vol.). The reaction was stirred at 50±5°C for 16 h and then cooled to 15–30°C. The solution was washed twice with MTBE (10 vol.), after which the aqueous layer was separated and filtered through Celite®. The pH of the resulting aqueous phase was adjusted to 3–5 with 2.0 M HCl. The solution was cooled to 5–10°C, stirred for 2–3 h, and then filtered. The filter cake was dissolved in THF (10 vol.), and mercaptosilica gel (300 g, 15% w / w) was added to the solution. The resulting mixture was heated to 50–60°C, stirred for 3 h, and then filtered. The solution was concentrated, and the crude product was dissolved in ethyl acetate (10% v / v). The resulting solution was heated to 70–80° C. and stirred for 3–4 h, then cooled to 0–10° C. The solid was collected by filtration, washed with ethyl acetate (0.5 vol), and then dried in an oven at 50° C. to give compound 1 (1.4 kg, 58% yield) as an off-white solid. 1H NMR(C2D6OS)δ 12.66(s,1H),7.20(dd,2H),6.98(t,2H),6.73(m,3H),6.60(d,1H),6.53(d,1H),4.45(s,2H),4.32(t,1H), 3.48(dd,4H),3.32(s,3H),2.27(d,2H),2.05(m,2H),1.93(m,2H),1.73(m,1H),1.56(m,1H),1.415(s,6H).

[0177] Example 2: Large-scale synthesis of 2-([1-[2-(azetidin-1-yl)phenyl]-5-(3-cyclobutoxyphenyl)-1H-pyrazol-3-yl]methoxy)-2-methylpropanoic acid and tris salt 1. Synthesis of Compound 1 Compound 1 was synthesized according to the steps in Scheme 1. Scheme 1 [ka]

[0178] Step 1: [ka] A mixture of 1-(3-hydroxyphenyl)ethan-1-one (20 kg), bromocyclobutane (1.3 eq.), and CsCO (1.5 eq.) in DMF (5 vol.) was stirred at 80±5°C for 16 h. The reaction vessel was charged with water (15 vol.) and methyl tert-butyl ether (MTBE, 15 vol.). The organic layer was separated and washed twice with 20% brine (5 vol.), then concentrated to approximately 5 vol. and solvent exchanged three times with methanol to approximately 5 vol. The crude methanol solution containing compound 7 was used in the next step without further purification.

[0179] Step 2: [ka] To the crude methanol solution from Step 1 (5 volumes) was added sodium methoxide (2.0 eq.) and dimethyl oxalate (1.5 eq.). The reaction vessel was stirred at 30±5°C for 16 hours. Upon completion, the reaction mixture was cooled to a temperature between 0 and 10°C. The pH was adjusted to 2-3 with 4.0 M HCl in methanol, and the crude solution containing compound 2 was used in the next step without further purification.

[0180] Step 3: [ka] To the reaction vessel containing the crude methanol solution from Step 2 was added 2-bromophenylhydrazine hydrochloride (1.0 eq.). The reaction mixture was then stirred at 60±5°C for 10 hours and then cooled to 5-15°C. The resulting solid was filtered, washed with methanol (1 vol), and then slurried in water (20 vol). The solid was filtered again and washed with water (2 vol). The product was dried in an oven at 50°C to give 54.9 kg of compound 3 (88% yield for Steps 1-3).

[0181] Step 4: [ka] Compound 3 (4.0 kg; 1.0 eq.) was added to THF (4 vol.), followed by methanol (0.4 vol.) in a reaction vessel that was cooled to 10-25°C. Sodium borohydride (1.2 eq.) was added, and the reaction was stirred at 10-25°C for 16 h. Upon completion, the reaction was cooled to 0-10°C, and the pH was adjusted to 3-5 with 0.5 M HCl, at which point the product precipitated from solution. The mixture was stirred at 5-15°C for an additional 1-2 h. The product was collected by filtration, washed with water, and dried in an oven at 50°C to give 3.5 kg of compound 4 (85% yield).

[0182] Step 4 was repeated on a 5x scale as follows: Compound 3 (20.0 kg; 1.0 eq.) was added to THF (6.0 vol.), followed by additional methanol (0.4 vol.) in the reaction vessel, which was cooled to 25±5°C. Sodium borohydride (1.2 eq.) was added, and the reaction was stirred at 25±5°C for 16 h. Upon completion, the reaction was cooled to 0-10°C and the pH was adjusted to 3-5 with 0.5 M HCl, at which point the product precipitated from solution. The mixture was stirred at 5-15°C for an additional 1-2 h. The product was collected by filtration, washed with water, and dried in an oven at 50°C to give 18.2 kg of compound 4 in 99.4% purity and 97% isolated yield.

[0183] Step 5: [ka] Compound 4 (2.97 kg; 1.0 eq.) was added to THF (10 vol.) under a nitrogen atmosphere with stirring. Xantphos (0.11 eq.), Pd(OAc)2 (0.11 eq.), and t-BuONa (2.0 eq.) were added to the reaction vessel. Azetidine (2.5 eq.) was added, and the resulting mixture was stirred at 25-35°C for 20 h. The vessel was charged with MTBE (20 vol.) and water (20 vol.). The organic layer was separated, washed with 5% aqueous NH4Cl and 5% aqueous NaCl, and then filtered through Celite®. The resulting organic layer was concentrated and solvent exchanged with ethyl acetate (10 vol.). The resulting solution was stirred at 70-80°C for 303 min, then cooled to 15-20°C and stirred for an additional 1-2 h. The solid was collected by filtration, washed with ethyl acetate (0.5 vol), and dried in an oven at 50° C. to give 1.7 kg of compound 5 in 60% yield. Reproduction with 18.0 kg of compound 4 gave 10.8 kg of compound 5 with a purity of 98.8% and an isolated yield of 63.8%.

[0184] Step 6 [ka] To a solution of compound 5 (700.0 g, 1 eq.) in DMF (8 vol.) was added isopropyl 2-bromo-2-methylpropanoate (8.0 eq.). The reaction mixture was cooled to -60°C to -50°C using liquid nitrogen. A solution of potassium bis(trimethylsilyl)amide (KHMDS, 5.0 eq.) in THF was added dropwise over 1.5 h; the reaction was then stirred for 30–50 min. Water (10 vol.) was added to the reaction vessel. The organic phase was collected, washed twice with NaCl (15%) (5 vol.), and concentrated. The crude compound 6 (70% yield) was used directly in the next step without further purification.

[0185] Step 7 [ka] Crude compound 6 (1950.0 g, 1 eq.) from Step 6 was added to a reaction vessel containing 30% (aq.) KOH (10 vol., 20 eq.) and methanol (5 vol.). The reaction was stirred at 50±5°C for 16 h and then cooled to 15–30°C. The solution was washed twice with MTBE (10 vol.), after which the aqueous layer was separated and filtered through Celite®. The pH of the resulting aqueous phase was adjusted to 3–5 with 2.0 M HCl. The solution was cooled to 5–10°C, stirred for 2–3 h, and then filtered. The filter cake was dissolved in THF (10 vol.), and mercaptosilica gel (300 g, 15% w / w) was added to the solution. The resulting mixture was heated to 50–60°C, stirred for 3 h, and then filtered. The solution was concentrated, and the crude product was dissolved in ethyl acetate (10% v / v). The resulting solution was heated to 70–80° C. and stirred for 3–4 h, then cooled to 0–10° C. The solid was collected by filtration, washed with ethyl acetate (0.5 vol), and then dried in an oven at 50° C. to give compound 1 (1.4 kg, 58% yield) as an off-white solid.

[0186] This reaction was repeated on a larger scale as follows: Crude compound 6 (8.4 kg, 1 eq.) from step 6 was added to a reaction vessel containing 30% (aq.) KOH (10 vol., 20 eq.) and methanol (5 vol.). After the same procedure as above, 8.76 kg of compound 1 was obtained with a purity of 97.7% and a 67% yield over two steps.

[0187] 2. Use of a different propanoate ester in Step 6 The propanoic acid ester in step 6 was varied and tested. The results are shown in Table 1. [ka]

[0188] [Table 1]

[0189] [Table 2]

[0190] As shown in Table 1, starting materials using the isopropyl ester performed better than t-butyl or methyl esters. In addition, the use of the isopropyl ester limits the potential for the formation of dimer (or bis-adduct) by-products detected when the methyl ester was used in Run 13. The use of the isopropyl ester also allows for the use of solvents other than DMF / NaH. The reaction also proceeded much faster than when using DMF / NaH.

[0191] 3. Formation of the Tris Salt of Compound 1 [ka] Compound 1 (8.4 kg, 1 eq.) was added to a reaction vessel containing THF (42 L, 5.0 vol.), charged with mercaptosilica gel (420 g, 5 wt.%), and then stirred and heated at 50–60°C for 3 hours. The reaction mixture was sampled, concentrated, and subjected to ICP analysis to determine the residual Pd concentration (approximately 15 ppm). The reaction mixture was filtered, and acetone (168 L, 20.0 vol.) and aqueous tris(hydroxymethyl)aminomethane ("Tris", 1.01 eq.) solution (4.2 L, 0.5 vol.) were added. The reaction mixture was stirred at 20–30°C for 5–10 minutes to dissolve most of the solids, and stirring was continued at 20–30°C for approximately 20 hours to precipitate the Tris salt of compound 1. The precipitate was filtered, washed with acetone (4.2 L, 0.5 vol), and dried under vacuum at 60° C. to give 8.4 kg of Compound 1 Tris in 99.5% purity and 79.2% yield, which had 1690 ppm residual acetone and 962 ppm residual THF.

[0192] Example 3: Salt Formulations method Polarized light microscopy (PLM) was performed using an Olympus BX60 polarized light microscope equipped with an Olympus DP70 camera.

[0193] Powder X-ray diffraction ("XRPD") was performed using a PANalytical X'Pert Pro diffractometer with Ni-filtered Cu Ka (45 kV / 40 mA) radiation and a step size of 0.02° 2θ and an X'celerator™ RTMS (Real Time Multi-Strip) detector. Incident beam configuration: fixed divergence slit (0.25°), 0.04 rad Soller slit, anti-scatter slit (0.25°), and 10 mm beam mask. Diffracted beam configuration: fixed divergence slit (0.25°) and 0.04 rad Soller slit. Samples were mounted flat on zero-background Si wafers.

[0194] Differential scanning calorimetry ("DSC") was performed using a TA Instruments Q100 differential scanning calorimeter equipped with an autosampler and a refrigerated cooling system under a 40 mL / min N purge. DSC thermograms were acquired in crimped Al pans at 15°C / min.

[0195] Thermogravimetric analysis ("TGA") was performed using a TA Instruments Q500 thermogravimetric analyzer in Pt or Al pans at 15°C / min under a 40 mL / min N2 purge.

[0196] Thermogravimetric analysis with IR exhaust gas detection (TGA-IR) was performed using a TA Instruments Q5000 thermogravimetric analyzer interfaced with a Nicolet 6700 FT-IR spectrometer (Thermo Electron) equipped with an external TGA-IR module equipped with a gas flow cell and a DTGS detector. TGA was performed in Pt or Al pans with a N2 flow of 60 mL / min and a heating rate of 15 °C / min. IR spectra were taken at 4 cm time points. -1 A resolution of 100 s and 32 scans were collected.

[0197] 1 H NMR spectra were collected on an Agilent DD2 500MH spectrometer referenced to TMS. Samples were dissolved in DMSO-d6.

[0198] Ion chromatography (IC) was performed on a Dionex ICS-5000. Column: Dionex IonPac CS12 4 x 250 mm; Detection: Suppressed conductivity, CERS 500 with a suppressor current of 59 mA; Eluent (20 mM methanesulfonic acid) 1.0 mL / min.

[0199] Preliminary salt formation experiment using the compound of Example 1 The salt experiments of Example 1 were performed using the following general procedure. Approximately 20 mg of the compound of Example 1 was weighed into individual vials. 200 μL or 1000 μL of solvent was added to each vial along with a stoichiometric amount of counterion. The resulting solution / suspension / gums were stirred while cycling between 40°C and 5°C for two days (TC1). After TC1, the solvent in the gum / solution was evaporated under reduced pressure, and 200 μL of solvent was re-aliquoted into the vials. The resulting solution / suspension / gums were stirred again while cycling between 40°C and 5°C for two days (TC2). The solutions / gums / gels were rapidly cooled to 4°C and held at 4°C for two days (RC). The solvent was then allowed to evaporate at ambient temperature for seven days (SEV). Samples were examined for birefringence at each step by PLM, and if present, were isolated, analyzed, and grouped by FT-Raman and / or XRPD. Representative samples from each group were also characterized by DSC, and samples with promising DSC results were further characterized by TGA-IR, XRPD, HNMR, and / or IC.

[0200] [Table 3]

[0201] Compound 1 Tris salt Form A (4088.62 mg) was transferred to a 125 mL Erlenmeyer flask, and 70 mL of solvent (95:5 acetone:water v / v) was added. The mixture was stirred magnetically. After 5 minutes of stirring, Compound 1 Tris salt Form B seed (53.19 mg), generated from the primary polymorph screen described above, was added and stirring continued. After 15 minutes of stirring, the sample solid became immobile and the stir bar stopped working. The flask was shaken by hand to break up clumps. A small aliquot was taken and analyzed by PXRD, which revealed the solid to be Form B. The remaining solid was isolated using a Büchner funnel with Whatman #1 filter paper. The solid was washed with 20 mL of acetone chilled to -20 °C. The funnel containing the isolated solid was covered with Kimwipes and allowed to dry on the filter under vacuum overnight. NMR analysis indicated a 1:1 molar ratio of Compound 1 to Tris salt. The chromatographic purity was 99.8% (254 nm).

[0202] After drying for approximately 14 hours, a solid was isolated (3804.50 mg, 93% yield). PXRD confirmed the sample to be Form B Tris salt with no detectable Form A Tris salt and no detectable parent compound 1. The sample showed negligible weight loss prior to thermal decomposition by TGA.

[0203] Characterization of Form A Tris Salt During salt testing, one crystalline Tris salt was isolated from six runs. This hit was designated Form A Tris salt. The remaining runs yielded either the parent Form A or an amorphous / gum. Thermal analysis indicated that Form A Tris was an unsolvated form.

[0204] DSC data showed a melting endotherm with an onset at 147.4 °C. TGA-IR analysis showed negligible water weight loss (0.3%) between 25 and 150 °C, indicating that the salt was unsolvated. 1 H-NMR showed the stoichiometry to be mono-salt (API / CI ratio 1:1).

[0205] Characterization of Form B Tris Salt Form B Tris salt is an off-white crystalline powder consisting of small particles. PXRD showed relatively sharp diffraction peaks between 2 and 40°2θ, consistent with a crystalline material. DSC analysis showed a melting / decomposition endotherm (ΔH = 111 J / g) with an onset at 148°C. TGA analysis showed negligible (0.1%) total weight loss up to 145°C, indicating that Form B is insoluble. DVS revealed that the API is non-hygroscopic, exhibiting a weight change of approximately 0.1% at 25°C and 5-95% RH. Samples recovered after dynamic vapor sorption experiments showed no change in solid state morphology by PXRD.

[0206] Kinetic solubility was evaluated in biorelevant media at room temperature at 1, 4, and 24 hours and is summarized in Table 3. Form B Tris salt was virtually insoluble in fasted simulated gastric fluid (FaSSGF) after 1, 4, and 24 hours (0.46, 0.35 μg / mL, and 0.39 μg / mL, respectively). Solubility in fasted simulated intestinal fluid (FaSSIF) was at least 3 mg / mL (parent equivalent) but gradually decreased from 1 hour (1112 μg / mL), 4 hours (851 μg / mL), to 24 hours (733 μg / mL).

[0207] The kinetic solubility in fed simulated intestinal fluid (FeSSIF) was also 3 mg / mL (parent equivalent) in solution, followed by precipitation (99 μg / mL at 1 hour, 96 μg / mL at 4 hours, and 95 μg / mL at 24 hours).

[0208] [Table 4]

[0209] The stability of the solid API was evaluated after 2 weeks of storage under the following conditions: ·25℃ / 58%RH (sealed) ·25℃ / 58%RH (open) ·40℃ / 75%RH (sealed) ·40℃ / 75%RH (open) 80℃ / ambient RH (sealed)

[0210] No significant chemical or physical changes were observed for Tris salt Form B samples stored for 2 weeks at 25°C / 58% RH (closed and open), 40°C / 75% RH (closed and open), and 80°C / ambient RH (closed). PXRD analysis of all stability samples also showed no significant changes. The solid-state stability evaluation of the API is summarized in Table 4.

[0211] [Table 5]

[0212] Pharmacokinetic assays The Tris salt and free acid of Compound 1 were tested in two animal models to evaluate pharmacokinetic parameters.

[0213] Rat model Dosing Groups, Dosing, and Collection. The pharmacokinetics of Compound 1 free acid and Tris salt were evaluated in male Sprague-Dawley rats by oral gavage. Three rats per group were orally administered 20, 60, or 200 mg of Compound 1 free acid, or 25, 75, or 250 mg in 0.5% methylcellulose in saline to give final concentrations of Compound 1 free acid of 2, 6, or 20 mg / mL, or 2.5, 7.5, or 20 mg / mL. Plasma was collected from the jugular vein at 5, 15, and 30 minutes, 1, 2, 4, 6, 8, and 24 hours after administration. No abnormal clinical signs were observed.

[0214] Stock and Dose Preparation. Stock solutions were prepared by dissolving 2.47 mg of Compound 1 (free acid) in 2.470 mL of DMSO with vortexing to give a 1 mg / mL solution of Compound 1 (free acid), or by dissolving 2.09 mg of Compound 1 (Tris salt) in 1.655 mL of DMSO with vortexing to give a 1 mg / mL solution of Compound 1 (Tris salt). Dose solutions were prepared by vortexing / sonicating the following solids in solvent:

[0215] [Table 6]

[0216] LC MS-MS Analysis. Liquid chromatography with tandem mass spectrometry was used to measure plasma concentrations of Compound 1 free acid and Compound 1 Tris salt in plasma samples collected at predetermined time points.

[0217] Standard dilutions of appropriate concentrations were obtained by diluting the analyte stock solution with 50% aqueous acetonitrile. Five μL of the standard dilutions (10, 20, 50, 100, 500, 1000, 5000, 8000, and 10,000 ng / mL) were added to 50 μL of blank male SD rat plasma to achieve calibration standards ranging from 1 to 1,000 ng / mL (1, 2, 5, 10, 50, 100, 500, 800, and 1,000 ng / mL) in a total volume of 55 μL. Five quality control (QC) samples of plasma were prepared independently from those used for the calibration standards: 2 ng / mL, 5 ng / mL, 10 ng / mL, 50 ng / mL, and 800 ng / mL. These QC samples were prepared on the day of analysis, just like the standard dilutions. 55 μL of the standard, 55 μL of the QC sample, and 55 μL of the unknown sample (50 μL of plasma / blood with blank solution) were each added to 200 μL of a methanol mixture containing the internal standard (dexamethasone) for protein precipitation, vortexed for 30 seconds, and centrifuged at 4°C and 4000 rpm for 15 minutes. The supernatant was diluted 1:2 with water. 2 μL of the supernatant was injected into the LC / MS / MS system for quantitative analysis.

[0218] The instrumentation included a HALO 90A C18 2.7 μm 2.1 × 50 mm HPLC column, a Prominence gas unit DGU-20A5R(C), a Shimadzu LC-30AD liquid chromatograph equipped with a communication bus module CBM-20A and an Auto SIL-20AC HT; and an AB Sciex Triple Quad 5500 LC / MS / MS instrument. The following conditions were used:

[0219] [Table 7]

[0220] Results. Results are shown in Table 6 (free acid) and Table 7 (Tris salt), as well as Figures 13-16. Note that the formula weight is higher for the Tris salt (FW = 582.7) than for the free acid (MW = 461.6), so the appropriate comparison is 25 mg / kg Tris salt versus 20 mg / kg free acid. Overall, the total drug exposure (AUC last and AUC Inf ) was higher in the Tris salt group than in the free acid group for both men and women. max ) was also higher in the Tris salt group than in the free acid group in both sexes, except for females given 20 / 25 mg / kg, where the C max (78,883ng / mL) is the C of Tris salt max (68,567 ng / mL). In general, exposure was higher in female than in male subjects.

[0221] [Table 8]

[0222] [Table 9]

[0223] Canine model Groups, Dosing, and Collection. The pharmacokinetics of Compound 1 free acid and Tris salt administered via oral capsules were evaluated in dogs. Three male beagle dogs received a single 3 mg / kg dose of Compound 1 free acid on study day 1 and a single 3 mg / kg dose of Compound 1 Tris salt on study day 8. Plasma was collected by peripheral vein puncture pre-dose and 0.5, 1, 2, 4, 8, 12, and 24 hours post-dose. No abnormal clinical signs were observed.

[0224] In a follow-up study, the pharmacokinetics of the Tris salt Compound 1 was investigated by oral gavage of a 0.5% methylcellulose saline formulation. Two groups of animals were studied. In Group 1, three male beagle dogs received a single dose of 3 mg / kg of the Tris salt of Compound 1. In Group 2, three male beagle dogs received a single dose of 30 mg / kg of the Tris salt of Compound 1. Plasma was collected by peripheral venipuncture before administration and at 0.5, 1, 2, 4, 8, 12, and 24 hours post-dose. No abnormal clinical signs were observed.

[0225] LC MS-MS Analysis. Liquid chromatography with tandem mass spectrometry was used to measure plasma concentrations of Compound 1 free acid and Compound 1 Tris salt in plasma samples collected at predetermined time points. 18 The equipment included a Shimadzu LC-30AD liquid chromatograph equipped with an S-5 μm (50 × 2.1 mm) HPLC column, a CBM-20A communication bus module, and an Auto SIL-20AC HT; and a Triple Quad 5500 LC / MS / MS instrument. Tolbutamide was used as the internal standard. The following conditions were used:

[0226] [Table 10]

[0227] [Table 11]

[0228] Results. Results from the powder capsules are shown in Table 8 and Figures 17 and 18. Results from oral gavage of the suspension are shown in Table 9 and Figures 19 and 20. Overall, total drug exposure (AUC last and AUC Inf) was higher for the Tris salt (both powder capsule and suspension formulations) compared with the free acid. The half-life was shorter for the Tris salt compared with the free acid, but the mean residence time was longer. Because one animal in the salt powder capsule study had abnormally high plasma concentrations at the first two time points (30-50 times higher than the other two subjects), SD and CV% were not calculated.

[0229] [Table 12]

[0230] [Table 13]

[0231] Example 4: Single Crystal A variety of solvent systems (n = 4–8) were selected to grow single crystals of the compounds. The solvents used exhibited a range of polarities, dielectric constants, and dipole moments, and possessed diverse hydrogen bond donor / acceptor properties. The solvents were selected based on their solubility properties (value and temperature dependence) and suitability for single crystal growth experiments.

[0232] Attempts to produce single crystals have included the following crystallization techniques: Two cooling modes with five solvent systems. · Slow evaporation using five solvent systems. · Vapor diffusion using a combination of four solvents. Convective mode with one solvent if the solubility is relatively low and sufficient material is available. Layering using two solvent systems.

[0233] Polarized light microscopy and PXRD were used to assess the crystallinity, size, and quality of the solids obtained from each crystallization experiment. PXRD was used to confirm that the desired Form 2 (Group B) was obtained.

[0234] The single crystal growth study included 24 experiments. Solids were produced in 10 of the 24 experiments. The crystals observed were generally acicular, such as needles or thin blades. Most crystals were deemed too small and thin to be suitable for SCXRD using laboratory-based equipment. One batch of crystals obtained from vapor diffusion (grown from a DMF solution of Tris salt and acetonitrile vapors diffused in the DMF solution over a period of 7 weeks) produced larger crystals and were submitted to a crystallographer for single crystal X-ray diffraction analysis. The results of the crystal growth experiments are summarized below.

[0235] [Table 14]

[0236] [Table 15]

[0237] [Table 16]

[0238] [Table 17]

[0239] [Table 18]

[0240] Single crystals of Form 2 (Group B) grown by vapor diffusion into a solution of the Tris salt in dimethylformamide using acetonitrile as the antisolvent were analyzed by single crystal X-ray diffraction analysis. Powder X-ray diffraction pattern simulations were calculated using Panalytical X'Pert Pro HighScore Plus, v. 2.2.0, using the low-temperature structure and room-temperature unit cell parameters. The simulated and experimental PXRD patterns were based on copper Kα X-rays as the X-ray source.

[0241] [Table 19]

[0242] [Table 20]

[0243] Example 5: Summary of Safety, Toxicity, and Nonclinical Pharmacology and Pharmacokinetics The safety pharmacology of compound 1 and its tris salt was evaluated using an in vitro human ether-a-go-go related gene (hERG) assay, behavioral testing in Sprague-Dawley rats, and respiratory and cardiovascular assessments in electrocardiogram (ECG)-telemetered beagle dogs. Compound 1 exhibited a median inhibitory concentration (IC) of 100% on the hERG channel. 50 ) >300 μM. 50 Measurements of β-amyloid β were excluded due to potential cytotoxicity at doses >300 μM. No Compound 1-related behavioral effects were observed following oral administration of up to 400 mg / kg (highest dose tested) to male rats. No significant respiratory effects were observed following oral administration of up to 150 mg / kg (highest dose tested) to dogs. Cardiovascular safety pharmacological evaluations showed a decrease in PR interval duration, which was detectable up to 9 hours after administration of Compound 1 and was generally dose-related and associated with potential changes in heart rate. These changes were considered biologically insignificant due to their small magnitude (up to 10 ms compared to vehicle control), their association with potential increases in heart rate, and the lack of correlative findings.

[0244] The absorption, distribution, metabolism, and excretion properties of Compound 1 were characterized in both in vitro and in vivo studies. Sensitive and selective bioanalytical methods have been developed and validated for rat and dog plasma. Following intravenous administration, systemic clearance of Compound 1 appeared to be low in all species evaluated. Following oral administration, bioavailability generally exceeded 30% in the species evaluated. Initial studies used the free form of Compound 1. However, due to its higher bioavailability, the Tris salt form of Compound 1 was selected for further development. See Tables 6, 7, and 10. All Good Laboratory Practice (GLP) studies were conducted using the Tris salt form of Compound 1. Dose levels represent the free form equivalent of Compound 1 unless otherwise noted.

[0245] [Table 21]

[0246] Sprague-Dawley rats were administered Compound 1 by oral gavage at doses of 25, 75, and 250 mg / kg / day (females) and 40, 120, and 400 mg / kg / day (males) for 28 days. Compound 1-related deaths occurred in 8 of 15 males treated with 400 mg / kg / day from days 8 to 24. Compound 1-related effects in males treated with 400 mg / kg / day and females treated with 250 mg / kg / day included weight loss, decreased food consumption, clinical pathological changes in hematology and serum chemistry parameters, and histopathological changes. Target tissues included bone marrow, spleen, thymus, liver, lymph nodes, epididymis, seminal vesicles, prostate, testis, uterus, and ovaries. Compound 1 was administered by oral gavage to male rats at 40 or 120 mg / kg / day and to female rats at 25 or 75 mg / kg / day once daily (QD) for 4 weeks, followed by a 4-week recovery period. Compound 1 was well tolerated. Based on these adverse findings, the maximum observed concentration (C max ) were 180,000 ng / mL and 198,000 ng / mL, respectively, and the AUC 0~24The no observed adverse effect levels (NOAELs) were considered to be 120 mg / kg / day in male rats and 75 mg / kg / day in female rats, based on the areas under the concentration-time curve (AUC) over the 24-week period (24 hours) of 2,780,000 ng·h / mL and 3,230,000 ng·h / mL, respectively.

[0247] In a dose-ranging study, Compound 1 was administered by oral gavage to beagle dogs at doses of 30, 79, 240, or 790 mg / kg. Watery stools and vomiting were occasionally observed in dogs at all dose levels. In male dogs administered Compound 1 at 790 mg / kg, potassium increased and sodium and chloride decreased, indicating a possible Compound 1-related effect on electrolyte homeostasis. Decreased thymus weight was observed at ≥240 mg / kg and correlated with microscopic findings of decreased thymic lymphocytes. In a pivotal multiple-dose toxicity study, Compound 1 was administered at doses of 10, 50, and 150 mg / kg / day for 28 days. Based on the lack of adverse findings, the NOAEL in dogs was considered to be 150 mg / kg / day, and the relevant C in both sexes was not observed. max AUC of 80,400 ng / mL in both sexes 0~24 was 474,000 ng·h / mL.

[0248] Example 6: Effect of Compound 1 in combination with approved IPF therapeutic agents in a mouse bleomycin-induced pulmonary fibrosis model (therapeutic treatment) Compound 1 attenuates fibroblast-to-myofibroblast transition and reduces the production of the extracellular matrix protein α-smooth muscle actin (αSMA) in primary human lung fibroblasts. Compound 1 also attenuates pulmonary fibrosis in a bleomycin-induced IPF mouse model when administered at a minimally effective dose of 3 mg / kg twice daily (BID). The reduction in pulmonary fibrosis upon MCT4 inhibition was accompanied by changes in serum metabolites indicating increased extracellular matrix protein turnover and fatty acid oxidation. Compound 1 also attenuated pulmonary fibrosis, along with reduced lung lactate levels, in a bleomycin-induced IPF mouse model in aged (>60 weeks) mice.

[0249] The effects of Compound 1 alone or in combination with pirfenidone or nintedanib, approved IPF treatments, were tested in a bleomycin-induced pulmonary fibrosis model in a therapeutic mode. Compound 1 was administered at 3 mg / kg twice daily; pirfenidone at 100 mg / kg twice daily; and nintedanib at 50 mg / kg once daily, all administered by oral gavage. The results of this study are shown in Figure 23.

[0250] The addition of pirfenidone or nintedanib to Compound 1 did not result in a statistically significant additive effect on either Ashcroft score or αSMA quantification.

[0251] Example 7: Phase I Protocol Phase 1 studies will be randomized, double-blind, placebo-controlled, single- and multiple-ascending dose studies, with sequential arms in each study part. All Phase 1 studies will be conducted using Compound 1 Tris salt unless otherwise indicated. The objectives and endpoints of this study are shown in Table 11.

[0252] [Table 22]

[0253] [Table 23]

[0254] Part A will evaluate the safety, tolerability, and pharmacokinetics (PK) and food effect of a single dose of Compound 1 Tris salt in healthy subjects, and Part B will evaluate multiple oral doses in healthy subjects. Part C will evaluate the safety, tolerability, and PK of a single dose of Compound 1 Tris salt in healthy elderly subjects. Part D is an exploratory, multiple-dose, placebo-controlled, safety, tolerability, PK, and pharmacodynamic (PD) study conducted in patients with idiopathic pulmonary fibrosis (IPF). Parts A, B, and C will be conducted at a single center, and Part D will be conducted at multiple clinical centers.

[0255] Doses may be adjusted downward or upward, delayed, not administered, or repeated. Dose escalation between arms in Parts A and B will be no more than 5-fold for anticipated non-pharmacologically active dose levels and no more than 3-fold for anticipated pharmacologically active dose levels. The interval between dose escalations will be a minimum of 6 days to allow sufficient time for adequate safety review. Schematic diagrams of the planned arms for Parts A and C, and Parts B and D, assuming dose level escalation for each subsequent arm and sentinel administration for each single ascending dose (SAD) arm in Part A, are shown in Figures 21 and 22, respectively.

[0256] Part A: Single ascending dose Part A will include a single-dose, sequential group design. Five treatment arms will be designed in which healthy subjects will be randomly assigned to receive a single oral dose of Compound 1 Tris salt or placebo, with one arm including a two-period crossover arm to examine the effect of food. Sentinel dosing will be used in all SAD arms. The SAD portion will include: Screening visit 2 days to 4 weeks before administration of the investigational drug (IMP). The clinical treatment period will be 4 days, with check-in to the study site on Day -1 (including 48 hours post-dose). On Day 1, eligible subjects will be randomized to Compound 1 Tris salt or placebo prior to dosing. A safety follow-up will be conducted 10 (±1) days after dose administration.

[0257] Dosing will occur on Day 1. Safety assessments will include adverse events (AEs), clinical laboratory tests (hematology, clinical chemistry, urinalysis), electrocardiogram (ECG), vital signs, and physical examination. Blood and urine samples to determine the PK profile of Compound 1 Tris salt will be collected from pre-dose through 48 hours post-dose.

[0258] Part B: Multiple Ascending Dose Part B involves a multiple-dose, sequential group study. Three dose groups of healthy subjects are planned to be randomized to multiple oral doses of Compound 1 Tris salt or placebo. The apparent terminal elimination half-life (t 1 / 2If the duration of treatment with steroids (SDS) in Part A is found to be shorter or longer than predicted from nonclinical data, further dose escalation may be necessary to include modified dosing regimens for Parts B and D. The multiple dose escalation portion includes: Screening visit between 2 days and 4 weeks before the first IMP dose. The clinical treatment period will be 10 days, with check-in to the study site on Day -1, including 48 hours after the last dose. Eligible subjects will be randomized to Compound 1 Tris salt or placebo on Day 1 prior to dosing. A safety follow-up visit will be conducted 10 (±2) days after the last dose.

[0259] All subjects will be scheduled to receive dosing once daily (QD) at approximately the same time each morning on Days 1 through 7 (inclusive). The total daily dose administered will not exceed the exposure demonstrated to be safe and well-tolerated. Safety assessments will include AEs, clinical laboratory tests (hematology, clinical chemistry, and urinalysis), ECG, vital signs, and physical examination. Blood samples will be collected prior to the first dose on Day 1 and 4 hours post-dose on the final dose for analysis of 3-methylhistidine and a metabolomics panel. Blood samples for determination of the PK profile of Compound 1 Tris salt will be collected from pre-dose through 24 hours post-dose on Day 1; pre-dose on Days 4 through 6; and from pre-dose through 48 hours post-dose on Day 7.

[0260] Part C: Single dose in healthy elderly subjects Part C involves a single-dose, single-arm, randomized design to evaluate safety, tolerability, and PK in healthy elderly subjects to ensure safety in elderly subjects prior to administration to patients with IPF. One group of healthy elderly subjects will be randomized to receive a single oral dose of Compound 1 Tris salt or placebo. Single-dose evaluations will include the following: Screening visit between 2 days and 4 weeks before the first IMP dose. The clinical treatment period will be 4 days, including 48 hours after the last dose, with check-in to the study site on Day -1. Eligible subjects will be randomized to Compound 1 Tris salt or placebo on Day 1 prior to dosing. A safety follow-up visit will be conducted 10 (±1) days after the last dose.

[0261] Dosing will occur on Day 1. Safety assessments will include AEs, clinical laboratory tests (hematology, clinical chemistry, and urinalysis), electrocardiogram, vital signs, and physical examination. Blood and urine samples to determine the PK profile of Compound 1 Tris salt will be collected from pre-dose through 48 hours post-dose.

[0262] Part D: Multiple doses in patients with idiopathic pulmonary fibrosis Part D will include a multiple-dose, single-arm, randomized design to evaluate safety, tolerability, PK, and PD in patients with IPF. The patient portion of the study will include: Screening visit between 2 days and 5 weeks before the first IMP dose. A minimum of 7 days and a maximum of 28 days of outpatient treatment (including a Day 1 outpatient visit and subsequent weekly outpatient visits). Eligible patients will be randomized to Compound 1 Tris salt or placebo on Day 1 prior to dosing. A safety follow-up survey will be conducted 10 (±2) days after the last dose.

[0263] For all patients, dosing is planned QD at approximately the same time each morning. Dosing will occur at the study site on Day 1 and other outpatient visits, with patients self-administering on the remaining days. The dose level selected for Part D will be at or below the level evaluated as safe and well-tolerated in Part B. Safety assessments will include AEs, clinical laboratory values (hematology, clinical chemistry, and urinalysis), electrocardiogram, vital signs, and physical examination. Blood samples for PK analysis will be collected on Day 1 and the last day of dosing at time points determined based on the PK analysis results in Parts A, B, and C, and before dosing on other outpatient visits.

[0264] Dynamic positron emission tomography of the lung using the radioactive tracer 2[fluorine-18]-fluoro-2-deoxy-D-glucose will be performed at screening and weekly to assess pulmonary glucose uptake. Blood samples will be collected for 3-methylhistidine analysis and a metabolomics panel before the first dose on Day 1 and 4 hours after the final dose. Pulse oximetry to assess oxygen saturation (SpO2); spirometry to assess forced vital capacity (FVC), forced expiratory volume in seconds (FEV1), and FEV1 / FVC; and diffusing capacity for carbon monoxide (DLCO) testing will be performed at screening and at each outpatient visit.

[0265] Number of subjects Each group in Parts A, B, and C will have 8 subjects randomized 6:2 to Compound 1 Tris salt:placebo. In Part A, the total number of subjects will be approximately 40 (8 subjects x 5 groups). In Part B, the total number of subjects will be approximately 24 (8 subjects x 3 groups). In Part C, the total number of subjects will be approximately 8 (8 subjects x 1 group). Part D will involve at least 8 (and possibly up to 16) IPF patients in each arm, with a 3:1 ratio of active agent to placebo.

[0266] Additional groups may be used depending on the need for more evaluation, and groups may be removed based on the data obtained.

[0267] Diagnosis and main inclusion criteria Parts A and B: Body mass index at screening of 18.0 to 32.0 kg / m 2 Healthy male and female subjects aged 18-60 years (inclusive).

[0268] Part C: Body mass index at screening of 18.0–32.0 kg / m 2 Healthy male and female subjects aged 65-80 years (inclusive).

[0269] Group D: Male and female patients aged 40 to 80 years (inclusive) who were diagnosed with IPF based on a multidisciplinary team review and high-resolution computed tomography performed within one year prior to informed consent, in accordance with the 2018 American Thoracic Society / European Respiratory Society / Japanese Respiratory Society / Latin American Thoracic Society Clinical Practice Guidelines for the Diagnosis of IPF. Patients were required to meet the following criteria: SpO2 ≥ 90% at rest by pulse oximetry while breathing ambient air, FVC and FEV1 ≥ 50% of predicted, FEV1 to FVC ratio ≥ 0.7, and DLB after adjustment for hemoglobin. CO must be between 30% and 79% (inclusive) of the predicted value.

[0270] Exclusion criteria Parts A, B, and C: Parts A, B, and C subjects will be excluded from the study if they meet any of the following criteria at the screening visit, unless otherwise indicated or approved by the investigator (or designee): ·Significant medical history or clinical symptoms of metabolic, allergic, skin, hepatic, renal, hematological, pulmonary, cardiovascular, gastrointestinal, neurological, respiratory, endocrine, or psychiatric disorders; significant medical history of hypersensitivity, intolerance, or allergy to any drug compound, food, or other substance; and / or history of stomach or intestinal surgery or resection that may alter the absorption and / or excretion of orally administered drugs (uncomplicated appendectomy and hernia repair are permitted; cholecystectomy is not permitted). A single 12-lead ECG at screening shows any of the following: QTcF >450 ms (men) or >470 ms (women), confirmed by calculation of the original value and the mean of two replicates. QQRS duration >110ms, confirmed by calculation of the original value and the average of two repetitions. PR interval >220 ms, confirmed by calculating the original value and the mean of two repetitions. Findings that make it difficult to measure the QT interval corrected for heart rate (QTc) or that make the QTc data uninterpretable. - History of additional risk factors for torsades de pointes (e.g., heart failure, hypokalemia, family history of long QT syndrome). Pulse rate ≥ 100 beats / min or < 40 beats / min at screening. -Positive hepatitis panel test and / or positive human immunodeficiency virus test. · Administration of a COVID 19 vaccine within the past 30 days prior to administration. - have used or plan to use any medication / product known to alter drug absorption, metabolism, or excretion processes, including St. John's wort, within 30 days prior to dosing; have used or plan to use any prescription medication / product within 14 days prior to dosing; have used or plan to use any sustained-release medication / product that is believed to still be active within 14 days prior to check-in; and / or have used or plan to use any over-the-counter medication / product, including vitamins, minerals, and botanicals / herbs / plant-derived preparations, within 7 days prior to check-in. Participation in a clinical trial involving administration of an investigational new chemical entity within the past 30 days or 5 half-lives (whichever is longer) prior to administration. - Have previously completed or discontinued a study testing Compound 1, or have previously received Compound 1. Alcohol intake of >21 units per week for men and >14 units per week for women. · A positive urine drug test at screening or check-in, or a positive alcohol test at check-in. · History of alcoholism or drug / chemical abuse within two years prior to check-in. Use of tobacco or nicotine-containing products within three months prior to check-in, or a positive cotinine test at screening or check-in. Consumption of poppy seeds, Seville oranges, or grapefruit-containing foods or drinks within seven days prior to check-in. -Administration of blood products within two months prior to check-in. Donation of blood within 3 months prior to screening, plasma within 2 weeks prior to screening, or platelets within 6 weeks prior to screening. ·Poor peripheral venous access. Parts A and B: Systolic blood pressure >140mmHg or <90mmHg, or diastolic blood pressure >90mmHg or <50mmHg at screening and check-in. Small deviations from this range may be permitted if deemed by the investigator to be not clinically significant. Part C: Systolic blood pressure >150mmHg or <90mmHg, or diastolic blood pressure >100mmHg or <50mmHg at screening and check-in. Small deviations from this range may be permitted if not deemed clinically significant by the investigator.

[0271] Part D: Part D patients will be excluded from the study if they meet any of the following criteria at the screening visit, unless otherwise indicated or approved by the investigator (or designee): Serious or uncontrolled medical, surgical, or psychiatric illness that, in the opinion of the investigator, may compromise patient safety. History of acute exacerbation of IPF within 3 months prior to screening, history of malignancy (excluding treated squamous cell carcinoma and basal cell carcinoma and treated stage 0 / non-invasive cervical cancer) within 5 years prior to screening, and / or history of emphysema or clinically significant respiratory disease (other than IPF). Surgery scheduled during the study period (from Day 1 through the follow-up visit). Findings on surgical lung biopsy (previous), HRCT imaging, transbronchial lung biopsy (previous), or bronchoalveolar lavage (previous) that are diagnostic of another disease other than UIP. Other known causes of interstitial lung disease (e.g., drug toxicity, environmental exposure, connective tissue disease). End-stage fibrotic disease expected to require an organ transplant within six months. - Clinically significant findings from medical history (other than IPF), 12-lead ECG, vital signs, or laboratory tests that may compromise patient safety. -Positive hepatitis panel test and / or positive human immunodeficiency virus test. - Killed and inactivated vaccines (e.g., pneumonia, influenza) up to ≤14 days prior to screening, or live attenuated vaccines (e.g., chickenpox) up to ≤2 months prior to screening. Administration of systemic corticosteroids, cytotoxic therapy (e.g., chlorambucil, azathioprine, cyclophosphamide, or methotrexate), vasodilator therapy for pulmonary hypertension (e.g., bosentan), or unapproved treatments for IPF (e.g., interferon-gamma, penicillamine, cyclosporine, mycophenolate, N-acetylcysteine) within 4 weeks prior to screening. Treatment with pirfenidone or nintedanib (but not both drugs simultaneously) is permitted only if the patient has been on a stable dose for at least 4 weeks prior to screening and is expected to remain on that dose throughout the entire enrollment period (i.e., from signing the ICF to the last protocol-specified assessment, whether scheduled or unscheduled). - have used or plan to use any medication / product known to alter drug absorption, metabolism, or excretion processes, including St. John's wort, within 30 days prior to dosing; have used or plan to use any prescription medication / product within 14 days prior to dosing; have used or plan to use any sustained-release medication / product that is believed to still be active within 14 days prior to check-in; and / or have used or plan to use any over-the-counter medication / product, including vitamins, minerals, and botanicals / herbs / plant-derived preparations, within 7 days prior to dosing on Day 1. Participation in a clinical trial involving administration of an investigational new chemical entity within the past 30 days or 5 half-lives (whichever is longer) prior to the Day 1 dose. - Have previously completed or discontinued a study testing Compound 1, or have previously received Compound 1. Alcohol consumption >21 units per week for men and >14 units per week for women. One unit of alcohol is: beer: 12 oz (360 mL), hard liquor: 1 1 / 2 oz (45 mL), or wine: 5 oz (150 mL). - A positive screening or Day 1 urine drug (including cotinine) test result or a positive Day 1 alcohol test result. History of alcoholism or drug / chemical abuse within 2 years prior to Day 1 dose. Use of tobacco or nicotine-containing products within 3 months prior to Day 1 dose. - Consumption of poppy seeds, Seville oranges, or grapefruit-containing foods or beverages within 7 days prior to the Day 1 dose. -Administration of blood products within 2 months prior to Day 1 dose administration. Donation of blood from 3 months prior to Day 1 dose, plasma from 2 weeks prior to screening, or platelets from 6 weeks prior to screening. ·Poor peripheral venous access.

[0272] Dose selection Compound 1 Tris salt was evaluated for safety in 7-day and 28-day studies in rats and dogs. Compound 1 Tris salt-related effects in a 28-day rat GLP study were weight loss, decreased food consumption, clinical pathological changes in hematology and serum chemistry parameters, and histopathological changes at 400 mg / kg / day in males and 250 mg / kg / day in females. The NOAEL in this rat study was 120 mg / kg / day in males and 75 mg / kg / day in females. No adverse effects related to Compound 1 Tris salt were observed in a 28-day dog GLP study. The NOAEL determined in the 28-day dog GLP study was 150 mg / kg / day, which corresponds to the following human equivalent dose (HED): Rat: 75 mg / kg x 0.16 = 12 mg / kg Dogs: 150 mg / kg x 0.54 = 81 mg / kg

[0273] where 0.16 and 0.54 are conversion factors for extrapolating animal dose to HED based on body surface area.

[0274] Rats are the most sensitive species (i.e., the species with the lowest HED), and assuming a 10-fold safety margin, this corresponds to the following maximum recommended starting dose:

number

[0275] Using the principles of allometry, human PK parameters were predicted based on the single oral dose PK parameters. Based on the predicted human PK parameters and the minimal effective dose in mouse IPF studies, the clinically effective dose was predicted to be approximately 30-80 mg.

[0276] Investigational drug, dosage, and administration method 25 mg and 100 mg Compound 1 Tris Salt Capsules. Route of Administration: Oral.

[0277] Part A dosing plan: A single starting dose of 50 mg, followed by planned doses of 75, 100, 150, and 200 mg of Compound 1 Tris salt. Dose escalation between groups will be no more than three-fold. Dose escalation will be performed only when data from at least six subjects from the previous lower dose group have been reviewed, ensuring that data from at least four subjects receiving Compound 1 Tris salt are used to make dose escalation decisions.

[0278] Proposed Dosing for Part B: The dose, frequency, and duration of administration for Part B will be determined in consultation with the sponsor based on the data from Part A of the study. The total daily exposure of Compound 1 Tris salt administered during this part of the study will not exceed the level demonstrated to be safe and well tolerated in Part A. Dose escalation will be performed only after data from at least six subjects from the previous lower dose cohort have been reviewed, ensuring that data from at least four subjects receiving Compound 1 Tris salt are used to make dose escalation decisions. Dose escalation between cohorts will be no more than three-fold.

[0279] Proposed Dosing for Part C: The dosing for Part C will be determined in consultation with the sponsor based on the data from Part A of the study. Healthy elderly subjects will be dosed at a level not exceeding the maximum dose found to be safe and well-tolerated in Part A.

[0280] Proposed Dosage for Part D: The dose, frequency, and duration of administration for Part D will be determined in consultation with the sponsor based on the preliminary data from Part B of the study. The total daily exposure of Compound 1 Tris salt administered in Part D will not exceed the exposure demonstrated to be safe and well tolerated in Part B. Part D will not commence until review of safety and tolerability data from single-dose administration to healthy elderly subjects in Part C.

[0281] The food state for groups in Part A prior to the food effect group will be fasted. Based on review of preliminary PK data from the food effect group, the food state for subsequent groups in Part A may be either fasted or fed. The food state for dosing in Parts B, C, and D will be determined after review of preliminary PK data (if applicable) from the food effect group in Part A and earlier groups in Part B.

[0282] All references, patents, or applications (U.S. or foreign) cited in this application are hereby incorporated by reference as if set forth in their entirety herein. In the event of a conflict, the present disclosure will control literally.

[0283] From the foregoing description, those skilled in the art can easily ascertain the essential features of the present disclosure, and can make various changes and modifications to the present disclosure to adapt it to various applications and conditions without departing from the spirit and scope thereof.

Claims

1. Structural formula I 【Chemical 1】 Form A of the compound of formula (I).

2. The compound of claim 1 which is unsolvated.

3. 3. The compound of claim 1 or 2, wherein the compound has differential scanning calorimetry data showing a first-order melting endotherm with an onset of about 157°C.

4. 4. The compound of claim 3, wherein the compound has a differential scanning calorimetry trace substantially as shown in FIG.

5. 5. The compound of any one of claims 1 to 4, wherein the compound has a TGA trace substantially as shown in Figure 2.

6. The compound according to any one of claims 1 to 5, having an FT-Raman spectrum substantially as shown in Figure 1.

7. 7. The compound of claim 6, having an X-ray powder diffraction (XRPD) pattern substantially as shown in FIG.

8. Amorphous structural formula I 【Chemistry 2】 Compound.

9. Structural formula II 【Chemistry 3】 Form A of the compound of formula (I).

10. 10. The compound of claim 9, which is unsolvated.

11. 11. The compound of claim 9 or 10, wherein the compound has differential scanning calorimetry data showing a melting endotherm with an onset of about 147°C.

12. 12. The compound of claim 11, wherein the compound has a differential scanning calorimetry trace substantially as shown in Figure 6.

13. 13. The compound of any one of claims 9 to 12, wherein the compound has a TGA trace substantially as shown in Figure 6.

14. Approximately 1604, 1438, and 995 cm －１ The compound according to any one of claims 9 to 13, characterized by the presence of an FT-Raman peak of

15. Approximately 1604, 1438, 1372, 995, 332, 234, and 173 cm －１ The compound of claim 14 characterized by an FT-Raman peak of:

16. 15. The compound of claim 14, having an FT-Raman spectrum substantially as shown in FIG.

17. 17. The compound of any one of claims 9-16, having an X-ray powder diffraction (XRPD) pattern with peaks at about 7.79, 15.61, 16.71, 20.00, and 20.88±0.3 degrees two-theta, wherein the XRPD is measured using an incident beam of Cu radiation.

18. 18. The compound of claim 17, having an X-ray powder diffraction (XRPD) pattern with peaks at about 7.79, 12.59, 15.61, 16.71, 20.00, 20.88, and 21.50±0.3 degrees 2θ, wherein the XRPD is measured using an incident beam of Cu radiation.

19. 20. The compound of claim 17, having an X-ray powder diffraction (XRPD) pattern with peaks at about 7.79, 12.18, 12.59, 15.61, 16.71, 17.38, 17.72, 19.16, 20.00, 20.88, and 21.50±0.3 degrees 2θ, wherein the XRPD is measured using an incident beam of Cu radiation.

20. 18. The compound of claim 17, having an X-ray powder diffraction (XRPD) pattern with peaks at d-spacings of about 11.34, 5.67, 5.30, 4.44, and 4.25±0.3 Å, wherein the XRPD is measured using an incident beam of Cu radiation.

21. 20. The compound of claim 17, having an X-ray powder diffraction (XRPD) pattern with peaks at about 11.34, 7.02, 5.67, 5.30, 4.44, 4.25, and 4.13±0.3 Å d-spacing, wherein the XRPD is measured using an incident beam of Cu radiation.

22. 20. The compound of claim 17, having an X-ray powder diffraction (XRPD) pattern with peaks at about 11.34, 7.26, 7.02, 5.67, 5.30, 5.10, 5.00, 4.63, 4.44, 4.25, and 4.13 Å in d-spacing, wherein the XRPD is measured using an incident beam of Cu radiation.

23. 18. The compound of claim 17, having an X-ray powder diffraction (XRPD) pattern substantially as shown in Figure 7.

24. 1. A process for preparing Form A of the compound of structural formula II, comprising combining a compound of structural formula I with tris(hydroxymethyl)aminomethane in a solvent, and isolating Form A of the compound of structural formula II.

25. Structural formula II 【Chemistry 4】 Form B of the compound.

26. 26. The compound of claim 25, which is unsolvated.

27. 27. The compound of claim 25 or 26, wherein the compound has differential scanning calorimetry data showing a melting endotherm with an onset at about 148°C.

28. 28. The compound of claim 27, wherein the compound has a differential scanning calorimetry trace substantially as shown in Figure 10.

29. 29. The compound of any one of claims 25 to 28, wherein the compound has a TGA trace substantially as shown in Figure 10.

30. Approximately 1601, 1545, 1468, 1437, 999, 995, and 234 cm －１ The compound according to any one of claims 25 to 29, characterized by the presence of an FT-Raman peak of

31. Approximately 2946, 1601, 1545, 1507, 1468, 1437, 1374, 1345, 1043, 999, 995, 284, 234, and 186 cm －１ The compound of claim 30, characterized by the presence of an FT-Raman peak of

32. 32. The compound of any one of claims 25-31, having an X-ray powder diffraction (XRPD) pattern with peaks at about 9.29, 9.70, 16.36, 19.12, and 20.15 degrees two-theta, wherein the XRPD is measured using an incident beam of Cu radiation.

33. 33. The compound of claim 32, having an X-ray powder diffraction (XRPD) pattern with peaks at about 9.29, 9.70, 10.03, 16.36, 19.12, 19.49, 19.61, 20.15, and 21.68 degrees 2θ, wherein the XRPD is measured using an incident beam of Cu radiation.

34. 33. The compound of claim 32, having an X-ray powder diffraction (XRPD) pattern with peaks at about 9.29, 9.70, 10.03, 11.14, 11.73, 16.36, 16.71, 19.12, 19.49, 19.61, 20.15, 20.52, 20.73, and 21.68 degrees 2θ, wherein the XRPD is measured using an incident beam of Cu radiation.

35. 33. The compound of claim 32, having an X-ray powder diffraction (XRPD) pattern with peaks at d-spacings of about 9.51, 9.11, 5.41, 4.64, and 4.40 Å, wherein the XRPD is measured using an incident beam of Cu radiation.

36. 33. The compound of claim 32, having an X-ray powder diffraction (XRPD) pattern with peaks at d-spacings of about 9.51, 9.11, 8.81, 5.41, 4.64, 4.55, 4.52, 4.40, and 4.10 Å, wherein the XRPD is measured using an incident beam of Cu radiation.

37. 33. The compound of claim 32, having an X-ray powder diffraction (XRPD) pattern with peaks at about 9.51, 9.11, 8.81, 7.93, 7.54, 5.41, 5.30, 4.64, 4.55, 4.52, 4.40, 4.32, 4.28, and 4.10 Å d-spacing, wherein the XRPD is measured using an incident beam of Cu radiation.

38. 33. The compound of claim 32, having an X-ray powder diffraction (XRPD) pattern substantially as shown in Figure 11.

39. The unit cell lengths of the three axes are approximately (a) 26.526A, (b) 5.940A, and (c) 19.055A, and the unit cell angles are approximately (a) 90.00°, (β) 90.00°, and (γ) 93.123°. １ 33. The compound of claim 32 characterized by the / c space group.

40. 40. The compound of any one of claims 25 to 39, wherein the compound is stable at 25°C and 58% relative humidity for at least 2 weeks.

41. 40. The compound of any one of claims 25 to 39, wherein the compound is stable at 40°C and 75% relative humidity for at least 2 weeks.

42. 40. The compound of any one of claims 25 to 39, wherein the compound is stable at 80°C and ambient relative humidity for at least 2 weeks.

43. 1. A process for preparing Form B of the compound of structural formula II, comprising stirring Form A of the compound of structural formula II with a suitable solvent, adding seed crystals of Form B of the compound of structural formula II, and isolating Form B of the compound of structural formula II.

44. A pharmaceutical composition comprising a compound according to any one of claims 1 to 42 and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

45. 43. A pharmaceutical composition comprising a compound of any one of claims 1 to 42 and a pharmaceutically acceptable carrier, adjuvant, or vehicle, wherein said composition does not contain detectable Group A Tris salts.

46. 43. A method for inhibiting the activity of the monocarboxylate transporter MCT4, or a mutant thereof, in a biological sample, the method comprising contacting the biological sample with a compound according to any one of claims 1 to 42.

47. 43. A method of inhibiting the activity of the monocarboxylate transporter MCT4, or a mutant thereof, in a patient, comprising administering to said patient a compound according to any one of claims 1 to 42.

48. 43. A method of selectively inhibiting the activity of the monocarboxylate transporter MCT4, or a variant thereof, over the monocarboxylate transporter MCT1, or a variant thereof, in a patient, comprising administering to said patient a compound according to any one of claims 1 to 42.

49. 49. The method of claim 48, wherein the inhibition is at least 100-fold selective for MCT4 over MCT1.

50. 43. A method of treating a monocarboxylic acid transporter MCT4-mediated disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 42.

51. 51. The method of claim 50, wherein the subject is a human.

52. 51. The method of claim 50, wherein the subject is in a fed state.

53. 51. The method of claim 50, wherein the subject is in a fasted state.

54. 51. The method of claim 50, wherein the monocarboxylate transporter MCT4-mediated disorder is selected from an inflammatory disorder and a proliferative disorder.

55. 51. The method of claim 50, wherein the monocarboxylate transporter MCT4-mediated disorder is a proliferative disorder.

56. 56. The method of claim 55, wherein the proliferative disorder is cancer.

57. The cancer is selected from the group consisting of adenocarcinoma, adult T-cell leukemia / lymphoma, bladder cancer, blastoma, bone cancer, breast cancer, brain tumor, carcinoma, myeloid sarcoma, cervical cancer, colon cancer, esophageal cancer, digestive cancer, glioblastoma multiforme, glioma, gallbladder cancer, gastric cancer, head and neck cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma, intestinal cancer, kidney cancer, laryngeal cancer, leukemia, lung cancer, lymphoma, liver cancer, small cell lung cancer, non-small cell lung cancer, mesothelial cancer, and the like.

57. The method of claim 56, wherein the cancer is selected from tumors, multiple myeloma, eye cancer, optic nerve tumor, oral cancer, ovarian cancer, pituitary tumor, primary central nervous system lymphoma, prostate cancer, pancreatic cancer, pharyngeal cancer, renal cell carcinoma, rectal cancer, sarcoma, skin cancer, spinal cord tumor, small intestine cancer, gastric cancer, T-cell lymphoma, testicular cancer, thyroid cancer, pharyngeal cancer, genitourinary cancer, urothelial carcinoma, uterine cancer, vaginal cancer, and Wilms' tumor.

58. 55. The method of claim 54, wherein the monocarboxylate transporter MCT4-mediated disorder is an inflammatory disorder.

59. 59. The method of claim 58, wherein the inflammatory disorder is selected from Crohn's disease, ulcerative colitis, idiopathic pulmonary fibrosis, muscular dystrophy, rheumatoid arthritis, and systemic sclerosis (scleroderma).

60. 60. The method of claim 59, wherein the inflammatory disorder is idiopathic pulmonary fibrosis.

61. 61. The method of any one of claims 50 to 60, wherein the therapeutically effective amount is from about 30 mg to about 200 mg.

62. 62. The method of claim 61, wherein the therapeutically effective amount is from about 30 mg to about 80 mg.

63. 62. The method of claim 61, wherein the therapeutically effective amount is selected from 50 mg, 75 mg, 100 mg, 150 mg, and 200 mg.

64. 43. A method of treating a monocarboxylate transporter MCT4-mediated disorder in a subject in need thereof, comprising the sequential or co-administration of a compound according to any one of claims 1 to 42 and another therapeutic agent.

65. 65. The method of claim 64, wherein the monocarboxylate transporter MCT4-mediated disorder is a metabolic disease.

66. 66. The method of claim 65, wherein the metabolic disease is selected from metabolic syndrome, diabetes, dyslipidemia, fatty liver disease, non-alcoholic steatohepatitis, obesity, and insulin resistance.

67. 67. The method of claim 66, wherein the diabetes is type II diabetes.

68. 67. The method of claim 66, wherein the dyslipidemia is hyperlipidemia.

69. 65. The method of claim 64, wherein the therapeutic agent is selected from paracetamol, acetaminophen, pirfenidone, nintedanib, and non-hormonal contraceptives.

70. 43. A method for achieving an effect in a patient, comprising administering to the patient a therapeutically effective amount of a compound of any one of claims 1 to 42, wherein said effect is selected from the group consisting of reduced triglycerides, reduced cholesterol, and reduced hemoglobin A1c.

71. 71. The method of claim 70, wherein the cholesterol is selected from LDL cholesterol and VLDL cholesterol.

72. 71. The method of claim 70, wherein the triglycerides are selected from plasma triglycerides and liver triglycerides.

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